Cholangiocyte endothelin 1 and transforming growth factor beta1 production in rat experimental hepatopulmonary syndrome

BACKGROUND & AIMS: Hepatic production and release of endothelin 1 plays a central role in experimental hepatopulmonary syndrome after common bile duct ligation by stimulating pulmonary endothelial nitric oxide production. In thioacetamide-induced nonbiliary cirrhosis, hepatic endothelin 1 production and release do not occur, and hepatopulmonary syndrome does not develop. However, the source and regulation of hepatic endothelin 1 after common bile duct ligation are not fully characterized. We evaluated the sources of hepatic endothelin 1 production after common bile duct ligation in relation to thioacetamide cirrhosis and assessed whether transforming growth factor beta1 regulates endothelin 1 production. METHODS: Hepatopulmonary syndrome and hepatic and plasma endothelin 1 levels were evaluated after common bile duct ligation or thioacetamide administration. Cellular sources of endothelin 1 were assessed by immunohistochemistry and laser capture microdissection of cholangiocytes. Transforming growth factor beta1 expression and signaling were assessed by using immunohistochemistry and Western blotting and by evaluating normal rat cholangiocytes. RESULTS: Hepatic and plasma endothelin 1 levels increased and hepatopulmonary syndrome developed only after common bile duct ligation. Hepatic endothelin 1 and transforming growth factor beta1 levels increased over a similar time frame, and cholangiocytes were a major source of each peptide. Transforming growth factor beta1 signaling in cholangiocytes in vivo was evident by increased phosphorylation and nuclear localization of Smad2, and hepatic endothelin 1 levels correlated directly with liver transforming growth factor beta1 and phosphorylated Smad2 levels. Transforming growth factor beta1 also stimulated endothelin 1 promoter activity, expression, and production in normal rat cholangiocytes. CONCLUSIONS: Cholangiocytes are a major source of hepatic endothelin 1 production during the development of hepatopulmonary syndrome after common bile duct ligation, but not in thioacetamide-induced cirrhosis. Transforming growth factor beta1 stimulates cholangiocyte endothelin 1 expression and production. Cholangiocyte-derived endothelin 1 may be an important endocrine mediator of experimental hepatopulmonary syndrome.     

严重急性呼吸综合征冠状病毒刺突蛋白在酿酒酵母中的表达和纯化

目的构建重组表达质粒pYES6-S，诱导SARS冠状病毒刺突蛋白（spikeprotein）在酿酒酵母中的表达并纯化。方法反转录获得SARS冠状病毒DNA片段，PCR法获得刺突蛋白基因互相重叠的四部分片段，酶切连接成全长，在大肠埃希菌E．coliJM109中构建重组表达克隆pYES6-S，在酿酒酵母巾诱导表达并纯化。结果重组克隆pYES6～S经酶切、测序鉴定确定，插入片段大小、方向、碱基匹配与预期一致，获得1个完整的刺突蛋白基因，表达产物纯化后，电泳鉴定，相对分子质量大小近110000。结论 成功克隆SARS冠状病毒刺突蛋白基因全长，并在酿酒酵母中诱导表达成功，有助于抗SARS分子疫苗的研究。     

Prevalence of SENV-H viraemia among healthy subjects and individuals at risk for parenterally transmitted diseases in Germany

summary . The prevalence of a newly described DNA virus (SENV-H) was examined in a population of 599 individuals by polymerase chain reaction (PCR). All individuals were assigned to a nonrisk or a risk group depending on the presence of historical or serological factors indicating an increased risk for parenterally transmitted diseases. In a group of 226 healthy blood donors, 38 (16.8%) were found to be SENV-H viraemic. The highest prevalence of SENV-H viraemia was observed among patients infected by HIV (28 of 63; 44.4%). Contrarily, of 78 individuals on maintenance haemodialysis, only 10 (12.8%) were found positive in the SENV-H PCR. Our results demonstrate that SENV-H viraemia is widespread in the general population. Therefore, it seems to be questionable if parenteral transmission is the main route for spreading SENV-H. The hepatitis-inducing capacity of SENV-H is unclear. However, taking our clinical and epidemiological data into account it seems unlikely that this virus is responsible for hepatitis.     

紫外线减毒日本血吸虫尾蚴免疫小鼠的皮肤组织细胞反应

目的 :探讨紫外线减毒血吸虫尾蚴免疫宿主的皮肤组织在抗攻击感染中的作用。方法 :88只小鼠分为 4组 ,1、2组分别以 50± 3条紫外线减毒日本血吸虫尾蚴免疫或日本血吸虫尾蚴感染 ,5wk后以 10 0 0± 10 0条尾蚴进行攻击感染 ;3、4组分别以减毒尾蚴或尾蚴 10 0 0± 10 0条免疫或初次感染。各组均于感染或接种后 6- 12 0 h定时取局部皮肤组织作病理观察。结果 :( 1)免疫攻击组皮肤组织内炎症细胞杀伤童虫现象最明显、炎症反应最强且持续时间最长 ,嗜酸性粒细胞( EOS)浸润百分数最高 ;( 2 )感染攻击组呈现相似炎症反应但程度略轻 ;( 3)免疫对照组减毒童虫在皮肤内滞留时间延长 ,至 12 0 h皮下仍可见较多童虫 ;( 4 )感染对照组皮肤内的童虫数于 72 h后明显减少 ,童虫周围轻微炎症反应。电镜观察可见免疫攻击组童虫内部结构破坏 ,虫体周围 EOS、淋巴细胞增多 ,肥大细胞颗粒溶解。结论 :提示皮肤组织的细胞免疫反应在血吸虫减毒尾蚴免疫的宿主抗攻击感染保护性免疫中起重要作用。     

联合应用激光微切和实时定量RT-PCR检测单个胰岛的基因表达

在显微镜下用紫外激光微切机将单个胰岛从胰腺切片中切下，提取RNA后逆转录成cDNA并在实时定量RY—PCR中得到有效的扩增，其表达量与细胞数量成正比。     

Stage determination and therapeutic decision in human African trypanosomiasis: value of polymerase chain reaction and immunoglobulin M quantification on the cerebrospinal fluid of sleeping sickness patients in Côte d'Ivoire

In human African trypanosomiasis (HAT), two disease stages are defined: the first, or haemo‐lymphatic stage, and the second, or meningo‐encephalitic stage. Stage determination forms the basis of therapeutic decision and is of prime importance, as the drug used to cure second‐stage patients has considerable side‐effects. However, the tests currently used for stage determination have low sensitivity or specificity. Two new tests for stage determination in the cerebrospinal fluid (CSF) were evaluated on 73 patients diagnosed with HAT in Côte d'Ivoire. The polymerase chain reaction (PCR) detecting trypanosome DNA (PCR/CSF) is an indirect test for trypanosome detection whereas the latex agglutination test detecting immunoglobulin M (LATEX/IgM) is an indicator for neuro‐inflammation. Both tests were compared with classically used tests, double centrifugation and white blood cell count of the CSF. PCR/CSF appeared to be the most sensitive test (96%), and may be of use to improve stage determination. However, its value for therapeutic decision appears limited, as patients whose CSF was positive with PCR were successfully treated with pentamidine. This result confirms those of previous works that showed that some patients with trypanosomes in the CSF could be treated successfully with pentamidine. LATEX/IgM, which depending on the cut‐off, showed lower sensitivity of 76% and 88%, but higher specificity of 83% and 71% for LATEX/IgM 16 and LATEX/IgM 8 respectively, appears more appropriate for therapeutic decision making.     

Acute lung injury due to parvovirus pneumonia

Wardeh A, Marik P, (Department of Critical Care, St. Vincent Hospital, Massachusetts, USA and the University of Massachusetts and Medical Intensive Care Unit St. Vincent Hospital, Massachusetts, USA). Acute lung injury due to parvovirus pneumonia (Case Report). J Intern Med 1998; 244 : 257–60.  

Objective

Human parvovirus B19 is responsible for the common childhood exanthematous illness, erythema infectiosum. Adults infected with B19 have been reported to develop a febrile illness associated with arthritis. Life threatening parvovirus infections in non-immunocompromised adults have not been reported to date. We report a previously healthy middle aged female who developed severe parvovirus pneumonia. The patient recovered with supportive care provided in the ICU. Parvovirus may represent an under diagnosed cause of community acquired pneumonia.     

A cross-sectional study on HGV infection in a rural population

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用PCR诊断不同虫荷量的大鼠卡氏肺孢子虫肺炎的价值

目的　评价在不同虫荷量条件下用 PCR诊断大鼠卡氏肺孢子虫肺炎 (PCP)的价值。　方法　将 PCP大鼠随机分为药物治疗组及未治疗组 ,测定其肺虫荷量 ,收集其支气管灌洗液 (BAL F)和血清标本 ,用 PCR和半套式 (Sn) PCR检测标本中的卡氏肺孢子虫基因 ,比较 PCR和环六亚甲基四胺银 (GMS)染色结果 ,及与病鼠肺虫荷量的关系。　结果　虫荷量高的未治疗组病鼠的 BAL F标本 PCR、Sn PCR及 GMS染色阳性率分别为 10 0 %、10 0 %和 92 .6 % ,差异无显著性 (P>0 .0 5 ) ;而虫荷量低的治疗组的 PCR(88.1% )、Sn PCR(94 .9% )阳性率均显著高于 GMS染色镜检阳性率 (30 .5 % ) (P<0 .0 5 )。肺虫荷量 >5 0 0个 /mg肺的 PCP大鼠 ,血清 PCR阳性率显著高于≤ 5 0 0者 (分别为 4 2 .9%和 1.7% )。　结论　PCR可用于虫株负荷量较低时的 PCP诊断 ,对判断 PCP病情及疗效考核亦有一定的帮助。     

甲巯咪唑治疗儿童甲状腺功能亢进症不良反应的观察

目的 研究甲巯咪唑治疗儿童甲状腺功能亢进症（简称“甲亢”）不良反应发生情况。 方法 回顾性分析中国医科大学附属盛京医院2015年1月—2021年5月住院的304例甲亢患儿的临床资料。统计甲巯咪唑治疗相关不良反应的发生情况,探讨常见不良反应的危险因素。 结果 304例患儿中,出现甲巯咪唑治疗相关不良反应87例（28.6%）,其中男性20例（23%）,女性67例（77%）。较常见的不良反应有粒细胞减少（12.8%）、皮疹（11.8%）、丙氨酸氨基转移酶升高（9.5%）及关节疼痛（3.0%）,部分患儿可同时或间断出现多种不良反应。粒细胞减少主要出现在用药后3个月内（25/39,64%）;丙氨酸氨基转移酶升高主要发生在用药后1个月内（17/29,59%）;皮疹主要发生在用药后3个月内（30/36,83%）。对症治疗后上述不良反应大部分可恢复正常。多因素logistic回归分析显示,年龄小、治疗前中性粒细胞绝对计数低是甲巯咪唑治疗后粒细胞减少的危险因素（P<0.05）。 结论 甲巯咪唑治疗儿童甲亢不良反应较常见,多发生在用药后3个月内,多数经过对症治疗可缓解。年龄越小,中性粒细胞绝对计数基线值越低,越容易发生粒细胞减少。