门静脉高压症治疗的现状与展望

本文涉及的内容是作为肝硬变合并症的门静脉高压症,这是门静脉高压症最常见的原因,也是最难治疗的,其后果也最严重。综观现代医学发展史,恐怕没有哪种疾患有这么多种内科和外科治疗手段问世,据不完全统计也有多达百种。治疗方法的多样恰恰说明治疗效果不满意。肝移植的出现     

大鼠库普弗细胞分离培养方法的改良

目的建立一种稳定、高效的大鼠库普弗细胞(KCs)分离培养方法。方法采用大鼠肝脏离体链酶蛋白酶E消化和胶原酶循环灌注消化。低速离心去除肝细胞,Histodenz不连续密度梯度离心和选择性贴壁的方法分离KCs。采用ED2 CD163、兔抗大鼠溶酶体膜相关蛋白2(LAMP2)免疫细胞化学．latex-beads吞噬实验和电镜观察来鉴定 KCs。结果KCs得率为5×107个,活率为98％,经鉴定ED2阳性细胞大于98％,LAMP2阳性细胞大于99％,吞噬试验及电镜观察证明分离所得为KCs。结论改良的分离培养方法稳定、高效,为进一步的研究打下了基础。     

肝癌组织内类阿片肽LVV-血衍吗啡素-6的作用

目的:分析肝癌组织与正常肝组织中血衍吗啡素类肽含量的差异并探究其意义.方法:肝癌患者12例,利用弱酸洗涤法从肝癌细胞和肝细胞表面提取肽段,然后用高压液相(HPLC)和质谱分析这些肽段并进行差异比较,筛选肿瘤特异性肽段.用这些肽段(0,10-7,10-8,10-9,10-10,10-11,10-12mol/L)处理人肝癌细胞株HLE,采用MTT法、流式细胞仪法观察其对肝癌细胞的生长、细胞凋亡的影响.结果:从7例患者的肝癌细胞表面成功检测出一条天然的类阿片肽——Leu-Val-Val(LVV)-血衍吗啡素-6(Mr1160.76).不同浓度LVV-血衍吗啡素-6均对HLE细胞有抑制作用,并且当浓度为10-7和10-8mol/L时有显著意义(24h:10-7vs10-8:P=0.044,10-8vs10-9:P=0.047;48h:10-7vs10-8:P=0.031,10-8vs10-9:P=0.040).同时,LVV-血衍吗啡素-6可诱导HLE细胞发生凋亡(浓度为0,10-7,10-8,10-9mol/L时的凋亡率分别为0.38%±0.09%,20.23%±1.25%,12.64%±2.15%,1.65%±0.34%),而阿片受体阻断剂naloxone能够逆转这种抑制效应(以上对应浓度时凋亡率分别为0.41%±0.11%,1.23%±0.45%,0.98%±0.55%,1.34%±0.43%).两组在LVV-血衍吗啡素-6浓度为10-7和10-8mol/L时有显著差异(P<0.05).结论:LVV-血衍吗啡素-6为肝癌细胞的病理产物,可激活阿片受体对肝癌细胞产生抑制作用.     

他克莫司胶囊和缓释胶囊预防肝移植急性排斥反应的效果和安全性

Objective To evaluate the efficacy and safety of tacrolimus exposure in stable liver transplant recipients converted from FK506 twice a day to Advagraf (tacrolimus extended-release capsules) once daily. Methods This was an open-label, random, control and multi-center study.Eligible patients were 19 to 70 years of age, 6 months post-transplant with stable renal and hepatic function and receiving stable doses of tacrolimus twice a day for 2 weeks prior to enrollment. There were 86 patients in the experimental group and the control group, separately. The average age of experimental group and control group was 46 ± 10 and 49 ± 9, respectively. Patients in experimental group received Advagraf, once daily, and the dose was adjusted according to the drug concentration,and the drug concentration was between 2 to 10 μg/L. The control group given tacrolimus, twice daily, and the drug concentration was between 2 to 10 μg/L. Results The incidence of acute rejection reaction was 1.20 % and 1.18 % respectively in experimental group and control group, and the 95 %confidence interval was -3.25% ～3.31 % and -3.26% ～ 3.34 %, individually. There was 1 case of acute rejection reaction in experimental group and control group, respectively. The patient and organ survival rate was 100%. Sixteen adverse events occurred in 15 patients (17.65 %) of the experimental group, and 10 adverse events occurred in 10 patients (11.63 %) of control group. Severe adverse events relating to the test drug in experimental group occurred in 4 patients (4. 71 %). and 2 patients (2. 33) in control group.Conclision Clinical trials indicated that Advagraf has efficacy and safety profiles similar to those of tacrolimus. The drug is safe and may improve patient compliance.     

活体肝移植供体手术及术后管理(附22例临床分析)

目的探讨提高活体肝移植供体手术安全性和加强术后管理的措施。方法回顾性分析北京大学人民医院2003年2月至2009年6月22例活体肝移植供体的临床资料,总结供体术前评估、手术技术、术后并发症及随访情况。结果术前对供体进行三步法评估,术中采用超吸刀(CUSA)进行肝脏实质分离,术后定期随访。供肝移植物包括肝左外叶2例、左半肝6例、扩大左半肝1例、右半肝5例和扩大右半肝8例。1例在术后早期出现腹腔内出血,1例合并胸水,1例枕后皮下血肿,其他供体未出现严重并发症,术后肝功能迅速恢复正常,随访至今无供体死亡。结论活体肝移植供体手术复杂,严格术前评估,术中精细操作,术后加强随访是提高活体肝移植供体安全的重要保证。     

十二指肠腺癌89例临床分析

目的 探讨原发性十二指肠腺癌的诊断和根治术术式选择.方法 对1985-2009年经手术切除、病理证实的89例十二指肠腺癌患者的临床资料进行回顾性分析.结果 十二指肠腺癌发病率低,临床表现缺乏特异性,术前内镜确诊率为93%,胃肠X线气钡造影为90%,磁共振胰胆管造影为82%,B超为42%,MRVCT为70%.肿瘤位于十二指肠第2段占65%.48例行胰十二指肠切除,19例行十二指肠节段切除,切除术后患者5年生存率分别为47%和50%.结论 内镜和X线气钡双重造影是诊断十二指肠腺癌的主要检查方法.早期诊断和根治手术是提高切除率和疗效的主要途径.

Abstract：

Objective To evaluate the preoperative diagnosis and treatment of primary duodenal adenocarcinoma ( PDA). Methods Clinical data of 89 PDA cases undergoing laparotomy from 1985 to 2009 in three hospital in Beijing and Zhengzhou were analyzed retrospectively. Results The prevalence rate of PDA is comparatively low and its clinical manifestations are not specific. The correct diagnostic rate of auxiliary examination were 93% by endoscopy,90% by gastrointestinal X-ray air barium double radiography,82% by MRCP,42% by BUS,70% by CT/MRI. The most common location of the adenocarcinoma(65% ) was at the second portion of the duodenum. Among 67 PDA cases 48 cases underwent pancreatoduodenectomy, 19 cases received segmental deodenectomy. The 5 year's survival rate was 47% and 50% respectively. Conclusions Endoscope and X-ray radiography are mainstays for the diagnosis of PDA. Early diagnosis and radical operation are the key to improve the resection rate and the long term survival.     

重组腺病毒Ad-PD-L1转染供体小鼠树突状细胞对受体小鼠淋巴细胞激活的影响

目的:观察重组腺病毒载体Ad-PD-L1转染供体C57BL/6(H-2b)小鼠树突状细胞(DC)对受体DBA/2(H-2d)小鼠淋巴细胞增殖和激活的影响.方法:构建腺病毒穿梭质粒pShuttle-GFPCMV(-)-PD-L1和腺病毒骨架质粒pAdxsi-GFP-CMV-PD-L1,重组腺病毒Ad-PD-L1进行包装、扩增和纯化.分离培养供体C57BL/6小鼠的DC,分为3组:腺病毒载体Ad-PD-L1转染组(A组),空载体转染组(B组)和对照组(C组).Western blot检测转染后各组细胞PD-L1的表达.分离受体DBA/2小鼠的淋巴细胞,用羧基荧光素二醋酸琥珀酰亚胺酯(CFSE)标记,将供体C57BL/6小鼠的DC和受体DBA/2小鼠淋巴细胞混合培养,流式细胞仪观察淋巴细胞的增殖情况.结果:酶切及测序证实含PD-L1的重组腺病毒载体Ad-PD-L1构建成功.转染供体C57BL/6小鼠的DC后其PD-L1的表达升高37% (P<0.05),将转染了PD-L1的DC与受体DBA/2小鼠的淋巴细胞进行共培养.与对照组相比,PD-L1表达升高后,明显地抑制了淋巴细胞的增殖和活化.受体DBA/2小鼠的淋巴细胞增殖降低41% (P<0.01).结论:转染供体C57BL/6小鼠DC后通过共刺激通路PD-1/PD-L1抑制了受体DBA/2小鼠淋巴细胞的增殖和激活.     

他克莫司胶囊和缓释胶囊预防肝移植急性排斥反应的效果和安全性

Objective To evaluate the efficacy and safety of tacrolimus exposure in stable liver transplant recipients converted from FK506 twice a day to Advagraf (tacrolimus extended-release capsules) once daily. Methods This was an open-label, random, control and multi-center study.Eligible patients were 19 to 70 years of age, 6 months post-transplant with stable renal and hepatic function and receiving stable doses of tacrolimus twice a day for 2 weeks prior to enrollment. There were 86 patients in the experimental group and the control group, separately. The average age of experimental group and control group was 46 ± 10 and 49 ± 9, respectively. Patients in experimental group received Advagraf, once daily, and the dose was adjusted according to the drug concentration,and the drug concentration was between 2 to 10 μg/L. The control group given tacrolimus, twice daily, and the drug concentration was between 2 to 10 μg/L. Results The incidence of acute rejection reaction was 1.20 % and 1.18 % respectively in experimental group and control group, and the 95 %confidence interval was -3.25% ～3.31 % and -3.26% ～ 3.34 %, individually. There was 1 case of acute rejection reaction in experimental group and control group, respectively. The patient and organ survival rate was 100%. Sixteen adverse events occurred in 15 patients (17.65 %) of the experimental group, and 10 adverse events occurred in 10 patients (11.63 %) of control group. Severe adverse events relating to the test drug in experimental group occurred in 4 patients (4. 71 %). and 2 patients (2. 33) in control group.Conclision Clinical trials indicated that Advagraf has efficacy and safety profiles similar to those of tacrolimus. The drug is safe and may improve patient compliance.     

通过PD-1/PD-L1共刺激通路诱导小鼠胰岛移植免疫耐受的研究

目的 观察重组腺病毒Ad-PD-L1对诱导小鼠胰岛移植免疫耐受的作用.方法 酶切含有小鼠全长PD-L1 cDNA的pSport 1-mCD274质粒,亚克隆到穿梭质粒pShuale-GFP-CMV(-)上,再通过酶切、连接等方法构建腺病毒骨架质粒pAdxsi-GFP-CMV-PD-L1,转化DH5α感受态细菌,筛选阳性克隆.经酶切、测序鉴定正确,线性化后脂质体法转染293细胞进行包装、扩增,氯化铯密度梯度离心纯化病毒.链脲霉素(250 mg/kg)腹腔注射诱导C57BL/6(H-2b)小鼠糖尿病模型,将C57BL/6小鼠随机分为3组,A为对照组,B为Ad-EGFP处理组,C为Ad-PD-L1处理组,在进行胰岛移植的前1天,经尾静脉输注5×108 pfu的Ad-PD-L1.将DBA/2(H-2d)小鼠的胰岛移植在糖尿病小鼠肾被膜下.术后监测不同时间各组小鼠的血糖变化及胰岛移植物的存活时间,并观察混合淋巴细胞反应的特异性.结果 酶切及测序证实Ad-PD-L1构建成功.Ad-PD-L1在小鼠体内可以高效地表达PD-L1,Ad-PD-L1治疗组胰岛移植物的存活时间明显延长到27.63±3.51 d(P<0.01),混合淋巴细胞反应表明小鼠对供体反应特异性降低. 结论通过PD-L1/PD-1共刺激通路可以抑制T细胞的活化,从而延长了胰岛移植物的存活时间.     

复杂性腹腔感染的莫西沙星单药治疗

莫西沙星(拜复乐 )单药可有效治疗复杂性腹腔感染,这是不久前完成的亚太地区多家医院参与的代号为"Dragon"的一项Ⅲ期临床试验结果证实的.