结肠癌细胞HIF-1α及MDR1 miRNA重组慢病毒干扰体系多药耐药研究

目的 研究HIF-1α及MDR1 miRNA重组慢病毒干扰体系的多药耐药性.方法 收集结肠癌切除的标本60例,用福尔马林(10％)浸泡,制成蜡块存档标本.通过观察HIF-1α与P-gp的表达和分布情况,找出二者关联.并构建miRAN干扰体系,研究重组慢病毒干扰体系和结肠癌细胞多药耐药的关系.结果 HIF-1α的表达与患者的性别、年龄、肿瘤的位置以及肿瘤分化的程度无关(p＞0.05),而与Dukes分期、淋巴结转移有关.HIF-tα表达与P-gp的表达呈显著正相关(γ=0.613,P＜0.05).LoVo细胞对ADR、VCR及5-Fu的敏感性均有不同程度的恢复,但沉默HIF-1α前后并无统计学差异(P＞0.05),沉默MDR1基因后,LoVo细胞对ADR与VCR的耐药性有所逆转,沉默MDR1基因表达对二者化疗敏感性变化无明显影响.结论 HIF-1α与P-gp的表达不受患者的性别、年龄、肿瘤位置、分化程度的影响,HIF-1α表达与P-gp的表达呈显著正相关,说明HIF-1α通过与MDR1/P-gp的相互作用最终参与了结肠癌多药耐药的发生.     

谷氨酰胺联合脐血间充质干细胞移植在大鼠肠缺血再灌注损伤中的作用

目的探讨谷氨酰胺联合脐血间充质干细胞(MSCs)移植在大鼠肠缺血再灌注损伤中作用。方法体外复苏并培养脐血间充质干细胞移植前备用,观察CM-Di I荧光标记后脐血间充质干细胞的去向。80只SD大鼠随机分为正常对照组,缺血再灌注损伤组,谷氨酰胺组,MSCs移植组及联合组每组各15只。对照组采用生理盐水灌肠,损伤组采用TNB(S乙醇稀释)灌肠,在TNBS建模后1 h,谷氨酰胺组于尾静脉输入谷氨酰胺0.45 g/kg、MSCs移植组于尾静脉输入1×10~(10)/L脐血间充质干细胞悬液,联合组尾静脉输入谷氨酰胺0.45 g/kg+脐血间充质干细胞悬液1×10~(10)/L。通过ELISA法检测各组大鼠血清中肠脂肪酸结合蛋白(IFABP)、白介素-6(IL-6)、超氧化物歧化酶(SOD)的含量;各组于再灌注1 h、3 h后检测肠组织含水率;通过RT-PCR、Western blot观察大鼠肠黏膜上皮细胞caspase-3、NF-k B、Bcl-2在谷氨酰胺联合MSCs移植后的mRNA和蛋白的表达情况。结果通过荧光示踪法观察到移植的MSCs细胞分布于肠粘膜淋巴组织内和腺上皮细胞间,表明MSCs可能参与了肠缺血再灌注损伤的修复过程。各组大鼠血清中SOD、IFABP、IL-6的含量变化比较,损伤组血清中IFABP、IL-6的含量较对照组显著增加,而谷氨酰胺组,MSCs移植组及联合组与之比较,则显著减少,联合组减少更为明显,损伤组血清中SOD的含量较对照组显著减少,而谷氨酰胺组,MSCs移植组及联合组与之比较,则显著增高,联合组增高更为明显(P0.05)。再灌注1 h和3 h,损伤组肠组织含水率均明显高于对照组;与损伤组相比,谷氨酰胺组、MSCs移植组及联合组肠组织含水率值均显著降低,联合组降低更为明显,而谷氨酰胺组、MSCs移植组差异无统计学意义(P0.05)。与对照组比较,损伤组肠黏膜上皮细胞caspase-3、NF-k B的mRNA和蛋白表达明显上调,Bcl-2的mRNA和蛋白表达明显下调(P0.05),而谷氨酰胺组、MSCs移植组及联合组与之比较,caspase-3、NF-k B的mRNA和蛋白表达明显下调,Bcl-2的mRNA和蛋白表达明显上调(P0.05),谷氨酰胺组及MSCs移植组之间无统计学差异(P0.05),但两组与联合组比较,差异明显(P0.05)。结论谷氨酰胺组及MSCs移植后,明显减轻了大鼠肠缺血再灌注损伤程度,其可能通过抑制caspase-3、NF-k B表达和促进Bcl-2表达减轻肠黏膜缺血再灌注损伤。     

脐血间充质干细胞移植修复急性肠缺血再灌注损伤

BACKGROUND:Bone marrow mesenchymal stem cells can repair intestinal ischemia-reperfusion injuny by interfering inflammatory reactions after intestinal ischemia-reperfusion to protect intestinal barrier functions. In recent years, umbilical cord blood mesenchymal stem cells are gradually used as a substitute source of bone marrow mesenchymal stem cells. OBJECTIVE:To investigate the effects of umbilical cord blood mesenchymal stem cells on acute intestinal ischemia-reperfusion injury. METHODS:Umbilical cord blood mesenchymal stem cells were induced, isolated in vitro and tracked by CM-DiI fluorescent labeling. Sixty-three Sprague-Dawley rats were equivalently randomized into three groups: control group received normal saline enema, intestinal ischemia-reperfusion injury group with ethanol diluted trinitro-benzene-sulfonic acid and transplantation group administrated with 1×10¹⁰/L umbilical cord blood mesenchymal stem cell suspension via the tail vein at 1 hour after trinitro-benzene-sulfonic acid modeling. At 3 days after transplantation, colon tissues were removed in each group to observe pathological changes of the intestinal tract by hematoxylin-eosin staining. Besides, expression of leptin mRNA in the colon tissues and cyclooxygenase-2 in the mucosa were detected by RT-PCR and immunohistochemistry method, respectively. RESULTS AND CONCLUSION:Transplanted umbilical cord blood mesenchymal stem cells distributed in the intestinal lymphoid tissue and among glandular epithelial cells, suggesting that these stem cells might be involved in the process of intestinal ischemia-reperfusion injury repair. Compared with the control group, intestinal injury in the injury group was significantly aggravated, and most intestinal epithelial cells shed; and the transplantation group appeared to have significantly reduced intestinal damage and significantly less cell shedding. Expression of leptin mRNA was significantly higher in the injury group than the transplantation group followed by the control group, and there were significant differences among the three groups (P < 0.05). Additionally, expression of cyclooxygenase-2 in the injury group was significantly higher than that in the control group (P < 0.05); compared with the injury group, expression of cyclooxygenase-2 was significantly lower in the transplantation group (P < 0.05). To conclude, leptin and cyclooxygenase-2 may be involved in acute intestinal ischemia-reperfusion injury, and umbilical cord blood mesenchymal stem cell transplantation significantly lessens intestinal ischemia-reperfusion injury, which provides an experimental basis for human treating acute intestinal ischemic injury.     

晚期先天性髋关节脱位的手术治疗与康复

目的：探讨晚期先天性髋脱位手术与康复疗效。方法： 采用高位髂骨截骨造盖术与康复训练等手段。结果：效果明显高于未进行者。结论：手术结合康复干预， 为目前治疗晚期先天性髋脱位较好的康复方法     

脑瘫患儿血浆白细胞介素6、肿瘤坏死因子α含量变化

有关脑瘫患儿的免疫功能,以往研究甚少[1]。我们对46例脑瘫患儿血浆白细胞介素6（IL－6）、肿瘤坏死因子-α（TNF－α）含量进行了检测,以探讨IL－6、TNF－α等免疫因素在脑性瘫痪发生发展中的作用机制。对象与方法一、对象脑瘫患儿46例．男30例．女16例,年龄1～10a。其中痉挛型22例,手足徐动型10例．混合型（痉挛型 手足徐动型）12例,共济失调型2例。运动障碍程度：轻度4例,中度22例,重度14例,极重度6例。脑性瘫痪的诊断、分型及运动障碍程度分类根据第互届全国小儿脑瘫座谈会标准。正常对照组24例,男14名,女10名,年龄1～1…     

晚期先天性髋关节脱位的手术治疗与康复

目的：探讨晚期先天性髓脱位手术与康复疗效。方法：采用高位髂骨截骨造盖术与康复训练等手段。结果：效果明显高于未进行。结论：手术结合康复干预，为目前治疗晚斯先天性髓脱位较好的康复方法。