Griseofulvin_HNMR_13495_MedChemExpress
QuEChERS-超高效液相色谱-三重四极杆串联质谱法测定水果制品和肉酱中10种四环素类抗生素
谷悦，唐会鑫，李朔，等. QuEChERS-超高效液相色谱-三重四极杆串联质谱法测定水果制品和肉酱中10种四环素类抗生素[J].食品工业科技，2023，44（18）：313−320. doi: 10.13386/j.issn1002-0306.2022100072GU Yue, TANG Huixin, LI Shuo, et al. Determination of 10 Tetracycline Antibiotics in Fruit Products and Meat Sauce by QuEChERS with Ultra Performance Liquid Chromatography-Triple Quadrupole Tandem Mass Spectrometry[J]. Science and Technology of Food Industry, 2023, 44(18): 313−320. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2022100072· 分析检测 ·QuEChERS-超高效液相色谱-三重四极杆串联质谱法测定水果制品和肉酱中10种四环素类抗生素谷 悦1，唐会鑫1，李 朔2，马 玲2，王 可1,2, *，杨莉丽1（1.河北师范大学化学与材料科学学院，河北石家庄 050024；2.石家庄市疾病预防控制中心，石家庄市化学毒物检测及风险预警技术创新中心，河北石家庄 050011）摘 要：使用QuEChERS 结合超高效液相色谱-三重四极杆串联质谱（Ultra performance liquid chromatography-triple quadrupole tandem mass spectrometry ，UPLC-MS/MS ），建立检测水果制品和肉酱中10种四环素类抗生素的分析方法。
茶多酚对人脂肪来源间充质干细胞成骨分化的影响
茶多酚对人脂肪来源间充质干细胞成骨分化的影响王华1，齐玉成-杨云芳-赵艺洋2,王慧1，陈培1，杨旭芳1(1.牡丹江医学院，黑龙江牡丹江157011；2.南方医科大学第一临床医学院，广东广州510515)摘要：目的探讨茶多酚(Epigallocatechin-3-gallate,EGCG)对人脂肪间充质干细胞(human adipose-derived mesenchy^-mal stem cells,hADSCs)成骨分化的影响。
方法利用胶原酶消化法和贴壁筛选法从人脂肪组织中分离、培养及扩增hADSCs,倒置显微镜下观察各代hADSCs的形态学特点;利用流式细胞术检测各代hADSCs免疫学表型;取P3代细胞进行成骨诱导分化，实验分三组，即未诱导组、常规成骨诱导组与EGCG组(常规成骨诱导+5^mol/L EGCG),14d后，镜下观察细胞形态学改变及碱性磷酸酶(ALP)染色。
结果体外分离、培养的hADSCs形态均一;流式细胞术结果显示hADSCs具备间充质干细胞的免疫学表型，即CD44、CD73、CD105阳性;成骨诱导14d后部分细胞由长梭形变成多角形，细胞呈现聚集趋势；ALP染色显示EGCG组呈强阳性。
结论成功的从脂肪组织中分离培养出了hADSCs,EGCG能加强其成骨分化能力，这将为骨质疏松症的临床药物开发提供新的思路，亦为组织工程骨的构建提供丰富可靠的种子细胞来源。
关键词：EGCG；人脂肪来源间充质干细胞;成骨分化中图分类号：R595.2文献标识码：A文章编号：1001-7550(2021)01-0001-04Effect of EGCG on osteogenic differentiation of human adipose-derived mesenchymal stem cellsWANG Hua et al(Mudanjiang Medical University,Mudanjiang157011,China)Abstract:Objective To explore the effect of tea polyphenol EGCG on the osteogenic differentiation of human adipose-derived mesenchymal stem cells(hADSCs) .Methods To isolate,culture and amplify hADSCs from human adipose tissue by collagenase digestion and adherent screening methods, the morphological characteristics of each passage of hADSCs were observed under an inverted microscope.The immunophenotype of each generation of hADSCs was detected by flow-cytometry.P3passage cells were taken for osteogenic induction and differentiation,and were divided into three groups:non-induced group, conventional osteogenic induction group and EGCG group(conventional osteogenic induction with+5Rmol/L EGCG).After14days,morphological changes and alkaline phosphatase(ALP)staining were observed under the microscope.Results The morphology of hADSCs isolated and cultured in vitro was uni-form.The results of flow cytometry showed that hADSCs had the immunophenotype of mesenchymal stem cells,such as CD44,CD73and CD105.After14days of osteogenic induction,some cells changed from long spindle shape to polygonal shape,and the cells showed aggregation trend.ALP staining showed strong positive in EGCG group.Conclusion hADSCs have been successfully isolated and cultured from adipose tissue.EGCG can enhance the osteogenic differentiation ability of hADSCs,which will provide a new idea for the clinical drug development of osteoporosis and provide an abundant and reliable source of seed cells for the construction of tissue-engineered bone.Key words:EGCG;human adipose-derived mesenchymal stem cells；osteogenic differentiation随着人口老龄化,骨质疏松症已成为影响人们生活质量的主要因素之一⑷。
marked manuscript
Quality evaluation of Flos Lonicerae through a simultaneous determination of seven saponins by HPLC with ELSDXing-Yun Chai1, Song-Lin Li2, Ping Li1*1Key Laboratory of Modern Chinese Medicines and Department of Pharmacognosy, China Pharmaceutical University, Nanjing, 210009, People’s Republic of China2Institute of Nanjing Military Command for Drug Control, Nanjing, 210002, People’s Republic of China*Corresponding author: Ping LiKey Laboratory of Modern Chinese Medicines and Department of Pharmacognosy, China Pharmaceutical University, Nanjing 210009, People’s Republic of China.E-mail address: lipingli@Tel.: +86-25-8324-2299; 8539-1244; 135********Fax: +86-25-8532-2747AbstractA new HPLC coupled with evaporative light scattering detection (ELSD) method has been developed for the simultaneous quantitative determination of seven major saponins, namely macranthoidinB (1), macranthoidin A (2), dipsacoside B (3), hederagenin-28-O-β-D-glucopyranosyl(6→1)-O-β-D- glucopyranosyl ester (4), macranthoside B (5), macranthoside A (6), and hederagenin-3-O-α-L-arabinopyranosyl(2→1)-O-α-L-rhamnopyranoside (7)in Flos Lonicerae, a commonly used traditional Chinese medicine (TCM) herb.Simultaneous separation of these seven saponins was achieved on a C18 analytical column with a mixed mobile phase consisting of acetonitrile(A)-water(B)(29:71 v/v) acidified with 0.5% acetic acid. The elution was operated from keeping 29%A for 10min, then gradually to 54%B from 10 to 25 min on linear gradient, and then keep isocratic elution with 54%B from 25 to 30min.The drift tube temperature of ELSD was set at 106℃, and with the nitrogen flow-rate of 2.6 l/min. All calibration curves showed good linear regression (r2 0.9922) within test ranges. This method showed good reproducibility for the quantification of these seven saponins in Flos Lonicerae with intra- and inter-day variations of less than 3.0% and 6.0% respectively. The validated method was successfully applied to quantify seven saponins in five sources of Flos Lonicerae, which provides a new basis of overall assessment on quality of Flos Lonicerae.Keywords: HPLC-ELSD; Flos Lonicerae; Saponins; Quantification1. IntroductionFlos Lonicerae (Jinyinhua in Chinese), the dried buds of several species of the genus Lonicera (Caprifoliaceae), is a commonly used traditional Chinese medicine (TCM) herb. It has been used for centuries in TCM practice for the treatment of sores, carbuncles, furuncles, swelling and affections caused by exopathogenic wind-heat or epidemic febrile diseases at the early stage [1]. Though four species of Lonicera are documented as the sources of Flos Lonicerae in China Pharmacopeia (2000 edition), i.e. L. japonica, L. hypoglauca,L. daystyla and L. confusa, other species such as L. similes and L. macranthoides have also been used on the same purpose in some local areas in China [2]. So it is an important issue to comprehensively evaluate the different sources of Flos Lonicerae, so as to ensure the clinical efficacy of this Chinese herbal drug.Chemical and pharmacological investigations on Flos Lonicerae resulted in discovering several kinds of bioactive components, i.e. chlorogenic acid and its analogues, flavonoids, iridoid glucosides and triterpenoid saponins [3]. Previously, chlorogenic acid has been used as the chemical marker for the quality evaluation of Flos Lonicerae,owing to its antipyretic and antibiotic property as well as its high content in the herb. But this compound is not a characteristic component of Flos Lonicerae, as it has also been used as the chemical marker for other Chinese herbal drugs such as Flos Chrysanthemi and so on[4-5]. Moreover, chlorogenic acid alone could not be responsible for the overall pharmacological activities of Flos Lonicerae[6].On the other hand, many studies revealed that triterpenoidal saponins of Flos Lonicerae possess protection effects on hepatic injury caused by Acetaminophen, Cd, and CCl4, and conspicuous depressant effects on swelling of ear croton oil [7-11]. Therefore, saponins should also be considered as one of the markers for quality control of Flos Lonicerae. Consequently, determinations of all types of components such as chlorogenic acid, flavonoids, iridoid glucosides and triterpenoidal saponins in Flos Lonicerae could be a better strategy for the comprehensive quality evaluation of Flos Lonicerae.Recently an HPLC-ELSD method has been established in our laboratory for qualitative and quantitative determination of iridoid glucosides in Flos Lonicerae [12]. But no method was reported for the determination of triterpenoidal saponins in Flos Lonicera. As a series studies on the comprehensive evaluation of Flos Lonicera, we report here, for the first time, the development of an HPLC-ELSD method for simultaneous determination of seven triterpenoidal saponins in the Chinese herbal drug Flos Lonicerae, i.e.macranthoidin B (1), macranthoidin A (2), dipsacoside B (3), hederagenin-28-O-β-D-glucopyranosyl(6→1)-O-β-D- glucopyranosyl ester (4), macranthoside B (5), macranthoside A (6), and hederagenin-3-O-α-L-arabinopyranosyl(2→1)-O-α-L-rhamnopyranoside (7) (Fig. 1).2. Experimental2.1. Samples, chemicals and reagentsFive samples of Lonicera species,L. japonica from Mi county, HeNan province (LJ1999-07), L. hypoglauca from Jiujang county, JiangXi province (LH2001-06), L. similes from Fei county, ShanDong province (LS2001-07), L. confuse from Xupu county, HuNan province (LC2001-07), and L. macranthoides from Longhu county, HuNan province (LM2000-06) respectively, were collected in China. All samples were authenticated by Dr. Ping Li, professor of department of Pharmacognosy, China Pharmaceutical University, Nanjing, China. The voucher specimens were deposited in the department of Pharmacognosy, China Pharmaceutical University, Nanjing, China. Seven saponin reference compounds: macranthoidin B (1), macranthoidin A (2), dipsacoside B (3), hederagenin-28-O-β-D-glucopyranosyl(6→1)-O-β-D- glucopyranosyl ester (4), macranthoside B (5), macranthoside A (6), and hederagenin-3-O-α-L-arabinopyranosyl(2→1)-O-α-L-rhamnopyranoside (7) were isolated previously from the dried buds of L. confusa by repeated silica gel, sephadex LH-20 and Rp-18 silica gel column chromatography, their structures were elucidated by comparison of their spectral data (UV, IR, MS, 1H- NMR and 13C-NMR) with references [13-15]. The purity of these saponins were determined to be more than 98% by normalization of the peak areas detected by HPLC with ELSD, and showed very stable in methanol solution.HPLC-grade acetonitrile from Merck (Darmstadt, Germany), the deionized water from Robust (Guangzhou, China), were purchased. The other solvents, purchased from Nanjing Chemical Factory (Nanjing, China) were of analytical grade.2.2. Apparatus and chromatographic conditionsAglient1100 series HPLC apparatus was used. Chromatography was carried out on an Aglient Zorbax SB-C18 column（250 4.6mm, 5.0µm）at a column temperature of 25℃.A Rheodyne 7125i sampling valve (Cotati, USA) equipped with a sample loop of 20µl was used for sample injection. The analog signal from Alltech ELSD 2000 (Alltech, Deerfield, IL, USA)was transmitted to a HP Chemstation for processing through an Agilent 35900E (Agilent Technologies, USA).The optimum resolution was obtained by using a linear gradient elution. The mobile phase was composed of acetonitrile(A) and water(B) which acidified with 0.5% acetic acid. The elution was operated from keeping 29%A for 10min, then gradually to 54%B from 10 to 25 min in linear gradient, and back to the isocratic elution of 54%B from 25 to 30 min.The drift tube temperature for ELSD was set at 106℃and the nitrogen flow-rate was of 2.6 l/min. The chromatographic peaks were identified by comparing their retention time with that of each reference compound tried under the same chromatographic conditions with a series of mobile phases. In addition, spiking samples with the reference compounds further confirmed the identities of the peaks.2.3. Calibration curvesMethanol stock solutions containing seven analytes were prepared and diluted to appropriate concentration for the construction of calibration curves. Six concentrationof the seven analytes’ solution were injected in triplicate, and then the calibration curves were constructed by plotting the peak areas versus the concentration of each analyte. The results were demonstrated in Table1.2.4. Limits of detection and quantificationMethanol stock solution containing seven reference compounds were diluted to a series of appropriate concentrations with methanol, and an aliquot of the diluted solutions were injected into HPLC for analysis.The limits of detection (LOD) and quantification (LOQ) under the present chromatographic conditions were determined at a signal-to-noise ratio (S/N) of 3 and 10, respectively. LOD and LOQ for each compound were shown in Table1.2.5. Precision and accuracyIntra- and inter-day variations were chosen to determine the precision of the developed assay. Approximate 2.0g of the pulverized samples of L. macranthoides were weighted, extracted and analyzed as described in 2.6 Sample preparation section. For intra-day variability test, the samples were analyzed in triplicate for three times within one day, while for inter-day variability test, the samples were examined in triplicate for consecutive three days. Variations were expressed by the relative standard deviations. The results were given in Table 2.Recovery test was used to evaluate the accuracy of this method. Accurate amounts of seven saponins were added to approximate 1.0g of L. macranthoides,and then extracted and analyzed as described in 2.6 Sample preparation section. The average recoveries were counted by the formula: recovery (%) = (amount found –original amount)/ amount spiked ×100%, and RSD (%) = (SD/mean) ×100%. The results were given in Table 3.2.6. Sample preparationSamples of Flos Lonicerae were dried at 50℃until constant weight. Approximate 2.0g of the pulverized samples, accurately weighed, was extracted with 60% ethanol in a flask for 4h. The ethanol was evaporated to dryness with a rotary evaporator. Residue was dissolved in water, followed by defatting with 60ml of petroleum ether for 2 times, and then the water solution was evaporated, residue was dissolved with methanol into a 25ml flask. One ml of the methanol solution was drawn and transferred to a 5ml flask, diluted to the mark with methanol. The resultant solution was at last filtrated through a 0.45µm syringe filter (Type Millex-HA, Millipore, USA) and 20µl of the filtrate was injected to HPLC system. The contents of the analytes were determined from the corresponding calibration curves.3. Results and discussionsThe temperature of drift tube and the gas flow-rate are two most important adjustable parameters for ELSD, they play a prominent role to an analyte response. In ourprevious work [12], the temperature of drift tube was optimized at 90°C for the determination of iridoids. As the polarity of saponins are higher than that of iridoids, more water was used in the mobile phase for the separation of saponins, therefore the temperature for saponins determination was optimized systematically from 95°C to 110°C, the flow-rate from 2.2 to 3.0 l/min. Dipsacoside B was selected as the testing saponin for optimizing ELSD conditions, as it was contained in all samples. Eventually, the drift tube temperature of 106℃and a gas flow of 2.6 l/min were optimized to detect the analytes. And these two exact experimental parameters should be strictly controlled in the analytical procedure [16].All calibration curves showed good linear regression (r2 0.9922) within test ranges. Validation studies of this method proved that this assay has good reproducibility. As shown in Table 2, the overall intra- and inter-day variations are less than 6% for all seven analytes. As demonstrated in Table 3, the developed analytical method has good accuracy with the overall recovery of high than 96% for the analytes concerned. The limit of detection (S/N=3) and the limit of quantification (S/N=10) are less than 0.26μg and 0.88μg respectively (Table1), indicating that this HPLC-ELSD method is precise, accurate and se nsitive enough for the quantitative evaluation of major non- chromaphoric saponins in Flos Lonicerae.It has been reported that there are two major types of saponins in Flos Lonicerae, i.e. saponins with hederagenin as aglycone and saponins with oleanolic acid as the aglycone [17]. But hederagenin type saponins of the herb were reported to have distinct activities of liver protection and anti-inflammatory [7-11]. So we adoptedseven hederagenin type saponins as representative markers to establish a quality control method.The newly established HPLC-ELSD method was applied to analyze seven analytes in five plant sources of Flos Lonicerae, i.e. L. japonica,L. hypoglauca,L. confusa,L. similes and L. macranthoides(Table 4). It was found that there were remarkable differences of seven saponins contents between different plant sources of Flos Lonicerae. All seven saponins analyzed could be detected in L. confusa and L. hypoglauca, while only dipsacoside B was detected in L. japonica. Among all seven saponins interested, only dipsacoside B was found in all five plant species of Flos Lonicerae analyzed, and this compound was determined as the major saponin with content of 53.7 mg/g in L. hypoglauca. On the other hand, macranthoidin B was found to be the major saponin with the content higher than 41.0mg/g in L. macranthoides,L. confusa, and L. similis, while the contents of other analytes were much lower.In our previous study [12], overall HPLC profiles of iridoid glucosides was used to qualitatively and quantitatively distinguish different origins of Flos Lonicerae. As shown in Fig.2, the chromatogram profiles of L. confusa, L. japonica and L. similes seem to be similar, resulting in the difficulty of clarifying the origins of Flos Lonicerae solely by HPLC profiles of saponins, in addition to the clear difference of the HPLC profiles of saponins from L. macranthoides and L. hypoglauca.Therefore, in addition to the conventional morphological and histological identification methods, the contents and the HPLC profiles of saponins and iridoids could also be used as accessory chemical evidence toclarify the botanical origin and comprehensive quality evaluation of Flos Lonicerae.4. ConclusionsThis is the first report on validation of an analytical method for qualification and quantification of saponins in Flos Lonicerae. This newly established HPLC-ELSD method can be used to simultaneously quantify seven saponins, i.e. macranthoidin B, macranthoidin A, dipsacoside B, hederagenin-28-O-β-D-glucopyranosyl(6→1)-O-β-D- glucopyranosyl ester, macranthoside B, macranthoside A, and hederagenin-3-O-α-L-arabinopyranosyl(2→1)-O-α-L-rhamnopyranoside in Flos Lonicerae. Together with the HPLC profiles of iridoids, the HPLC-ELSD profiles of saponins could also be used as an accessory chemical evidence to clarify the botanical origin and comprehensive quality evaluation of Flos Lonicerae.AcknowledgementsThis project is financially supported by Fund for Distinguished Chinese Young Scholars of the National Science Foundation of China (30325046) and the National High Tech Program（2003AA2Z2010）.[1]Ministry of Public Health of the People’s Republic of China, Pharmacopoeia ofthe People’s Republic of China, V ol.1, 2000, p. 177.[2]W. Shi, R.B. Shi, Y.R. Lu, Chin. Pharm. J., 34(1999) 724.[3]J.B. Xing, P. Li, D.L. Wen, Chin. Med. Mater., 26(2001) 457.[4]Y.Q. Zhang, L.C. Xu, L.P. Wang, J. Chin. Med. Mater., 21(1996) 204.[5] D. Zhang, Z.W. Li, Y. Jiang, J. Pharm. Anal., 16(1996) 83.[6]T.Z. Wang, Y.M. Li, Huaxiyaoxue Zazhi, 15(2000) 292.[7]J.ZH. Shi, G.T. Liu. Acta Pharm. Sin., 30(1995) 311.[8]Y. P. Liu, J. Liu, X.SH. Jia, et al. Acta Pharmacol. Sin., 13 (1992) 209.[9]Y. P. Liu, J. Liu, X.SH. Jia, et al. Acta Pharmacol. Sin., 13 (1992) 213.[10]J.ZH. Shi, L. Wan, X.F. Chen.ZhongYao YaoLi Yu LinChuang, 6 (1990) 33.[11]J. Liu, L. Xia, X.F. Chen. Acta Pharmacol. Sin., 9 (1988) 395[12]H.J. Li, P. Li, W.C. Ye, J. Chromatogr. A 1008(2003) 167-72.[13]Q. Mao, D. Cao, X.SH. Jia. Acta Pharm. Sin., 28(1993) 273.[14]H. Kizu, S. Hirabayashi, M. Suzuki, et al. Chem. Pharm. Bull., 33(1985) 3473.[15]S. Saito, S. Sumita, N. Tamura, et al. Chem Pharm Bull., 38(1990) 411.[16]Alltech ELSD 2000 Operating Manual, Alltech, 2001, p. 16. In Chinese.[17]J.B. Xing, P. Li, Chin. Med. Mater., 22(1999) 366.Fig. 1 Chemical structures of seven saponins from Lonicera confusa macranthoidin B (1), macranthoidin A (2), dipsacoside B (3), hederagenin-28-O-β-D-glucopyranosyl(6→1)-O-β-D- glucopyranosyl ester (4), macranthoside B (5), macranthoside A (6), and hederagenin-3-O-α-L-arabinopyranosyl(2→1)-O-α-L-rhamnopyranoside (7)Fig. 2Representative HPLC chromatograms of mixed standards and methanol extracts of Flos Lonicerae.Column: Agilent Zorbax SB-C18 column（250 4.6mm, 5.0µm）, temperature of 25℃; Detector: ELSD, drift tube temperature 106℃, nitrogen flow-rate 2.6 l/min.A: Mixed standards, B: L. confusa, C: L. japonica, D: L. macranthoides, E: L. hypoglauca, F: L. similes.Table 1 Calibration curves for seven saponinsAnalytes Calibration curve ar2Test range(μg)LOD(μg)LOQ(μg)1 y=6711.9x－377.6 0.9940 0.56–22.01 0.26 0.882 y=7812.6x－411.9 0.9922 0.54–21.63 0.26 0.843 y=6798.5x－299.0 0.9958 0.46–18.42 0.22 0.724 y=12805x－487.9 0.9961 0.38–15.66 0.10 0.345 y=4143.8x－88.62 0.9989 0.42–16.82 0.18 0.246 y=3946.8x－94.4 0.9977 0.40–16.02 0.16 0.207 y=4287.8x－95.2 0.9982 0.42–16.46 0.12 0.22a y: Peak area; x: concentration (mg/ml)Table 2 Reproducibility of the assayAnalyteIntra-day variability Inter-day variability Content (mg/g) Mean RSD (%) Content (mg/g) Mean RSD (%)1 46.1646.2846.2246.22 0.1346.2245.3647.4226.33 2.232 5.385.385.165.31 2.405.285.345.045.22 3.043 4.374.304.184.28 2.244.284.464.024.255.204 nd1)-- -- nd -- --5 1.761.801.821.79 1.701.801.681.841.77 4.706 1.281.241.221.252.451.241.341.201.26 5.727 tr2)-- -- tr -- -- 1): not detected; 2): trace. RSD (%) = (SD/Mean) ×100%Table 3 Recovery of the seven analytesAnalyteOriginal(mg) Spiked(mg)Found(mg)Recovery(%)Mean(%)RSD(%)1 23.0823.1423.1119.7122.8628.1042.7346.1351.0199.7100.699.399.8 0.722.692.672.582.082.913.164.735.515.7698.197.6100.698.8 1.632.172.152.091.732.182.623.884.404.6598.8103.297.799.9 2.94nd1)1.011.050.980.981.101.0297.0104.8104.1102.0 4.250.880.900.910.700.871.081.561.752.0197.197.7101.898.9 2.660.640.620.610.450.610.751.081.211.3397.796.796.096.8 0.97tr2)1.021.101.081.031.111.07100.9102.799.1100.9 1.81): not detected; 2): trace.a Recovery (%) = (Amount found –Original amount)/ Amount spiked ×100%, RSD (%) = (SD/Mean) ×100%Table 4 Contents of seven saponins in Lonicera spp.Content (mg/g)1 2 3 4 5 6 7 L. confusa45.65±0.32 5.13±0.08 4.45±0.11tr1) 2.04±0.04tr 1.81±0.03 L. japonica nd2)nd 3.44±0.09nd nd nd nd L. macranthoides46.22±0.06 5.31±0.13 4.28±0.10 tr 1.79±0.03 1.25±0.03 tr L. hypoglauca11.17±0.07 nq3)53.78±1.18nd 1.72±0.02 2.23±0.06 2.52±0.04 L. similes41.22±0.25 4.57±0.07 3.79±0.09nd 1.75±0.02tr nd 1): trace; 2): not detected.. 3) not quantified owing to the suspicious purity of the peak.。
恩格列净联合西格列汀治疗老年2_型糖尿病患者的临床疗效分析
·药物与临床·糖尿病新世界 2023年3月DOI：10.16658/ki.1672-4062.2023.05.059恩格列净联合西格列汀治疗老年2型糖尿病患者的临床疗效分析臧道军，龚红燕江苏省常州市德安医院老年内科，江苏常州213000[摘要]目的探讨老年2型糖尿病患者使用恩格列净+西格列汀治疗的临床效果。
方法选取2020年1月—2021年12月常州市德安医院接诊的100例老年2型糖尿病患者作为研究对象，根据不同用药方式分为对照组与研究组，各50例，对照组接受西格列汀治疗，研究组接受恩格列净+西格列汀治疗，就两组患者血糖指标、炎性指标、胱抑素C（Cys-C）、血尿素氮（BUN）、血同型半胱氨酸（Hcy）指标进行比较。
结果治疗前两组血糖指标相比，差异无统计学意义（P>0.05），治疗后，研究组HbA1c、FPG及2 hPG明显低于对照组，差异有统计学意义（P<0.05）；治疗前两组炎性指标比较，差异无统计学意义（P>0.05），治疗后，研究组IL-4、IL-6及TNF-α明显低于对照组，差异有统计学意义（P<0.05）；治疗前两组Cys-C、BUN及Hcy相比，差异无统计学意义（P>0.05），治疗后，研究组患者Cys-C、BUN及Hcy明显低于对照组，差异有统计学意义（P<0.05）。
结论对于老年2型糖尿病患者开展恩格列净+西格列汀治疗能有效改善血糖指标，降低Hcy，提升肾功能，治疗效果显著。
[关键词] 老年人群；恩格列净；西格列汀；2型糖尿病[中图分类号] R4 [文献标识码] A [文章编号] 1672-4062（2023）03（a）-0059-04Clinical Efficacy Analysis of Empagliflozin Combined with Sitagliptin in the Treatment of Elderly Patients with Type 2 Diabetes MellitusZANG Daojun, GONG HongyanDepartment of Geriatric Medicine, Changzhou De'an Hospital, Changzhou, Jiangsu Province, 213000 China[Abstract] Objective To investigate the clinical effect of treatment with empagliflozin + sitagliptin in elderly patients with type 2 diabetes mellitus.Methods A total of 100 elderly patients with type 2 diabetes mellitus admitted to Chang⁃zhou De'an Hospital from January 2020 to December 2021 were selected as study subjects. The cases were divided into control group and study group according to different medication administration, fifty cases in each. The control group received sitagliptin treatment and the study group received empagliflozin + sitagliptin treatment. The blood glu⁃cose index, inflammatory index, cystatin C (Cys-C), blood urea nitrogen (BUN), and blood homocysteine (Hcy) index were compared between the two groups.Results There was no statistically significant difference in blood glucose in⁃dexes between the two groups before treatment (P>0.05). After treatment, HbA1c, FPG and 2 hPG of the study group were significantly lower than those in the control group, the difference was statistically significant (P<0.05). There was no statistically significant difference in inflammatory indexes between the two groups before treatment (P>0.05). After treatment, IL-4, IL-6 and TNF-α in the study group were significantly lower than those in the control group, the dif⁃ference was statistically significant (P<0.05). There was no statistically significant difference in the Cys-C, BUN and Hcy between the two groups before treatment (P>0.05). After treatment, the Cys-C, BUN and Hcy of the study group were significantly lower than those in the control group, the difference was statistically significant (P<0.05).Conclusion For elderly patients with type 2 diabetes mellitus, treatment with empagliflozin + sitagliptin can effec⁃tively improve blood glucose index, reduce Hcy and enhance renal function, with significant therapeutic effects.[作者简介]臧道军（1974-），男，本科，副主任医师，研究方向为老年内科。
USP微生物限度检查 中文
USP微生物限度检查中文61）微生物限度检测（MICROBIAL LIMIT TESTS）此章提供方法来检测可能存在的好氧微生物其他制药过程中可能出现的微生物的数量，包括原材料和成品中的。
如果经过验证确认可以得到相同或更好的检测结论，也允许采用自动化的检测方法。
在样品检测过程中须进行无菌操作。
若无特别说明，则“培养（incubate）”一词指在30—35℃的培养箱内培养24至48小时；“生长（growth）”一词用于专门的判定，说明“存在和可能存在活的微生物”。
准备实验 (Preparatory Testing)本章涉及实验结果的有效性取决于：提供的被检测样品本身在实验条件下，被充分证明不会抑制可能存在的微生物的生长。
因此，在准备样品时，需要正规的实验操作和符合要求的实验条件，接种稀释样品到含有以下（微生物）培养物的培养基：金黄色（奥里斯）葡萄球菌（Staphylococcus aureus）,大肠埃希氏菌（Escherichia coli）, 铜绿假单胞菌(Pseudomonas aeruginosa), 和沙门氏菌（Salmonella）。
方法如下：将用肉汤培养基培养24小时后的（微生物）不小于10-3稀释的微生物培养物，加1 ml（微生物）培养液到磷酸(盐)缓冲液(pH 7.2)，液体大豆酪蛋白消化物培养基（Fluid Soybean-Casein Digest Medium），或者液体乳糖培养基（Fluid Lactose Medium）。
相应培养基培养失败则需要采取以下方法更改检测程序：（1）增加稀释液体积，检测样品加入量仍维持不变；或者（2）中和一定数量的干扰因子；或者（3）结合（1）、（2）得出适当条件，使接种物得以生长。
以下是一些物质的成分和浓度，该物质及浓度可用于加入培养基、阻止物质发挥抑菌作用：大豆卵磷脂（soy lecithin, 0.5%）或者聚山梨醇酯20（polysorbate 20, 4.0%）。
重组贻贝粘蛋白的表征及功效评价
生物技术进展 2023 年 第 13 卷 第 4 期 596 ~ 603Current Biotechnology ISSN 2095‑2341研究论文Articles重组贻贝粘蛋白的表征及功效评价李敏 ， 魏文培 ， 乔莎 ， 郝东 ， 周浩 ， 赵硕文 ， 张立峰 ， 侯增淼 *西安德诺海思医疗科技有限公司，西安 710000摘要：为了推进重组贻贝粘蛋白在医疗、化妆品领域的应用，对大肠杆菌规模化发酵及纯化生产获得的重组贻贝粘蛋白进行了表征及功效评价。
经Edman 降解法、基质辅助激光解吸电离飞行时间质谱、PITC 法、非还原型SDS -聚丙烯酰胺凝胶电泳法、凝胶法、改良的Arnow 法对重组贻贝粘蛋白进行氨基酸N 端测序、相对分子量分析、氨基酸组成分析、蛋白纯度分析、内毒素含量测定、多巴含量测定；通过细胞迁移、斑马鱼尾鳍修复效果对重组贻贝粘蛋白进行功效评价。
结果显示，获得的重组贻贝粘蛋白与理论的一级结构一致，蛋白纯度达95%以上，内毒素<10 EU ·mg -1,多巴含量大于5%；重组贻贝粘蛋白浓度为60 μg ·mL -1时能够显著促进细胞增殖的活性（P <0.01）；斑马鱼尾鳍面积样品组与模型对照组相比极显著增加（P <0.001）。
研究结果表明，重组贻贝粘蛋白具有显著的促细胞迁移和修复愈合的功效，具备作为生物医学材料的潜质。
关键词：贻贝粘蛋白；基因重组；生物材料；表征；功效评价DOI ：10.19586/j.20952341.2023.0021 中图分类号：S985.3+1 文献标志码：ACharacterization and Efficacy Evaluation of Recombinant Mussel Adhesive ProteinLI Min ， WEI Wenpei ， QIAO Sha ， HAO Dong ， ZHOU Hao ， ZHAO Shuowen ， ZHANG Lifeng ，HOU Zengmiao *Xi'an DeNovo Hith Medical Technology Co.， Ltd ， Xi'an 710000， ChinaAbstract ：In order to promote the application of recombinant mussel adhesive protein in the medical and cosmetics field ， the recombi⁃nant mussel adhesive protein obtained from scale fermentation and purification of Escherichia coli was characterized and its efficacy was evaluated. Amino acid N -terminal sequencing ， relative molecular weight analysis ， amino acid composition analysis ， protein purityanalysis ， endotoxin content ， dihydroxyphenylalanine （DOPA ） content of recombinant mussel adhesive protein were determined by the following methods ： Edman degradation ， matrix -assisted laser desorption ionization time -of -flight mass spectrometry （MALDI -TOF -MS ）， phenyl -isothiocyanate （PITC ）， nonreductive SDS -polyacrylamide gel electrophoresis （SDS -PAGE ）， gel method ， modified Ar⁃now. The efficacy of recombinant mussel adhesive protein was evaluated by cell migration and repairing effect of zebrafish tail fin. Re⁃sults showed that the obtained recombinant mussel adhesive protein was confirmed to be consistent with the theoretical primary structure ， protein purity of more than 95%， endotoxin <10 EU ·mg -1， DOPA content above 5%. When the recombinant mussel adhesive protein concentration was 60 μg ·mL -1， the effect of promoting cell proliferation was the most obvious ， and it had very significant activity （P <0.01）. The caudal fin area of zebrafish in sample group was significantly increased compared with model control group （P <0.001）. The results indicated that recombinant mussel adhesive protein can promote cell migration and repair healing and has the potential to be used as biomedical materials.Key words ：mussel adhesive protein ； gene recombination ； biological materials ； representation ； efficacy evaluation贻贝粘蛋白（mussel adhesive protein ， MAP ）也称作贻贝足丝蛋白（mussel foot protein ，Mfps ），收稿日期：2023⁃02⁃24； 接受日期：2023⁃03⁃31联系方式：李敏 E -mail:*******************;*通信作者 侯增淼 E -mail:***********************.cn李敏，等：重组贻贝粘蛋白的表征及功效评价是海洋贝类——紫贻贝（Mytilus galloprovincalis）、厚壳贻贝（Mytilus coruscus）、翡翠贻贝（Perna viri⁃dis）等分泌的一种特殊的蛋白质，贻贝中含有多种贻贝粘蛋白，包括贻贝粘蛋白（Mfp 1～6）、前胶原蛋白（precollagens）和基质蛋白（matrix proteins）等［1］。
迷迭香酸对过氧化氢处理下的皮肤黑色素瘤的抗氧化作用(原文翻译)
迷迭香酸（罗丹酚酸）对H2O2处理过的皮肤黑色素瘤细胞的抗氧化作用Sun Mi Yoo1 and Jeong Ran Kang2*1.韩国光州500-741号东冈大学美容系2.韩国首尔143-701号建国大学生物工程系2009.2.6收到 2009.4.17接收本学科旨在检测迷迭香酸对人工孵育的皮肤黑色素瘤细胞在ROS下的抗氧化作用。
通过XTT比色法，以细胞毒性和抗氧化作用来分析细胞粘附活性，DPPH自由基清除活性以及H2O2处理1-10h和未经处理的两种情况下乳酸脱氢酶的活性。
用20-110 μM 的H2O2处理皮肤黑色素瘤细胞5-7h后，细胞活性的降低呈剂量和时间依赖性。
通过XTT比色法测得H2O2的半抑制浓度（IC50 ）为90μM。
同时H2O2增强了LDH细胞的剂量依赖性。
用50-90μM的H2O2处理8h后测得LDH50为60 μM H2O2。
迷迭香酸能增强细胞活性和DPPH自由基清除活性，降低乳酸盐脱氢酶的活性。
细胞的H2O2处理证实了对人工孵育的皮肤黑色素瘤细胞的强抗氧化作用。
通过H2O2的处理，迷迭香酸能在细胞内能增强细胞活性和DPPH 自由基清除活性，降低乳酸盐脱氢酶的活性。
这被认为是迷迭香酸对ROS（ROS）如H2O2的抗氧化作用。
Key words：DPPH-radical scavenging, LDH, rosmarinic acid, XTT assay关键字：DPPH自由基清除活性，乳酸脱氢酶，迷迭香酸，XTT比色法据研究发现，ROS通过氧化应激对细胞的损伤和一些脑部疾病比如帕金森症或心脏疾病例如心肌梗塞之间有很大的关联[Difazio et al., 1992; Delanty and Dichter, 1998].尤其是研究人员认为ROS是皮肤老化的一个主要的因素后，一直试图从ROS方面研究衰老。
[Yokozawa et al., 1998].据研究表明，ROS的氧化应激会通过萎缩细胞引起各种疾病，例如超氧自由基、H2O2（H2O2）或羟基自由基的巯基蛋白反应中断酶的活性，破坏脱氧RMA（DNA）或RMA（RNA），诱导细胞膜脂质过氧化。
谷胱甘肽促进液体发酵体系中蛹虫草合成虫草素
Abstract: C o r d y c e p i n is the m a i n active ingredient of the medicinal f u n g u s C . m ilitoris, with a variety of physiological functions s u c h as anti-cancer, anti-tumor a n d anti-virus activity.Oxidative stress w a s s h o w n to b e involved in the regulation of s e c o n d a r y m e t a b o l i s m of fila m e n t o u s fungi, h o w e v e r ,the relationship b e t w e e n oxidative stress a n d the regulation of cordycepin m e t a b o l i s m in C . militaris has not yet reported so far. In this study, to investigate the influence of oxidative stress o n c o r d ycepin m e t a b o l i s m , glutathione (G S H ), as the antioxidant for regulating cellular red o x state, w a s s u p p l e m e n t e d during liquid s u b m e r g e d fermentation of C . m ilitaris. T h e experimental data s h o w e d that the yield of cordycepin could reach (439.69±12.43)m g / L in 20 days w h e n 3.0g/L G S H w a s a d d e d to the m e d i u m , 471.24% higher t h a n that of t h e control (without addition of G S H ). T h e activity of glutathione peroxidase (G P X ) increased b y 414.82% as c o m p a r e d with t h e control. T h e relative g e n e expression levels of C n s l a n d Cns2 indicated by q R T - P C R w e r e significantly up-regulated by s u p p l e m e n t a t i o n of G S H , increasing b y 540.67 times a n d 25.81 t i m e s as c o m p a r e d with the control in 15 d a y s , respectively.T h e e x p e r imental results
国际上著名的从事药剂学研究的专家
Intra Oral Delivery (口腔内传递)直接由口腔黏膜吸收,瞬间进入血液循环,有效成分不流失。
Universities, Departments,FacultiesResearchersButler University College of Pharmacy and Health Sciences Health Sciences USA Associate Professor Nandita G. DasMain focus on her research facilities are about peformulation, biopharmaceutics, drug targeting, anticancer drug delivery.Purdue University School of Pharmacy and Pharmacal Sciences Department of Industrial and Physical Pharmacy (IPPH) USA Professor Kinam ParkControlled Drug Delivery, Glucose-Sensitive Hydrogels for Self-Regulated Insulin Delivery, Superporous Hydrogel Composites, Oral Vaccination using Hydrogel Microparticles, Fractal Analysis of Pharmaceutical Solid Materials.St. John's University School of Pharmacy and Allied Health ProfessionsUSA Professor Parshotam L. MadanControlled and targeted drug delivery systems; Bio-erodible polymers as drug delivery systemsThe University of Iowa College of Dentistry Department of Oral Pathology, Radiology, and Medicine USA Professor Christopher A. Squierpermeability of skin, and oral mucosa to exogenous substances, including alcohol and tobacco, and drug deliveryThe University of Iowa College of Pharmacy Department of Pharmaceutics USA Associate Professor Maureen D. DonovanMucosal drug delivery especially via the nasal, gastrointestinal and vaginal epithelia; and mechanisms of drug absorption and disposition.The University of Texas at San Antonio College of Engineering Department of Biomedical Engineering USA Professor Jeffrey Y. ThompsonDental restorative materials and implantsThe University of Utah Pharmaceutics & Pharmaceutical Chemistry USA Professor John W. MaugerDr. Maugner is mainly focused on dissolution testing and coating technology of orally administered drug products with bitter taste about which he is one of the inventors of a filed patent.University of Kentucky College of Pharmacy Pharmaceutical Sciences USA Professor Peter CrooksDr. Crooks is internationally known for his research work in drug discovery, delivery, and development, which includes drug design and synthesis, pharmacophore development, drug biotransformation studies, prodrug design, and medicinal plant natural product research. His research also focuses on preclinical drug development, including drug metabolism and pharmacokinetics in animal models, dosage form development, and drug delivery assessment using both conventional and non-conventional routes, and preformulation/formulation studies.Associate Professor Russell MumperDr. Mumper's main research areas are thin-films and mucoadhesive gels for (trans)mucosal delivery of drugs, microbicides, and mucosal vaccines, and nanotemplate engineering of nano-based detection devices and cell-specific nanoparticles for tumor and brain targeting, gene therapy and vaccines.West Virginia University School of Pharmacy Department of Basic Pharmaceutical Sciences USA Associate Professor Paula Jo Meyer StoutDr. Stout's research areas are composed of dispersed pharmaceutical systems, sterile product formulation DDS for dental diseases and coating of sustained release formulations.Monash University Victorian College of Pharmacy Department of Pharmaceutics Australia Professor Barrie C. FinninTransdermal Drug Delivery. Physicochemical Characterisation of Drug Candidates. Topical Drug Delivery. Drug uptake by the buccal mucosaProfessor Barry L. ReedTransdermal Drug Delivery. Topical Drug Delivery. Formulation of Dental Pharmaceuticals.University of Gent Faculty of Pharmaceutical Sciences Department of Pharmaceutics Belgium Professor Chris Vervaet-Extrusion/spheronisation - Bioadhesion - Controlled release based on hot stage extrusion technology - Freeze-drying - Tabletting and - GranulationPh.D. Els AdriaensMucosal drug delivery (Vaginal and ocular) Nasal BioadhesionUniversity of Gent Faculty of Pharmaceutical SciencesLaboratory of Pharmaceutical Technology Belgium Professor Jean Paul Remonbioadhesive carriers, mucosal delivery, Ocular bioerodible minitablets, Compaction of enteric-coated pellets; matrix-in-cylinder system for sustained drug delivery; formulation of solid dosage forms; In-line monitoring of a pharmaceutical blending process using FT-Raman spectroscopy; hot-melt extruded mini-matricesDanish University of Pharmaceutical Sciences Department of Pharmaceutics Denmark Associate Professor Jette JacobsenLow soluble drugs ?in vitro lymphatic absorption Drug delivery to the oral cavity ?in vitro models (cell culture, diffusion chamber) for permeatbility and toxicity of drugs, in vivo human perfusion model, different formulation approaces, e.g. iontophoresis.。
氰基硼氢化钠还原胺化京尼平合成拟生物碱与活性
氰基硼氢化钠还原胺化京尼平合成拟生物碱与活性秦杰琛 1），曾小娟 1），张韶湘 1），张晓梅 2），刘鑫洋 1），邹 澄 1），赵 庆 2）（1）昆明医科大学药学院暨云南省天然药物药理重点实验室，云南 昆明 650500；2）云南中医药大学中药学院，云南 昆明 650500）[ 摘要 ] 目的 以京尼平苷为原料通过还原胺化反应合成拟生物碱的方法。
方法 京尼平与胺类化合物在氰基硼氢化钠存在下进行还原反应：京尼平与芳基乙胺的甲醇溶液混合后，加入过量氰基硼氢化钠，放置室温下反应3d，产物经石油醚-异丙醇-二乙胺，石油醚-乙酸乙酯等洗脱分离。
结果 合成共得到9个拟生物碱并对部分拟生物碱进行活性筛选，找到治疗Ⅱ型糖尿病的PTP1B 抑制剂。
结论 部分受试化合物对PTP1B 有抑制作用。
一系列活性衍生物的获得为化合物结构及其生物活性间的构效关系研究打下了基础，有利于寻找具有更高活性的PTP1B 抑制剂。
[ 关键词 ] 京尼平； 拟生物碱； 还原胺化； 抗PTP1B 活性[ 中图分类号 ] R284.1 [ 文献标志码 ] A [ 文章编号 ] 2095 − 610X （2021）02 − 0018 − 05Reductive Amination of Genipin with NaBH 3CN toSynthesize Alkaloid-likes and BioactivityQIN Jie-chen 1），ZENG Xiao-juan 1），ZHANG Shao-xiang 1），ZHANG Xiao-mei 2），LIU Xin-yang 1），ZOU Cheng 1），ZHAO Qing 2）（1） School of Pharmaceutical Science & Yunnan Key Laboratory of Pharmacology for Natural Products ，Kunming Medical University ，Kunming Yunnan 650500; 2） Faculty of Pharmacy ，Yunnan University of Chinese Medicine ，Kunming Yunnan 650500，China ）［Abstract ］ Objective To explore a method for the synthesis of alkaloid-likes from Genipin by reductive amination is reported. Methods The reduction of Genipin and amines in the presence of sodium cyanoborohydride：after the methanol solution of Genipin and arylethylamine was mixed，excessive sodium cyanoborohydride was added and the reaction was kept at room temperature for 3 days. The product was eluted and separated on silica gel by petroleum ether-isopropyl alcohol-diethylamine and petroleum ether-ethyl acetate. Results Nine alkaloid-likes were synthesized. Some alkaloid-likes were screened for inhibition activity of PTP1B enzyme for Ⅱ diabetes treatment. Conclusions All of the tested compounds have a certain inhibitory effect on PTP1B. The acquisition of a series of active derivatives has laid a foundation for the study of the structure-activity relationship between the compounds and their bioactivities，so as to facilitate the search for more active PTP1B inhibitors.［Key words ］ Genipin；Alkaloid-likes；Reductive amination；Inhibition activity against PTP1B目前，通过发现先导化合物是现代新药研发的重要出发点，研究者对具有特定生物活性的先导化合物，利用生物、化学方法进行结构修饰，从而减少化合物毒副作用，提高化合物的活性，增强其生物利用度，最终找到一些作用效果明显，副作用少的新药应用于临床治疗。
红参中的一种新氨基酸衍生物_英文_
A N e w Amino Acid Derivative from R ed Ginseng3Y i2Nan Zheng1,H Okuda2,Li2Kun Han1,Lan Xiang1,Y Matsuura2,T Takaku2and K Kameda21.Jili n A gricult ural U niversity,Changchun130118;2.School of Medici ne,Ehi me U niversity,Japan791202Received J uly17,1996;Accepted October13,1997 Abstract Five ninhydrin2positive compounds were isolated from the water extract of red ginseng through the Bio2gel P22column chromatography.One compound is known as arginine.Another compound is a new substance which is determined as1′2Nα2arginine21′2deoxy24′2O2(a2D2glucopyranosyl)2D2fructose (AF G)through the analysis of13C NMR,1H NMR,IR and UV s pectra.Its structure is also verified through the reaction of maltose and arginine.The structures of com pounds3,4and5are being re2 searched.K ey w ords Red ginseng;Argininyl2fructosyl2glucose(AF G)IntroductionRed ginseng is a processed product of Panax gi nseng C.A.Meyer with a long histo2 ry of medical application.Much work has been reported about ginseng saponins,polysaccha2 rides and so on,however,little has been done so far about their ninhydrin2positive substance (nonprotein amino acid)(1,2).We isolated five ninhydrin2positive compounds from the outer dialysate of water extract of red ginseng through the Bio2gel P22column chromatogra2 phy,among which compound2was a new substance.In this paper its isolation and iden2 tification are pound2,UVλmax 201nm(terminal absorption).IR(K Br) 167821(C=O).Its peak retention time was 130.92′on amino acid pound2 liberated arginine,glucose and fructose after alkaline hydrolysis.Analysis of13C NMR spec2 trum suggested the presence of18carbons (Table1).There were12carbons in the sac2 charide moieties,the chemical shifts atδ101. 38,δ96.19indicated the presence of two sac2 charide moieties.In1H NMR spectrum,the anomeric proton signal possessed a coupling constant of J<4Hz,and glucose was obtained after enzymatic hydrolysis,thus glucose should be the end saccharide.DEPT spectrum showed that compound2hold three quaternary car2 bons,six methylene and nine methine groups. The structure of compound2has thus been suggested as argininyl2fructosyl2glucose (AF G).It is reported(3,4)that free fructose exists as a mixture of4isomers(α2pyranose,β2pyra2 nose,α2furanose,β2furanose)in water solu2 tion and theα2pyranose form is negligible in comparison with the other three forms.The13 C NMR spectrum of compound2showed the presence of three isomers.In1H NMR spec2 trum1″2H split into three double peaks,δ5.3First published in Chinese in Acta Pharmaceutica Sinica,1996,31(3)191～195.00(d ,J =3.66Hz ),δ5.55(d ,J =3.66Hz )and d 5.09(d ,J =3.66Hz ).The ratio of in 2ten 2sity was 1:1:4,which was probably influ 2enced by the three isomers of the fructose moi 2ety in the forms of α2furanose ,β2furanose andβ2pyranose.All these results demonstrated that the glucose unit could not be linked at the position C 22′C 25′or C 26′of the fructose.The 13C NMR spectrum showed about 10ppm up 2field shift for C 21′of fructose ,suggesting the C 21′hydroxyl group of the fructose was substi 2tuted by the amino group of arginine and thus produced C 2N bond (5),so the C 21″of glucosecould only be connected with C 23′or C 24′offructose.According to the principle of Maillard re 2action(6,7),we reacted arginine with maltose.The resulting product showed identical Rf ,MW and 1H NMR with those of compound 2.Consequently ,compound 2was charac 2terized as 1′2N α2arginine 21′2deoxy 24′2O 2(α2D 2glucopyranosyl )2D 2fructose (AF G ).Its struc 2ture is shown in Fig.1.Fig.1.Chemical structure of argininyl 2fructosylglu 2cose (AFG,the predominant form).The physiological research indicated that AF G significantly inhibited the maltase activity of the intestine and promoted spleen cell prolif 2eration ,it also had immunity effect.The de 2tails of its physiological effect will be reported soon.The content of AF G in red ginseng is 5.37%,while only 1.75%in the white ginseng.ExperimentalUV spectrum ,DU 27500spectropho 2tometer ;IRspectrum ,F TS 27F T 2Infraredspectrometer ;1H NMR and 13C NMR spectra :Jeol Gsx 2270spectrometer (D2O ,25℃270and 67.8MHz respectively );Molecular weight ,laser desorption ionization 2MS (Shimadzu Kratos K ompact MALDE III );GC 2MS ,Shimadzu Q P 21000Mass 2spec 2trome 2ter ;Analysis of amino acid ,Hitachi 2835amino acid analyzer ;Analysis of elements ,Carlo Er 2ba 1106elemental analyzer.Red ginseng was obtained from Ai 2Lin G inseng Farm.Extraction and isolationRed ginseng powder (100g )was soaked in 5volumes of MeOH with stirring for 24h at 25℃,the residue was then extracted with wa 2ter for 3times ,each time using 10times of water at 4℃for 12h.The combined extract was centrifuged for 30min (8000rpm )and the resultant supernatant was concentrated to a suitable volume for dialysis against water.The outer dialysates were concentrated and freeze 2dried.The resulting powder (9g )was dis 2solved in water (80mg/ml ),then subjected to Bio 2gel P 22column (2.7×90cm ),eluted with water (eluting rate :18ml/h ).Eluates were collected in 100tubes with an auto frac 2tion 2collector (3ml per tube ).The ninhydrin 2positive eluates were analyzed with Hitachi 2835amino acid analyzer.The solutions ob 2tained at retention time 131.28′were combined ,con 2centrated and developed on preparative TLCplate to obtain pure compound2(40mg).Identif icationCompound2.White powder,mp158～160℃(dec),purple in ninhydrin reaction, molecular weight:498.Elemental analysis for C18H34N4O12:calc.C43.38,H6.88,N11. 24;found C43.84,H 6.24,N11.08. HPTLC(n2butanol2acetic acid2water,2:1:1) Rf0.20.No significant absorption in UV spectrum.IR(K Br)cm21:3425(OH), 2928,2858(CH),1678(C=O),1633(C= N).DEPT indicated that compound2had3 methyl,6methylene and9methine groups, 1H NMR:1.64(m,2H),1.90(m,2H),3. 16(t,J=6.72Hz,2H),5.00(d,J=3.66 Hz),5.05(d,J=3.66Hz),5.09(J=3.66 Hz).The ratio of intensity was1:1:4.13C NMR is shown in Table1.T able1.13C NMR chemical shifts of compound2in D2OCδppm Cδppm Cδppm1173.50(s)1′53.13(t)1″101.38(d) 263.13(d)2′96.19.(s)2″72.49(d) 327.26(t)3′69.72.(d)3″73.55(d) 424.71(t)4′78.38(d)4″70.36(d) 541.23(t)5′70.02.(d)5″73.19(d) 6157.57(s)6′64.76(t)6″61.30(t) Alkaline hydrolysis:compound2(30mg) was treated with3mol・L21N H4OH(30ml) at100℃for1h.The reaction mixture was isolated and purified on Bio2gel P22column, giving ninhydrin2positive substance(5mg) and carbohydrate(9mg).The ninhydrin2positive substance was ana2 lyzed by TLC and amino acid analyzer,the Rf and retention time were identical with those of authentic arginine.13C NMR:184.84(2COOH),160.11(H2N2C=N H),58.67(2 CH2COOH),44.22(2CH22N H),34.16 (CH22CH2),27.69(CH22CH22CH2).1H NMR:1.50(m,2CH22),1.53(m,2CH22), 3.10(t,J=4.88Hz,2CH22N H),3.17(t,J =4.88Hz,N H22CH2COOH).All these data were the same as those of authentic arginine.The carbohydrate moiety(3mg)of com2 pound2was treated with0.8μl of TMS reα2 gent and stirred for30sec.Then the reaction mixture was let to stand for5min and centri2 fuged at3000rpm.The supernatant was anα2 lyzed with GC2MS.The retention times of those trimethylsilylated carbohydrates were i2 dentical with those of authentic trimethylsily2 lated glucose and fructose.So the carbohy2 drate moiety of compound2was identified as glucose and fructose.Enzymatic hydrolysis:after incubation of 0.2mg of compound2with0.1U ofα2gluco2 sidase in40μl of10mmol・L21phosphate buffer(p H6.8)containing15mmol・L21ED2 TA at37℃for1h,the reaction mixture was analyzed with GC2MS and TLC.The retention times were16.25◊and21.25◊on GC2MS which were identical with those of authentic trimethylsilylated glucose.The reaction mix2 ture was developed on TLC plate(silica gel, Merck Co.),developing system:isopropyl al2 cohol2acetone20.1mol・L21sodium lactate(4: 4:2),Rf0.46.It was identical with that of authentic glucose.Chemical synthesis(8)L2arginine(1.9g)and maltose(4g) were dissolved in80ml of glacial acetic acid, stirred for1h at80℃.The reaction mixture was subjected to Bio2gel P22column,and yielded200mg of white powder.HPTLC n2butanol2acetic acid2water,2:1:1,Rf0.20; mp156～159℃and mixed mp with compound 2was not depressed;molecular weight,498. Amino acid analysis:t R,131.06′.1H NMR: 1.64(m,2H),1.90(m,2H),3.16(t,J= 6.72Hz,2H),5.00(d,J=3.66Hz),5.05(d,J=3.66Hz)and5.09(d,J=3.66Hz). The intensity ratio of the former three values was1:1:4,and that of the latter three was the same.All these data were identical with those of compound2Acknow ledgements Infrared spectrum was analyzed by the Changchun Applied Chemical Insti2 tute.UV spectrum was analyzed by Analysis Center of Jilin Agricultural University.1H NMR and 13C NMR were analyzed by Analysis Center of Ehime University,Japan.R eferences1Li XG,Zheng YN,Wei CY.J Jilin Agricultural University,1989,11:322Y ang L,Y e YH,Yuan HS,Xing Q Y.Chi n Sci B ull,1991,36:5133Doddrell D,Allerhand A.J A m Chem Soc,1971,93:27794Allerhand A.Pure A ppl Chem,1975,41:2475Chen DC.C2N M R and its A pplication i n Chemist ry of Chi nese M aterial Herb and Drug.Beijing:Peoples◊Hygiene Publishing House,1991:2816¨Oste R,Sj¨o din P,J¨аgerstad M.Food Chemist ry,1985,16:377Hayase F.化学と生物,1993,31:5928Einarsson H.The mode of action of antibacterial Maillard reaction products.The M aillard reaction advances i n lif e sciences. Basel:Birkh¨аuser Verlog,1990:215红参中的一种新氨基酸衍生物郑毅男 奥田拓道 韩立坤 向兰松浦幸永 高久武司 龟田健治1.吉林农业大学,长春130118;2.爱媛大学医学部,791202,日本 摘要 红参水提液经聚丙烯酰胺(Bio2gel P22)柱层析,分离得到五个茚三酮反应阳性物质。
幽门螺杆菌感染内镜下胃黏膜表现与14C_尿素呼吸试验结果的相关性分析
质酶-3样蛋白1(CHI3L1),是Th2型炎症和组织重塑的关键调节因子[7]㊂Bouvet等[8]证明,在小鼠模型中,CHI3L1有助于Th2型炎症和肺纤维化的发生,并抑制Th1型炎症㊂临床证据表明,COPD病例肺组织中YKL-40表达水平升高[9]㊂然而,YKL-40在COPD 患者中的作用仍不清楚㊂因此,本研究在COPD病例中检测血清YKL-40水平,并发现与对照组相比, COPD患者血清YKL-40水平升高,并且在COPD病例中,随着患者肺功能的下降,YKL-40水平逐渐升高㊂此外,相关性分析显示,COPD患者血清YKL-40水平与IL-6㊁TNF-α㊁CRP呈正相关㊂因此,YKL-40可能参与COPD的进展㊂在本研究中,回归分析结果证实血清YKL-40水平与FEV1%呈负相关,和与住院时间ȡ13d呈正相关,表明入院时较高的血清YKL-40水平反应了COPD的严重程度㊂CXCL9是一种IFN-γ诱导的趋化因子,由嗜中性粒细胞释放,并作为呼吸系统疾病中的Th1型炎症标记物㊂与健康对照组相比,COPD患者的CXCL9浓度增加,并与嗜中性气道炎症标记物呈正相关㊂Fulker-son等证明,CXCL9抑制嗜酸性粒细胞迁移至过敏原诱导小鼠的肺部炎症[10]㊂总之,这些发现表明CXCL9与抑制的Th2型炎症和增加的Th1型炎症相关㊂在这项研究中,我们没有发现CXCL9水平在COPD患者和健康对照之间的明显差异,这与其他研究报道的结果不一致㊂这些发现可能是由于我们的样本量太小,在分析中没有统计学意义㊂因此,需要更多参与者的进一步研究来阐明血清CXCL9水平和COPD患者严重程度㊁预后之间的关系㊂综上所述,COPD患者血清YKL-40水平升高与肺功能下降和住院时间延长相关,表明YKL-40可能参与了COPD的发病机制㊂因此,入院时YKL-40定量有望补充基于炎症生物标志物的COPD严重程度评估㊂ʌ参考文献ɔ[1]㊀张炜,丁朵,王蕾.慢性阻塞性肺疾病相关肺动脉高压中肺动脉压力与肺功能的关联性研究[J].西安交通大学学报:医学版,2022,43(6):889-894.[2]㊀陈训春,李名兰,潘碧云,等.TLR4/NF-κB信号通路激活LncRNA RP11-20G6调控慢性阻塞性肺疾病气道炎症和重塑[J].安徽医科大学学报,2022,57(4):586-593. [3]㊀Singh D,Bafadhel M,Brightling C E,et al.Blood eosinophilcounts in clinical trials for chronic obstructive pulmonary dis-ease[J].Am Respir Crit Care Med,2020,202(5):660-671.[4]㊀Jeong M H,Han H,Lagares D,et al.Recent advances in mo-lecular diagnosis of pulmonary fibrosis for precision medicine[J].ACS Pharmacol Transl Sci,2022,5(8):520-538.[5]㊀Coriati A,Bouvet G F,Masse C,et al.Ykl-40as a clinicalbiomarker in adult patients with cf:Implications of a chi3l1single nucleotide polymorphism in disease severity[J].CystFibros,2021,20(6):e93-e99.[6]㊀Jiang Y L,Fei J,Cao P,et al.Serum cadmium positively cor-relates with inflammatory cytokines in patients with chronicobstructive pulmonary disease[J].Environ Toxicol,2022,37(1):151-160.[7]㊀Kamle S,Ma B,He C H,et al.Chitinase3-like-1is a thera-peutic target that mediates the effects of aging in COVID-19[J].JCI insight,2021,6(21):148749.[8]㊀Bouvet G F,Bulka O,Coriati A,et al.Peripheral bloodmononuclear cell response to YKL-40and Galectin-3incystic fibrosis[J].Cytokine,2021,146:155635. [9]㊀Hakansson K E J,Ulrik C S,Godtfredsen N S,et al.High su-PAR and low blood eosinophil count are risk factors for hos-pital readmission and mortality in patients with COPD[J].Int Chron Obstruct Pulmon Dis,2020,15:733-743. [10]㊀Hasegawa T,Okazawa T,Uga H,et al.Serum CXCL9as apotential marker of Type1inflammation in the context of e-osinophilic asthma[J].Allergy,2019,74(12):2515-2518.ʌ文章编号ɔ1006-6233(2023)12-2059-05幽门螺杆菌感染内镜下胃黏膜表现与14C尿素呼吸试验结果的相关性分析蔡㊀雯,㊀赵㊀玲,㊀吴学勇,㊀涂㊀丽(皖北煤电集团总医院消化内科,㊀安徽㊀宿州㊀234000)ʌ摘㊀要ɔ目的:探究幽门螺杆菌(Hp)感染内镜下胃黏膜表现与14C尿素呼吸试验结果的相关性㊂9502ʌ基金项目ɔ2021年度宿州市科技计划项目,(编号:SZSKJJZC050)方法:以2021年9月至2022年9月102例慢性胃炎患者为研究对象,均给予内镜检查及14C尿素呼吸试验,比较Hp阳性组及Hp阴性组患者的14C-UBT检测值,并分析慢性胃炎患者内镜下表现与14C-UBT 检测值的关系;比较不同胃炎类型患者14C-UBT检测值㊂结果:Hp感染阳性患者14C-UBT检测值高于Hp感染阴性患者(P<0.05)㊂内镜下有皱襞增生的慢性胃炎患者14C-UBT检测值大于无皱襞增生,有萎缩患者的14C-UBT检测值大于无萎缩患者(P<0.05)㊂102例慢性胃炎患者中共检出浅表性胃炎40例㊁糜烂性胃炎9例㊁萎缩性胃炎53例,糜烂性胃炎㊁萎缩性胃炎患者的14C-UBT检测值大于浅表性胃炎患者(P<0.05)㊂结论:慢性胃炎患者14C-UBT检测值与Hp感染及内镜下表现有关,且胃炎类型也与14C-UBT检测值有关㊂ʌ关键词ɔ㊀慢性胃炎;㊀幽门螺旋杆菌;㊀内镜表现;㊀14C尿素呼吸试验ʌ文献标识码ɔ㊀A㊀㊀㊀㊀㊀ʌdoiɔ10.3969/j.issn.1006-6233.2023.12.023Correlation Analysis between Endoscopic Gastric Mucosal Manifestations of Helicobacter Pylori infection and14C Urea Breath Test ResultCAI Wen,ZHAO Ling,WU Xueyong,et al(Wanbei Coal and Electric Group General Hospital,Anhui Suzhou234000,China)ʌAbstractɔObjective:To explore the correlation between endoscopic gastric mucosal manifestations of Helicobacter pylori infection and14C urea breath test result.Methods:A total of102patients with chronic gastritis from September2021to September2022were selected as the study subjects.Endoscopic examination and14C urea breath test were performed,and14C-UBT detection value was compared between Hp-positive group and Hp-negative group and the relationship between endoscopic manifestations of patients with chronic gastritis and14C-UBT detection value was analyzed.14C-UBT detection value was compared among patients with different types of gastritis.Results:The14C-UBT detection value of patients with positive Hp infection was higher than that of patients with negative Hp infection(P<0.05).The14C-UBT detection value of chron-ic gastritis patients with endoscopic plicae hyperplasia was greater than that of patients without plicae hyperpla-sia.The14C-UBT detection value of patients with atrophy was greater than that of patients without atrophy(P <0.05).40cases of superficial gastritis,9cases of erosive gastritis and53cases of atrophic gastritis were de-tected among102patients with chronic gastritis.The14C-UBT detection value in patients with erosive gastritis or atrophic gastritis was greater than that of patients with superficial gastritis(P<0.05).Conclusion:The14C -UBT detection value of patients with chronic gastritis is correlated with Hp infection and endoscopic manifes-tations,and the type of gastritis is also related to14C-UBT detection value.ʌKey wordsɔ㊀Chronic gastritis;㊀Helicobacter pylori;㊀Endoscopic manifestations;㊀14C Urea breath test㊀㊀幽门螺旋杆菌(Helicobacter pylori,Hp)是一种革兰氏阴性菌,常寄存于胃窦及十二指肠,可导致患者出现慢性胃炎㊁消化性溃疡等疾病,且与胃癌及胃黏膜相关疾病有着密切关联[1]㊂既往报道指出,Hp可引起胃黏膜慢性活动性炎症,使得胃黏膜从正常状态转变为慢性非萎缩性胃炎,经进一步进展可发展为萎缩性胃炎等,最终进展为胃癌,提示Hp感染与慢性胃炎病情进展有关[2]㊂目临床上采用14C尿素呼吸试验㊁快速尿素酶试验等对Hp感染情况进行诊断,既往报道指出,14C尿素呼吸试验对Hp感染具有较高的诊断价值[3],但目前对于14C尿素呼吸试验结果与Hp感染程度的关系仍处于探索阶段,且其与慢性胃炎内镜下表现尚不明确㊂故本研究旨在探究幽门螺杆菌(Hp)感染内镜下胃黏膜表现与14C尿素呼吸试验的关系,为分析幽门螺旋菌感染于内镜下表现的关系提供参考依据㊂1㊀资料与方法1.1㊀临床资料:以2021年9月至2022年9月102例慢性胃炎门诊患者为研究对象,其中男29例,女73例;年龄13~69岁,平均(38.72ʃ12.47)岁㊂1.2㊀纳入标准:①符合有关慢性胃炎的诊断标准(内镜下可见黏膜红斑或出血点,或表现为黏膜红白相0602间)[4];②患者已签署同意书;③均行内镜检查及14C 尿素呼吸试验㊂1.3㊀排除标准:①疾病危重期患者;②合并消化道结构异常者;③既往有消化道手术史者;④合并消化道肿瘤患者;⑤因疾病不能耐受检查者;⑥精神疾病患者㊂1.4㊀方㊀法1.4.1㊀Hp感染诊断标准:参照相关指南对患者Hp 感染进行判定,即胃黏膜组织病理检查阳性或HpSA 检测阳性(患者入院24h内)[5]㊂1.4.2㊀14C尿素呼吸试验:患者于禁食12h后服用14C 尿素胶囊(中核海得威生物科技),以20mL温开水送服(不可咀嚼),20min后进行分析,用一根长约20cm 带滴球的一次性输液管向闪烁瓶(1mL氢氧化胺+1mL 无水乙醇+1滴1%酚酞)中吹气,应注意力度适中,放置液体喷出,当CO2吸收剂的颜色(紫红色)褪去时,提示CO2饱和,停止吹气㊂取5mL闪烁瓶中液体,应用LKB1217型液闪计数仪测定每分钟衰变数,以检测值>99dpm为阳性㊂1.4.3㊀内镜表现:应用日本奥林巴斯公司CLV-290SL290胃镜进行检查,参照相关指南对不同类型慢性胃炎进行分型㊂浅表性胃炎:Ⅰ级:间断线状红斑;Ⅱ级:有密集红斑点;Ⅲ级:广泛融合红斑㊂糜烂性胃炎:Ⅰ级:单发糜烂;Ⅱ级:糜烂处ɤ5处;Ⅲ级:糜烂处ȡ6处㊂萎缩性胃炎:Ⅰ级:黏膜有细颗粒,可见部分血管;Ⅱ级:黏膜有中等颗粒,可见连续均匀血管;Ⅲ级:黏膜有粗大颗粒且皱襞消失[6]㊂1.5㊀统计学处理:研究所得数据均用SPSS17软件处理,计数资料以百分比表示,采用χ2检验比较组间差异;计量资料经正态检验后用( xʃs)表示,用独立样本t检验比较组间差异,多组间比较采用单因素方差分析,多组均数间两两比较采用Newman-kueuls法㊂P< 0.05即差异具有统计学意义㊂2㊀结㊀果2.1㊀Hp感染阳性及阴性患者的14C-UBT检测值比较:感染阳性患者14C-UBT检测值高于Hp感染阴性患者(P<0.05),见表1㊂表1㊀Hp感染阳性及阴性患者的14C-UBT检测值比较(dpm)分组例数14C-UBT检测值Hp感染阳性81435.05ʃ50.38 Hp感染阴性2176.16ʃ19.74 t31.918 P<0.0012.2㊀不同内镜下表现与14C-UBT检测值的关系:内镜下有皱襞增生的慢性胃炎患者14C-UBT检测值大于无皱襞增生,有萎缩患者的14C-UBT检测值大于无萎缩患者(P<0.05),见表2㊂表2㊀内镜下表现与14C-UBT检测值的关系(dpm)内镜下表现例数14C-UBT检测值t P规则排列集合静脉有37378.03ʃ62.18 1.8530.067无65351.56ʃ73.09线状红斑有31375.59ʃ68.72 1.4860.141无71354.86ʃ63.07增生性息肉有25371.02ʃ69.310.8350.406无77357.96ʃ67.54皱襞增生有65394.02ʃ61.857.312<0.0011602无37303.43ʃ57.03黄色瘤有25371.64ʃ63.02 1.0090.316无77357.76ʃ58.71萎缩有53421.57ʃ67.9210.427<0.001无49295.82ʃ52.132.3㊀不同疾病类型患者14C-UBT检测值比较:102例慢性胃炎患者中共检出浅表性胃炎40例㊁糜烂性胃炎9例㊁萎缩性胃炎53例,糜烂性胃炎㊁萎缩性胃炎患者的14C-UBT检测值大于浅表性胃炎患者(P<0.05),见表3㊂表3㊀不同疾病类型患者14C-UBT检测值比较(dpm)分组例数14C-UBT检测值浅表性胃炎40331.45ʃ42.79糜烂性胃炎9381.71ʃ46.35∗萎缩性胃炎53380.09ʃ50.31∗F13.060 P<0.001㊀㊀注:与浅表性胃炎比较,∗P<0.053㊀讨㊀论Hp是一类显微镜下呈螺旋形或弧形的需氧细菌,具有较高的传染性,感染患者多无明显症状,少数患者或会出现食欲下降㊁腹痛等症状㊂据相关报道显示,远超半数中老年人感染过Hp[7]㊂胃黏膜组织快速尿素酶试验是检查Hp感染的主要手段,是通过电子胃镜取胃黏膜或组织进行快速尿素酶试验,来对Hp感染情况进行诊断,虽具有较高的准确率,但其为有创操作,且过程较为痛苦,导致重复性较低㊂相关报道指出,14C尿素呼吸试验对Hp感染也具有较好的诊断价值,且具有无创等优点[8]㊂但有研究显示,无法仅通过14C尿素呼吸试验对Hp进行诊断[9]㊂受试者如胃内存在Hp,当其在服用一定量的14C尿素后,示踪尿素可被Hp产生的尿素酶分解,以14CO2的形式经肺排出,最终被仪器检查出呼出气中14CO2含量㊂本研究发现,Hp感染患者的14C-UBT检测值均高于未感染患者,与相关研究结果相符[10],提示该方法可对Hp感染进行诊断㊂经进一步分析发现,患者14C-UBT检测值与皱襞增生㊁萎缩等内镜下表现有关,表明该内镜下表现与Hp感染相关,提示或可通过观察该内镜下表现来对患者是否存在Hp感染进行初步判断㊂Hp感染主要与胃炎㊁消化道溃疡及萎缩性胃炎等胃肠疾病关系密切,Hp在感染机体后可导致中性粒细胞㊁单核细胞浸润,使胃黏膜毛细血管网充血扩张,进而导致慢性胃炎的发生[11]㊂既往报道指出,Hp感染与胃炎严重程度有关[12]㊂慢性胃炎的发展过程中,持续Hp感染促进胃黏膜向萎缩性㊁糜烂性方向发展㊂本研究结果显示,不同类型胃炎患者14C-UBT检测值对比存在显著性差异,提示患者Hp感染与疾病类型有关㊂其原因在于,Hp持续感染可在炎症作用下使胃黏膜出现持续损伤,并且Hp定植于胃黏膜上皮细胞表面和胃黏液底层,破坏了胃黏膜屏障,使得胃黏膜更易受到细菌损伤,进而可加重胃黏膜损伤程度,促进胃黏膜向萎缩性㊁糜烂性方向进展[12,13]㊂综上所述,慢性胃炎患者14C-UBT检测值与Hp 感染及内镜下表现有关,且胃炎类型也与14C-UBT检测值有关㊂但本研究尚存在不足之处,患者纳入样本量较少可能会导致研究结果出现偏倚,故后期需增加样本量以进行进一步分析㊂ʌ参考文献ɔ[1]㊀雷蓉,杨丹,袁芳桃,等.10661例体检者幽门螺杆菌感染情况及其相关危险因素分析[J].基础医学与临床,2022, 42(1):126-130.[2]㊀吴正奇,张志镒,陶雪梅,等.甘肃省某医院14C呼气试验受检者幽门螺杆菌感染现状[J].兰州大学学报:医学版, 2022,48(4):56-58.[3]㊀Beresniak A,Malfertheiner P,Franceschi F,et al.Helico-bacter pylori"Test-and-Treat"strategy with urea breathtest:A cost-effective strategy for the management of dyspep-sia and the prevention of ulcer and gastric cancer in SpainResults of the Hp-Breath initiative[J].Helicobacter,2020,25(4):12693.[4]㊀房静远,杜奕奇,刘文忠,等.中国慢性胃炎共识意见(2017年,上海)[J].中华消化杂志,2017,37(11):721-738.[5]㊀中华医学会,中华医学会杂志社,中华医学会全科医学分2602会,等.幽门螺杆菌感染基层诊疗指南(实践版㊃2019)[J ].中华全科医师杂志,2020,19(5):403-407.[6]㊀中华消化内镜杂志中华医学会消化内镜学分会.慢性胃炎的内镜分型分级标准及治疗的试行意见[J ].中华消化内镜杂志,2004,21(2):77-78.[7]㊀耿甜,李中跃.13C -尿素呼气试验诊断儿童幽门螺杆菌感染相关影响因素[J ].中华实用儿科临床杂志,2022,37(7):552-555.[8]㊀Yan L ,Chen Y ,Chen F ,et al.Effect of helicobacter pylori e-radication on gastric cancer prevention :updated report from a randomized controlled trial with 26.5years of follow -up [J ].Gastroenterology ,2022,163(1):154-162.[9]㊀杨志平,田婉佳,王远志,等.口腔幽门螺杆菌感染与幽门螺杆菌性胃炎的关系[J ].北华大学学报(自然科学版),2022,23(3):352-356.[10]㊀Bhat S ,Nunes D.Pharmacist -managed helicobacter pyloritreatment service within a gastroenterology clinic :workflowand real -world experiences :[J ].Ann Pharmacother ,2022,56(2):162-169.[11]㊀贾纯增,陈锐,王玘,等.13C -尿素呼气试验数值与幽门螺旋杆菌根除率相关性研究[J ].临床军医杂志,2021,49(4):403-404.[12]㊀杨洋,高广周,郝英霞.白光胃镜下内镜征象诊断幽门螺杆菌感染的临床研究[J ].河北医科大学学报,2022,43(4):391-396.[13]㊀刘庆华,李真,张晓伟,等.胃镜下病理改变与血清幽门螺杆菌抗体分型的关系分析[J ].内科理论与实践,2022,17(4):313-316.ʌ文章编号ɔ1006-6233(2023)12-2063-06椎体CT 值与经皮椎体成形术骨水泥渗漏的相关性研究高志超,㊀李㊀哲,㊀许财元,㊀张仁赞,㊀王建华,㊀孙㊀贺(承德医学院附属医院脊柱外科,㊀河北㊀承德㊀067000)ʌ摘㊀要ɔ目的:分析椎体CT 值与骨质疏松性椎体压缩性骨折(OVCF )的患者行经皮椎体成形术(PVP )治疗发生骨水泥渗漏是否具有一定相关性㊂方法:收集承德医学院附属医院在2019年9月至2022年10月之间因骨质疏松性椎体压缩骨折住院并且行PVP 治疗的患者61例,将所有椎体按照是否发生骨水泥渗漏分为渗漏组和未渗漏组,并记录两组患者的性别㊁年龄㊁术中骨水泥的注入量,骨折部位(胸椎或腰椎)㊁骨折压缩程度及腰1椎体CT 值㊂结果:两组的性别㊁年龄㊁骨折部位(胸椎或腰椎)及骨水泥注入量的变化差异无统计学意义(P >0.05),骨折压缩程度㊁腰1椎体的CT 值差异有统计学意义(P <0.05),将两者纳入多因素二元Logistic 分析后得出腰1椎体CT 值的变化与骨水泥渗漏的关联有统计学意义(P <0.05),进一步做ROC 曲线分析,曲线下的面积(AUC )为0.674(95%CI :0.524~0.824,P =0.025),得出腰1椎体CT 值的最佳截断值为62.1HU 时,其敏感度为79.5%,特异度为63.6%㊂结论:腰1椎体CT 值与PVP 治疗骨质疏松性椎体压缩骨折骨水泥渗漏具有相关性,在术前测量患者的腰1椎体CT 值在评估骨水泥渗漏方面有一定的指导意义㊂ʌ关键词ɔ㊀CT 值;㊀经皮椎体成形术;㊀骨水泥渗漏;㊀骨质疏松;㊀椎体压缩骨折ʌ文献标识码ɔ㊀A㊀㊀㊀㊀㊀ʌdoi ɔ10.3969/j.issn.1006-6233.2023.12.024Correlation Study between Vertebral CT Values and Bone CementLeakage in Percutaneous Vertebral Augmentation forOsteoporotic Vertebral Compression FracturesGAO Zhichao ,LI Zhe ,XU Caiyuan ,et al(Affiliated Hospital of Chengde Medical University ,HebeiChengde 067000,China )ʌAbstract ɔObjective :To analyze the correlation between vertebral CT values and the occurrence of3602ʌ基金项目ɔ河北省承德市科学技术研究与发展计划项目,(编号:201904A038)ʌ通讯作者ɔ李㊀哲。
医学术语谷氨酸脱氢酶英文
医学术语谷氨酸脱氢酶英文Title: Glutamate Dehydrogenase: A Key Enzyme in Amino Acid Metabolism.Glutamate Dehydrogenase (GDH) is an enzyme that plays a crucial role in amino acid metabolism. It catalyzes the reversible conversion of glutamate to alpha-ketoglutarate, a process that involves the oxidation of glutamate and the reduction of NAD+ to NADH+H+. This enzyme is found in various tissues of the body, including the liver, kidney, and brain, and it plays a vital role in maintaining the balance of amino acids in the body.The structure of GDH is complex, with multiple subunits that come together to form the active enzyme. The enzyme requires NAD+ or NADP+ as a cofactor for its activity, and it can exist in either the mitochondrial or cytosolic form, depending on the tissue and species.The function of GDH is diverse and depends on itssubcellular location. In the mitochondria, GDH plays a role in the Krebs cycle, where it converts glutamate to alpha-ketoglutarate, generating NADH+H+ in the process. This NADH+H+ can then be used by the electron transport chain to generate ATP, the cell's main energy currency. In the cytosol, GDH plays a role in amino acid catabolism, whereit converts glutamate to alpha-ketoglutarate, generating NADH+H+ in the process. This NADH+H+ can then be used by other enzymes in the cytosol for various biosynthetic reactions.The regulation of GDH activity is complex and involves multiple mechanisms. One of the main regulators of GDH activity is the availability of its substrates and cofactors. For example, an increase in the concentration of glutamate or NAD+ can stimulate GDH activity, while a decrease in these substrates can inhibit the enzyme. Additionally, GDH activity can be regulated by allosteric effectors, which bind to the enzyme and modify its activity. Some allosteric effectors, such as ADP and ATP, can inhibit GDH activity, while others, such as GTP, can stimulate the enzyme.The role of GDH in disease is also well-documented. Alterations in GDH activity have been observed in various diseases, including liver disease, kidney disease, and neurological disorders. For example, in liver disease, a decrease in GDH activity can lead to a decrease in the conversion of glutamate to alpha-ketoglutarate, resultingin an accumulation of glutamate in the blood. This accumulation can lead to neurological symptoms such as seizures and coma. Similarly, in kidney disease, a decrease in GDH activity can lead to an accumulation of glutamate in the urine, which can contribute to the development ofkidney stones.In conclusion, Glutamate Dehydrogenase is a key enzymein amino acid metabolism that plays a crucial role in maintaining the balance of amino acids in the body. Its function is diverse and depends on its subcellular location, and its activity is regulated by multiple mechanisms. Alterations in GDH activity have been observed in various diseases, indicating its importance in maintaining thehealth of the body. Future research on GDH may lead to abetter understanding of its role in disease and the development of new therapeutic strategies.。
Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes(NI
Sibirny 169, Elaine C.M. Silva-Zacarin 170, Hans-Uwe Simon 171, Cristiano Simone 172, Anne Simonsen 173, Mark A. Smith 174, Katharina Spanel-Borowski 175, Vickram Srinivas 168,Meredith Steeves 34, Harald Stenmark 173, Per E. Stromhaug 176, Carlos S. Subauste 177,Seiichiro Sugimoto 178, David Sulzer 179, Toshihiko Suzuki 180, Michele S. Swanson 181, Ira Tabas 182, Fumihiko Takeshita 183, Nicholas J. Talbot 184, Zsolt Tallóczy 179, KeijiTanaka 95, Kozo Tanaka 185, Isei Tanida 186, Graham S. Taylor 187, J. Paul Taylor 188, Alexei Terman 189, Gianluca Tettamanti 190, Craig B. Thompson 102, Michael Thumm 191, Aviva M.Tolkovsky 192, Sharon A. Tooze 193, Ray Truant 194, Lesya V. Tumanovska 195, YasuoUchiyama 196, Takashi Ueno 96, Néstor L. Uzcátegui 197, Ida van der Klei 89, Eva C.Vaquero 198, Tibor Vellai 199, Michael W. Vogel 200, Hong-Gang Wang 201, Paul Webster 202,John W. Wiley 203, Zhijun Xi 204, Gutian Xiao 205, Joachim Yahalom 206, Jin-Ming Yang 207,George Yap 208, Xiao-Ming Yin 209, Tamotsu Yoshimori 139, Li Yu 107, Zhenyu Yue 210,Michisuke Yuzaki 211, Olga Zabirnyk 212, Xiaoxiang Zheng 213, Xiongwei Zhu 174, and Russell L. Deter 2141 Life Sciences Institute, and Departments of Molecular, Cellular and Developmental Biology and Biological Chemistry; University of Michigan; Ann Arbor, Michigan USA2 Department of Biochemistry and Food Science; Hebrew University; Rehovot, Israel3 Department of Molecular Cell Biology; Catholic University of Leuven; Leuven, Belgium4 Creighton University School of Medicine; Department of Biomedical Sciences;Omaha, Nebraska USA5 Department of Biology; College of Sciences; University of Texas at San Antonio;San Antonio, Texas USA6 Department of Pathology & Laboratory Medicine; University of Cincinnati College of Medicine; Cincinnati, Ohio USA7 Department of Chemical and Biological Sciences; Japan Women’s University; Tokyo, Japan8 Department of Cancer Biology; University of Massachusetts Medical School;Worcester, Massachusetts USA9 Department of Pharmaceutical Sciences; University of Connecticut; Storrs,Connecticut USA 10 Telethon Institute of Genetics and Medicine; Napoli, Italy 11 Department of Biological Sciences; University of Toledo; Toledo, Ohio USA 12 Department of Genetics, Development and Cell Biology,and Plant Sciences Institute; Iowa State University; Ames, Iowa USA 13 Center for Research on Biology and Pathology of Aging; University of Pisa; Pisa, Italy 14 Basic Medical Sciences; Western University of Health Sciences; Pomona, California USA 15 Centre d’études d’agents Pathogènes et Biotechnologies pour la Santé;CNRS UM1, UM2; Institut de Biologie; Montpellier, France 16 Department of Microbiology andImmunology; Indiana University School of Medicine; Indianapolis, Indiana USA 17 Buck Institute for Age Research; Novato, California USA 18 Department of Biological Sciences; University of Pittsburgh;Pittsburgh, Pennsylvania USA 19 Cell Biology Program; Hospital for Sick Children; Toronto, Ontario,Canada 20 Division of Pharmacology; Faculty of Health Sciences; Linköping University; Linköping, Sweden 21 Department of Medicine I; Division of Oncology; Institute of Cancer Research; Medical University of Vienna; Vienna, Austria 22 UMR5095; CNRS; Université de Bordeaux 2; Bordeaux, France 23 Department of Microbiology; Instituto de Fermentaciones Industriales; Madrid, Spain 24 Dulbecco Telethon Institute—IRCCS Santa Lucia Foundation and Department of Biology; University of Rome Tor Vergata; Rome, Italy 25 Department of Immunology; Peking University; Center for Human Disease Genomics; Beijing, China 26 Department of Pharmacology; Emory University School of Medicine; Atlanta, Georgia USA 27 Division of Pulmonary and Critical Care Medicine; Brigham & Womens Hospital; Harvard Medical School; Boston,Massachusetts USA 28 Department of Pathology and Center for Neuroscience; University of Pittsburgh;Pittsburgh, Pennsylvania USA 29 National Creative Research Initiatives Center for Cell Growth Regulation;Department of Biological Sciences; Korea Advanced Institute of Science and Technology; Republic of Korea 30 Département de Biologie Cellulaire et de Morphologie; Université de Lausanne; Lausanne, Switzerland 31 Safar Center for Resuscitation Research; Pittsburgh, Pennsylvania USA 32 Department of Chemistry and Biochemistry and the Molecular Biology Institute; University of California, Los Angeles; Los Angeles,California USA 33 Laboratoire de Génétique Moléculaire des Champignons; Institut de Biochimie et de Génétique Cellulaires; CNRS; Université de Bordeaux 2; Bordeaux, France 34 Department of CancerBiology; The Scripps Research Institute; Jupiter, Florida USA 35 INSERM U756, and the Université Paris-Sud 11; Châtenay-Malabry, France 36 Laboratorio de Biología Celular y Molecular-Instituto de Histología NIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author Manuscripty Embriología; Universidad Nacional de Cuyo-CONICET; Mendoza, Argentina 37 Departamento deMorfología y Biología Celular; Universidad de Oviedo; Oviedo, Spain 38 Keck Graduate Institute of Applied Sciences; Claremont, California USA 39 Department of Anatomy and Structural Biology and ofDevelopmental and Molecular Biology; Marion Bessin Liver Research Center; Albert Einstein College of Medicine; Bronx, New York USA 40 Department of Pathology; University of California, San Francisco; San Francisco, California USA 41 Laboratorio Nazionale Consorzio Interuniversitario Biotecnologie; Trieste,Italy 42 Department of Genome Science; University of Cincinnati; Cincinnati, Ohio USA 43 Medical Oncology Branch; National Cancer Institute/Navy Medical Oncology; Bethesda, Maryland USA 44Department of Molecular Genetics and Microbiology; University of New Mexico Health Science Center;Albuquerque, New Mexico USA 45 Department of Biochemistry & Molecular Biology; and ARC Centre of Excellence in Structural and Functional Microbial Genomics; Monash University; Clayton Campus;Melbourne, Victoria, Australia 46 Department of Biology; University of Tor Vergata; via della Ricerca Scientifica; Rome, Italy 47 Department of Physiology; Tufts University; Boston, Massachusetts USA 48Department of Neurology; Massachusetts General Hospital; Charlestown, Massachusetts USA 49Department of Molecular, Cellular & Developmental Biology; Yale University; New Haven, Connecticut 50 Departments of Medicine, Pharmacology and Pathology; Comprehensive Cancer Center; Case Western Reserve University and University Hospitals of Cleveland; Cleveland, Ohio USA 51 Immunotec Research Ltd.; Quebec, Canada 52 Virologie et Immunologie Moleculaires; INRA UR892; Jouy-en-Josas, France 53 Department of Anatomy and Cell Biology; University of Florida College of Medicine; Gainesville, Florida USA 54 Department of Biochemistry; University of Tuebingen; Tuebingen, Germany 55 Baylor College of Medicine; Houston, Texas USA 56 Department of Biological Chemistry; The Weizmann Institute of Science;Rehovot, Israel 57 Department of Biological and Environmental Sciences; University of Helsinki; Helsinki,Finland 58 Departments of Biochemistry and Molecular Biology; Apoptosis and Genomics Research Group of the Hungarian Academy of Sciences; Research Center for Molecular Medicine; University of Debrecen;Debrecen, Hungary 59 Cellular Neurobiology Laboratory; The Salk Institute for Biological Studies; San Diego, California USA 60 Centro de Investigación Biomédica en Red de Enfermedades Neurodegenerativas (CIBERNED); Departamento Bioquímica y Biología Molecular y Genética; E.U. Enfermería; Universidad de Extremadura; Cáceres, Spain 61 Department of Neuro-Oncology; MD Anderson Cancer Center;University of Texas; Houston, Texas USA 62 Department of Frontier Veterinary Medicine; KagoshimaUniversity; Kagoshima, Japan 63 Department of Zoology and Animal Biology; University of Geneva; Geneva,Switzerland 64 Gladstone Institute of Neurological Disease and Department of Neurology; University of California; San Francisco, California USA 65 Department of Pharmacology and Toxicology and Massey Cancer Center; Virginia Commonwealth University; Richmond, Virginia USA 66 Biochemistry and Medical Genetics; Manitoba Institute of Cell Biology; Winnipeg, Manitoba, Canada 67 Institute for Biochemistry and Molecular Biology II; Heinrich Heine University; Düsseldorf, Germany 68 Department of Cell Biology;Harvard Medical School; Boston, Massachusetts USA 69 Department of Developmental Genetics and Gene Control; Institute of Genetics; The University of Nottingham; Queen’s Medical Centre; Nottingham UK 70Genome Sciences Center; British Columbia Cancer Agency; Vancouver, British Columbia, Canada 71BioScience Center; San Diego State University, San Diego; San Diego, California USA 72 Deparment of Internal Medicine; Heinrich Heine Universität; Düsseldorf, Germany 73 Department of Immunology; Duke University Medical Center; Durham, North Carolina USA 74 Department of Pharmacology; University of Colorado Health Sciences Center; Aurora, Colorado USA 75 Division of Cardiology; University of Texas Southwestern Medical Center; Dallas, Texas USA 76 Apoptosis Department and Centre for Genotoxic Stress Research; Institute of Cancer Biology; Danish Cancer Society; Copenhagen, Denmark 77 Cancer Institute,The Second Affiliated Hospital; Zhejiang University School of Medicine; Hangzhou, Zhejiang, China 78Department of Life Science; National Taiwan University; Taipei, Taiwan 79 Department of Immunobiology;Yale University School of Medicine; New Haven, Connecticut USA 80 Department of Microbiology and Molecular Genetics; Medical College of Wisconsin; Milwaukee, Wisconsin USA 81 Memorial Sloan-Kettering Cancer Center; New York, New York USA 82 Department of Pharmacology; University of Medicine and Dentistry of New Jersey—Robert Wood Johnson Medical School; Piscataway, New Jersey USA 83NIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author ManuscriptBiochemistry Department; Institute of Medical Biology; University of Tromsø; Tromsø, Norway 84Department of Molecular Microbiology and Immunology; University of Southern California Keck Medical School; Los Angeles, California USA 85 Department of Applied Biological Chemistry; Niigata University;Niigata, Japan 86 Department of Molecular Biology; University of Texas Southwestern Medical Center;Dallas, Texas USA 87 Department of Laboratory Medicine and Pathology; University of Minnesota Cancer Center; Minneapolis, Minnesota USA 88 Department of Medicine & Pharmacology; Wayne State University School of Medicine; Detroit, Michigan USA 89 Molecular Cell Biology; Groningen Biomolecular Sciences and Biotechnology Institute (GBB); University of Groningen; Haren, The Netherlands 90 Division of Pulmonary, Allergy and Critical Care Medicine; University of Pittsburgh Medical Center; Pittsburgh,Pennsylvania USA 91 Department of Molecular Genetics; Weizmann Institute of Science; Rehovot, Israel 92 Department of Radiation Oncology; University Hospitals of Cleveland; Cleveland, Ohio USA 93Department of Biotechnology; University of Tokyo; Tokyo, Japan 94 Department of Cell Biology; Centro de Investigación Príncipe Felipe; Valencia, Spain 95 Laboratory of Frontier Science; Tokyo Metropolitan Institute of Medical Science; Tokyo, Japan 96 Department of Biochemistry; Juntendo University School of Medicine; Tokyo, Japan 97 Department of Neurosurgery; The University of Texas MD Anderson Cancer Center; Houston, Texas USA 98 Department of Anatomy, Cell and Developmental Biology; Eötvös Loránd University; Budapest, Hungary 99 INSERM U848; Institut Gustave Roussy, and the Université Paris-Sud 11;Villejuif, France 100 Division of Developmental Biology; Cincinnati Children’s Hospital ResearchFoundation; Cincinnati, Ohio USA 101 Molecular and Cellular Oncology; MD Anderson Cancer Center;Houston, Texas USA 102 Abramson Family Cancer Research Institute; University of Pennsylvania School of Medicine; Philadelphia, Pennsylvania USA 103 Laval University Cancer Research Center; Québec,Canada 104 Department of Biology; Eastern Michigan University; Ypsilanti, Michigan USA 105 Institute of Health Sciences; Shanghai Jiao Tong University School of Medicine & Shanghai Institutes for Biological Sciences; Chinese Academy of Sciences; Shanghai, China 106 Department of Microbiology and Immunology;National Cheng Kung University; Taiwan 107 Molecular Development Section; Laboratory of Immunology;National Institute of Allergy and Infectious Diseases; National Institutes of Health; Bethesda, Maryland USA 108 Departments of Internal Medicine and Microbiology; University of Texas Southwestern Medical Center;Dallas, Texas USA 109 University of Michigan Medical School; Department of Pathology; Ann Arbor,Michigan USA 110 Neurodegeneration Research Laboratory; National Neuroscience Institute; Singapore 111 Biotechnology Institute; Zhejiang University; Hangzhou, China 112 Department of Anatomy; Chang Gung University; Taiwan 113 Department of Experimental Therapeutics; University of Texas MD Anderson Cancer Center; Houston, Texas USA 114 Departamento de Bioquímica y Biología Molecular; InstitutoUniversitario de Oncología; Universidad de Oviedo; Oviedo, Spain 115 Department of Radiation Oncology;Vanderbilt University; Nashville, Tennessee USA 116 Ben May Department for Cancer Research; University of Chicago; Gordon Center for Integrative Sciences; Chicago, Illinois USA 117 Drug Research andEvaluation; Istituto Superiore di Sanita; Rome, Italy 118 Department of Pharmacology; University of Antwerp; Wilrijk, Antwerp, Belgium 119 Faculty of Agriculture; Kyushu University; Fukuoka, Japan 120GSF—National Research Center for Environment and Health; Munich, Germany 121 Department of Medical Biochemistry; Academic Medical Center; Amsterdam, The Netherlands 122 Biology Department; Queens College; City University of New York; Flushing, New York USA 123 Research Unit for Tropical Diseases de Duve Institute and Laboratory of Biochemistry; Université Catholique de Louvain; Brussels, Belgium 124Department of Biological Chemistry; University of Padua; Padua, Italy 125 Department of Healthcare Sciences; The University College of Antwerp; Antwerp, Belgium 126 Department of Physiology and Cell Biology; Tokyo Medical and Dental University; Tokyo, Japan 127 INSERM ERI 21, and the Laboratoire de Pathologie Clinique et Expérimentale; IFR-50; Nice, France 128 Department of Cell Biology; University Medical Centre Utrecht; Utrecht, The Netherlands 129 Plymouth Marine Laboratory; Plymouth, UK 130Institute of Physiology; Center for Neuroscience and Cell Biology; Coimbra, Portugal 131 Division of Life Science; Graduate School of Science and Engineering; Saitama University; Saitama, Japan 132 Department of Physiological Sciences; Warsaw Agricultural University; Warsaw, Poland 133 Laboratory of ViralImmunobiology; The Rockefeller University; New York, New York USA 134 Novartis Institutes for BiomedicalNIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author ManuscriptResearch; Cambridge, Massachusetts USA 135 Fungal Patho-Biology Group; Temasek Life Sciences Laboratory; National University of Singapore; Singapore 136 Department of Genetics, Cell Biology &Development; University of Minnesota; Minneapolis, Minnesota USA 137 Department of Neuromuscular Research; National Institute of Neuroscience; National Center of Neurology and Psychiatry (NCNP);Kodaira, Tokyo, Japan 138 Nathan Kline Institute; New York University School of Medicine; Orangeburg,New York USA 139 Department of Cellular Regulation; Research Institute for Microbial Diseases; Osaka University; Osaka, Japan 140 Institute for Biochemistry and Molecular Biology II; Heinrich HeineUniversity, Düsseldorf; Düsseldorf, Germany 141 Department of Microbiology and Immunology; University of Tokyo; Tokyo, Japan 142 Departments of Radiation Oncology, Biochemistry and Environmental Health Sciences, and the Case Comprehensive Cancer Center; Case Western Reserve University; Cleveland, Ohio USA 143 Department of Molecular, Cellular and Developmental Biology; University of Michigan; Ann Arbor,Michigan USA 144 Department of Oncology; Chaim-Sheba Medical Center; Ramat-Gan, Israel 145Department of Oral and Craniofacial Biology; Louisiana State University Health Sciences Center School of Dentistry; New Orleans, Louisiana USA 146 Department of Cell Biology and Biophysics; Panepistimiopolis,Athens, Greece 147 Inflammatory Bowel Disease Research Group; Addenbrooke’s Hospital; University of Cambridge; Cambridge, UK 148 Department of Pediatrics; University of Pittsburgh School of Medicine and Children’s Hospital of Pittsburgh; Pittsburgh, Pennsylvania USA 149 Department of Biology; University of Tor Vergata; via della Ricerca Scientifica; Rome, Italy 150 Department of Neurobiochemistry; Tel-Aviv University; Ramat-Aviv, Tel-Aviv, Israel 151 Department of Biochemistry & Molecular Biology; Monash University; Clayton Campus; Melbourne, Victoria, Australia 152 Department of Molecular Biology;University of Tuebingen; Tuebingen, Germany 153 Arthritis and Rheumatism Branch; National Institute of Arthritis and Musculoskeletal and Skin Diseases; National Institutes of Health; Bethesda, Maryland USA 154 Institute of Cellular and Molecular Anatomy; Anatomie III; Clinic of the JWG-University; Frankfurt,Germany 155 Ophthalmology and Neurosciences Division; Medical University of South Carolina;Charleston, South Carolina USA 156 Department of Medical Genetics; Cambridge Institute for Medical Research; Cambridge, United Kingdom 157 Tumour Cell Death Laboratory; Beatson Institute for Cancer Research; Glasgow, Scotland UK 158 Department of Cell Biology and Molecular Medicine; Cardiovascular Research Institute; University of Medicine and Dentistry of New Jersey—New Jersey Medical School; Newark,New Jersey USA 159 Department of Diagnostic and Therapeutic Sciences; Meikai University School ofDentistry; Sakado, Japan 160 Division of Applied Life Sciences; Kyoto University; Kyoto, and JST; CREST;Kyoto, Japan 161 Department of Biomedical Science; University of Padova, Padova; and Dulbecco Telethon Institute at Venetian Institute of Molecular Medicine; Padova, Italy 162 Department of Microbiology and Immunology; University of Tokyo; Tokyo, Japan 163 Laboratorio Nazionale Consorzio Interuniversitario Biotecnologie; Trieste, Italy 164 Department of Cell Biology; Institute for Cancer Research; Rikshospitalet-Radiumhospitalet HF and Department of Molecular Biosciences; University of Oslo; Oslo, Norway 165Department of Animal Science; University of Wyoming; Laramie, Wyoming USA 166 Cancer Center;Massachusetts General Hospital; Charlestown, Massachusetts USA 167 Department of Pathology;University of Alabama at Birmingham; Birmingham, Alabama USA 168 Department of Orthopaedic Surgery;Thomas Jefferson University; Philadelphia, Pennsylvania USA 169 Institute of Cell Biology; NationalAcademy of Sciences of Ukraine; Lviv, Ukraine USA 170 Universidade Federal de Sao Carlos; Sorocaba,Brasil 171 Department of Pharmacology; University of Bern; Bern, Switzerland 172 Department ofTranslational Pharmacology; Consorzio Mario Negri Sud; Santa Maria Imbaro, Italy 173 Centre for Cancer Biomedicine; University of Oslo, and Department of Biochemistry; The Norwegian Radium Hospital;Montebello, Oslo, Norway 174 Department of Pathology; Case Western Reserve University; Cleveland, Ohio USA 175 Institute of Anatomy; University of Leipzig; Leipzig, Germany 176 Division of Biology; University of Missouri; Columbia, Missouri USA 177 Departments of Ophthalmology and Medicine; Case Western Reserve University School of Medicine; Institute of Pathology; Cleveland, Ohio USA 178 Department of Neurology; National Hospital Organization; Miyazaki Higashi Hospital; Miyazaki, Japan 179 Departments of Neurology and Psychiatry; Columbia University; New York, New York USA 180 Department ofMicrobiology; University of the Ryukyus; Okinawa, Japan 181 Department of Microbiology andNIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author ManuscriptImmunology; University of Michigan; Ann Arbor, Michigan USA 182 Department of Medicine; Columbia University; New York, New York USA 183 Department of Molecular Biodefense Research; Yokohama City University Graduate School of Medicine; Yokohama, Japan 184 School of Biosciences; University of Exeter;Exeter UK 185 Institute of Development, Aging and Cancer; Center for Research Strategy and Support;Tohoku University; Miyagi, Japan 186 Department of Biochemistry and Cell Biology; National Institute of Infectious Diseases; Tokyo, Japan 187 CRUK Institute for Cancer Studies; University of Birmingham;Birmingham, UK 188 Department of Neurology; University of Pennsylvania School of Medicine;Philadelphia, Pennsylvania USA 189 Division of Geriatric Medicine; Faculty of Health Sciences; Linköping University; Linköping, Sweden 190 Department of Structural and Functional Biology; University of Insubria;Varese, Italy 191 Zentrum Biochemie und Molekulare Zellbiologie; Georg-August-Universitaet; Goettingen,Germany 192 Department of Biochemistry; University of Cambridge; Cambridge UK 193 Secretory Pathways Laboratory; Cancer Research UK London Research Institute; London, England UK 194Biochemistry and Biomedical Sciences; McMaster University; Hamilton, Ontario, Canada 195 Department of General and Molecular Pathophysiology; Bogomoletz Institute of Physiology; Kiev, Ukraine 196Deptartment of Cell Biology and Neurosciences; Osaka University Graduate School of Medicine; Osaka,Japan 197 Escuela de Bioanálisis; Universidad Central de Venezuela; Caracas, Venezuela 198 Hospital Clínic; Division of Gastroenterology; Barcelona, Catalonia, Spain 199 Department of Genetics; Eötvös Loránd University; Budapest, Hungary 200 Maryland Psychiatric Research Center; Baltimore, Maryland USA 201 H. Lee Moffitt Cancer Center & Research Institute; Tampa, Florida USA 202 Ahmanson Center for Advanced Electron Microscopy & Imaging; House Ear Institute; Los Angeles, California USA 203Department of Internal Medicine; University of Michigan; Ann Arbor, Michigan USA 204 Department of Urology; Peking University First Hospital; Beijing, China 205 Department of Microbiology and Molecular Genetics; University of Pittsburgh School of Medicine; Pittsburgh, Pennsylvania USA 206 Department of Radiation Oncology; Memorial Sloan-Kettering Cancer Center; New York, New York USA 207 The Cancer Institute of New Jersey; University of Medicine and Dentistry of New Jersey—Robert Wood Johnson Medical School; New Brunswick, New Jersey USA 208 Department of Medicine; University of Medicine and Dentistry of New Jersey—New Jersey Medical School, Newark, New Jersey USA 209 Department of Pathology;University of Pittsburgh School of Medicine; Pittsburgh, Pennsylvania USA 210 Department of Neurology and of Neuroscience; Mount Sinai School of Medicine; New York, New York USA 211 Department of Neurophysiology; Keio University School of Medicine; Tokyo, Japan 212 Metabolism and CancerSusceptibility Section; Laboratory of Comparative Carcinogenesis; Center for Cancer Research; NCI—Frederick; National Institutes of Health; Frederick, Maryland USA 213 Department of BiomedicalEngineering; Zhejiang University; Zhejiang, China 214 Department of Obstetrics and Gynecology; Baylor College of Medicine; Houston, Texas USA AbstractResearch in autophagy continues to accelerate,1 and as a result many new scientists are entering thefield. Accordingly, it is important to establish a standard set of criteria for monitoringmacroautophagy in different organisms. Recent reviews have described the range of assays that havebeen used for this purpose.2,3 There are many useful and convenient methods that can be used tomonitor macroautophagy in yeast, but relatively few in other model systems, and there is muchconfusion regarding acceptable methods to measure macroautophagy in higher eukaryotes. A keypoint that needs to be emphasized is that there is a difference between measurements that monitorthe numbers of autophagosomes versus those that measure flux through the autophagy pathway; thus,a block in macroautophagy that results in autophagosome accumulation needs to be differentiatedfrom fully functional autophagy that includes delivery to, and degradation within, lysosomes (in mosthigher eukaryotes) or the vacuole (in plants and fungi). Here, we present a set of guidelines for theselection and interpretation of the methods that can be used by investigators who are attempting toexamine macroautophagy and related processes, as well as by reviewers who need to provide realisticand reasonable critiques of papers that investigate these processes. This set of guidelines is not meantNIH-PA Author Manuscript NIH-PA Author ManuscriptNIH-PA Author Manuscriptto be a formulaic set of rules, because the appropriate assays depend in part on the question beingasked and the system being used. In addition, we emphasize that no individual assay is guaranteedto be the most appropriate one in every situation, and we strongly recommend the use of multipleassays to verify an autophagic response.Keywords autolysosome; autophagosome; flux; lysosome; phagophore; stress; vacuole At the first Keystone Symposium on Autophagy in Health and Disease, one of the researchers in the audience, after listening to several comments detailing inadequacies in documenting autophagy, asked the question “What are the essential criteria for demonstrating autophagy?”This is a reasonable question, particularly considering that each of us may have his/her own opinion regarding the answer. Unfortunately, this presents something of a “moving target” for researchers who may think they have met those criteria, only to find out that the reviewer of their paper has different ideas. Conversely, as a reviewer, it is tiresome to raise the same objections repeatedly, wondering why researchers have not fulfilled some of the basic requirements for establishing the occurrence of an autophagic process. In addition, drugs that potentially modulate autophagy are increasingly being used in clinical trials, and screens are being carried out for new drugs that can modulate autophagy for therapeutic purposes. Clearly it is important to determine whether these drugs are truly affecting autophagy based on a set of accepted criteria. Accordingly, we describe here a basic set of contemporary guidelines that can be used by researchers to plan and interpret their experiments, by clinicians to decide which avenue of treatment is appropriate, and by both authors and reviewers to justify or criticize an experimental approach.Several fundamental points must be kept in mind as we establish guidelines for the selectionof appropriate methods to monitor autophagy. Importantly, there are no absolute criteria fordetermining the autophagic status that apply to every situation. This is because some assaysare inappropriate, problematic or may not work at all in particular cells, tissues or organisms.2 In addition, these guidelines may evolve as new methodologies are developed and currentassays of the process are superseded. Nonetheless, it is useful to establish guidelines foracceptable assays that can reliably monitor autophagy in many experimental systems. It isimportant to note that in this set of guidelines the term “autophagy” generally refers tomacroautophagy; other autophagy-related processes are specifically designated whenappropriate.An important point is that autophagy is a dynamic, multi-step process that can be modulatedat several steps, both positively and negatively. In this respect, the autophagic pathway is notdifferent from other cellular pathways. An accumulation of autophagosomes (be they measuredby electron microscopy (EM) image analysis, as fluorescent GFP-LC3 dots, or as LC3lipidation on a western blot), could, for example, reflect either increased autophagosomeformation due to increases in autophagic activity, or to reduced turnover of autophagosomes(Fig. 1). The latter can occur by inhibiting their maturation to amphisomes or autolysosomes,which happens if there are defects in fusion with endosomes or lysosomes, respectively, orfollowing inefficient degradation of the cargo once fusion has occurred.4 For the purposes ofthis review, the autophagic compartments are referred to as the sequestering(preautophagosomal) phagophore,5 the autophagosome,6 the amphisome (generated by fusionof autophagosomes with endosomes, also referred to as an acidic late autophagosome 7)8 andthe autolysosome (generated by fusion of autophagosomes or amphisomes with a lysosome,also referred to as an autophagolysosome).6 We note that the use of the term “phagophore” inthis review has no implied meaning in regard to the origin of the autophagosomal membrane.NIH-PA Author ManuscriptNIH-PA Author ManuscriptNIH-PA Author Manuscript。
德谷门冬双胰岛素注射液治疗2_型糖尿病的疗效及安全性研究
DOI：10.16658/ki.1672-4062.2023.19.084德谷门冬双胰岛素注射液治疗2型糖尿病的疗效及安全性研究戴卉，张开凤，朱凤丽江苏省镇江市丹徒区人民医院内分泌科，江苏镇江212000[摘要]目的探讨德谷门冬双胰岛素注射液在2型糖尿病中的效果以及安全性。
方法选取2022年1月—2023年7月江苏省镇江市丹徒区人民医院收治的62例2型糖尿病患者为研究对象，按随机数表法分为对照组（n=31）和观察组（n=31）。
对照组患者接受门冬胰岛素30注射液治疗，观察组患者接受德谷门冬双胰岛素注射治疗。
对比两组患者临床疗效、血糖变化和不良反应发生率。
结果观察组治疗有效为96.77%，高于对照组的77.42%，差异有统计学意义（χ2=5.167，P=0.023）。
治疗前，两组患者血糖水平比较，差异无统计学意义（P>0.05）；治疗后，两组患者血糖水平均改善，且观察组血糖指标低于对照组，差异有统计学意义（P< 0.05）。
观察组不良反应发生率低与对照组，差异有统计学意义（P<0.05）。
结论德谷门冬双胰岛素的应用可以明显改善2型糖尿病患者血糖水平，疗效更为确切，且安全性更高，不会增加用药后不良反应。
[关键词] 2型糖尿病；德谷门冬双胰岛素；门冬胰岛素30注射液；安全性[中图分类号] R587 [文献标识码] A [文章编号] 1672-4062（2023）10（a）-0084-04Study on the Efficacy and Safety of Insulin Degludec and Insulin Aspart Injection in the Treatment of Type 2 Diabetes MellitusDAI Hui, ZHANG Kaifeng, ZHU FengliDepartment of Endocrinology, Zhenjiang Dantu District People's Hospital, Zhenjiang, Jiangsu Province, 212000 China [Abstract] Objective To explore the effect and safety of insulin degludec and insulin aspart injection in type 2 diabe⁃tes mellitus.Methods 62 patients of type 2 diabetes mellitus patients admitted to Zhenjiang Dantu District People's Hospital, Jiangsu Province from January 2022 to July 2023 were selected as study objects and divided into the control group (n=31) and the observation group (n=31) by taking the random number table method. The patients in the control group were treated with insulin aspart 30 injection and the patients in the observation group were treated with insulin degludec and insulin aspart injection. Compared the clinical efficacy, the changes in blood glucose and the incidence of adverse reactions between the two groups of patients.Results The treatment effectiveness of the observation group was 96.77%, which was higher than that of the control group, which was 77.42%, and the difference was statistically significant (χ2=5.167, P=0.023). There was no statistically significant difference in blood glucose levels between the two groups before treatment (P>0.05). After treatment, blood glucose levels improved in both groups, and the level of blood glucose in the observation group were lower than those in the control group, and the difference was statistically significant (P<0.05). The incidence of adverse reactions in the observation group was lower than that in the control group, and the difference was statistically significant (P<0.05).Conclusion The application of insulin degludec and in⁃sulin aspart can significantly improve the blood glucose level of patients with type 2 diabetes mellitus, the efficacy is more accurate, and the safety is higher, and it will not increase the occurrence of adverse reactions after the use of medication.[作者简介]戴卉（1985-），女，本科，主治医师，研究方向为内分泌科。
219525882_超高效液相色谱法检测特殊医学用途配方食品中22种游离氨基酸
尹丽丽，郑红，程志，等. 超高效液相色谱法检测特殊医学用途配方食品中22种游离氨基酸[J]. 食品工业科技，2023，44（14）：264−271. doi: 10.13386/j.issn1002-0306.2022080065YIN Lili, ZHENG Hong, CHENG Zhi, et al. Determination of 22 Free Amino Acids in Food for Special Medical Purposes by Ultra-performance Liquid Chromatography[J]. Science and Technology of Food Industry, 2023, 44(14): 264−271. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2022080065· 分析检测 ·超高效液相色谱法检测特殊医学用途配方食品中22种游离氨基酸尹丽丽，郑 红，程 志，卢兰香，薛 霞，王 骏，胡 梅，刘艳明*（山东省食品药品检验研究院，山东省特殊医学用途配方食品质量控制工程技术研究中心，山东省食品药品安全检测工程技术研究中心，山东济南 250101）摘 要：本文建立了特殊医学用途配方食品中22种游离氨基酸的超高效液相色谱检测方法。
样品中氨基酸用磺基水杨酸提取，以2,4二硝基氟苯（DNFB ）为柱前衍生试剂，以乙腈水-甲酸铵为流动相梯度洗脱，HSS T3色谱柱高效分离，超高效液相-二极管阵列检测器检测，外标法定量。
结果显示，22种氨基酸在2~100 μg/mL 范围内线性关系良好（R 2>0.99）。
方法检出限（S/N ≥3）为10 mg/100 g ，定量限（S/N ≥10）为25 mg/100 g ，低、中、高三个加标水平的回收率在90.1%~109.1%，相对标准偏差为0.8%~6.9%。