健脾中药诱导人胃癌细胞裸小鼠移植瘤细胞凋亡的实验研究

目的研究健脾类中药复方四君子汤及以四君子汤等健脾药物为基础结合清热解毒、软坚散结中药组成的复方SRRS对人胃癌细胞裸小鼠移植瘤的抑瘤作用与诱导肿瘤细胞凋亡的关系.方法采用人胃癌细胞SGC-7901裸小鼠皮下移植瘤模型.实验动物共30只,造模后随机分为3组,每组10只:SRRS方剂组,予240 g/L SRRS煎剂0.5mL/d灌胃;四君子汤组,予160 g/L四君子汤煎剂0.5 mL/d灌胃;对照组,予生理盐水0.5 mL/d灌胃,共40 d.应用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法(TUNEL法)、电镜,结合流式细胞仪分析技术,观察SRRS方剂及四君子汤的抑瘤作用与肿瘤细胞凋亡率及形态学改变.结果四君子汤、SRRS方剂对人胃癌细胞裸小鼠移植瘤抑瘤率分别为34%,47%,TUNEL法检测肿瘤细胞凋亡指数(AI)分别为:16%±3%,13%±3%,与对照组AI:3%±1%比较,显著提高(四君子组P＜0.01;SRRS方剂组P＜0.05,均为t检验);流式细胞分析示四君子汤组凋亡率:11%±6%,SRRS方剂组:12%±6%,均较对照组:7%±1%提高(t检验,P＜0.05);但四君子汤和SRRS方剂组之间的凋亡率TUNEL法和流式细胞分析均无差异.电镜下二中药治疗组见较多凋亡细胞和凋亡小体;对照组凋亡细胞和凋亡小体极少.结论中药四君子汤、SRRS方剂在体内的抑瘤作用与诱导胃癌细胞凋亡有关,其诱导细胞凋亡作用可能主要与健脾药物有泄?诱导肿瘤细胞凋亡可能是健脾药物抗癌作用的机制之一.     

介入治疗对兔VX2肝癌细胞增殖和凋亡的影响

目的研究介入治疗对肿瘤细胞凋亡和增殖的影响.方法建立兔VX2肝癌的动物模型,将实验动物分为介入组和对照组,用TUNEL法观察肿瘤细胞凋亡情况,采用原位杂交和免疫组化法检测凋亡和增殖相关基因bcl-2、bax及PCNA的表达情况.结果(1)介入组凋亡指数明显高于对照组,两组的细胞凋亡率分别为62.6 ±32.21%和21.4±10.43%(P＜0.05);介入组PCNA为38.80±15.73%,对照组为68.54±24.43%,前者显著低于后者(P＜0.05);(2)介入组bcl-2表达率为10.34±6.66%,显著低于对照组的24.5±11.53%(P＜0.05),而介入组bax的表达率为58.27±38.33%,显著高于对照组的32.13±23.76%(P＜0.05).bcl-2和bax基因mRNA表达与蛋白的表达相一致.结论抑制肿瘤细胞增殖和诱导其凋亡是介入治疗肝癌的重要机制之一.     

健脾理气方药物血清对肝癌细胞端粒酶活性及凋亡的影响

目的 观察健脾理气方药物血清对肝癌细胞SMMC-7721端粒酶活性的影响以及诱导肿瘤细胞凋亡的作用,探讨其抗肿瘤治疗的作用机制.方法 采用端粒重复序列扩增(TRAP)结合非变性聚丙烯酰胺凝胶电泳银染法,利用健脾理气方中药灌服新西兰兔后制备含药血清,观察健脾理气方药物血清对体外培养的人肝癌细胞株SMMC-7721端粒酶活性的影响,同时利用流式细胞仪检测和荧光显微镜观察对细胞凋亡的影响.结果 健脾理气药物血清在d 4开始对肿瘤细胞的端粒酶活性有抑制作用,电泳条带光密度明显降低;并能诱导肿瘤细胞的凋亡,凋亡的比例随作用时间的延长而升高,流式细胞术检测d2,d4,d 6的凋亡比例分别为0.81%,5.16%,8.45%.结论 对肿瘤细胞端粒酶活性的抑制和诱导肿瘤细胞凋亡是健脾理气中药抗肿瘤治疗作用机制的一部分.     

苦参素注射液联合顺铂对人肝癌SMMC-7721细胞凋亡相关基因c-myc、bcl-2和bax表达的影响

目的探讨苦参素注射液(MI)联合顺铂(DDP)对人肝癌SMMC-7721细胞凋亡相关基因c-myc、bcl-2和bax表达的影响。方法分别采用MI、顺铂及MI联合顺铂干预SMMC-7721细胞。TUNEL法检测细胞凋亡率,半定量RT-PCR法检测c-myc、bcl-2和bax mRNA表达,二步法免疫组化检测c-myc、bcl-2和bax蛋白表达。结果苦参素组、顺铂组和联合用药组细胞凋亡率显著上升,与对照组(不干预)比较差异有统计学意义(P<0.05或P<0.01),联合用药具有单纯相加或协同作用;联合用药组的细胞凋亡率显著增加,bax mRNA和蛋白表达显著升高,c-myc、bcl-2 mRNA和蛋白表达显著减少,与DDP单药组比较差异有统计学意义(P<0.05或P<0.01)。结论苦参素注射液联合顺铂具有单纯相加或协同诱导肝癌SMMC-7721细胞凋亡作用,其机制可能与bax基因表达上调和c-myc、bcl-2基因表达下调有关。     

牛磺酸对兔动脉粥样硬化血管平滑肌细胞凋亡的影响

目的:探讨牛磺酸对兔动脉粥样硬化血管平滑肌细胞凋亡及凋亡相关基因bcl-2、bax和caspase-3蛋白表达的影响.方法:将24只日本大耳白兔随机分为正常对照组、高脂模型组和牛磺酸组,14周后观察各组主动脉组织病理形态学改变,透射电镜观察斑块内平滑肌细胞超微结构变化,流式细胞术检测动脉粥样斑块内平滑肌细胞凋亡率,免疫组化染色法检测凋亡相关基因bcl-2和bax蛋白表达,Western blot蛋白印记法检测caspase-3蛋白表达.结果:高脂模型组与正常对照组比较,动脉内膜出现典型的斑块,管腔极度狭窄,典型的凋亡平滑肌细胞明显增多,平滑肌细胞凋亡率、bax及caspase-3蛋白表达升高(P<0.01),bcl-2蛋白表达降低(P<0.05).牛磺酸组与高脂模型组比较,主动脉斑块缩小,动脉管腔狭窄减轻,凋亡平滑肌细胞不典型且较少,平滑肌细胞凋亡率、bax蛋白表达及caspase-3蛋白表达降低(P<0.01),bcl-2蛋白表达升高(P<0.05).结论:牛磺酸可能通过对bcl-2、bax及caspase-3蛋白表达的调控而降低动脉斑块内过度凋亡的平滑肌细胞,从而抑制粥样斑块的形成.     

塞来昔布联合氟伐他汀对人肝癌裸鼠皮下移植瘤生长及细胞凋亡的影响

目的 探讨塞来昔布联合氟伐他汀对实验性人肝癌裸鼠皮下移植瘤生长及细胞凋亡的影响.方法 32只实验裸鼠左腋窝皮下接种BEL-7402肝癌细胞株,随机分为对照组、塞来昔布组、氟伐他汀组及塞来昔布和氟伐他汀联合用药组.实验结束时,留取移植瘤标本,流式细胞术及原位缺口末端标记法检测肿瘤细胞凋亡率,Western blot检测Akt、磷酸化Akt(p-Akt)和survivin蛋白的表达情况.数据比较采用析因设计多因素方差分析及多个样本均数间多重比较的SNK-q检验.结果 联合用药组肿瘤生长明显被抑制,塞来昔布组、氟伐他汀组抑瘤率分别为34.0%和25.0%,联合用药组抑瘤率为72.2%.对照组细胞凋亡指数为3.5%±0.8%,联合用药组为19.4%±3.0%,塞来昔布组和氟伐他汀组分别为8.5%±1.4%和9.4%±1.7%,联合用药组肿瘤细胞凋亡明显高于塞来昔布及氟伐他汀单药组(P值均＜0.01).流式细胞术检测结果显示,移植瘤细胞凋亡率对照组为4.1%±1.6%,塞来昔布组为9.1%±2.1%,氟伐他汀组为10.1%±2.3%,联合用药组为23.6%±5.8%,各单药用药组均高于对照组(P值均＜0.05),其中联合用药组高于对照组及各单药组(P值均＜0.01).Western blot检测结果显示,联合用药组较对照组明显下调p-Akt(0.23±0.08比1.12±0.07)和survivin蛋白(0.50±0.07比1.47±0.19)的表达(P值均＜0.01).结论 与单用塞来昔布或氟伐他汀相比,联合用药能更有效地抑制肝癌细胞生长.     

羟基磷灰石纳米粒子诱导肝癌SMMC-7721细胞凋亡后bcl-2和bax表达的变化

目的探讨凋亡相关基因bcl-2和bax在羟基磷灰石（HAP）纳米粒子诱导肝癌SMMC-7721细胞凋亡后表达的变化。方法采用免疫组化法检测bcl-2蛋白和bax蛋白表达。结果不同浓度的HAP纳米粒子处理细胞24、48、72h后bcl-2蛋白的表达降低，bax蛋白的表达升高，与对照组比较均有显著性差异，并且均呈现时间和剂量依赖性。结论。HAP纳米粒子诱导肝癌SMMC-7721细胞凋亡，与凋亡调控基因bax表达增强、bcl-2表达减少有关。     

钯配合物对人乳腺癌细胞MCF-7增殖的抑制作用及机制

目的 探讨钯配合物(PBIPS)对人乳腺癌细胞(MCF-7)增殖的抑制作用及其机制.方法 MTT法检测PBIPS对MCF-7的细胞毒性,荧光显微镜观察细胞形态学变化,流式细胞术(FCM)检测MCF-7细胞的凋亡率;Western-blotting检测不同浓度PBIPS作用48 h凋亡蛋白survivin 、caspase-3及bcl-2和bax的表达情况.结果 PBIPS可以时间和剂量依赖性地抑制MCF-7细胞的生长;与对照组比较,PBIPS不同浓度剂量组AO/EB荧光染色出现典型的凋亡形态学改变;FCM检测表明MCF-7细胞凋亡率各实验组均明显高于对照组(P＜0.01),且呈时间剂量依赖性;Western blotting显示与对照组相比,各实验组PBIPS可显著下调MCF-7细胞survivin和bcl-2的表达(P＜0.01),而caspase-3和bax的表达显著增加(P＜0.01).结论 PBIPS能明显诱导人乳腺癌MCF-7细胞凋亡,其机制可能与改变凋亡相关基因survivin、bcl-2、caspase-3及bax的表达有关.     

cDNA微阵列技术检测胃肠安复方对人胃癌细胞基因表达谱的影响

目的:观察健脾类中药胃肠安复方对人胃癌细胞裸小鼠移植瘤的抑瘤作用,探讨基因表达谱芯片在中药复方对胃癌细胞多基因表达影响研究中的应用.方法:采用人胃癌细胞SGC-7901裸小鼠皮下移植瘤模型.造模1周后选瘤块直径1.5 mm以上者共16只,随机分为胃肠安复方组及0.85%氯化钠溶液对照组,每组8只.造模后41 d处死动物.通过RT-PCR反转录、同位素标记、全基因表达谱芯片方法研究胃肠安复方组与对照组人胃癌移植瘤细胞mRNA的表达差异.结果:胃肠安复方组瘤重低于对照组[(0.99±0.49)g:(1.93±0.52) g,P<0.05],胃肠安复方组抑瘤率为48.64%.全基因表达谱芯片检测比较分析示,胃肠安复方组与对照组样品间共有45个显著性差异表达基因,其中上调表达24个,下调表达21个,35个已克隆基因,10个表达序列标签.结论:健脾中药胃肠安复方抑制人胃癌细胞裸小鼠移植瘤生长,并可能对肿瘤细胞多基因表达有影响.     

健脾理气药诱导人肝癌细胞SMMC7721凋亡的研究

目的观察健脾理气药的诱导凋亡效应,为其临床应用进一步提供依据.方法采用血清药理学方法研究中药的体外效应.应用四甲基偶氮唑蓝(MTT)比色法检测含中药兔血清对肝癌细胞的抑制效应,以Annexin V标记法、DNA含量测定、电子显微镜方法检测含中药血清诱导凋亡及细胞周期阻滞效应.免疫组化法观察含中药血清对P53,P21WAF1/CIP1蛋白的影响,RT-PCR法观察含中药血清对P21WAF1/CIP1 mRNA表达水平的影响.结果含中药血清有一定的抑制肝癌细胞作用,其作用3 d的抑制率为6.6%,作用6 d的抑制率为36.2%.含中药血清作用2 d诱导9.8%±4.0%的肝癌细胞凋亡,使细胞周期阻滞于S期,并上调P53蛋白、P21WAF1/CIP1 mRNA及蛋白的表达.结论健脾理气药具一定的诱导凋亡及抑制肝癌细胞效应,上调p53,p21WAF1/CIP1基因的表达为分子机制.     

胰岛素瘤的解剖分布特点分析

目的胰岛素瘤是最常见的胰腺神经内分泌肿瘤，因其临床表现多样，导致诊断困难。影像学诊断尤其是超声内镜(EUS)在胰岛素瘤的诊断中起着重要作用，拥有较高的敏感性和特异性。本研究拟通过明确胰岛素瘤的解剖分布特点，以期有助于提高影像学的诊断准确率和降低漏诊率，尤其是在教育和培训实践中对于EUS的学习者更具有指导价值。 方法回顾性分析解放军总医院第一医学中心病案资料数据库1993年1月至2019年11月经外科手术、病理确诊为胰岛素瘤的患者的临床资料，检索方法采取搜索术后病理诊断为"胰岛素瘤"的病例，通过查阅病例的方法，提取出胰岛素瘤的大小和解剖分布等数据，进一步分析其特点。 结果共检索到确诊为胰岛素瘤的患者116例，其中，男45例、女71例，年龄13～76岁，平均年龄(44.4±14.85)岁。胰岛素瘤单发110例(94.8%)、多发6例(5.2%)。位置分布：头颈部46例(39.7%)，单发45例、多发1例；体尾部68例(58.6%)，单发65例、多发3例；全胰腺多发2例(1.7%)。病变大小特点：最大径0.4～3.4 cm，平均大小(1.53±0.58)cm。≤1 cm 29例、>1 cm而≤1.5 cm41例、>1.5 cm而≤2.0 cm28例，≤3 cm 15例，>3 cm 3例。年龄与肿瘤的大小相关，≤44岁患者肿瘤平均大小为(1.36±0.51)cm、>44岁患者肿瘤平均大小为(1.70±0.60)cm，P<0.05。头颈部的肿瘤大于体尾部的肿瘤，头颈部肿瘤平均大小(1.66±0.63)cm，体尾部(1.42±0.52)cm，P<0.05。 结论胰岛素瘤在胰腺体尾部较头颈部更好发；绝大多数单发，但可以全胰腺多发；多数小于1.5 cm，肿瘤的大小与患者年龄和肿瘤的解剖分布相关。     

Pathogenesis of carcinoma of the papilla of Vater

Most adenomas and carcinomas of the small intestine and extrahepatic bile ducts arise in the region of the papilla of Vater. In familial adenomatous polyposis (FAP) it is the main location for carcinomas after proctocolectomy. In many cases symptoms due to stenosis lead to diagnosis at an early tumor stage. In about 80%, curative intended resection is possible. Operability is the most relevant prognostic factor. Most ampullary carcinomas resp. carcinomas of the papilla of Vater develop from adenomatous or flat dysplastic precursor lesions. They can be sited in the ampulloduodenal part of the papilla of Vater, which is lined by intestinal mucosa. They also can develop in deeper parts of the ampulla, which are lined by pancreaticobiliary duct mucosa. Intestinal-type adenocarcinoma and pancreaticobiliary-type adenocarcinoma represent the main histological types of ampullary carcinoma. Furthermore, there exist unusual types and undifferentiated carcinomas. Many carcinomas of intestinal type express the immunohistochemical marker profile of intestinal mucosa (keratin 7?, keratin 20+, MUC2+). Carcinomas of pancreaticobiliary type usually show the immunohistochemical profile of pancreaticobiliary duct mucosa (keratin 7+, keratin 20?, MUC2?). Even poorly differentiated carcinomas, as well as unusual histological types, may conserve the marker profile of the mucosa they developed from. These findings underline the concept of histogenetically different carcinomas of the papilla of Vater which develop either from intestinal- or from pancreaticobiliary-type mucosa of the papilla of Vater. Molecular alterations in ampullary carcinomas are similar to those of colorectal as well as pancreatic carcinomas, although they appear at different frequencies. In future studies, molecular alterations in ampullary carcinomas should be correlated closely with the different histologic tumor types. Consequently, the histologic classification should reflect the histogenesis of ampullary tumors from the two different types of papillary mucosa.     

Lack of correlation of degree of villous atrophy with severity of clinical presentation of coeliac disease

BACKGROUND AND AIM: Both the clinical presentation and the degree of mucosal damage in coeliac disease vary greatly. In view of conflicting information as to whether the mode of presentation correlates with the degree of villous atrophy, we reviewed a large cohort of patients with coeliac disease. PATIENTS AND METHODS: We correlated mode of presentation (classical, diarrhoea predominant or atypical/silent) with histology of duodenal biopsies and examined their trends over time. RESULTS: The cohort consisted of 499 adults, mean age 44.1 years, 68% females. The majority had silent coeliac disease (56%) and total villous atrophy (65%). There was no correlation of mode of presentation with the degree of villous atrophy (p=0.25). Sixty-eight percent of females and 58% of males had a severe villous atrophy (p=0.052). There was a significant trend over time for a greater proportion of patients presenting as atypical/silent coeliac disease and having partial villous atrophy, though the majority still had total villous atrophy. CONCLUSIONS: Among our patients the degree of villous atrophy in duodenal biopsies did not correlate with the mode of presentation, indicating that factors other than the degree of villous atrophy must account for diarrhoea in coeliac disease.     

Demonstration of the mechanism of action of biguanides

Summary Palmitic acid oxidation in rat diaphragm homogenate is depressed by biguanide concentrations that are still incapable of inhibiting oxidative phosphorylation. Glucose oxidation is not directly effected by the same biguanide concentrations: however, the inhibitory effect of palmitic acid on glucose oxidation is partly removed by biguanides. Inhibition of fatty acid oxidation, which accounts for most of the metabolic effects caused by these drugs, can be regarded as the fundamental mechanism of action of biguanides. There is some evidence suggesting that these drugs might interact with carnitine, thus preventing long-chain fatty acids from being transported across the mitochondrial membrane to the site of oxidation. Traduzione a cura degli AA.     

血吸虫童虫生长发育及其机制的研究进展

血吸虫童虫是宿主免疫系统攻击的重要靶标,包括皮肤型、肺型和肝门型童虫。宿主分子对童虫生长发育具有重要作用。童虫生长发育机制包括免疫调节、信号转导、性别发育及凋亡等。肌动蛋白、组织蛋白酶、烯醇化酶和葡萄糖基转移酶等分子为血吸虫童虫生长发育的重要分子。本文对血吸虫童虫生长发育及其机制的研究进展做一综述。     

124例耐多药结核分枝杆菌基因突变特征分析

目的对临床分离的耐多药结核分枝杆菌相关基因的突变特征进行分析。方法对124例耐多药结核分枝杆菌以及50株敏感株的耐药相关基因(包括异烟肼inh A、kat G、oxyR-ahp C间隔区以及利福平rpo B)进行序列测定,分析其基因突变情况。结果异烟肼耐药inh A基因突变率为14.5%;kat G基因突变率为70.2%(87/124),主要位于315位;oxyR-ahp C间隔区突变率为15.3%;inh A、kat G两种基因同时突变率75.0%,三种基因同时突变率为89.5%。利福平rpo B基因突变的检出率高达95.2%,突变主要发生在531、526、516位点。结论我省耐多药菌异烟肼耐药相关基因最常见突变为kat G 315、inh A C-T(-15)、axyR-ahp C间隔区(-10)C-T,利福平为rpo B531、526、516。结合MDR-TB耐药相关基因的特征分析,可以建立一种快速、准确、特异的适合于我省的检测结核菌耐多药性的新方法。     

Assessment of quality of life of parents of children with juvenile chronic arthritis

The aim of the study was to assess the quality of life (QOL) and the psychological status of parents of children with juvenile chronic arthritis (JCA). The QOL, anxiety and depression of the parents of 28 children with JCA were evaluated and compared to those of the parents of 28 healthy children. Mothers of JCA children and mothers of healthy children reported similar QOL. The reported anxiety and depression levels were similar for mothers and fathers in both groups. The parents of children with pauciarticular-type JCA reported lower QOL and higher levels of anxiety and depression than the parents of children with other types, namely polyarticular and systemic JCA. These findings may be explained by the fact that the pauciarticular patients had shorter disease duration and were less frequently seen in the outpatient clinic. The QOL of mothers of children with JCA was found to be slightly impaired in the group of children with pauciarticular JCA. Future larger studies are needed to confirm these results, as the number of subjects in the three groups was rather low. Received: 26 September 2001 / Accepted: 8 February 2002     

Numerical Simulation of the Effect of Rate of Change of Glucose on Measurement Error of Continuous Glucose Monitors

Background

A 5-day in-patient study designed to assess the accuracy of the FreeStyle Navigator® Continuous Glucose Monitoring System revealed that the level of accuracy of the continuous sensor measurements was dependent on the rate of glucose change. When the absolute rate of change was less than 1 mg•dl⁻¹•min⁻¹ (75% of the time), the median absolute relative difference (ARD) was 8.5%, with 85% of all points falling within the A zone of the Clarke error grid. When the absolute rate of change was greater than 2 mg•dl⁻¹•min⁻¹ (8% of the time), the median ARD was 17.5%, with 59% of all points falling within the Clarke A zone.

Method

Numerical simulations were performed to investigate effects of the rate of change of glucose on sensor measurement error. This approach enabled physiologically relevant distributions of glucose values to be reordered to explore the effect of different glucose rate-of-change distributions on apparent sensor accuracy.

Results

The physiological lag between blood and interstitial fluid glucose levels is sufficient to account for the observed difference in sensor accuracy between periods of stable glucose and periods of rapidly changing glucose.

Conclusions

The role of physiological lag on the apparent decrease in sensor accuracy at high glucose rates of change has implications for clinical study design, regulatory review of continuous glucose sensors, and development of performance standards for this new technology. This work demonstrates the difficulty in comparing accuracy measures between different clinical studies and highlights the need for studies to include both relevant glucose distributions and relevant glucose rate-of-change distributions.     

Angiographic diagnosis of the degree of serosal invasion of carcinoma of the colon

Angiography using Prostaglandin El^® was performed on 38 patients with carcinoma of the colon in order to diagnose the degree of serosal cancer invasion. The findings at angiography were classified into four groups:1) AG-S3, abnormal change (irregularity and/or encasement) up to marginal vessels; 2) AG-S2, abnormality up to vasa recta; 3) AG-S1, abnormality of penetrating branches of vasa recta within the wall of the colon; and 4) AG-S0, no distinct findings of abovementioned vessels. These angiographic findings were compared with both macroscopic and microscopic serosal cancer invasion. Angiographic diagnosis is in accord with the macroscopic findings in 84.2 percent of cases. Angiographic diagnosis is in accord with the microscopic findings in 32.4 percent of cases. Macroscopic findings confirm the angiographic diagnosis precisely but the conflict with microscopic findings should not be overlooked. This may be the result of inflammatory change, adhesion, and fibrosis around carcinoma of the colon.     

Study of constancy of hydrogen-consuming flora of human colon

The constancy of the hydrogen consuming flora of the human colon was studied in 15 healthy subjects via two measurements obtained 18 to 36 months apart. Hydrogen disappearance rate and the major products of H₂-consuming bacteria, methane and sulfide, were measured during incubation of fecal homogenates with excess hydrogen and sulfate. In 11/15, the hydrogen consumption rate and the predominant hydrogen-consuming pathway (methanogenesis, sulfate reduction, or neither) remained constant. However, major shifts in these pathways were observed in four subjects, with two losing and two gaining the ability to produce methane. Methanogenesis was associated with the highest hydrogen consumption rate. This study demonstrates that clinically unrecognizable, major alterations of the colonic flora occur in healthy subjects. Understanding of the factors responsible for these alterations might allow for therapeutic manipulation of the colonic flora.Supported in part by the Department of Veterans Affairs and NIDDKD RO1 DK 13309-25.