Hepatitis C Virus Experimental Model Systems and Antiviral drug Research

An estimated 130 million people worldwide are chronically infected with hepatitis C virus (HCV) making it a leading cause of liver disease worldwide. Because the currently available therapy of pegylated interferon-alpha and ribavirin is only effective in a subset of patients, the development of new HCV antivirals is a healthcare imperative. This review discusses the experimental models available for HCV antiviral drug research, recent advances in HCV antiviral drug development, as well as active research being pursued to facilitate development of new HCV-specific therapeutics.

Regulating effect of Chinese herbal medicine on the peritoneal lymphatic stomata in enhancing ascites absorption of experimental hepatofibrotic mice

AIM: To observe the regulatory effect of Chinese herbal medicine on peritoneal lymphatic stomata and its significance in treating ascites in liver fibrosis model mice. METHODS: Two Chinese herbal composite prescriptions were used separately to treat the carbon tetrachloride-induced mouse model of liver fibrosis. The histo-pathologic changes of the liver sections (HE and VG stainings) were observed. The peritoneal lymphatic stomata was detected by scanning electron microscopy and computer image processing. The changes of urinary volume and sodium ion concentration were measured. RESULTS: In the model group, lots of fibrous tissue formed in liver and extended into the hepatic lobules to separate them incompletely. In the treated and prevention groups, the histo-pathologic changes of liver was rather milder, only showed much less fibrous tissue proliferation in the hepatic lobules. The peritoneal lymphatic stomata enlarged with increased density in the experimental groups (diameter: PA, 3.07 +/- 0.69 microm; PB, 2.82 +/- 0.37 microm; TA, 3.25 +/- 0.82 microm and TB, 2.82 +/- 0.56 microm; density: PA, 7.11 +/- 1.90 stomata.1000 microm(-2); PB, 8.76 +/- 1.45 stomata.1000 microm(-2); TA, 6.55 +/- 1.44 stomata.1000 microm(-2)and TB, 8.76+/-1.79 stomata.1000 microm(-2)), as compared with the model group (diameter: 2.00+/-0.52 microm density: 4.45+/-1.05 stomata.1000 microm(-2)). After treatment, the urinary volume and sodium ion excretion increased in the experimental groups (PA, 231.28+/-41.09 mmol.L(-1); PB, 171.69 +/- 27.48 mmol.L(-1) and TA, 231.44 +/- 34.12 mmol.L(-1)), which were significantly different with those in the model group (129.33 +/- 36.75 mmol.L(-1)). CONCLUSION: Chinese herbal medicine has marked effects in alleviating liver fibrosis, regulating peritoneal lymphatic stomata, improving the drainage of ascites from peritoneal cavity and causing increase of urinary volume and sodium ion excretion to reduce the water and sodium retention, and thus have favorable therapeutic effect in treating ascites.

Bioinformatics screening of breast cancer-related genes and potential drug research

Objective:To search the the differentially expressed genes between breast cell carcinoma tissues and normal tissues by using bioinformatics technology,and the potential therapeutic drugs for breast cancer were identified,which can provide reference for clinical immune targeted therapy and drug therapy of breast cancer in the future.Methods:"Breast cancer"was searched by using Gene Expression Omnibus(GEO),and GSE79586 chip data was downloaded.The differentially expressed genes in the control group and the breast cancer model group were screened by using bio-communication technology and subjected to GO function analysis,KEGG pathway analysis,differential gene characteristic expression analysis and protein-protein interaction network(PPI)analysis,and the analysis results were further visualized.Prognosis analysis,related function prediction and immune infiltration analysis were performed using the GEPIA,GeneMANIA,and Timer2.0 databases,respectively.Finally,the compounds with potential therapeutic effects on breast cancer are identified through Connectivity Map(CMap).Western blotting and real-time PCR(RT-PCR)were used to verify the core genes and potential therapeutic agents with the highest correlation in vitro.Results:A total of 3916 differentially expressed genes including 1786 up-regulated genes and 2130 down-regulated genes were screened.GO analysis showed that the differential genes were mainly involved in the positive regulation of phosphorylation,secretory vesicles,racemase and epimerase activities.KEGG analysis showed that differential genes were involved in systemic lupus erythematosus,alcoholism,sticky spots,amoebic dysentery Ras signal pathways and other disease pathways.The characteristic expression analysis of differential genes showed that MEK inhibitors,HSP90 inhibitors and signal transduction pathway kinase inhibitors were drugs similar to the differential genes.PPI results showed that H2AFJ,TFF1,GATA3,FOXA1,and CDH1 were core genes related to breast cancer.Two core genes of H2AFJ and TFF1 with the highest correlation were further selected for GEPIA analysis.The results of the analysis showed that the mRNA expression levels of H2AFJ and TFF1 in breast cancer cells were significantly higher than those in normal tissues,and there was a significant correlation with the pathological staging,overall survival rate and disease-free survival rate of breast cancer patients.H2AFJ and TFF1 may be potential prognostic biomarkers for survival of breast cancer patients.The functions of differentially expressed H2AFJ and TFF1 are mainly related to hormone receptor binding,epithelial structure maintenance and epigenetic negative regulation of genes,chromatin tissue involved in negative regulation of transcription,etc.The results of immune infiltration showed that the expressions of H2AFJ and TFF1 had a significant correlation with the infiltration of macrophages,neutrophils,monocytes,CD4+T,CD8+T,B lymphocytes and other immune cells.CMap results showed that compounds such as Gefitinib,Alpelisib,Sorafenib,and Sunitinib had potential therapeutic effects on breast cancer.Western blot and RT-PCR results showed that H2AFJ and TFF1 were significantly overexpressed in breast cancer cells.Gefitinib significantly inhibited the expression of H2AFJ and TFF1 in breast cancer cells(P<0.05,P<0.01).Conclusion:In this study,differentially expressed genes between breast cell carcinoma tissues and normal tissues were screened out by bioinformatics means to further identify key genes and compounds with potential therapeutic effects in the onset process of breast cancer and to further verify the effectiveness of the screened drugs on breast cancer through experiments.It will provide reference for clinical research and development of new drugs against breast cancer in the future in order to develop more effective treatment options.

以药物开发为中心的药学综合设计性实验课程设计

实验教学在培养药学本科生理论知识与实践能力上发挥着关键作用。在基于地方高校“双一流”建设的新形势需求下,探索以药学本科教育为核心的实验教学新方法和新观念尤为重要。药物研发是药学专业人才培养的主要目标,是综合运用药理学、药物化学、药剂学、药物分析等学科知识进行再生产的过程,大多数学生也将从事与药物研发、生产相关的职业。设计药物研发全过程的综合设计性实验,对课程的设计、实施、质控和存在的问题及解决办法进行了探讨,为培养高素质、创新型药学人才提供了参考。

《治疗药物监测》实验课线上教学模式的探索

随着“互联网+”时代的到来以及移动终端的普及,传统的实验课线下教学模式呈现出一定的短板,线上教学模式成为课程教学体系的重要组成部分。文章以新疆医科大学第一附属医院临床药学专业必修课程《治疗药物监测》实验课“高效液相色谱技术测定卡马西平血药浓度”为例,围绕临床药学专业人才培养目标和课程体系标准,基于线上平台,从课前准备、课中实施、课后巩固三阶段探索实验课线上教学模式,推动了信息技术与课堂的融合发展,激发学生学习积极主动性,促进了传统教学模式的改革,为高校线上线下混合式实验教学提供一定的经验借鉴和参考。

蛋白质结构稳定性检测的实验课程设计

随着生物技术的突破和发展,多肽蛋白类药物的出现使得人类疾病的诊断和治疗发生了翻天覆地的变化。为了顺应多肽蛋白类药物的飞速发展,加强生命科学等专业本科生对此类药物制剂相关知识的普及已势在必行。此实验课程设计利用稳定性热检测法分析蛋白质结构的稳定性,为生命科学等专业本科生设计一堂深入了解蛋白质结构与功能关系的课程,以促进本科生拓宽学术视野,提升完成相关科学研究和适应医药产业发展新形势的能力。

论药品专利补充实验数据的接受标准

药品专利由于研发时间长,在申请日后会产生大量的补充实验数据。我国的立法及实践已明确,应当对补充实验数据进行审查。但审查不等于被接受,典型案例反映出,对补充实验数据的接受标准依然存在较大争议。域外美国、欧洲、日本的立法模式和司法实践也各不相同。解决当前药品专利补充实验数据接收标准所存在的问题,应当从以下几个方面进行着手:分别规定“技术方案”和“预测效果”不同的接受标准;厘清补充实验数据与证明目的之间的关系;放宽对预测效果的严格标准;广泛理解本领域公知常识及现有技术。

某机构临床试验用药品管理的现存问题及改进措施

目的提升临床试验用药品管理水平,保证临床试验的质量。方法调取2021年3月~2023年7月某机构承接的32个在研项目的试验用药品管理的质控结果,对发现的问题进行归纳分析,并提出改进措施和建议。结果汇总不合格项25项,共计57个问题。通过总结分析各环节出现的问题,从人员、管理、软硬件等方面找出试验用药品管理不规范的原因。提出了完善试验用药品的管理流程,增强试验用药品标准操作规程(SOP)和管理制度的可操作性,加强研究人员的培训,加强受试者的用药教育,建立试验用药品的信息化管理系统,对试验用药品进行定期质量管理的改进措施。结论药物临床试验机构应加强试验用药品的管理和培训,通过建立《药物临床试验质量管理规范》(GCP)中心药房,配备专职药师,提高试验用药品管理水平,保证药物临床试验过程真实规范、结果科学可靠。

基于生物信息学和体外实验验证的肾康注射液治疗肾纤维化作用机制研究

目的 利用生物信息学及体外实验探讨肾康注射液(Shenkang injection, SKI)治疗肾纤维化(renal fibrosis, RF)的作用机制与物质基础。方法 利用GEO数据库筛选RF差异表达基因,借助CMAP数据库,基于基因表达谱相似性原理,重定位出具有调控RF作用的药物,然后通过分子指纹相似性分析筛选出SKI潜在治疗RF的成分;同时基于网络药理学预测SKI调控RF的核心靶点及通路;最后,通过分子对接和细胞实验进行验证。结果 基于GEO数据库筛选到2个RF相关的数据集,CMAP重定位到3个共同的RF治疗药物(saracatinib、dasatinib、PP-2),分子指纹相似性分析发现,RF治疗药物与salvianolic acid B、hydroxysafflor yellow A等5个SKI成分的结构相似性较高。分子对接结果发现,salvianolic acid B、hydroxysafflor yellow A等成分与SKI调控的潜在治疗RF的核心靶标MMP1、MMP13等均有良好的结合能力。网络药理学分析提示,SKI核心靶点主要富集在Relaxin和AGE-RAGE等信号通路。细胞实验表明,SKI可显著降低RF模型细胞AGE-RAGE信号通路中AGER、NFKB1、COL1A1、SERPINE1、VEGFC和Relaxin信号通路中MMP1、MMP13的mRNA表达水平,显著升高RXFP1的mRNA表达水平。结论 SKI可通过调控Relaxin和AGE-RAGE信号通路来发挥治疗RF的作用,其物质基础可能为salvianolic acid B、hydroxysafflor yellow A等成分。

药物分析实验教学设计——以“高效液相法测定阿司匹林肠溶片含量”为例

药物分析是具有很强实践性的应用学科,以学生掌握常用药品的分析方法和实验技术为教学目标。依据《中国药典》,文章选出较为典型的阿司匹林肠溶片为例,设计其含量测定为实验教学内容。详细阐述了实验教学过程设计、教学内容、效果评估与改进。课程教学设计包括了药物分析课程原理、方法及实验技术,形成了完整的实验教学设计、教学效果评估和持续改进的实验教学设计过程和反馈机制。基于以上设计,提高了学生掌握常规药物分析的基本操作技能,培养了学生独立分析解决问题的能力,为提高药物分析实验教学质量奠定坚实基础。

Multi-omics Approach Reveals Influenza-A Virus Target Genes Associated Genomic,Clinical and Immunological Characteristics in Cancers

Objective To examine the precise function of influenza A virus target genes(IATGs)in malignancy.Methods Using multi-omics data from the TCGA and TCPA datasets,33 tumor types were evaluated for IATGs.IATG expression in cancer cells was analyzed using transcriptome analysis.Copy number variation(CNV)was assessed using GISTICS 2.0.Spearman’s analysis was used to correlate mRNA expression with methylation levels.GSEA was used for the enrichment analysis.Pearson’s correlation analysis was used to examine the association between IATG mRNA expression and IC50.The ImmuCellAI algorithm was used to calculate the infiltration scores of 24 immune cell types.Results In 13 solid tumors,IATG mRNA levels were atypically expressed.Except for UCS,UVM,KICH,PCPG,THCA,CHOL,LAMI,and MESO,most cancers contained somatic IATG mutations.The main types of CNVs in IATGs are heterozygous amplifications and deletions.In most tumors,IATG mRNA expression is adversely associated with methylation.RT-PCR demonstrated that EGFR,ANXA5,CACNA1C,CD209,UVRAG were upregulated and CLEC4M was downregulated in KIRC cell lines,consistent with the TCGA and GTEx data.Conclusion Genomic changes and clinical characteristics of IATGs were identified,which may offer fresh perspectives linking the influenza A virus to cancer.

应用植入式缓释泵实现大鼠脑室给药的操作方法及其改进

目的介绍应用植入式缓释泵实现脑室给药的实验技术并予以改进。方法选择8周龄雄性SD大鼠,准备所需器材及试剂。首先将缓释泵组装并孵育至工作状态,然后进行植入手术操作。实验动物麻醉后备皮,手术暴露颅骨上表面,使用脑立体定位仪定位脑室正上方一点,在该点用高速颅钻钻1个小孔。先将泵体埋置于颈部皮下,然后将针头插入小孔中,并用牙科水泥固定,凝固后剪去针头基座,逐层缝合皮下软组织和头皮,将动物放回笼内单独饲养。结果缓释泵泵体成功植入大鼠颈部皮下,针头牢固固定于颅骨,导管接口未断开。取出完整脑组织检查,可见穿刺点及针道周围无明显血肿,脑室内及周围组织可见蓝色染料,表明本方法能够成功将药物送达脑室。结论通过引入脑立体定位仪辅助定位,并对此技术的操作流程进行改进,使植入操作更加精准、安全,具有较高的脑室给药成功率。

禁毒技术创新人才培养初探——以浙江警察学院“毒品防控技术研究人才实验班”为例

禁毒技术创新人才是指适应新时代公安禁毒工作,具有强烈的事业心和社会责任感,富有创新精神和能力,熟练掌握现代仪器设备的禁毒技术人员。该文以浙江警察学院“毒品防控技术研究人才实验班”为例,从培养方案、选拔机制、培养模式、课程设置等多个方面探讨禁毒技术创新人才培养的新路径,相关研究为培养公安院校技术类创新人才提供借鉴。

天麻-丹参药对治疗高血压作用机制的网络药理学分析及实验验证

目的基于网络药理学方法及动物实验验证探讨天麻-丹参药对治疗高血压的作用机制。方法(1)通过TCMSP、BATMAN及TCMIP数据库筛选天麻-丹参药对活性成分及其作用靶点;通过Drugbank、Genecard、TTD、Disgenet数据库检索获得高血压疾病相关靶点;对药对的活性成分作用靶点与高血压疾病相关靶点取交集(共同靶点),所得交集靶点即为天麻-丹参药对治疗高血压的潜在作用靶点。将天麻-丹参药对活性成分及其作用靶点导入Cytoscape 3.9.1软件,构建“中药-活性成分-靶点”网络,筛选关键活性成分;构建潜在作用靶点的蛋白互作(PPI)网络,筛选潜在核心靶点;使用Metascape平台对潜在作用靶点进行GO功能及KEGG通路富集分析。选择关键活性成分与潜在核心靶点进行分子对接验证。(2)将30只雄性自发性高血压大鼠(SHR)随机分为模型组、西药组(坎地沙坦酯,0.72 mg·kg^(-1))及天麻-丹参药对低、中、高剂量组(2.25、4.50、9.00 g·kg^(-1)),另选雄性WKY大鼠为空白组,每组6只,每日1次,连续灌胃给药8周。分别在给药前及药物干预2、4、6、8周后检测大鼠尾动脉收缩压;采用HE染色法观察胸主动脉组织病理变化;Western Blot法检测腹主动脉中GRP78、CHOP、Caspase-12蛋白表达水平。结果(1)共得到天麻-丹参药对活性成分83个,筛选出天麻-丹参药对治疗高血压的潜在作用靶点(交集靶点)158个;5个关键活性成分:对羟基苯甲酸、4-羟基苄胺、丹参酮Ⅰ、丹参酮、γ-谷甾醇;6个潜在核心靶点:IL6、TNF、CASP3、JUN、PTGS2、IL1B;GO功能富集分析得到1826条生物学过程条目、89条细胞组分条目、199条分子功能条目;KEGG通路富集分析获得186条通路,主要涉及神经活性配体-受体相互作用、钙信号通路、炎症应答(如TNF和MAPK信号通路)、血管保护(如HIF-1和cAMP信号通路)、氧化应激(如PI3K-Akt信号通路)等信号通路;丹参酮Ⅰ、丹参酮对6个潜在核心靶点均有较强的结合力,γ-谷甾醇对IL6、CASP3、JUN、PTGS2、IL1B具有较强的结合力。(2)与空白组比较,模型组大鼠的收缩压显著升高(P<0.01);胸主动脉内皮损伤明显,内皮细胞形态异常,可见肿胀、脱落细胞,组织内膜排序紊乱,内膜结构不完整,出现内膜增厚;腹主动脉中GRP78、CHOP、Caspase-12蛋白表达量显著升高(P<0.01)。与模型组比较,给药组大鼠的收缩压均显著下降(P<0.01);胸主动脉损伤减轻,内皮细胞形态、内膜结构及厚度等均得到不同程度改善;腹主动脉中GRP78、CHOP、Caspase-12蛋白表达量显著降低(P<0.01)。结论天麻-丹参药对可能是通过对羟基苯甲酸、丹参酮、γ-谷甾醇等关键活性成分,作用于IL6、TNF、CASP3、JUN、PTGS2、IL1B等核心靶点,调控TNF信号通路、MAPK信号通路、PI3K-Akt信号通路、PERK信号通路等关键信号通路,发挥改善血管内皮功能障碍、抑制内质网应激及降血压的作用。

两种测定阿司匹林肠溶片含量的方法在教学应用中的比较

测定阿司匹林肠溶片含量是药物分析实验的核心教学内容。两步酸碱滴定法和高效液相色谱法是教学实验中最常用的两种方法,本研究从实验原理、方法、实验设施和教学实践能力等多个方面进行比较。两步酸碱滴定法具有实验操作容易、所需实验设备较少,但测定结果不够准确的特点。高效液相色谱法测定结果准确,且可以提升学生使用高效液相色谱的能力。但其实验过程复杂,对实验场地和经费具有较高要求。最后提出以两步酸碱滴定法为基础教学内容,将高效液相色谱法作为拓展内容。在理解阿司匹林肠溶片含量测定原理和方法同时,提升学生应用现代药物分析设备的能力,有助于提高人才培养水平。

BOPPPS教学模式在微生物学与免疫学实验中的应用

微生物学与免疫学是医学院校多个专业的基础必修课,而微生物学与免疫学实验课程作为理论课的延伸,在促进理论知识掌握的同时,也锻炼了学生的专业实践能力,是培养创新型应用人才的重要环节。BOPPPS教学模式强调学生在课堂中的主动角色,秉持“以学为中心”的教学理念,围绕“参与式互动学习”的核心,引导学生进行探究式学习。本文结合微生物学与免疫学实验课程定位与目标,分析传统实验教学模式和BOPPPS教学模式的特点,以综合性实验“咽部细菌的革兰染色与药敏试验”为例,探索BOPPPS模式在实验教学中的具体应用模式,以期为提高微生物学与免疫学及相关课程的实验教学质量提供参考。

新医科背景下体内药物分析实验课程体系构建

体内药物分析实验课程是内蒙古医科大学药学类专业的专业课程,在“四新”建设背景下,高校要改变传统实验教学模式,积极开展新技术、新方法实验技能教学。文章通过分析体内药物分析实验课程教学存在的问题,积极开展体内药物分析实验课程体系的构建,以数字化教育为契机,开发虚拟仿真实验、优化线下实验内容、完善实验教学方法和实验考核评价方法,提高体内药物分析实验课教学水平,为建设体内药物分析实验一流课程奠定基础,同时为药学类其他专业课课程体系的建设提供参考。

高效科研方法融入药物分析实验教学

设计了采用高效液相色谱法测定药物含量的本科生化学实验。首先,配制对照品溶液和样品溶液,然后使用高效液相仪器进样。本实验可以锻炼学生制备不同梯度浓度样品的能力,熟悉供试品和对照品溶液的制备方法,掌握药物含量的测定方法,学会仪器的使用,并能了解实验数据分析,使学生进一步了解专业领域的前沿内容,提升自身科学素养和实践能力。

海洋产物作为制剂载体材料的研究进展

以“海洋多糖,海洋多肽,海洋多酚,药物制剂,载体,药物递送”中英文为关键词在中国知网数据库(CNKI)、百度学术、PubMed、Elsevier Science Direct等数据库进行检索,并从已获取文献的引文网络、核心文献推荐进行深度检索,通过阅读和分析最终纳入56篇文献进行分析。得出结论:海洋产物以其良好的生物相容性、可降解性、低毒、抗氧化、抗炎等特点,在生物医药材料领域受到广泛研究,大多用于靶向和缓释给药系统。本文从海洋产物来源、制备、应用进行综述。

前沿科研成果融入应用化学专业综合实验教学——新型二氧化硅交联胶束核壳纳米粒子在药物缓释方面的应用

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