先兆子痫患者HLA-DQA1、-DQB1、-DPA1基因多态性

目的:探讨人类白细胞抗原HLA-DQ-A1、DQB1、DPA1基因多态性与先兆子痫发病的关系。方法:采用序列特异性引物技术(PCRSSP)对46例先兆子痫患者和105例正常孕妇及其新生儿进行HLA-DQ-DPA1等位基因分型。结果:所有标本共检出11种HLADQA1基因表型、16种HLADQB1基因表型、6种HLADPA1基因表型。先兆子痫患者HLA-DQ-B10301基因频率高于正常孕妇,差异有显著性(Pc=0.032,RR=2.43,AR=0.30),其余各基因表型频率两组比较差异均无显著性。结论:HLADQB10301基因可能是一种先兆子痫发病的易感基因。     

经皮腔内室间隔心肌消融术对心电指标的影响

目的研究肥厚梗阻型心肌病患者经皮腔内室间隔心肌消融术对心电指标的影响。方法对50例肥厚梗阻型心肌病患者行经皮腔内室间隔心肌消融术,记录术前、术中和术后出现的心律失常类型,配对分析术前、术后心电指标的变化。结果术后与术前相比,QRS时限[(122.0±24.0)ms对(97.3±15.5)ms,P=0.000]明显延长,QTc[(469.3±32.2)ms对(434.3±41.5)ms,P=0.004]、PR间期[(169±26)ms对(162±24)ms,P=0.044]稍延长。术中心律失常发生率分别为:右束支传导阻滞70%(35/50),左束支传导阻滞8%(4/50),一过性AVB38%(19/50),频发室性早搏24%(12/50),短阵室性心动过速24%(12/50);未发生持续性室性心动过速和室颤。术后心律失常发生率分别为:右束支传导阻滞56%(28/50),左束支传导阻滞8%(4/50),交界区性心动过速4%(2/50),频发室性早搏16%(8/50),短阵室性心动过速8%(4/50)。无永久性起搏器植入及死亡病例。结论经皮腔内室间隔心肌消融术致心律失常的发生率高,右束支传导阻滞最为常见。严格选择适应证后谨慎地行PTSMA术是安全、可行的。     

肝纤维化门静脉压力改变与血清ET-1/NO关系的实验研究

目的制作兔肝纤维化模型，观察血管活性物质与门静脉压力变化的关系。方法40只家兔采用口服硫代乙酰胺方法造模，分别于造模第8、12、16、20周时检测内皮素（ET-1）及NO浓度，超声观察胆囊壁及肝脏血流动力学指标，直接穿刺测量不同时期门静脉压力。分析血管活性物质与肝脏血流动力学指标及门静脉压力变化间的关系。结果胆囊壁增厚是肝纤维化阶段的二维超声表现；随着纤维化程度的加重，血清ET-1、NO浓度逐渐增加，且以ET-1／N0比值增加更为明显；门静脉内压力、肠系膜上动脉、脾动脉搏动指数（PI）随纤维化程度逐渐增加，实验组与对照组间差异有统计学意义（P〈0．05），且与ET-1／NO值呈正相关（P〈0．01）。结论通过超声可无创检测肝脏血流动力学变化，对肝纤维化临床诊断及治疗效果的观察，具有积极的作用。     

HIV-1C亚型nef基因构建的DNA质粒诱导小鼠免疫反应的研究

目的构建表达中国流行株HIV-1C亚型调控nef基因的重组质粒pVAX-nef，并免疫BALB／c小鼠，观察其免疫效果，为探索新型HIV DNA疫苗提供数据。方法利用分子生物学技术，将nef基因克隆到pVAX，并在体外进行表达与鉴定。以纯化的Nef蛋白作为包被抗原，用ELISA检测其体液免疫反应，用ELISPOT检测其细胞免疫反应。结果重组质粒pVAX-nef成功构建。接种小鼠后2周，ELISPOT结果显示产生了针对HIV特异的CD4和CD8细胞抗原表位的免疫应答，且与免疫剂量存在一定的正相关性。ELISA实验诱导产生了抗HIV-1特异性抗体，其中40μg免疫组诱导的抗体水平最高。结论重组质粒pVAX-nef免疫小鼠后可有效地诱导机体产生细胞免疫和体液免疫反应。     

钙依赖性膜磷脂结合蛋白Annexin Ⅱ对大鼠脊髓横断损伤的作用

目的:观察钙依赖性膜磷脂结合蛋白AnnexinⅡ在体内实验中对大鼠脊髓损伤的作用。方法:在成年大鼠全横断脊髓损伤局部微量注射AnnexinⅡ,观察其对神经细胞存活、神经纤维生长以及损伤后的功能恢复的影响。结果:在脊髓横断损伤6周后,损伤局部微量注射AnnexinⅡ组斜板实验平均角度和运动评分结果比对照组均有非常明显增高,脊髓损伤处的空洞面积非常显著减小;同时,损伤节段以上荧光金标记神经细胞数目也较对照组非常明显增多。但与神经营养因子的神经保护效应相比尚存在明显差异。结论:AnnexinⅡ蛋白具有减小局部损伤空洞面积,保护损伤的神经元和神经纤维并防止其变性和死亡的效应,可使脊髓损伤大鼠的功能得到一定程度的恢复。     

先天性淋巴管发育畸形的临床特点分析

目的分析先天性淋巴管发育畸形(LM)的临床特点,提高临床医师对本病的认识。方法收集14例LM患者的临床资料,并进行相关文献复习。结果①14例患者中原发性小肠淋巴管扩张症(PIL)6例,合并乳糜腹水1例、乳糜心包积液1例、皮肤及胃肠道血管畸形1例;原发性淋巴性水肿(PL)8例,合并乳糜胸水3例、乳糜腹水1例;②PIL的主要临床表现为双下肢对称性水肿、腹泻、血淋巴细胞减少及非选择性低蛋白血症;PL的主要临床表现为双下肢不对称性水肿及呼吸系统症状;③淋巴管造影或同位素淋巴管显像均显示异常,表现为淋巴回流异常;④PIL患者内镜检查显示,十二指肠、小肠黏膜呈白点状改变;活检病理检查可见小肠黏膜及黏膜下扩张淋巴管;⑤14例患者中6例行外科淋巴管-静脉吻合术后好转,8例予以MCT饮食、对症支持治疗及局部物理加压治疗。结论LM的诊断应结合淋巴管造影、同位素检查及病理学诊断,治疗上应内、外科综合治疗。     

Bioreactor expansion of human adult bone marrow-derived mesenchymal stem cells

Supplementation of mesenchymal stem cells (MSCs) during hematopoietic stem cell (HSC) transplantation alleviates complications such as graft-versus-host disease, leading to a speedy recovery of hematopoiesis. To meet this clinical demand, a fast MSC expansion method is required. In the present study, we examined the feasibility of using a rotary bioreactor system to expand MSCs from isolated bone marrow mononuclear cells. The cells were cultured in a rotary bioreactor with Myelocult medium containing a combination of supplementary factors, including stem cell factor and interleukin-3 and -6. After 8 days of culture, total cell numbers, Stro-1(+)CD44(+)CD34(-) MSCs, and CD34(+)CD44(+)Stro-1(-) HSCs were increased 9-, 29-, and 8-fold, respectively. Colony-forming efficiency-fibroblast per day of the bioreactor-treated cells was 1.44-fold higher than that of the cells without bioreactor treatment. The bioreactor-expanded MSCs showed expression of primitive MSC markers endoglin (SH2) and vimentin, whereas markers associated with lineage differentiation, including osteocalcin (osteogenesis), type II collagen (chondrogenesis), and C/EBP-alpha (CCAAT/enhancer-binding protein-alpha) (adipogenesis), were not detected. Upon induction, the bioreactor-expanded MSCs were able to differentiate into osteoblasts, chondrocytes, and adipocytes. We conclude that the rotary bioreactor with the modified Myelocult medium reported in this study may be used to rapidly expand MSCs.     

Expression patterns of ER-alpha, PR, HER-2/neu, and EGFR in different cell origin subtypes of high grade and non-high grade ductal carcinoma in situ

We have previously reported that high grade and non-high grade ductal carcinoma in situ (DCIS) of the breast can be subdivided into 3 cell origin subtypes (luminal, basal/stem, and null), and that high grade DCIS is more frequently associated with basal/stem cell subtypes compared to non-high grade DCIS. Here we refine the relationships between these 3 subtypes and the expression patterns of estrogen receptor-alpha (ER-alpha), progesterone receptor (PR), HER-2/neu, and epidermal growth factor receptor (ERFR) in 53 cases of non-high grade and 46 cases of high nuclear grade DCIS. Using a panel of antibodies to ER-alpha, PR, HER-2/neu, and EGFR, along with cytokeratin (CK) markers (CK5/6, CK8, CK14, CK17, and CK18), we found that all 3 cell origin subtypes can express ER-alpha and PR, and their expression is higher in non-high grade DCIS than in high grade DCIS; the expression of HER-2/neu is associated with luminal subtype only in non-high grade DCIS, but can be seen in all 3 subtypes in high grade DCIS; the expression of EGFR is low and is present only in luminal cell subtypes in both high and non-high grade DCIS. Basal/ stem cell and null cell subtypes occur in younger patients in non-high grade DCIS compared to high grade DCIS. In conclusion, the expression patterns of ER-alpha, PR, HER-2/neu, and EGFR are markedly different in different cell origin subtypes of both high grade and non-high grade DCIS, suggesting that cell origin subtypes as well as nuclear grade contribute to the biological and molecular heterogeneity of DCIS.     

大鼠癫痫敏感性增强过程中c-jun与GFAP基因表达的关系

本文利用免疫细胞化学技术研究了海人酸(KA)致大鼠癫痫敏感性长期增强过程中,原癌基因c-jun的表达规律及其与星形胶质细胞增生、星形胶质细胞的标志物-神经胶质原纤维酸性蛋白(GFAP)基因表达的关系。一次性给予惊厥剂量(10mg/kg,i.p.)的KA后,海马CA1区出现c-jun的一过性高表达,之后呈持续表达状态直至给予KA后6个月。c-jun的表达明显地早于给予KA 2 d后方开始出现的海马内CA1区锥体细胞层神经元的死亡和C-JUN免疫反应阳性的星形胶质细胞的增生。星形胶质细胞GFAP基因的表达与其细胞内的c-jun表达的时间和部位一致。结果提示,c-jun在癫痫敏感性长期增强过程中持续表达,并可能参与介导神经元死亡和维持星形胶质细胞增生以及调控其细胞内GFAP基因的表达。     

比较两种方法制备脱细胞小血管支架

目的:比较0.5%Triton X-100 0.05%NH4OH和酶法制备脱细胞小血管支架的效果.方法:分别用含0.5%Triton X-100 0.05%NH4OH处理3 d或1.0H Triton X-100 0.125%胰蛋白酶 Dnase Rnase孵育48 h,脱去犬股静脉中的细胞成分,50Co辐照消毒,血清浸泡24 h,将平滑肌细胞和内皮细胞接种到脱细胞支架中;进行H-E染色、胶原纤维和弹力纤维染色,扫描电镜观察及力学检测.结果:0.5%Triton X-100 0.05%NH4OH法完全地脱去了血管细胞;细胞外基质较完整地保留下来,其形态结构与脱细胞前无明显改变;见种植细胞在支架内生长良好,连成片,支架具有良好的生物相容性;力学结果显示其弹性回复率和最大断裂强度好于酶组.结论:两种方法比较,0.5%Triton X-100 0.05%NH4OH法简便易行,成本低,脱细胞效果好,组织相容性佳,对力学性状影响小,是比较理想的制备脱细胞小血管支架的方法.