截断法与逆挽法治疗慢加急性肝衰竭的网络药理学研究

目的:探讨截断逆挽方中截断法与逆挽法治疗慢加急性肝衰竭(ACLF)的生物学内涵,为深入研究该方的疗效机制提供支持。方法:借助TCMSP及BATMAN-TCM中药数据库,设置ADME参数筛选截断逆挽法的活性成分并收集对应靶点,构建活性成分-靶点相互作用网络及靶点蛋白质-蛋白质相互作用(PPI)网络,富集分析靶点参与的生物学过程和KEGG信号通路,分析方剂的潜在生物学机制。比较截断法和逆挽法在活性成分、靶点PPI网络及KEGG信号通路方面的异同。结果:截断法关联88个药物活性成分、逆挽法关联32个药物活性成分,其中有6个共有成分。IL6、VEGFA、TP53、TNF、MAPK1在截断法和逆挽法靶点PPI网络中均具有重要作用。截断法和逆挽法靶点共同参与抗ACLF信号通路,包括神经活性配体-受体相互作用、TNF信号通路、HIF-1信号通路、PI3K-Akt信号通路、乙型肝炎等。此外,截断法与P53信号通路相关,逆挽法与PPAR信号通路相关。结论:截断逆挽方在抗ACLF中具有多靶点、多途径、综合调控的特点,截断法和逆挽法可产生协同作用,截断法侧重抗凋亡,逆挽法与脂质代谢、氧化应激及免疫调控有关。     

阿尔茨海默病模型大鼠脑海马内MMP-9和TIMP-1 mRNA的表达

目的探讨基质金属蛋白酶(MMP)-9和基质金属蛋白酶组织抑制因子(TIMP)-1在阿尔茨海默病(AD)模型大鼠脑海马CA1区的mRNA表达水平。方法将30只雄性Wistar大鼠,随机分为A、B、C、D、E组,每组6只。A组注射5μl生理盐水,B、C、D、E组双侧海马注射凝胶态Aβ25~355μl(依次含有Aβ25~350.5,1.0,5.0,10.0μg),大鼠于14 d后处死。取各组大鼠新鲜血清采用酶联免疫吸附试验(ELISA)检测肿瘤坏死因子(TNF)-α和白细胞介素(IL)-1β的表达量;取新鲜脑海马组织提取mRNA,qRTPCR技术检测MMP-9和TIMP-1 mRNA表达量。结果血清TNF-α含量D、E组明显高于A、B、C组(P<0.01);血清IL-1β含量D、E组明显高于A、B、C组(P<0.01)。MMP-9 mRNA表达量D、E组明显高于A、B、C组(P<0.01);TIMP-1 mRNA表达D、E组明显高于A、B、C组(P<0.01)。结论 Aβ所致AD大鼠脑内发生了炎症反应,MMP-9表达上调,而TIMP-1可与MMP-9特异性结合,进而抑制其活性,因此随着MMP-9上调,TIMP-1表达量也随之增加。     

益生菌对慢加急性肝衰竭大鼠模型的保护作用及其机制

目的探究益生菌干预对慢加急性肝衰竭(ACLF)大鼠的作用及其机制。方法采用随机数字表法将44只雄性SD大鼠随机分为6组,对照组1(C1组,n=6,不做任何干预)、模型组1(M1组,n=8,40%CCl4油溶液腹腔注射10周,从第7周开始每天灌胃PBS溶液,10周末给予D-GalN急性攻击)、益生菌干预组1(Y1组,n=8,造模同M1组,灌胃剂为益生菌溶液)、对照组2(C2组,n=6,不做任何干预)、模型组2(M2组,n=8,腹腔注射40%CCl4油溶液,10周后给予D-GalN急性攻击,48 h后每天灌胃PBS溶液,持续到第12周末处死)、益生菌干预组2(Y2组,n=8,造模同M2组,灌胃剂为益生菌溶液)。观察干预前后大鼠体质量、肝功能、肝组织病理学,采用ELISA法测血浆内毒素、肠道分泌型免疫球蛋白A(s IgA),Western Blot法测定肠道紧密连接蛋白occludin的水平,RT-PCR法检测肠道紧密连接蛋白occludin和ZO-1的mRNA水平,通过选择性培养基测定肠道菌群等指标的变化。计量资料多组间比较采用单因素方差分析,进一步两两比较采用SNK-q检验。结果 C1、M1、Y1 3组间与C2、M2、Y2 3组间体质量、肝指数、ALT、AST、TBil、血浆内毒素、s IgA、occludin mRNA、ZO-1mRNA水平比较差异均有统计学意义(F1值分别为27. 65、8. 96、61. 37、18. 27、21. 00、87. 01、67. 10、101. 50、105. 40,P值均0. 05; F2值分别为14. 04、12. 85、14. 02、11. 39、35. 80、19. 14、15. 37、25. 02、126. 00,P值均0. 05),C1、M1、Y1 3组间occludin蛋白水平比较差异有统计学意义(F=16. 40,P 0. 05)。C1、M1、Y1 3组间与C2、M2、Y2 3组间乳酸杆菌、双歧杆菌、肠球菌、肠杆菌含量比较差异均有统计学意义(F_1分别为77. 95、66. 61、25. 63、33. 29,P值均0. 05; F_2分别为21. 50、22. 62、6. 71、17. 74,P值均0. 05)结论 ACLF大鼠出现肠道菌群紊乱和肠屏障功能障碍,益生菌干预可以重塑ACLF大鼠的肠道菌群结构,维护ACLF大鼠的肠屏障功能,促进ACLF大鼠肝脏的修复。     

结肠癌组织中细胞周期蛋白E和增殖细胞核抗原表达的研究

目的研究细胞周期蛋白E(CyclinE)和增殖细胞核抗原(PCNA)在结肠癌组织中的表达及二者之间的关系。方法采用原位杂交法和免疫组化S-P法,对40例结肠癌、10例结肠腺瘤和10例结肠正常黏膜组织进行CyclinE mRNA和PCNA的检测。结果结肠癌组织中CyclinEmRNA的阳性表达率为62.5%,PCNA指数为(50.09±11.15),均显著高于结肠腺瘤和结肠正常组织(P<0.05);CyclinE mRNA表达阳性组的PCNA指数明显高于CyclinE mRNA阴性组(P<0.05);CyclinE mRNA与肿瘤的浸润深度、有无淋巴结转移密切相关(P<0.05),与肿瘤的分化程度、有无肝转移不相关(P>0.05);PCNA与肿瘤的浸润深度、分化程度及有无淋巴结转移密切相关(P<0.05),与有无肝转移无明显相关(P>0.05)。结论CyclinE可作为结肠癌的又一细胞增殖指标,其对结肠癌的发生、发展起重要作用。     

慢性脑缺血大鼠海马区Cdc42表达及其与认知功能障碍的相关性

目的研究慢性脑缺血大鼠海马组织中细胞分裂周期蛋白42(Cdc42)表达的变化,探讨其在慢性脑缺血所致认知功能障碍中可能发挥的作用。方法大鼠40只,随机分为假手术组、持久性双侧颈总动脉结扎(2VO)8、10、12 w组,每组各10只。应用Morris水迷宫检测大鼠学习记忆能力,用免疫组织化学及Western印迹两种方法检测各组大鼠海马区Cdc42的表达。结果 Morris水迷宫显示手术组大鼠较假手术组大鼠逃避潜伏期明显延长(P<0.05);免疫组化及Western印迹均显示手术组大鼠海马区Cdc42表达明显低于假手术组(P<0.05)。结论 Cdc42表达量的降低可能参与了慢性脑缺血所致认知功能障碍的形成。     

针刺预处理对脑梗死大鼠胆碱能抗炎通路的影响研究

目的探讨针刺预处理对脑梗死大鼠胆碱能抗炎通路(CAP)的影响。方法选择40只健康成年SD雄性大鼠,采用随机数字表法将大鼠分为正常组(A组)、假手术组(B组)、脑梗死组(C组)、针刺预处理组(D组)和针刺预处理+甲基牛扁亭(MLA)组(E组),各8只。A组大鼠不做任何处理;B组大鼠只切开皮肤,暴露颈总动脉、颈外动脉、颈内动脉,不插入线栓,术后缝合皮肤;C组大鼠采用改良Zea Longa方法制备脑梗死模型;D组于造模前7 d开始针刺治疗;E组大鼠于针刺同时腹腔注射MLA,5 mg/kg,1次/d,连续治疗7 d。各组大鼠于造模成功后同步饲养,自由饮食,3 d后处死。于处死前采用Longar 5分制评分法进行神经功能评分;取处死大鼠新鲜脑组织顶叶皮质采用甲醛固定,采用TUNEL法检测细胞凋亡情况;剩余脑组织采用液氮保存,采用酶联免疫吸附试验(ELISA)检测TNF-α、IL-1β水平,采用实时定量荧光PCR检测α7烟碱型乙酰胆碱受体(α7n AChR)mRNA表达情况。结果 C组、D组、E组大鼠TUNEL阳性细胞多于A组和B组,脑组织中TNF-α、IL-1β水平高于A组和B组(P0.05);C组和E组大鼠α7n AChR mRNA相对表达量低于A组和B组(P0.05);D组大鼠TUNEL阳性细胞少于C组和E组,神经功能评分低于C组和E组,脑组织中TNF-α、IL-1β水平低于C组和E组,α7n AChR mRNA相对表达量高于C组和E组(P0.05)。结论针刺预处理可以改善脑梗死大鼠神经功能,抑制神经元凋亡,降低炎性细胞因子水平,上调α7n AChR mRNA的表达。     

部分肝切除后的肝损伤血清和肝细胞生长因子促进大鼠骨髓细胞表达甲胎蛋白和白蛋白

目的 检测大鼠骨髓中是否存在肝脏干细胞，并用肝损伤血清和肝细胞生长因子(HGF)刺激骨髓细胞向肝细胞转化。方法 SD大鼠骨髓单个核细胞分3组进行培养：(1)单纯培养基对照组；(2)肝损伤血清组(15％，血清来自于2-AAF 75％肝切除大鼠)；(3)HGF(20ng／ml)组。用甲胎蛋白(AFP)和白蛋白作为细胞标志，用免疫组织化学、逆转录聚合酶链反应(PCR)、巢式PCR和western blot方法，观察大鼠肝损伤血清和HGF对骨髓细胞转化的促进作用。结果 肝损伤血清组和HGF组培养后10d和20d AFP免疫组织化学和western blot染色阳性；RT-PCR AFP mRNA阳性，新鲜骨髓细胞和单纯IMDM／F12培养基组AFP蛋白和mRNA均阴性。新鲜骨髓细胞存在白蛋白mRNA表达，在肝损伤血清组和HGF组培养后10d和20d，白蛋白mRNA表达增强。结论 大鼠肝损伤血清和HGF可促使体外培养骨髓细胞表达AFP mRNA和AFP。骨髓中可能存在骨髓源性肝干细胞，并微弱表达白蛋白mRNA；肝损伤血清和HGF可以促进骨髓细胞表达白蛋白mRNA。     

肝组织解偶联蛋白2在梗阻性黄疸及胆道再通实验大鼠模型中的表达及意义

目的研究梗阻性黄疸及胆道再通大鼠肝组织解偶联蛋白(UCP)2的表达及意义,探讨梗阻性黄疸对能量代谢障碍与氧化损伤的影响。方法 36只健康雄性大鼠随机分为6组,每组6只:假手术组(A组)、梗阻性黄疸1周组(B组)、梗阻性黄疸2周组(C组)、梗阻性黄疸1周再通1周组(D组)、梗阻性黄疸10 d再通1周组(E组)、梗阻性黄疸2周再通1周组(F组)。检测各组动物血清ALT、DBil、TBil水平;光镜下观察肝脏形态学改变;采用逆转录-聚合酶链反应(RT-PCR)法检测大鼠肝脏UCP-2mRNA的表达。结果胆道梗阻后,B、C 2组血清TBil、DBil、ALT水平较A组均升高(P值均0.05),C组升高较B组明显(P值均0.05)。光镜下可见B、C组肝细胞肿胀,炎性细胞浸润,汇管区伴胆管增生,C组出现典型的肝细胞片状坏死。B、C 2组肝组织UCP-2 mRNA表达较A组升高(P值均0.05),C组升高较B组明显(P0.05)。胆道再通组后,D、E、F 3组血清TBil、DBil、ALT水平呈不同程度下降。光镜下观察肝脏形态改变也明显趋于正常。D组肝细胞UCP-2 mRNA的表达高于B组(P0.05),F组的表达低于C组(P0.05),D组的表达低于E、F 2组(P值均0.05),E、F 2组差异无统计学意义(P0.05)。结论梗阻性黄疸时肝脏UCP-2 mRNA表达升高,随梗阻时间延长其升高趋势明显。解除梗阻时间早,则肝细胞再生活跃,UCP-2 mRNA表达暂时升高;解除梗阻时间迟,则肝组织损伤严重,再生缓慢,UCP-2 mRNA表达进行性下降,但仍高于正常。提示肝组织UCP-2 mRNA表达升高可能是梗阻性黄疸能量代谢紊乱的主要原因之一。     

脉络宁对老年性痴呆模型大鼠脑组织SIRT1基因表达的影响

目的观察中药复方脉络宁和谢海洲方,西药维生素(C+E)和阿托伐他汀药物干预对老年痴呆(AD)模型大鼠脑组织SIRT1基因表达的影响。方法用去卵巢同时腹腔注射D-半乳糖大鼠模拟早期AD模型,予以中药复方脉络宁和谢海洲方,西药维生素(C+E)和阿托伐他汀干预,采用Morris水迷宫检测大鼠用药前后学习记忆能力的变化,采用RT-PCR,Western印迹技术检测大鼠颞叶右侧皮层SIRT1基因表达的变化。结果与假手术组相比,去卵巢并注射D-半乳糖组逃避潜伏期明显延长;给药后,与病理组相比,四个给药组总逃避潜伏期明显缩短。与假手术组相比,病理组的SIRT1 mRNA和蛋白质的相对表达量明显降低,而与病理组相比,四个给药组的SIRT1 mRNA和蛋白质的相对表达量均有不同程度的升高。结论本模型大鼠颞叶皮层SIRT1 mRNA和蛋白质表达水平均降低;而脉络宁,谢海洲方,维生素(C+E),阿托伐他汀均可以不同程度的升高SIRT1 mRNA和蛋白质的表达水平,提示SIRT1表达的增加可能是这些药物对AD大鼠模型的神经保护作用分子机制之一。     

蓝莓对免疫性肝纤维化大鼠细胞色素P4502E1表达的影响

目的 观察蓝莓对大鼠免疫性肝纤维化的预防作用及其对细胞色素P4502E1 (CYP2E1)表达的影响.方法 将50只健康清洁级Wistar大鼠随机分为正常对照组(A组)、肝纤维化模型组(B组)、蓝莓原浆预防组(C组)、复方鳖甲软肝片预防组(D组)、蓝莓原浆加复方鳖甲软肝片预防组(E组)共5组,每组10只.除正常对照组外,其余各组均腹腔注射猪血清制备大鼠肝纤维化模型,各预防组在造模同时分别给予蓝莓原浆或(和)复方鳖甲软肝片灌胃,1次/d,共12周.12周末处死大鼠,行肝脏病理组织学检查,测定各组大鼠血清ALT水平、肝组织匀浆超氧化物歧化酶(SOD)活性、丙二醛(MDA)及羟脯氨酸(Hyp)含量,采用实时荧光定量聚合酶联反应、Western blot和免疫组织化学法检测大鼠肝组织CYP2E1的mRNA及蛋白质表达. 结果 12周末处死大鼠,A、B、C、D、E各组大鼠血清ALT水平分别为(37.9±4.5) U/L、(49.2±9.8) U/L、(39.9±6.3) U/L、(40.5±5.7) U/L及(38.2±8.4)U/L,各组间差异无统计学意义;各预防组大鼠肝纤维化程度较B组明显减轻(F=95.097,P＜0.05); C、D、E各组大鼠肝组织匀浆Hyp分别为(472.7±44.1)μg/g、(416.1±39.4)μg/g和(429.5±55.1)μg/g,低于B组的(603.2±68.9) μg/g,F=39.315,P＜0.05,差异有统计学意义; SOD水平分别为(2.5±0.4) U/mg、(2.0±0.5)U/mg、(2.2±0.2) U/mg,高于B组的(1.6±0.4) U/mg,F=25.557,P＜0.05,差异有统计学意义;MDA分别为(0.83±0.06) mol/mg、(0.96±0.08) nmol/mg、(0.85±0.06)nmol/mg,低于B组的(1.24±0.15)nmol/mg,F=46.376,P＜0.05,差异有统计学意义;B、C、D、E组大鼠肝组织CYP2E1的mRNA及蛋白质表达高于A组,C、D、E组大鼠肝组织CYP2E1的mRNA及蛋白质表达低于B组,但差异均无统计学意义.结论 蓝莓对猪血清所致大鼠肝纤维化有一定的预防作用;免疫性肝纤维化时,大鼠肝组织CYP2E1的表达无明显改变;蓝莓对CYP2E1的表达无明显影响.     

慢加急性肝衰竭小鼠肝组织组蛋白去乙酰化酶mRNA水平变化

目的 探讨组蛋白去乙酰化酶(HDAC)抑制剂干预慢加急性肝衰竭(ACLF)小鼠肝组织组蛋白去乙酰化酶mRNA水平变化.方法 随机将小鼠分为7组,每组4只.A组为对照组,B组为模型组,C组为曲古抑菌素A(TSA)处理组,D组为大剂量正丁酸钠处理组,E组为小剂量正丁酸钠处理组,F组为大剂量吡咯烷二硫代氨基甲酸(PDTC)处...     

Th17细胞及其细胞因子与乙型肝炎后肝硬化肝组织炎症活动度相关性的研究

最新研究显示Th17细胞可能参与了非病毒特异性肝损伤的调控,然而至今尚无研究报道其是否参与了乙型肝炎后肝硬化的肝损伤。目的:探讨Th17细胞及其细胞因子与乙型肝炎后肝硬化肝组织炎症活动度的关系。方法:收集12例肝组织发生亚大块及以上坏死、12例肝组织炎症活动度为G2～G4的乙型肝炎后肝硬化患者以及11例健康人的肝组织和外周血标本,以免疫组化方法检测肝组织IL-17的表达和分布,实时荧光定量PCR检测肝组织Th17型细胞因子IL-17 mRNA和Th17细胞特异性转录因子RORγt mRNA的表达,ELISA方法检测血清IL-17水平。结果:亚大块坏死组肝组织IL-17阳性细胞浸润区域较G2～G4炎症组广泛,健康对照组仅有少量IL-17阳性细胞浸润。亚大块坏死组肝组织IL-17、RORγt mRNA表达和血清IL-17水平均明显高于G2～G4炎症组(P=0.0226,P=0.0531,P=0.0171).G2～G4炎症组又高于健康对照组(P=0.0289,P=0.0005,P=0.0160)。肝内IL-17 mRNA表达和血清IL-7水平与肝组织炎症活动度呈正相关(r_s=0.686,P0.0001;r_s=0.767,P0.0001)。结论:肝内和血清Th17型细胞因子IL-17水平与乙型肝炎后肝硬化肝组织炎症的严重程度呈正相关,Th17细胞及其细胞因子参与了乙型肝炎后肝硬化的肝损伤。     

乙型肝炎病毒相关慢加急性肝衰竭患者肝组织中环氧化酶-2和过氧化物酶体增殖物激活受体γ的表达

目的 研究HBV相关慢加急性肝衰竭(ACLF)患者肝组织的环氧化酶-2(COX-2)和过氧化物酶体增殖物激活受体γ(PPAR γ)的表达情况及其与临床指标的相关性.方法 选取ACLF患者35例,HBV相关慢性肝功能衰竭(CLF)患者35例,慢性乙型肝炎(CHB)患者27例及正常对照(NC)者15例,检测患者生物化学指标及HBV DNA水平;免疫组织化学法行肝组织COX-2和PPAR γ染色,检测并分析各组患者肝组织中COX-2和PPAR γ的表达水平及其与临床资料的相关性.多组间比较用Kruskal-Wallis检验,两组间比较用Mann Whitney非参数U检验,双变量的相关分析用Spearman相关分析.结果 ACLF组与CLF组、CHB组和NC组相比,AST、总胆红素、胆固醇、凝血酶原时间、国际标准化比率、纤维蛋白原、终末期肝病模型(MELD)评分差异有统计学意义(P值均<0.01).COX-2染色在肝细胞质呈阳性着色,PPAR γ染色以核阳性为主,亦可见细胞质阳性着色.ACLF、CLF、CHB和NC组肝组织COX-2表达水平评分分别为(4.77±0.95)分、(4.30±1.18)分、(4.65±0.70)分和(2.31±1.12)分,ACLF、CLF和CHB组表达水平高于NC组,差异有统计学意义(z值分别为-5.18、-4.50和-5.32,P值均<0.01);ACLF组高于CLF组,差异有统计学意义(z=-1.98,P<0.05).PPAR γ表达水平评分在ACLF、CLF、CHB和NC组分别为(6.23±1.78)分、(5.34±1.68)分、(5.90±1.06)分和(3.57±1.91)分,ACLF组高于其他各组,与CLF组及NC组相比,差异有统计学意义(z值分别为-2.62和-4.28,P值均<0.01).ACLF组的COX-2表达水平与MELD评分呈正相关(r=0.337,P<0.05),PPAR γ表达水平与HBV DNA水平的对数值呈正相关(r=0.348,P<0.05).结论 COX-2能反映肝脏炎症程度及肝脏损害程度,PPAR γ在慢性HBV感染时上调,并且炎症越明显,上调的幅度越大,可能是肝脏炎症时的保护机制.

Abstract：

Objective To study the expressions of cyclooxygenase-2 (COX-2) and Peroxisome proliferator-activated receptor gamma (PPAR γ) in liver of patients with hepatitis B virus (HBV) related acute-on-chronic liver failure (ACLF) and their correlation with clinical parameters. Methods 35 patients with ACLF, 35 patients with HBV related chronic liver failure (CLF), 27 patients with chronic hepatitis B (CHB) and 15 normal control were enrolled to study the expressions of COX-2 and PPAR γ in the liver tissues by immunohistochemical staining, and to analyze the correlation of the COX-2 and PPAR γ levels in liver tissues with clinical parameters. Results COX-2 was distinctly expressed in the cytoplasm of the hepatocytes, but PPAR γ was mostly expressed in the nuclei of the hepatocytes and also could be seen in the cytoplasm. The expressions of COX-2 in the liver of ACLF, CLF and CHB groups increased significantly as compared with NC group (z = -5.18, -4.50, -5.32, P < 0.01). The levels of COX-2 in ACLF livers also increased evidently as compared with CLF groups (z = -1.98, P < 0.05). The expression levels of PPAR γ in ACLF liver tissues were much higher than the other three groups, and statistical significances existed between ACLF group and the other two groups (CLF, NC groups) (z = -2.62, -4.28, P < 0.01). In ACLF group, the expression of COX-2 correlated with MELD score (r = 0.337, P < 0.05) and the expression of PPAR γ correlated with HBV DNA load (r = 0.348, P < 0.05). Clinical data showed that the levels of AST, TBil, CHOL, PT, INR, FIB and MELD score in ACLF group were significantly different from that in CLF, CHB and NC groups. Conclusions COX-2 expressed in liver may be a marker to reflect the degree of inflammation and injury of liver tissue. The PPAR γ expression of liver could be increased during chronic HBV infection and may be a protective mechanism against liver injury..     

Augmenter of Liver Regeneration may be a Candidate for Prognosis of HBV Related Acute-on-Chronic Liver Failure as a Regenerative Marker

Background/Aims: To search for a new regenerative marker to estimate the prognosis of acute-on-chronic liver failure (ACLF). Methodology: The CCl4 induced liver regeneration models were prepared and observed the change of ALR, hepatocyte growth factor (HGF), proliferation cell nuclear antigen (PCNA) and pathology. Meanwhile the sera of patients with HBV related liver disease were collected to examine the changes of ALR level and the prognosis of patients with ACLF was followed up. Results: After CCl4 injection, serum ALR level rose firstly and then declined in the ensuing 12 hours to near-basal level (F=30.495, p<0.01). ALR level in the liver tissue showed an inverse pattern. The changes of PCNA, HGF and pathology showed a consistent trend with serum ALR level. Serum ALR level was higher in ACLF (n=20) and hepatocellular carcinoma (n=20) than in normal control (n=10) (2.68±1.95 vs. 0.74±0.31, p<0.01; 1.77±1.32 vs. 0.74±0.31, p=0.035). Serum ALR level of patients with ACLF was more significant in survival group (n=10) than in dead group (n=10) in early stage of disease (7.83±1.77 vs. 2.14±1.58, t=7.576, p<0.01). Conclusions: ALR level in serum may indicate hepatocyte proliferation or liver regeneration. High ALR level in serum in early stage of ACLF may mean a good prognosis.     

异硫氰酸苄酯对脑胶质瘤U-87MG细胞周期的影响及其机制

目的观察异硫氰酸苄酯（BITC）对人脑胶质瘤细胞系U一87MG细胞周期的阻滞作用,并探讨其机制。方法取对数期u．87MG细胞分别以2、5Ixmol／L的BITC处理,应用流式细胞仪检测细胞周期分布,Westernblot—ting法检测细胞周期相关蛋白CyclinB1、Cdc2、p21、Cdc25C及磷酸化Akt（p-Akt）蛋白表达,RT—PCR法检测CyclinB1、Cdc2、p21和Cdc25C的mRNA表达,报告基因技术检测NF—KB、缺氧诱导因子（HIF）和真核细胞翻译起始因子4E（eIF4E）的转录活性。结果BITC作用24h后,u一87MG细胞周期G2／M期比例显著升高,CyclinB1、Cdc2、Cdc25C、p-Akt蛋白表达下调,p21蛋白表达上升,CyclinB1、Cdc2、Cdc25C、p21mRNA表达均显著下降,NF—KB、HIF、elF4E转录活性均显著下降（P均〈0．05）。结论BITC对U．87MG细胞周期有阻滞作用,机制可能与抑制Akt／NF-KB信号转导路径,进而调节细胞周期相关基因表达有关;此为胶质瘤的细胞周期靶点治疗提供了新思路。     

Elevated serum prostaglandin E2 predicts the risk of infection in hepatitis B virus-related acute-on-chronic liver failure patients

Objective: To evaluate the serum Prostaglandin E2(PGE2) level in Acute-on-chronic liver failure(ACLF) and determine its predicative value for infection.Methods: From April 2014 to April 2015, ninety-one patients with hepatitis B virus and ACLF but without infection were enrolled into this prospective study that was carried out at our Hospital. Twenty patients with stable chronic hepatitis B were enrolled from the outpatient department and twenty healthy control subjects without any disease were enrolled from hospital staff. Serum PGE2 levels were determined using ELISA at enrollment. Clinical and laboratory parameters were collected. Receiver operating characteristic(ROC) curves were used to determine optimal cut-off values to predict infection.Results: Significantly higher PGE2 levels were found in patients with ACLF in comparison with healthy controls and patients with stable CHB(P 0.000 1). In ACLF patients, PGE2 levels were significantly higher in patients that eventually developed infection than those without this complication(P 0.000 1). ROC analysis showed that serum PGE2(area under the ROC curve, 0.83) could predict infection in patients with ACLF with sensitivity of 78.4% and specificity of 81.5% using a threshold of 141 pg/m L.Conclusions: Serum PGE2 is associated with the susceptibility to secondary infections for patients with ACLF. Increased PGE2 serum levels may serve as a potential biomarker for developing infections in ACLF patients.     

Role of HSP90, CDC37, and CRM1 as modulators of P16(INK4A) activity in rat liver carcinogenesis and human liver cancer

Current evidence indicates that neoplastic nodules induced in liver of Brown Norway (BN) rats genetically resistant to hepatocarcinogenesis are not prone to evolve into hepatocellular carcinoma. We show that BN rats subjected to diethylnitrosamine/2-acetylaminofluorene/partial hepatectomy treatment with a "resistant hepatocyte" protocol displayed higher number of glutathione-S-transferase 7-7(+) hepatocytes when compared with susceptible Fisher 344 (F344) rats, both during and at the end of 2-acetylaminofluorene treatment. However, DNA synthesis declined in BN but not F344 rats after completion of reparative growth. Upregulation of p16(INK4A), Hsp90, and Cdc37 genes; an increase in Cdc37-Cdk4 complexes; and a decrease in p16(INK4A)-Cdk4 complexes occurred in preneoplastic liver, nodules, and hepatocellular carcinoma of F344 rats. These parameters did not change significantly in BN rats. E2f4 was equally expressed in the lesions of both strains, but Crm1 expression and levels of E2f4-Crm1 complex were higher in F344 rats. Marked upregulation of P16(INK4A) was associated with moderate overexpression of HSP90, CDC37, E2F4, and CRM1 in human hepatocellular carcinomas with a better prognosis. In contrast, strong induction of HSP90, CDC37, and E2F4 was paralleled by P16(INK4A) downregulation and high levels of HSP90-CDK4 and CDC37-CDK4 complexes in hepatocellular carcinomas with poorer prognosis. CDC37 downregulation by small interfering RNA inhibited in vitro growth of HepG2 cells. In conclusion, our findings underline the role of Hsp90/Cdc37 and E2f4/Crm1 systems in the acquisition of a susceptible or resistant carcinogenic phenotype. The results also suggest that protection by CDC37 and CRM1 against growth restraint by P16(INK4A) influences the prognosis of human hepatocellular carcinoma.