抗大肠癌噬菌体人单链抗体的初步鉴定及序列分析

目的 :对抗大肠癌细胞的单克隆噬菌体单链抗体进行初步鉴定和测序分析。方法 :采用细胞ELISA ,免疫组化 ,DNA序列测定和计算机分析方法 ,对 5个单克隆噬菌体抗体 (CH2 73,CH2 0 5 ,CH2 0 9,CHA12 ,CH72 3)进行初步鉴定和序列分析。结果 :5个抗体均对人大肠癌细胞、人胚肾上皮细胞和其它某些人肿瘤细胞反应 ,也与人正常肝细胞有弱阳性反应 ,但不与鼠源性的癌细胞和正常细胞反应。细胞免疫组化进一步证实了ELISA结果的正确性。大肠癌免疫组化对大肠癌组织有特异性的结合反应 ,而不与正常大肠组织反应。测序结果为CH2 73ScFv全长 732bp ;V ,D ,J分别属于VH3 30 D1 2 6 JH3 linker V1 13 JL2 ,GenBank序号为AY0 2 8777和AY0 2 8996 ;CH2 0 5全长 36 6bp ,V ,D ,J分别VH1 4 6 D6 13 JH3,GenBank序号为AF35 936 5 ;CH2 0 9,CHA12和CH72 3的ScFv基因完全相同 ,全长 72 3bp ,其VH DH JH与CH2 73ScFv基因中的VH DH JH 完全一致 ,V ,D ,J分别属于VH3 30 D1 2 6 JH3 linker L2 Jκ2 ,GenBank序号为AF36 3774。结论 :噬菌体抗体具有结合人大肠癌组织和细胞的活性 ,为进一步开发临床应用人源抗肿瘤抗体和小分子抗体片段奠定基础     

Increased occurrence of free immunoglobulin light chains in cerebrospinal fluid and serum in human immunodeficiency virus-1 infection

The presence of free immunoglobulin light chains (FLCs) in the cerebrospinal fluid (CSF) and sera of patients with human immunodeficiency virus-1 (HIV-1) infection, multiple sclerosis (MS), and neurologically healthy control individuals was investigated by paying special attention to ensure that only truly free light chains would be detected. The FLCs were extracted by specifically binding them to Sepharose-coupled anti-FLC monoclonal antibodies, and thereafter they were electrophoresed and immunoblotted with monoclonal antibodies to both light chain (LC) isotypes. A frequent occurrence of kappa and lambda FLCs was found in both CSF and sera of HIV-1 infected patients. In HIV-1 infection and in MS, the frequency of FLCs of the CSF was equal. In healthy controls, only occasional weak FLCs were observed in either CSF or serum. FLC bands of the CSF from patients with HIV-1 infection tended to be more intensive than those of the appropriately diluted sera. Both intrathecal synthesis of FLCs and their transudation from sera through the impaired blood-brain barrier (BBB) may contribute to this. Increasing severity of general HIV-1 infection was accompanied by an increase of FLC intensity in sera. A qualitative demonstration of FLC in the CSF may be meaningful only in the absence of altered BBB function.     

Differentiation of Helicobacter pylori isolates by polymerase chain reaction—restriction fragment length polymorphism

Objective: To investigate the association between the diversity of urease gene and urease activity of clinical isolates of Helicobacter pylori (H. pylori). Methods: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of urease gene and rapid urease activity test were used to study the urease activity of different clinical isolates of H. pylori. Results: H. pylori clinical isolates were divided into 4 types according to their PCR-RFLP results of urease gene and urease activity. Type Ⅰ , possessing strong urease activity (0. 11) and presented 1 fragment of 1. 7 kb by PCR-RFLP, had close relations with gastric ulcer; type Ⅱ , with the weakest urease activity (0. 07) and 2 fragments (1. 3 and 0. 4 kb respectively) , was associated with duodenal bulb ulcer; type Ⅲ , with the strongest urease activity (0. 12) and 2 fragments (0. 4 and 0. 17 kb) with or without 1 fragment (0. 23 or 0. 37 kb) , was responsible for gastritis; type Ⅳ , with weak urease activity (0. 09) and 2 fragments     

环孢菌素A和1,2 5(OH)2D3防治实验性自身免疫性甲状腺炎的研究

将猪的甲状腺球蛋白（pTG)100μg/只分别于第0d,第14d皮下注入CBA小鼠体内，制作实验性自身免疫性甲状腺炎（EAT）的动物模型。免免疫干预组从0－28d，治疗组从10－38d单独或者联合应用环孢菌素A（CsA,10mg/kg)灌胃和（或）1,25(OH)2D3(0.2μg/kg）腹腔注射，pTG免疫后第28d，第38d处死小鼠，取甲状腺组织作病理学检查，并检测血清中猪的甲状腺球蛋白抗体（pTGAb)、猪的甲状腺微粒体抗体（pTMAb)。免疫干预组和治疗组联合应用小剂量CsA和1，25（OH）2D3分别使EAT发病率降低44．44％和37．50％。严重病例分别降低71．43％和60．32％，免疫干预组的血清pTGAb,pTMAb的值均降低。提示：小剂量免疫抑制剂CsA和1，25（OH）2D2联合防治EAT有效，并具有协同作用。     

微卫星改变与肺癌发生发展的关系

目的 :研究肺癌组织中第 3号染色体短臂 (3P)和第 9号染色体短臂 (9P)上微卫星标志物的改变与肺癌发生发展的关系。方法 :应用聚合酶链反应 (PCR)结合微卫星银染法检测 5 6例肺癌和 2 3例良性肺部病变 3P和 9P上的微卫星标志物的改变。结果 :5 6例肺癌组织标本中 3P上有微卫星改变 2 6例 ,检出率 46% ,9P上有微卫星改变的 2 3例 ,检出率为 41% ,5 6例肺癌组织中 ,3P和 9P上有微卫星改变的 3 9例 ,检出率 69 6% ,2 3例肺部良性疾病组织中均未出现微卫星标志物改变。肺癌组织标本中 3P上的微卫星改变与肺癌的分化程度有关 ,低分化肺癌的微卫星改变较高、中分化肺癌的微卫星改变明显增多 (P <0 .0 1)。而 9P上的微卫星改变与淋巴结转移有关 ,有淋巴结转移的肺癌组织标本上微卫星改变较不伴淋巴结转移的微卫星改变检出率明显增多 ,(P <0 .0 5 )。结论 :3P和 9P上的微卫星改变与肺癌发生、发展和转移密切相关 ,检测肺癌组织 3P和 9P上的微卫星改变对肺癌的早期诊断和判断有无淋巴结转移有一定的价值     

甲型H1N1流感若干共同关注的问题

今年3月18日,墨西哥首次报道了人感染甲型H1N1流感病毒,很快这种疾病就从美国与墨西哥交界处往世界蔓延,全球立刻拉响了甲型H1N1流感警钟[甲型H1N1流感,原称人感染猪流感,WHO于2009年4月30日更名为A(H1N1)流感,我国相继更名为甲型H1N1流感].     

Immune modulation by cadmium and lead in the acute reporter antigen-popliteal lymph node assay.

Immune modulation by heavy metals may cause serious adverse health effects in humans, although the mechanisms involved are not well understood. Both cadmium and lead are important environmental and occupational toxins. Therefore, in the current study, the costimulatory/adjuvant effects and the T-cell-activating potential of these metals (i.e., CdCl2 and PbCl2), are examined. These immune-modulating properties are critical in the development of conditions such as allergy, hypersensitivity, and autoimmunity. Using the direct popliteal lymph node assay (PLNA) and reporter antigen-popliteal lymph node assay (RA-PLNA) both metals were examined individually for immunotoxicity. Mercury (i.e., HgCl2) was included for comparative purposes as its effects in the RA-PLNA are well documented. Seven days following a single footpad injection containing metal and/or RA (trinitrophenyl-ovalbumin [TNP-OVA] or TNP-Ficoll), BALB/c mice were sacrificed and the popliteal lymph nodes (PLNs) removed. PLN cellularity, TNP-specific antibody-secreting cells (ASCs), and lymphocyte subsets were assessed. All three metals strongly stimulated T- and B-cell proliferation and ASC production following coinjection with the RA TNP-OVA. In each case, ASC production was skewed towards the IgG1 isotype. In addition, all three metals induced IgG production to TNP-Ficoll (although relatively weakly in the case of Cd). These results show that each of these metals can provide adjuvant signals to promote lymphocyte proliferation and enhance adaptive immune responses to unrelated antigens. Skewing of immune responses towards T helper type 2 responses suggests that each of these metals can enhance allergic and hypersensitivity reactions to environmental antigens. Furthermore, the induction of IgG responses to TNP-Ficoll, a T-cell-independent antigen, indicates that each of these metals can activate neoantigen-specific T cells. T-cell activation by metals can lead to metal hypersensitivity and has been implicated in the development of autoimmunity. This is the first report of immune modulation by CdCl2 and PbCl2 in the RA-PLNA.     

Presence of beta human papillomaviruses in nonmelanoma skin cancer from organ transplant recipients and immunocompetent patients in the West of Scotland

Background  Nonmelanoma skin cancer (NMSC) has been linked to cutaneous human papillomaviruses of the genus beta (betaPV).
Objectives  We sought to assess the presence of betaPV in NMSC biopsies from a group of Scottish skin cancer patients, both immunocompetent (IC) patients and immunosuppressed (IS) organ transplant recipients.
Methods  One hundred and twenty-one paraffin-embedded skin tumours (27 actinic keratosis, 41 intraepidermal carcinoma, 53 squamous cell carcinoma) and 11 normal skin samples were analysed for the presence of betaPV by a polymerase chain reaction–reverse hybridization assay designed to detect the presence of the 25 known betaPV genotypes.
Results  In IC patients, betaPV was detected in 30 of 59 (51%) tumours and two of 11 (18%) normal skin samples ( P  =   0·046). In IS patients, betaPV was found in 27 of 62 (44%) tumours; no normal skin samples were available for comparison. The most frequently found genotypes were HPV-24, HPV-15 and HPV-38. Of those tumours infected with betaPV, 28 of 57 (49%) were infected with more than one genotype (range 2–8). Tumours from IS patients were from a younger age group (mean age 57·4 years) than IC patients (mean age 73·8 years). Multiple infections were more common in tumours from IC patients (21 of 30; 70%) compared with those from IS patients (seven of 27; 26%) ( P  <   0·001). In the IC group, age did not appear to influence the distribution of single and multiple infections whereas in IS patients the proportion of multiple infections to single infections increased with age. There were no multiple infections in normal skin.
Conclusions  A wide spectrum of betaPV types was detected in our samples. Further characterization of betaPV in vivo is needed in order to determine the mechanisms by which the virus contributes to cutaneous carcinogenesis.