磁分离酶联免疫荧光法(MEIF)检测人血清中胰岛素

目的:采用磁微粒分离酶联免疫荧光(MEIF)分析技术建立一种简便、高敏感性、定量检测人血清胰岛素(insulin)的新方法.方法:选用2株识别人胰岛素不同表位的单克隆抗体(mAb).一株mAb用异硫氰酸荧光素(FITC)标记, 另一株用碱性磷酸酶(AP)标记;偶联羊抗FITC抗体的磁珠用作固相分离载体, 4-甲基磷酸伞型酮用作荧光底物.结果:成功建立了定量检测人血清胰岛素的MEIF, 灵敏度2.0 μIu/mL, 线形范围0～188.52 μIu/mL, 批内变异系数(CV)为4.3%～5.2%, 批间CV为2.6%～9.5%, 稀释回收率为93%～117%, 加标回收率为94%～113%.实际样品的测定结果与倍爱康公司商品化人insulin磁分离发光系统检测试剂盒的检测结果比较, 具有良好的相关性.结论:MEIF定量测定人insulin方法成本低、灵敏度高、稳定性好, 在临床免疫检测领域具有广阔的应用前景.     

Matrix metalloproteinase expression in basal cell carcinoma: relationship between enzyme profile and collagen fragmentation pattern

Matrix metalloproteinases (MMPs) with collagenolytic and gelatinolytic activities are up-regulated in basal cell carcinoma. In the present study we demonstrate that the major collagenolytic enzyme detected is MMP-1 (interstitial collagenase) while gelatinolytic enzymes include both MMP-2 (72-kDa gelatinase A) and MMP-9 (92-kDa gelatinase B). Significant fractions of all three enzymes are present as active forms. In spite of the fact that high levels of gelatinolytic enzymes are present, the major fragmentation products resulting from digestion of intact type I collagen are the 1/4 and 3/4 fragments (products of MMP-1-mediated digestion). Thus, it appears that the gelatinolytic enzymes are not capable of degrading the collagen fragments as rapidly as they are produced. Since previous studies have demonstrated that interaction of interstitial fibroblasts with high molecular weight fragments of type I collagen leads to increased MMP production, the present results suggest a mechanism underlying altered function of stromal elements in the connective tissue adjacent to the growing neoplasm.     

Absence of proteinase-activated receptor-1 signaling affords protection from bleomycin-induced lung inflammation and fibrosis

Activation of the coagulation cascade is commonly observed in the lungs of patients with both acute and chronic inflammatory and fibrotic lung disorders, as well as in animal models of these disorders. The aim of this study was to examine the contribution of the major thrombin receptor, proteinase-activated receptor-1 (PAR-1), during the acute inflammatory and chronic fibrotic phases of lung injury induced by intratracheal instillation of bleomycin in mice. Inflammatory cell recruitment and increases in bronchoalveolar lavage fluid (BALF) protein were attenuated by 56 +/- 10% (P < 0.05) and 53 +/- 12% (P < 0.05), respectively, in PAR-1-deficient (PAR-1-/-) mice compared with wild-type (WT) mice. PAR-1-/- mice were also protected from bleomycin-induced pulmonary fibrosis with total lung collagen accumulation reduced by 59 +/- 5% (P < 0.05). The protection afforded by PAR-1 deficiency was accompanied by significant reductions in pulmonary levels of the potent PAR-1-inducible proinflammatory and profibrotic mediators, monocyte chemoattractant protein-1 (MCP-1), transforming growth factor-beta-1 (TGF-beta1), and connective tissue growth factor/fibroblast-inducible secreted protein-12 (CTGF/FISP12). In addition, PAR-1 was highly expressed in inflammatory and fibroproliferative lesions in lung sections obtained from patients with fibrotic lung disease. These data show for the first time that PAR-1 signaling plays a key role in experimentally induced lung injury, and they further identify PAR-1 as one of the critical receptors involved in orchestrating the interplay between coagulation, inflammation, and remodeling in response to tissue injury.     

腺病毒伴随病毒表达载体表达人乳头瘤病毒18型E6E7基因构建及其转化作用的鉴定

目的：探讨人乳头瘤病毒（HPV）的E6E7基因在细胞恶性转化中所起的作用。方法：将人乳头瘤病毒（HPV）的E6E7基因克隆至腺病毒伴随病毒表达载体中，通过包装的重组病毒感染，将E6E7基因导入并整合到永生293细胞的基因组中。结果：本研究成功地构建了HPV18 E6E7 AAV病毒并感染了永生293细胞，PCR／Southern杂交分析表明E6E7基因在转化细胞293TL中确有表达，转化细胞293TC和293TL具有明显的转化表型，和亲本293细胞相比，生长速度快，接触抑制消失，集落形成率提高20倍，且集落明显增大，形成时间短。结论：成功地构建了HPV18 E6E7 AAV病毒，HPV18 E6E7基因引起永生化人上皮细胞293的恶性转化。此病毒可用于感染正常上皮细胞，研究其致癌机制。     

脑出血后的病理改变与T淋巴细胞反应

目的:探讨脑出血血肿周围局部病理改变及细胞免疫机制。方法:用免疫组化方法及组织HE染色法观察20只大鼠尾壳核脑出血24 h,3天、7天血肿周围区域的组织病变及CD3 、CD8 T淋巴细胞的分布形式和表达时程。结果:①在脑出血后24 h血肿周围即可见明显血管源性水肿,大量炎性细胞浸润,以中性粒细胞为主,少量散在淋巴细胞、小胶质细胞、星形细胞、少突胶质细胞弥漫浸润;出血3天、7天以淋巴细胞、巨噬细胞浸润为主,小胶质细胞、星形细胞、少突胶质细胞增生明显,双侧半球皮层下和血管周围出现小胶质细胞结节;②出血灶周围CD3 和CD8 T淋巴细胞浸润在出血后24 h已可见,出血后3天组和出血后7天组,CD3 和CD8 阳性细胞数明显高于出血后24 h组。结论:①脑出血期以中性粒细胞反应为主,以后以淋巴细胞反应为主,胶质细胞反应在出血后24 h已发生,持续一周以上;②CD3 、CD8 阳性细胞的出现提示它们参与了出血后脑损伤的病理过程,在出血7天内, CD3 和CD8-阳性细胞反应呈增强趋势。     

Prevalence and severity of asthma and allergies in schoolchildren in Lhasa, Tibet

BACKGROUND: The International Study of Asthma and Allergies in Childhood (ISAAC) demonstrated that large variations existed in the prevalence of symptoms of asthma, allergic rhinoconjunctivitis and eczema throughout the world and that environmental factors and lifestyle customs are major determinants of the prevalence and severity of these diseases. However, the relevant data about children living at high-altitude locations were considered to be underreported. OBJECTIVE: The ISAAC Phase III programme was carried out in Lhasa, the Tibetan Autonomous Region in China, at an elevation of 3658 m above sea level to examine the occurrence of asthma, allergic rhinoconjunctivitis and eczema in schoolchildren aged 13-14 years. METHODS: All 3196 schoolchildren in eight public junior high schools in urban Lhasa who were confirmed to be 13-14 years old were invited and participated in both written and video questionnaire investigations, among which 3190 pieces of data (49.8% of boys and 50.2% of girls) were validated and analysed. RESULTS: Among the overall observations, the prevalence of 'having ever experienced wheezing', 'current wheezing' and 'diagnosed to have asthma' was 1.4%, 0.8% and 1.1%, respectively. The prevalence of current exercise-induced asthma and current nocturnal cough was 7.1% and 4.6%, respectively. The current prevalence of allergic rhinoconjunctivitis and eczema was 5.2% and 0.4%, respectively. In addition, the prevalence of rhinoconjunctivitis symptoms during the past 12 months showed no discernable differences throughout the year. CONCLUSION: The prevalence of asthma, allergic rhinoconjunctivitis and eczema over the past 12 months was the lowest among the centres, that performed ISAAC worldwide.     

舒必利与氯丙嗪合用治疗慢性精神分裂症阴性症状的对照研究

目的 探讨舒必利与氯丙嗪及舒必利,氯丙嗪治疗慢性精神分裂症阴性症状的疗效。方法 将60例以阴性症状为主要表现的慢性精神分裂症病人随机分为舒必利合并氯丙嗪组(研究组)及舒必利组,氯丙嗪3个组。用临床疗效评定标准和BPRS量表评定疗效。结果 研究组的显效率(显进 痊愈)明显比氯丙嗪组及舒必利组高,x~2检验P<0.05提示舒必利合并氯丙嗪治疗比单一用药疗效显著。舒必利组氯丙嗪组显效率比较无显著性差异(P<0.05)。结论 舒必利合并氯丙嗪治疗慢性精神分裂症阴性症状的疗效比单一用药更好,副作用更少。     

Fas在妊娠过程中表达变化的研究

目的探讨Fas在正常妊娠过程及妊娠期高血压疾病(HDCP)中的表达变化及意义。方法分别选取正常妊娠早期和晚期及HDCP胎盘绒毛组织。采用免疫组织化学方法检测Fas表达,并用高清晰彩色病理图像分析系统对其表达进行分析。结果Fas在妊娠早期胎盘组织中极少表达,妊娠晚期胎盘组织中表达增高。HDCP胎盘组织中Fas表达明显升高,与前两组比较差异有极显著性(P<0.01)。结论孕早期滋养细胞表面Fas表达在母胎免疫耐受中发挥重要作用,孕晚期Fas表达增加可能是引起HDCP的原因之一。     

妊娠高血压患者血浆中降钙素基因相关肽的测定及其临床意义

本文测定了３０例正常育龄妇女，２８例正常妊娠以及３６例妊娠征患者血浆中降钙素基因相关肽的水平，结果发现，轻度妊高生与正常妊娠相比，血浆中ＣＧＲＰＫ是有下降但不显著（Ｐ〉０．０５）。中重度妊高征与正常妊娠相比，差异有非常显著性，因此推测，妊娠状态下血浆中降钙素基因相关肽的减少可能是妊娠高血压症产生的重要原因之一。