胃痞颗粒对胃癌癌前病变患者胃黏膜上皮细胞凋亡及Bcl-2 P53 Fas蛋白表达的影响

目的观察胃痞颗粒对胃癌癌前病变患者黏膜上皮细胞凋亡及调控蛋白表达的影响。探讨胃痞颗粒治疗胃癌癌前病变的作用机制。方法46例患者随机分为治疗组30例,对照组16例,治疗组用胃痞颗粒一号与胃复春片一号,对照组用胃痞颗粒二号与胃复春片二号,连续应用6个月,进行相关检测。结果治疗组患者细胞凋亡指数治疗后比治疗前明显提高,也比对照组治疗后升高显著;Bcl-2、P53、Fas蛋白表达结果治疗组与对照组相比差异均有统计学意义。结论胃痞颗粒对胃黏膜癌前病变有逆转治疗作用。胃痞颗粒通过抑制Bcl-2、P53(突变型)蛋白表达,促进Fas蛋白表达和细胞凋亡指数增加,从而有利于胃黏膜异型增生的上皮细胞发生凋亡,这可能是胃痞颗粒治疗癌前病变获效的机制之一。     

胃和冲剂Ⅱ号对慢性萎缩性胃炎大鼠胃黏膜三叶因子1和CD34表达的影响

目的 观察胃和冲剂Ⅱ号对慢性萎缩性胃炎(CAG)大鼠胃黏膜三叶因子1(TFF1)和CD34表达的影响,探讨胃和冲剂Ⅱ号治疗CAG的作用机制.方法 将72只雄性Wistar大鼠随机分为正常对照组(12只)和造模组(60只).除正常对照组外,其余各组大鼠均使用综合造模法成功复制CAG动物模型(为确定造模成功,颈椎脱臼法处死9只大鼠).然后将53只CAG大鼠随机分为5组,分别为模型组(10只)、维酶素阳性对照组(11只)及胃和冲剂低、中、高剂量组(10只,11只,11只).治疗60d后,取各大鼠相同部位全层胃壁组织,采用免疫组化法分别检测胃黏膜组织的TFF1和CD34的表达水平.结果 TFF1在胃和冲剂高剂量组中的表达值(3.4±1.4),高于模型组(1.5±0.8)和维酶素组(1.7±0.9)(P＜0.01);胃和冲剂高剂量组微血管密度值为(7.5±1.5),低于模型组(12.5±2.5)和维酶素阳性对照组(11.6±2.1),差异有统计学意义(P＜0.01).结论 胃和冲剂Ⅱ号可能通过提高胃黏膜组织的TFF1表达、降低CD34的表达,加快胃黏膜细胞的修复和抑制过度增殖,从而对CAG起到治疗作用并可预防癌变.     

地塞米松通过影响Caspase-3及NF-κB表达加重脑缺血再灌注凋亡作用

目的研究地塞米松是否加重脑缺血再灌注细胞凋亡及核转录因子NF-κB、Caspase-3在此过程中的作用.方法采用线栓法建立大鼠局灶性脑缺血再灌注模型.缺血1 h后生理盐水组腹腔注射0.9%氯化钠注射液(0.5mL),地塞米松组腹腔注射地塞米松[0.5 mg/(kg·d)].利用TUNEL法研究凋亡细胞的变化,应用免疫组化、原位杂交法检测NF-κB蛋白、Caspase-3 mRNA的表达.结果各缺血组凋亡细胞显著增多.地塞米松组各时点凋亡细胞数多于生理盐水组,NF-κB活化程度均明显低于生理盐水组,Caspase-3染色阳性细胞数明显高于生理盐水组(P＜0.01).结论地塞米松加重大鼠局灶性脑缺血再灌注损伤可能是与其阻碍NF-κB蛋白活化与上调Caspase-3 mRNA表达,并继而加重脑神经细胞凋亡的机制有关.     

消痞灵冲剂对大鼠实验性萎缩性胃炎伴不典型增生胃黏膜CEA及rasP21变化的影响

目的：从胃黏膜癌胚抗原(CEA)及rasP21变化的角度进一步探讨实验性慢性萎缩性胃炎伴不典型增生发病机理及消痞灵冲剂对其防治的有效机制。方法：采用综合法(幽门弹簧插入加口服盐水及热糊法)复制大鼠慢性萎缩性胃炎伴不典型增生模型，免疫组化法显示胃黏膜组织CEA及rasP21表达。结果：模型组及自然恢复组大鼠胃黏膜CEA及rasP21阳性表达明显增强，消痞灵预防及治疗组CEA表达则明显减弱(P＜0．05)，rasP21表达有一定程度减弱(P＞0．05)。结论：抑制胃黏膜CEA及rasP21阳性表达是消痞灵冲剂防治大鼠萎缩性胃炎伴不典型增生有效机制之一，其效果明显优于阳性对照药维酶素片。     

健脾清化中药复方对大鼠慢性萎缩性胃炎TLR4-MyD88依赖途径蛋白表达及TNF-α的影响

目的观察健脾清化中药复方对慢性萎缩性胃炎(CAG)大鼠TLR4及下游My D88依赖途径相关蛋白表达及炎性因子TNF-α的影响,探讨健脾清化中药复方治疗CAG的分子机制。方法将53只Wistar大鼠随机分为空白组8只和CAG造模组45只,以"氨水+去氧胆酸钠+乙醇"法复制CAG大鼠模型。确认造模成功后,将造模组余下40只CAG大鼠随机分为模型组、维酶素组、中药低、中、高剂量组,各8只。各组给予相应药物灌胃,连续30 d。HE染色观察病理组织学改变,Western blot法检测TLR4、My D88、NF-κB、COX-2的蛋白表达量,ELISA法检测血清中TNF-α含量。结果模型组大鼠TLR4、My D88、NF-κB、COX-2蛋白表达水平明显增高(P<0.01),血清TNF-α含量明显增高(P<0.01)。与模型组比较,健脾清化中药复方低、中、高剂量组胃黏膜病变明显改善,TLR4、My D88、NF-κB、COX-2蛋白表达水平均明显下降(P<0.05或P<0.01),血清TNF-α含量降低(P<0.05或P<0.01)。结论健脾清化中药复方可有效改善CAG大鼠胃黏膜组织病理改变,其治疗机制可能与降低组织中TLR4-My D88依赖途径中相关蛋白表达,以及抑制炎症因子的表达有关。     

复方蒲黄颗粒对局灶性脑缺血再灌注大鼠神经元凋亡及Caspase-3 mRNA表达的影响

目的:观察复方蒲黄颗粒(PH)对局灶性脑缺血再灌注大鼠神经元凋亡的影响,并探讨其作用机制。方法:采用大脑中动脉线栓法建立局灶性脑缺血再灌注大鼠模型。75只大鼠分为假手术、模型和PH高、中、低剂量组。采用Nissl染色观察神经元形态变化,TUNEL法检测神经元凋亡,原位杂交检测Caspase-3mRNA的表达。结果:PH高、中剂量组少量细胞核固缩深染,细胞丢失、凋亡细胞数及Caspase-3mRNA阳性细胞数均显著减少。结论:PH可下调Caspase-3的表达,从而抑制神经元凋亡,对脑缺血再灌注损伤有保护作用。     

红景天苷对局灶性缺血-再灌注损伤后神经细胞凋亡及对Bax、Bcl-2的影响

目的 研究红景天苷对局灶性脑缺血再灌注大鼠大脑皮层神经细胞凋亡和Bax、Bcl-2表达的影响.方法 将SD雄性大鼠随机分为假手术组,模型对照组,尼莫地平组,红景天苷低、中、高剂量组.采用线栓法建立大鼠大脑中动脉缺血再灌注损伤模型,并对各组给予相应的实验药物,然后在缺血再灌注24h和72h后处死所需动物,采用TTC染色法检测脑梗死体积,TUNEL标记法和免疫组化法分别检测观察细胞凋亡数目和Bcl-2、Bax表达的阳性细胞数.结果 与模型组相比,红景天苷各剂量组均可缩小大鼠脑皮质的梗死体积(P＜0.05或P＜0.01),降低残存细胞中TUNEL阳性细胞数(P＜0.05或P＜0.01);增加Bd-2表达阳性细胞数,降低Bax表达的阳性细胞教,升高Bcl-2/Bax的表达比例(P＜0.01).总体上24 mg·kg-1剂量组作用较好,优于阳性对照药尼莫地平(P＜0.05或P＜0.01).结论 红景天苷有明显的抗凋亡作用,其机制可能与增加Bcl-2阳性细胞的表达,降低Bax阳性细胞的表达,提高Bcl-2/Bax表达比有关.     

黄芪联合红花对脑出血大鼠出血灶周围神经细胞凋亡的影响

目的 比较早期不同时间联合使用黄芪和红花治疗脑出血（ICH）模型大鼠对出血灶周围神经细胞凋亡的影响.方法 ICH大鼠模型制造成功后,在出血前24h和出血后0、6、12、24h分别用红花、黄芪、黄芪＋红花治疗,72h后处死,作TUNEL染色和Caspase-3免疫组织化学染色.结果 出血对照组血肿周边及外围Caspase-3阳性细胞明显较多.黄芪组、红花组及黄芪＋红花组脑出血大鼠血肿周围Caspase-3阳性细胞计数均少于出血对照组,并且黄芪＋红花组的Caspase-3阳性细胞数明显低于红花组（P〈0.01）.黄芪＋红花术前24h组、术后0、6、12h脑出血大鼠的血肿周围Caspase-3阳性细胞数差异无统计学意义（P〉0.05）.但黄芪＋红花组术后应用24h脑血肿周围Caspase-3阳性细胞数均多于术前应用24h,术后0、6h和术后12h,差异有统计学意义（P〈0.05或P＜0.01）.黄芪组、红花组和黄芪＋红花组术后6、24h大鼠脑血肿周围TUNEL阳性细胞数均少于对照组,且红花组TUNEL阳性细胞数多于黄芪＋红花组,差异均有统计学意义（P＜0.01）.结论 联合使用黄芪和红花治疗脑出血优于单纯使用红花治疗脑出血;ICH 24h以后联合黄芪和红花治疗脑出血的效果较ICH以前24h给药治疗好;联合使用黄芪和红花对脑出血可能的保护机制是,通过抑制细胞内的Caspase-3蛋白的表达而延缓细胞凋亡.     

活血清下法在小肠缺血再灌注时对细胞凋亡因子Apaf-1和Bcl-2的调控作用

目的观察活血清下法对小肠缺血再灌注动物黏膜细胞凋亡蛋白酶活化因子-1(Apaf-1)和B淋巴细胞瘤-2(Bcl-2)的调控作用,并探讨其对肠屏障的保护机制.方法选用健康Wistar大鼠,随机分为对照组、模型组、活血承气合剂治疗组.对动物模型再灌注6、24、72 h时血浆内毒素、小肠黏膜上皮细胞凋亡因子Apaf-1和Bcl-2的表达分别进行检测和观察.结果模型组和活血承气组6、24 h的内毒素水平明显高于对照组(P＜0.01),而活血承气组各时间点内毒素水平均低于模型组(P＜0.01);活血承气组Apaf-1阳性细胞表达明显低于模型组(P＜0.01),并在72 h低于对照组(P＜0.05),模型组Apaf-1表达明显高于对照组(P＜0.01);活血承气组Bcl-2表达明显高于模型组(P＜0.01),在72 h高于对照组(P＜0.05),而模型组Bcl-2表达明显低于对照组(P＜0.05).结论小肠缺血再灌注早期肠黏膜上皮细胞凋亡是造成内毒素明显升高、肠屏障功能损伤的主要原因之一.活血清下法可以通过抑制促凋亡因子Apaf-1表达及促进抑凋亡因子Bcl-2的表达来减弱肠黏膜上皮细胞的凋亡,从而达到降低内毒素水平,保护肠屏障功能的作用.     

米诺环素对大鼠局灶性脑缺血再灌注后半胱天冬酶12表达的影响

目的:探讨米诺环素对大鼠局灶性脑缺血再灌注后缺血半暗带半胱天冬酶12(Caspase-12)表达的影响.方法:用大脑中动脉栓塞法建立大脑局灶性缺血再灌注模型.36只雄性Wistar大鼠,随机分为假手术组(Sham组),缺血再灌注组(IR组)和米诺环素干预组(Minocycline组),每组12只.采用DNA原位末端缺口标记法(TUNEL)检测神经元凋亡,免疫组织化学法检测组织Caspase-12蛋白的表达,反转录-聚合酶反应法(RT-PCR)检测Caspase-12mRNA表达水平.结果:与Sham组相比,IR组缺血半暗带凋亡神经元数增加(P＜0.05),Caspase-12表达增加(P＜0.05);米诺环素处理后显著缓解凋亡细胞数的增加,抑制Caspase-12的表达(P＜0.05),但仍高于Sham组(P＜0.05).结论:抑制Caspase-12介导的Caspase介导的级联反应凋亡途径的激活是米诺环素缺血再灌注损伤细胞凋亡的机制之一.     

Role of glutathione in reduction of arsenate and of gamma-glutamyltranspeptidase in disposition of arsenite in rats

Arsenate (AsV), the environmentally prevalent form of arsenic, is converted sequentially in the body to arsenite (AsIII), monomethylarsonic acid (MMAsV), monomethylarsonous acid (MMAsIII), and dimethylarsinic acid (DMAsV) and some trimethylated metabolites. Although the biliary excretion of arsenic in rats is known to be glutathione (GSH)-dependent, involving transport of arsenic-GSH conjugates, the role of GSH in the reduction of AsV to the more toxic AsIII in vivo has not been defined. Therefore, we studied how the fate of AsV is influenced by buthionine sulfoximine (BSO), which depletes GSH in tissues. Control and BSO-treated rats were given AsV (50 micromol/kg, i.v.) and arsenic metabolites in bile, urine, blood and tissues were analysed by HPLC-HG-AFS. BSO increased retention of AsV in blood and tissues and decreased appearance of AsIII in blood, bile (by 96%) and urine (by 63%). The biliary excretion of MMAsIII was also nearly abolished, the appearance of MMAsIII and MMAsV in the blood was delayed and the renal concentrations of these monomethylated arsenicals were decreased by BSO. Interestingly, appearance of DMAsV in blood and urine remained unchanged and the concentrations of this metabolite in the kidneys and muscle were even increased in response to BSO. To test the role of gamma-glutamyltranspeptidase (GGT) in arsenic disposition, the effect of the of the GGT inhibitor acivicin was investigated in rats injected with AsIII (50 micromol/kg, i.v.). Acivicin lowered the hepatic and renal GGT activities and increased the biliary as well as urinary excretion of GSH, but failed to alter the disposition (i.e. blood and tissue concentrations, biliary and urinary excretion) of AsIII and its metabolites. In conclusion, shortage of GSH decreases not only the hepatobiliary transport of arsenic, but also reduction of AsV and the formation of monomethylated arsenic, while not hindering the production of dimethylated arsenic. While GSH plays an important role in the disposition and toxicity of arsenic, GGT, which hydrolyses GSH and GSH conjugates, apparently does not influence the fate of the GSH-reactive trivalent arsenicals in rats.     

微透析取样技术在中药体内分析中的应用研究进展

本文综述了微透析取样技术在中药体内分析中的应用,介绍微透析取样技术的原理、组成、探针类型、特点,重点阐述了微透析取样技术在测定脑、血液、皮肤等组织器官中中药有效成分浓度的应用实例。表明微透析取样技术在中药药效研究中具有广阔的前景。     

应用PASS系统对我院2008年住院医嘱的分析

目的监测分析2008年我院住院患者用药情况。方法将PASS系统嵌入医生工作站、临床药学工作站等子系统,构建合理用药计算机网络系统,对住院医嘱进行及时监测,将监测结果向医生反馈,并对其进行统计、分析。结果2008年共监测医嘱3 620 241条,不合理医嘱908条,占0.02%。不合理医嘱中,配伍禁忌(381条)占41.96%,用法用量(381条)占41.96%,药物相互作用(108条)占11.89%,儿童用药(38条)占4.19%。经与医生沟通后,更改不合理医嘱856条,占94.27%。结论PASS系统可有效监测医嘱中的不合理用药,通过与医生交流,大大减少药物不良事件的发生,值得临床推广应用,也为临床药师开展工作带来了极大的便利。但PASS系统尚存在局限性,有待进一步完善。     

Role of desacetylation in the detoxification of cephalothin in renal cells in culture

The toxicity of three cephalosporin antibiotics to rabbit kidney cells in culture was compared to their known nephrotoxic potential in vivo (cephaloridine greater than cefazolin greater than cephalothin). While cephalothin is considered to be a relatively nonnephrotoxic cephalosporin when administered to many species including humans and rabbits, in several in vitro systems involving rabbit renal tissue, cephalothin was comparatively more toxic than anticipated based on in vivo data. Cephalothin is extensively desacetylated in rabbits to a less microbiologically active metabolite, desacetylcephalothin. When a microsomal S9 fraction from rabbit kidney was added to the in vitro assay in cultured rabbit renal cells, cephalothin was desacetylated and its toxicity to kidney cells was reduced. The addition of S9 in vitro provided a toxicity ranking of the cephalosporins that correlated with their known in vivo nephrotoxic potentials (cephaloridine greater than cefazolin greater than cephalothin). The in vitro detoxification of cephalothin by S9 was blocked by the coadministration of the esterase inhibitor, aminocarb. Desacetylcephalothin was relatively nontoxic to rabbit renal tissue in vitro. These results suggest that the desacetylation of cephalothin in vivo represents a previously unrecognized mechanism of detoxification of this cephalosporin antibiotic. Furthermore, this mechanism of detoxification may be applicable to other acetylated cephalosporins.     

2009年某院住院患者抗真菌药用药调查

目的:分析讨论某院抗真菌药使用的合理性,为临床安全有效地使用抗真菌药提供参考。方法:回顾性统计分析某院2009年住院患者抗真菌药用药信息。结果:2009年某院住院患者抗真菌药DDDs排名前3名分别为:氟康唑、制霉菌素和伊曲康唑;使用金额排名前3名分别为:氟康唑、米卡芬净及卡泊芬净;更换一种抗真菌药进行治疗的患者数为176人,在全部患者中占13.4%。结论:应进一步强化用药指征的意识,提高标本送检率,同时改善某些抗真菌用药不合理更换的现象,以避免耐药性发生,从而更好更长远地体现抗真菌药的治疗价值。     

应用PASS系统分析我院2010年住院医嘱

目的:了解我院2010年住院患者的合理用药情况,探讨如何利用合理用药监测系统( PASS)提高合理用药水平.方法:利用PASS对我院2010年15 966例住院患者的1 184 997条用药医嘱进行监测,以黑色警示医嘱为依据,收集不合理用药信息,并对监测结果进行统计、分析.结果:不合理用药医嘱50 261条,发生率为4.24％.绝对禁止黑色医嘱5441条,主要为药物相互作用(66.54％)、注射液体外配伍(17.86％)、用法用量(15.46％)、儿童警告(1.14％).结论:应用PASS系统能有效监测医嘱中的不合理用药情况,有利于提高临床合理用药水平,但PASS系统尚存在局限性,有待进一步完善.     

Kinetics of in vitro metabolism of methoxyphenamine in rats

1. Methoxyphenamine (MP) was metabolized in vitro by rat liver preparations to O-desmethylmethoxyphenamine (O-desmethyl-MP), N-desmethylmethoxyphenamine (N-desmethyl-MP) and 5-hydroxymethoxyphenamine (5-hydroxy-MP). These metabolic pathways were inhibited by SKF 525-A and carbon monoxide, which indicates that these reactions were mediated at least partly by an NADPH-dependent cytochrome P-450 system. 2. Strain differences in the metabolism of this drug in vitro were observed in female Lewis and Dark Agouti (DA) rats, which are proposed models for human debrisoquine phenotypes. Methoxyphenamine O-demethylase and 5-hydroxylase activity in DA rats were lower than those in Lewis rats. 3. The metabolic transformation of methoxyphenamine in vitro to O-desmethyl-MP was inhibited competitively by debrisoquine and sparteine. This indicates that the cytochrome P-450 isoenzyme mediating the metabolism of MP to O-desmethyl-MP is similar to that mediating metabolism of debrisoquine and sparteine. However, no inhibition was observed with methenytoin.     

Changes in levels of dependency and predictors of mortality in elderly people in institutional care in Dunedin

The 1983 study of dependency of subjects in institutional care in Dunedin was repeated two years later. A significant increase in levels of dependency in residential homes, particularly in the Religious and Welfare sector was found. In 1983 there were 29 high dependency residents and 73 medium dependency residents in residential homes. In 1985 these numbers had increased to 55 and 86 respectively. There was no change in the number of low dependency residents. In 1983, 6 high dependency residents had been admitted to residential home care in the year prior to the study. In 1985 the number of high dependency residents recently admitted had increased to 23. There had also been a significant increase in the dependency of patients in Religious and Welfare continuing care hospitals. Of the 933 subjects in institutional care in 1983 who were able to be followed, 354 (37.9%) died in the following 2 years. Mortality rate was higher for those in hospital care (48.1%) than for those in residential home care (29.6%). Mortality rates were higher in more dependent subjects and this was evident for each measure of dependency.     

Study of potential in vitro and in vivo genotoxicity in hepatocytes of quinolone antibiotics

The genotoxicity of quinolone antibiotics has been evaluated in hepatocytes following in vitro and in vivo exposure. Unscheduled DNA synthesis (UDS) was induced in vitro in rat hepatocytes by norfloxacin, ofloxacin, pefloxacin, and ciprofloxacin but not by nalidixic acid. In vivo UDS was not observed in hepatocytes isolated 4 to 24 hr after exposure of adult male F344 rats to either a single dose (30 to 190 mg/kg) or repeated doses (40 mg/kg) of ciprofloxacin. Using the 32P-postlabeling technique, no modified bases were detected in hepatocytes exposed in vitro to ciprofloxacin. In summary, UDS was induced in hepatocytes by in vitro exposure to high concentrations of norfloxacin, ofloxacin, pefloxacin, or ciprofloxacin. There was no evidence of in vitro DNA adduct formation by ciprofloxacin or in vivo DNA damage under the conditions tested. These findings suggest that ciprofloxacin is not DNA reactive, but it induces in vitro UDS as a consequence of some indirect action.