骨髓单个核细胞移植可影响急性心肌梗死后心室重构

目的 观察同种异体骨髓单个核细胞(bone marrow mononuclear cells, BM-MNCs)移植对大鼠急性心肌梗死后左心室重构的影响.方法 健康雄性Wistar大鼠40只,随机均分为对照组和实验组.分别将制备的培养基和BM-MNCs悬液经心外膜下种植对照组和实验组梗死心肌周围.结果 移植术后4周,实验组左心室非梗死区心肌骨桥蛋白基因(OPN mRNA)、Ⅰ型胶原和AngⅡ含量均显著低于对照组(P均<0.05).Pearson直线相关分析表明,实验组和对照组心肌AngⅡ、Ⅰ型胶原与OPN mRNA三者之间两两均呈正相关(P均<0.05).结论 同种异体骨髓单个核细胞移植可能通过抑制大鼠心肌Ⅰ型胶原合成,下调心肌AngⅡ和OPN基因表达,减轻左心室重构.     

应力刺激结合温经活血汤治疗骨折延迟愈合疗效观察

目的:观察应力刺激结合温经活血汤治疗骨折延迟愈合的临床疗效。方法:40例随机分成两组各20例,两组均行应力刺激。治疗组给予温经活血汤治疗、对照组给予接骨七厘片治疗。结果:治疗4周、8周两组血清ALP、BGP指标比较差异有统计学意义(P<0.05),治疗4周、8周和治疗结束后4周骨痂评分两组比较差异均有统计学意义(P<0.05),总体疗效治疗组优于对照组(P<0.05)。结论:应力刺激结合温经活血汤治疗骨折延迟愈合疗效较好。     

Effects of portal venous arterialization on acute occlusion of hepatic artery in rats

Background A fatal complication after liver transplantation is anastomotic embolization of the hepatic artery. In order to solve this problem, the portal venous arterialization （PVA） is used to reconstruct the hepatic arterial blood flow. The purpose of this study was to investigate the influence of PVA on rats with acute occlusion of hepatic artery.
Methods Rat PVA models were established and then randomly divided into Group 1 （control group）, Group 2 （jaundice group）, Group 3 （bile duct recanalization group）, and Group 4 （portal vein arterilization group）. Recanalization of the common bile duct and PVA were performed 5 days after bile duct ligation in the rats. The influence of the PVA on general conditions, hepatic changes of structure and function, portal vein pressure and hepatic micrangium were observed for one month.
Results Five days after common bile duct ligation the serum bilirubin, transaminase and alkaline phosphatase levels were significantly increased. Compared with group 1, there was a statistically significant difference （P 〈0.01）. These rats then underwent bile duct recanalization and PVA. After a month, the liver functions and microscopic structures completely returned to normal and, compared with group 1, there was no statistically significant difference in portal vein pressure （P 〉0.05）. Vascular casting samples showed that hepatic sinusoids were slightly thicker and more filled than normal ones and although they had some deformations, the hepatic sinusoids were still distributed around the central vein in radial form.
Conclusion Within a month after operation, bile duct recanalization and PVA do not show obvious adverse effects on liver hemodynamics and hepatic micrangium, and the liver function and microscopic structure can return to normal.     

氟烷和七氟醚对缺血心肌功能和代谢及Ca2+-ATP酶活性的影响

目的: 研究氟烷、七氟醚(1.5MAC)对缺血心肌的影响。方法: 应用离体大鼠心脏Langendorff逆行灌注模型研究氟烷、七氟醚对心肌缺血前心率(HR)、左室舒张末期压力(LVEDP)、左室发展压(LVDP)、左室压力升高速率(+dp/dt)、左室压力下降速率(-dp/dt)和冠脉流量(CF)的影响,测定缺血前、缺血10min、缺血25min3个不同时间的心肌ATP含量、Ca²⁺-ATP酶活性,同时记录缺血间期左室内压的变化情况。结果: 七氟醚显著增加正常离体心脏的CF,氟烷、七氟醚均不同程度地抑制心肌收缩功能和Ca²⁺-ATP酶活性,能够增加正常心肌的能量贮备。缺血10min时,二药能够减缓心肌ATP含量及Ca²⁺-ATP酶活性的下降,氟烷的作用比较明显。缺血间期,氟烷明显推迟缺血性挛缩的起始时间,降低挛缩幅度。结论: 氟烷的抗缺血损伤作用优于七氟醚,延缓缺血期心肌ATP含量及Ca²⁺-ATP酶活性的下降可能是氟烷抗缺血损伤作用的重要机制之一。     

Expression, purification and molecular modeling of recombinant fibrinogenase [IV], a metalloproteinase from Deinakistrodon acutus venom.

A novel metalloproteinase, recombinant fibrinogenase IV (rFIV(a)), was expressed and purified from Deinakistrodon acutus venom. It was a single chain protein with an apparent molecular weight 27 kDa and an isoeletric point of pH 7.1. RFIV(a) cleaved preferentially the Aalpha-chain and also cleaved Bbeta, gamma-chains of fibrinogen when the incubation time was prolonged. The proteolytic activity was inhibited by EDTA, l-cysteine, and DTT, indicating rFIV(a) was a metalloproteinase requiring disulfide bonds for its activity. It kept above 85% of the initial activity from pH 4.5-11, showed an equal maximum activity at the temperature range from 30 to 50 degrees C, and was inactivated by Zn2+, Cu2+ and Cd2+. Homology modeling of rFIV(a) showed that two highly conserved disulfide bonds (Cys159-Cys164 and Cys117-Cys197) was maintained from its structure, and it exhibited the characteristic conserved motif H142E143XXH146XXGXXH152, whose three histidine residues were involved in binding of the catalytically essential zinc ion. This work demonstrates the expression, purification and characterization of recombinant fibrinogenase IV, which belongs to class P-I metalloproteinase from D. acutus venom.     

心外科重症监护病房环境卫生学监测分析

目的　了解心外科重症监护病房外环境中常见的病原菌及其分布情况,以便有针对性采取预防措施,减少交叉感染的发生。方法　2 0 0 4年8月～10月对心外科重症监护病房物体表面、空气、工作人员手和鼻前庭、湿化瓶、呼吸机管道进行生物监测。结果　分离出各类细菌5 6种,32 4株,其中革兰氏阳性菌2 2 3株,革兰氏阴性菌10 1株,主要细菌有葡萄球菌、鲍曼不动杆菌、肺炎克雷伯菌、铜绿假单胞菌。大多分布于输液泵、门把拉手、电话、电脑键盘、病历夹表面及工作人员鼻腔和手。结论　严格执行消毒隔离措施是预防和控制环境因素导致重症监护病房病人获得医院感染的有效方法。     

C57BL/6N小鼠早期视网膜变性及小胶质细胞活化状态研究

目的：观察C57BL/6N(Crb1^rd8/rd8)小鼠早期视网膜变性以及小胶质细胞的活化情况。

方法：取雄性SPF级C57BL/6N小鼠15只、C57BL/6J小鼠15只,正常饲养。分别在入组时,入组4、8、12 wk,使用Micron-Ⅲ小动物视网膜影像系统进行双眼彩色眼底照相检查计算病变数量、病变面积。观察结束后处死小鼠,摘取右侧眼球制备视网膜组织切片,进行HE染色后光镜下观察视网膜组织形态; 采用免疫组织化学染色分析两组小鼠视网膜CX3CR1的表达水平及位置。摘取左侧眼球分离视网膜,使用Western-Blot检测CD86、CD206的表达情况,使用电化学发光法测定视网膜中IL-1β、IL-6、TNF-α、IL-4、IL-10炎性因子含量水平。

结果：眼底彩照结果显示在入组4、8、12 wk,C57BL/6N组眼底病变数量均较入组时显著增加,与C57BL/6J组同时间点变化量比较均有差异(均P<0.05); 眼底病变面积变化量两组间入组12 wk有差异(P<0.05),各组内变化量比较均无差异(均P>0.05); 视网膜组织HE染色示：C57BL/6N组视网膜结构异常,细胞排列疏松、紊乱,光感受器层向视网膜内侧明显的玻璃膜疣样凸起,C57BL/6J组视网膜结构清晰,细胞排列有序,无明显异常; 免疫组化结果示：C57BL/6N组视网膜中CX3CR1高表达于神经节细胞层、内外丛状层、感光细胞层及病灶处位置,平均光密度为0.285±0.056,C57BL/6J组为0.189±0.084(P<0.05); Western-Blot结果显示：与C57BL/6J组相比,C57BL/6N组视网膜CD86、CD206蛋白有不同程度升高,两组CD86蛋白表达有差异(P<0.05); 细胞因子检测结果显示：C57BL/6N组IL-1β、TNF-α水平显著高于C57BL/6J组,而IL-10含量则明显较低(均P<0.05)。

结论：C57BL/6N(Crb1^rd8/rd8)小鼠视网膜变性进展缓慢,随年龄呈进行性加重,病灶部位视网膜结构紊乱,伴有以M1型极化为主的小胶质细胞浸润。     

全反式视黄酸通过RARα抑制缺氧缺糖引起的原代神经元凋亡

目的:探讨全反式视黄酸（all-transretinoicacid,ATRA）是否能够抑制缺氧缺糖损伤（oxygenandglucose-deprivation,OGD）诱导的原代神经元凋亡。方法:采用原代神经元作为研究对象,分为对照组、OGD组、OGD+ATRA组;体外分离培养原代神经元,用神经元特异性烯醇化酶（neuron-specificenolase,NSE）鉴定神经元;倒置显微镜观察OGD损伤前后细胞形态变化;流式细胞技术研究OGD损伤后各组原代神经元凋亡水平;Real-timePCR检测视黄酸核受体α（retinoicacidreceptoralpha,RARα）、转录生长因子β1（transforminggrowthfactor-β1,TGF-β1）、转录生长因子β2（transforminggrowthfactor-β2,TGF-β2）、含半胱氨酸的天冬氨酸蛋白水解酶-3（cysteinylaspartatespecificproteinase,Caspase-3）mRNA表达水平;Westernblot印迹检测RARα、TGF-β、Caspase-3蛋白表达水平。结果:成功培养原代神经元;原代神经元OGD损伤后,倒置显微镜观察发现OGD组死亡的原代神经元较OGD+ATRA组明显增多,细胞更加杂乱;流式细胞计数发现OGD+ATRA组（26.11±2.96）%细胞凋亡率较OGD组（52.14±5.23）%明显降低（P=0.025）;OGD+ATRA组RARαmRNA表达水平（1.52±0.43）和蛋白表达水平（0.22±0.02）较OGD组的mRNA表达水平（0.49±013）和蛋白表达水平（0.10±0.02）均明显增高（P=0.006,P=0.005）,而OGD+ATRA组TGF-β1、TGF-β2mRNA表达水平（0.45±0.10,0.41±0.08）分别较OGD组（0.81±0.15,0.98±0.15）明显降低（P=0.034,P=0.018）,而且OGD+ATRA组TGF-β、Caspase-3蛋白表达水平（0.22±0.04,0.18±0.03）分别较OGD组（0.39±0.05,0.47±0.06）也明显降低（P=0.016,P=0.002）,OGD+ATRA组（1.08±0.49）与OGD组（1.00±0.15）Caspase-3mRNA表达水平无明显差异（P=0.148）。结论:ATRA可以激活其受体RARα进而下调TGF-β,抑制凋亡因子Caspase-3的表达,最终抑制OGD损伤引起的原代神经元凋亡。     

复方浙贝浸膏对人结肠癌耐奥沙利铂细胞株HCT-116/L-OHP裸鼠移植瘤及凋亡蛋白表达的影响

目的观察复方浙贝浸膏(CompoundZhebeiExtract,CZBE)对人结肠癌耐奥沙利铂(L-OHP)细胞株(HCT-116/L-OHP)裸鼠移植瘤体内抑制作用,以及对移植瘤细胞相关凋亡蛋白表达的影响。方法将人结肠癌耐L-OHP细胞HCT-116/L-OHP接种于BALB/c裸鼠右腋前皮下构建结肠癌耐L-OHP移植瘤模型,成模后按随机数字表法将其分成肿瘤模型对照组、阳性药L-OHP组(3mg/kg)、CZBE高剂量(CZBE10g/kg)联合L-OHP组、CZBE中剂量(CZBE5g/kg)联合L-OHP组、CZBE低剂量(CZBE2.5g/kg)联合L-OHP组,分组当天给药。给药方法:CZBE,灌胃,每日1次;L-OHP,3mg/kg,腹腔注射,隔日1次。连续用药2周。实验结束后处死小鼠,称体重后完整剥离瘤体并称瘤重,计算肿瘤抑制率。将瘤块组织制成切片,通过免疫组化结合ImageProPlus7.0彩色图像分析软件及SPSS21.0软件,分析CZBE对HCT-116/L-OHP移植瘤细胞凋亡蛋白表达的影响。结果CZBE各剂量联合L-OHP均能有效抑制HCT-116/L-OHP裸鼠移植瘤体积的增长。CZBE高剂量联合L-OHP组肿瘤抑制率为45.031％,与阳性药L-OHP组比较差异有统计学意义(P<0.05),CZBE中剂量联合L-OHP组肿瘤抑制率为37.669％,复方浙贝浸膏低剂量联合L-OHP组肿瘤抑制率为32.147％,阳性对照L-OHP组的肿瘤抑制率为27.239％,并延长移植瘤裸鼠的生存期。与阳性药L-OHP组相比,CZBE低剂量联合L-OHP组移植瘤细胞B淋巴细胞瘤-2基因(Bcl-2)的累积光密度/面积(IOD/Area)值降低(P<0.05),CZBE中、高剂量联合L-OHP组移植瘤细胞Bcl-2的IOD/Area值降低(P<0.01)。CZBE中剂量联合L-OHP组移植瘤细胞Bcl-2相关x蛋白(Bax)的IOD/Area值升高(P<0.05),CZBE高剂量联合L-OHP组移植瘤细胞Bax的IOD/Area值升高(P<0.01)。结论CZBE联合L-OHP能提高人结肠癌耐L-OHP细胞株移植瘤的肿瘤抑制率,并延长人结肠癌耐奥沙利铂移植瘤裸鼠的生存期,其机制可能是通过调节Bax/Bcl-2凋亡途径逆转肿瘤多药耐药性,促进肿瘤细胞凋亡。