结核病的体液免疫诊断研究(V.PPA—ELISA检测胸、腹水结核抗体)

以PPD为抗原,建立了检测人体胸、腹水中抗结核菌抗体的PPA-ELISA,对方法的特异性、重复性和最适检测条件进行了探讨。63份患者胸、腹水标本的检测结果显示,28例结核性胸膜炎(腹膜炎)患者胸、腹水的特异性抗体水平非常明显地高于35例对照组患者(P<0.001)。本法简便快速且灵敏,可作为结核性胸膜炎(腹膜炎)的辅助诊断方法;其动态观察还可为疗效考核提供依据,作者等建议加以推广应用。     

酶法、离子选择电极法检测血清钙方法的比较

目的用酶法、离子选择电极法(ISE)进行同步检测血清钙的含量,从中找出一种更为简便、灵敏、经济适用、快速准确的检测方法。方法随机抽取门诊病人标本30份,分别用酶法、离子选择电极法检测血清钙的含量,进行统计学处理和分析。结果30份标本中,酶法和离子选择电极法检测发现它们的回收率、线性范围、精密度均符合要求,重复性良好,结果无显著性差异。结论离子选择电极法较酶法速度快,适于抢救急诊重危病人使用;但对实验室条件及外电电源要求较高,电极需定期精心维护保养。酶法较为稳定,适合全自动化仪器和大批量标本检测的应用,但对黄疸较深和乳糜血、溶血等标本结果影响较大。     

VEGF受体3亲和肽亲和特性研究

目的通过测定人源VEGF受体3(VEGFR3)亲和肽(WHGSLKQNLWWY)及VEGF-D与VEGFR3的功能性亲和常数,鉴定亲和肽的亲和特性.方法通过戊二醛耦联法包被多肽,确定最佳多肽包被浓度,最佳多肽与VEGFR3结合反应时间以及包被系数后,采用非竞争性ELISA固项法,得到多肽及VEGF-D与VEGFR3的结合反应曲线,计算多肽及VEGF-D与VEGFR3的亲和常数.结果多肽及VEGF-D与VEGFR3的功能性亲和常数分别为(1.42±1.45)×107mol/L-1和(1.03±1.16)×108mol/L-1.结论该多肽与VEGFR3亲合能力较强,为今后开发以该多肽为基础的靶向治疗药物提供了理论依据.     

重组E.coli. LLO/OVA明显增强小鼠CD11c细胞活性及抗肿瘤免疫

目的 探讨重组E.coli. LLO/OVA 诱导C57BL/6小鼠机体免疫的途径,观察其免疫后小鼠机体抑制B16-OVA黑色素瘤的效果.方法 磁珠分离E.coli. LLO/OVA及E.coli. OVA免疫后小鼠脾脏CD11c、CD4 和CD8 T细胞并检测CD11c细胞诱导同源CD4 和CD8 T增殖水平和细胞因子分泌程度;流式细胞检测小鼠脾脏内肿瘤抗原OVA257-264 SIINFEKL特异的细胞毒T细胞含量.比较两种E.coli. 免疫后,恶性黑色素瘤B16-OVA在小鼠肺内形成瘤结节的数量.结果 与E.coli. OVA相比, E.coli. LLO/OVA免疫后小鼠脾脏CD11c细胞诱导同源CD4 T细胞增殖作用增强、IL-2分泌增高;同时诱导CD8 T细胞增殖和IFN-γ分泌的作用也明显增强;OVA257-264 SIINFEKL特异的CD8 T细胞含量也明显增高;小鼠肺内形成B16-OVA瘤结节平均数明显减少. 结论 E.coli. LLO/OVA有效地诱导小鼠CD11c细胞活化,增强其对CD4 T细胞增殖和IL-2分泌以及对CD8 T细胞增殖、IFN-γ分泌的作用,诱导了更多的OVA特异的CD8 T细胞,使机体产生了更强的抗肿瘤免疫.     

促甲状腺素实验检测进展

介绍新的更灵敏的TSH(促甲状腺素)检测方法的发展及临床意义,强调实验室应独立评价并定期核查灵敏度的方法学指标以评估不同方法,提高对临床的指导作用。     

Repeated vitrification/warming of human sperm gives better results than repeated slow programmable freezing

In this study, we compared the effects of repeated freezing/thawing of human sperm by our in-house method of rapid freezing with slow programmable freezing. Sperm samples from 11 normozoospermic subjects were processed through density gradients and divided into three aliquots: non-frozen, rapid freezing and slow programmable freezing. Sperm in the rapid freezing group had better motility and viability than those in the slow freezing group (P<0.01) after the first, second and third cycles of freezing/thawing, but there was no difference in morphology. In the second experiment, rapid freezing was repeated three times in 20 subjects. The samples from each thawing cycle were evaluated for DNA fragmentation using the alkaline comet assay. DNA fragmentation began to increase considerably after the second cycle of freezing/thawing, but to a level that was not clinically important. In the third experiment, rapid freezing was done repeatedly in 10 subjects, until no motile sperm were observed after thawing. The median number of repeated freezing/thawing that yielded no motile sperm was seven (range: 5–8, mean: 6.8). In conclusion, we demonstrated that repeated freezing/thawing of processed semen using our rapid freezing method gave better results than standard slow programmable freezing. This method can help maximize the usage of precious cryopreserved sperm samples in assisted reproduction technology.     

Bioassay and Radioimmunoassay of Plasma Gastrin in a Case of Zollinger-Ellison Syndrome

Fasting plasma gastrin levels were measured in a patient with the Zollinger-Ellison syndrome by both bioassay and radioimmunoassay. The level remained constant following total gastrectomy, but fell sharply following parathyroidectomy. Although the bioassay gave gastrin levels which were consistently 2 to 3 times higher than those obtained by radioimmunoassay, the values showed a similar trend. However, both methods gave almost identical results for the gastrin contents of antral mucosa and pancreatic tumour samples from the patient. It is suggested that biologically active gastrin-like substances not detectable by radioimmunoassay were present in the patient's plasma.     

Antioxidant and Anticlastogenic Capacity of Prickly Pear Juice

Plants belonging to the genus Opuntia spp. are the most abundant of the Cactaceae family, grown throughout America and the Mediterranean central area. Its fruit, known as cactus pear or prickly pear, is an oval berry grouped in different colors. Some studies have shown its antioxidant activities which may help in preventing chronic pathologies such as diabetes. The purpose of the study was to evaluate the antioxidant capacity of three varieties of prickly pear juice (red-purple, white-green and yellow-orange) in five different concentrations (100, 250, 500, 750, and 1000 mg/mL) by DPPH (1,1-diphenyl-2-picrylhydrazyl radical) colorimetric method, selecting the best variety to determine its anticlastogenic potential against methyl methanesulfonate (MMS). The results indicate that the highest antioxidant was found in the juice of the prickly pear red-purple variety (PPRP), in all concentrations. Its anticlastogenic potential was therefore evaluated with a micronucleus assay. The experiment was run over two weeks. A negative control was included along with a positive control with MMS (40 mg/kg), a group of mice treated with PPRP (25 mL/kg), and three groups with PPRP (in doses of 25, 16.5 and 8.3 mL/kg) plus the mutagen. The PPRP was administered daily by oral gavage and the MMS was injected intraperitoneally five days prior to the end of the experiment. Blood samples were obtained at 0, 24, 48, 72 and 96 h in order to determine the frequency of micronucleated polychromatic erythrocytes (MNPE). The results indicated that PPRP is not a genotoxic agent, on the contrary, it may reduce the number of MNPE. In this regard, the PPRP showed an anticlastogenic effect directly proportional to its concentrations. Thus, the highest protection was obtained with a concentration of 25 mL/kg after 48 h of treatment.