用SSH方法构建BTCC患者尿脱落细胞差异表达基因的cDNA消减文库

目的:应用抑制性消减杂交(SSH)方法构建膀胱移行细胞癌(BTCC)患者与正常人尿脱落细胞差异表达基因cDNA消减文库。方法:分别从BTCC患者与正常人尿液中提取总mRNA,用SMART技术反转录成cDNA,经过HaeⅢ酶切后将BTCC尿脱落细胞cDNA分为两组并接上接头,再与过量正常膀胱尿脱落细胞cD-NA进行两轮消减杂交及两轮抑制性聚合酶链反应(PCR),使得差异表达的DNA片段得以富集。PCR产物与T/A载体连接并转化大肠杆菌JM109构建成差异表达基因的cDNA消减文库。文库扩增后,随机挑取克隆进行酶切、测序及同源性分析。结果:PCR鉴定有317个克隆载有主要在200～900bp之间呈随机分布的插入片段,片段插入率达82.6%,证实建库成功。对20个质粒测序结果经同源性比对分析,其中20个片段源于17个已知基因,1个克隆在GenBank中未检索到与其有相似性的基因序列,表明它们可能为BTCC差异表达的新基因。结论:该消减杂交文库质量可靠,其成功构建为进一步筛选、克隆BTCC差异表达基因提供了依据。     

梗阻性黄疸手术应用单纯全麻与全麻复合硬膜外阻滞疗效比较

目的比较单纯全麻和全麻复合硬膜外阻滞在梗阻性黄疸手术中对患者血液动力学变化及术后苏醒的影响。方法选择40例梗阻性黄疸患者,随机分成单纯全麻组(全麻组)、全麻复合硬膜外阻滞组(复合组),每组20例。两组全麻方法相同,复合组在全麻基础上是复合低浓度小剂量硬膜外阻滞。记录入室后、插管时、术中、拔管时的HR、MAP,两组病人麻醉维持用药量及清醒拔管时间,术后躁动情况。结果复合组在插管拔管过程中HR、MAP均较全麻组低且平稳(P<0.05),全麻组拔管反应剧烈,在插管、拔管时各项指标与诱导前比较均显著升高(P<0.05)。全麻组全麻用药芬太尼、异氟醚、维库溴铵等用药量均显著多于复合组(P<0.05)。复合组较全麻组清醒快,拔管时间明显早于全麻组(P<0.05)。复合组发生躁动1例,显著低于全麻组的11例,P<0.05)。结论全麻复合硬膜外阻滞更加适合于梗阻性黄疸病人的麻醉。     

低蛋白饮食对环孢素A肾病大鼠肾间质纤维化的作用

Objective To investigate whether low-protein diet has protective effect on the progression of renal interstitial fibrosis in rats with cyclosporine A (CsA)-induced nephropathy. Methods Eighteen male Sprague-Dawley rats were randomly divided into three groups (6 rats in each group). The rats in control group (C group) received common diet; in model group (M group) low-salt diet; in intervention group (Ⅰ group) low-salt and low-protein diet. After diet adaptation period of one week, the rats in C group received subcutaneous injection of olive oil 1 mg/kg daily for 5 weeks, while M group and Ⅰ group subcutaneous injection of CsA (diluted into 25 g/L with olive oil) 1 ml/kg for 5 weeks. All the rats were sacrificed at the end of the 5th week. The food-intake and body weight were measured daily. The creatinine clearance (Ccr) was examined before rats were sacrificed. The semi-quantitative pathological analysis on kidney sections was performed. The mRNA and protein expression of transforming growth factor-β1 (TGF-βI) and type Ⅰ collagen (Col Ⅰ) in kidney tissue was determined with real time PCR and immunohistochemical staining, respectively. Results The food-intake and body weight of rats in M and I groups were significantly lower than those in C group (P<0.05). Compared with C group, the Ccr levels in M and Ⅰ groups were significantly reduced [(0.65±0.15) ml/min, (0.40+0.13) ml/min vs (1.55±0.29) ml/min, P<0.05], the relative fibrosis areas of kidney interstitium in M and I groups were significantly increased (3.60%±0.46%, 3.26%±0.75% vs 0.44%±0.24%, P<0.05), the mRNA and protein expression of TGF-β1 in M and I group was significantly up-regulated (by 2.6 and 3.1 times in mRNA and by 1.5 and 1.6 times in protein, respectively, P<0.05), and the mRNA and protein expression of Col Ⅰ in M and I groups was also significantly up-regulated (by 3.0 and 3.5 times in mRNA and by 2.3 and 2.1 times in protein, respectively, P<0.05). There were no significant differences between M and I groups in every parameters above-mentioned except the rat body weight and Ccr. Both the body weight and Ccr in Ⅰ group were significantly lower than those in M group (P<0.05). Compared with C group, the urine osmotic pressure in M group and in I group were deceased (for M group, P>0.05; for I group, P<0.05). Compared with C group, the serum cholesterol levels in M and I groups were significantly increased (P<0.05), and the serum phosphorus level in I group was significantly decreased (P<0.05). The levels of serum albumin and serum calcium of all three groups had no statistical differences (P>0.05). Conclusion Low-protein diet has no renoprutective effects on the rat model of cyclosporin A nephropathy, on the contrary, may induce body weight loss.     

肾功能损伤分级系统在成人心脏术后的应用

Objective The aim of this study was to evaluate of adilty of two acute renal failure-specific scoring systenms (the classification by Bellome et al and the AKIN criteria) for predicting hospital mortality after cardiac surgery in adult patients. Methods Between October 1 st 2006 to Decemjber 31 st 2006, 509 adult patients who ungerwent coronary artery bypass grafting (CABG) and/ or valve operation were enrolled in this study. The medical data collection included gender, age, types of operation, perioperative he- modynamic parameters, urine output, biochemical parameters and outcome. Renal function was assessed daily according to the classi- ficatinn by Bellomo and the AKIN criteria, respectively. As references, Acure Physiology and Chronic Health Evaluation(APACHE) Ⅱ and Sepsis-related Organ Failure Assessment (SOFA) score were also calculated. Resuits Three hundred and forty-one patients were male (67.0%), and 168 were female (33.0%), mean age was (56.2±12.0) years old. Tnree hundred and nine patieats un- derwent CABG, 182 underwent valve operation and 18 underwent CABG plus valve operation, Mean duration of ventilation support was (20.4±17.7) houra, and the ICU stay was (1.4±1.0) days. Postoperative hospital stay was (13.8±9.1) days. According to the classification by Bellomo., the highest in-hospital mortality was 52.9% in ARFS group. Mahiplicatinn of in-hospital morality rate was abserved (X2 for trend, P<0.01) in 0.4% (non-ARF), 1.2% (stage 1), 12.0% (stal～ 2) and 32.4% (stage 3) of pa- tients based on the AKIN criteria. By applying the area under the receiver operating characteristic ourve, the classification by Bellomo and the AKIN criteria had good discriminative power. Furthering, multivariate logistic regression analysis verified that the Odds Ratio of the AKIN criteria was 5.478 (P =0.028, 95% Confidence Interval 1.027- 24.856), after adjusting for gender and age. Con- clusion Analytical data confinned good discriminative power of both the AKIN criteria and the classification by Bellomo for predicting hospital mortality of adult postoperative patient with ARF.     

超声稀释法监测血液透析患者的内瘘功能

目的对维持性血液透析患者进行内瘘血流量（Qb）及再循环率（R％）测定，了解血液透析患者的内瘘功能状态。方法在患者血液透析开始后30min及结束前1h采用超声稀释法测定内瘘功能。对内瘘血流量过低（Qb〈600ml／min）及过高（Qb〉1800ml／min）的患者进行彩色多普勒超声检查。测量内瘘头静脉内径、桡动脉内径、肱动脉内径及平均血流速度并排除血管腔有无狭窄。结果180例患者中的163例成功进行了内瘘功能测定，3例再循环率〉0，占0．02％。平均Qb（1010．3±662．6）ml／min，40例Qb〈600ml／min（24．5％，低流量组），104例Qb为600-1800ml／min（63.8％，正常流量组），19例Qb〉1800ml／min（11．7％，高流量组）。多普勒超声检查低流量组10例，高流量组7例。两组比较在头静脉内径、桡动脉内径、流速、肱动脉流速方面差异均有统计学意义。结论超声稀释法评定血管内瘘功能有较好的敏感性和特异性，是血管通路（包括自身血管和人造血管）功能监测的一种良好的手段。     

肾脏嗜酸细胞腺瘤6例报告

目的：探讨肾脏嗜酸细胞腺瘤的临床特征。方法：报告6例本病患者的临床资料，5例无临床表现，经B超体检偶然发现“肾脏占位”；1例腰痛、腰部不适、触及腹部包块。CT、IVU等影像学检查均有阳性发现。行肾脏部分切除术2例，根治性肾脏切除术4例。术中快速冷冻病理检查确诊。结果：6例术后均痊愈。随访13～96个月，平均78．6个月，2例随访时间未满5年，4例已超过5年（60个月），均未见肿瘤复发和转移。结论：肾脏嗜酸细胞腺瘤是一种少见的良性肿瘤，多无临床表现；影像学检查均有阳性发现；确诊有赖于病理检查，术中快速冷冻病理检查对确定术式有帮助。     

低蛋白饮食对环孢素A肾病大鼠肾间质纤维化的作用

Objective To investigate whether low-protein diet has protective effect on the progression of renal interstitial fibrosis in rats with cyclosporine A (CsA)-induced nephropathy. Methods Eighteen male Sprague-Dawley rats were randomly divided into three groups (6 rats in each group). The rats in control group (C group) received common diet; in model group (M group) low-salt diet; in intervention group (Ⅰ group) low-salt and low-protein diet. After diet adaptation period of one week, the rats in C group received subcutaneous injection of olive oil 1 mg/kg daily for 5 weeks, while M group and Ⅰ group subcutaneous injection of CsA (diluted into 25 g/L with olive oil) 1 ml/kg for 5 weeks. All the rats were sacrificed at the end of the 5th week. The food-intake and body weight were measured daily. The creatinine clearance (Ccr) was examined before rats were sacrificed. The semi-quantitative pathological analysis on kidney sections was performed. The mRNA and protein expression of transforming growth factor-β1 (TGF-βI) and type Ⅰ collagen (Col Ⅰ) in kidney tissue was determined with real time PCR and immunohistochemical staining, respectively. Results The food-intake and body weight of rats in M and I groups were significantly lower than those in C group (P<0.05). Compared with C group, the Ccr levels in M and Ⅰ groups were significantly reduced [(0.65±0.15) ml/min, (0.40+0.13) ml/min vs (1.55±0.29) ml/min, P<0.05], the relative fibrosis areas of kidney interstitium in M and I groups were significantly increased (3.60%±0.46%, 3.26%±0.75% vs 0.44%±0.24%, P<0.05), the mRNA and protein expression of TGF-β1 in M and I group was significantly up-regulated (by 2.6 and 3.1 times in mRNA and by 1.5 and 1.6 times in protein, respectively, P<0.05), and the mRNA and protein expression of Col Ⅰ in M and I groups was also significantly up-regulated (by 3.0 and 3.5 times in mRNA and by 2.3 and 2.1 times in protein, respectively, P<0.05). There were no significant differences between M and I groups in every parameters above-mentioned except the rat body weight and Ccr. Both the body weight and Ccr in Ⅰ group were significantly lower than those in M group (P<0.05). Compared with C group, the urine osmotic pressure in M group and in I group were deceased (for M group, P>0.05; for I group, P<0.05). Compared with C group, the serum cholesterol levels in M and I groups were significantly increased (P<0.05), and the serum phosphorus level in I group was significantly decreased (P<0.05). The levels of serum albumin and serum calcium of all three groups had no statistical differences (P>0.05). Conclusion Low-protein diet has no renoprutective effects on the rat model of cyclosporin A nephropathy, on the contrary, may induce body weight loss.     

肾细胞癌保留肾单位手术的长期随访

目的 探讨保留肾单位手术治疗肾细胞癌的长期疗效及安全性. 方法 回顾性分析243例保留肾单位手术的肾癌患者临床资料.男159例,女84例,平均年龄58(24～77)岁.肿瘤直径平均3.4(1.1～6.7)cm.孤立肾肾癌3例,双侧肾癌11例.TNM分期T_(1a) 237例,T_(1b) 6例;无区域淋巴结及远处转移,无肾静脉、腔静脉癌栓.术后随访行超声、CT、肾功能等检查.应用KaplanMeier方法行肿瘤特异性生存率估计并进行对数秩检验. 结果 243例中232例获随访,平均随访31(1～147)个月.术后应用干扰素继续治疗52例.死亡4例(因癌死亡1例,非癌死亡3例),总体生存率和肿瘤特异性生存率分别为98.3%和100.0%;肿瘤复发5例,远处转移1例.总并发症发生率为5.6%(13/232). 结论 保留肾单位手术治疗肾细胞癌局部复发率低,远期生存率高,并发症发生率低;可以最大限度地保留功能性肾单位,降低并发慢性肾衰竭的危险性,提高患者生活质量,增加患者满意度.     

胃癌患者腹腔游离癌细胞缝隙连接蛋白43表达的研究

目的 探讨缝隙连接蛋白43（Cx43）在胃癌腹腔转移中的作用.方法 收集75例胃癌患者的胃癌组织、癌旁组织及腹腔冲洗液中游离胃癌细胞,应用免疫组织化学染色和免疫荧光染色检测其中Cx43的表达情况,并分析腹腔脱落胃癌细胞Cx43表达与临床病理特征的关系.结果 Cx43在胃癌及癌旁组织中的阳性表达率分别为33.3%（25/75）和100%（75/75）,差异有统计学意义（P＜0.01）.腹腔脱落胃癌细胞阳性检出率为38.6%（29/75）,脱落胃癌细胞Cx43阳性表达率为72.4%（21/29）,显著高于原发胃癌组织（P＜0.01）.腹腔脱落胃癌细胞Cx43表达与肿瘤浸润深度和组织分化程度有关（均P＜0.05）.结论 Cx43基因可能参与了胃癌腹膜转移的发生机制.     

关节镜在儿童髋关节疾患中的应用

目的探讨关节镜在诊断和治疗儿童髋关节疾患中的应用价值。方法对24例(36髋)儿童髋关节疾患进行关节镜手术。先天性髋关节脱位12例(20髋)在关节镜下行真臼清理、关节囊修整,直视下引导复位术;股骨头骨骺缺血性坏死8例(12髋)在关节镜监视下行钻孔减压和滑膜刨削清理术;化脓性髋关节炎4例(4髋)在关节镜下行清创、灌洗、引流术。结果所有病例均获随访,随访时间8~44个月。12例先天性髋关节脱位均获成功治疗,术后再脱位1例,经手法整复复位。无1例出现股骨头骨骺缺血性坏死、髋关节运动受限或关节僵硬等并发症。股骨头骨骺缺血性坏死和化脓性髋关节炎12例术后疼痛消失,关节活动范围恢复正常。结论关节镜对儿童髋关节疾患具有重要的诊断和治疗价值。