Detection of cell apoptosis by MTT assay.]

OBJECTIVE: To study the feasibility of using MTT assay to detect cell apoptosis. METHODS: K562 cells were grown in RPMI 1640 medium supplemented with 10% calf serum and 4 micromol/L arsenic trioxide. Apoptosis was induced in the cultured cells by As2O3, and the cells were detected with optical microscope, DNA gel electrophoresis and MTT staining respectively. RESULT: MTT staining could also accurately detect cell apoptosis, by which the apoptotic cells were easily distinguished from normal cells and dead cells. CONCLUSION: MTT staining is simple, convenient and practical for detecting cell apoptosis.     

人胎盘基因表达谱芯片的初步研究

目的：制备人胎盘基因表达谱芯片，并用于基因差异表达分析。方法：用基因芯片点样仪将胎盘基因文库探讨打印在氨基包被的玻片上制备表达谱芯片。分别提取对照组与三氧化二砷处理的K562细胞总RNA，纯化mRNA后反转录成cDNA，再以限制性显示技术进行荧光标记，将两组荧光标记的样品混合后与芯片进行杂交，杂交后清洗芯片，干燥后对芯片进行扫描，分析杂交结果。结果：建立了较可靠的制备与检测基因表达谱芯片的方法，并筛选出45个差异表达基因片段，结论：制备的人胎盘基因表达谱芯片可有效地应用于基因的差异表达分析。     

ELISA法检测可溶性人类白细胞抗原—I及广东人参考值的测定

OBJECTIVE: To explore a method for quantitative detection of soluble human leukocyte antigen class I(HLA-I) antigens, with the assistance of which we attempt to define the reference value of the antigen in the population of Guangdong Province. METHODS: Sandwich enzyme-linked immunosorbent assay (ELISA) was employed with W6/32 monoclonal antibody serving as the solid phase antibody and beta2 microglobulin antibody as the first antibody. HRP-labeled second antibody and the substrate were added for enzyme-catalyzed coloring. On the basis of the standard curve obtained by sHLA-I standard reagent in serial dilution, the concentration of sHLA-I antigens in the samples of 60 normal subjects from the population of Guangdong province was detected. RESULTS: The sensitivity of this assay was 2.84 ng/ml with variation coefficients of 5.80% within the same batch and 9.00% between batches. The recovery rate ranged from 98.57% to 100.15%. The level of sHLA-I antigens in normal individuals was 699.54+/-360.10 ng/ml. CONCLUSION: Sandwich ELISA is sensitive, specific and stable in the detection of sHLA-I.     

补阳还五汤对大鼠脑皮层神经元生长的影响

目的观察含补阳还五汤的药物血清对体外培养大鼠脑皮层神经元生长的影响,为补阳还五汤治疗缺血性脑损伤提供实验依据。方法以体外培养新生SD乳鼠大脑皮层神经元为观察对象,采用四唑盐(MTT)比色法测定细胞生长情况。观察含药血清对常规培养及在缺氧状态下神经元生长的影响。结果补阳还五汤对常规培养及在缺氧条件下培养的神经元有显著促进生长作用,与空白血清组比较有显著差(P<0.05)。结论大鼠以补阳还五汤灌胃给药后,其血清中的药物成份有促进体外培养神经元细胞生长的作用。     

异丙酚静脉全麻复合硬膜外腔阻滞麻醉期间CO2气腹对血流动力学和氧耗的影响

OBJECTIVE: To investigate the effects of intraperitoneal CO2 insufflation on the hemodynamics, oxygen consumption (VO2) and carbon dioxide production (VCO2) during intravenous anesthesia with propofol in combination with epidural block. METHODS: Intratracheal intubation was performed after rapid induction of anesthesia and mechanical ventilation was given. Maintenance of anesthesia was achieved using continuous intravenous propofol infusion (2 mg/kg/h) ?N2O inhalation and intermittent epidural administration. Indices of hemodynamics and respiratory function were collected 5 min before induction, 1 min before CO2 insufflation, and 5, 10, 20, 30, 40, 50, 60 min after the start of insufflation and 5 min after the termination of insufflation. RESULTS: The mean arterial pressure (MAP), heart rate (HR), end-tidal PCO2 (P(ET)CO2), VO2 and VCO2 1 min before insufflation were markedly reduced(P<0.01), compared with those recorded before induction. MAP and HR did not undergo any conspicuous changes during CO2 insufflation and 5 min after insufflation termination. Compared with that 1 min before insufflation, PETCO2 was significantly increased 20 min after the start of insufflation (P<0.01), and subsequently carried on the increase though of a lesser scale. VO2 and VCO2 gradually rose after the start of insufflation, and VO2 presented a significantly elevation (P<0.01) 10 min after the insufflation while VCO2 did not show this marked increase(P<0.05) till 20 min after the insufflation in comparison with the levels before insufflation. Subsequently, VO2 continued to rise and VCO2 also retained the increase but of smaller magnitude. CONCLUSION: Intravenous propofol anesthesia combined with epidural block assisted by well-managed excessive ventilation before insufflation can alleviate the adverse effects of CO2 insufflation on respiratory and circulatory systems.     

染色内镜和放大内镜技术是提高早期大肠癌诊治水平的重要手段

应用常规内镜技术难以发现大肠平坦型病变和凹陷型病变。近年来染色内镜和放大内镜技术已经发展成熟,在国外已获广泛应用,可以发现大肠微小病变和早期大肠癌。应用腺管开口分型方法可以预测肿瘤病变的组织学类型及肿瘤的浸润深度,据此可确定行内镜下粘膜剥离术或分片粘膜剥离术将肿瘤切除,抑或行外科手术治疗。在当前我国的胃肠内镜医疗界,应广泛开展染色内镜和放大内镜的临床应用,以早期发现大肠病变,提高我国大肠癌的内镜诊治水平。     

葡聚糖硫酸钠致溃疡性结肠炎小鼠模型的建立

目的　建立小鼠溃疡性结肠炎模型。方法　给小鼠自由饮用5％葡聚糖硫酸钠（DSS）溶液7d后,改为蒸馏水自由饮用10d,如此进行4个循环,每天观察症状。在第1个循环和第4个循环后剖杀小鼠,取出整段结肠行实体显微镜观察及切片组织学研究。结果　所观察的症状、实体显微镜像、组织学病理改变均与人类溃疡性结肠炎类似。结论　此模型制作方法简便,重复性良好,可应用于多种实验研究。     

创伤后肝细胞凋亡与坏死的检测及其意义

目的 探讨严重创伤后肝细胞凋亡及坏死在急性肝功能障碍发病机制中的作用。方法复制多发性骨折合并休克的大鼠创伤模型,采用 Annexin－V－Flous、碘化丙锭(propidium iodid,PI)双标法经流式细胞仪检测创伤后各时间点肝细胞凋亡与坏死的数量变化,结合光镜、电镜和电泳观察细胞凋亡与坏死,并与肝功能变化相比较。结果创伤后早期肝细胞即发生凋亡和坏死,坏死肝细胞的数量进行性升高,与肝功能变化显著呈正相关;凋亡肝细胞在创伤后3h达高峰,部分凋亡肝细胞发生继发性坏死,其数量与肝功能变化显著正相关。结论肝细胞坏死与凋亡是严重创伤后肝功能损害的重要原因,坏死肝细胞是肝功能损害的直接因素,凋亡肝细胞通过发生继发性坏死加重肝功能损害。     

坏宜性脓皮病12例临床分析

报告12例疸性腔皮病患者的性别与年龄构成、并发症、实验室检查、治疗经过和结果，对该病的病因、治疗和预后进行了讨论。     

成人成骨细胞与珊瑚羟基磷灰石的体外生物相容性

目的：研究成人骨髓成骨细胞与珊瑚羟基磷灰石（carolline hydroxyapatite,CHA)在体外培养条件下的生物相容性。方法：抽取健康成人骨髓组织，置于含体积分数为10％胎牛血清的DMEM培养基中培养，传代后改用含地塞米松、β-甘油磷酸钠和维生素C的条件培养基培养，分为CHA复合细胞组和单纯细胞组，不同时间用倒置相关显微镜，HE染色光镜及扫描电镜观察，MTT法进行细胞增殖测定，并进行细胞微量蛋白含量和碱性磷酸酶的定量检测，结果：成人骨髓成骨细胞体外培养时复合或不复合CHA均生长良好，表现出典型成骨细胞的形态特征和生物学特性，CHA利于细胞的贴附，生长与增殖，并对细胞的功能无不良影响，结论：CHA是较理想的骨组织工程支架材料，成骨细胞复合CHA用于骨缺损的修复，具有广阔的临床应用前景。