创伤性急性肺损伤时兔肺微血管内皮细胞Cx40调控内皮细胞通透性的研究

目的 观察创伤性急性肺损伤(ALI)兔血清对肺微血管内皮细胞连接蛋白40(Cx40)表达和间隙连接通道(GJC)功能的影响,及后者与内皮细胞单层通透性的关系.方法 组织块贴壁法分离、培养兔肺微血管内皮细胞.细胞分为3组,对照组、创伤血清处理组和阻滞剂组.在阻滞剂组,细胞GJC阻滞剂1-庚醇(0.5 mmol/L)加入到培养液中.1 h后,阻滞剂组和创伤血清组的20%胎牛血清的培养液均被更换为含有20%创伤血清的培养液.对照组则只更换培养液.3组细胞继续孵育6 h后,进入实验观察.分别行划痕实验、Cx40蛋白免疫印迹、单层细胞通透性和细胞内Ca2+钙浓度检测.结果 Cx40蛋白印迹见创伤血清组与阻滞剂组蛋白表达水平较对照组有明显降低,且2组之间亦不同,阻滞剂组蛋白水平较创伤血清组低,吸光度和值减少(对照组726293.2±83 684.1,创伤血清组397 798.1±87 145.7,阻滞剂组316977.6±76 325.9,n=4,P＜0.05).3组细胞划痕边缘的细胞均被染料荧光黄(LY)染色,LY在对照组细胞扩散得最远,显色的周边细胞的数量最多,创伤血清组次之,阻滞剂组显色的细胞数量最少,相邻细胞与边缘细胞比值的差异有统计学意义(23±5、11±3比7±2,n=4,P＜0.05).3组细胞伊文思蓝清除率均随着时间的延长而增加,且3组细胞间亦有不同,阻滞剂组清除率高于创伤血清组和对照组.组间差异和时间点差异均有统计学意义(P＜0.05).通过共聚焦显微镜的观察,3组细胞荧光强度不相同,阻滞剂组荧光最强,对照组最弱.创伤血清组和阻滞剂组细胞内Ca2+浓度均显著高于对照组[(494.80±41.94)、(569.80±6.70)nmol/L比(158.80±13.09)nmol/L,P＜0.05].结论 创伤性ALI动物血清抑制肺微血管内皮细胞Cx40表达,进而引起GJC功能下降,导致肺血管内皮细胞单层通透性增加,该机制可能参与了胸部创伤后肺血管通透性增加的形成,促进了伤后急性肺损伤的发病.

Abstract：

Objective To investigate the influence of traumatic acute lung injury (ALI) rabbit serum on gap junction channel ( GJC ) function and connexin40 ( Cx40) expression, and the correlation of GJC and Cx40 with rabbit pulmonary microvascular endothelial cells permeability. Methods Cultured pulmonary microvessel endothelial cells were divided into three groups, control ( Gcontrol) , injured serum (Gserum) , and blocker agent ( Gblocker). GJC function was assessed by scrape-loading and dye transfer techniques. Pulmonary microvessel endothelial cells permeability was measured by Evans blue-labeled albumin transfer, and the expression of Cx40 was measured by Western blotting. Intracellular free calcium concentration was measured by fluo-3am. Results Injured serum decreased GJC function and the level of Cx40,which was aggravated by the GJC blocker (control 726 293. 2 ± 83 684. 1, serum 397 798. 1 ± 87 145. 7, blocker 316 977. 6 ±76 325. 9,n=4,P ＜0. 05). Similarly, pulmonary microvascular endothelial cells permeability was increased significantly in Gserum and Gblocker as compared with Gcontrol (P ＜0. 05). After treatment with injured serum in combination with gap junction blocker in vitro, intracellular free calcium concentration was increased [control (494. 80 ±41.94) nmol/L, serum (569. 80 ±6. 70) nmol/L, blocker (158. 80 ±13. 09) nmol/L, P＜ 0.05]. Conclusion Traumatic ALI rabbit serum inhibits the Cx40 expression and down-regulates GJC function, which contributes to the increase of pulmonary microvascular endothelial cells permeability after ALI This mechanism may participate in the increase of pulmonary vascular permeability and promote the morbidity of ALI after trauma.     

单孔腹腔镜在上尿路手术中的应用与体会（附45例报告）

目的总结单孔腹腔镜在上尿路手术中的应用与体会。方法采用单孔五通道,经后腹腔入路肾上腺腺瘤切除术20例、肾囊肿去顶减压术10例、肾癌根治术10例、输尿管切开取石术5例。记录围手术期并发症、手术时间、出血情况、引流管拔除时间、住院时间等。结果患者手术均取得成功,术中、术后均无明显并发症的发生。结论单孔腹腔镜在泌尿外科上尿路手术中的应用是安全和可行的,随着更多的临床实践、器械改进,单孔腹腔镜手术将有更完善和广阔的应用前景。     

锂剂促进周围神经损伤修复的初步研究

目的 探讨锂剂对周围神经损伤后神经再生的影响.方法 取48只雌性SD大鼠,制作大鼠右侧坐骨神经损伤动物模型,通过腹腔注射氯化锂,在不同时间点观察动物下肢活动情况,检测小腿三头肌神经电生理及肌湿重,并对损伤远端神经纤维的神经丝蛋白(NF200)、单核巨噬细胞抗原(ED1)、P-75和运动终板进行免疫组织化学染色观察.结果 损伤后4周,实验组动物下肢已接近正常行走步态,对照组右侧肢体仍明显跛行;损伤后2周和4周,实验组的复合肌肉动作电位(CMAP)波幅较对照组明显增大,两组间的差异有统计学意义(P＜0.05);损伤后4周、8周,实验组的小腿三头肌重量较对照组明显增大,两组间差异有统计学意义(P＜0.05);损伤后3 d,在距离损伤远端5 mm处,实验组坐骨神经纤维内NF200呈连续丝状染色,而对照组仍然是颗粒状染色;损伤后4周,在神经肌肉接头处,可观察到实验组肌肉运动终板有新生神经纤维支配,而对照组运动终板上无神经纤维支配,两组ED1及P75染色未见明显差别.结论 锂剂可有效促进周围神经损伤后的再生,但其机制仍需进一步研究.

Abstract：

Objective To evaluate the effect of lithium on nerve regeneration after peripheral nerve injury.Methods Sciatic nerve crash injury model was created on the right side of 48 female SD rats.Lithium was administered after the injury by intraperitoneal injection.Locomotion of the lower limbs, electromyography and wet muscle weight of the triceps muscles were measured at different time points after the injury.Changes of NF200, ED1, P-75 and motor end plate at the distal part of the injured nerve were detected by immunohistostaining.Results Four weeks after sciatic nerve crash injury, animals that received lithium injection restored near normal gait, whereas the control animals were limping.Two and four weeks after the injury, the lithium injection group had significantly higher CMAP amplitude than the control group.Four and eight weeks after the injury, the wet muscle weight in lithium injection group was significantly heavier than the control group.Three days after crush injury, continuous NF200 positive fibers were found 5 mm distal to the injury site in the lithium injection group, whereas in the control group, only granular NF200 positive staining was observed Four weeks after crush injury, new innervations to the motor end plate were detected in the neuromuscular junction in the lithium injection group, but not in the control group.No differences in ED1 and P75 staining were detected.Conclusion Lithium could significantly promote axon regeneration after peripheral nerve crush injury.Its mechanism is subject to further investigation.     

环氧合酶-2、第10号染色体同源缺失性磷酸酶-张力蛋白基因在肝胆管结石合并胆管癌组织中的表达及意义

目的 探讨环氧合酶-2(COX-2)、第10号染色体同源缺失性磷酸酶-张力蛋白基因(phosphatase and tensin homolog deleted on chromosome ten,PTEN)在肝胆管结石合并胆管癌(hepatobiliary calculus with cholangiocarcinoma,HCWC)中的表达及其意义,研究COX-2、PTEN与HCWC发生、发展的关系,为HCWC的临床防治提供依据.方法 研究分三组:正常胆管组(肝血管瘤或肝外伤切除标本)10例(A组),肝内结石所在胆管组30例(B组),HCWC组37例(C组).应用免疫组织化学SP法,通用二步法检测各组COX-2、PTEN的表达情况,并对检测结果进行分析.结果 COX-2在A、B、C三组中的表达阳性率分别为10.0%、33.3%和70.3%,癌组织中的表达阳性率明显高于对照组胆管组织(P＜0.01).PTEN在A、B和C三组中的表达阳性率分别为90.0%、80.0%和35.1%,癌组织中的表达阳性率明显低于对照组胆管组织(P＜0.01).在COX-2阳性表达的HCWC组织中,PTEN有较低表达.Kendall相关分析表明HCWC组织中COX-2与PTEN表达呈显著负相关(r=-0.323,P＜0.05).结论 COX-2高表达可能与HCWC形成有关.在HCWC中,COX-2和PTEN表达呈负相关.COX-2高表达、PTEN低表达可能提示HCWC更易发生淋巴转移或肝外转移.

Abstract：

Objective To explore the expressions of cyclooxygenase-2 (COX-2), phosphatase and tensin homolog deleted on chromosome ten (PTEN) in hepatobiliary calculus associated with cholangiocarcinoma (HCWC) and their clinical significance. The relationship between the expressions of COX-2, PTEN and the onset and progression of HCWC was investigated to form an experimental base for the prevention and treatment of HCWC. Methods Thirty seven patients with tumor tissues of HCWC (group C), thirty patients with tissues of bile duct surrounding intrahepatic calculus (group B), and ten patients with normal tissues of bile duct from operations of hemangiomas of liver or liver trauma as the control (group A) were sampled and collected. A two-step immunohistochemistry (SP method) was employed to detect and statistically analyze the expressions of COX-2 and PTEN in each of the 3 groups. Results In groups A, B, C, the positive rate of the expression of COX-2 was 10%,33.3%, and 70.3%, respectively. The positive rates of expression of COX-2 in the carcinoma tissues of HCWC was significantly higher compared with the control group (P＜0. 01). In groups A, B, C the positive rates of the expression of PTEN was 90. 0%, 80. 0%, and 35.0%, respectively. The positive rate of expression of PTEN in the carcinoma tissues of HCWC was significantly lower than the control group (P＜0. 01). The expression of COX-2 was followed by a low expression of PTEN in the tissues of HCWC. Kendall's related analysis showed a strong negative correlation between the expression of COX-2 and PTEN in HCWC (r=-0. 323, P＜0. 05). Conclusions A high expression of COX-2 was related to HCWC. There was a negative correlation between the expressions of COX-2 and PTEN in HCWC. A high expression of COX-2 and a low expression of PTEN suggested a high chance of HCWC in extrahepatic or lymphatic metastasis.     

实验性颅内高压诱发家兔自发脑电非线性特征的变化

目的观察实验性高颅压条件下家兔自发脑电非线性特征的变化,探讨在重力变化引起颅内压改变,进而影响脑功能变化的研究中应用脑电复杂性指标的可能性。方法以家免为实验对象,以灌注法改变颅内压,应用关联维和李亚普诺夫指数等参数估计不同颅内压条件下脑电信号的复杂性特征。结果加压前正常颅内压条件下,动物的自发脑电信号均具有正的李亚普诺夫指数和分数关联维,提示这种信号具有混沌性质。实验性高颅压条件下,李亚普诺夫指数在每例动物都明显降低,提示脑电活动的复杂性降低。颅内压恢复正常值,李亚普诺夫指数的变化具有良好的可逆性,提示有限的高颅压引起的脑电活动的复杂度的变化是可逆的。结论自发脑电信号的李亚普诺夫指数作为监测颅内压变化时脑功能变化的指标,具有一定的应用价值。     

生长抑素联合口服万古霉素治疗肝移植术后早期肠梗阻

目的探讨生长抑素联合口服万古霉素治疗肝移植术后早期肠梗阻的疗效。方法将2005年1月至2006年12月间收治的肝移植术后早期发生肠梗阻的58例患者，根据时间和治疗方法分成A组（2005年1月至2005年12月，31例，给予包括禁食、胃肠减压、纠正水电解质和酸碱平衡紊乱，适当胃肠内、外营养以及应用抗生素的常规治疗）和B组（2006年1月至2006年12月，27例，同A组常规治疗措施外，配合采用生长抑素及口服万古霉素）。结果本组肝移植术后早期术后肠梗阻发生率为13％（58／441）。B组患者恢复肠鸣音及排气、排便的时间分别为（7．1±2．0）d和（8．4±2．4）d，A组则分别为（9．1±3．0）d和（10．8±3．4）d；两组差异有统计学意义（P〈0．05）。B组胃肠减压引流量[（298±58）ml／d]明显比A组[（485±106）ml／d]减少；B组发生菌群失调的比例（55％）比A组（77％）明显降低；两组差异亦均有统计学意义（均P〈0．05）。结论在常规治疗基础上，应用生长抑素联合口服万古霉素治疗肝移植术后早期肠梗阻，可明显改善临床症状，减少肠道菌群失调的发生率。     

经皮肾穿刺微造瘘输尿管镜治疗上尿路结石(附156例报告)

目的探讨经皮肾穿刺微造瘘输尿管镜治疗上尿路结石的方法及疗效。方法采用经皮肾穿刺微造瘘输尿管镜下气压弹道碎石处理上尿路结石156冽。结果156例均一期手术取石,单通道取石126例．双通道取石25例,三通道取石5例。其中1次取石120例,2次取石28冽,3次取石8例。136例结石全部取尽。平均手术时间90min,平均出血量50m1,术后平均住院时间8d。所有病例均无严重手术并发症。结论经皮。肾穿刺微造瘘取石术治疗上尿路结石具有创伤小、并发症少、疗效好、恢复快、结石清除率高等优点．值得推广应用．     

肿胀麻醉技术在常规整形美容手术中的应用研究

目的：探讨在隆乳、除皱、扩张器置入等手术中向剥离层次内注入肿胀液的方法及其临床意义。方法：在22例隆乳、15例扩张器置入、10例额颞部除皱术、25例巨痣切除手术中，向手术区注入适量肿胀液后再剥离，并与未注射肿胀液的28例类似的手术进行比较，分析其临床优势和意义。结果：在局部注射肿胀液的病例中，术中剥离时出血少，术后1～3天疼痛减轻，术后局部瘀血、红肿发生率都大为降低。结论：术中局部注射肿胀液能明显减轻术中出血、术后早期疼痛及减少术后局部瘀血红肿的发生率。     

重组蛇毒纤溶因子修饰聚氨酯人工血管的犬颈动脉置换实验

目的通过动物实验评价重组蛇毒纤溶因子（recombinant fibrinolytic enzyme factor II，rF II）修饰聚氨酯（polyurethane，PU）人工血管的植入效果。方法采用浸渍．沥滤法制备口径4mmPU微孔人工血管，扫描电镜观察血管管壁微孔大小和形态，用rF II修饰人工血管内腔。取20只体重（20±1）kg的雄性杂种犬制作颈动脉2cm缺损模型，随机分为3组：rF II修饰PU组（n=8）、无rF II修饰PU组（n=6）和膨体聚四氟乙烯（expanded polytetrafluoroethlyene，ePTFE）组（n=6），植入相应人工血管以修复缺损。记录术后动物一般情况；计算术后30d和60d的血管通畅率；测量术后60d人工血管内径，并进行组织学检查和扫描电镜观察。结果制得的PU微孔人工血管内径（3．74±0．06）mm，壁厚0．4～0．6mm，密度0．25g／cm^3，孔隙率79．8％，径向动态顺应性为8．57％／100mmHg。人工血管管壁内，微孔分布均匀，呈开孔结构。外层孔径为（140±41）Ima，内层孔径为（100±3）μm，外层／内层的厚度比约2：1，内腔表面孔径为（40±16）μm。术后颈部切口愈合良好，动物均存活，无并发症发生。术后30d及60d血管通畅率：rF II修饰PU组分别为100％及66．7％，无rF II PU组为66．7％及33．3％，ePTFE组为66．7％及0，堵塞的人工血管在吻合处发现血栓。rFII修饰PU组、无rF II修饰PU组及ePTFE组植入前血管内径分别为（3．74±0．06）、（3．74±0．06）、（4．00±0．03）mm；术后60d内径分别为（4．51±0．05）、（4.31±0．24）、（4．43±0．12）mm；3组间植入前后比较差异均无统计学意义（P〉0．05）。rF II修饰PU组组织学观察，植入15d有血浆蛋白在内腔表面沉积；30d后有少量细胞黏附；60d后新内膜形成。新内膜厚度随植入时间增加而变厚；植入后60 d rF II修饰PU组人工血管近端、中点及远端的新内膜厚度分别为（560±22）、（78±5）、（323±31）μm（P〈0．05）。扫描电镜观察，rF II修饰PU组新内膜表面由扁长形细胞组成，其长轴顺着血流方向排列，与正常颈动脉内腔表面形貌相似。结论rF II修饰PU血管内腔可提高纤溶活性，减少血栓栓塞的发生，有利于提高植入血管的通畅率。     

腓骨上端带肌蒂游离移植重建右腕关节1例

患者,男，12岁，因右前臂右腕肿痛活动受限3年，于2005年8月20日入院，查体：生命休征正常，右前臂远端右腕部肿胀明显，右桡骨远端膨大，压痛，叩击痛，右前臂右腕活动受限，右手指血运感觉正常。CT示：右桡骨远端见囊状骨质破坏，可见分隔，其内密度为软组织密度影，周边骨质可见局部不连，周边软组织末见肿缺影，邻近骨骨质受侵破坏（图1）。病理会诊回复报告：右桡骨远端符合软骨黏液性纤维瘤。