温阳解毒化瘀颗粒对肝衰竭IETM大鼠TLR4 mRNA,TNF-α及肝细胞凋亡的影响

目的:研究Toll样受体4(TLR4) mRNA及其下游炎性因子肿瘤坏死因子-α(TNF-α)与肝衰竭肠源性内毒素血症(IETM)大鼠肝细胞凋亡的关系,并探索温阳解毒化瘀颗粒对内毒素介导的肝细胞凋亡的调控机制。方法:SPF级雄性SD大鼠85只,随机分为正常组,模型组,TLR4单克隆抗体组和温阳解毒化瘀颗粒组(8.925 g·kg^-1)。采用D-半乳糖胺(D-Gal)腹腔注射建立肝衰竭IETM模型,TLR4单克隆抗体组和温阳解毒化瘀颗粒组在造模前5 d给予温阳解毒化瘀颗粒溶液灌胃,正常组、模型组以等容积蒸馏水代替,直至处死。各组分别在24,48,72 h随机处死大鼠并采集标本。检测24,48,72 h各组时间点血清丙氨酸氨基转移酶(ALT),天门冬氨酸氨基转移酶(AST)水平,苏木素-伊红(HE)染色观察肝组织病理变化,实时荧光定量聚合酶链式反应(Real-time PCR)检测肝组织TLR4 mRNA表达,酶联免疫吸附测定法(ELISA)检测肝组织TNF-α表达,流式细胞仪检测肝细胞凋亡率。结果:与正常组比较,模型组ALT,AST升高,肝组织病理损伤程度明显加重,TLR4mRNA,TNF-α表达均增高(P<0.05,P<0.01),肝细胞凋亡率明显上升(P<0.05,P<0.01);与模型组比较,温阳解毒化瘀颗粒组ALT,AST显著降低(P<0.01),肝组织病理损伤程度明显减轻(P<0.05),TLR4 mRNA,TNF-α表达显著下降(P<0.01),肝细胞凋亡率亦显著降低(P<0.01)。结论:TLR4 mRNA,TNF-α在肝衰竭时与肝细胞凋亡呈正相关,温阳解毒化瘀颗粒能够改善肝衰竭IETM大鼠肝功能,减轻肝细胞损伤,减少肝细胞凋亡,其机制可能与其下调肝脏TLR4 mRNA表达,抑制TNF-α释放,降低肝细胞凋亡率有关。     

Redox active or thiol reactive? Optimization of rapid screens to identify less evident nuisance compounds

Compounds that exhibit assay interference or undesirable mechanisms of bioactivity are routinely encountered in assays at various stages of drug discovery. We observed that assays for the investigation of thiol-reactive and redox-active compounds have not been collected in a comprehensive review. Here, we review these assays and subject them to experimental optimization to improve their reliability. We demonstrate the usefulness of our assay cascade by assaying a library of bioactive compounds, chemical probes, and a set of approved drugs. These high-throughput assays should complement the array of wet-lab and in silico assays during the initial stages of hit discovery campaigns to pursue only hit compounds with tractable mechanisms of action.     

糖尿病小鼠小肠绒毛杯状细胞分泌黏原颗粒的变化

目的：观察糖尿病小鼠小肠绒毛杯状细胞分泌的黏原颗粒是否改变,为进一步研究糖尿病的消化系统并发症提供形态学基础。方法分别选取3,5,8,10月龄db/db糖尿病小鼠及相应年龄段的db/+m正常小鼠,每组6只,标本用4%多聚甲醛灌流固定后,从距Treitz韧带约10 cm处切下一段空肠,用PAS染色方法对小肠杯状细胞分泌的黏原颗粒进行观察。结果糖尿病组小肠绒毛易被破坏,糖尿病组中杯状细胞的分泌物比同月份正常对照组显著增加(P<0.05)；糖尿病组中3月龄的杯状细胞分泌物最多,8月龄最少,并且8月龄分别与3月龄、5月龄比较均存在着显著差异(P<0.05)；正常对照组随月龄增长无显著性差异(P>0.05)。结论糖尿病时杯状细胞分泌物增多可能对小肠绒毛有保护作用。     

基于“证治代谢组学”假说探讨中药（单体/复方）药效物质基础的研究思路与实践＊

中药(单体/复方)药效物质基础研究是中医药现代研究的关键问题之一。然而,目前运用的生化和分子生物学方法很难诠释其多靶点、整体性和动态性的整合调节作用。课题组提出的"证治代谢组学"假说理论指出,不同的证候存在"证相关代谢谱群"和"证相关生物标志物",这可能是中药药效物质基础所在,辨证论治后偏离的代谢网络功能呈现回归趋势。基于"证治代谢组学"假说开展中药(单体/复方)药效物质基础研究,这将为阐释中药(单体/复方)防病治病和养生保健的科学内涵及其对疾病个体的整合调节作用提供新的思路。本文以冠心病为研究载体、气阴虚血瘀证为切入点、活血保心丸为干预措施,就该研究思路的内涵与总体模式、提出的背景与依据、实践的方式与可行性、创新与特色及其研究意义等进行了简要介绍。     

不同组织膜蛋白及二甲基亚砜对小脑颗粒细胞突起生长的影响

目的探讨不同组织膜蛋白提取物和二甲基亚砜对大鼠小脑颗粒细胞（CGC）突起生长的影响。方法提取成年大鼠肝、坐骨神经和脑白质的组织膜蛋白包被盖片，接种CGC于盖片上，同时将二甲基亚砜（DMSO）作为添加剂加入培养液，观察CGC突起的生长。结果①脑白质膜蛋白能明显抑制CGC突起的生长，随浓度增加抑制效应更加明显；坐骨神经膜蛋白对CGC突起生长抑制不明显，肝组织膜蛋白则能促进突起生长；②随DMSO浓度升高CGC突起生长逐渐受到抑制，当DMSO浓度小于1％时，CGC突起长度与对照对照差别不大。结论脑白质膜蛋白对CGC突起生长有明显抑制作用，而DMSO添加到培养液中在低浓度时不会显著影响神经元突起生长，可用于检测药物及疏水性分子对体外培养的神经细胞突起生长的作用。     

复方碘酊消毒液中和剂试验研究

复方碘酊消毒液是含有20 g/L碘和1 g/L三氯羟基二苯醚的酒精溶液,为选择出合适的中和剂,采用化学中和法的悬液定量试验和滤膜冲洗及中和剂加滤膜冲洗法进行了试验研究。结果,硫代硫酸钠、硫代乙醇酸钠、吐温80、卵磷脂及其复方中和剂经悬液定量试验法和滤膜冲洗法,均不能中和复方碘酊消毒液对试验菌的残留作用。采用中和剂加滤膜冲洗法证明,用含5000 mg/L硫代硫酸钠+5000 mg/L吐温80+2000 mg/L硫代乙醇酸钠的磷酸盐缓冲液作为中和剂,均可有效去除复方碘酊消毒液对试验菌的残留作用;各中和剂及其中和产物对试验菌及培养基无不良影响。结论,选择复方碘酊消毒液的中和剂,不适宜用悬液定量试验法;宜采用滤膜过滤冲洗法。     

The effect of MS-818, a pyrimidine compound,on the regeneration of peripheral nerve fibers of mice after a crush injury

One of the pyrimidine compounds, 2-piperadino-6-methyl-5-oxo-5,6-dihydro(7H)pyrrolo[3,4-d]pyrimidine (MS-818), has neurotropic effects in vitro. Therefore, we studied the effect of MS-818 on the regeneration of the peroneal nerve in C57BL/6J mice after a crush injury. Two test groups, which received a daily intraperitoneal injection of 5 mg/kg or 10 mg/kg MS-818, respectively, were compared with controls, which received daily intraperitoneal injections of physiological saline, over a 14-day period. The maximum foot-width ratio (crushed side/uncrushed side) was obtained on days 1, 8 and 14 after the crush injury, and the various morphometric parameters were evaluated at both 5 and 10 mm distal to the proximal portion of the crush site. The significant effects of MS-818 included a larger maximum foot width (P<0.04) and a greater number of unmyelinated axons per nerve at both levels (P<0.003) in both test groups than in controls. MS-818 had no significant effects on body weight, the increase of total transverse fascicular area after the crush injury, the total number of myelinated fibers with their size distributions, or the number of nuclei of Schwann cells and macrophages. Therefore, we conclude that MS-818 promotes axonal sprouting and elongation after a crush injury in mice.     

肝积颗粒合介入治疗原发性肝癌的临床观察

目的：观察肝积颗粒合介入治疗原发性肝癌气虚血瘀证的临床疗效。方法：将符合研究条件的患者随机分为治疗组（30例）和对照组（30例），分别采用肝积颗粒保介入治疗和单纯介入治疗，观察两组的近期临床疗效、远期生存时间以及T细胞亚群、AFP值的变化情况。结果：治疗组瘤体缓解率为20．00％、稳定率为90．00％，对照组分别为16．67％、63．33％，其中，瘤体稳定率两组差异有显著性（P＜0．05），治疗组在减轻临床症状，延长生存时间、升高CD3、CD4值等方面优于对照组（P＜0．05），两组AFP值变化比较，差异无显著性。结论：肝积颗粒联合介入治疗原发性肝癌气虚血瘀证，可提高临床疗效，改善临床症状，增强机体免疫功能，延长生存期。