Mivebresib_COA_21605_MedChemExpress
米托蒽醌抑制USP11促进BACE1降解延缓阿尔茨海默病发展的实验研究
米托蒽醌抑制USP11促进BACE1降解延缓阿尔茨海默病发展的实验研究吴昌安;曹岐新;艾宗耀【期刊名称】《健康研究》【年(卷),期】2022(42)3【摘要】目的探讨米托蒽醌(Mitoxantrone,MTX)抑制USP11促进BACE1降解,从而延缓阿尔茨海默病(Alzheimer’s disease,AD)发展的作用机制。
方法构建USP11-siRNA-H4细胞株,Westen blot检测USP11、β-分泌酶(BACE1)等的表达水平;免疫共沉淀和蛋白质泛素化实验探究USP11和BACE1蛋白之间的泛素化调控作用。
以米托蒽醌(MTX)腹腔注射AD小鼠,水迷宫实验观其空间记忆能力,利用Westen blot与ELISA分别检测小鼠脑组织中USP11、BACE1与Aβ_(1-40)表达水平。
结果USP11-siRNA-H4细胞中USP11、BACE1的表达水平分别为(0.300±0.114)、(0.267±0.125),均低于NT siRNA组,差异有统计学意义(P<0.01)。
USP11与BACE1之间存在相互作用且可对BACE1去泛素化。
MTX处理后,AD小鼠记忆能力提高,且下调USP11、BACE1及Aβ_(1-40)的表达。
结论米托蒽醌通过抑制USP11促进BACE1泛素化与降解,从而降低Aβ_(1-40)的表达,进而可对阿尔茨海默病的神经病变起到延缓作用。
【总页数】5页(P297-301)【作者】吴昌安;曹岐新;艾宗耀【作者单位】浙江中医药大学附属湖州中医院神经内科【正文语种】中文【中图分类】R363.1【相关文献】1.Amiloride 通过抑制缺氧诱导的 NHE1表达而延缓 calpain 介导的 ABCA1降解2.PKCβ2途径抑制药物LY333531延缓高糖诱导心肌纤维化的实验研究3.醌氧化还原酶1抑制剂双香豆素促进HBV X蛋白降解进而抑制cccDNA转录的机制研究4.脑部渗透性ACE抑制剂可延缓阿尔茨海默病(Alzheimer disease)5.抑制胶原纤维合成延缓失神经支配骨骼肌萎缩的实验研究因版权原因，仅展示原文概要，查看原文内容请购买。
碧云天生物技术 Beyotime Biotechnology 生物素标记EMSA探针说明书
碧云天生物技术/Beyotime Biotechnology订货热线：400-168-3301或800-8283301订货e-mail：******************技术咨询：*****************网址：碧云天网站微信公众号生物素标记EMSA探针－β-Catenin/TCF (0.2μM)产品编号产品名称包装GS018B 生物素标记EMSA探针－β-Catenin/TCF (0.2µM) 200µl产品简介：生物素标记EMSA探针－β-Catenin/TCF是用于EMSA(也称gel shift)研究的生物素(Biotin)标记的β-Catenin/TCF consensus oligonucleotide。
这个生物素标记的双链寡核苷酸含有公认的β-Catenin/TCF结合位点，可以用作EMSA研究时的探针。
β-Catenin/TCF consensus oligo的序列如下：5'-CCC TTT GAT CTT ACC-3'3'-GGG AAA CTA GAA TGG-5'本生物素标记EMSA探针已经过纯化，可以直接用于EMSA结合反应。
本生物素标记EMSA探针可以和碧云天的化学发光法EMSA试剂盒(GS009)配套使用。
一个包装的生物素标记探针可以进行约200-400个样品的EMSA检测。
包装清单：产品编号产品名称包装GS018B 生物素标记EMSA探针－β-Catenin/TCF (0.2µM) 200µl—说明书1份保存条件：-20ºC保存，一年有效。
注意事项：避免加热到40ºC以上，温度过高会导致双链DNA探针解聚成单链。
而单链无法用于EMSA研究。
对于基于生物素标记的EMSA检测的详细操作可以参考碧云天的化学发光法EMSA试剂盒(GS009)的使用说明。
本产品仅限于专业人员的科学研究用，不得用于临床诊断或治疗，不得用于食品或药品，不得存放于普通住宅内。
USP60
USP60USP60 (Ubiquitin Specific Peptidase 60) is a deubiquitinating enzyme that plays a crucial role in cellular processes such as protein degradation, DNA repair, and cell cycle regulation. In this document, we will explore the structure, function, and regulatory mechanisms of USP60.Structure of USP60USP60 is a member of the ubiquitin-specific protease family. It consists of several distinct domains that contribute to its enzymatic activity and substrate specificity. The catalytic domain of USP60 contains a conserved cysteine residue that mediates the cleavage of the isopeptide bond between ubiquitin and its target protein. Adjacent to the catalytic domain, there is a ZnF-UBP domain that helps in substrate recognition and binding. The C-terminal domain of USP60 is responsible for its stability and localization within the cell.Function of USP60USP60 primarily functions as a deubiquitinating enzyme, deconjugating ubiquitin molecules from target proteins. By removing ubiquitin chains, USP60 can prevent the degradation of target proteins by the proteasome, thereby stabilizing them. Additionally, USP60 regulates the levels of ubiquitin by recycling ubiquitin molecules back into the cytoplasm for further use.USP60 is also involved in DNA repair processes. It interacts with various DNA repair proteins, such as BRCA1 and RAD51,and helps in the removal of ubiquitin moieties from DNA repair complexes. This activity of USP60 ensures efficient DNA damage repair and maintenance of genomic stability.Furthermore, USP60 has been implicated in cell cycle regulation. It interacts with cell cycle regulators, such as p53 and CDC25C, and modulates their stability and activity. This regulation of cell cycle progression by USP60 is essential for proper cell division and prevention of aberrant cell growth.Regulation of USP60 ActivityThe activity of USP60 is regulated by various mechanisms, including post-translational modifications, protein-protein interactions, and subcellular localization. One of the key regulatory mechanisms is phosphorylation. Phosphorylation of specific residues within USP60 can either enhance or inhibit its enzymatic activity. For example, phosphorylation of USP60 by ATM kinase increases its activity and promotes DNA repair.USP60 can also interact with other proteins, forming complexes that modulate its activity. For instance, interaction with BRCA1 enhances the deubiquitinating activity of USP60 in DNA repair processes. On the other hand, interaction with negative regulators such as UAF1 can inhibit the function of USP60.Subcellular localization of USP60 is crucial for its proper function. It is primarily localized in the nucleus, where it interacts with DNA repair and cell cycle proteins. However, under certain cellular stress conditions, USP60 can translocate to the cytoplasm and participate in other cellular processes.Role of USP60 in DiseaseDysregulation of USP60 function has been associated with various diseases. For example, aberrant expression or activity of USP60 has been observed in certain types of cancer. In some cases, overexpression of USP60 can stabilize oncoproteins and promote tumor growth. In other cases, downregulation of USP60 can impair DNA repair mechanisms, leading to genomic instability and increased cancer susceptibility.Furthermore, mutations in the USP60 gene have been associated with neurodegenerative disorders. These mutations can lead to a loss of USP60 function, resulting in the accumulation of abnormal proteins and neuronal cell death.ConclusionUSP60 is a versatile deubiquitinating enzyme that has important roles in protein degradation, DNA repair, and cell cycle regulation. Its structure, function, and regulatory mechanisms make it an attractive target for therapeutic interventions in various diseases. Further research is required to fully elucidate the complex network of interactions and pathways involving USP60.。
稳定性英文版
HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationFLUOXETINE HClC17H18F3NO•HClM.W. = 345.79CAS — 59333-67-4STABILITY INDICATINGA S S A Y V A L I D A T I O NMethod is suitable for:ýIn-process controlþProduct ReleaseþStability indicating analysis (Suitability - US/EU Product) CAUTIONFLUOXETINE HYDROCHLORIDE IS A HAZARDOUS CHEMICAL AND SHOULD BE HANDLED ONLY UNDER CONDITIONS SUITABLE FOR HAZARDOUS WORK.IT IS HIGHLY PRESSURE SENSITIVE AND ADEQUATE PRECAUTIONS SHOULD BE TAKEN TO AVOID ANY MECHANICAL FORCE (SUCH AS GRINDING, CRUSHING, ETC.) ON THE POWDER.ED. N0: 04Effective Date:APPROVED::HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationTABLE OF CONTENTS INTRODUCTION........................................................................................................................ PRECISION............................................................................................................................... System Repeatability ................................................................................................................ Method Repeatability................................................................................................................. Intermediate Precision .............................................................................................................. LINEARITY................................................................................................................................ RANGE...................................................................................................................................... ACCURACY............................................................................................................................... Accuracy of Standard Injections................................................................................................ Accuracy of the Drug Product.................................................................................................... VALIDATION OF FLUOXETINE HCl AT LOW CONCENTRATION........................................... Linearity at Low Concentrations................................................................................................. Accuracy of Fluoxetine HCl at Low Concentration..................................................................... System Repeatability................................................................................................................. Quantitation Limit....................................................................................................................... Detection Limit........................................................................................................................... VALIDATION FOR META-FLUOXETINE HCl (POSSIBLE IMPURITIES).................................. Meta-Fluoxetine HCl linearity at 0.05% - 1.0%........................................................................... Detection Limit for Fluoxetine HCl.............................................................................................. Quantitation Limit for Meta Fluoxetine HCl................................................................................ Accuracy for Meta-Fluoxetine HCl ............................................................................................ Method Repeatability for Meta-Fluoxetine HCl........................................................................... Intermediate Precision for Meta-Fluoxetine HCl......................................................................... SPECIFICITY - STABILITY INDICATING EVALUATION OF THE METHOD............................. FORCED DEGRADATION OF FINISHED PRODUCT AND STANDARD..................................1. Unstressed analysis...............................................................................................................2. Acid Hydrolysis stressed analysis..........................................................................................3. Base hydrolysis stressed analysis.........................................................................................4. Oxidation stressed analysis...................................................................................................5. Sunlight stressed analysis.....................................................................................................6. Heat of solution stressed analysis.........................................................................................7. Heat of powder stressed analysis.......................................................................................... System Suitability stressed analysis.......................................................................................... Placebo...................................................................................................................................... STABILITY OF STANDARD AND SAMPLE SOLUTIONS......................................................... Standard Solution...................................................................................................................... Sample Solutions....................................................................................................................... ROBUSTNESS.......................................................................................................................... Extraction................................................................................................................................... Factorial Design......................................................................................................................... CONCLUSION...........................................................................................................................ED. N0: 04Effective Date:APPROVED::HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationBACKGROUNDTherapeutically, Fluoxetine hydrochloride is a classified as a selective serotonin-reuptake inhibitor. Effectively used for the treatment of various depressions. Fluoxetine hydrochloride has been shown to have comparable efficacy to tricyclic antidepressants but with fewer anticholinergic side effects. The patent expiry becomes effective in 2001 (US). INTRODUCTIONFluoxetine capsules were prepared in two dosage strengths: 10mg and 20mg dosage strengths with the same capsule weight. The formulas are essentially similar and geometrically equivalent with the same ingredients and proportions. Minor changes in non-active proportions account for the change in active ingredient amounts from the 10 and 20 mg strength.The following validation, for the method SI-IAG-206-02 , includes assay and determination of Meta-Fluoxetine by HPLC, is based on the analytical method validation SI-IAG-209-06. Currently the method is the in-house method performed for Stability Studies. The Validation was performed on the 20mg dosage samples, IAG-21-001 and IAG-21-002.In the forced degradation studies, the two placebo samples were also used. PRECISIONSYSTEM REPEATABILITYFive replicate injections of the standard solution at the concentration of 0.4242mg/mL as described in method SI-IAG-206-02 were made and the relative standard deviation (RSD) of the peak areas was calculated.SAMPLE PEAK AREA#15390#25406#35405#45405#55406Average5402.7SD 6.1% RSD0.1ED. N0: 04Effective Date:APPROVED::HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationED. N0: 04Effective Date:APPROVED::PRECISION - Method RepeatabilityThe full HPLC method as described in SI-IAG-206-02 was carried-out on the finished product IAG-21-001 for the 20mg dosage form. The method repeated six times and the relative standard deviation (RSD) was calculated.SAMPLENumber%ASSAYof labeled amountI 96.9II 97.8III 98.2IV 97.4V 97.7VI 98.5(%) Average97.7SD 0.6(%) RSD0.6PRECISION - Intermediate PrecisionThe full method as described in SI-IAG-206-02 was carried-out on the finished product IAG-21-001 for the 20mg dosage form. The method was repeated six times by a second analyst on a different day using a different HPLC instrument. The average assay and the relative standard deviation (RSD) were calculated.SAMPLENumber% ASSAYof labeled amountI 98.3II 96.3III 94.6IV 96.3V 97.8VI 93.3Average (%)96.1SD 2.0RSD (%)2.1The difference between the average results of method repeatability and the intermediate precision is 1.7%.HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationLINEARITYStandard solutions were prepared at 50% to 200% of the nominal concentration required by the assay procedure. Linear regression analysis demonstrated acceptability of the method for quantitative analysis over the concentration range required. Y-Intercept was found to be insignificant.RANGEDifferent concentrations of the sample (IAG-21-001) for the 20mg dosage form were prepared, covering between 50% - 200% of the nominal weight of the sample.Conc. (%)Conc. (mg/mL)Peak Area% Assayof labeled amount500.20116235096.7700.27935334099.21000.39734463296.61500.64480757797.52000.79448939497.9(%) Average97.6SD 1.0(%) RSD 1.0ED. N0: 04Effective Date:APPROVED::HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationED. N0: 04Effective Date:APPROVED::RANGE (cont.)The results demonstrate linearity as well over the specified range.Correlation coefficient (RSQ)0.99981 Slope11808.3Y -Interceptresponse at 100%* 100 (%) 0.3%ACCURACYACCURACY OF STANDARD INJECTIONSFive (5) replicate injections of the working standard solution at concentration of 0.4242mg/mL, as described in method SI-IAG-206-02 were made.INJECTIONNO.PEAK AREA%ACCURACYI 539299.7II 540599.9III 540499.9IV 5406100.0V 5407100.0Average 5402.899.9%SD 6.10.1RSD, (%)0.10.1The percent deviation from the true value wasdetermined from the linear regression lineHPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationED. N0: 04Effective Date:APPROVED::ACCURACY OF THE DRUG PRODUCTAdmixtures of non-actives (placebo, batch IAG-21-001 ) with Fluoxetine HCl were prepared at the same proportion as in a capsule (70%-180% of the nominal concentration).Three preparations were made for each concentration and the recovery was calculated.Conc.(%)Placebo Wt.(mg)Fluoxetine HCl Wt.(mg)Peak Area%Accuracy Average (%)70%7079.477.843465102.27079.687.873427100.77079.618.013465100.0101.0100%10079.6211.25476397.910080.8011.42491799.610079.6011.42485498.398.6130%13079.7214.90640599.413080.3114.75632899.213081.3314.766402100.399.618079.9920.10863699.318079.3820.45879499.418080.0820.32874899.599.4Placebo, Batch Lot IAG-21-001HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationED. N0: 04Effective Date:APPROVED::VALIDATION OF FLUOXETINE HClAT LOW CONCENTRATIONLINEARITY AT LOW CONCENTRATIONSStandard solution of Fluoxetine were prepared at approximately 0.02%-1.0% of the working concentration required by the method SI-IAG-206-02. Linear regression analysis demonstrated acceptability of the method for quantitative analysis over this range.ACCURACY OF FLUOXETINE HCl AT LOW CONCENTRATIONThe peak areas of the standard solution at the working concentration were measured and the percent deviation from the true value, as determined from the linear regression was calculated.SAMPLECONC.µg/100mLAREA FOUND%ACCURACYI 470.56258499.7II 470.56359098.1III 470.561585101.3IV 470.561940100.7V 470.56252599.8VI 470.56271599.5(%) AverageSlope = 132.7395299.9SD Y-Intercept = -65.872371.1(%) RSD1.1HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationSystem RepeatabilitySix replicate injections of standard solution at 0.02% and 0.05% of working concentration as described in method SI-IAG-206-02 were made and the relative standard deviation was calculated.SAMPLE FLUOXETINE HCl AREA0.02%0.05%I10173623II11503731III10103475IV10623390V10393315VI10953235Average10623462RSD, (%) 5.0 5.4Quantitation Limit - QLThe quantitation limit ( QL) was established by determining the minimum level at which the analyte was quantified. The quantitation limit for Fluoxetine HCl is 0.02% of the working standard concentration with resulting RSD (for six injections) of 5.0%. Detection Limit - DLThe detection limit (DL) was established by determining the minimum level at which the analyte was reliably detected. The detection limit of Fluoxetine HCl is about 0.01% of the working standard concentration.ED. N0: 04Effective Date:APPROVED::HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationED. N0: 04Effective Date:APPROVED::VALIDATION FOR META-FLUOXETINE HCl(EVALUATING POSSIBLE IMPURITIES)Meta-Fluoxetine HCl linearity at 0.05% - 1.0%Relative Response Factor (F)Relative response factor for Meta-Fluoxetine HCl was determined as slope of Fluoxetine HCl divided by the slope of Meta-Fluoxetine HCl from the linearity graphs (analysed at the same time).F =132.7395274.859534= 1.8Detection Limit (DL) for Fluoxetine HClThe detection limit (DL) was established by determining the minimum level at which the analyte was reliably detected.Detection limit for Meta Fluoxetine HCl is about 0.02%.Quantitation Limit (QL) for Meta-Fluoxetine HClThe QL is determined by the analysis of samples with known concentration of Meta-Fluoxetine HCl and by establishing the minimum level at which the Meta-Fluoxetine HCl can be quantified with acceptable accuracy and precision.Six individual preparations of standard and placebo spiked with Meta-Fluoxetine HCl solution to give solution with 0.05% of Meta Fluoxetine HCl, were injected into the HPLC and the recovery was calculated.HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationED. N0: 04Effective Date:APPROVED::META-FLUOXETINE HCl[RECOVERY IN SPIKED SAMPLES].Approx.Conc.(%)Known Conc.(µg/100ml)Area in SpikedSampleFound Conc.(µg/100mL)Recovery (%)0.0521.783326125.735118.10.0521.783326825.821118.50.0521.783292021.55799.00.0521.783324125.490117.00.0521.783287220.96996.30.0521.783328526.030119.5(%) AVERAGE111.4SD The recovery result of 6 samples is between 80%-120%.10.7(%) RSDQL for Meta Fluoxetine HCl is 0.05%.9.6Accuracy for Meta Fluoxetine HClDetermination of Accuracy for Meta-Fluoxetine HCl impurity was assessed using triplicate samples (of the drug product) spiked with known quantities of Meta Fluoxetine HCl impurity at three concentrations levels (namely 80%, 100% and 120% of the specified limit - 0.05%).The results are within specifications:For 0.4% and 0.5% recovery of 85% -115%For 0.6% recovery of 90%-110%HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationED. N0: 04Effective Date:APPROVED::META-FLUOXETINE HCl[RECOVERY IN SPIKED SAMPLES]Approx.Conc.(%)Known Conc.(µg/100mL)Area in spikedSample Found Conc.(µg/100mL)Recovery (%)[0.4%]0.4174.2614283182.66104.820.4174.2614606187.11107.370.4174.2614351183.59105.36[0.5%]0.5217.8317344224.85103.220.5217.8316713216.1599.230.5217.8317341224.81103.20[0.6%]0.6261.3918367238.9591.420.6261.3920606269.81103.220.6261.3920237264.73101.28RECOVERY DATA DETERMINED IN SPIKED SAMPLESHPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationED. N0: 04Effective Date:APPROVED::REPEATABILITYMethod Repeatability - Meta Fluoxetine HClThe full method (as described in SI-IAG-206-02) was carried out on the finished drug product representing lot number IAG-21-001-(1). The HPLC method repeated serially, six times and the relative standard deviation (RSD) was calculated.IAG-21-001 20mg CAPSULES - FLUOXETINESample% Meta Fluoxetine % Meta-Fluoxetine 1 in Spiked Solution10.0260.09520.0270.08630.0320.07740.0300.07450.0240.09060.0280.063AVERAGE (%)0.0280.081SD 0.0030.012RSD, (%)10.314.51NOTE :All results are less than QL (0.05%) therefore spiked samples with 0.05% Meta Fluoxetine HCl were injected.HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationED. N0: 04Effective Date:APPROVED::Intermediate Precision - Meta-Fluoxetine HClThe full method as described in SI-IAG-206-02 was applied on the finished product IAG-21-001-(1) .It was repeated six times, with a different analyst on a different day using a different HPLC instrument.The difference between the average results obtained by the method repeatability and the intermediate precision was less than 30.0%, (11.4% for Meta-Fluoxetine HCl as is and 28.5% for spiked solution).IAG-21-001 20mg - CAPSULES FLUOXETINESample N o:Percentage Meta-fluoxetine% Meta-fluoxetine 1 in spiked solution10.0260.06920.0270.05730.0120.06140.0210.05850.0360.05560.0270.079(%) AVERAGE0.0250.063SD 0.0080.009(%) RSD31.514.51NOTE:All results obtained were well below the QL (0.05%) thus spiked samples slightly greater than 0.05% Meta-Fluoxetine HCl were injected. The RSD at the QL of the spiked solution was 14.5%HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationSPECIFICITY - STABILITY INDICATING EVALUATIONDemonstration of the Stability Indicating parameters of the HPLC assay method [SI-IAG-206-02] for Fluoxetine 10 & 20mg capsules, a suitable photo-diode array detector was incorporated utilizing a commercial chromatography software managing system2, and applied to analyze a range of stressed samples of the finished drug product.GLOSSARY of PEAK PURITY RESULT NOTATION (as reported2):Purity Angle-is a measure of spectral non-homogeneity across a peak, i.e. the weighed average of all spectral contrast angles calculated by comparing all spectra in the integrated peak against the peak apex spectrum.Purity Threshold-is the sum of noise angle3 and solvent angle4. It is the limit of detection of shape differences between two spectra.Match Angle-is a comparison of the spectrum at the peak apex against a library spectrum.Match Threshold-is the sum of the match noise angle3 and match solvent angle4.3Noise Angle-is a measure of spectral non-homogeneity caused by system noise.4Solvent Angle-is a measure of spectral non-homogeneity caused by solvent composition.OVERVIEWT he assay of the main peak in each stressed solution is calculated according to the assay method SI-IAG-206-02, against the Standard Solution, injected on the same day.I f the Purity Angle is smaller than the Purity Threshold and the Match Angle is smaller than the Match Threshold, no significant differences between spectra can be detected. As a result no spectroscopic evidence for co-elution is evident and the peak is considered to be pure.T he stressed condition study indicated that the Fluoxetine peak is free from any appreciable degradation interference under the stressed conditions tested. Observed degradation products peaks were well separated from the main peak.1® PDA-996 Waters™ ; 2[Millennium 2010]ED. N0: 04Effective Date:APPROVED::HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationFORCED DEGRADATION OF FINISHED PRODUCT & STANDARD 1.UNSTRESSED SAMPLE1.1.Sample IAG-21-001 (2) (20mg/capsule) was prepared as stated in SI-IAG-206-02 and injected into the HPLC system. The calculated assay is 98.5%.SAMPLE - UNSTRESSEDFluoxetine:Purity Angle:0.075Match Angle:0.407Purity Threshold:0.142Match Threshold:0.4251.2.Standard solution was prepared as stated in method SI-IAG-206-02 and injected into the HPLC system. The calculated assay is 100.0%.Fluoxetine:Purity Angle:0.078Match Angle:0.379Purity Threshold:0.146Match Threshold:0.4272.ACID HYDROLYSIS2.1.Sample solution of IAG-21-001 (2) (20mg/capsule) was prepared as in method SI-IAG-206-02 : An amount equivalent to 20mg Fluoxetine was weighed into a 50mL volumetric flask. 20mL Diluent was added and the solution sonicated for 10 minutes. 1mL of conc. HCl was added to this solution The solution was allowed to stand for 18 hours, then adjusted to about pH = 5.5 with NaOH 10N, made up to volume with Diluent and injected into the HPLC system after filtration.Fluoxetine peak intensity did NOT decrease. Assay result obtained - 98.8%.SAMPLE- ACID HYDROLYSISFluoxetine peak:Purity Angle:0.055Match Angle:0.143Purity Threshold:0.096Match Threshold:0.3712.2.Standard solution was prepared as in method SI-IAG-206-02 : about 22mg Fluoxetine HCl were weighed into a 50mL volumetric flask. 20mL Diluent were added. 2mL of conc. HCl were added to this solution. The solution was allowed to stand for 18 hours, then adjusted to about pH = 5.5 with NaOH 10N, made up to volume with Diluent and injected into the HPLC system.Fluoxetine peak intensity did NOT decrease. Assay result obtained - 97.2%.ED. N0: 04Effective Date:APPROVED::HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationSTANDARD - ACID HYDROLYSISFluoxetine peak:Purity Angle:0.060Match Angle:0.060Purity Threshold:0.099Match Threshold:0.3713.BASE HYDROLYSIS3.1.Sample solution of IAG-21-001 (2) (20mg/capsule) was prepared as per method SI-IAG-206-02 : An amount equivalent to 20mg Fluoxetine was weight into a 50mL volumetric flask. 20mL Diluent was added and the solution sonicated for 10 minutes. 1mL of 5N NaOH was added to this solution. The solution was allowed to stand for 18 hours, then adjusted to about pH = 5.5 with 5N HCl, made up to volume with Diluent and injected into the HPLC system.Fluoxetine peak intensity did NOT decrease. Assay result obtained - 99.3%.SAMPLE - BASE HYDROLYSISFluoxetine peak:Purity Angle:0.063Match Angle:0.065Purity Threshold:0.099Match Threshold:0.3623.2.Standard stock solution was prepared as per method SI-IAG-206-02 : About 22mg Fluoxetine HCl was weighed into a 50mL volumetric flask. 20mL Diluent was added. 2mL of 5N NaOH was added to this solution. The solution was allowed to stand for 18 hours, then adjusted to about pH=5.5 with 5N HCl, made up to volume with Diluent and injected into the HPLC system.Fluoxetine peak intensity did NOT decrease - 99.5%.STANDARD - BASE HYDROLYSISFluoxetine peak:Purity Angle:0.081Match Angle:0.096Purity Threshold:0.103Match Threshold:0.3634.OXIDATION4.1.Sample solution of IAG-21-001 (2) (20mg/capsule) was prepared as per method SI-IAG-206-02. An equivalent to 20mg Fluoxetine was weighed into a 50mL volumetric flask. 20mL Diluent added and the solution sonicated for 10 minutes.1.0mL of 30% H2O2 was added to the solution and allowed to stand for 5 hours, then made up to volume with Diluent, filtered and injected into HPLC system.Fluoxetine peak intensity decreased to 95.2%.ED. N0: 04Effective Date:APPROVED::HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationSAMPLE - OXIDATIONFluoxetine peak:Purity Angle:0.090Match Angle:0.400Purity Threshold:0.154Match Threshold:0.4294.2.Standard solution was prepared as in method SI-IAG-206-02 : about 22mg Fluoxetine HCl were weighed into a 50mL volumetric flask and 25mL Diluent were added. 2mL of 30% H2O2 were added to this solution which was standing for 5 hours, made up to volume with Diluent and injected into the HPLC system.Fluoxetine peak intensity decreased to 95.8%.STANDARD - OXIDATIONFluoxetine peak:Purity Angle:0.083Match Angle:0.416Purity Threshold:0.153Match Threshold:0.4295.SUNLIGHT5.1.Sample solution of IAG-21-001 (2) (20mg/capsule) was prepared as in method SI-IAG-206-02 . The solution was exposed to 500w/hr. cell sunlight for 1hour. The BST was set to 35°C and the ACT was 45°C. The vials were placed in a horizontal position (4mm vials, National + Septum were used). A Dark control solution was tested. A 2%w/v quinine solution was used as the reference absorbance solution.Fluoxetine peak decreased to 91.2% and the dark control solution showed assay of 97.0%. The difference in the absorbance in the quinine solution is 0.4227AU.Additional peak was observed at RRT of 1.5 (2.7%).The total percent of Fluoxetine peak with the degradation peak is about 93.9%.SAMPLE - SUNLIGHTFluoxetine peak:Purity Angle:0.093Match Angle:0.583Purity Threshold:0.148Match Threshold:0.825 ED. N0: 04Effective Date:APPROVED::HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationSUNLIGHT (Cont.)5.2.Working standard solution was prepared as in method SI-IAG-206-02 . The solution was exposed to 500w/hr. cell sunlight for 1.5 hour. The BST was set to 35°C and the ACT was 42°C. The vials were placed in a horizontal position (4mm vials, National + Septum were used). A Dark control solution was tested. A 2%w/v quinine solution was used as the reference absorbance solution.Fluoxetine peak was decreased to 95.2% and the dark control solution showed assay of 99.5%.The difference in the absorbance in the quinine solution is 0.4227AU.Additional peak were observed at RRT of 1.5 (2.3).The total percent of Fluoxetine peak with the degradation peak is about 97.5%. STANDARD - SUNLIGHTFluoxetine peak:Purity Angle:0.067Match Angle:0.389Purity Threshold:0.134Match Threshold:0.8196.HEAT OF SOLUTION6.1.Sample solution of IAG-21-001-(2) (20 mg/capsule) was prepared as in method SI-IAG-206-02 . Equivalent to 20mg Fluoxetine was weighed into a 50mL volumetric flask. 20mL Diluent was added and the solution was sonicated for 10 minutes and made up to volume with Diluent. 4mL solution was transferred into a suitable crucible, heated at 105°C in an oven for 2 hours. The sample was cooled to ambient temperature, filtered and injected into the HPLC system.Fluoxetine peak was decreased to 93.3%.SAMPLE - HEAT OF SOLUTION [105o C]Fluoxetine peak:Purity Angle:0.062Match Angle:0.460Purity Threshold:0.131Match Threshold:0.8186.2.Standard Working Solution (WS) was prepared under method SI-IAG-206-02 . 4mL of the working solution was transferred into a suitable crucible, placed in an oven at 105°C for 2 hours, cooled to ambient temperature and injected into the HPLC system.Fluoxetine peak intensity did not decrease - 100.5%.ED. N0: 04Effective Date:APPROVED::。
一种司美格鲁肽的合成方法
一种司美格鲁肽的合成方法一种司美格鲁肽的合成方法简介司美格鲁肽（simeprevir）是一种用于治疗丙型肝炎的抗病毒药物。
本文将详细介绍一种合成司美格鲁肽的方法，包括以下几个步骤：1.原料准备2.芳香溴代磺酰胺的合成3.踝基苯并并哌啶的合成4.肽键的形成5.最终产物的提取原料准备合成司美格鲁肽的原料包括： - 芳香溴代磺酰胺 - 踝基苯并并哌啶 - 氯甲烷 - 没食子酸乙酯 - 氯化氢 - 氢氧化钠 - 三氟乙酸 - 氨水芳香溴代磺酰胺的合成1.将没食子酸乙酯加入氯甲烷中，搅拌溶解，得到混合溶液。
2.向混合溶液中滴加氯化氢，在20-25°C下搅拌反应4小时，得到溴代磺酰胺。
踝基苯并并哌啶的合成1.将溴代磺酰胺溶解于二甲基甲酰胺中，得到溶液A。
2.将氨水慢慢滴加到溶液A中，反应1小时，得到中间产物。
3.将中间产物加入氯甲烷中，并加入氢氧化钠调pH值，得到混合溶液。
4.向混合溶液中滴加氯甲烷溶解的三氟乙酸，反应30分钟，得到踝基苯并并哌啶。
肽键的形成1.将踝基苯并并哌啶与氯甲烷中的氯甲酰氯反应，得到酯化产物。
2.将酯化产物溶解于氯甲烷中，加入三氟乙酸，反应30分钟，得到双取代产物。
3.将双取代产物与司美格鲁肽前体反应，得到最终产物。
最终产物的提取将反应溶液中的杂质通过过滤或萃取等方法去除，最后通过结晶、洗涤等步骤得到纯净的司美格鲁肽。
以上即为一种合成司美格鲁肽的详细方法，包括原料准备、芳香溴代磺酰胺的合成、踝基苯并并哌啶的合成、肽键的形成以及最终产物的提取等步骤。
这种方法为司美格鲁肽的大规模合成提供了重要指导。
注意：本文仅为技术讨论，不宣称用于实际操作，任何实际操作需遵循相关法律法规和实验室操作规范。
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］。
WHO International Standard 1st WHO International Standard for Human Papillomavirus (HPV) Type 16 DNA
WHO International Standard1st WHO International Standard for Human Papillomavirus (HPV)Type 16 DNA NIBSC code: 06/202 Instructions for use(Version 2.0, Dated 10/11/2010)1. INTENDED USEThe 1st International Standard for HPV Type 16 (HPV-16) DNA Nucleic Acid Amplification Techniques consists of a freeze-dried preparation of recombinant plasmid containing full-length HPV-16 DNA cloned via its unique BamH1 site (Quint et al., 2006). The standard has been formulated in a background of purified human genomic DNA, lyophilized in 0.5 ml aliquots and stored at -20 °C. The material was calibrated in an international collaborative study involving 19 laboratories (Wilkinson et al., 2008). The International Standard contains material that is proprietory to third parties and should be used for the sole purpose of calibrating in-house or working standards for the amplification and detection of HPV-16 DNA. The International Standard should not be used for any other purpose and should be discarded after use. 2. CAUTIONThis preparation is not for administration to humans .This material contains DNA derived from C33A cells. As with all materials of biological origin, this preparation should be regarded as potentially hazardous to health. It should be used and discarded according to your own laboratory's safety procedures. Such safety procedures should include the wearing of protective gloves and avoiding the generation of aerosols. Care should be exercised in opening ampoules or vials, to avoid cuts.3. UNITAGEThe 1st International Standard for HPV-16 DNA Nucleic Acid Amplification Techniques has been assigned a unitage of 5 x 106 International Units (IU) per ampoule.Traceability statement:It was proposed at a WHO meeting in January 2008 (WHO Meeting Report, 2008) that the instructions for use of the International Standard for HPV-16 DNA include the calculations and assumptions used in determining the theoretical HPV-16 qenome equivalents (GEq) of the bulk material used in formulating the International Standard, thus demonstrating that 1 IU is equivalent to 1 GEq for HPV-16 DNA . The definitive unitage of the 1st WHO International Standard for HPV-16 DNA therefore remains as IU while the traceability statement would allow users to equate IU with GEq.Assays for DNA concentration of the recombinant HPV-16 plasmid stock preparation were performed in Dr Cosette Wheeler‟s laboratory, University of New Mexico (UNM). DNA concentrations were determined by absorbance at 260 nm as well as spectrofluorometrically using the Picogreen assay (Invitrogen Corporation, USA). A correlation coefficient of 0.95 or higher was obtained between the two DNA measurements. 10 ng HPV-16 plasmid DNA/μl was supplied to NIBSC for formulating the bulk material for subsequent freeze-drying. The UNM laboratory also provided NIBSC with a statement indicating that 1.0 x 1011 GEq/ml for HPV-16 is equal to 1.17 ng/μl. 10 ng HPV-16 plasmid DNA/μl plasmid stock preparation is therefore equivalent to 8.547 x 1011 HPV-16 GEq/ml. NIBSC used this data in formulating the 1st International Standard for HPV Type 16 DNA.Formulation of bulk material for the 1st International Standard for HPV Type 16 DNA (NIBSC code 06/202):At NIBSC, the bulk HPV-16 plasmid DNA material was prepared according to the formula:HPV GEq/ml of bulk material = (HPV GEq/ml of plasmid stock x volume plasmid stock) / volume bulk material.Therefore,HPV-16 GEq/ml of bulk material = (8.547 x 1011 HPV-16 GEq/ml plasmid stock) x (0.02223 ml HPV-16 plasmid stock) / 1900 ml HPV-16 bulk material = 1.0 x 107 HPV-16 GEq/ml bulk materialThe HPV-16 DNA bulk material was subsequently freeze-dried in 0.5 ml aliquots.Certain assumptions are required for equating IU to GEq for the 1st International Standard for HPV-16 DNA: 1) 1.0 x 1011 GEq/ml for HPV-16 is equal to 1.17 ng/μl. 2) There is no loss in activity of the HPV-16 DNA upon lyophilization. 3) The recombinant HPV-16 plasmid DNA accurately mimics the activity of HPV-16 viral DNA in biological samples.Independent calculation of GEq/ml for recombinant HPV-16 plasmid DNA.NIBSC also independently calculated the genome equivalence of the HPV-16 plasmid stock preparation and bulk preparation in which the molecular weights of the full-length HPV-16 genome and pBR322 DNA were based on sequence content using BioEdit Sequence Alignment Editor v7.0.5.3 (Tom Hall, Isis Pharmaceuticals Inc., USA). The sequences used for determining the molecular weights are GenBank Accession number J01749.1 for pBR322 and the reference sequence for HPV16 (Accession K02718).BioEdit dataDNA molecule: HPV16 Accession K02718 Length = 7904 base pairsMW= 4786756.00 Daltons, double strandedDNA molecule: cloning vector pBR322 Length = 4361 base pairsMW= 2653867.00 Daltons, double strandedFormulaeGEq/ml of the HPV plasmid stock was calculated according to the formula: GEq/ml of the HPV plasmid stock = (DNA concentration of HPV plasmid stock) x (MW of HPV DNA + MW of pBR322)-1 x (Avogadro‟s Number) where Avogadro‟s Number = 6.022x1023 molecules/molGEq/ml of the bulk HPV DNA materials was calculated according to the formula:HPV GEq/ml of bulk material = (HPV GEq/ml of plasmid stock x volume plasmid stock) / volume bulk material.CalculationThe recombinant HPV-16 plasmid stock preparation was supplied to NIBSC at a concentration of 10 ng/μl. Using the MW determinations shown above, the GEq/ml of the HPV-16 plasmid stock is:= (10 x 10-9 g/μl) x (mol/(7440623 g) x (6.022x1023 molecules/mol) = 8.093 x 108 molecules/μl = 8.093 x 1011molecules/ml = 8.093 x 1011 HPV-16 GEq/ml22.23μl of the recombinant HPV-16 plasmid stock was diluted to a final volume of 1900ml, therefore,HPV-16 GEq/ml of bulk material = (8.093 x 1011 HPV-16 GEq/ml plasmid stock) x (0.02223 ml HPV-16 plasmid stock) / 1900 ml HPV-16 bulk material = 0.947 x 107 HPV-16 GEq/ml bulk material4. CONTENTSCountry of origin of biological material: United Kingdom.Each ampoule contains the lyophilized equivalent of 0.5 ml HPV-16 plasmid DNA in 10mM Tris buffer pH7.4 containing 1mM EDTA, 5 mg/ml trehalose and ~1 x 106 human GEq/ml derived from C33a cells.5. STORAGEThe ampoule should be stored at -20 °C or below on receipt.Please note: because of the inherent stability of lyophilized material, NIBSC may ship these materials at ambient temperature.6. DIRECTIONS FOR OPENINGDIN ampoules have an …easy -open‟ coloured stress point, where the narrow ampoule stem joins the wider ampoule body.Tap the ampoule gently to collect the material at the bottom (labeled) end. Ensure that the disposable ampoule safety breaker provided is pushed down on the stem of the ampoule and against the shoulder of the ampoule body. Hold the body of the ampoule in one hand and the disposable ampoule breaker covering the ampoule stem between the thumb and first finger of the other hand. Apply a bending force to open the ampoule at the coloured stress point, primarily using the hand holding the plastic collar.Care should be taken to avoid cuts and projectile glass fragments that might enter the eyes, for example, by the use of suitable gloves and an eye shield. Take care that no material is lost from the ampoule and no glass falls into the ampoule. Within the ampoule is dry nitrogen gas at slightly less than atmospheric pressure. A new disposable ampoule breaker is provided with each DIN ampoule.7. USE OF MATERIALNo attempt should be made to weigh out any portion of the freeze-dried material prior to reconstitution.The 1st International Standard for HPV-16 DNA contains high copy number template. There is a high risk of HPV-16 plasmid DNA contamination via aerosolization upon opening of the glass ampoule. The material must be opened and handled in a separate laboratory environment, away from other pre-amplification components such as reagents, labware and samples.The material is supplied lyophilized and, before use, should be reconstituted in 0.5 ml sterile nuclease-free water. Ensure that the inside surface of the ampoule is wetted with the added water so that any particles of freeze-dried material adhering to the glass are reconstituted. The reconstituted material has a final concentration of 1 X 107 IU/ml. The reconstituted material is suitable for calibration of in-house or working standards for the amplification and detection of HPV-16 DNA.. The material is not suitable for calibrating or assessing extraction, precipitation or centrifugation procedures. The material has NOT been calibrated for human DNA nucleic acid amplification techniques.8. STABILITYReference materials are held at NIBSC within assured, temperature-controlled storage facilities. The 1st International Standard for HPV-16 DNA should be stored at -20 °C or below on receipt.Studies on the stability of reconstituted standard are underway. Users should determine the stability of the reconstituted material according to their own method of preparation, storage and use.NIBSC follows the policy of WHO with respect to its reference materials.9. REFERENCESQuint, W. G. V., Pagliusi, S. R., Lelie, N., de Villiers, E. M., Wheeler, C. M. and the World Health Organization Human Papillomavirus DNA International Collaborative Study Group. (2006). Results of the First WorldHealth Organization International Collaborative Study of Detection of Human Papillomavirus DNA. J. Clin. Microbiol. 44: 571-579.Wilkinson, D.E., Baylis, S.A., Padley, D., Heath, A.B., Ferguson, M., Pagliusi, S.R., et al. Establishment of the 1st World Health Organization international standards for human papillomavirus type 16 DNA and type 18 DNA. Int J Cancer 2010 Jun 15;126(12):2969-83.WHO meeting report, on “Standardization of HPV assays and the role of HPV LabNet in supporting vaccine introduction” Geneva, Switzerland, 23-25 January 2008, in preparation.10. ACKNOWLEDGEMENTS11. FURTHER INFORMATIONFurther information can be obtained as follows; This material: enquiries@ WHO Biological Standards:http://www.who.int/biologicals/en/JCTLM Higher order reference materials: /en/committees/jc/jctlm/ Derivation of International Units:/products/biological_reference_materials/frequently _asked_questions/how_are_international_units.aspx Ordering standards from NIBSC:/products/ordering_information/frequently_asked_q uestions.aspxNIBSC Terms & Conditions:/terms_and_conditions.aspx12. CUSTOMER FEEDBACKCustomers are encouraged to provide feedback on the suitability or use of the material provided or other aspects of our service. Please send any comments to enquiries@13. CITATIONIn all publications, including data sheets, in which this material is referenced, it is important that the preparation's title, its status, the NIBSC code number, and the name and address of NIBSC are cited and cited correctly.15. LIABILITY AND LOSSInformation provided by the Institute is given after the exercise of all reasonable care and skill in its compilation, preparation and issue, but it is provided without liability to the Recipient in its application and use. It is the responsibility of the Recipient to determine the appropriateness of the standards or reference materials supplied by the Institute to the Recip ient (“the Goods”) for the proposed application and ensure that it has the necessary technical skills to determine that they are appropriate. Results obtained from the Goods are likely to be dependant on conditions of use by the Recipient and the variability of materials beyond the control of the Institute.All warranties are excluded to the fullest extent permitted by law, including without limitation that the Goods are free from infectious agents or that the supply of Goods will not infringe any rights of any third party.The Institute shall not be liable to the Recipient for any economic loss whether direct or indirect, which arise in connection with this agreement.The total liability of the Institute in connection with this agreement, whether for negligence or breach of contract or otherwise, shall in no event exceed 120% of any price paid or payable by the Recipient for the supply of the Goods.If any of the Goods supplied by the Institute should prove not to meet their specification when stored and used correctly (and provided that the Recipient has returned the Goods to the Institute together with written notification of such alleged defect within seven days of the time when the Recipient discovers or ought to have discovered the defect), the Institute shall either replace the Goods or, at its sole option, refund the handling charge provided that performance of either one of the above options shall constitute an entire discharge of the Institute‟s liability under this Condition.。
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。
不同部位脑梗死华勒变性的特点及对神经功能的影响
［8］涂梦恬，李良平.肝细胞核因子受体4α在非酒精性脂肪性肝病中的研究现状［J］.国际消化病杂志，2017，37（3）：144-147.Tu MT，Li LP.Research status of hepatocyte nuclear factor receptor4αin nonalcoholic fatty liver disease［J］.Int J Gastroenterol，2017，37（3）：144-147.doi：10.3969/j.issn.1673-534X.2017.03.003.［9］Thayer TE，Lino Cardenas CL，Martyn T，et al.The role of bone morphogenetic protein signaling in non-alcoholic fatty liver disease ［J］.Sci Rep，2020，10（1）：9831.doi：10.1038/s41598-020-66770-8.［10］Cariou B，Byrne CD，Loomba R，et al.NAFLD as a metabolic disease in humans：A literature review［J］.Diabetes Obes Metab，2021，23（5）：1069-1083.doi：10.1111/dom.14322.［11］Theofilatos D，Anestis A，Hashimoto K，et al.Transcriptional regulation of the human liver X receptorαgene by hepatocyte nuclear factor4α［J］.Biochem Biophys Res Commun，2016，469（3）：573-579.doi：10.1016/j.bbrc.2015.12.031.［12］Vespasiani-Gentilucci U，Dell'Unto C，De Vincentis A，et al. Combining genetic variants to improve risk prediction for NAFLD and its progression to cirrhosis：a proof of concept study［J］.Can J Gastroenterol Hepatol，2018，2018：7564835.doi：10.1155/2018/ 7564835.［13］Khalid YS，Dasu NR，Suga H，et al.Increased cardiovascular events and mortality in females with NAFLD：a meta-analysis［J］. Am J Cardiovasc Dis，2020，10（3）：258-271.［14］Trépo E，Valenti L.Update on NAFLD genetics：From new variants to the clinic［J］.J Hepatol，2020，72（6）：1196-1209.doi：10.1016/ j.jhep.2020.02.020.［15］Matsuo S，Ogawa M，Muckenthaler MU，et al.Hepatocyte nuclear factor4alpha controls iron metabolism and regulates transferrin receptor2in mouse liver［J］.J Biol Chem，2015，290（52）：30855-30865.doi：10.1074/jbc.M115.694414.［16］Chi X，Wei X，Gao W，et al.Dexmedetomidine ameliorates acute lung injury following orthotopic autologous liver transplantation in rats probably by inhibiting Toll-like receptor4-nuclear factor kappa B signaling［J］.J Transl Med，2015，13：190.doi：10.1186/ s12967-015-0554-5.［17］Wang L，Yao M，Zheng W，et al.Insulin-like growth factor I receptor：a novel target for hepatocellular carcinoma gene therapy ［J］.Mini Rev Med Chem，2019，19（4）：272-280.doi：10.2174/ 1389557518666181025151608.（2020-10-23收稿2021-01-17修回）（本文编辑李鹏）不同部位脑梗死华勒变性的特点及对神经功能的影响王冬梅，蔡桂淑，刘立生△摘要：目的探讨不同部位脑梗死华勒变性的特点及对神经功能的影响。
细胞蛇的研究进展
2007年,英国牛津大学的刘骥陇等在研究果蝇U 小体和P 小体(U 小体和P 小体是真核生物细胞质中的无膜细胞器)的功能关系时,用4种针对Cup (P 小体中的一种蛋白质)的抗体,对雌性果蝇的卵巢组织进行免疫组织化学染色,染色结果除了预期标记上的P 小体外,还标记出了长条形的丝状结构[1]。
这种结构的形状和数量与纤毛很相似,导致当时以为在果蝇中找到了有纤毛的新细胞类型。
但后来的一系列实验表明,该结构与纤毛没有关系,于是将其命名为“细胞蛇”。
最初是抗Cup 抗体不纯产生假象,意外发现的细胞蛇,而采用亲和层析纯化后的抗Cup 抗体无法再DOI:10.16605/ki.1007-7847.2020.10.0258细胞蛇的研究进展收稿日期:2020-10-22;修回日期:2020-11-19;网络首发日期:2021-07-27基金项目:宁夏自然科学基金项目(2020AAC03179);国家自然科学基金资助项目(31560329)作者简介:李欣玲(1999—),女,广西贵港人,学生;*通信作者:俞晓丽(1984—),女,宁夏银川人,博士,副教授,主要从事干细胞与生殖生物学研究,E-mail:********************。
李欣玲,张樱馨,李进兰,潘文鑫,王彦凤,杨丽蓉,王通,俞晓丽*(宁夏医科大学生育力保持教育部重点实验室临床医学院基础医学院,中国宁夏银川750000)摘要:细胞蛇是近年来细胞生物学研究的热门方向之一,由于其在细胞的增殖、代谢和发育上具有一定的生物学功能,因此,对一些疾病如癌症等的临床诊断或治疗具有一定的指导意义。
细胞蛇是由三磷酸胞苷合成酶(cytidine triphosphate synthetase,CTPS)聚合而成的无膜细胞器,其形成过程及功能在不同类型的细胞中不尽相同。
例如:细胞蛇能促进癌细胞增殖,并使患者病情恶化;过表达的细胞蛇可抑制神经干细胞增殖,影响大脑皮层发育;在卵泡细胞中,细胞蛇相当于CTPS 的存储库,在卵子发生过程起到促进细胞增殖和代谢的作用。
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。
异丁香酚的离体透皮吸收研究
异丁香酚的离体透皮吸收研究
周梅华;刘静波
【期刊名称】《中国公共卫生》
【年(卷),期】2000(16)10
【摘要】采用静态渗透室装置 ,测定异丁香酚皮片涂药后不同时间对小鼠离体皮肤透皮吸收率 ,异丁香酚采用GC法测定 ,结果 :异丁香酚经离体小鼠皮肤的渗透率随时间的增加而增加 ,2 4h后 ,异丁香酚的渗透率为 1 0 0 %。
结论
【总页数】1页(P907-907)
【关键词】异丁香酚;气相色谱;离体透皮吸收
【作者】周梅华;刘静波
【作者单位】上海医科大学分析化学教研室;上海医科大学预防医学研究所
【正文语种】中文
【中图分类】R944.9
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胰岛素固体脂质纳米粒粉雾剂的制备
胰岛素固体脂质纳米粒粉雾剂的制备毕茹【摘要】目的制备胰岛素固体脂质纳米粒肺吸入粉雾剂.方法复乳法可提高水溶性大分子药物胰岛素的包封率,再用喷雾冷冻干燥技术将混悬液微粉化,考察各因素对粉体性质的影响.结果所得粉末疏松多孔、密度小,具有良好的雾化性能;复溶后仍可得形态圆整的纳米粒,理化性质略有改变.结论通过摸索处方及工艺,制得的粉雾剂既可满足在肺泡上皮区域沉积的要求,又可使纳米粒维持一定的包封率,以达到肺内缓慢释药的目的.【期刊名称】《西北药学杂志》【年(卷),期】2011(026)003【总页数】3页(P204-206)【关键词】胰岛素;固体脂质纳米粒;粉雾剂【作者】毕茹【作者单位】天津国际SOS紧急救援诊所药剂科,天津,300074【正文语种】中文【中图分类】R944近年来蛋白质多肽类大分子药物肺部给药得到广泛研究乃至商品化[1]，肺吸入胰岛素粉雾剂ExuberaTM的上市更成为糖尿病治疗史上的重大突破，但该药只起到速效作用。
笔者对胰岛素肺部给药的长效制剂进行了初步研究。
固体脂质纳米粒(SLN)作为新型的亚微粒给药系统，可延长药物在局部的滞留时间、控制药物的释放、提高生物利用度，具有耐受性好、安全性高等特点[2]，因此特别适用于肺内控释给药。
笔者将胰岛素(INS)包裹于固体脂质纳米粒(INS-SLN)中，再将其通过一定的微粉化方法制备成吸入粉雾剂(INS-SLN-DPI)，以达到缓释长效及提高制剂稳定性的目的。
1.1 仪器高效液相色谱仪(LC-10ATVP，日本岛津)；紫外检测器(SPD-10A VP，日本岛津)；高速冷冻离心机(TGL-16G-A，上海安亭科学仪器厂)；透射电子显微镜(日本电子公司)；粒度分布与电势分析仪(ZETASIZER3000，英国Malvern公司)；喷枪(瑞士Buchi公司 B-191喷雾干燥仪)；冷冻干燥机(军事医学科学院实验仪器厂)；光学显微镜(E200，日本尼康公司)；扫描电子显微镜(S-570，日本日立公司)；胶囊型粉雾吸入器(上海市天平制药厂)；水分测定仪(MB45，美国奥豪斯公司)；有效部位沉积量测定装置(自制)。