克山病患者间质性肺炎的病原学探讨

克山病病因未明,患者常有多脏器感染性病变,尤以肺脏明显,其发病特征提示与肠道病毒感染有关,我们采用套式聚合酶链反应探讨肠道病毒感染与/克山病患者间质性肺炎发病的关系。     

克山病患者心肌中肠道病毒Vp1结构蛋白检测

目的 :硒缺乏可能是克山病发生的条件因子 ,而非特异性致病因素。为探讨克山病的发生与肠道病毒感染的关系。方法 :应用柯萨奇B5病毒的Vp1蛋白单克隆抗体 ,采用免疫组化方法 ,对黑龙江、山东、云南 3省区急型、亚急型、慢型、潜在型克山病患者尸检心肌组织共 83例 ,风心病 10例     

应用核酸分子杂交技术检测心肌炎组织中的肠道病毒RNA

用生物素标记ｐＣＢＩＩＩ／３５和ｐＣＢＩＩＩ／５１肠道病毒属特异性探针，对４５心肌炎和心肮病尸检或心肌活检心脏组织进行原位杂交，结果显示：１２例（２６．６％）存在肠道病毒ＲＮＡ。柯萨基Ｂ３病毒型特异性探针ｐＣＢＩＩＩ／２９进一步杂交表明，１２例经证实存在肠道病毒ＲＮＡ的心肌组织中５例（４１．７％）出现阳性杂交信号。阳性杂信号主要位于单个或多个心肌细胞的胞浆和心肌的间质组织，偶见竽心肌小血管内皮细胞     

潜在型克山病的血液流变学变化

克山病患者存在血液流变学指标的改变，在克山病模型中报道较多，在克山病患者中报道较少．本文对宁县于午岭林区克山病重灾区某林场职工进行普查诊断的潜在型克山病患者32例，进行了血液流变学检测。现报告如下：资料和方法1病例选择对宁县子午岭林区克山病重灾区某林场职工进行普查筛选，对象均为长期居住于病区的居民，根据（克山病防治工作标准》执行。全部病例进行了详细体检，心电图及X线检查。共筛选出克山病48例。再进行超声心动图及心功能测定，除外伴有心功能不全者后，共确诊潜在型克山病32例。其中男14例，女18例。年龄30～56…     

5例感染伴随性噬血细胞综合征尸检观察

我们对５例感染伴随性噬血细胞综合征（ｉｎｆｅｃｔｉｏｎａｓｏｃｉａｔｅｄｈｅｍｏｐｈａｇｏｃｙｔｉｃｓｙｎｄｒｏｍｅ，ＩＡＨＳ）的尸检病理组织形态及免疫组化结果进行观察。一、材料和方法标本来自我院病理科１９８０～１９９７年尸检存档蜡块。全部组织常规制...     

不同类型克山病心肌间质纤维化的形态学分析

目的揭示克山病心肌间质中胶原的类型、分布、排列与含量。方法采用本室克山病尸检材料，应用苦味酸－天狼星红染色和偏振光显微镜观察和图像分析技术，系统地研究了克山病心肌胶原的变化。结果在急性克山病的瘢痕灶内，胶原排列成疏网状，Ⅲ型胶原比例增加。慢性克山病的瘢痕中，胶原排列致密，Ⅰ型胶原比例增加，Ⅰ／Ⅲ型胶原比值增大。在心肌细胞非坏死区域胶原含量明显高于对照组，出现间质纤维化，以慢性克山病最明显。亚急性和慢性克山病的心内膜厚度和血管周围胶原体密度明显高于对照组。结论在克山病心肌细胞变性坏死的同时伴随着胶原代谢的紊乱，胶原蓄积，心肌间质胶原发生重构，是克山病心功能不全的重要因素之一。     

冷冻与石蜡肾活检标本中PLA2R1表达的临床诊断意义比较

目的探讨磷脂酶A2受体1(PLA2R1)在肾脏穿刺的冷冻标本及石蜡标本中的表达,分析不同标本中阳性率的临床病理诊断意义。方法收集2013年1月~2019年2月江门市中心医院行肾脏穿刺患者的标本及临床病理资料,其中同时有肾活检冷冻标本及石蜡标本的患者215例,仅有石蜡标本的患者785例。分别利用免疫荧光及免疫组化法检测PLA2R1在肾活检冷冻和石蜡标本中的表达,并分析PLA2R1表达与临床病理特征的相关性。结果在215例冷冻标本中,PLA2R1阳性率为19.1%;在1000例石蜡标本中,PLA2R1阳性率为32.9%,两组中PLA2R1的阳性率差异有统计学意义(P<0.001)。在冷冻和石蜡标本中,PLA2R1表达与患者性别、年龄及临床分期无关,与病理类型相关。在冷冻和石蜡标本中,PLA2R1阳性均集中于特发性膜性肾病(72.0%和79.8%),但非特发性膜性肾病中,则分别为3.0%和19.4%。结论肾活检冷冻及石蜡标本中PLA2R1表达均与特发性膜性肾病相关,但冷冻标本在特发性膜性肾病诊断中更有价值。     

不同类型克山病心肌间纤维化的形态学分析

目的 揭示克山病心肌间质中胶原的类型、分布、排列与含量。方法 采用本室克山病尸检材料,应用苦味酸－天狼星红染色和偏振兴显微镜观察和图像分析技术,系统地研究了克山病心肌胶原的变化。结果 在急性克山病的瘢痕灶内,胶原排列成疏网状,Ⅲ型胶原比例增加。慢性克山病的瘢痕中,胶原排列致密,１胶原比例增中Ⅰ／Ⅲ型腾的比值增大,在心肌晨坏死区域胶原一明显高于对照组,出现间质纤维化,以慢性克山病最明显。亚急性和慢性     

克山病心肌细胞线粒体钙化及其发病机理探讨

作者等最早提出肌膜和线粒体(M)的变化在克山病最先出现,本文探讨M钙化及成因。克山病心肌标本18例(急型14例,亚急型4例),包括心室壁,室中隔及乳头肌,按常规制备超薄切片,H600A型等电镜观     

p15、细胞周期蛋白D1和转移生长因子β1在非小细胞肺癌中的表达及其意义

一、材料与方法1.材料 :实验材料取自内蒙古医学院病理教研室在1996～ 1998年间的病理组织存档蜡块 ,共 16 5份。按照 1980年ＷＨＯ肺癌分类标准进行组织学分类 ,并均经病理证实。其中鳞癌 80例 ,腺癌 49例 ,不典型增生 19例 ,鳞状上皮化生17例 ;在 12 9例非小细胞肺癌中 ,同一病例手术切取标本存档蜡块不仅有肺癌组织蜡块也有淋巴结组织蜡块 ;其中淋巴结组织经ＨＥ染色证实有淋巴结转移的 35例 ,无淋巴结转移的 2 8例。其余 6 6例中 ,或同一病例手术切取标本存档蜡块仅有肺癌组织而无淋巴结组织 ,或同一病例标本存档蜡块中虽有淋巴结组织 ,…     

General primer-mediated polymerase chain reaction for detection of enteroviruses: application for diagnostic routine and persistent infections.

The aim of this study was to determine the applicability of the polymerase chain reaction (PCR) for routine diagnostic use and for the detection of persistent enteroviral infections. To this end, general primers were selected in the highly conserved part of the 5'-noncoding region of the enteroviral genome. They were tested on 66 different enterovirus serotypes. A specific fragment was amplified from 60 of 66 serotypes. An amplification product was not observed from coxsackievirus types A11, A17, and A24 and echovirus types 16, 22, and 23. Enteroviral RNA was detected by the PCR in routinely collected throat swabs and stool specimens that were found to be positive for enterovirus by isolation in tissue culture. Enteroviral RNA was detected in one of five myocardial biopsy specimens from patients with dilated cardiomyopathy, implicating virus persistence. No amplification product was obtained from eight control samples. Our results demonstrate the significance of the PCR for the detection of enteroviral RNA and, in particular, for the demonstration of persistent enteroviral infections.     

Enteroviraf RNA in endomyocardial biopsy tissues of myocarditis and dilated cardiomyopathy

Enteroviruses are potential etiologic agents of myocarditis and dilated cardiomyopathy (DCM). A recently developed molecular approach has offered evidence of viral infection by detecting the virus genome. The nested reverse transcrip-tase polymerase chain reaction (nRT-PCR) was used to detect enteroviral RNA in endomyocardial biopsy tissues of myocarditis and DCM. The authors examined 44 tissues obtained from 36 patients with myocarditis, as well as from 10 patients with non-infectious cardiac diseases as controls.
Enteroviral RNA was detected in 12 of 36 patients with myocarditis. The second endomyocardial biopsy was carried out in five of the patients, in whom enteroviral RNA was detected at the first biopsy, at intervals from 3 weeks to 8 years after the first biopsy, and enteroviral RNA was found in four and had disappeared in one. In one of the four positive patients at the second biopsy, a third biopsy was carried out 5 months later (6 months after the first), and the RNA was detected. Active myocarditis became clinically and microscopically mild at the second and third biopsies. In one patient who developed DCM, enteroviral RNA was also detected at a second biopsy performed 8 years after the first. Enteroviral infection is a probable cause of myocarditis and enterovirus-infected myocarditis may progress to DCM.     

Detection of enterovirus-specific RNA in serum: The relationship to chronic fatigue

The serum of 88 chronic fatigue patients was screened for enteroviral specific sequences by polymerase chain reaction (PCR) assay. The PCR method used was “nested” PCR targetting the 5′ nontranslated region of the enteroviral genome which yielded a final fragment length of 264 base pairs. Samples were obtained from patients during 1990–1991. In addition, buffy coat specimens and stool specimens were examined in some patients. Samples from two cohorts of comparison individuals were also obtained. The comparison groups were firstly, acutely ill individuals with symptoms consistent with a presumed enteroviral infection (matched by age, sex, and date of receipt of specimen) and secondly, healthy individuals (matched by age and date of receipt of specimen). Enteroviral specific sequences were detected in 36 of 88 serum samples from chronic fatigue patients, 22 of 82 acutely ill individuals, and 3 of 126 healthy individuals. The enteroviral PCR positivity did not correlate with any one particular feature of chronic fatigue nor did it reflect any history of illness at onset of fatigue, duration of fatigue, or age of patient. These results provide new evidence for the presence of enteroviral specific sequences in serum, buffy coat, and stool samples in many patients with chronic fatigue. This may reflect a persistent enterovirus infection in a proportion of chronic fatigue patients. © 1995 Wiley-Liss, Inc.     

Detection of enteroviral RNA by polymerase chain reaction in faecal samples from patients with aseptic meningitis.

An assay based on the polymerase chain reaction (PCR) for detection of enteroviral RNA in stool samples was carried out using specimens from 74 patients with aseptic meningitis. The primer pair and probe were derived from the highly conserved 5' non-coding enterovirus genomic region. Enteroviral RNA was detected in faeces of all 36 patients in whom an enterovirus was isolated from stool. The PCR assay yielded positive results in additionally 3/6 cases where enterovirus diagnoses were obtained by virus isolation from cerebrospinal fluid and/or serological tests. Thus, the positive outcome of the PCR assay was 39 (93%) among the 42 patients with enterovirus diagnoses. Furthermore, 7/19 (37%) cases with an etiology that was not established by other means were positive in the test indicating that the PCR assay may give considerable additional etiological information in patients with aseptic meningitis. The limit of RNA detectability in the PCR assay was about 100 TCID50 when highly cytopathogenic enterovirus types (coxsackievirus type B5 and echovirus type 11) were tested. The PCR was negative in all 13 patients with non-enterovirus diagnoses except in one case with a herpes simplex virus type 2 infection. Since enterovirus-specific IgM antibodies could be detected in this case a dual infection seemed probable. All the negative controls, included in the study, were PCR-negative and no contamination was encountered. This study proves the usefulness of the PCR assay for detection of enteroviral RNA in stool samples and suggests that the test may be an alternative to virus isolation for rapid enterovirus diagnosis in patients with aseptic meningitis.     

Detection of enteroviral infection in myocardial tissues by polymerase chain reaction (PCR)

Objective: To determine if enteroviral infection is linked to myocarditis and dilated cardiomyopathy. Enteroviruses, especially coxsackieviruses, appear to be the most common agents of viral myocarditis.
Methods: We collected 53 endomyocardial biopsies and two autopsy specimens from 41 patients affected by myocarditis or dilated cardiomyopathy. The patients were diagnosed clinically, hemodynamically, virologically and histologically (Dallas classification). We tested for the presence of enteroviral sequences by PCR, using 5' non-coding (coxsackievirus B3, CB3, map position 450–474, 584–603) derived primers. Specificity was confirmed using the Southern blot. We used a fraction of CB3 acutely infected mouse myocardial tissue as a control.
Results: We detected enteroviral sequences in four patients with active myocarditis, borderline myocarditis or cardiomyopathy. The patient with active myocarditis had shown neutralizing antibodies in serologic analysis for coxsackievirus B3 and B5.
Conclusions: The data support a weak link of enteroviral infection to human myocarditis and dilated cardiomyopathy, at least when using a PCR assay on biopsies.     

Detection of enteroviral RNA in end-stage dilated cardiomyopathy in children and adolescents

Medical records and archival myocardial specimens of 33 children and adolescents with end%stage idiopathic dilated cardiomyopathy (IDCM) were collected to evaluate retrospectively the potential role of enteroviral persistence in the pathogenesis of IDCM. The clinical history and laboratory assessment of each patient were reviewed carefully in order to obtain information on the nature and etiology of infections in the past and at the time of diagnosis of cardiomyopathy. Sixty-four formaldehyde-fixed, paraffin-embedded myocardial specimens, obtained from endomyocardial biopsies (n = 5), explanted hearts (n = 10), or autopsies (n = 49), were studied by the polymerase chain reaction (PCR) and by in situ hybridization to detect enteroviral RNA in the specimens. Control specimens included 34 formaldehyde-fixed, paraffin-embedded myocardial specimens from children with other cardiomyopathies, metabolic diseases, structural heart defects, or various noncardiac malignancies. The presence of cellular RNA in the specimens was confirmed by amplification of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA or β-actin mRNA as positive controls. Only one specimen from the 32 IDCM patients with appropriate myocardial specimens was positive for enteroviral RNA by PCR. Sequence analysis of the amplified viral segment showed a significant degree of homology between the viral sequence and echovirus 1. One specimen from the control patients also appeared positive by PCR, but sequence analysis of the amplified viral segment revealed it as rhinovirus 16. The results do not indicate any significant role for enteroviral persistence in end-stage childhood IDCM, although they need to be confirmed using a prospective study with fresh frozen specimens. However, mechanisms other than viral persistence may be more important in the progression of IDCM to end-stage heart failure in this age group. J. Med. Virol. 56:364–371, 1998 . © 1998 Wiley-Liss, Inc.     

Assessment of genotype and molecular evolution of hepatitis C virus in formalin-fixed paraffin-embedded liver tissue from patients with chronic hepatitis C virus infection

Drawbacks of hepatitis C virus (HCV) RNA detection in paraffin-embedded liver tissue have satisfactorily been solved by RT-PCR amplification of the 5'non-coding region (5'NCR). However, detection of this highly conserved region does not provide information on epidemiological or pathogenetic aspects of HCV infection. This study explores whether other functionally important genetic regions of HCV, such as the hypervariable region 1 (HVR-1) and the interferon sensitivity-determining region (ISDR), can be retrieved from paraffin-embedded liver specimens by RT-PCR, and whether the amplified material is suitable for further molecular analyses. RT-PCR amplification of 5'NCR, HVR-1, and ISDR was assessed in RNA extracted from 50 formalin-fixed, paraffin-embedded liver specimens, including 23 needle liver biopsies (11 from patients with non-A, non-B chronic hepatitis diagnosed between 1971 and 1985, 8 from subjects with normal liver histology and 4 from sequential biopsies from a patient with HCV recurrence after liver transplantation), and 27 liver explants from patients undergoing transplantation between 1988 and 1996 (16 with HCV-related cirrhosis and 11 with other disorders). The 5'NCR was successfully amplified in 8 of 11 (73%) non-A, non-B chronic hepatitis biopsies and in all of the specimens from patients with serological documentation of HCV infection. There were no false-positive results. HCV genotype was identified by RFLP analysis of the 5'NCR in the 13 cases analyzed. HVR-1 and ISDR were amplified in 24 of 28 (86%) samples, which were positive for the 5'NCR. Efficient amplification was inversely related to the time of storage. The evolutionary changes of HVR-1 and ISDR were successfully analyzed by direct sequencing of amplificates from the explanted liver and from the sequential liver biopsies in a patient with HCV infection recurrence after transplantation. These observations indicate that paraffin-embedded liver tissue, even when stored for more than 20 years, is appropriate for advanced studies on the molecular biology of HCV.