糖尿病妇科手术92例的围手术期处理

2002-01／2004-01Ⅱ型糖尿病(DM)妇科手术患92例，年龄31～73(平均53．4)岁．有明确DM病史53例．病程1／12～21(平均6．9)a，入院后新诊断的DM39例空腹血糖为5．9—18．0(平均9．6)mmol/L，餐后血糖为9．3—22．6(平均16．5)mmol/L，术前准备时间平均为6．4d，术后住院时间平均为11．4d，术前有并存症72例．随机抽取同期住院非DM妇科手术92例，年龄22～65(平均41．5)岁，     

Expression and characterization of Gag protein of HIV-1（CN） in Pichia pastoris

To express the core protein of HIV-1 of Chinese prevalent strain （HIV-1 （CN）） in Pichia pastoris, the fulllength gag gene was inserted into the secretory expression vector pHILS1. Linearized recombinant plasmid pHILGAG by Sail was electrotransformed into the yeast strain GS115, and the yeast transformants were identified by PCR. To induce the interest protein to be expressed, the PCR positive transformants were inoculated in the medium of BMGY and BMMY, mRNA of the strain was detected by RT-PCR, and the expressed protein was analyzed by SDS-PAGE, Western blotting and thin layer scanning. mRNA （1.3 kb） was amplified by RT-PCR. SDS-PAGE and Western blotting analysis showed that the molecular mass of the expressed protein was 55 kD, which was similar to the expected value, and the expressed protein could react with McAb to HIV-1 p24. Thin layer scanning analysis demonstrated that the whole amount of the expressed protein was approximately 13 % of the soluble protein in the supernatant. The recombinant yeast had good genetic stability. The optimal expression conditions of the engineering yeast were as follows： BMMY medium, 80-90% of dissolved oxygen, 1% methanol, and 3-day-cultivation course. Gag proteins were expressed under the optimal expression condition and purified via gel filtration chromatography. The purity of the interest protein was up to 85 %. After the purified proteins were inoculated into BALB/c mice, the anti-HIV-1 antibodies in the immunized mice could be detected by Western blotting.     

The effect of two different bioreactors on the neocartilage formation in type II collagen modified polyester scaffolds seeded with chondrocytes

The effect of dynamic culture conditions on neocartilage formation in type II collagen modified polyester scaffolds was studied. Porcine or human articular chondrocytes were seeded in the scaffolds. The cell-scaffold constructs were cultivated statically, in a rotating-type bioreactor or in a shaker for up to 4 weeks. The cell proliferation, morphology, NO production, synthesis of proteoglycans and collagen, and mechanical properties were evaluated. The results demonstrated that the rotating-type bioreactor promoted the growth of primary porcine chondrocytes, helped to maintain their phenotype, and increased the production of extracellular matrix. The constructs also had the largest dynamic compressive modulus. In the static condition, chondrocytes occupied only the outer margin of the cell-polymer constructs. The poor mass transfer in static condition may have caused a lower pH value in the middle of the constructs and lead further to faster scaffold degradation as well as the weakest neocartilage. Constructs in the shaker produced the highest amount of NO as well as the lowest amount of cells and matrix production. Human or porcine chondrocytes of the second passage seeded in scaffolds were much less viable, with the largest amount of cells and matrix when cultured in rotating-type bioreactors. A larger seeding density was required to form neocartilage from passaged adult chondrocytes.     

N-乙酰半胱氨酸对脑死亡状态巴马小型猪肾脏结构、功能与核转录因子κB表达的影响

目的：观察核转录因子κB活性抑制剂N-乙酰半胱氨酸对脑死亡状态下巴马小型猪肾脏结构、功能与核转录因子κB mRNA其蛋白表达的影响，以期提高脑死亡供肾的肾移植效果。方法：实验于2003—08／2004—12在河南省实验动物中心及河南省病理学重点实验室完成。①实验分组及方法：将15只巴马小型猪按随机数字表法分为3组（n=5），即脑死亡组、N-乙酰半胱氨酸组及对照组。脑死亡组和N-乙酰半胱氨酸组均应用改进的缓慢间断颅内加压法建立脑死亡模型，脑死亡组不行药物干预；N-乙酰半胱氨酸组分别于初次确认脑死亡后1h，12h给予N-乙酰半胱氨酸。对照组动物麻醉后仅行开颅与开关腹手术。②实验评估：分别于首次判定脑死亡后3，6，12，18和24h检测动物血清中尿素氮、肌酐、白细胞介素1β、白细胞介素6、肿瘤坏死因子α水平。于脑死亡后3，6，12及24h开腹取相同部位肾组织，苏木精-伊红染色后观察肾组织结构变化，应用免疫组化染色观察核转录因子κB蛋白的表达水平，应用反转录-聚合酶链反应法检测核转录因子κB mRNA动态变化。结果：15只猪均进入结果分析。①自首次判定脑死亡后12h开始，脑死亡组和N-乙酰半胱氨酸组尿素氮和肌酐水平逐渐升高（P〈0．05），相同时间点比较N-乙酰半胱氨酸组显著低于脑死亡组（P〈0．05）。②自首次判定脑死亡3h开始，脑死亡组及N-乙酰半胱氨酸组白细胞介素1β、白细胞介素6、肿瘤坏死因子α逐渐升高（P〈0．05），相同时间点比较N-乙酰半胱氨酸组显著低于脑死亡组（P〈0．05）。③自脑死亡后3h开始，脑死亡组及N-乙酰半胱氨酸组肾组织NF-κB mRNA其蛋白表达水平逐渐升高（P〈0．05），相同时间点比较N-乙酰半胱氨酸组显著低于脑死亡组（P〈0．05）。④N-乙酰半胱氨酸组和脑死亡组动物脑死亡后12h可见肾脏结构变化，N-乙酰半胱氨酸组变化程度明显轻于脑死亡组。结论：N-乙酰半胱氨酸可能通过抑制核转录因子κB mRNA其蛋白的表达，减少炎症介质的释放，从而保护脑死亡状态下肾脏的功能及结构，提高脑死亡供肾肾移植效果。     

亲体肾及造血干细胞联合移植治疗多发性骨髓瘤并发慢性肾功能衰竭

目的:肾功能不全是多发性骨髓瘤并发肾脏疾病的主要表现且决定其预后,探讨1例多发性骨髓瘤并发慢性肾功能衰竭患者行肾及造血干细胞联合移植的治疗体会。方法:南京中医药大学附属医院于2005-04采用肾及造血干细胞联合移植治疗多发性骨髓瘤并发慢性肾功能衰竭患者1例,男性,46岁,经严格的配型及常规检查,发现其胞兄为适合的亲体供者,经供、受者双方同意,结合病情资料并复习文献,先制订出详细治疗方案,然后行亲体肾及造血干细胞联合移植。结果:①供者术后恢复顺利,术后第7天拆线出院,肾功能正常。②受者肾移植术后第3天,血肌酐降至98.2μmol/L,肌酐清除率为45.7mL/min,肾功能恢复正常;干细胞移植术后第13天,查血红蛋白为54g/L、白细胞为4.0×109L-1、中性粒细胞为1.92×109L-1、血小板为30×109L-1,受者造血功能逐步重建,同时抽血行短片段串联重复的嵌合体监测证实受者造血干细胞基因型为完全供者型,说明受者造血功能和免疫功能重建。结论:肾及造血干细胞联合移植治疗多发性骨髓瘤并发慢性肾功能衰竭是一种可行、有效的方法;供、受者的良好配型是移植进行的必备基本条件;另外,术后初期患者处于完全性免疫缺陷状态,故手术后的消毒、隔离措施和抗感染治疗是移植成功的关键。     

商陆多糖Ⅰ联合白细胞介素2对小鼠脾细胞杀瘤活性的影响

商陆多糖Ⅰ(PAP-I),0.3～3μg·ml^-1和小鼠脾细胞培养3～5d可显著增强其杀伤P815肿瘤细胞活性及IL-2(250～500IU·ml^-1)诱导的LAK细胞活性,最适浓度为1μg·ml^-1。PAP-I及IL-2和脾细胞培养的上清液对P815肿瘤细胞无细胞毒作用,但能增强脾细胞及LAK细胞杀瘤活性。PAP-I,5,10及50mg·kg-1,ip可增强脾细胞杀伤P₈₁₅和L₉₂₉细胞的活性及IL-2诱导的LAK细胞活性。     

小叶买麻藤中买麻藤甲素的结构研究

从小叶买麻藤[Gnetum parvifolium(Warb.)C.Y.Cheng]茎的乙酸乙酯部分得到五个成分,经理化常数测定、光谱(UV,IR,MS,~1HNMR,~(13)CNMR和2D-NMR)分析和衍生物制备,确定买麻藤甲素为一个新的2-苯基苯并呋喃类化合物,结构为2-(3′,5′-二羟基-4′-甲氧基苯基)_-3-甲氧基5-羟基苯并呋喃。另外三个鉴定为异丹叶大黄素、白黎芦醇和β-谷甾醇.还有一个化合物正在鉴定中。     

(-),(+)黄皮酰胺对鼠脑内 NMDA- 受体的影响

用[³H]MK-801放射配体竟争结合法测定了(-),(+)黄皮酰胺对大鼠前脑,海马,皮层等部位突触膜的NMDA-R的作用,以探讨其促智机制。同时用饱和实验分析po给药10d后,小鼠脑内该受体密度的变化。结果表明:(-),(+)黄皮酰胺对脑内各部位的NMDA受体均无特异亲和力。但(-)黄皮酰胺在体给药10d后能使小鼠脑内NMDA受体密度显著增高,并呈一定的量效关系。提示黄皮酰胺的药理作用有光学选择性;(-)黄皮酰胺增加脑内NMDA受体密度为其促智作用提供了重要理论依据。     

全固态乌头碱电化学检测器的研制及其在流动注射分析中的应用

报道一种新型结构的全固态乌头碱电化学检测器的研制及其在流动注射分析中的应用。采用流动注射分析法对川乌、草乌及其中成药(小活络丸)中剧毒成分(双酯型生物碱)进行了测试,方法简便快速,结果同光度法接近。本文还提出了电化学法研究乌头碱水解动力学原理。在pH 6.5,温度98℃的条件下测得乌头碱水解成苯甲酰乌头碱的速度常数为1.36×10^-2min^-1。