PNU-282987_HNMR_18553_MedChemExpress
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.。
大动脉炎患者外周血单个核细胞RT-qPCR内参基因的选择
January 2021Vol.41 No.12021 年 1 月 第 41 卷 第 1 期基础医学与临床Basic & Clinical Medicine文章编号：1001-6325 ( 2021 ) 01-0087-06研究论文大动脉炎患者外周血单个核细胞RT-qPCR 内参基因的选择田苡箫，李菁*收稿日期：2019-11-18 修回日期：2020-04-30*通信作者(corresponding author ) ：lijing6515@ （中国医学科学院北京协和医学院北京协和医院风湿免疫科风湿免疫病学教育部重点实验室国家皮肤与免疫疾病临床医学研究中心，北京100032）扌摘要：目的筛选适于在大动脉炎（TAK ）患者和健康人群（HC ）之间比较外周血单个核细胞（PBMC ）中mRNA 表达水平的内参基因。
方法提取PBMC 中的总RNA,应用RT-qPCR ，分别采用geNorm 、NormFinder 、BestKeeper 3种软 件程序，分析 3-glucuronidase ,GAPDH ,ACTB ,SDHA ,HPRT1,RPL13A ,B2M , YWHAZ 和 PKG1 9 个基因的 mRNA 表达稳定性。
以T-bet 、GATA3和RORC 作为目的基因，比较不同稳定性的内参基因对mRNA 相对丰度的影响。
结果geNorm 筛选得到的基因组合为B 2M-SDHA , Nor^nFinder 和BestKeeper 筛选出最稳定的内参基因均为HPRT1 ； 3种方法均显示GAPDH 的稳定性较差。
结论自身免疫病患者在接受免疫抑制药物治疗时,原本稳定表达的基因可能会 上调或下调;在样本量较小时，稳定性更好的内参基因可能更有助于检测组间差异。
关键词：大动脉炎;实时定量聚合酶链式反应;RNA 稳定性；内参基因选择中图分类号:R593.2 文献标志码:AValidation of reference genes for the normalization of the RT-qPCRin peripheral blood mononuclear cells of patients with Takayasu arteritisTIAN Yi-xiao ， LI Jing *(Department of Rheumatology and Immunology , Key Laboratory of Rheumatology and Clinical Immunology , Ministry of Education ,National Clinical Research Center for Dermatologic and Immunologic Diseases ( NCRC-DID ),Peking Union Medical College Hospital , CAMS & PUMC , Beijing 100032, China)Abstract ： Objective To validate proper reference genes for quantitative real-time polymerase chain reaction ( RT-qPCR) used for comparing mRNA expression levels in Takayasu arteritis" (TAK) and healthy controls' ( HC ) pe ripheral blood mononuclear cells ( PBMC ). Methods Total RNA in PBMCs was extracted and used RT-qPCR to determine the profiles of 9 candidate genes , including 0-glucuronidase, GAPDH , ACTB , SDHA , HPRT1, RPL13A , B2M , YWHAZ and PKG1. Then compared their transcription stability by geNorm , NormFinder , and Best Keeper. Afterwards , with T-bet , GATA3 and RORC as the targeted genes , explored the influence of reference genes with different stability on mRNA relative abundance. Results The gene combination of B2M-SDHA was selected bygeNorm , and HPRT1 was the most stable one in analysis results of NormFinder and BestKeeper , while GAPDH was less stable. Conclusions Genes that have been expressed stably may be upregulated or downregulated whenpatients with autoimmune diseases received immunosuppressive drugs. When the sample size is small ， the more sta ble internal reference may facilitate the identification of inter-groups difference.Key words : Takayasu arteritis ； real-time polymerase chain reaction ； RNA stability ； selection of reference gene88基础医学与临床Basic&Clinical Medicine2021.41(1)反转录实时荧光定量聚合酶链式反应（reverse quantitative real-time polymerase chain reaction,RT-qPCR）是目前分析基因表达水平的黄金标准，却经常表现出重复性欠佳的问题，选取合适的内参基因有助于改善这一情况［1］。
血清C_肽与糖化血红蛋白联合检验在糖尿病临床诊断中的效果分析
· 医学检验 ·糖尿病新世界 2023年4月糖尿病新世界 DIABETES NEW WORLD血清C 肽与糖化血红蛋白联合检验在糖尿病临床诊断中的效果分析郑梅淋，田萍萍，钟秋蓉厦门市海沧医院检验科，福建厦门 361000[摘要] 目的 分析糖尿病临床诊断中应用血清C 肽联合糖化血红蛋白检验的效果。
方法 选取厦门市海沧医院2020年1月—2021年12月收治的200例疑似糖尿病患者为研究对象，将口服葡萄糖耐量试验作为诊断金标准，分别对患者施以血清C 肽检验和糖化血红蛋白检验，比较两组的血清C 肽、血清C 肽联合糖化血红蛋白检验结果。
结果 在200例疑似糖尿病患者中，糖耐量试验检查确诊患者180例，占比90.00%，血清C 肽检测下，阳性检出率为98.33%（177例），血清C 肽联合糖化血红蛋白检测下，阳性检出率为86.11%（155例）。
联合检测的灵敏度、特异度、准确率优于单一检测，差异有统计学意义（P <0.05）。
结论 血清C 肽联合糖化血红蛋白检验能够提升糖尿病临床诊断准确性，可为临床诊断提供更为科学的参考依据。
[关键词] 血清C 肽；糖化血红蛋白；糖尿病；诊断；准确度；联合检验[中图分类号] R59 [文献标识码] A [文章编号] 1672-4062（2023）04（b ）-0066-04Effect Analysis of Combined Test of Serum C-peptide and GlycosylatedHemoglobin in Clinical Diagnosis of Diabetes MellitusZHENG Meilin, TIAN Pingping, ZHONG QiurongDepartment of Laboratory Medicine, Xiamen Haicang Hospital, Xiamen, Fujian Province, 361000 China[Abstract ] Objective To analyze the effect of serum C-peptide combined with glycosylated hemoglobin test in the clinical diagnosis of diabetes. Methods We selected 200 patients with suspected diabetes who were admitted to Xia‐men Haicang Hospital from January 2020 to December 2021 as the research object, and took the oral glucose toler‐ance test as the diagnostic gold standard. Serum C-peptide test and Glycated hemoglobin test were performed on thepatients, and the results of serum C-peptide, serum C-peptide and Glycated hemoglobin test were compared betweenthe two groups. Results Among 200 suspected diabetes patients, 180 patients were diagnosed by Glucose tolerancetest, accounting for 90.00%. The positive detection rate was 98.33% (177 cases) by serum C-peptide test, and 86.11% (155 cases) by serum C-peptide combined with Glycated hemoglobin test. The sensitivity, specificity, and accuracy of combined detection were better than those of single detection, and the difference was statistically significant (P <0.05).Conclusion Serum C-peptide combined with Glycated hemoglobin test can improve the accuracy of clinical diagnosis of diabetes and provide more scientific reference for clinical diagnosis.[Key words ] Serum C-peptide; Glycosylated hemoglobin; Diabetes; Diagnosis; Accuracy; Combined test糖尿病是常见的慢性代谢性疾病，主要因为胰岛素代谢紊乱而引起的代谢性疾病，按照疾病成因，可分成1型糖尿病和2型糖尿病两种类型。
TMA-DPH-SDS-MedChemExpress
Inhibitors, Agonists, Screening LibrariesSafety Data Sheet Revision Date:Mar.-25-2019Print Date:Mar.-25-20191. PRODUCT AND COMPANY IDENTIFICATION1.1 Product identifierProduct name :TMA-DPHCatalog No. :HY-D0986CAS No. :115534-33-31.2 Relevant identified uses of the substance or mixture and uses advised againstIdentified uses :Laboratory chemicals, manufacture of substances.1.3 Details of the supplier of the safety data sheetCompany:MedChemExpress USATel:609-228-6898Fax:609-228-5909E-mail:sales@1.4 Emergency telephone numberEmergency Phone #:609-228-68982. HAZARDS IDENTIFICATION2.1 Classification of the substance or mixtureNot a hazardous substance or mixture.2.2 GHS Label elements, including precautionary statementsNot a hazardous substance or mixture.2.3 Other hazardsNone.3. COMPOSITION/INFORMATION ON INGREDIENTS3.1 SubstancesSynonyms:NoneFormula:C28H31NO3SMolecular Weight:461.62CAS No. :115534-33-34. FIRST AID MEASURES4.1 Description of first aid measuresEye contactRemove any contact lenses, locate eye-wash station, and flush eyes immediately with large amounts of water. Separate eyelids with fingers to ensure adequate flushing. Promptly call a physician.Skin contactRinse skin thoroughly with large amounts of water. Remove contaminated clothing and shoes and call a physician.InhalationImmediately relocate self or casualty to fresh air. If breathing is difficult, give cardiopulmonary resuscitation (CPR). Avoid mouth-to-mouth resuscitation.IngestionWash out mouth with water; Do NOT induce vomiting; call a physician.4.2 Most important symptoms and effects, both acute and delayedThe most important known symptoms and effects are described in the labelling (see section 2.2).4.3 Indication of any immediate medical attention and special treatment neededTreat symptomatically.5. FIRE FIGHTING MEASURES5.1 Extinguishing mediaSuitable extinguishing mediaUse water spray, dry chemical, foam, and carbon dioxide fire extinguisher.5.2 Special hazards arising from the substance or mixtureDuring combustion, may emit irritant fumes.5.3 Advice for firefightersWear self-contained breathing apparatus and protective clothing.6. ACCIDENTAL RELEASE MEASURES6.1 Personal precautions, protective equipment and emergency proceduresUse full personal protective equipment. Avoid breathing vapors, mist, dust or gas. Ensure adequate ventilation. Evacuate personnel to safe areas.Refer to protective measures listed in sections 8.6.2 Environmental precautionsTry to prevent further leakage or spillage. Keep the product away from drains or water courses.6.3 Methods and materials for containment and cleaning upAbsorb solutions with finely-powdered liquid-binding material (diatomite, universal binders); Decontaminate surfaces and equipment by scrubbing with alcohol; Dispose of contaminated material according to Section 13.7. HANDLING AND STORAGE7.1 Precautions for safe handlingAvoid inhalation, contact with eyes and skin. Avoid dust and aerosol formation. Use only in areas with appropriate exhaust ventilation.7.2 Conditions for safe storage, including any incompatibilitiesKeep container tightly sealed in cool, well-ventilated area. Keep away from direct sunlight and sources of ignition.Recommended storage temperature: -20°C, protect from lightShipping at room temperature if less than 2 weeks.7.3 Specific end use(s)No data available.8. EXPOSURE CONTROLS/PERSONAL PROTECTION8.1 Control parametersComponents with workplace control parametersThis product contains no substances with occupational exposure limit values.8.2 Exposure controlsEngineering controlsEnsure adequate ventilation. Provide accessible safety shower and eye wash station.Personal protective equipmentEye protection Safety goggles with side-shields.Hand protection Protective gloves.Skin and body protection Impervious clothing.Respiratory protection Suitable respirator.Environmental exposure controls Keep the product away from drains, water courses or the soil. Cleanspillages in a safe way as soon as possible.9. PHYSICAL AND CHEMICAL PROPERTIES9.1 Information on basic physical and chemical propertiesAppearance Light yellow to yellow (Solid)Odor No data availableOdor threshold No data availablepH No data availableMelting/freezing point No data availableBoiling point/range No data availableFlash point No data availableEvaporation rate No data availableFlammability (solid, gas)No data availableUpper/lower flammability or explosive limits No data availableVapor pressure No data availableVapor density No data availableRelative density No data availableWater Solubility No data availablePartition coefficient No data availableAuto-ignition temperature No data availableDecomposition temperature No data availableViscosity No data availableExplosive properties No data availableOxidizing properties No data available9.2 Other safety informationNo data available.10. STABILITY AND REACTIVITY10.1 ReactivityNo data available.10.2 Chemical stabilityStable under recommended storage conditions.10.3 Possibility of hazardous reactionsNo data available.10.4 Conditions to avoidNo data available.10.5 Incompatible materialsStrong acids/alkalis, strong oxidising/reducing agents.10.6 Hazardous decomposition productsUnder fire conditions, may decompose and emit toxic fumes.Other decomposition products - no data available.11.TOXICOLOGICAL INFORMATION11.1 Information on toxicological effectsAcute toxicityClassified based on available data. For more details, see section 2Skin corrosion/irritationClassified based on available data. For more details, see section 2Serious eye damage/irritationClassified based on available data. For more details, see section 2Respiratory or skin sensitizationClassified based on available data. For more details, see section 2Germ cell mutagenicityClassified based on available data. For more details, see section 2CarcinogenicityIARC: No component of this product present at a level equal to or greater than 0.1% is identified as probable, possible or confirmed human carcinogen by IARC.ACGIH: No component of this product present at a level equal to or greater than 0.1% is identified as a potential or confirmed carcinogen by ACGIH.NTP: No component of this product present at a level equal to or greater than 0.1% is identified as a anticipated or confirmed carcinogen by NTP.OSHA: No component of this product present at a level equal to or greater than 0.1% is identified as a potential or confirmed carcinogen by OSHA.Reproductive toxicityClassified based on available data. For more details, see section 2Specific target organ toxicity - single exposureClassified based on available data. For more details, see section 2Specific target organ toxicity - repeated exposureClassified based on available data. For more details, see section 2Aspiration hazardClassified based on available data. For more details, see section 212. ECOLOGICAL INFORMATION12.1 ToxicityNo data available.12.2 Persistence and degradabilityNo data available.12.3 Bioaccumlative potentialNo data available.12.4 Mobility in soilNo data available.12.5 Results of PBT and vPvB assessmentPBT/vPvB assessment unavailable as chemical safety assessment not required or not conducted.12.6 Other adverse effectsNo data available.13. DISPOSAL CONSIDERATIONS13.1 Waste treatment methodsProductDispose substance in accordance with prevailing country, federal, state and local regulations.Contaminated packagingConduct recycling or disposal in accordance with prevailing country, federal, state and local regulations.14. TRANSPORT INFORMATIONDOT (US)This substance is considered to be non-hazardous for transport.IMDGThis substance is considered to be non-hazardous for transport.IATAThis substance is considered to be non-hazardous for transport.15. REGULATORY INFORMATIONSARA 302 Components:No chemicals in this material are subject to the reporting requirements of SARA Title III, Section 302.SARA 313 Components:This material does not contain any chemical components with known CAS numbers that exceed the threshold (De Minimis) reporting levels established by SARA Title III, Section 313.SARA 311/312 Hazards:No SARA Hazards.Massachusetts Right To Know Components:No components are subject to the Massachusetts Right to Know Act.Pennsylvania Right To Know Components:No components are subject to the Pennsylvania Right to Know Act.New Jersey Right To Know Components:No components are subject to the New Jersey Right to Know Act.California Prop. 65 Components:This product does not contain any chemicals known to State of California to cause cancer, birth defects, or anyother reproductive harm.16. OTHER INFORMATIONCopyright 2019 MedChemExpress. The above information is correct to the best of our present knowledge but does not purport to be all inclusive and should be used only as a guide. The product is for research use only and for experienced personnel. It must only be handled by suitably qualified experienced scientists in appropriately equipped and authorized facilities. The burden of safe use of this material rests entirely with the user. MedChemExpress disclaims all liability for any damage resulting from handling or from contact with this product.Caution: Product has not been fully validated for medical applications. For research use only.Tel: 609-228-6898 Fax: 609-228-5909 E-mail: tech@Address: 1 Deer Park Dr, Suite Q, Monmouth Junction, NJ 08852, USA。
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］。
对比分析咪达唑仑与瑞马唑仑对全身麻醉下行腹腔镜胆囊切除术患者的效果及对术后认知功能的影响
DOI：10.19368/ki.2096-1782.2023.14.097对比分析咪达唑仑与瑞马唑仑对全身麻醉下行腹腔镜胆囊切除术患者的效果及对术后认知功能的影响陈虹烨，张鑫磊，佟飞，孔明健徐州矿务集团总医院麻醉科，江苏徐州221006[摘要]目的对比分析咪达唑仑与瑞马唑仑对全身麻醉下行腹腔镜胆囊切除术患者的效果及对术后认知功能的影响。
方法选取2022年3月—2023年5月徐州矿务集团总医院收治的全身麻醉下行腹腔镜胆囊切除术的患者92例为研究对象，根据随机数表法分为两组，各46例。
对照组麻醉诱导时静脉注射0.05 mg/kg咪达唑仑、0.50 µg/kg舒芬太尼、0.60 mg/kg罗库溴铵；观察组麻醉诱导时静脉注射0.30 mg/kg瑞马唑仑、0.50 µg/kg舒芬太尼、0.60 mg/kg罗库溴铵。
对比两组患者术后睁眼时间、拔管时间、麻醉复苏室停留时间，手术前后应激指标、认知功能及术后不良反应。
结果观察组术后睁眼、拔管及麻醉复苏室停留时间均短于对照组，差异有统计学意义（P<0.05）。
术后，观察组胰岛素、皮质醇水平均低于对照组，差异有统计学意义（P<0.05）。
术后24、72 h，观察组简易智力状态检查量表评分均高于对照组，差异有统计学意义（P<0.05）。
观察组患者呼吸抑制、低血压、嗜睡、恶心呕吐等的不良反应发生率为10.87%，低于对照组的28.26%，差异有统计学意义（χ2= 4.420，P<0.05）。
结论采取瑞马唑仑对全身麻醉下行腹腔镜胆囊切除术患者进行麻醉可以加速患者术后麻醉恢复，麻醉效果较好，还可降低机体应激反应，较快恢复术后认知功能，且安全性较高。
[关键词]咪达唑仑；瑞马唑仑；全身麻醉；腹腔镜胆囊切除术；应激反应；术后认知功能[中图分类号]R657 [文献标识码]A [文章编号]2096-1782（2023）07(b)-0097-04Comparative Analysis of the Effects of Midazolam and Remimazolam on Patients Undergoing Laparoscopic Cholecystectomy under General Anes⁃thesia and Their Impact on Postoperative Cognitive FunctionCHEN Hongye, ZHANG Xinlei, TONG Fei, KONG MingjianDepartment of Anesthesiology, Xuzhou Mining Group General Hospital, Xuzhou, Jiangsu Province, 221006 China[Abstract] Objective To comparatively analyze the effects of midazolam and remimazolam on patients undergoing laparoscopic cholecystectomy under general anesthesia and their impact on postoperative cognitive function. Methods 92 patients undergoing laparoscopic cholecystectomy under general anesthesia admitted to Xuzhou Mining Group Gen‐eral Hospital from March 2022 to May 2023 were selected as the study subjects. They were divided into two groups us‐ing a random number table method, with 46 patients in each group. Intravenous injection of 0.05 mg/kg midazolam and 0.50 µg/kg sufentanil and 0.60 mg/kg rocuronium during anesthesia induction in the control group; Observation group received intravenous injection of 0.30 mg/kg remimazolam and 0.50 µg/kg sufentanil and 0.60 mg/kg ro‐curonium during anesthesia induction. Compared the postoperative eye opening time, extubation time, anesthesia re‐suscitation room stay time, preoperative and postoperative stress indicators, cognitive function, and postoperative ad‐verse reactions between two groups of patients. Results The observation group had shorter postoperative eye opening, extubation, and anesthesia resuscitation room stay times than the control group, the difference was statistically signifi‐cant (P<0.05). After surgery, the levels of INS and Cor in the observation group were lower than those in the control group, the difference was statistically significant (P<0.05). At 24 and 72 hours after surgery, the MMSE scores in the[作者简介] 陈虹烨（1985-），女，本科，副主任医师，研究方向为临床麻醉。
益生菌对阿尔茨海默病作用的研究进展
益生菌对阿尔茨海默病作用的研究进展发布时间：2021-12-14T06:08:15.523Z 来源：《中国结合医学杂志》2021年12期作者：宋鑫萍1,2，李盛钰2，金清1[导读] 阿尔茨海默病已成为威胁全球老年人生命健康的主要疾病之一，患者数量逐年攀升，其护理的经济成本高，给全球经济造成重大挑战。
近年来研究显示，益生菌在适量使用时作为有益于宿主健康的微生物，在防治阿尔茨海默病方面具有积极影响，其作用机制可能通过调节肠道菌群，影响神经免疫系统，调控神经活性物质以及代谢产物，通过肠-脑轴影响该病发生和发展。
宋鑫萍1,2，李盛钰2，金清11.延边大学农学院，吉林延吉 1330022.吉林省农业科学院农产品加工研究所，吉林长春 130033摘要：阿尔茨海默病已成为威胁全球老年人生命健康的主要疾病之一，患者数量逐年攀升，其护理的经济成本高，给全球经济造成重大挑战。
近年来研究显示，益生菌在适量使用时作为有益于宿主健康的微生物，在防治阿尔茨海默病方面具有积极影响，其作用机制可能通过调节肠道菌群，影响神经免疫系统，调控神经活性物质以及代谢产物，通过肠-脑轴影响该病发生和发展。
本文综述了近几年来国内外益生菌对阿尔茨海默病的作用进展，以及其预防和治疗阿尔茨海默病的潜在作用机制。
关键词：益生菌；阿尔茨海默病；肠道菌群；机制Recent Progress in Research on Probiotics Effect on Alzheimer’s DiseaseSONG Xinping1,2，LI Shengyu2，JI Qing1*(1.College of Agricultural, Yanbian University, Yanji 133002，China)(2.Institute of Agro-food Technology, Jilin Academy of Agricultural Sciences, Chanchun 130033, China)Abstract：Alzheimer’s disease has become one of the major diseases threatening the life and health of the global elderly. The number of patients is increasing year by year, and the economic cost of nursing is high, which poses a major challenge to the global economy. In recent years, studies have shown that probiotics, as microorganisms beneficial to the health of the host, have a positive impact on the prevention and treatment of Alzheimer’s disease. Its mechanism may be through regulating intestinal flora, affecting the nervous immune system, regulating the neuroactive substances and metabolites, and affecting the occurrence and development of the disease through thegut- brain axis. This paper reviews the progress of probiotics on Alzheimer’s disease at home and abroad in recent years, as well as its potential mechanism of prevention and treatment.Key words：probiotics; Alzheimer’s disease; gut microbiota; mechanism阿尔茨海默病（Alzheimer’s disease, AD），系中枢神经系统退行性疾病，属于老年期痴呆常见类型，临床特征主要包括：记忆力减退、认知功能障碍、行为改变、焦虑和抑郁等。
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。
基质分散固相萃取净化液相色谱检测谷物中赭曲霉毒素A
Ｃ１ ｓ ｌ ｈ ｓ ｎ ａ ｌｓ ｐ ｗｄ ｒ ｐ ａ ｅ ｎ ｔ ｅ ｍｏ ｔ ｒ ｇ ｎ ｌ ｎ ｕ ｌ ｘ ｄ ｂ ｅ ｔｅ Ａｎ ｈ ｎ ｐ ｔｉ ｌ ｓ ｙ ｉ ｇ ｓ ｗｉｈ ８ ｏ ｉ ｐ ａ ｅ ａ ｄ ｓ ｍｐ ｅ ｏ ｅ ｌ ｃ ｄ ｉ ｈ ｒａ ｅ ｔｙ ａ ｄ ｆ ｌ ｍｉ ｅ ｙ ｐ ｓｌ ． ｄ ｔ ｅ ｕ ｎ ｇ ａ ｓ ｓ ｒｎ ｅ ｔ ｄ ｙ
测赭 曲霉 毒素 Ａ 含量 的方 法 。
目前 报道 的检 测 赭 曲 霉 毒 素 Ａ 含 量 的 主要 方 法 有 ：薄 层 色 谱 法 （ Ｃ）７、酶 联 免 疫 法 ＴＩ ＿ Ｊ （ Ｉ Ｓ ］免 疫胶 体 金 试 纸 条 法ｌ 和 高 效 液 色 谱 Ｅ Ａ） 、 Ｉ ｇ 法 （ Ｉ ）ｌ。 。薄层 色谱 分析 法是 较早 用于毒 素 ＨＰ ｌ 等 ｃ 。 检 测 的一种方 法 ， 其优 点是方 法 简单 ， 用 的试剂 价 使
ＡＢＳ ＴＲＡＣＴ ： ｅ ｔ ｂｌ ｈ ｍ ｅ ｈｏ ｏｆ ｘｔａ ｔｏｎ Ｔｏ ｓ ａ ｉ ａ ｓ ｔ ｄ ｅ ｒ ｃ ｉ ｗ ｉｈ ｍａｔｉ ｓ ｉ— ｔ ｒｘ ｏｌｄ ｐｈａ ｅ ｓ ｄｉｐ ｒ ｉｎ （Ｍ Ｓ ｓ ｅ ｓｏ ＰＤ ），ｈｉ ｐｅ ｆ ｍ ａ ｅ ｌｑ ｄ ｇｈ ｒｏｒ ｎｃ ｉ ｕｉ
４ ０ｎ ； 样 量 ２ ｌ ６ ｍ 进 ０“ 。
差 ， 放 时 间短 ； 体 金 颗 粒 大 ， 透 能 力 较 差 等 。 存 胶 渗
食 用 小 麦 样 品 中 赭 曲 霉 索 Ａ 的 污 染 率 为 ４ ． ６／ ５ １ ６ ９。
收 稿 日期 ：Ｏ ２Ｏ ３ 修 回 日期 ：０ ２０ ２ ｌ一３２ ； ２ １ －７１ ９ 作者简介 ： 李 尧 （ ９ １） 男 ， 析 员 ， 程 师 ， １ ８一 ， 分 工 主要 从 事 色 谱 、 谱分 析 。 质
细胞程序性死亡配体1表位肽疫苗的设计及抗肿瘤活性
学 报Journal of China Pharmaceutical University 2023，54（2）：245 - 254245细胞程序性死亡配体1表位肽疫苗的设计及抗肿瘤活性邵世帅，段树康，田浤，姚文兵，高向东*（中国药科大学生命科学与技术学院江苏省生物药物成药性研究重点实验室，南京 211198）摘 要 目前已有多款细胞程序性死亡受体1（PD-1）和其配体（PD-L1）免疫检查点阻断抗体用于临床治疗，但只有部分患者表现出临床反应，因此需要一种替代的肿瘤免疫治疗策略。
以PD-L1为靶点的治疗性肿瘤疫苗是一种有意义的尝试。
本研究设计了以PD-L1为靶点的表位肽疫苗，然后基于人源化免疫系统（HIS）小鼠模型筛选，具有免疫原性的PD-L1表位肽，进一步研究其抗肿瘤活性。
结果显示，设计并筛选得到的PD-L1-B1表位肽疫苗不仅成功诱导了PD-L1特异性的体液免疫和细胞免疫，还表现出抗肿瘤活性。
此外，免疫治疗还增加了肿瘤的T淋巴细胞浸润，重塑了肿瘤免疫抑制微环境。
综上所述，PD-L1-B1表位肽疫苗表现出强效的抗肿瘤活性，对PD-1/PD-L1阻断不敏感的患者可能是一种有效的替代免疫治疗策略。
关键词肿瘤免疫；免疫检查点；PD-L1；表位肽疫苗中图分类号R73-3 文献标志码 A 文章编号1000 -5048（2023）02 -0245 -10doi：10.11665/j.issn.1000 -5048.2023022803引用本文邵世帅，段树康，田浤，等.细胞程序性死亡配体1表位肽疫苗的设计及抗肿瘤活性［J］.中国药科大学学报，2023，54（2）：245–254.Cite this article as：SHAO Shishuai，DUAN Shukang，TIAN Hong，et al. Design and antitumor activity of programmed cell death ligand 1 epitope peptide vaccine［J］.J China Pharm Univ，2023，54（2）：245–254.Design and antitumor activity of programmed cell death ligand 1 epitope peptide vaccineSHAO Shishuai, DUAN Shukang, TIAN Hong, YAO Wenbing, GAO Xiangdong*Jiangsu Provincial Key Laboratory of Druggability of Biopharmaceuticals, School of Life Science and Technology, China Pharmaceu⁃tical University, Nanjing 211198, ChinaAbstract Several programmed cell death protein 1 (PD-1) or its ligand (PD-L1) immune checkpoint blocking antibodies are available for clinical treatment, but only some patients show clinical response, so an alternative strategy for tumor immunotherapy is needed.A therapeutic tumor vaccine targeting PD-L1 is a meaningful attempt.In this study, we designed an epitope peptide vaccine targeting PD-L1, and then screened the immuno‑genic PD-L1 epitope peptide based on the humanized immune system (HIS) mouse model and further investigated its anti-tumor activity.The results show that the designed and screened PD-L1-B1 epitope peptide vaccine not only successfully induced PD-L1-specific humoral and cellular immunity, but also exhibit anti-tumor activity.In addi‑tion, immunotherapy increased T-lymphocyte infiltration of tumors and reshaped the tumor immunosuppressive microenvironment.In conclusion, PD-L1-B1 epitope peptide vaccine exhibits potent anti-tumor activity and may be an effective alternative immunotherapeutic strategy for patients insensitive to PD-1/PD-L1 blockade.Key words tumor immunity; immune checkpoint; PD-L1; epitope peptide vaccineThis study was supported by the National Natural Science Foundation of China (No.82073754, No.81973222); and the Key Research and Development Program of Xinjiang Uygur Autonomous Region (No.2020B03003)收稿日期2023-02-28 *通信作者Tel：134****2857E-mail：1019830864@基金项目国家自然科学基金资助项目（No.82073754，No.81973222）；新疆维吾尔自治区重点研发计划（No.2020B03003）学 报 Journal of China Pharmaceutical University 2023，54（2）：245 - 254第54 卷免疫检查点阻断（ICB ）疗法在癌症治疗方面取得了前所未有的进展，极大地改变了肿瘤治疗策略［1-2］。
超高效液相色谱-串联质谱法同时测定鸡肉中的6种磺胺类药物
超高效液相色谱-串联质谱法同时测定鸡肉中的6种磺胺类药物王红梅（山东省食用植物油质量检验中心山东省莒南县检验检测中心山东临沂276000）试验旨在建立同时快速检测鸡肉中6种磺胺类药物含量的超高效液相色谱串联质谱法。
鸡肉样品用乙腈超声提取，经Thermo Fisher Scientific Hyperis11Gold (100×2.1mm ，1.9μm)色谱柱分离，流动相A ：乙腈；流动相B ：0.1%甲酸水溶液作为流动相梯度洗脱。
电喷雾离子源正离子多反应监测模式定量分析。
结果表明，6种磺胺类药物在浓度10～500ng/mL 范围内线性关系很好，相关系数R 2≥0.9998。
6种磺胺类药物在含量为8和60μg/kg 的样品中，回收率为73%～88%，相对标准偏差RSD 为0.9%～6%。
综上，该方法高效、快速、高分辨，可以用于鸡肉样品中磺胺类药的定性、定量分析。
肉；磺胺类药物；超高效液相色谱-串联质谱磺胺类药物(sulfonamides ，SAs)是指一类具有对氨基苯磺酰胺结构化学药物的总称，用于预防和治疗全身性细菌感染性疾病，是当前畜禽生产中常用的抗菌、抗原虫药物，具有抗菌谱广、价格低、化学性质稳定、高效、低毒、使用方便等优点。
其低剂量使用时，可提高饲料的利用率，促进畜禽的生长，但不正确的用药量、用药时间、方式以及用药部位会导致磺胺在畜禽体内及其排泄物的过量残留，导致磺胺类在人体中蓄积，因此产生过敏反应、泌尿系统损害、抑制白细胞生成等危害，重者导致急性血管性水肿、休克甚至死亡[1]。
因此，我国农业部第235号公告《动物性食品中兽药最高残留限量》对SAs 总量及单个SAs 残留量做了明确规定，均不得超过100μg/kg 。
目前，测定动物源性食品中磺胺类药物的方法不止一种，但超高效液相色谱－串联质谱法背景干扰少，能高效、快速、高分辨的定性、定量检测和自动分析微量物质，为高灵敏测定磺胺类药物提供了支持。
致病疫霉菌效应蛋白Pi05440毒性功能验证及寄主候选靶标筛选
核农学报2024，38（3）：0434~0442Journal of Nuclear Agricultural Sciences致病疫霉菌效应蛋白Pi05440毒性功能验证及寄主候选靶标筛选张蝶陈胜男赵迪王荟洁王洪洋 *刘晶 *（云南师范大学，云南省马铃薯生物学重点实验室，云南昆明650500）摘要：致病疫霉菌（Phytophthora infestans）侵染时会分泌大量效应蛋白进入寄主细胞，通过操纵寄主靶标抑制植物免疫反应。
为研究致病疫霉菌效应蛋白Pi05440的功能，本研究重点分析了Pi05440的蛋白特征、毒性功能及鉴定其寄主靶标；利用生物信息学数据库，预测Pi05440的保守结构域和信号肽。
构建pRI101-GFP-Pi05440表达载体用于Pi05440的亚细胞定位和毒性功能分析；同时，利用酵母双杂交技术对Pi05440的寄主靶标蛋白进行了筛选和鉴定。
结果表明，Pi05440基因全长969 bp，编码322个氨基酸。
结构预测结果显示Pi05440含有3个典型的KAZAL功能域。
亚细胞定位结果表明Pi05440定位在质膜和细胞间隙。
在本氏烟中瞬时表达该基因显著促进致病疫霉菌扩展。
通过酵母双杂交筛选，初步鉴定到3个Pi05440的候选靶标蛋白，分别为马铃薯过氧化氢酶12、马铃薯几丁质酶以及马铃薯MYB-like A蛋白。
本研究为探究致病疫霉菌效应蛋白Pi05440及其靶标蛋白如何调控植物免疫提供了重要线索。
关键词：致病疫霉菌；效应蛋白；亚细胞定位；酵母双杂交；靶标蛋白DOI：10.11869/j.issn.1000‑8551.2024.03.0434马铃薯（Solanum tuberosum）富含碳水化合物、维生素、矿物质和抗氧化物质等营养成分，对人类健康和可持续农业发展至关重要［1］。
然而，马铃薯生产过程中会面临许多病害的威胁，其中最为严重的是由致病疫霉菌（Phytophthora infestans）引起的晚疫病。
15523666_迷迭香酸对哮喘小鼠氧化性肺损伤的保护作用
$ 材 料 与 方 法
$C$ 材 料 !3!3!试验动物!0""8XDC[ 级雌性 &;B&&S 小鼠购自 广 西 医 科 大 学 实 验 动 物 中 心"动 物 合 格 证
号!D#2*%桂'"8!$?888"# 试 验 前 适 应 性 饲 养:P" 自由饮水 饮 食"饲 养 环 境 温 度 "$ c l! c"相 对 湿 度 $85 "085 # !3!3" 主 要 试 剂 和 药 品 9,D$D,'$1DZ?CQ 检测试 剂 盒 均 购 自 碧 云 天 生 物 技 术 研 究 所*,(; 购自 DEX\@ 公 司*液 态 铝 佐 剂 购 自 >OHM\I 公 司# 肿节风药材购自广 西 玉 林 中 桂 药 材 有 限 公 司"药 材
%!3广西大学动物科学技术学院"南宁 4:8884*"3广西畜牧研究所"南宁 4:888!'
摘要为评价迷迭香酸对哮 喘 小 鼠 模 型 氧 化 性 肺 损 伤 的 保 护 作 用"本 研 究 用 卵 清 蛋 白 %,(;'致 敏$激 发 雌 性 &;B&&S小鼠建立哮喘模型"并用 ,(; 和 Z"," 联合激发小鼠作为氧化肺损伤阳性对照模型#在最后一次滴鼻激 发 "$O 后 "取 支 气 管 肺 泡 灌 洗 液 %&;B['进 行 细 胞 计 数 并 测 定 活 性 氧 %9,D'$超 氧 化 物 歧 化 酶 %D,''和 谷 胱 甘 肽 过 氧化物酶%1DZ?CQ'水平"取左侧肺脏固定做 Z= 染色#结果显示"迷迭香酸可明显减少 &;B[ 中 细 胞 总 数 和 嗜 酸 性粒细胞数目"显著抑制肺组织和 &;B[ 中 9,D的产生"升高 D,' 和 1DZ?CQ水 平"改 善 肺 组 织 病 理 变 化# 本 试 验 结 果 表 明 "迷 迭 香 酸 对 氧 化 肺 损 伤 起 到 明 显 的 保 护 作 用 # 关 键 词 迷 迭 香 酸 *哮 喘 *氧 化 性 肺 损 伤 *抗 氧 化 中 图 分 类 号 9"0434 文 献 标 识 码 ; 文 章 编 号 !+6!?6":+%"8!6'!"?:+48?8+
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。
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。
高效液相色谱法测定小麦粉中赭曲霉毒素A的不确定度评定
分析检测高效液相色谱法测定小麦粉中赭曲霉毒素A的不确定度评定唐 莉（镇江市食品药品监督检验中心，江苏镇江 212000）摘 要：按照《食品安全国家标准食品中赭曲霉毒素A的测定》（GB 5009.96—2016）检测小麦粉中赭曲霉毒素A的含量，并建立不确定度分析的数学模型，对检验结果的不确定度来源进行分析及量化评定。
分析结果显示，标准曲线拟合和标准溶液的配制引入的不确定度最大。
小麦粉中赭曲霉毒素A的含量为4.62 μg·kg-1时，其扩展不确定度为0.31 μg·kg-1，k=2。
关键词：赭曲霉毒素A；不确定度；小麦粉；高效液相色谱法Evaluation of Uncertainty in the Determination of OchratoxinA in Wheat Flour by HPLCTANG Li(Zhenjiang Food and Drug Supervision and Inspection Center, Zhenjiang 212000, China) Abstract: According to GB 5009.96—2016, the content of ochratoxin A in wheat flour was detected, and a mathematical model for uncertainty analysis was established to analyze and quantitatively evaluate the sources of uncertainty in the test results. The analysis results show that the fitting of standard curves and the preparation of standard solutions introduce the greatest uncertainty. The content of ochratoxin A in wheat flour is 4.62 μg·kg-1, its expanded uncertainty is 0.31 μg·kg-1, k=2.Keywords: ochratoxin A; uncertainty; wheat flour; high performance liquid chromatography赭曲霉毒素A（Ochratoxin A，OTA）是一种分布广，与人类健康密切相关的真菌毒素，主要表现为慢性毒性，且具有很强的肝脏和肾脏毒性，并有致畸、致癌性，已受到全世界的广泛关注。