乌司他丁对肝大部切除合并缺血再灌注损伤后肝再生和能量代谢的影响

目的探讨乌司他丁(ulinastatin,UTI)预处理对大鼠70%肝切除术合并缺血再灌注损伤后肝再生和能量代谢的影响。方法健康清洁级雄性SD大鼠120只随机分为单纯肝切除组(PH)、肝切除合并缺血再灌注组(PHIR)和乌司他丁组(UTI),每组40只。将各组大鼠肝左、中叶切除,残肝(右、尾叶)作不同处理:PH组不阻断保留肝血流;PHIR组阻断保留肝血流30min后恢复灌注;UTI组与PHIR组肝脏切除及保留肝断流处理相同,但于缺血前5min经尾静脉给予UTI 50 000U/kg。再灌注后1、6、12、24、48h分别取各组大鼠下腔静脉血、肝组织,测定血清ALT和AST,肝细胞凋亡指数(AI);24、48h残肝再生度、PCNA阳性率和ATP、ADP和AMP含量。结果 UTI组的血清ALT和AST活性、AI均低于PHIR组(P<0.05),肝再生度和PCNA阳性率均高于PHIR组(P<0.05),ATP含量和能量负荷较PHIR组明显升高(P<0.05)。结论乌司他丁预处理能促进对大鼠70%肝切除术合并缺血再灌注损伤后的肝再生,其机制与维持能量代谢有关。     

Polyclonal antibody preparation and expression in liver tissues of transactivated protein 5 of hepatitis C virus nonstructural 5A

Objective To prepare polyclonal antibody of transactivated protein 5 of hepatitis C virus nonstructural 5A (NA5ATP5) and to explore its expression in the liver tissues. Methods In Escherichia coil BL21, the prokaryotic expression vector pET32a(+)-NS5ATP5 was induced by isopropyl-β-D-thiogalactoside (IPTG), and it was analyzed with sodium dodecyl sulfate-polyaerylamide gel electrophoresis (SDS-PAGE) and Western blotting. And the purified protein was used to immunize the rabbit to prepare polyelonai antibody, with which we studied the function of NSSATP5 by determining the different liver tissues with the streptavidin-perosidase (SP) immunohistochemistry method. Results Recombinant NS5ATP5 (molecular weight: 65 kD) and polyclonal antibody were successfully prepared. NS5ATP5 expression in the liver of patients with chronic HCV infection was much higher than that of a normal person, and it was detected mainly in the cytoplasm. Conclusion The findings of the expression difference between HCV patients and normal people led to a novel diagnostic marker to detect HCV infection.     

原发性肝癌P-糖蛋白表达与肝动脉插管栓塞化疗疗效的关系及临床意义

目的检测肝癌细胞的多药耐药基因MDR1及编码产物P-gp(P-glycoprotein,P-gp)的表达并探讨其与原发性肝癌经肝动脉插管栓塞化疗(transcatheter arterial chemoembolization,TACE)疗效的关系及临床意义。方法选取2010年6月到2013年6月来我院接受手术治疗的108例原发性肝癌患者作为肝癌组研究对象,同时选取来我院行肝穿刺检查的50例健康者作为对照。采用实时荧光定量PCR测定各组肝组织中MDR1的mRNA水平,并采用免疫组织化学法检测手术切除后的肝癌组织P-gp的表达。根据P-gp表达水平将患者分为耐药组和非耐药组,分析P-gp表达水平与原发性肝癌术后TACE治疗效果的关系。结果与对照组比较,肝癌组患者肝脏组织中MDR1的mRNA表达水平显著升高(P<0.05)。其中肝癌组MDR1的mRNA/正常水平>2的患者76例,MDR1的mRNA/正常水平<2的患者32例。免疫组织化学结果显示P-gp呈棕黄色,主要表达于肝癌细胞的细胞膜表面。其中P-gp蛋白阴性患者(非耐药组)35例,阳性患者(耐药组)73例。非耐药组患者化疗有效率(74.28%)明显高于耐药组患者(43.28%)(P<0.05)。非耐药组1、2年累积生存率为77.14%和42.86%,明显高于耐药组(54.12%和27.40%,P<0.05)。结论原发性肝癌组织中MDR1编码产物P-gp异常过表达与肿瘤化疗耐药性相关,提示其可作为临床术后化疗用药的指导标志之一。     

门静脉高压症患者肝组织CD44、血清透明质酸和肝功能性血流量的变化

目的探讨乙肝肝硬化门静脉高压症(LCPH)患者肝窦内皮细胞功能与肝功能性血流量(FLPF)变化的关系。方法选择50例LCPH患者和10例同期手术的非肝硬化肝囊肿患者,应用免疫组化法检测肝组织CD44的表达水平,放免法检测血清透明质酸的含量,D-山梨醇清除率法检测FLPF。结果对照组肝组织CD44表达水平很低,阳性染色面积为(3.49±0.38)μm2,LCPH组则明显上调,达(41.38±11.88)μm2。对照组血清透明质酸含量为(48.15±14.39)μg/L,LCPH组升高,达(116.11±20.12)μg/L。对照组FLPF为(1 261.22±65.79)mL/min,LCPH组降低至(770.33±48.53)mL/min。结论 LCPH患者肝窦内皮细胞功能的变化与FLPF的降低密切相关。     

硒抗肝纤维化作用的实验研究

以综合法建立大鼠慢性肝损伤模型 ,观察硒对肝羟脯氨酸( Hydroxyproline,Hyp)含量及超微结构影响。结果 :补硒预防组肝 Hyp含量明显低于模型对照组 ( P <0 .0 5 ) ,超微结构显示肝组织内胶原原纤维亦明显减少 ,上述改变与秋水仙素组比较仍有差距。提示硒有一定抗纤维化作用。     

In situ absorption and metabolism of stilbene glycoside in rat

Stilbene glycoside (TSG) has been shown to have many beneficial properties. It is therefore essential to understand the absorption and metabolism of TSG in detail. We determined the recovery of TSG and its metabolites (TSG sulfate/glucuronides) in rat gastric contents, gastric mucosa, portal vein plasma, celiac arterial plasma, bile, and urine after administration of 15mg of TSG in 0.5mL physiological saline or incubation for 20min in situ in the stomach of rats. Within 20min, (64.0±9.8)% of the administered TSG disappeared from the stomach; later, TSG was recovered in both free and conjugated forms in plasma and bile, but not in urine. On the other hand, only free TSG was detected in the gastric contents and mucosa; it was also detected in the portal vein plasma as (48.1±3.5)% of the total TSG (all forms of TSG). However, the proportion of free TSG in the celiac arterial plasma and bile decreased to 4%-10%. In addition, the proportion of free TSG to total TSG in the liver microsome incubation mixture after TSG was incubated in liver microsome at 37℃ for 30min was very low [(10.6 ± 2.6)%]. These results indicate that TSG could be quickly absorbed from the rat stomach, conjugated in liver and excreted in bile. Such novel information would be helpful for the use of TSG as a beneficial natural product which may improve its proposed efficacy in preventing chronic diseases.     

经颈内静脉肝内门体分流术治疗门脉高压

评价经颈内静脉肝内门体分流术治疗门脉高压症的应用价值。方法采用ＲＵＰ－１００穿刺导管系统对１０例门脉高压患者经右颈内静脉宽刺，建立静脉脉和肝静脉之间肝内通道后置入血管同人支架。     

HGV与HBV、HCA重叠感染病例的研究

探讨庚肝病毒 ( HGV)与 HBV、HCV重叠感染状况。方法　采用套式逆转录聚合酶链反应 ( RT- n PCR)技术检测了 2 2例乙肝病毒感染者及 48例丙肝病毒感染者血清 HGVRNA。结果　 HGV与 HBV、HCV重叠感染率分别为 31 .8% ( 7/ 2 2 )、31 .3% ( 1 5/ 48) ,重叠感染病例与单纯 HBV、HCV感染者在 ALT、SBIL水平及疾病的临床分型上无统计学差异。结论　HGV与 HBV、HCV重叠感染的几率均等 ,HGV似不加重乙肝及丙肝患者的病情     

PKM2对肝内胆管癌细胞增殖及上皮间质化的影响

目的 检测M2-型丙酮酸激酶(pyruvate kinase M2,PKM2)在肝内胆管细胞癌(intrahepatic cholangiocarcinoma,ICC)组织中的表达情况,探讨PKM2对ICC细胞增殖及上皮间质化(EMT)的影响.方法 RT-PCR和免疫组化法检测ICC组织中PKM2的表达,慢病毒载体介导...