膈下逐瘀汤加减方对人肝癌细胞端粒酶的影响

目的：探讨膈下逐瘀汤加减方（JGXZY）对人肝癌细胞端粒酶活性的影响。方法：制备5组试验血清：不含药物血清（W组）,阳性对照药物复方斑蝥胶囊含药血清（B组）和JGXZY低（5%）、中（10%）、高（15%）浓度含药血清（分别为D组、Z组、G组）。观察各组药物血清对端粒酶活性的影响。结果：JGXZY各血清组与B组在抵制人肝癌细胞端粒酶活性方面比较差异有显著性意义（P〈0.05或P〈0.01）,其中G组血清的作用最强。结论：GXZY对人肝癌细胞端粒酶活性有明显的抑制作用。     

膈下逐瘀汤加减方对肝癌H22荷瘤小鼠的作用

[目的]研究膈下逐瘀汤加减方对肝癌H22荷瘤小鼠的作用。[方法]以肝癌H22荷瘤小鼠为模型,观察膈下逐瘀汤加减方的抗肿瘤作用,并分别计算抑瘤率和观察瘤体组织学形态及检测甲胎蛋白(AFP)、γ-谷氨酰氨转肽酶(γ-GT)值。[结果]相当于生药0.4、0.8、1.2 g/kg的膈下逐瘀汤加减方对肝癌H22荷瘤小鼠的抑瘤率均在48%以上,与模型组比较差异有统计学意义(P<0.01),与复方斑蝥胶囊(阳性对照)组比较差异无统计学意义(P>0.05);瘤体组织形态学变化及瘤体AFP、γ-GT值与模型组比较差异均有统计学意义(均P<0.01)。[结论]膈下逐瘀汤加减方对肝癌H22荷瘤有明显抑制作用。     

参苓白术散和膈下逐瘀汤对实验性脂肪肝早期的影响

目的：比较早期应用活血化瘀法和健脾益气法两种治疗法则对高脂饲料导致的大鼠脂肪肝模型的影响。方法：饲喂高脂饲料复制大鼠脂肪肝模型，造模的同时分别以健脾益气法代表方参苓白术散和活血化瘀法代表方膈下逐瘀汤进行干预,观察两方对模型大鼠早期的血脂、肝脂、肝功能以及超氧化物歧化酶（SOD）、丙二醛（MDA）、血小板聚集的影响。结果：两方均能降低模型大鼠的血脂、肝脂水平，从作用强度来看，膈下逐瘀汤作用优于参苓白术散,膈下逐瘀汤能够改善肝功能，参苓白术散对肝功能作用不明显；两方均能升高SOD，作用以膈下逐瘀汤更为显著,对MDA两方均无明显影响；参苓白术散对模型大鼠血小板聚集没有明显作用，而膈下逐瘀汤早期应用能够显著对抗血小板聚集。结论：早期应用活血化瘀法对改善脂肪肝预后有重要意义。     

中药复方搏癌丸对人肝癌细胞周期及bax、bcl-2、p53基因表达调控的影响

目的:研究中药复方搏癌丸对人肝癌细胞HepG2周期及相关调控基因表达的影响.方法:采用药物血清和细胞药理学方法:①用流式细胞仪观察搏癌丸对人肝癌HepG2细胞凋亡及周期的影响.②用免疫组织化学染色(SABC)法检测搏癌丸3个有效药物血清浓度对bax、bcl-2、p53基因表达的调控.结果:①10%搏癌丸药物血清作用HepG2细胞48小时后,FCA检测的细胞凋亡率为25.34%,均显著高于相应无药血清对照组的3.54%,P＜0.05.②能上调HeG2细胞凋亡调控基因bax、p53的表达,与无药血清对照组比较,差异有显著性意义,P＜0.05,且呈剂量依赖关系.结论:搏癌丸诱导HepG2细胞凋亡的作用机制之一可能是上调促凋亡基因bax和p53的表达.     

热处理对人肝癌细胞HepG2基因表达谱的影响

肿瘤的热疗已成为近年来肿瘤治疗学的一个热点[1] 。为了从基因水平上深入研究热杀灭细胞机制 ,本文应用基因点数为 12 5 5 0的人类基因表达谱芯片 ,筛查经热处理的人肝癌细胞ＨｅｐＧ2 出现表达变化的基因 ,探索这些基因的变化与热杀伤肿瘤细胞的内在联系与规律 ,为肿瘤的热疗提供分子生物学依据。一、材料与方法1.主要试剂及设备 :Ｔｒｉｚｏｌ试剂盒、Ｔ7 (Ｔ) 2 4引物、ＳｕｐｅｒＳｃｒｉｐｔⅡＲｅｖｅｒｓｅＴｒａｎｓｃｒｉｐｔａｓｅ、ＤＮＡｌｉｇａｓｅ、ＤＮＡ多聚酶Ⅰ、ＲＮＡ酶Ｈ(Ｇｉｂｃｏ) ;ＭＥＧＡｓｃｒｉｐｔＴ7ＩｎＶ…     

皂荚提取物对人肝癌细胞相关癌基因表达的调控作用及端粒酶的影响

目的：研究皂荚提取物对人肝癌细胞的增殖、相关癌基因表达的调控作用及端粒酶的影响。方法：体外培养人肝癌细胞bel-7402，检测不同浓度皂荚提取物（0．05mg／ml、0．1mg/ml、0．2mg／m1）对人肝癌细胞bel-7402细胞增殖、细胞周期、肝癌相关基因（bax、bcl-2、p53）的表达及端粒酶活性的影响。结果：0．05mg／ml-0．2mg／ml浓度组皂荚提取液对人肝癌细胞bel-7402细胞增殖、调控癌基因与抑癌基因、促进癌细胞凋亡、抑制端粒酶活性差异均具有显著性意义（P〈0．05或P〈0．01）。结论：皂荚提取物能够抑制人肝癌细胞增殖，促进细胞凋亡。     

健脾理气方对HepG2人肝癌细胞株p53和bcl-2基因表达的影响

[目的]观察健脾理气方对HepG2人肝癌细胞株p53和bcl-2基因表达的影响。[方法]30只雄性SD大鼠随机分为健脾理气方（中药）组、0．85％氯化钠（对照）组和顺铂（化疗）组,每组10只。利用血清药理学方法制备药物血清,3组分别加入PRMI1640血清培养的HepG2人肝癌细胞株中,作用48h。应用免疫细胞化学方法和图像分析方法观察健脾理气方对HepG2人肝癌细胞株p53、bcl-2基因表达影响。[结果]①免疫细胞化学法显示抑癌基因p53主要定位在细胞核,癌基因bcl-2主要定位在细胞质内。药物血清作用后,p53表达明显增强;bcl-2表达明显降低。随着作用时间延长,2种基因表达改变更明显。②中药组与对照组p53和bcl-2光度（AOD）值差异有统计学意义（P〈0．01）。③化疗组作用结果与中药组相似。[结论]健脾理气方药物血清上调p53、下调bcl-2,达到抑制HepG2人肝癌细胞株的作用。为应用健脾理气方治疗肝癌提供实验依据。     

芍药甙诱导人肝癌细胞系HepG2凋亡的研究

目的：观察芍药甙诱导人肝癌细胞系HepG2凋亡及凋亡相关基因的变化。方法：用0．5～0．2g／L芍药甙处理人肝癌HepG2细胞，光镜下观察细胞形态；MTT法检测细胞增殖率；流式细胞仪检测凋亡峰；免疫组化法检测P53，bcl-2，bax等凋亡相关基因的表达。结果：0．5～2．0g／L浓度的芍药甙对HepG2有显著抑制作用；可诱导HepG2凋亡，并能上调凋亡相关基因P53和bax表达、下调抗凋亡基因bcl-2表达。结论：0．5～2．0g／L芍药甙可诱导人肝癌细胞HepG2凋亡，其机制可能与凋亡相关基因表达增强和抗凋亡相关基因表达减弱有关。     

蟾毒灵对人肝癌BEL-7402细胞增殖的影响

目的：研究蟾毒灵对人肝癌BEL-7402细胞增殖及细胞周期的影响。方法：采用MTT法观察蟾毒灵对人肝癌细胞增殖的抑制作用；光镜及透射电镜观察细胞形态及超微结构；流式细胞术分析细胞周期分布情况。结果：蟾毒灵对肝癌细胞生长抑制作用呈浓度-时间依赖性，24、48和72小时的半数抑制浓度分别为6．67、0．348和0．228μmol／L，蟾毒灵可将细胞周期阻滞于G2／M期，透射电镜观察到典型的细胞凋亡特征。结论：诱导肝癌细胞周期G2／M期阻滞可能是蟾毒灵抑制肝癌细胞增殖的机制之一。     

膈下逐瘀汤治疗酒精性肝病的作用机制探讨

目的 基于网络药理学、分子对接的方法分析膈下逐瘀汤治疗酒精性肝病的作用机制,并对其进行动物模型实验验证。方法 基于TCMSP数据库筛选膈下逐瘀汤潜在活性化合物和作用靶点,并通过GeneCards、OMIM、PharmGkb、TTD和DrugBank数据库检索酒精性肝病疾病相关靶点,二者取交集,获得膈下逐瘀汤治疗酒精性肝病的关键靶点。对关键靶点进行功能富集分析和分子对接。通过STRING平台和Cytoscape软件构建膈下逐瘀汤治疗酒精性肝病的潜在活性化合物—靶点调控网络和蛋白互作网络,并对网络进行拓扑分析,得到最终核心网络和靶点。取SPF级雄性SD大鼠60只,随机取12只作为正常对照组,其余48只以乙醇灌胃的方法制备酒精性肝病模型,将其分为模型对照组、水飞蓟素组、膈下逐瘀汤低剂量和高剂量组各12只。膈下逐瘀汤低剂量组和高剂量组分别给予6.2、12.4 g/kg膈下逐瘀汤水煎液灌胃,水飞蓟素组予以水飞蓟素0.2g/kg灌胃,模型对照组和正常对照组予以同等体积蒸馏水灌胃,连续灌胃6周。取大鼠腹主动脉血检测血清丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）、甘油三酯（TG）、γ谷氨...     

Correlation between expression of gastrin, somatostatin and cell apoptosis regulation gene bcl-2／bax in large intestine carcinoma

AIM: To explore the correlation between expression of somatostatin (SS), gastrin (GAS) and cell apoptosis regulation gene bcl-2/bax in large intestine carcinoma. METHODS: Sixty-two large intestine cancer tissue samples were randomly and retrospectively selected from patients with large intestine carcinoma. Immunohistochemical staining for bcl-2, bax, GAS, SS was performed according to the standard streptavidin-biotin-peroxidase (S-P) method. According to the semi-quantitative integral evaluation, SS and GAS were divided into three groups as follows. Scores 1-3 were defined as the low expression group, 4-8 as the intermediate expression group, 9-16 as the high expression group. Bax and bcl-2 protein expressions in different GAS and SS expression groups of large intestine carcinoma were assessed. RESULTS: The positive expression rate of bax had a prominent difference between SS and GAS high, intermediate and low expression groups (P<0.05, x2SS = 9.246; P<0.05, x2GAs = 6.981). The positive expression rate of bax in SS high (80.0%, 8/10) and intermediate (76.5%, 13/17) expression groups was higher than that in low expression group (40.0%, 14/35) (P<0.05, X2high vslow = 5.242; P<0.05, x2middle vs low = 6.097). The positive expression rate of bax in GAS high expression group (27.3%, 3/8) was lower than that in low expression group (69.4%, 25/36) (P<0.05, x2 = 4.594). However, bax expression in GAS intermediate expression group (46.7%, 7/15) was lower than that in low expression group, but not statistically significant. The positive expression rate of bcl-2 had a prominent difference between SS and GAS high, intermediate and low expression groups (P<0.05, x2ss = 7.178; P<0.05, x2GAS = 13.831). The positive expression rate of bcl-2 in GAS high (90.9%, 10/11) and intermediate (86.7%, 13/15) expression groups was higher than that in low expression group (44.4%, 16/36) (P<0.05, x2high,vslow = 5.600; P<0.05, x2middle vs low = 7.695). However, the positive expression rate of bcl-2 in SS high (40. 0%, 4/10) and intermediate (47.1%, 8/9) expression groups was lower than that in low expression group (77.1%, 27/35) (P<0.05, x2high vs low = 4.710; P<0.05, x2middle vs low = 4.706). There was a significant positive correlation between the integral ratio of GAS to SS and the integral of bcl-2 (P<0.01, r = 0.340). However, there was a negative correlation between the integral ratio of GAS to the SS and bax the integral of (P<0.05, r= -0.299). CONCLUSION: The regulation and control of gastrin, somatostatin in cell apoptosis of large intestine carcinoma may be directly related to the abnormal expression of bcl-2, bax.     

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。     

胰腺癌Bcl-2,P53蛋白表达和细胞凋亡

目的　探讨ｂｃｌ２ ,ｐ５３ 基因和细胞凋亡在胰腺癌发病机制中的作用以及它们之间相互关系．方法　应用ＡＢＣ 免疫组化技术检测５０ 例胰腺癌中Ｂｃｌ２ 和Ｐ５３ 蛋白表达,运用原位末端标记法观察肿瘤中细胞凋亡数量．结果　Ｐ５３ 蛋白表达阳性率为５４ ％ ,临床Ⅰ期阳性率（２６－７ ％ ）却显著低于Ⅱ期（６１－１ ％ ） 和Ⅲ＋ Ⅳ期（７０－６ ％ ,Ｐ＜ ０－０５） ;Ｂｃｌ２蛋白表达阳性率为６４ ％ ,临床Ⅰ期阳性率（９３－３ ％ ） ,显著高于Ⅱ期（５５－６ ％ ） 和Ⅲ＋ Ⅳ期（４７－１ ％ ,Ｐ＜ ０－０５） ;组织学Ⅲ级癌细胞中凋亡指数明显高于Ⅰ,Ⅱ级（ Ｐ＜ ０－０５） ,Ｂｃｌ２ 蛋白阴性病例中凋亡指数明显高于Ｂｃｌ２ 阳性者（ Ｐ＜ ０－０１） ．结论　Ｂｃｌ２ 是通过抑制细胞凋亡参与肿瘤的生长过程,Ｂｃｌ２和Ｐ５３ 蛋白表达之间 存在密切负相关（τ＝ － ０－１７４７ ,Ｐ＜ ０－０５） ．     

Prognostic significance of bcl-2, bax,and p53 expression in diffuse large B-cell lymphoma

Diffuse large B-cell lymphoma (DLCL) exhibits heterogeneous clinical features and varies markedly in response to treatment and prognosis. Because apoptosis-related proteins may play an important role in predicting the prognosis of DLCL, the current study investigated the prognostic significance of a high level of bcl-2, bax, and p53 expression in relation to clinical characteristics in patients with DLCL. Paraffin-embedded specimens from 94 patients with de novo DLCL were analyzed immunohistochemically for bcl-2, bax, and p53 gene expression. Cases with a positive immunohistological stain in more than 50% of the tumor cells were considered to have DLCL-positive expression. Patients were treated optimally, i.e., with radiotherapy including brief cycles of CHOP or CHOP-like regimens for patients with stage 1-2A diseases and with at least 6 cycles of CHOP or CHOP-like regimens for stage 2B-4 diseases. The responses to therapy and survival were then analyzed in 94 uniformly staged patients. bcl-2 expression was identified in 24 patients (26.4%), bax expression in 35 patients (37.6 %), and p53 expression in 21 patients (22.6%). bax expression proved to be a statistically significant prognostic factor in predicting the overall survival (OS) (P = 0.0015) and disease-free survival (DFS) (P = 0.0052), regardless of other clinical factors or immunohistological results. There was no significant difference in the OS (P = 0.0682) or DFS (P = 0.088) between the bcl-2-positive (n = 24) and bcl-2-negative (n = 67) groups. However, bcl-2 expression was found to be unfavorably associated with the OS (P = 0.0054) in a confined group with low (n = 51) or low intermediate (n = 22) IPI scores. The expression of p53 exhibited no statistical correlation with the OS or DFS. A multivariate analysis revealed that IPI score, bulky mass, and bax expression were all significantly associated with the DFS or OS. bax and bcl-2 should be considered as independent biologic prognostic parameters in DLCL, thereby aiding in the identification of patient risk groups. As such, bcl-2-positive patients with a low or low intermediate IPI score, or without a high level of bax expression could be candidates for more intensive therapy or alternative therapeutic approaches.     

凋亡抑制基因survivin bcl-2 bax在肺癌中表达的研究

目的检测肺癌患者癌组织和癌旁组织中survivin、bcl-2及bax的基因表达,探讨它们的相关性及与肺癌发生、发展的关系。方法应用TUNEL原位细胞凋亡检测方法及免疫组化方法,对1998—2004年华中科技大学同济医学院附属协和医院收治的163例肺癌患者手术常规石蜡包埋组织中癌基因survivin、bcl-2及bax的表达进行检测,并与其中106例患者的癌旁组织对比,分析免疫组化结果及其与肺癌的病理特征和预后关系。结果在癌旁肺组织中,survivin、bcl-2、bax蛋白阳性表达率分别为1·9%(2例)、29·2%(31例)、93·47%(99例);肺癌组织内,三者阳性表达率分别为69·9%(114例)、62·0%(101例)、52·8%(86例)。异常增高的survivin、bcl-2表达呈明显相关。结论细胞凋亡和增殖失控,相关基因survivin、bcl-2和bax蛋白异常表达在肺癌发生发展中起重要作用。survivin、bcl-2可能在肺癌癌变及浸润过程中起着重要作用,可作为判断肺癌生物学行为和预后的参考指标。     

萎胃汤治疗慢性萎缩性胃炎伴癌前病变临床疗效及对PCNA和bal-2、bax表达的影响

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