骨肉瘤细胞特异性结合短肽的筛选

目的：获得与骨肉瘤细胞株os-732特异结合的短肽，作为骨肉瘤靶向治疗的先导化合物。方法：以骨肉瘤细胞os-732为靶细胞，成骨细胞为吸附细胞对噬菌体12肽库进行差减筛选，用细胞ELISA、免疫组化鉴定阳性噬菌体克隆并测序。结果：经三轮筛选，从随机挑选的20个噬菌体克隆中得到9个特异性与骨肉瘤细胞os-732结合，而不与正常成骨细胞结合的阳性克隆。但其氨基酸序列无同源性。结论：得到多个序列不同的特异性结合骨肉瘤的噬菌体克隆，提示骨肉瘤细胞表面结构复杂，具多个骨肉瘤抗原表位。本实验获得的短肽具有一定的亲合力和肿瘤特异性，为针对不同位点的靶向药物设计提供了实验依据。     

严重烧伤后肠道损伤与肠源性高代谢的关系

目的　探讨烧伤后肠道损害与“肠源性高代谢”的关系。方法　在建立严重烧伤加清洁肠道动物模型的基础上 ,将 96只Wistar大鼠随机分为烧伤对照 (B)组和烧伤加清洁肠道 (SDD)组。观察了伤后 0～ 10d大鼠静息能量代谢率(REE)的变化 ,同时检测了伤后 0、1、3、5、7、10d血中内毒素 (LPS)、肿瘤坏死因子 (TNF)和二胺氧化酶 (DAO)的含量 ,并进行相关分析。结果　烧伤后两组大鼠的REE、TNF、LPS和DAO均明显高于伤前 ,两组相比 ,SDD组的REE、TNF和LPS较B组均有不同程度的降低 ,而DAO则明显高于B组。相关分析显示 ,REE同LPS和TNF呈显著正相关 (r1 =0 .77,P <0 .0 5 ,r2 =0 .81,P <0 .0 5 ) ,与DAO相关不显著 (r >0 .0 5 )。结论　烧伤后肠源性高代谢与血中炎症介质含量呈正比 ,但肠道损伤程度加重并不引起代谢率相应增加 ,二者不存在线性关系     

热疗联合化疗与单纯化疗治疗非小细胞肺癌的疗效比较

目的：探讨热疗联合化疗治疗非小细胞肺癌（non—small cell lung carcinoma，NSCLC）的疗效。方法：2003年9月～2006年11月我院对52例NSCLC进行微波热疗联合NP方案化疗联合治疗（热疗联合化疗组），3个周期后评价疗效；并与2006年1～11月16例NSCLC单纯化疗（单纯化疗组）进行对比。结果：热疗联合化疗组近期疗效明显好于单纯化疗组，热疗联合化疗组总有效率（RR）为65．4％（34／52），显著高于单纯化疗组总有效率37．5％（6／16）（Z=-2．419．P=0．016）；热疗联合化疗组结束后疼痛明显改善，单纯化疗组无镇痛作用，差异具有高度统计意义（Z=-6．486，P=0．000）；2组的不良反应主要表现为恶心、粒细胞减少、血小板降低和肝功能异常，差异无显著性（P〉0．05）。结论：热疗联合化疗治疗NSCLC疗效优于单纯化疗治疗NSCLC。     

改良气囊导尿管内固定方法的临床探讨

目的探讨改良气囊导尿管的内固定方法，减轻患者的不适感，增加尿管留置成功率。方法将150例需留置导尿管的患者随机分为A、B、C3组，常规导尿成功后，A组向气囊内注入生理盐水12ml；B组向气囊内注入空气12ml；C组向气囊内注入生理盐水和空气各6ml，于24h后采用自制问卷调查患者不舒适的程度，并在1周后统计置管的成功率。结果A组Ⅱ度以上不舒适38例，留置失败1例，失败率为2．0％；B组Ⅱ度以上不舒适5例，留置失败12例，失败率为24．0％；C组Ⅱ度以上不舒适5例，留置失败2例，失败率为3．9％。满意度B组与C组比较差异无统计学意义，A组与B组和C组比较差并均有统计学意义（P〈0．01）。失败率A组与C组比较差异无统计学意义（P=0．3864），B组与A组和C组比较差异有统计学意义（P〈0．01）。结论3组从舒适的角度和留置成功率全面比较C组的方法优于A、B两组，留置气囊导尿管时向气囊内注入生理盐水和空气各6ml是内固定最佳的方法，在临床上是可行的。     

骨髓基质干细胞在体外向软骨细胞分化

目的观察骨髓基质干细胞(B M SC s)在体外能否分化为软骨细胞。方法利用高密度细胞球培养体系在含转化生长因子-β1(TG F-β1)的培养基中培养B M SC s21d,用免疫组化甲苯胺蓝染色方法分析培养的B M SC s球中蛋白多糖(软骨细胞分泌的主要基质成份)的表达、用免疫组化和R T-P C R方法分析Ⅱ型胶原(软骨细胞特异分泌的主要胶原蛋白)的表达来评估B M SC s是否分化为软骨细胞。结果TG F-β1作用的B M SC s表达了Ⅱ型胶原和蛋白多糖。结论B M SC s在体外特定的培养条件下可分化为软骨细胞,从而可能成为临床上治疗创伤或骨关节炎所致的软骨缺损所需的合适的自体来源的种子细胞。     

分叉带锁髓内钉治疗肱骨干骨折临床疗效分析

目的：探讨分叉带锁髓内钉治疗肱骨干骨折临床经验。方法：采用顺行和逆行分叉肱骨带锁髓内钉治疗肱骨干骨折25例。结果：随访平均26个月，优16例，良7例，可2例，优良率92％。除肩关节活动受限2例外，其余均完全恢复肩关节和肘关节功能。结论：分叉带锁髓内钉适应于肱骨骨折，其坚强－动态固定的有机结合有利于骨折的愈合。方法简单，固定可靠，创伤小，是一种较好的手术方法。     

睾丸网注射法建立转基因小鼠

目的 为了研究睾丸网注射法建立转基因小鼠的可行性。方法 本试验应用显微注射仪将脂质体包裹的质粒pEGFP—N1通过睾丸网注射到4周龄昆明（KM）小白鼠睾丸生精小管内。注射后6周，与自然发情雌鼠交配，用PCR法检测仔鼠基因组中的外源DNA。结果 共注射4只雄性小鼠，3只存活，并且具有交配，受精能力，共获仔鼠63只，其中2只为PCR阳性仔鼠。结论 用睾丸网注射法制备转基因动物是可行的。     

中药生精冲剂对精索静脉曲张大鼠的治疗

目的：研究中药生精冲剂对大鼠精索静脉曲张的影响及疗效。方法：从80只SD雄性大鼠中随机抽出20只作为假手术组，余60只均建立精索静脉曲张病理模型后随机均分为模型组、生精冲剂组和克罗米芬组。造模后15d生精冲剂组和克罗米酚组分别给予生精冲剂4g/（kg·d）和克罗米芬20mg/（kg·d）灌胃，模型组和假手术组正常喂食。造模后45d放免法测定血清性激素（FSH、LH和T）及观察各组大鼠睾丸组织结构。结果：生精冲剂组大鼠光镜下睾丸组织结构优于模型组和克罗米芬组；血清FSH、LH生精冲剂组显著低于模型组和克罗米芬组（P〈0.05），而克罗米芬组显著高于其他3组。T在生精冲剂组、克罗米芬组和假手术组之间无显著差异，但均显著高于模型组（P〈0.05）。结论：中药生精冲剂对精索静脉曲张引起的睾丸损害有保护及修复作用，且可能优于克罗米芬。     

Effect of curcumin on nitric oxide synthase expression in Iipopolysaccharide-activated microglia cells and the anti-oxidative effect of curcumin

BACKGROUND: It has been demonstrated that curcumin can increase the activities of various anti-oxidase in blood and tissue, effectively eliminate various free radicals, reduce the production of peroxisome, and alleviate oxidative stress reaction. Whether it has the same effect on microglia? OBJECTIVE: To observe the effects of curcumin on the expressions of inducible nitric oxide synthase (iNOS), nuclear factor-κB (NF-κB), and superoxide dismutase (SOD) in microglial cell line BV stimulated by lipopolysaccharide (LPS). DESIGN: An observational comparative study. SETTING: Research Room of Biochemistry, Medical College of Nantong University. MATERIALS: Mice microglia cell line BV, iNOS and NF-κB reporter gene plasmids were presented by Dr. Bhat.NR. from the Medical University of South Carolina (USA). Curcumin was produced by the Xi'an Branch of China Chengdu Scholar Bio-Tech. Co.,Ltd.; LPS (E.Coli O26:B6), anti-mice iNOS monoclonal antibody, horseradish peroxidase labeled goat-anti-mice IgG were the products of Sigma Company (USA). METHODS: The experiments were carried out in the Research Room of Biochemistry, Medical College of Nantong University from May 2006 to April 2007. ① Detection of iNOS: The cells were seeded onto 24-well plate at the density of 1×105, After the cells had adhered to the cover glasses, the cells were grouped as negative control group (the primary antibody was replaced by phosphate buffered solution PBS); normal control group (the cells were normally cultured); LPS-treated group (the cells were treated with LPS for 24 hours); curcumin+LPS group (the cells were treated with curcumin for 1 hour and LPS for 24 hours). The expressions of iNOS protein were detected with immunocytochemical staining. ② Determination of iNOS and NF-κB gene activities: According to the introduction of the kit for transfection, iNOS or NF-κB report gene plasmids were transiently transfected with LipofectamineTM2000 liposomes into the cells in the 24-well plate for 24 hours. The cells were divided into normal control group (the cells were normally cultured after transfected with report gene plasmids); blank plasmid group (the cells were normally cultured after transfected with blank plasmids); LPS-treated group (the cells were treated with LPS for 4 hours after transfected with report gene plasmids); curcumin+LPS group (the cells were treated with curcumin for 1 hour and LPS for 24 hours after transfected with report gene plasmids). The content of luciferase in the cell lysis buffer was determined after cell lysis. ③ Determination of SOD activity: The cells were seeded into culture bottle at the density of 1×106, and the divided into four groups, including normal control group (the cells were normally cultured); LPS-treated group (the cells were treated with LPS for 24 hours); curcumin+LPS group (the cells were treated with curcumin for 1 hour and LPS for 24 hours); vitamin C+LPS group (the cells were treated with vitamin C for 1 hour and LPS for 24 hours). The SOD activity was determined with xanthine oxidase and quantitative colorimetric assay. MAIN OUTCOME MEASURES: The expressions of iNOS protein, iNOS and NF-κB, and the activity of SOD were observed. RESULTS: ① Expression of iNOS protein in microglia: The expression of iNOS protein in the LPS-treated group was obviously higher than that in the negative control group (P < 0.01); Those in the curcumin+LPS group were significantly decreased as compared with that in the LPS-treated group (P < 0.01). ② Expressions of iNOS and NF-κB genes: The expressions of iNOS and NF-κB genes in the LPS-treated group were significantly higher than those in the normal control group (P < 0.01); Those in the curcumin+LPS group were significantly lower than those in the LPS-treated group (P < 0.01). ③ SOD activity: The activity of SOD in the LPS-treated group was significantly lower than those in the normal control group (P < 0.01). It in the curcumin+LPS group and vitamin C +LPS group was significantly higher than that in the LPS-treated group (P < 0.01). CONCLUSION: Curcumin could inhibit the expression of iNOS in the activated microglia, and it also has the abilities in eliminating free radicals and antagonizing lipid peroxidation.