Supplementation Effect of Early Pregnancy Factor-Positive Serum into Bovine In Vitro Fertilization Culture Medium

PROBLEM: Early pregnancy factor (EPF) has been detected in pregnant bovine serum, and its activity appeared from 24 to 48 hr after insemination. However, in bovine in vitro fertilization (IVF), an EPF-like substance(s) had been detected in the culture medium of fertilized ovum. Therefore, we think that EPF and EPF-like substance(s) are very important materials for the development of the embryo. In this study, we examined the development of the embryo when fertilized bovine ova were cultured with IVF culture medium supplemented with EPF-positive or -negative serum. METHOD OF STUDY: EPF activity of each serum (fetal calf serum [FCS], calf serum [CS], estrus bovine serum, and pregnant bovine serum) was assessed by the bovine-rosette inhibition test. The sera were supplemented in TCM-199 culture medium, and IVF bovine ova were cultured with the media for 6 or 7 days, respectively. The culture media of each group were evaluated for EPF activity by the bovine-rosette inhibition test 48 hr after IVF. The cleavage rate of each group was calculated at 48 hr, and 6 or 7 days after IVF. The culture medium of cumulus cells was also tested for EPF activity. RESULTS: Only the pregnant bovine sera were EPF positive. All the culture media 48 hr after IVF became EPF positive. Additionally, the culture medium of cumulus cells did not have EPF activity. There was no significant difference in the cleavage rate of the EPF-positive and - negative sera 48 hr after IVF. However, the cleavage rate of EPF-positive sera tended to be higher than the negative sera. In contrast, the blastocyst development rates of EPF-positive sera were significantly higher than those of the negative sera 6 to 7 days after IVF (P < 0.05). CONCLUSIONS: The data suggest that an EPF-like substance(s) may be secreted from the in vitro fertilized bovine ovum but not from the cumulus cell, and that the EPF in the pregnant serum may accelerate the development of the bovine embryo in IVF.     

IgA rheumatoid factor in mucosal fluids and serum of patients with rheumatoid arthritis: immunological aspects and clinical significance.

In order to gain insight into the production and clinical significance of IgA rheumatoid factor (IgA-RF) in mucosal fluids of patients with rheumatoid arthritis (RA), we examined tear fluid, saliva and serum from 80 patients with RA. Significant correlations were found between IgA-RF levels in tear fluid and saliva (P=0.002, r=0.57), saliva and serum (P<0.001, r= 0.79), and serum and tear fluid (P<0.001, r=0.31). No significant correlations were found between total IgA levels in these fluids. Comparison between circulating and mucosal IgA-RF levels after correction for total IgA, revealed that mucosal IgA-RF levels are on average 2.5 times higher than circulating IgA-RF levels. Analysis of IgA-RF specificity showed that lacrimal and salivary IgA-RF reactivity with various IgG subclasses is similar and differs from serum IgA-RF specificity. These results indicate local production of IgA-RF in salivary and lacrimal glands and support the view of a common origin of IgA-RF producing B cells present in mucosal tissues. Mucosal and circulating levels of IgA and IgA-RF were not associated with tests that quantify tear fluid production. This indicates that mucosal and circulating levels of IgA and IgA-RF in patients with RA cannot be regarded as markers for the development of secondary Sjögren's syndrome.     

Over-expression of tenascin-C in malignant pleural mesothelioma

Aims:  Tenascin-C is an extracellular matrix glycoprotein known to have anti-adhesive characteristics and to be expressed in various human malignant neoplasms. We hypothesized that the expression of tenascin-C would be increased in human malignant pleural mesothelioma, and its accumulation associated with the prognosis of the patients with this disease.
Methods and results:  Thirty-seven cases of mesothelioma were studied by immunohistochemically using a monoclonal antibody against tenascin-C, and with a semiquantitative scoring system for tenascin-C in different areas of the tumours. In 10 selected cases tenascin-C mRNA in-situ hybridization was also analysed. Since transforming growth factor-beta (TGF-β) is known to induce both the synthesis of tenascin-C and the growth of mesotheliomas, an immunohistochemical analysis of TGF-β1, -β2 and -β3 was also performed. Normal pleura ( n  = 7) and metastatic pleural adenocarcinomas ( n  = 7) were used as controls. Tenascin-C protein was expressed in every histological subtype of malignant mesothelioma, being most prominent in the fibrotic stroma of a tumour, around tumour cells and at the invasive border, whereas tenascin-C mRNA was scarce in tumour cells. The patients with less immunohistochemical expression for tenascin-C tended to live longer ( P  = 0.028 by Fishers' exact probability test). All mesotheliomas showed positivity for at least one isoform of TGF-β.
Conclusions:  In conclusion, high expression of tenascin-C protein in malignant pleural mesotheliomas may play a role in its invasive growth, and might serve as a prognostic marker of the disease.     

c-erbB-2、VEGF和组织蛋白酶D在胃癌中的表达及其相关性

目的：探讨癌基因c-erbB-2、血管内皮生长因子（VEGF）和组织蛋白酶D（Cath-D)在胃癌中的表达及其与胃癌生物学行为的关系。方法：采用免疫组织化学（S－P）法，检测了102例胃癌手术标本中c-erbB-2、VEGF及Cath-D的表达，同时观察了网状纤维分布与c-erbB-2、Cath-D之间的关系，并将检测结果与跟踪随访资料进行了综合分析。结果：102例胃癌组织中c-erbB-2表达阳性率为38.24%（39／102），与胃癌浸润深度（P＜0.05）、淋巴结转移（P＜0.05）密切相关；VEGF表达阳性率为50％（51／102），与胃癌浸润深度（P＜0.01）、生长方式（P＜0.01）、淋巴结转移（P＜0.01）密切相关；Cath-D表达阳性率为81.37%(83/102)，与胃癌浸润深度（P＜0.05）、生长方式（P＜0.05）、淋巴结转移（P＜0.05）及脉管内有无癌栓（P＜0.05）有关。对生存期分析显示：Cath-D、VEGF、c-erbB-2阳性患者预后差，5年生存率低于阴性表达患者。结论：c-erbB-2、VEGF及Cath-D与胃癌的生长、浸润、转移、预后有密切关系，可作为判断胃癌生物学行为和预后的重要指标。     

p38蛋白激酶对大鼠肺泡巨噬细胞活化机制的调控

目的:探讨p38蛋白激酶对LPS诱导大鼠肺泡巨噬细胞激活机制的作用。方法:提取细胞核蛋白,采用Western印迹分析p38蛋白激酶水平。用放射免疫法检测细胞上清TNF-α、IL-8的含量。结果:LPS显著增加肺泡巨噬细胞TNF-α、IL-8的合成,呈剂量依赖性诱导p38的活化。特异性p38蛋白激酶抑制剂SB203580能显著降低LPS诱导的肺泡巨噬细胞核蛋白p38的含量及细胞上清TNF-α、IL-8水平。结论:LPS刺激肺泡巨噬细胞释放炎性细胞因子TNF-α、IL-8受p38蛋白激酶调控。     

女性经前期复发性阿弗他溃疡患者血清IL-6、IL-8、TNF-α和sICAM-1的含量变化及其意义

目的:研究女性经前期复发性阿弗他溃疡(经前期RAU)患者血清中白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、肿瘤坏死因子-α(TNF-α)和可溶性细胞间粘附分子-1(sICAM-1)的变化状况, 并探讨经前期RAU的发病机制。方法:用酶联免疫吸附双抗体夹心法(ELISA)检测女性经前期RAU患者外周血中IL-6、IL-8、TNF-α、sICAM-1的水平, 与正常对照组和无经前发作规律的女性RAU对照组相比。结果:经前期RAU患者的血清IL-6、IL-8、TNF-α水平不仅显著高于正常对照组(P＜0.01), 而且高于女性RAU对照组(P＜0.01)。而sICAM-1无明显变化。女性RAU患者血清中TNF-α亦高于正常对照组(P＜0.05)。结论:经前期RAU患者体内存在IL-6、IL-8、TNF-α介导的免疫功能异常, 可能对RAU局部损害起作用。     

VEGF与血液学恶性疾病

VEGF作为一种血管新生的主要病理生理性调节因子，触发了白血病细胞及多发性骨髓瘤细胞的生长、存活及移动，在造血过程中起重要作用。VEGF的表达及其信号转导，对血液学恶性疾病，尤其是对多发性骨髓瘤的发病机制及临床特性都有重要作用。针对VEGF及其受体直接或间接的治疗，有可能为临床提供一种新的有效的治疗方法。     

神经再生素促大鼠海马神经元生长的研究

目的 研究牛膝提取物神经再生素(NRF)对体外培养的大鼠海马神经元生长的促进作用及其对生长相关蛋白-43(GAP-43)表达的影响.方法 以体外原代培养的胎鼠海马神经元为研究模型,通过相差显微镜采用Scion软件测量不同浓度NRF(0.25mg/L、0.5mg/L、1mg/L)、不同作用时间(6h、12h、24h)对海马神经元神经突起生长的影响;通过实时荧光定量PER观察NRF不同浓度(0.25mg/L、0.5mg/L、1.0mg/L)及不同作用时间(6h、12h、24h)对海马神经元GAP-43基因表达的影响;采用免疫荧光细胞化学法和Western blotting,观察不同浓度NRF(0.25mg/L、0.5mg/L、1.0mg/L)加药24h后,对海马神经元GAP-43表达的影响.结果 NRF可有效地促进海马神经元的生长,加药后24h作用浓度为1mg/L时,作用最强;实时荧光定量RT-PCR、免疫荧光细胞化学法和Western blotting的结果提示,NRF能增加体外培养的海马神经元GAP-43的表达,浓度为1.0mg/L时,作用最佳.结论 NRF能促进体外培养的海马神经元神经突起的生长和GAP-43基因的表达,表明NRF对海马神经元具有神经营养作用.