趋化因子受体CXCR4在结肠直肠癌中的表达及意义

目的：研究趋化因子受体CXCR4在结肠直肠癌中的表达及其临床意义。方法：采用RT-PCR法检测8种结肠直肠癌细胞株中CXCR4mRNA表达,应用蛋白印迹法检测细胞株中CXCR4蛋白表达。应用免疫组化染色法、结合组织芯片检测正常结肠直肠黏膜组织54例、结肠直肠癌组织49例中之CXCR4蛋白表达情况,并探讨其与结肠直肠癌临床病理特征的关系。结果：在8种结肠直肠癌细胞株中,CXCR4基因在HT29中表达水平最高。COL0205及SW480次之,HCTll6最低。CXCR4蛋白在结肠直肠癌组织中的表达高于结肠直肠正常黏膜组织,差异具有统计学意义（P=0．000）;在伴发转移的结肠直肠癌组织中,其表达比未发生转移者增强,差异具有统计学意义（P=0．024）。结论：CXCR4表达与结肠直肠癌转移存在密切关系,可能成为结肠直肠癌转移潜在治疗靶点。     

低位直肠癌套入式结肠直肠黏膜吻合保肛术102例

目的 探讨套入式结肠直肠黏膜吻合术治疗低位直肠癌保留肛门括约肌功能的安全性和可行性。方法 对102例中下段直肠癌行经腹肛门切除套入式结肠直肠黏膜吻合术治疗的病例进行回顾性总结。结果 102例患者术后未发生吻合口瘘和吻合口狭窄。术后早期排便次数较多,6～12次／d,口服止泻药物,可达到控制排便次数,术后12～18周时肛门排便功能基本恢复到正常。患者随访率89．2％(91／102),平均随访4．8年。局部复发率为5．4％(5／91),肝转移率为13．1％(12／91)。术后3年生存率为86．9％(60／68)。术后5年生存率为70．7％(29／41)。结论 套入式结肠直肠黏膜吻合术既可避免结肠造口,又可有效防止吻合口瘘,是一种安全有效的直肠癌保肛术式。     

Vpr对人结肠癌细胞的抑制作用及其机制

目的 探讨人类免疫缺陷病毒1型( HIV-1)病毒蛋白r(Vpr)对人结肠癌细胞HCT-8的抑制作用及其机制.方法 将细胞分为空白对照组、空载体转染组和Vpr转染组.空白对照组:细胞不予干预;空载体转染组:对数生长期的细胞加入不同感染复数(MOI)的空载体腺病毒;Vpr转染组:加入不同MOI的含有HIV1-Vpr基因的重组腺病毒(Adv-Vpr).应用MTT比色法检测细胞增殖活性,流式细胞仪检测细胞周期、细胞凋亡及线粒体膜电位,Western blot法检测细胞凋亡相关蛋白的表达.多组间均数比较采用单因素方差分析,两两比较采用q检验;或采用双因素方差分析,组内比较采用t检验.结果 Vpr显著抑制人结肠癌细胞HCT-8增殖,Adv-Vpr转染72 h后(MOI=200),Vpr转染组细胞MTT值为1.03±0.04,与空载体转染组(2.46 +0.15)及空白对照组(2.51±0.14)比较,差异有统计学意义(F=144.6,P＜0.05);Adv-Vpr转染48 h后(MOI=200),Vpr转染组处于G2/M期的细胞比例及线粒体膜电位下降的细胞比例增加,分别为37.31％±5.90％和32.07％±5.64％,与空载体转染组(18.30％±6.04％、3.32％±0.79％)及空白对照组916.66％±3.51％、2.76％±1.43％)比较,差异有统计学意义(F=10.08,64.45,P＜0.05).Adv-Vpr转染72 h后(MOI=200),Vpr转染组细胞凋亡率为37.62％±6.48％,与空载体转染组(3.44％±1.11％)及空白对照组(2.93％±1.07％)比较,差异有统计学意义(F=122.4,P＜0.05).Adv-Vpr处理48 h后(MOI=200),Westem blot检测发现Vpr导致了Caspase-9、Caspase-3剪切为活性片段,p-Chk1-S345磷酸化增加,而Fas、Fas-L、ERK1及ERK2表达未见上调.结论 Vpr在体外能够有效抑制结肠癌细胞株HCT-8增殖,并诱导其细胞周期G2期阻滞及凋亡,其机制分别与DNA损伤信号通路激活及线粒体凋亡通路启动有关.Vpr在结肠癌的治疗中具有潜在的应用前景.     

腹腔镜胃上部癌根治术残胃空肠双通道重建

目的 探讨腹腔镜胃上部癌根治术残胃空肠双通道重建的价值.方法 对25例胃上部癌患者行腹腔镜近端胃大部根治性切除,保留远端残胃,行空肠残胃侧侧吻合、空肠空肠端侧吻合的双通道消化道罩=建术.结果 手术平均时间(240±35)min,切缘阴性,平均清扫淋巴结(22±5)枚,无吻合口漏及狭窄、梗阻等并发症.术后均行钡餐造影,显示大部分钡剂直接进入空肠,少部分经过残胃.十二指肠进入空肠,并在残胃中滞留30～60min,无钡剂反流入食管.随访4～18个月,患者无明显反流性食管炎表现,近期生活质量满意.结论 残胃空肠双通道重建术清扫、切除范围合理,残胃有一定储袋作用,能较好地预防反流性食管炎和倾倒综合征;保留了十二指肠径路,提高了患者生活质量,是胃上部癌根治术较理想的消化道重建方式.     

放大肠镜在重症急性胰腺炎早期的结肠黏膜损害中的诊断价值

目的:探索放大肠镜在重症急性胰腺炎(SAP)早期肠黏膜屏障功能障碍导致的结肠黏膜损伤诊断中的价值。方法:对35例早期SAP病人进行放大肠镜检查,观察结肠黏膜有无损害,并取活检进行病理学光镜和电镜检查,同时对其进行APACHEⅡ评分、胃肠功能评分、胰周组织感染发生率统计。结果:71.4%(25/35)的早期SAP病人在放大肠镜检查中发现有结肠黏膜损伤,包括黏膜水肿、出血、糜烂。77.1%(27/35)的病人活检组织病理检查发现异常,包括黏膜上皮脱落、紧密连接缩短、细胞间分离。放大肠镜发现结肠黏膜损伤的病人其病理检查异常率、APACHEⅡ评分、胃肠功能评分、胰周感染发生率均高于未发现结肠黏膜损伤的病人。结论:对早期SAP病人进行放大肠镜检查,能够评估病人疾病的严重程度,预测病程进展中发生胰周感染的概率。     

手辅式腹腔镜右半结肠癌根治术在老年肥胖患者中的应用

目的评价手辅式腹腔镜右半结肠癌根治术在老年肥胖患者中的应用价值。方法回顾分析我院行手辅式腹腔镜右半结肠癌根治术的老年肥胖患者20例，以及同期开腹右半结肠癌根治术患者25例，并进行两组比较。结果两组均无手术死亡病例，均行一期吻合；手辅式腹腔镜组术中出血量少、术后体温恢复快、排气早、术后住院时间短、与开腹组相比差异有显著性意义（P〈0．01）；手辅式腹腔镜组手术时间与开腹组差异无显著性意义；手辅式腹腔镜组手术切除标本长度、清扫淋巴结总数及术后并发症与开腹组比较均无显著性差异。结论手辅式腹腔镜右半结肠癌根治术应用于老年肥胖患者，具侵袭性小、操作安全可行、术后恢复快等优点，是一种较为理想的微创手术方式。     

反义细胞外信号调节激酶-2基因治疗移植物动脉血管病内膜病变

目的 观察移植物动脉血管病(TA)的内膜病变机制和反义细胞外信号调节激酶2基因腺病毒载体(Adanti-ERK2)基因治疗的效果.方法 建立Brown-Norway(BN)-Lewis移植物动脉血管病模型,分为同系组、Control组、LacZ组和Adanti-ERK2组(给予5×109 pfu Adanti-ERK2基因治疗),每组各6例.术后60 d检测各组内膜病变和血管腔内膜/(内膜+中膜)比,α-肌动蛋白(α-actin)和血小板源性生长因子-BB(PDGF-BB)染色检测移植动脉平滑肌细胞(VSMCs)增殖和分泌功能,评估移植动脉新生毛细血管情况并检测移植动脉中环氧化酶-2(COX-2)的表达.结果　术后60 d同系组内膜无异常,Control组和LacZ组典型内膜增殖改变,Adanti-ERK2组内膜病变较轻;内膜/(内膜+中膜)比各组分别为7.6%、81.4%、85.9%、15.9%;α-actin阳性细胞(内膜平滑肌细胞)每视野计数各组分别为0、71.3±9.2、76.4±11.3、34.8±5.3;PDGF-BB阳性细胞每视野计数各组分别为0.9±0.5、28.4±3.4、29.1±3.2、8.6±1.7;移植动脉中膜和内膜新生毛细血管检测各组分别无、丰富、丰富、少量;COX-2新生血管阳性细胞计数各组分别为0、36.3±8.3、40.9±9.2、10.4±3.9.Adanti-ERK2组与其他组别间比较,差异有统计学意义(P＜0.05).结论　内膜增生,血管腔缩窄,PDGF-BB诱导内膜平滑肌细胞募集分化并激发血管新生是TA重要病理生理环节,AdantiERK2基因治疗可有效干预各发病环节,达到治疗效果.

Abstract：

Objective To explore the mechanisms of intimal injury underlying transplant arteriosclerosis (TA) and to clarify the treatment effect of adenovirus-mediated anti-extracellular signal regulated kinase 2 (Adanti-ERK2) gene therapy on TA. Methods The Brown-Norway (BN)-Lewis TA model was employed. According to different gene therapy, the recipients were divided into isograft group, control group, LacZ group, which were used as control, and Adanti-ERK2 group (5 × 109 pfu Adanti-ERK2 was transferred into the graft before transplant, 6 cases in each group). The grafts were harvested on the day60 post-transplantation to evaluate the intimal injury and calculate the ratio of intima/( intima + media). The staining of α-actin and PDGF-BB was performed to analyze proliferation and secretion of vascular smooth muscle cells (VSMCs). The angingenesis was evaluated and the cyclooxygenase-2 (COX-2) staining was detected. Results The intima was normal in the isograft group but a typical intimal proliferation was observed in the control group and the LacZ group. A mild intima injury was obtained in the Adanti-ERK2 group. The ratio of intima/( intima + media) was 7.6%, 81.4%, 85.9% and 15.9% in isograft, control, LacZ and Adanti-ERK2 groups, respectively. The α-actin staining-positive cells, which indicated VSMCs, were counted per vision-field as 0, 71.3 ± 9.2, 76. 4 ± 11.3 and 34. 8 ± 5.3, respectively. The platelet derived growth factor-BB (PDGF-BB) positive cells were counted as 0. 9 ± 0. 5, 28.4 ± 3.4,29. 1 ± 3.2 and 8.6 ± 1.7, respectively. The angiogenesis was detected as none, abundant, abundant and few, and COX-2 positive cells were counted as 0, 36. 3 ± 8. 3, 40. 9 ± 9. 2 and 10. 4 ± 3.9, respectively (P ＜ 0. 05 between Adanti-ERK2 group and other groups). Conclusion Intimal proliferation, luminal narrow, VSMCs recruitment and differentiation induced by PDGF-BB in intima and the following angiogenesis are the important pathophysiological process of TA. Adanti-ERK2 gene therapy modulates the process and then ameliorates TA.     

Metabolic Dysfunction-Associated Fatty Liver Disease Predicts Long-term Mortality and Cardiovascular Disease

Background/AimsWe investigated the effect of metabolic dysfunction-associated fatty liver disease (MAFLD) on future mortality and cardiovascular disease (CVD) using a prospective community-based cohort study.MethodsIndividuals from two community-based cohorts who were 40 to 70 years old were prospectively followed for 16 years. MAFLD was defined as a high fatty liver index (FLI ≥60) plus one of the following conditions overweight/obesity (body mass index ≥23 kg/m²), type 2 diabetes mellitus, or ≥2 metabolic risk abnormalities. Nonalcoholic fatty liver disease (NAFLD) was defined as FLI ≥60 without any secondary cause of hepatic steatosis.ResultsAmong 8,919 subjects (age 52.2±8.9 years, 47.7% of males), 1,509 (16.9%) had MAFLD. During the median follow-up of 15.7 years, MAFLD independently predicted overall mortality after adjustment for confounders (hazard ratio [HR], 1.33; 95% confidence interval [CI], 1.05 to 1.69) but NAFLD did not (HR, 1.20; 95% CI, 0.94 to 1.53). MAFLD also predicted CVD after adjustment for age, sex, and body mass index (HR, 1.35; 95% CI, 1.13 to 1.62), which lost its statistical significance by further adjustments. Stratified analysis indicated that metabolic dysfunction contributed to mortality (HR, 1.51; 95% CI, 1.21 to 1.89) and CVD (HR, 1.27; 95% CI, 1.02 to 1.59). Among metabolic dysfunctions used for defining MAFLD, type 2 diabetes mellitus in MAFLD increased the risk of both mortality (HR, 2.07; 95% CI, 1.52 to 2.81) and CVD (HR, 1.42; 95% CI, 1.09 to 1.85).ConclusionsMAFLD independently increased overall mortality. Heterogeneity in mortality and CVD risk of subjects with MAFLD may be determined by the accompanying metabolic dysfunctions.     

成人活体肝移植单中心56例临床报告

目的探讨成人间右半肝移植手术中保证供、受者安全的方法。方法2002年1月至2006年9月四川大学华西医院对56例受者施行了成人右半肝移植,其中52例不含肝中静脉,4例双供肝肝移植。受者原发病为乙型肝炎肝硬化35例（62．5％,含急性肝功能衰竭12例）,肝细胞肝癌17例（30．4％）,其他4例;MELD评分〉25分者10例。供者常规行三维CT计算全肝体积及右半肝体积,并进行了移植手术技术改进。结果58例供者共摘取55例右半供肝及3例左半供肝。右半供肝均不含肝中静脉,重量为400—860g（中位数550g）,右半供肝与受者标准肝重比为31．7％-71．7％（中位数45．4％）,供者残肝体积均大于全肝体积的35％。58例供者发生并发症7例（12．5％）,无死亡。术后住院时间7—30d（中位数11d）。术后对56例受者随访2—52个月（中位数11个月）,发生并发症15例（26．8％）,死亡4例（7．2％）。1年实际生存率92．8％。结论采用不包含肝中静脉的右半供肝,术前CT测量残肝体积〉35％,同时右半供肝与受者标准肝重比〉40％者进行右半肝移植可保证供受者安全,反之则应考虑采用双供肝肝移植。     

乳腺癌前哨淋巴结活检中亚甲蓝与亚甲蓝、同位素联合示踪的比较研究

目的比较单用亚甲蓝或联合亚甲蓝及同位素示踪剂在乳腺癌前哨淋巴结活检中的价值及研究影响前哨淋巴结活检准确性的因素。方法收集2005年3月至2006年7月期间行前哨淋巴结活检的164例乳腺癌患者,单用亚甲蓝示踪剂组104例,活检前乳晕旁皮下注射亚甲蓝2ml;亚甲蓝和同位素联合示踪剂组60例,术前肿瘤周围实质中4点注射^99m锝-硫胶体2mCi/4ml,经淋巴显像,术中在γ探测仪引导下行腋窝前哨淋巴结活检。结果前哨淋巴结总的检出率为97.0%（159/164）,准确性为95.0%（151/159）,假阴性率为14.0%（8/57）;联合组检出率、准确率、敏感性、阴性预测值高于亚甲蓝组,联合组的假阴性率为8.7%（2/23）,较亚甲蓝组17.7%的假阴性率为低（6/34）,但差异均无统计学意义（P=0.453）。亚甲蓝组平均每例检测出2.2枚前哨淋巴结,联合组为平均2.5枚（P=0.223）。前哨淋巴结数目≥3和〈3枚的患者假阴性率分别为0%和24.2%（8/33）（P=0.016）。联合组中发现73枚无蓝染仅有热点的前哨淋巴结,故同位素的应用将本组患者的检出率从85.0%（51/60）提高至98.3%（59/60）,假阴性率从21.7%（5/23）降低至8.7% （2/23）。结论在取得一定经验的情况下,亚甲蓝示踪与联合染料、同位素示踪可获得相似的前哨淋巴结检出率、准确率;同位素示踪剂的应用对亚甲蓝示踪起到很好的补充作用;前哨淋巴结检出越多,假阴性率越低。