真核表达人呼吸道合胞病毒F蛋白的纯化方法

目的 真核表达人呼吸道合胞病毒(human respiratory syncytial virus,SV)融合蛋白(fusion protein,),并完成蛋白纯化及纯度测定.方法 根据编码F蛋白的基因序列设计引物,CR方法扩增出3'端带His标签的F基因序列,克隆入pGEM-T-easy载体,经核酸序列分析后,进一步克隆到pcDNA3.1( )真核表达载体,限制性内切酶鉴定,用脂质体Lipofectamine2000转染COS-7细胞,2 h后再用Westem blot检测目的蛋白的表达.Ni柱亲和层析纯化COS-7细胞表达的F蛋白,高效毛细管电泳分析纯化后蛋白纯度.结果 核酸序列分析证实获得带His标签的RSV F基因序列,没有发生无义突变.转染COS-7细胞后,利用Western blot方法检测到F蛋白的特异性条带,纯度达99%以上.结论 初步建立了真核表达RSV F蛋白的纯化方法,为进一步优化RSV F蛋白制备条件及单克隆抗体及诊断试剂等研究奠定了基础.     

Endoscopic closure of fetal membrane defects: comparing iatrogenic versus spontaneous rupture cases.

OBJECTIVE: Currently, physicians manage preterm premature rupture of membranes (PPROM) by expectant management or termination of the gestation. A therapy aimed at sealing membranes would be optimal to maintain the pregnancy and achieve a normal neonate. Our objective was to compare an endoscopic technique for intrauterine closure of fetal membrane defects after both iatrogenic and spontaneous rupture of membranes. METHODS: Our technique was performed on four patients experiencing PPROM spontaneously and four patients after genetic amniocentesis. Intrauterine endoscopy allowed direct visualization of membrane defects. Rapid sequential injections of platelets, fibrin glue and powdered collagen slurry were administered at the site of the defect and of trocar placement. Sonography for amniotic fluid index, nitrazine and fern testing and pad count were performed after each procedure at three intervals: immediately post-procedure, and after 24 and 48 h. RESULTS: Eight patients underwent endoscopic intrauterine sealing of ruptured membranes between 16 and 24 weeks of gestation: four were spontaneous ruptures and four were ruptures post-amniocentesis. In the post-amniocentesis group, three patients delivered viable infants at 26, 32 and 34 weeks. In one patient, the membranes ruptured again 12 h after the sealing procedure and she decided to undergo termination of pregnancy. Of the four spontaneous rupture patients, two experienced preterm labor and delivery within 2 days of the procedure. One patient was diagnosed with fetal demise 12 h post-procedure, and one patient delivered a neonate at 31 weeks of gestation with severe respiratory distress syndrome. CONCLUSIONS: This technique for sealing ruptured membranes is effective after amniocentesis, but may not be of benefit with spontaneous rupture.     

Three and four-dimensional ultrasound in obstetrics and gynecology

PURPOSE OF REVIEW: Developments in ultrasound in general, but even more so in three-dimensional ultrasound, parallel the growth in computing power and speed of computer technology. It is not surprising, therefore, that three-dimensional ultrasound technology is constantly evolving at a fast pace. The purpose of this article is to provide enhanced diagnostic capabilities for the obstetrical and gynecologic provider. RECENT FINDINGS: The most recent advances in three-dimensional ultrasound have to do with two main features. First, an increasingly fast acquisition speed, enabling quick sequences of fast moving organs such as the heart to be captured. Second, the increasing number of different display modalities, making understanding and analysis of normal anatomy and pathology easier for clinicians. SUMMARY: This article highlights a selected number of clinical situations in which three-dimensional ultrasound meaningfully enhances the contribution of this fast evolving diagnostic imaging tool.     

上皮样血管瘤16例临床与病理分析

 目的:探讨上皮样血管瘤（EH）的临床及组织病理学特征。 方法:回顾性分析16例已确诊患者的临床及病理资料,并对相关文献进行综述。 结果:16例患者中女8例,男8例;平均年龄（45.44±12.52）岁;病程3个月~20年;皮损主要表现为红色至暗红色丘疹、结节,可伴瘙痒和糜烂,好发于头皮、耳部,其中发生在头部10例、耳部5例、外阴1例;皮损单发者5例,多发者11例。皮损组织病理学检查均具有典型的血管增生,管壁增厚,内皮细胞呈上皮样突向管腔,管周大量淋巴细胞及嗜酸性粒细胞浸润。结论：上皮样血管瘤少见,诊断需要临床与病理密切结合。加强对该病的认识,可避免误诊误治。     

Transvaginal sonographic characterization of ovarian disease: evaluation of a new scoring system to predict ovarian malignancy

With the advent of high-frequency transvaginal ultrasonography, new opportunities are presented to better define ovarian lesions. The goal of this study was to develop a scoring system using transvaginal sonographic characterization of pelvic/ovarian lesions. Our purpose was to maximize the discrimination between benign and malignant entities. Transvaginal sonographic pelvic images of 143 patients were correlated with surgical findings or histopathology. Of 281 ovaries, 108 had benign lesions (30 endometriomas, 24 teratomas, 21 simple cysts, and 33 other abnormalities) and 20 had malignancies. The scoring system devised was useful in distinguishing benign from malignant masses, with a specificity of 83%, sensitivity of 100%, and positive and negative predictive values of 37 and 100%, respectively. Further experience and refinements of this method of scoring should maximize the benefit of high-resolution transvaginal sonography of ovarian lesions.     

Effect of exogenous surfactant on total respiratory system compliance

We measured total respiratory system compliance (Crs) before and after instilling 25 mg artificial surfactant in 1 ml saline down the endotracheal tube of preterm babies requiring resuscitation at birth, and compared results with data from 6 similar babies receiving saline only. Surfactant did not produce a significant improvement in Crs.     

Real-time spectral analysis of the fetal EEG: a new approach to monitoring sleep states and fetal condition during labor

Adverse perinatal events affecting cerebral functions are a major cause of neonatal mortality, morbidity, and long-term neurologic deficit. Intrapartum fetal EEG, which records fetal brain electrical activity, provides a monitoring modality for evaluating the fetal CNS during labor. In this study, we describe a new approach to such monitoring that is based on real-time spectral analysis of the fetal EEG during labor. Fourteen pregnant women with uncomplicated term pregnancies who went into labor participated in the study. Two suction-cup electrodes were applied to the fetal scalp at the occipitoparietal or parietal region after rupture of membranes. Real-time spectral analysis was used to determine the frequency and amplitude of the fetal EEG signal. The spectral edge frequency (SEF) was calculated as the frequency below which 90% of the power in the power spectrum resides. The average EEG amplitude and the SEF were displayed using the density spectral array technique. Fetal heart rate and intrauterine pressure were also measured. Two fundamental EEG patterns were identified: high-voltage slow activity and low-voltage fast activity. The SEF was found to be an excellent index of cyclic EEG activity. Fetal heart rate demonstrated increased variability and an elevated baseline during low-voltage fast activity, whereas both parameters decreased during high-voltage slow activity. During episodes of variable decelerations in the fetal heart rate, a decrease in the SEF was observed, accompanied by an increased EEG voltage. The results obtained substantiate the presence of sleep cycles in the human fetus. This kind of cortical activity monitoring may enable rapid alertness to cerebral hypoxia and allow for prompt intervention, thereby decreasing the risk for birth asphyxia and subsequent brain damage.