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.     

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。     

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.     

The clinical usefulness of the fingers-to-palm ratio in different hand microcirculatory abnormalities

A non-invasive nuclear medicine technique was developed to screen patients with painful hands so as to separate patients with a normal from those with an abnormal microcirculation of the hands in different clinical conditions. Such a technique is important, as the other methods available are either subjective or rather complicated. The study population consisted of 10 healthy individuals, 23 patients with Raynaud's syndrome and 15 patients with mixed connective tissue disease (MCTD). Sixty gamma-camera images of the hands (1 s each) were recorded after a bolus injection of 99Tcm-DTPA via a dorsal foot vein. Regions of interest were drawn on the summed images around the fingers and the palmar region. The fingers-to-palm ratio was then calculated from the total counts inside these regions of interest separately for each hand. The mean fingers-to-palm ratio was 0.94+/-0.18 (0.71-1.25) for the healthy group, 0.57+/-0.22 (0.21+/-1.11) for the MCTD group and 0.40+/-0.14 (0.18-0.77) for the Raynaud's patients. Analysis of variance showed these differences to be highly significant (P < 0.001). There were also significant differences between 6 MCTD patients in an active (mean 0.48) and nine patients in an inactive (mean 0.66) clinical state (two-sample t-test: P < 0.05). There were no significant differences between the fingers-to-palm ratios of the left and right hands of the same patients (one-sample t-test). Of the 23 primary Raynaud's patients, capillary microscopic data were pathological in only eight (34%). We conclude that our method is able to differentiate between patients with normal and those with abnormal microcirculation of the hands. Although measurement of the fingers-to-palm ratio is not a specific method, it is useful both for staging and in the follow-up of patients.     

Noonan's and DiGeorge syndromes with monosomy 22q11

A boy with the dysmorphic features of Noonan's syndrome and pulmonary valve stenosis who had evidence of hypoparathyroidism and abnormal T lymphocyte numbers in the neonatal period is reported. He had a normal karyotype but molecular analysis revealed a submicroscopic deletion within chromosome 22q11, the region deleted in DiGeorge syndrome. Thus this child has both Noonan's syndrome and DiGeorge syndrome; 22q11 is a candidate region for a gene defective in Noonan's syndrome.     

Elevated level of Helicobacter pylori antibodies in Henoch-Schonlein purpura

INTRODUCTION: Henoch-Sch?nlein purpura is a non-thrombocytopenic haemorrhagic syndrome characterized by deposition of immunocomplexes in the wall of the arterioles, capillaries and venules, involving the skin, joints, gastrointestinal mucosa, and glomeruli. Several causal factors could be responsible for its evaluation, however the exact exciting agent have not been identified yet. The Helicobacter pylori infection is suggested as a possible cause of Henoch-Sch?nlein purpura by some authors too (Gasparini and al: Eur J Gastroenterol, 1997, 9, 231-233). AIM: Between 1995-2000 11 adult patients (64 +/- 10 year) suffering from Henoch-Sch?nlein purpura were admitted in our department taking particular care over detecting Helicobacter pylori infection. METHOD: Patients were classified as having either Henoch-Sch?nlein purpura in acute phase (5) or in remission phase (6). Each patient underwent panendoscopy with 2-2 biopsies being taken from the corpus, antral and duodenal mucosa. It was investigated the type of inflammation and the existence of Helicobacter pylori. The authors studied laboratory and immunological profiles of all the patients. Previously non of the patients got eradication therapy. RESULT: The serological investigations revealed Helicobacter pylori infection in ten investigated patients. Patients in the acute phase had significantly higher level of anti H. pylori IgG (86 +/- 32 versus 32.5 +/- 23 U/ml) (p < 0.05) compared to remission. Anti H. pylori IgA were elevated with significant difference in remission phase (3.09 +/- 1.78 versus 1.96 +/- 0.58 ratio) (p < 00.5). TNF-alpha level were significantly increased in acute phase (58.8 +/- 18 versus 27.3 +/- 5 pg/ml) (p < 0.001). Serum IgA level were also significantly higher in acute phase (5.44 +/- 1.04 versus 3.49 +/- 1.14) (p < 0.05). Anti Streptococcus DN-ase B were higher only in two patients. ANCA were negative in the all patients. CONCLUSION: According to authors findings the results suggest that seropositivity for Helicobacter pylori may be a risk factor for Henoch-Sch?nlein purpura. The causal role of Helicobacter pylori in the developing of Helicobacter pylori induced extraintestinal manifestations is equivocal however, it can not be ruled out with absolute certainty. To better understanding of this entity further clinical and experimental examinations have to be performed.     

From the Cover: An aberrant island-dwelling theropod dinosaur from the Late Cretaceous of Romania

Islands are noted for the occurrence of aberrant, endemic, and dwarfed taxa (the “island effect”). Late Cretaceous vertebrate assemblages of Romania and elsewhere in Europe are classic examples of island faunas in the fossil record, and are characterized by dwarfed herbivorous dinosaurs and other endemic taxa that are noticeably primitive relative to their mainland contemporaries. Fossils of the predators inhabiting the European paleoislands, however, are exceptionally rare and fragmentary. We describe a new dromaeosaurid theropod, based on an articulated skeleton from the Maastrichtian of Romania, which represents the most complete predatory dinosaur from the middle to Late Cretaceous of Europe. This taxon is characterized by a peculiar body plan, most notably extensive fusion in the hand and distal hindlimb, a highly retroverted pelvis with enlarged femoral muscle attachments, and a pair of hyperextensive pedal claws. However, unlike the island-dwelling herbivorous dinosaurs, its closest relatives are contemporary similar-sized Laurasian taxa, indicating faunal connections between Asia and the European islands late into the Cretaceous. This theropod provides support for the aberrant nature of the Late Cretaceous European island-dwelling dinosaurs, but indicates that predators on these islands were not necessarily small, geographically endemic, or primitive.     

Plasma Homocysteine Levels, The Prevalence of Methylenetetrahydrofolate Reductase Gene C677T Polymorphism and Macrovascular Disorders in Systemic Sclerosis: Risk Factors for Accelerated Macrovascular Damage?

The purpose of this study was to investigate plasma homocysteine (Hcy) levels in patients with systemic sclerosis (SSc) and to study the association between plasma Hcy, C677T polymorphism of 5,10-methylenetetrahydrofolate reductase (MTHFR), and the clinical manifestations in SSc. Associations of Hcy level, C677T MTHFR polymorphism, and macrovascular diseases were investigated in 152 patients with SSc and 58 controls. No significant differences in Hcy levels and MTHFR genotypes were found in SSc patients compared to controls or in SSc patients with limited cutaneous compared to diffuse disease. Significantly higher Hcy concentration was observed in patients with macroangiopathy/thromboembolic events compared to patients without such clinical manifestations (p?<?0.05). There was significant correlation between age and macrovascular disorders, between Hcy level and the disease duration (r?=?0.164; p?<?0.05). Seventy-one percent of patients with macrovascular disorders had MTHFR polymorphism. In addition, 45% of patients with hyperhomocysteinemia had pulmonary hypertension. The presence of MTHFR C677T mutation influences the incidence of macrovascular abnormalities in SSc patients. Elevated Hcy levels may be associated with disease duration and the evolution of macrovascular disorders and pulmonary hypertension in SSc.