西罗莫司对人肝癌裸鼠肝脏移植瘤生长的影响

Objective To study the effects of sirolimus (SRL) on the growth of transplanted human hepatocellular carcinoma (HCC) in nude mice. Methods HepG2 cells were Implanted into the liver of nude mice. The implanted mice were then treated with SRL and tacrolimus (FK506). The expression of vascular endothelial growth factor (VEGF) and proliferating cell nuclear antigen (PCNA) was detected by immunohistology, microvessel density (MVD) was counted by immunostaining with anti-CD34 antibody for endothelial cells. Tumor apoptosis was detected by terminal deoxynuclcotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay. Results The tumor weight was (352±38) mg, (683±53) mg and (675±45) mg in SRL, FKS06 and control group respectively. The tumor weight was significantly decreased in SRL group (P < 0.0 I), and there was no difference between FKS06 group and control group. The expression of VEGF and PCNA protein was remarkably down-regulated in SRL group compared to control group (P < 0.05), and it was not significantly different between FK506 group and control group (P > 0.05). Compared to the control group, MVD was significanly decreased in SRL group, and the apoptosis index of tumor cell was significantly higher in SRL group (P < 0.01). Conclusion SRL inhibits transplanted HCC tumor growth by reducing tumor angiogenesis, inhibiting tumor proliferation and inducing tumor apoptosis.     

西罗莫司对人肝癌裸鼠肝脏移植瘤生长的影响

Objective To study the effects of sirolimus (SRL) on the growth of transplanted human hepatocellular carcinoma (HCC) in nude mice. Methods HepG2 cells were Implanted into the liver of nude mice. The implanted mice were then treated with SRL and tacrolimus (FK506). The expression of vascular endothelial growth factor (VEGF) and proliferating cell nuclear antigen (PCNA) was detected by immunohistology, microvessel density (MVD) was counted by immunostaining with anti-CD34 antibody for endothelial cells. Tumor apoptosis was detected by terminal deoxynuclcotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay. Results The tumor weight was (352±38) mg, (683±53) mg and (675±45) mg in SRL, FKS06 and control group respectively. The tumor weight was significantly decreased in SRL group (P < 0.0 I), and there was no difference between FKS06 group and control group. The expression of VEGF and PCNA protein was remarkably down-regulated in SRL group compared to control group (P < 0.05), and it was not significantly different between FK506 group and control group (P > 0.05). Compared to the control group, MVD was significanly decreased in SRL group, and the apoptosis index of tumor cell was significantly higher in SRL group (P < 0.01). Conclusion SRL inhibits transplanted HCC tumor growth by reducing tumor angiogenesis, inhibiting tumor proliferation and inducing tumor apoptosis.     

肝移植病人服用替比夫定后并发肌病和周围神经病3例分析

目的 探讨肝移植病人服用替比夫定后并发肌病（myopathy,MP）和周围神经病（peripheral neuropathy,PNP）的诊治方法。 方法 回顾分析2008年6月至2009年6月中山大学附属第三医院3例肝移植病人服用替比夫定后并发MP和PNP的临床特点及诊治过程。 结果 应用替比夫定的肝移植病人MP和PNP发生率为11.5%（3/26）,出现症状时间平均为用药后11.7个月,诊断时间平均为出现症状后10.7个月,治疗后平均3个月临床症状基本消失。 结论 肝移植病人同时服用替比夫定和他克莫司时MP和PNP的发生率较高,应注意观察其相关症状,监测血肌酸磷酸激酶和肌红蛋白,以便早期诊断和治疗MP和PNP。     

改良大鼠腹腔内心脏移植模型80例复制体会

目的 总结复制改良大鼠腹腔异位心脏移植模型的体会.方法 健康远交系Wistar大鼠为供体,SD大鼠为受体,各80只,将供心移植于受体的腹腔内,供心的升主动脉及右肺动脉分别与受体的腹主动脉和下腔静脉做端侧吻合;术后第3、5、7天各取5例移植心脏行组织学检查.结果 成功建立75例大鼠异位心脏移植模型,供心缺血时间(28±5.5)min,手术成功率93.75%(75/80).大鼠移植后均呈急性排斥反应改变.结论 通过调整动脉吻合口与静脉吻合口管径比率进一步完善改良移植模型,可稳定诱导急性排斥反应发生.     

老年患者的肝移植

目的 探讨60岁以上老年患者施行肝移植术治疗的临床特点和疗效。 方法 回顾2003年10月—2005年10月收治的57例60岁以上老年肝移植患者的临床资料，将其分为肝癌组和良性肝病组进行对比分析。结果 肝癌组31例患者中，男28例，女3例，平均（63.5+3.2）岁，符合米兰（Milan）标准者有12例，占38.7%。良性肝病组26例患者中，男22例，女4例，平均年龄（64.8+3.6）岁。两组相比在年龄、性别比、术前HbsAg阳性率，术前其它系统病史、术前手术史、术中出血量、手术相关并发症、术后肺部感染发生率等方面比较差异均无显著性（P>0.05）。在术前肝功能child-pugh分级上肝癌组以A级和B级为主（90.4%），良性肝病组以C级为主（76.9%），差异有显著性（P<0.05）。在术后呼吸机支持时间上肝癌组要明显短于良性肝病组（P<0.05）。平均随访（27.9±11.6）个月，肝癌组中存活22例，死亡9例，其中5例死因为肝癌复发和转移，良性肝病组存活18例，死亡8例，主要死因为多器官功能衰竭（4例），两组术后1年的生存率分别为77.4%和69.2%，2年的的生存率分别为71%和69.2%，差异均无显著性（P>0.05）。符合和超过Milan标准的老年肝癌患者在术后1年和2年的生存率和无瘤生存率且差异无显著性(P>0.05)。结论 接受肝移植治疗的60岁以上老年肝癌患者和良性肝病患者的疗效相当，老年肝癌肝移植具有较低的术后复发和转移率，将Milan标准作为选择老年肝癌患者施行肝移植的标准可能过于严格。     

5-脂氧合酶在大鼠外科脑损伤模型脑组织中的表达及作用

目的 研究大鼠外科脑损伤(SBI)模型脑组织中5-脂氧合酶(5-LOX)表达的空间分布、细胞定位和随时间变化的规律,并探讨其在SBI发病中的可能作用机制.方法 将72只健康成年雄性SD大鼠随机分为假手术(Sham)组及SBI术后1 d(SBI-1d)组、3 d(SBI-3d)组、7 d(SBI-7d)组,每组18只.对SBI-1d、SBI-3d和SBI-7d组大鼠采用右侧额叶切除法建立SBI模型,Sham组大鼠只移除相应部位的颅骨,不伤及硬脑膜.采用干湿质量法测量损伤同侧、对侧脑组织的含水量;Garcia评分和杠杆平衡评分评估各组大鼠的神经行为学功能;免疫荧光化学染色确定5-LOX的空间分布及细胞定位;蛋白质印迹法检测5-LOX和核转录因子(NF)-κB的蛋白表达;生物化学法测定损伤区周围脑组织中超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量.结果 (1)与Sham组相比,SBI-1d和SBI-3d组大鼠损伤侧脑组织含水量增加(P＜0.05);SBI-1d、SBI-3d和SBI-Td组大鼠出现神经功能障碍(P＜0.01),SBI-Td组大鼠的Garcia评分较SBI-1d、SBI-3d组改善(P＜0.05).(2)5-LOX主要分布在损伤区周围的脑组织中,其中以神经元表达为主,星形胶质细胞和小胶质细胞也有表达.(3)与Sham组相比,SBI-1d和SBI-3d组大鼠损伤区周围脑组织中5-LOX、NF-κB表达增加(P＜0.05),而SBI-7d组两者表达低于SBI-1d和SBI-3d组(P＜0.05),其中5-LOX表达增加在SBI术后第1天最明显.(4)与Sham组相比,SBI-1d、SBI-3d和SBI-7d组大鼠损伤区周围脑组织中SOD活性降低(P＜0.05),SBI-1d和SBI-3d组MDA含量增加(P＜0.05).结论 SBI后,5-LOX主要表达于损伤区周围神经元,星形胶质细胞和小胶质细胞也有表达;且在SBI术后1d表达最多.5-LOX加重脑损伤可能与NF-κB表达增多以及氧化应激增加有关.     

地震后四肢开放伤口的治疗策略

【目的】探讨地震后四肢开放伤口的治疗策略。【方法】本组伤员包括16人23处地震后四肢开放伤口,采用清创封闭负压引流处理伤口,伤口清洁后采用直接缝合、游离植皮术、邻位皮瓣和带蒂皮瓣转移闭合伤口。截肢感染残端的处理采用尽早彻底敞开伤口,充分引流,伤口清洁后立刻缝合伤口,必要时缩短残端。同时根据伤口创面分泌物培养结果选用敏感抗生素,对伤员的心理创伤、贫血、脱水和并发症积极治疗。【结果】本组伤员包括16人23处四肢开放伤口除1例91岁伤员外全部愈合。【结论】地震后四肢开放伤口采用正确的治疗策略,可以加速伤口愈合,尽快恢复肢体功能。     

肝肾联合移植13例临床分析

目的 探讨肝肾联合移植的适应证、手术并发症及生存情况.方法 回顾性分析2003年10月至2008年12月施行的13例肝肾联合移植患者的临床资料,分析围手术期死亡率、并发症情况及生存情况.结果 13例肝肾联合移植患者围手术期死亡率30.8%(4/13).术中、术后腹腔出血4例(30.8%);肺部感染7例(53.8%);移植肾急性排斥反应1例(7.7%).本组随访4.4～60个月,中位数40个月.存活1年以上8例,2年以上6例,3年以上5例,4年以上3例,5年以上1例.肝肾联合移植前有1例患者经历肝移植(例2)和2例患者经历肾移植(例3、例4),例4患者于肝肾联合移植术后第29天死于肺部感染、多器官功能衰竭,例2和例3肝肾联合移植术后分别存活40 m、48 m.结论 肝肾联合移植是治疗终未期肝肾疾病的有效方法.肝/肾移植术后再行肝肾联合移植是可行的.

Abstract：

Objective To investigate the indications, complications and survival results of combined liver-kidney transplantation. Methods From Oct 2003 to Dec 2008, the clinical data of 13 patients who underwent combined liver-kidney transplantation (CLKTs) were retrosptiverly analyzed in our institution. The perioperative mortality rate, complications and the result of follow-up were analyzed.Results The perioperative mortality rate (within 30 days) was 30.8% (4/13). Postoperative complications included intrabdominal bleeding in 4 patients ( 30. 8% ); pulmonary infection in 7 patients (53.8%); acute renal rejection in one (7. 7% ). Survivors were followed up from 4.4 to 60 months, with the median time of 40 months. Eight patients have survived more than 1 year; six patients have survived more than 2 years; five of them have survived for more than 3 years; and three of them have survived for more than 4 years, with one surviving for more than 5 years. One patient had undergone liver transplantation ( case 2 ) and two patients had had kidney transplantations ( case 3 and case 4 ) before this CLKTs.Postoperatively case 4 died of pulmonary infection and multiple organ failure at day 29, while case 2 and case 4 survived respectively 40 m, 48 m after CLKTs. Conclusions CLKTs is an effective therapy for end-stage liver and kidney disease. CLKTs for patients with irreversible liver and renal insufficiency after initial liver transplantation or kidney transplantation was feasible.     

siRNA沉默fas基因减轻大鼠肝移植供肝冷保存和缺血再灌注损伤

目的：探讨针对fas的siRNA对大鼠原位肝移植供肝冷保存和缺血再灌注损伤的拮抗作用及机制。方法:（1）化学合成针对大鼠fas基因的3对siRNA，转染大鼠正常肝细胞(BRL细胞)，筛选抑制效果最高的siRNA序列。（2）对SD大鼠用水压注射法转染fas siRNA，48 h 后取肝，供肝冷保存2、4、6 h 后行细胞凋亡指数及fas表达检测。（3）对照组、fas siRNA组各25对SD雄性大鼠，供肝冷保存 3 h 后行原位肝移植,再灌注后1、3、6、12和 24 h 每组各处死5只大鼠，查血谷丙转氨酶(ALT)；取供肝查fas mRNA表达、Fas蛋白水平和细胞凋亡计数。结果:315位点的siRNA抑制BRL细胞fas基因效率最高。供肝冷保存实验中，fas　siRNA组冷保存2、4、6 h 的肝脏fas表达与细胞凋亡指数低于对照组（P<0.01）。大鼠肝移植复流后，fas siRNA组各检测点的fas基因表达、蛋白水平均明显低于、凋亡细胞少于、血清ALT低于对照组。结论:针对fas的siRNA对大鼠肝移植中的供肝冷保存和缺血再灌注损伤有一定的拮抗作用。     

大鼠部分肝切除术后Hedgehog信号分子的表达

目的: 探讨部分肝切除术后Hedgehog(Hh)信号分子的表达及其意义。方法: 切除肝脏左叶和中叶以建立SD大鼠部分肝切除术模型。18只雄性SD大鼠随机均分为3组:假手术组(A)、部分肝切除术组1(B)、部分肝切除术组2(C)。A、B组于术后24 h、C组于术后48 h取肝组织,用免疫组化染色法和Western blotting检测肝组织Ki-67、Sonic Hedgehog(Shh)、Indian Hedgehog(Ihh)和Glioblastoma-2(Gli-2)的表达。结果: HE染色显示B、C组可见肝细胞水肿,点状坏死。C组可见处于不同分裂期的细胞。B、C组Ki-67、Shh、Ihh和Gli-2的表达明显高于A组(P<0.01),C组Ki-67、Shh、Ihh及Gli-2的表达高于B组(P<0.01)。结论: 部分肝切除术后Hh信号通路被激活,可能参与促进肝脏修复再生。