Buccal mucosal graft for urethral reconstruction.

Urethral reconstruction with graft substances, such as skin and bladder mucosa, has been previously used when primary anastomosis cannot be achieved. However, stricture and meatal prolapse are associated with these grafts. We report the use of buccal mucosa for the reconstruction of urethral defects in 3 patients. One patient with failed operation for hypospadias received tube buccal mucosal graft for urethral replacement. Two patients with urethral necrosis and stricture received onlay buccal mucosal graft. All patients were disease-free during follow-up (range, 12 to 49 months; mean, 36 months). One patient had a pinhole fistula that was successfully managed with simple repair. This technique appears to be useful for urethral reconstruction when a local graft is not available, even in patients with complicated conditions.     

The characteristics of Hep-2 cell with multiple drug resistance induced by Taxol.

OBJECTIVE: To investigate the characteristics of Hep-2 cell with multidrug resistance (MDR) induced by Taxol. STUDY DESIGN: Hep-2 cells were exposed in stepwise escalating concentration of Taxol to develop the resistant cell line-Hep-2T. Cell cycle distribution, apoptosis, and rhodamine accumulation were studied through flow cytometry. The MDR1 and MRP1 genes were detected through real-time quantitative RT-PCR, and the corresponding proteins were detected through Western blotting. RESULTS: The drug resistance of Hep-2T cells to Taxol, doxorubicin, gemcitabine, 5-FU, and cisplatin all increased. The percentage of G0/G1 phase and the antiapoptosis ability increased significantly compared with Hep-2 cells. Both MDR1 and MRP1 also increased at gene and protein level, though MDR1 was more prominent. CONCLUSION: More emphasis should be laid on MDR1/Pgp, the non-Pgp substrate chemotherapeutic agents, and the changes of cell cycle distribution to prevent MDR induced by Taxol. SIGNIFICANCE: These findings may provide theoretical support for the reverse of MDR.     

Measurement of γ-enolase release, a new method for selective quantification of neurotoxicity independently from glial lysis

We have developed a sensitive enzymatic-immunoassay to quantify the level of gamma-enolase (a specific neuronal enzyme) which is released from cultured cells after exposure to various toxins. We show that this method can estimate selectively neuronal cell death without significantly interfering with glial cell death. Indeed, no gamma-enolase is released when glial cells are killed with free-radical producing agents. Experiments comparing the levels of neuronal cell death induced by NMDA or free-radical producing drugs, performed either by measuring gamma-enolase release or using the classical fluorescein diacetate method, yielded similar results. In addition to selectively follow neuronal death in a mixed population of neurons and glial cells, this method provides a way of determining the cell death kinetics from a single culture dish, since enolase can be measured on small samples taken from the culture medium. Finally, we propose these two methods as being complementary and useful neuronal and other cellular death indexes and also to understand the complex problem of glial influence on neuronal survival or death.     

永瓣藤化学成分的研究

永瓣藤化学成分的研究陈玲，谢平，戚薇，曾菊珍（药学系中药化学教研室）关键词永瓣藤，化学成分，三萜，蒜盘子甾酮永瓣藤（ＭｏｎｉｍｏｐｅｔａｌｎｍＣｈｉｎｅｎｓｅＲｅ－ｈｄ）系卫矛科永瓣藤属植物［１］，为我国特有的珍稀二类保护植物［２］，分布于江西东北部...     

视网膜特异性表达人血管内皮生长因子 基因载体的构建

目的构建在视网膜组织特异性表达的人血管内皮生长因子（VEGF）₁₆₅基因。方法用聚合酶链反应(PCR)方法从BLAB/C鼠全基因组扩增能在视网膜组织特异性表达的rho启动子，经限制性内切酶纯化后克隆于质粒pcDNA^3.1+-VEGF₁₆₅中，建立重组质粒pcDNA^3.1+-rho-VEGF₁₆₅，通过限制性内切酶酶切分析及PCR鉴定筛选出正确重组质粒pcDNA^3.1+-rho-VEGF₁₆₅,由jetPEI介导转染人视网膜色素上皮细胞和人脐静脉内皮细胞，并通过免疫组织化学染色以及绘制细胞生长曲线检测在人视网膜色素上皮细胞和人脐静脉内皮细胞中VEGF蛋白的表达。结果在人视网膜色素上皮细胞中，重组质粒pcDNA^3.1+-rho-VEGF₁₆₅比质粒pcDNA^3.1+-rho-VEGF₁₆₅的VEGF蛋白表达强，在人脐静脉内皮细胞，两者的表达量无明显差别。结论pcDNA^3.1+-rho-VEGF₁₆₅载体的构建为进一步研究VEGF在视网膜新生血管形成中的致病机理提供基础材料，并为进一步建立视网膜特异性表达VEGF转基因鼠模型建立了基础。(中华眼底病杂志,2005,21:106-109)     

Management of chronic lunotriquetral ligament tears

Treatment of chronic disruptions of the lunotriquetral (LT) ligament is not well-defined. Eleven patients treated by LT fusion with use of a compression screw are reported. The injury frequently resulted from hyperextension of the wrist. Pain on the ulnar side of the wrist, limited motion, and tenderness over the LT joint exacerbated by ballottement were present. Standard radiographs were normal. Arthrography showed the ligamentous tear in all cases. After operation, immobilization was continued until fusion was apparent radiographically. Fusion was achieved in all cases between 2 and 5 months. Four patients were free of pain, four patients had pain only at the extremes of motion, and three patients had persistent pain. Mean wrist motion was as follows (preoperative/postoperative): flexion (53 degrees/45 degrees), extension (60 degrees/49 degrees), radial deviation (17 degrees/21 degrees), and ulnar deviation (25 degrees/18 degrees). Maximum grip strength as a percentage of the uninjured side was 73% preoperatively and 59% postoperatively. LT tears can exist de novo or as part of the ulnar impaction syndrome; a method for differentiation is presented.     

Long-term expression of the c-fos protein during the in vitro differentiation of cerebellar granule cells induced by potassium or NMDA.

Levels of the c-fos protein were assayed in mouse cerebellar granule cells during their in vitro development under different culture conditions. When grown in media favoring both their survival and differentiation, i.e. in the presence of 30 mM K+ or 12.5 mM K+ plus 100 microM N-methyl-D-aspartate (NMDA), the c-fos protein becomes detectable in the nucleus of granule cells on and after 6 days and persists to high levels until the culture begins to decline. The protein c-fos appears therefore after the critical period described for the survival effect of K+ depolarization or NMDA receptor stimulation which corresponds to days 2-5 after plating. The c-fos protein remains however scarcely detectable or undetectable throughout the life-span of cells cultured under conditions providing poor survival and differentiation, i.e. in the presence of low K+ (5 or 12.5 mM) alone or when the effect of NMDA is blocked by the NMDA receptor antagonist MK-801. Interestingly, in cortical and striatal neurons, the survival and differentiation of which being not affected by depolarizing media, no c-fos protein is detected whatever the culture conditions tested at least during the first 18 days in vitro. This suggests that long-term expression of the c-fos gene might be related to some aspect of the late in vitro differentiation process of cerebellar granule cells.     

脉动真空灭菌器内室气体流向监测报警装置的研制

本文针对XGI型脉动真空灭菌柜出现的蒸汽倒流故障设计出一种内室气体流向监测报警装置，对管道中的蒸汽流向和空气流向进行监测。当出现倒流现象时，报警装置发出报警信号，提醒工作人员采取措施排除故障。经在两台灭菌柜上使用，证明此方案是一种行之有效的方法。     

In vitro synthesis of antimycobacterial antibodies with different specificities in various tissues of leprosy patients

For the detection of the synthesis in vitro of anti-Mycobacterium leprae antibodies in various tissues of leprosy patients, biopsy specimens of skin lesions, nasal mucosa, larynx, lymph nodes, and bone marrow were cultured in a medium containing 14C-labeled lysine and isoleucine. The culture fluids were analyzed by crossed immunoelectrophoresis with intermediate gel and autoradiography. The results show that synthesis of anti-M. leprae antibodies occurs at the investigated sites of leprosy patients and that the specificities of the synthesized antibodies differ between sites in individual patients. It is conceivable that these antibodies play a role in the local defense against M. leprae.     

菠菜乙醇酸氧化酶编码区cDNA在酵母表达载体中的克隆

目的：克隆菠菜乙醇酸氧化酶cDNA并构建其酵母表达载体。方法：从新鲜菠菜叶子中提取总RNA，用RT－PCR法扩增菠菜乙醇酸氧化酶编码区cDNA并插入pMD-T载体中进行序列测定；将此cDNA构建入P.Pastoris酵母表达体系中的pPIC3.5k上的SnaBI/NotI位点，进行PCR筛选和限制性酶切鉴定。结果：测序表明获得的基因为乙醇酸氧化酶cDNA序列，与国外文献报道的序列相比有约98％的同源性；重组质粒的SnaBI/NotI双酶切证明构建正确。结论：菠菜乙醇酸氧化酶cDNA克隆及重组质粒pPIC3.5K-GO的获得，为酵母表达菠菜乙醇酸氧化酶提供了前题条件。