输尿管非结石性梗阻病因及诊断分析

目的提高对输尿管非结石性梗阻的诊断水平.方法回顾性分析近10年来收治的输尿管非结石性梗阻患者的病因及诊断方法.结果损伤性输尿管狭窄72例,输尿管癌22例,输尿管囊肿18例,输尿管结核13例,先天性肾盂输尿管交界处狭窄8例,非特异性输尿管炎7例,输尿管息肉6例.结论输尿管癌为中老年输尿管非结石性梗阻患者的首要病因,损伤性输尿管狭窄为青年患者的首要病因.通过病史和恰当的检测手段,可以明确病因.     

小鼠骨髓干细胞体外定向诱导为肝细胞样细胞的实验研究

目的来源于骨髓干细胞的有功能的干细胞,可能成为生物人工肝和肝细胞移植的细胞来源。我们建立恰当的体外诱导培养体系,促使骨髓干细胞转化为肝细胞。方法获取骨髓干细胞,建立以FGF、HGF、OSM、EGF为主的细胞诱导培养体系。在细胞分化过程中,观察细胞形态和数量的变化,RT-PCR检测基因表达的变化,免疫荧光染色技术在蛋白水平检测ALB和CK18表达情况。PAS染色检测其糖代谢功能。结果在诱导过程中,多极性的肝细胞样细胞在12 d内可以观察到,且其随着细胞分化过程逐渐增多、集落增大。诱导细胞在7 d内表达AFPmRNA并维持到第21天,但后期表达减弱;在7天内表达ALBmRNA和CK18mRNA,随着分化时间的延长,其表达不断增强;在14 d内表达TTRmRNA,随着分化时间的延长,其表达不断增强。免疫荧光染色进行定位:在诱导组,诱导21 d的细胞表达ALB和CK18,其中ALB表达于胞浆和胞膜,而CK18表达于胞浆。糖原染色发现诱导组中,细胞胞浆内出现大小不等的红染的糖原颗粒。结论我们建立的以细胞因子FGF、HGF、OSM、EGF为主的细胞诱导培养体系是有效的,通过诱导骨髓干细胞,我们获得了有部分肝细胞形态结构和功能的肝细胞样细胞。     

血管内皮生长因子与膀胱癌研究进展

血管内皮生长因子 (VEGF)是肿瘤血管形成的关键因子 ,在肿瘤的发生发展过程中起重要作用。本文就VEGF及其VEGF在膀胱癌中的研究和应用作一综述 ,以便寻求膀胱癌诊断、监测、估计预后及治疗的新手段。     

小鼠骨髓基质干细胞体外转化为肝细胞最佳诱导体系的筛选及验证

目的:采用均匀设计法筛选及验证小鼠骨髓基质干细胞体外转化为肝细胞的最佳诱导培养体系.方法:获取小鼠骨髓基质干细胞,根据均匀设计法分8组进行体外诱导实验.通过流式细胞术检测各组ALB及CK18的阳性表达率,通过逐步回归分析法确立最佳细胞因子组合及浓度.从基因水平、蛋白水平以及细胞合成代谢功能检测.证实诱导的细胞为有功能的肝细胞.结果:FGF取35 μg/L,OSM取30μg/L时,ALB及CK18的阳性细胞达到最高值.采用该最佳诱导体系诱导过程中可检测到细胞表达ALBmRNA、CK18 mRNA、AFP mRNA、TTRmRNA:以及ALB、CK18蛋白表达:第21天最佳体系诱导组ALB阳性细胞的比例为82.83%±9.03%.CK18阳性细胞的比例为74.79%±8.41%.诱导培养过程中细胞分泌尿素及白蛋白,且随诱导时间的延长而增强.结论:均匀设计法可有效进行最佳诱导体系的筛选,以35 μg/LFGF、30μg/LOSM为主的诱导培养体系,可以有效地促进骨髓基质干细胞体外定向转化为有功能的肝细胞.     

肝再生发生及其相关机制的研究进展

肝脏再生是指因手术、创伤、中毒、感染、坏死等致部分肝细胞功能丧失后,肝细胞重新修复的过程.对于肝再生的发生及其内在机制的认识和研究,大致可分为三个阶段:(1)最初认为可能存在一种特定的激素类物质决定肝再生的过程.     

训练免疫在相关疾病中的研究进展

训练免疫（trained immunity,TI）是不依赖T、B细胞的,区别于传统固有免疫特征的一种记忆性固有免疫应答,在机体初次感染后对相同或不同病原体再次感染表现高反应性。训练免疫作为一种新兴的观点,具体机制涉及固有免疫、代谢及表观遗传之间的交互作用。基于此,本文总结了脂多糖（lipopolysaccharide,LPS）、卡介苗（bacille calmette?guerin,BCG）、激素等刺激对单核/巨噬细胞、自然杀伤细胞等先天免疫细胞以及非经典免疫细胞的表观遗传和代谢修饰的影响,并探讨了调节训练免疫在炎症、神经系统疾病以及新冠肺炎等方面的作用。     

体外生物人工肝支持系统治疗肝功能衰竭的系统评价

背景：生物人工肝支持系统由生物反应器和细胞材料组成,治疗过程中可部分替代肝脏的主要功能如解毒、合成、分泌和生物转化等功能。目的：回顾和评价体外生物人工肝支持系统近15年来治疗肝功能衰竭的疗效。方法：检索1995至2009年PubMed和Cochrane数据库有关生物人工肝支持系统的临床研究。被纳入的研究类型包括随机对照试验、临床对照试验和病例报告。结果与结论：共纳入31项研究,在生物人工肝支持系统治疗之后,患者生化指标呈现好转趋势,且大部分患者神经系统症状均得到改善,但多数患者生存率未见明显改善。说明尽管生物人工肝支持系统被证明具有一定治疗效果,但今后仍然有许多方面需要改进。     

中晚期胆囊癌的外科治疗策略:附17例报告

目的 探讨中晚期胆囊癌(NevinⅢ～Ⅴ期)的外科治疗方法,进一步提高患者存活时间.方法 结合文献,联系17例中晚期胆囊癌的围手术期临床资料及随访数据,着重从患者术前诊断、手术治疗策略、术后并发症及存活时间等方面进行探讨.结果 通过术前多种影像学资料相互印证,可以诊断中晚期胆囊癌并对分期做出判断,但是不能避免误诊;手术治疗中晚期胆囊癌的要点在于手术范围的确定,特别是淋巴结彻底清扫的程度,本组13a淋巴结阳性率35.3%;8淋巴结阳性率23.5%,说明为了尽量保证肿瘤无残留必要时可适当扩大手术范围;手术后并发症主要包括腹腔感染、胆瘘和麻痹性肠梗阻.结论 中晚期胆囊癌外科治疗需要适当手术范围,术中肿瘤无残留可以使患者存活较长时间.

Abstract：

Objective To explore the optimal surgical treatment strategy of advanced gallbladder carcinoma (Nevin Ⅲ - Ⅴ ), with an aim to prolong patients' overall survival. Methods 17 patients with advanced gallbladder carcinoma were reviewed. Their preoperative diagnosis, surgical treatment, complications and survival time were studied. Results The diagnosis of advanced gallbladder carcinoma was done using different medical imaging techniques, but incorrect diagnosis still happened. There is a wide range of surgical treatment for advanced gallbladder carcinoma. Controversy still exists as whether lymph node resection should be done. In our patients, 35.3% of the 13a lymph nodes and 23. 5% of the 8 lymph nodes were positive for metastasis, which showed that lymph node resection should be carried out. Extended surgery was sometime required to ensure a R0 resection.The main complications of surgery were intraabdominal infection, bile leakage and paralytic ileus.Conclusion An aggressive surgical approach for advanced gallbladder carcinoma is required to ensure a R0 resection, which contributed to better overall survival.     

兔慢性肝损害急性肝衰竭动物模型的建立

目的 建立新西兰兔的慢加急性肝衰竭(ACLF)动物模型.方法 将75只新西兰兔随机分为实验组(n=70)与对照组(n=5),实验组采用四氯化碳(CCl4)腹腔注射建立兔代偿性肝纤维化模型,每周2次,通过体质量及谷丙转氨酶(ALT)/谷草转氨酶(AST)改变调整药物剂量,共10周,获得48只肝纤维化新西兰兔,在此基础上将动物随机分为4组(n=12),Ⅰ组:继续腹腔注射CCl4,Ⅱ组:静脉注射D-氨基半乳糖(D-Gal)0.65 g/kg,Ⅲ组:静脉注射D-Gal 0.70 g/kg,Ⅳ组:静脉注射D-氨基半乳糖0.75 g/kg,观察各组动物的一般情况、生化指标及病理改变.结果 与对照组比较,肝纤维化实验组兔10周时ALT、AST、ALB、γ-谷氨酰转肽酶(GGT)、透明质酸(HA)、层粘连蛋白(LN)、Ⅲ型前胶原(PC-Ⅲ)差异均有统计学意义(P＜0.05),可观察到肝纤维化的病理表现,出现典型的假小叶,在肝纤维化基础给予D-Gal后,Ⅰ组动物全部存活,生化指标轻度改变;Ⅱ组动物死亡率为33.3%(4/12),生化指标仅出现一过性的改变;Ⅲ组动物死亡率为83.3%(10/12),平均存活时间为(53.00±25.69)h,给药后12 h生化指标及临床表现出现改变,48 h达到高峰,病理显示肝脏大块坏死;Ⅳ组动物均死于肝衰竭,平均存活时间为(32.70±17.46)h,肝损害出现时间早,损伤剧烈.结论 对CCl4诱导的肝纤维化新西兰兔给予D-Gal急性攻击可建立ACLF模型.其中给予D-Gal 0.70 g/kg的模型稳定性好,能较大程度上的模拟临床上ACLF的病理生理过程.

Abstract：

Objective To establish an ideal animal model of acute-on-chronic liver failure (ACLF) in New Zealand white rabbits in order to provide a large animal model for further researches.Methods Totally 75 New Zealand rabbits were randomly divided into experimental group (n =70) and control group (n = 5 ). Rabbits in the experimental group were injected with CCl4 into the abdominal cavity twice every week and the doses of CCl4 were modified according to the index of liver function and the body weight, whereas those in the control group were treated with the same volume of saline. At the 10th week,48 New Zealand rabbits with hepatic fibrosis were randomly assigned to 4 groups and injected with CCl4 as before, D-Gal at a dose of 0. 65 g/kg body weight (BW), 0. 70 g/kg BW and 0. 75 g/kg BW, respectively. By observing and comparing the general state, survival time, biochemical indexes, and the histopathology, a method of establishing a stable animal model of acute hepatic failure was found. Results As compared with those in control group, the levels of ALT, AST, GGT, HA, LN and PC-Ⅲ in the experiment group were increased significantly, while the level of ALB was decreased at the end of 10 weeks. Typical features of hepatic fibrosis and the formation of pseudo-lobules were observed at the end of 10 weeks. After treatment with D-Gal, all rabbits in group Ⅰ survived with minimal changes in liver function tests. In group Ⅱ , there was a temporary hepatic injury, but no hepatic coma. Four of the 12 rabbits died (33. 3% ). In group Ⅲ , biochemical indexes changed obviously 12 h after the administration and hepatic injury reached its peak after 48 h. Ten of 12 rabbits were died of severe hepatic failure with a survival time of ( 53. 00 ± 25. 69) h. Histology of liver section revealed massive necrosis in nodules. In group Ⅳ , hepatic injury occurred early and severely. All the rabbits died of severe hepatic failure with a survival time of (32. 70 ± 17. 46) h. Conclusion The experimental model of ACLF could be established by injected with D-Gal in New Zealand rabbits with hepatic fibrosis, induced by CCl4 intraperitoneal injection for 10 weeks.The one induced by 0. 70 g/kg of D-galactosamine was more stable and showed similar clinical pathophysiological changes in human beings. So it can be a good experimental platform for studies of ACLF.     

永生化人肝细胞系的建立及其生物学特性

目的 建立一种由慢病毒载体表达人端粒酶反转录酶亚单位,具有原代肝细胞的特性的永生化肝细胞系.方法 采用离体两步胶原酶灌注法无菌条件下分离肝细胞,构建慢病毒载体并包装高滴度慢病毒,肝细胞被采用表达永生化基因的慢病毒转染.通过筛选功能基因阳性表达克隆建立永生化肝细胞系,倒置相差显微镜观察形态学变化,免疫组化检测细胞角蛋白18(CK18)表达,RT-PCR及Westernblot检测功能基因及蛋白表达.结果 采用功能基因阳性表达筛选建立了永生化肝细胞系,其可连续扩增传代,具有原代肝细胞的特性并未呈现致瘤性,揭示出肝脏实质细胞的形态学特征并且表达肝细胞主要的功能基因标志.结论 通过功能基因阳性表达筛选成功建立具有原代肝细胞特性的永生化肝细胞系.