1198例非体外循环冠状动脉旁路移植术的早期临床分析

目的　总结分析非体外循环冠状动脉旁路移植术 (OPCAB)的早期临床结果和经验体会。方法　 1996年 10月至 2 0 0 2年 5月在国内 15个冠心病微创外科中心完成非体外循环冠状动脉旁路移植术 1198例 ,占同期冠心病手术的 79 5 % ,OPCAB实施率 (即OPCAB占单纯冠状动脉旁路移植术的百分比 )为 85 2 %。年龄平均 (6 1 6± 9 0 )岁 ,男性占 77 3%。 18 7%为左主干病变 ,76 9%为 3支病变。左心室射血分数平均 0 5 6± 0 17。其中二次手术者占 1 5 % ,急诊手术占 9 9%。结果　 2 2例为小切口单支病变旁路移植手术 ,其余均为正中切口、多支病变的OPCAB手术。远端吻合口为 (3 1± 0 9)个 ,乳内动脉桥占 6 5 6 % ,桡动脉桥占 18 0 % ;5 2 %病人使用主动脉内球囊反搏。住院死亡 14例 ,病死率 1 2 %。其中术后心跳骤停或室颤 7例 ,大出血 2例 ,昏迷伴肾功能衰竭 2例 ,大面积脑梗塞 2例 ,呼吸衰竭 1例。并发症中 ,围术期心肌梗死占 0 4% ,急性左心功能衰竭占 0 3% ,严重心律失常占 1 2 % ,呼吸系统并发症占 2 0 % ,器质性神经系统并发症占 1 2 % ,新发或加重的肾功能不全占 0 7% ,出血二次开胸占1 0 % ,胸骨愈合不良占 0 6 %。结论　OPCAB手术安全可行 ,早期效果满意 ,远期效果有待进一步观察。OPCAB具备一     

不同浓度5-溴脱氧尿嘧啶核苷标记山羊骨髓基质干细胞的体外研究

Objective To explore the feasibility of labeling and tracing in vitro goat bone marrow mesenchymal stem cells (BMSCs) by bromodeoxyuridine (BrdU) on the basis of investigation of its optimal concentration, incubating time and cytotoxicity. Methods A healthy goat, aged 10 months old, male, weighing 32 kg, was used in this study. Bone marrow was aspirated. BMSCs were isolated and cultured using the adherence method in vitro. The fourth passage of BMSCs (P4) were incubated with BrdU at 5, 10, 15, 20 μmol/L as 5, 10, 15, 20 μmol/L BMSC groups. Cells were not labeled by BrdU as negative control. The following parameters were measured: induction, differentiation and determination of goat BMSCs; the optimal mass concentration and incubation time of 5-BrdU labeling; cell positive rate at 12, 24, 48 and 72 hours in each group using immunofluoreseenee; the cell survival rate after various concentrations of BrdU ladling by trypan-blue exclusion. Results The morphology of the primary and passage goat BMSCs was fusiform in shape. Goat BMSCs could differentiate into osteoblasts and chondrocytes following induction. BMSC nucleus showed green fluorescence under fluorescence microscope after being labeled by BrdU. The mean labeling rate increased with the increase in the concentration and incubation time of BrdU, and reached to (93.32± 3.25)% after incubation in 15 μmol/L, BrdU for 48 hours. There were no significant differences between 15 μmol/L BrdU for 72 hours, 20 μmol/L BrdU for 48 hours and 72 hours (P > 0.05), or between the other groups or time points (P < 0.05). The labeling rate of the blank control group was 0. The cell survival rate was all above 90% (P > 0.05). Conclusions BrdU can be used as a labeling marker for goat BMSCs. When the concentration is 15 μmol/L and the incubation time is 48 hours, the optimal labeling effect can be achieved. Goat BMSCs labeled with BrdU is of high efficiency and safety.     

基于慢病毒的SHARP-2特异的RNA干扰延长肾移植受者大鼠存活时间

Objective To investigate if rat enhancer of split- and hairy-related protein-2 (SHARP-2) short hairpin RNA interference (shRNAi) prolongs the survival time of rat kidney transplant recipients. Methods Gene recombinant procedures, transfection and co-transfection were carried out to introduce short hairpin RNA interference sequences target for SHARP-2 into 3rd generation self-inactivated lentiviral-ViraPower packaging mix. Limiting dilution method was used for viral titration. Real-time PCR was employed for quantification of gene expression. Rat kidney transplantation was utilized to investigate the effect of SHARP-2 gene silence on the recipient survival. Results A lentiviral-based shRNAi construct LV-SHARP-2iC showed 84% SHARP-2 gene silence efficiency in normal rat kidney cells. At multiplicity of infection 20, 57% T cells could be transfected by lentivirus with spinoculation method. In activated T cells, SHARP-2 g ene silence resulted in 61.3% and 68.7% reduction of intedeukin 2 (IL-2) and interferon γ (IFN-γ) gene expression. When donor kidney was perfused with 5×107 TU LV-SHARP-2iC, the median survival time prolonged for 4-5 days as compared to blank and scramble control groups. Conclusions A recombinant lentivirus LV-SHARP-2iC that effectively silence SHARP-2 gene expression is constructed successfully, leading to the inhibition of IL-2 and IFN-γ. LV-SHARP-2iC treatment can prolong the survival time of rat kidney transplant recipients.     

Novel multi-probe RNase protection assay set for detection of endotoxin associated receptors gene expression

Objective: To construct the multi-probe ribonuclease protection assay (RPA) template set to be used for detecting expression patterns of MD-2, TLR4, CD14 mRNAs in human peripheral blood mononuclear cells. Methods : The designed cDNA fragments of the three genes were generated by polymerase chain reaction (PCR)using specific primers and directionally cloned into EcoR I and Hind III sites of expression plasmid pSP72 containing the T7/ promoter, the linearized plasmids was used as template to synthesize anti-sense RNA probes. Then we extracted total RNA from peripheral blood mononuclear cells (PBMC) and detected the dynamic expression patterns of the three genes with RPA method. Results: The proper sequence and orientation of the template set were confirmed by sequencing and the template set was successfully used to assay TLR4, MD-2 and CD14 mRNAs in human PBMC. The results showed that the three detected genes decreased transiently 1-3 hours after 100 ng/ml LPS stimulation. Conclusions: These new RPA multi-probe set provided valuable tool for the simultaneous quantitative determination of expression of TLR4, CD14 and MD-2 mRNAs in both constitutive and inducible types.     

碳增强的聚醚醚酮作为髋臼假体材料的实验研究

目的:评价碳纤维增强的聚醚醚酮(CFPEEK)作为髋臼假体材料的生物相容性和耐磨损性。方法:(1)采用国产材料和工艺研制出碳纤CFPEEK复合材料,在模拟体液环境下对其本身以及与不同磨擦偶的耐磨损性能进行测试,并与传统髋臼假体材料超高分子聚乙烯(UHMWPE)作比较。(2)体外研究CFPEEK磨屑对巨噬细胞ANA-1释放白细胞介素1(IL-1_β)和肿瘤坏死因子α(TNF-α)的影响,并与传统的人工关节材料比较,以确定各材料间生物学反应的差异。结果:(1)CFPEEK的体积磨损率只为UHMWPE的1/2,耐磨损性能明显优于UHMWPE(Alpha=0.01水平)。CFPEEK与钴铬钼合金(CoCrMo)对磨将产生最少的磨屑(Alpha=0.01水平)。CFPEEK与CoCrMo对磨时的摩擦系数最低。(2)各实验组IL_(-1β)和TNF-α的释放都有增高,实验结果总体差异有显著性意义(P=0.0001),UHMWPE组明显高于CFPEEK组(Alpha=0.01水平)。结论:CFPEEK材料具耐磨损、生物相容性好的特点,是一种未来髓臼假体的理想材料。     

二氢青蒿素对胰腺癌生长的抑制作用

Objective To investigate the anti-tumor activity of dihydroartemisinin in pancreatic cancer in vitro and in vivo. Methods For cultured cells,cell growth was determined by the MTT assay and apoptosis was evaluated by flow cytometry analysis stained with Annexin V-FITG/PI. The protein expression in BxPC-3 cells was analyzed by Western blot assay. BxPC-3 cells were injected subcutaneously into nude mice to establish pancreatic xenograft tumors and the tumor volume was monitored after exposure to dihydroartemisinin. Ki-67 staining and TUNEL assay were used to assess tumor cell proliferation and apoptosis in tumor tissue. Results After treatment by dihydroartemisinin in vitro, the proliferative inhibition rates of pancreatic cancer cells BxPC-3 and AsPC-1 reached up to (76.2 ± 3.5) % and (79.5 ± 2.9) %, and the apoptosis rates were up to (55.5 ± 3.2)% and (40.0 ± 3.5)%, the differences were significantly (P ＜ 0.01) compared with control [(2.0 ± 1.3) % and (0.9 ± 0.4) %]. Dihydroartemisinin inhibited the growth of pancreatic xenograft tumors in nude mice. The proliferation index and apoptusis index were (49.1 ± 3.9)% and (50.2 ± 4.4)% respectively in dihydroarternisinin 50 mg/kg treatment group, compared to those of (72.1 ± 3.3) % and (9.4 ± 2.9) % in control, the differences were significantly (P ＜0.01). Western blot assay indicated that dihydroartemisinin up-regulates expression of proliferation-associated protein p21WAF1 and down-regulates expression of PCNA, increases expression of apoptosis-associated protein Bax and decreases expression of Bcl-2 and activates caspase-9 in BxPC-3 cells. Conclusions Dihydroartemisinin exerts anti-tumor activity in pancreatic cancer both in vitro and in vivo by proliferation inhibition and apoptusis induction. Dihydroartemisinin can be used as a potential anti-tumor drug in pancreatic cancer.     

踝关节骨折的手术治疗

目的 探讨手术治疗踝关节骨折的疗效.方法 回顾性分析2000年1月至2008年4月手术治疗且有完整随访资料的92例踝父节骨折患者的疗效.评价指标包括美国足踝机会踝与后足功能评分(AOFAS评分)、骨折部位的疼痛评估(VAS评分)、患者满意度、踝关节活动度.并对踝关节健侧与患侧的关节活动度差异,以及不同AO分型之间、骨折累及不同部位(单踝、双踝、三踝骨折)之间、不同随访时限之间这二种情况下的踝关节屈伸活动度及AOFAS评分的差异进行统计学分析.结果 平均随访55个月,所有患者骨折均愈合;AOFAS评分平均为93.6分,优良率100%;VAS评分平均为0.17分,患者满意度评分平均为9.62分.健、患侧踝关节活动度差异有统计学意义(P<0.05),不同AO分型之间、骨折累及小同部位之问、不同随访时限之问的踝关节屈伸活动度和AOFAS评分差异无统计学意义(P>0.05).结论 手术治疗踝关节骨折可取得满意的临床疗效.     

FK506-壳聚糖膜与体外培养雪旺细胞生物相容性的研究

目的 探讨FK506-壳聚糖膜片与培养的雪旺细胞生物相容性.方法 将生长状况和纯度较好的第3代雪旺细胞,分别传代接种于含有FK506-壳聚糖膜片、单纯壳聚糖膜片的培养皿中培养,以盖玻片作为对照组.计算3组雪旺细胞倍增时间、雪旺细胞纯度;用MTT法比较不同膜片上雪旺细胞的生长活力,绘制生长曲线;S-100免疫荧光染色.结果 雪旺细胞倍增时间对照组为5.9 d,单纯壳聚糖膜片及FK506-壳聚糖膜片组均为4.0 d;雪旺细胞纯度分别为80%、89%,93%;雪旺细胞增殖情况以FK506-壳聚糖组最佳,单纯壳聚糖组次之,对照组最差;各组雪旺细胞均呈S-100阳性,FK506-壳聚糖组与单纯壳聚糖组雪旺细胞排列呈典型的漩涡状或栅栏状.结论 FK506-壳聚糖膜片具有更显著的组织相容性与生物功能性,并保持了雪旺细胞的生物学特性.     

谷氨酰胺和生长激素对门静脉高压症患者术后营养、免疫和炎症反应的影响

Objective This study was conducted to evaluate the effects of postoperative glutamine and growth hormone on the nutritional status,immune function,and inflammatory responses of patients undergo-ing portal hypertension surgery.Methods This study was designed as a prospective,randomized and con-trolled clinical trial.Forty-two patients undergoing portal hypertension surgery were randomly divided into 2 groups:experimental group (receiving an immune enhancing parenteral nutrition by adding glutamine and re-combinant human growth hormone,n = 22),and control group (receiving an isocaloric and isonitrogenons standard parenteral nutrition,n = 20).Parenteral nutrition providing 125 kJ · kg-1·d-1 was initiated 3 days after surgery in both groups and last for 7 days.Blood samples were obtained on day 0th,3rd,and 10th.Host nutritional status was evaluated by measuring levels of prealbumin and transferrin,immunity was evalu-ated by measuring levels of CD4,CD8,CD4/CD8,IgG,IgM and IgA,and the inflanunatory responses was determined by assessing IL-2,TNF-α and C-reactive protein (CRP) levels.Results Ten days after opera-tion,among patients receiving an immune enhancing parenteral nutrition,the serum prealbumin was (193.84±39.78)mg/L,transferrin was (2.07±0.51)mg/L,CD4 was (33.7±5.5)%,CD4/CD8 was (1.17±0.32),IgG was (13.94±1.09)g/L and IL-2 was (368.12±59.25) pg/mL.They were signifi-cantly higher than those of the control group (P < 0.05).The TNF-α concentrations was (321.12±81.42)pg/mL and CRP was (32.2±15.2)mg/mL which were significantly lower than those of the control group (P < 0.05).Conclusion Postoperative administration of immune enhancing parentearl nutrition in patients undergoing portal hypertension surgery can improve nutritional status and immune function,and modulate inflammatory response is also better than standard one.     

中央型颈脊髓损伤保守与手术治疗的对比研究

目的对比研究手术与非手术治疗中央型颈脊髓损伤的临床疗效。方法中央型颈脊髓损伤患者120例,手术治疗72例,其中前路手术28例,后路手术44例;非手术治疗48例。详细记录患者ASIA运动功能评分(AMS),对治疗前及治疗后6个月、2年时的AMS评分进行比较。结果96例获随访,时间6个月~5年,21例失访,3例死亡。两组治疗后6个月、2年时AMS评分之间差异有显著性(P<0.05)。治疗后6个月内非手术组6例出现并发症(6/48,12.5%),手术组4例出现并发症(4/72,5.6%),两组之间差异无显著性(P>0.05)。结论手术治疗中央型颈脊髓损伤疗效可靠。早期手术能够解除脊髓压迫,稳定颈椎,促进脊髓功能恢复。