Reactivity of human sera in a sensitive, high-throughput pseudovirus-based papillomavirus neutralization assay for HPV16 and HPV18

Sensitive high-throughput neutralization assays, based upon pseudoviruses carrying a secreted alkaline phosphatase (SEAP) reporter gene, were developed and validated for human papillomavirus (HPV)16, HPV18, and bovine papillomavirus 1 (BPV1). SEAP pseudoviruses were produced by transient transfection of codon-modified papillomavirus structural genes into an SV40 T antigen expressing line derived from 293 cells, yielding sufficient pseudovirus from one flask for thousands of titrations. In a 96-well plate format, in this initial characterization, the assay was reproducible and appears to be as sensitive as, but more specific than, a standard papillomavirus-like particle (VLP)-based enzyme-linked immunosorbent assay (ELISA). The neutralization assay detected type-specific HPV16 or HPV18 neutralizing antibodies (titers of 160-10240) in sera of the majority of a group of women infected with the corresponding HPV type, but not in virgin women. Sera from HPV16 VLP vaccinees had high anti-HPV16 neutralizing titers (mean: 45000; range: 5120-163840), but no anti-HPV18 neutralizing activity. The SEAP pseudovirus-based neutralization assay should be a practical method for quantifying potentially protective antibody responses in HPV natural history and prophylactic vaccine studies.     

Efficacy of intracytoplasmic sperm injection using intentionally cryopreserved epididymal spermatozoa

Microsurgical epididymal sperm aspiration was a great advancein the therapy of patients with non-recon-structable, obstructiveazoospermia, most notably congenital bilateral absence of thevas deferens. Using conventional in-vitro fertilization, pregnancieswere rarely achieved because the rate of oocyte fertilizationwas extremely poor. However, the use of retrieved spermatozoain conjunction with intracytoplasmic sperm injection (ICSI)has dramatically increased the likelihood of embryo formation.Typically, sperm and oocyte harvesting are performed simultaneously.We have investigated whether frozen-thawed spermatozoa workas well as fresh spermatozoa. When we had concluded from ourown population of patients (groups I and II) that they did,we adopted a policy of aspirating spermatozoa, primarily cryopreservingthem and using them for ICSI at a later date. We found the fertilizationrates of this latter cohort of patients (group III) to be excellent(37% per oocyte), and the ongoing pregnancy rate is quite satisfactory(40 % per couple, 29% per cycle). We offer this approach asan alternative to the traditional scheme because it markedlyeases the burden of partner scheduling on both the couple andthe clinicians involved. In addition, assurance of the availabilityof male partner spermatozoa can be attained prior to beginningovulation induction.     

Externalization and internalization of cardiac endothelin receptors during different phases of sepsis in rat.

Objective. To study the redistribution ofendothelin- 1 (ET- 1 ) receptors in two subcellular organelles, the sarcolemreal membrane and the light vesicle, of rat heart during the progression of sepsis. Methods. Sepsis was induced by cecal ligation and puncture (CLP). ET1 receptor was assayed by using[125I]-ET1 binding. Marker enzyme activities, protein yield, and dry-to-wet weight ratio of cardiac membraneswere measured. Results. Septic rat heart exhibited two distinct phases: an initial hyperdynamic phase( 9h after CLP; early stage of sepsis) followed by a hypodynamic ( 18h after CLP, late stage of sepsis) phase. [125I]-ET1 binding study showed that during early stage of sepsis, the Bmax of ET1 receptors was increased by 30% in sarcolemma but decreased by 19% in light vesicles, while during late stage of sepsis, the Bmax was decreased by 24% in sarcolem-ma but increased by 38% in light vesicles. The total binding of sarcolemma and light vesicles was increased by 25% during early stage of sepsis but decreased by 17% during late stage of sepsis. Conclusions. These data indicated that ET1 receptors in the rat heart were externalized from light vesicles to sarcolemmal membranes during early hyperdynamic phase while internalized from surface membranes to intracellu-lar compartment during late hypodynamic phase of sepsis.     

电话传送心电图监测与常规心电图诊断心肌缺血的应用比较

目的：探讨电话传送心电图检测（Ｔｒａｎｓｔｅｌｅｐｏｍｉｃ Ｅｌｅｃｔｒｏｃａｒｄｉｏｇｒａｐｈｉｃ Ｍｏｎｉｔｏｒｉｎｇ,ＴＴＭ）心肌缺血的诊断价值。方法：同步检测９７例心肌缺血患者的常规心电图（ＥＣＧ）、电话传送心电图（ＴＴＭ）,调整后者的监测电极与ＥＣＧ的电极部位完全相同,判断二者的Ｓ－Ｔ、Ｔ缺血变化。结果：ＥＣＧ发现９７例典型Ｓ－Ｔ、Ｔ心肌缺血患者,ＴＴＭ有９０例相同变化（敏感性９２．５％     

壮族人和汉族人高血压与心肌肥厚关系探讨

目的:通过对壮族人和汉族人原发性高血压与左室心肌肥厚关系的研究,从侧面探讨两个民族的原发性高血压的等位基因是否相同,从而指导临床的治疗用药.方法:壮族组30例,汉族组40例,两组患者均为原发性高血压病人.采用彩色多谱勒超声仪进行心脏检查,测量相关径线并进行比较.结果:两组患者的心超测值及左室心肌肥厚情况无显著性差异.结论:壮族人与汉族人原发性高血压左室心肌肥厚情况无差异,说明两者的等位基因有可能相同.     

二尖瓣脱垂与急性风湿性心脏炎的相关性研究

目的探讨二尖瓣脱垂(MVP)与急性风湿性心脏炎(ARC)的相关性。方法对108例风湿热并ARC的患者行超声心动图检查以观察MVP的发生率。结果超声心动图检查发现MVP76例,发生率为70％,其中前叶脱垂57例(53％),后叶脱垂13例(12％),前后叶均脱垂6例(6％)。与对照组比较有明显的差异(P＜0．01)。ARC所致MVP中74例(97％)并发二尖辫关闭不全(MI),且MVP与MI的严重程度密切相关(P＜0．01)。结论ARC是MVP,尤其是二尖瓣前叶脱垂的重要原因之一,而MVP是ARC所致MI的重要因素。     

妊娠期糖尿病对妊娠结局的影响

目的研究GDM对母婴的影响。方法对112例GDM孕妇和112例健康孕妇的妊娠结局进行分析。采用11配对病例—对照研究,配对的条件是孕妇年龄相近(±2),同年同月同医院分娩。结果GDM孕妇手术分娩率、早产率、妊娠高血压综合征和巨大儿发生率都较对照组高。GDM的及早诊断和及时治疗可使巨大儿发生率降低。与新生儿病率有关的因素是GDM的严重程度、血糖控制情况及巨大儿。结论早期诊断GDM及控制血糖是减少妇婴并发症的关键。     

氧化低密度脂蛋白、低氧和5-HT对血管平滑肌细胞5-HT2A受体及胞内游离钙的影响

目的　为探讨粥样硬化病变动脉对 5 - HT收缩反应增加的机制 ,观察与粥样硬化有关因子氧化低密度脂蛋白 (ox L DL)、低氧、5 - HT对血管平滑肌细胞 5 - HT2 A受体及其基因表达 ,以及细胞 [Ca2 ]i的效应。方法　分别将大鼠主动脉平滑肌细胞单独以 5 0 μg/ m l ox L DL 培养 8h,2 %低氧处理 2 4和 48h,10 μm ol/ L 5 - HT处理30 min,放射配基结合分析测定 5 - HT2 A受体 ;RT- PCR和 Southern杂交检测受体基因的 m RNA;激光共聚焦扫描显微镜测单个细胞 [Ca2 ]i。结果　经 ox L DL、低氧、 5 - HT处理 ,血管平滑肌细胞 5 - HT2 A受体显著上调 (P<0 .0 1) ;ox L DL及低氧处理的受体基因的 m RNA表达增加 ;ox L DL、低氧处理后 ,5 - HT刺激的血管平滑肌细胞[Ca2 ]i升高幅度显著加大 (P<0 .0 5及 P<0 .0 1)。结论　 ox L DL、低氧、5 - HT引起血管平滑肌细胞 5 - HT2 A受体上调及细胞 [Ca2 ]i增加 ,可能是粥样硬化动脉对 5 - HT收缩反应增强的部分原因。     

增生性瘢痕和瘢痕疙瘩组织中Ⅰ、Ⅲ型前胶原及TGF-β_1 mRNA的表达

目的：探讨增生性瘢痕（Ｈ）和瘢痕疙瘩（Ｋ）组织中Ⅰ、Ⅲ型前胶原及ＴＧＦ－β１基因表达改变的相互关系及临床意义。方法：总ＲＮＡ抽提试剂盒抽提总ＲＮＡ,斑点杂交检测Ⅰ、Ⅲ型前胶原及ＴＧＦ－β１ｍＲＮＡ稳态水平的改变。结果：Ｈ和Ｋ组织中ＴＧＦ－β１ｍＲＮＡ（１．１９７±０．２３７,１．２０４±０．２４３）表达均高于正常瘢痕和正常皮肤（０．３２７±０．０８１,０．３３１±０．０７８）,Ｐ＜０．０１;Ｋ选择性地Ⅰ型前胶原ｍＲＮＡ表达增强,而Ｈ组织中Ⅰ、Ⅲ前胶原ｍＲＮＡ表达均增强,导致ＫⅠ、Ⅲ前胶原ｍＲＮＡ比值明显高于Ｈ（８．１６４±０．３００,１．６６６±０．２０１,Ｐ＜０．０１）。结论：Ｋ和Ｈ组织中胶原蛋白基因表达类型及强度不同,提示在Ｋ和Ｈ发生发展中具有不同的分子机理：ＴＧＦ－β１在增生性瘢痕疙瘩的发病机理中具有不同的分子机理;ＴＧＦ－β１在增生性瘢痕和瘢痕疙瘩的发病机理中具有一定作用。