凋亡素原核表达载体的构建和表达

目的 构建凋亡素原核表达系统，以制备抗原物质凋亡素融合蛋白。方法 通过PCR方法，以pcDNA-VP3质粒为模板，扩增出凋亡素VP3基因。将其克隆到原核表达载体pET-DsbA的多克隆位点，构建成凋亡素的高效原核表达栽体pET-DsbA-VP3，将该质粒转化到大肠杆菌E．coliBL21(DE3)plysS中，以异丙基硫代-β-D-半乳糖苷(isopropylthio-β-D-galactoside，IPTG)对其进行诱导表达，聚丙烯酰胺凝胶电泳分析目的蛋白。结果 转化有凋亡素原核表达载体pET-DsbA-VP3的大肠杆菌E．coliBL21(DE3)plysS经IPTG诱导后，聚丙烯酰胺凝胶电泳出现38300的目的蛋白条带。结论 凋亡素原核表达栽体pET-DsbA-VP3能高效表达出凋亡素融合蛋白。     

高效液相色谱法测定丹皮酚-β-环糊精包合物片的含量

目的 采用高效液相色谱法（HPLC法）测定丹皮酚-β-CD片中丹皮酚的含量。方法 以十八烷基硅烷键舍硅胶为填充剂，甲醇-水（60：40）为流动相，检测波长为274nm。结果 丹皮酚进样量在0．20～1．0μg范围内与峰面积线性关系良好，r=0、9999，平均加样回收率为100．9％，RSD：0．38％（n=6）。结论 IIPLC法方便快捷，准确，灵敏度高，重现性好。     

经喉内镜手术治疗喉部疾病的并发症分析

目的：分析经喉内镜手术后并发症的发生原冈及其防治措施。方法：使用从德国STORZ公司引进的国内首套喉内镜手术系统，治疗包括声带小结、声带息肉、喉乳头状瘤、声带囊肿、会厌囊肿、喉部炎性假瘤、喉部血管瘤等58例患者，并随访2～36个月，观察主要并发症并作相应处理。结果：58例患者中发音正常者54例，发音功能明显好转者4例。主要并发症有：牙齿松动2例，牙齿脱落1例，声带粘连1例，舌体麻木1例，舌运动障碍1例，术后拔管时喉痉挛1例。结论：经喉内镜手术可住微创的前提下彻底清除病灶，并最大限度地保留发音功能，只要术前严格掌握适应证，术中精细操作，其并发症是可以避免的。     

恶性血液病患者血清红细胞生成素的水平及其临床意义

目的 探讨恶性血液病患者血清红细胞生成素(Epo)的含量及其意义。方法 采用夹心酶联免疫法测定40例恶性血液病患者及18例健康人血清Epo，同时检测其血红蛋白及红细胞数。结果 恶性血液病中，急性白血病及多发性骨髓瘤的Epo较正常人明显增高(P＜0．001)，其血红蛋白值则明显低于正常人(P＜0．001)，慢性白血病及淋巴瘤的Epo也比正常人高(P分别为＜0．002及＜0．05)，但其血红蛋白值则与正常人无显著差异(P＞0．05)。各组病人血清Epo水平均与血红蛋白及红细胞数呈负相关。结论 恶性血液病血清Epo水平较对照组明显升高，并与血红蛋白及红细胞数呈负相关。除贫血、缺氧，导致体内Epo活性反馈，使患者Epo水平增高外，可能还和红系祖细胞利用Epo减少有关。rhEpo对Epo基础值不太高的某些恶性血液病伴发的贫血是一种有效的治疗措施。     

武汉市民遗体捐赠意愿因素的问卷调查与分析

目的:了解武汉市民的捐献意愿及影响不同阶层市民捐献意愿的主要因素.方法:对武汉市武昌区10个社区的居民进行随机访问,并发放问卷,然后对问卷进行对比研究分析总结.内容:法制因素、情感因素、传统观念、社会舆论等.结果:法制因素和情感因素为最显著因素.结论:武汉市对遗体捐献进行立法是非常正确的,但要想收到预期的效果还应加大遗体捐献知识的宣传普及力度.     

精浆蛋白电泳分析的临床意义

目的为了初步了解男性精浆中的蛋白电泳谱以及蛋白谱与男性生育能力的相关关系。方法通过应用SPIFE3000全自动电泳分析仪对20例对照组(生育组)与140例观察组(不育组)的精浆进行蛋白分离(琼脂糖区带电泳法),建立其精浆蛋白质的特征电泳图谱,并进行对比分析。结果精浆蛋白经酸蓝染色后,区带清晰,从阳极到阴极分为A、B、C、D、E 5条带,电泳各区带百分比含量结果经方差分析,发现不育组与生育组A、C、D、E百分比含量变化有统计学意义(P<0.01),且无精子症患者精浆的蛋白谱有特异的"双峰"特点。结论精浆蛋白区带电泳分析可作为不育症的辅助诊断指标,且将是对男性不育诊断手段的有力补充。     

对常规体外受精失败后补行卵胞浆内单精子注射的探讨

目的 探讨对可疑受精障碍的常规体外受精(IVF)周期在授精失败后补行卵胞浆内单精子注射(reacue ICSI)的时机及临床效果.方法 对2006年1月至2007年6月在我中心接受常规IVF助孕治疗,可能有受精障碍的13例患者,取卵后6h授精,4h后观察,发现所有卵母细胞均未排出第二极体,立即行补救ICSI.结果 13个周期共154个卵母细胞,其中M Ⅱ期142个;未行补救ICSI的46个均未受精;补救ICSI的96个,受精89个,受精率92.7%,移植13个周期29个胚胎,临床妊娠1例双胎和5例单胎,种植率24.1%.结论 常规IVF受精后4h行补救ICSI可以获得较好的受精率和妊娠率.     

Application of trans-abdominal chorionic villus sampling in prenatal diagnosis of chromosomal diseases in first trimester of gestation

Objective:To evaluate the feasibility and safety of prenatal diagnosis by traneabdominal chorionic villus sam-pling(TA-CVS)via the guidance of B-mode ultrasound in the first trimester of gestation.To explore the technique of long time culture and chromosome preparation of villi in early pregnancy.To evaluate the feasibility of the above techniques in the application of the prenatal cytogenetic diagnosis.Methods:One hundred and thirty-five singleton pregnancies at risk were referred from January 2001 to Decem-ber 2007.Results:The average maternal age was 35.2 years.TA-CVS was performed in the 10～13th weeks of gestation and the average gestational age was 10.89 weeks.All attempts at sampling were successful.The rate of operation-associated fetal loss was 0.74%.The failure rate of prenatal diagnosis because of inadequate amount of specimen was 0.The average culture time was 5-7 days.The success rate of the cell culture was 98.5%.No maternal con-temination and bacterial contamination happened.Fifteen cases of abnormal karyotype and one case of confined pla-cantel mosaiciem were diagnosed.Conclusion:TA-CVS appears to be safe and feasible and might to be offered in the prenatal diagnosis in the first trimester of gestation.The technique of long time culture and chromosome preparation of villi is stable and reliable.It is feasible to apply these techniques in the clinical practice of prenatal cytogenetic diagnose in the early pregnancy.     

机械振动的成骨效应研究进展

应力是骨形成的“指导性”因素。其机制是近十年来骨科学、生物医学工程及康复医学领域的研究热点。机械振动是力学刺激形式之一。诸多研究表明：振动具有显著的成骨效应,其在骨质疏松的治疗中将会有广阔的应用前景。但是,现有相关研究中,具有成骨效应的振动频率、强度等参数并不一致,尤其是振动的成骨机制不明,相关报道极少,本文对其作一论述,可为更系统、深入地研究及其在临床中的应用提供理论基础。     

Lack of evidence for fecal-oral transmission of Helicobacter pylori infection in Taiwanese.

BACKGROUND AND PURPOSE: Helicobacter pylori infection is primarily acquired in early childhood and its transmission routes are debated. The aims of this study were to determine the seroprevalence of anti-H. pylori immunoglobulin G (IgG) in Taiwanese and to investigate whether a common mode of transmission could be shared between H. pylori and hepatitis A virus (HAV). METHODS: An enzyme-linked immunosorbent assay (ELISA) was used to investigate the prevalence of H. pylori among 924 healthy volunteers aged less than 40 years, and radioimmunoassay for HAV infection was conducted in 500 subjects from the same population. The kappa statistic was used to measure the difference in positivity for the 2 infections in a subgroup of 500 subjects who had their sera simultaneously tested for anti-H. pylori and anti-HAV antibodies. RESULTS: The seroprevalence of anti-H. pylori IgG was 16.7% in subjects aged < 16 years and 38.3% at ages >or= 16 years. Most children (99%) under the age of 16 were seronegative for HAV. The prevalence of H. pylori infection increased rapidly with age, at 1%/year and 0.8%/year for children and adults, respectively. The rate of H. pylori infection was higher in male (21%) than in female (12%) children (p < 0.01; OR, 2.0; 95% confidence interval, 1.2 to 3.0). No significant difference in seroprevalence between genders was noted in subjects aged >or= 16 years. The agreement in the trend of seropositivity between both infections in the age groups 1 to 10 years and 10 to 20 years was worse than chance (kappa = -0.56) and little better than chance (kappa = 0.01), respectively. CONCLUSIONS: Acquisition of H. pylori infection occurs at a young age, and male children are more likely to develop the infection. The main transmission route of HAV, the fecal-oral route, did not seem to be responsible for H. pylori transmission in this Taiwanese cohort.