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关键词： 牛A群轮状病毒   单链抗体   VP7蛋白   单克隆抗体   原核表达   活性鉴定  

摘要： 旨在制备靶向牛A群轮状病毒(Group A Bovine Rotavirus, BRVA)VP7蛋白的单链抗体(single-chain fragment variable, scFv).以项目组前期实验制备的BRVA VP7蛋白鼠源单克隆抗体C8杂交瘤细胞为模板,通过高通量测序获得其抗体可变区(variable region, V区)基因序列,并使用(Gly_(4)Ser)^(×)3作为linker构建scFv基因片段,并将其克隆至pET28a(+)载体,转化至大肠杆菌BL21(DE3)中表达.结果显示,C8-scFv在大肠杆菌BL21(DE3)中以包涵体形式表达,其相对分子质量约为26 kDa.免疫荧光试验结果表明,C8-scFv能够特异性识别BRVA,具有良好的反应活性.研究成功制备了靶向BRVA VP7蛋白的scFv,为开发基于免疫学的BRVA检测技术奠定了基础.

关键词： X连锁疾病   XK基因   慢性肉芽肿   Mcleod综合征   棘型红细胞   抗-KL  

摘要： 目的探讨罕见Mcleod表型伴X连锁慢性肉芽肿的血液学上的特点,KEL和XK基因分析,患儿不规则抗体检出的鉴定思路和高频抗原抗体的血清学特性以及输血指导策略。方法利用血清学方法对患儿ABO、Rh及其它血型系统抗原表型的鉴定及不规则抗体的筛查和鉴定,高频抗原抗体的效价采用试管抗人球法和微柱凝胶卡式法测定,Kell血型采用血清学和基因分型技术,XK基因测序采用二代测序技术;患儿母亲外周血涂片镜检红细胞形态。结果患儿血清学结果:血型O型、ccDEE、MM型、Le(a-b+)、JK(a+b+)、Fy(a+b-),Kell血型:K-k+、Kp(a-b+),血浆中检出同种抗体抗-C、e和高频抗原抗体抗-KL,抗-KL的效价为:512(试管法),2048(微柱凝胶法);基因型结果:KEL基因分型:K-k+、Kp(a-b+)、Js(a-b+),XK基因高通量测序结果为XK∗N.01,exon1~3半合子缺失,XK:-1或Kx-(Mcleod);患儿母亲外周血涂片可见明显的棘状红细胞。结论罕见Mcleod表型伴X连锁慢性肉芽肿患儿的血液学特征为Kell抗原的弱表达,XK基因为全外显子缺失表型,建议临床应重点关注儿童早期X染色体连锁的慢性肉芽肿患儿的临床症状及实验室诊断,尽早为此类患儿进行XK基因鉴定McLeod表型,谨慎输血治疗,避免无法找到相合血液制品,危及患儿生命。

关键词： T淋巴细胞   白癜风   血小板活化因子(PAF)   疾病活动  

摘要： 目的探讨白癜风患者血清血小板活化因子(PAF)水平、T细胞免疫功能与疾病活动的关系。方法选择2022-07-18/2023-07-26期间在我院就诊的白癜风患者102例作为研究对象,根据白癜风疾病活动度评分(VIDA)将患者分为进展期(n=54)和稳定期(n=49),对比两组患者PAF及T淋巴细胞水平,Logistic回归分析PAF水平与疾病活动的关系及其与T细胞亚群的相关性。采用非条件Logistic回归模型分析PAF水平与T细胞亚群在疾病活动中的交互作用。结果进展期和稳定期组患者CD3^(+)水平无显著差异,进展期组患者PAF及CD8^(+)水平显著高于稳定期组患者,CD4^(+)、CD4^(+)/CD8^(+)水平则显著低于稳定期组;PAF水平评估白癜风患者疾病活动,PAF水平为18.24 ng/L时约登指数(0.670)最高,对应灵敏度和特异度分别为84.22%、82.77%,受试者工作特征(ROC)曲线下面积AUC为0.858;PAF水平与疾病活动的关联强度呈非线性剂量-反应关系(非线性检测);当VIDA评分≥1时,不同PAF水平白癜风患者CD4^(+)及CD8^(+)水平具有显著差异,不同VIDA评分白癜风患者不同PAF水平间CD4^(+)/CD8^(+)均具有显著差异;交互作用分析调整混杂因素后PAF水平与T细胞亚群对白癜风患者疾病活动影响在相加模型[相对超额危险度(RERI)=4.674,95%置信区间(CI):1.032~11.942;交互作用归因比(AP)=0.763,95%CI:0.336~1.201;交互作用指数(S)=6.854,95%CI:1.904~16.520)]和相乘模型[比值比(OR)相乘=3.461,95%CI:1.365~8.713)]中存在交互作用。结论白癜风患者血清PAF、CD4^(+)、CD8^(+)水平及CD4^(+)/CD8^(+)比值与疾病活动密切相关,且PAF水平与T细胞亚群(CD4^(+)、CD8^(+)、CD4^(+)/CD8^(+))在白癜风患者疾病活动中存在交互作用。PAF水平对白癜风患者疾病活动具有一定评估价值。

关键词： 支气管哮喘   肺炎支原体感染   肿瘤坏死因子-α   内皮素-1  

摘要： 目的 探讨支气管哮喘并肺炎支原体感染患儿肿瘤坏死因子-α(NF-α)、内皮素-1(ET-1)水平在其疾病进展评估中的意义。方法 选择支气管哮喘并肺炎支原体感染患儿100例作为观察组,并根据疾病严重程度将患儿分为轻度组(40例)、中度组(33例)以及重度组(27例);另选择健康体检儿童100例作为对照组。检测并比较对照组与观察组TNF-α以及ET-1水平;比较观察组不同病情严重程度(轻度组、中度组、重度组)患儿TNF-α、ET-1水平;分析TNF-α以及ET-1水平与支气管哮喘并肺炎支原体感染疾病进展的相关性。结果 观察组TNF-α(103.80±30.24)ng/L以及ET-1(79.58±18.75)ng/L均明显高于对照组的(0.46±0.11)、(0.72±0.14)ng/L(P<0.05)。轻度组TNF-α、ET-1水平分别为(76.32±9.18)、(50.57±0.14)ng/L,中度组分别为(103.24±17.53)、(80.79±0.23)ng/L,重度组分别为(145.18±25.16)、(121.08±0.34)ng/L;随着病情严重程度上升,患儿TNF-α、ET-1水平均有所升高(P<0.05);重度组和中度组TNF-α、ET-1水平均高于轻度组(P<0.05);重度组TNF-α、ET-1水平均高于中度组(P<0.05)。Pearson相关分析显示:ET-1以及TNF-α水平与患儿病情严重程度呈正相关(r=0.487、0.492, P<0.05)。结论 支气管哮喘并肺炎支原体感染患儿TNF-α以及ET-1水平均有明显上升,且ET-1、TNF-α与患儿病情严重程度存在密切关系,可作为临床判断患儿病情严重程度的主要指标。

关键词： 血小板   CD36   基因   献血者   血小板资料库  

摘要： 目的建立东莞地区CD36抗原缺失型血小板献血者资料库并进行特点分析。方法对565名纳入建库标准的献血者血样本采用流式细胞术进行血小板CD36抗原检测,对13例CD36抗原缺失型和187例纳入建库标准的献血者血样本采用三代基因测序分析技术进行基因序列检测。结果流式细胞术共检出血小板CD36抗原缺失型13例,缺失型率为2.30%(13/565),其中Ⅰ型缺失型2例,占15.38%(2/13),Ⅱ型缺失型11例,占84.62%(11/13);CD36抗原缺失型率在性别、民族和ABO血型分布比较均无显著性差异(P>0.05);13例CD36抗原缺失型和187例CD36正常型血样本经三代基因序列测序,13例CD36缺失型中有7例发生基因突变,其中5例在外显子,2例在内含子区域,2例Ⅰ型未检出基因突变,发生基因突变均为Ⅱ型缺失型,且均为杂合子。187例CD36正常纳入建库标准献血者共检出27例CD36基因突变,CD36缺失型基因突变率(53.85%,7/13)高于CD36正常型基因突变率(14.44%,27/187)(P<0.05)。结论本地区血小板CD36抗原缺失型频率除高于重庆地区外,与国内其他地区报道基本一致,适合采用血清学方法在本地区建立一支CD36抗原缺失型血小板献血者资料库,保障临床输血安全。

关键词： bidirectional effect   breast cancer   cancer   heart failure   immune system   mechanisms   CVD   cardiovascular disease   HF   heart failure   IL   interleukin   TAC   transverse aortic constriction   sEV   small extracellular vesicle  

关键词： killer cell immunoglobulin-like receptors   KIR-HLA-I mismatch   natural killer cells   renal transplant   microvascular inflammation   antibody-mediated rejection   Ab   antibody   AMR   antibody-mediated rejection   CI   confidence interval   CMV   cytomegalovirus   HLA   human leukocyte antigen   HLA-I   HLA class-I   iKIR   inhibitory killer cell immunoglobulin-like receptor   iKIR-MM   iKIR-HLA-I mismatch   IQR   interquartile range   KT   kidney transplantation   KTR   kidney transplant recipients   MVI   microvascular inflammation   NK   natural killer   PBMC   peripheral blood mononuclear cell  

摘要： Inhibitory killer cell immunoglobulin-like receptors (iKIRs) are randomly expressed by natural killer (NK) cell subsets and recognize motifs shared by HLA class-I (HLA-I) allotypes. Such interactions prevent NK cell autoreactivity while enhancing their response against cells lacking those HLA-I molecules ( missing self ), a situation defined in transplantation as iKIR-HLA-I mismatch (iKIR-MM), whose genotypic prediction has been associated with microvascular inflammation (MVI). Herein, we compared iKIR-MM in kidney transplant recipients with MVI ≥2 (n = 19) and controls with MVI ≤1 (n = 36). In parallel to genetic analysis of iKIR-MM, which was more frequent in MVI ≥2 patients, putative alloreactive iKIR-MM NK cells were defined by flow cytometry as NKG2A(−) cells bearing self-specific but lacking donor-specific iKIR. Although iKIR-MM NK cells were detected in both groups, their pretransplant numbers were higher in MVI ≥2 patients (median = 11.02, interquartile range = 0-58.31 vs median = 0, interquartile range = 0-9.46), especially in the presence of donor-specific antibodies or C4d, and correlated with MVI grade. Pretransplant, a subset of MVI ≥2 patients showed high proportions and numbers of oligoclonal iKIR-MM NK cells, which displayed an NKG2C(+) adaptive phenotype associated with cytomegalovirus infection. This pilot study provides a novel perspective on the contribution of iKIR-MM NK cells to MVI, with potential practical implications.

关键词： Acute kidney injury   Drug causality   Drug reaction with eosinophilia and systemic symptoms   Human leukocyte antigens   Latency   RegiSCAR   Severe cutaneous adverse drug reactions   Vancomycin  

关键词： 胫骨横向骨搬移术   糖尿病足溃疡   免疫球蛋白   创面愈合机制   截肢率  

摘要： 目的探讨胫骨横向骨搬移术(tibial transverse transport,TTT)治疗Wagner 3~4级2型糖尿病足溃疡的临床疗效以及免疫球蛋白水平变化趋势。方法回顾分析2022年5月—2023年9月采用TTT治疗的68例(73足)Wagner 3~4级2型糖尿病足溃疡患者临床资料。其中男49例,女19例;年龄44~91岁,平均67.3岁。糖尿病足溃疡Wagner分级为3级40例、4级28例。2型糖尿病病程5~23年,平均10年。观察并记录创面愈合例数、愈合时间、截肢例数、死亡例数及并发症发生情况。收集TTT治疗糖尿病足的6个关键时间点[术前1 d及术后3 d、7 d(向上横搬移第1天)、14 d(向下横搬移第1天)、21 d(搬移结束后第1天)、36 d(搬移装置去除后第1天)]血清标本,采用流式细胞术检测血清免疫球蛋白水平,包括免疫球蛋白G(immunoglobulin G,IgG)、IgA、IgM、IgE、补体C3(complement C3,C3)、C4、免疫球蛋白轻链κ(immunoglobulin light chainκ,KAP)、免疫球蛋白轻链λ(immunoglobulin light chainλ,LAM)。结果术后68例患者均随访至6个月。术后发生钉道感染3例、切口感染2例。其中5例患者(7.4%)于术后59~103 d截肢、8例患者(11.8%)于术后49~77 d死亡;余55例患者(80.9%)创面均愈合,愈合时间48~135 d,平均80 d。随访至6个月无溃疡复发、截骨周围骨折及截骨区域皮肤坏死发生。55例创面愈合患者血清免疫球蛋白水平检测示,与术前比较,术后3、7 d IgG、IgM水平显著下降(P<0.05),14 d后逐渐恢复至术前水平,36 d达峰值。IgA水平随时间延长持续下调,各时间点与术前比较差异均有统计学意义(P<0.05)。IgE水平变化表现出显著的时间异质性,术后21 d较术前明显下降(P<0.05);其他时间点均高于术前,但差异无统计学意义(P>0.05)。C3水平呈现明确的治疗相关性升高,术后7、14、21 d较术前显著提升(P<0.05),峰值出现在14 d。C4水平变化趋势与C3基本一致但变化幅度较小,术后7、14 d与术前比较差异有统计学意义(P<0.05)。KAP/LAM水平手术前后各时间点差异均无统计学意义(P>0.05)。结论TTT可加速创面愈合,有效治疗糖尿病足溃疡,降低截肢率,临床疗效确切。初步发现TTT治疗糖尿病足溃疡的潜在机制包括,通过动态调控IgG、IgA、IgM和IgE水平平衡炎症与修复过程,C3、C4水平的阶段性升高可能促进组织清理、血管再生和抗感染防御。

关键词： Autoantibodies   Integrin αvβ6   Primary Sclerosing Cholangitis   Prognosis   Ulcerative Colitis   ALP   alkaline phosphatase   anti-αvβ6   anti-αvβ6 autoantibodies   AST   aspartate aminotransferase   AUC   area under the ROC curve   CALiD   Consortium for Autoimmune Liver Disease   CCL11   C-C motif chemokine 11   CCL20   C-C motif chemokine 20   CD   Crohn’s disease   CDCP1   CUB domain-containing protein 1   CD5   T-cell surface glycoprotein CD5   CD244   natural killer cell receptor 2B4   CI   confidence interval   CV-AUC   AUC based on 10-fold cross-validation   ELISA   enzyme-linked immunosorbent assay   GDNF   glial cell line-derived neurotrophic factor   IBD   inflammatory bowel disease   IL-8   interleukin 8   IL-10   interleukin 10   IL-17A   interleukin 17A   IQR   interquartile range   LSM   liver stiffness measurement   Mayo-ES   Mayo Endoscopic Score   MMP-10   matrix metalloproteinase-10   MRCP   magnetic resonance cholangiopancreatography   MRE   magnetic resonance elastography   MSH   Mount Sinai Hospital   NPX   normalized (relative) protein expression   PD-L1   programmed cell death ligand 1   PSC   primary sclerosing cholangitis   ROC   receiver operating characteristic   SCF   stem cell factor   SD   standard deviation   SES-CD   Simple Endoscopic Score for Crohn’s Disease   TB   total bilirubin   TGF   transforming growth factor   TNF   tumor necrosis factor   UC   ulcerative colitis   UC Davis   University of California at Davis   VCTE   vibration-controlled transient elastography  

摘要： Background & Aims Anti-integrin αvβ6 autoantibodies (anti-αvβ6) are found in more than 50% of individuals with ulcerative colitis (UC). We aimed to determine the prevalence of anti-αvβ6 in patients with primary sclerosing cholangitis (PSC) and their association with liver disease severity. Methods Four cohorts of pre-liver transplant patients with PSC were recruited. Patients with inflammatory bowel disease (IBD) and healthy controls (HCs) served as comparators. Total IgG and anti-αvβ6 levels were measured using enzyme-linked immunosorbent assay. Olink inflammation panel was run on a subset of samples. Multivariable linear regression analysis was performed to assess the association between anti-αvβ6 and indices of liver disease severity. Results A total of 137 patients with PSC (including 76 with PSC-UC, 33 with PSC-Crohn’s disease (CD), and 28 with PSC alone) and 160 controls (including 91 with IBD and 69 HCs) were enrolled. Anti-αvβ6 levels were significantly higher in PSC-UC and PSC-CD compared with PSC alone ( P < .0001 and P < .003) and HCs ( P < .0001 and P < .0001). However, anti-αvβ6 levels in PSC alone were not increased compared with HCs. In patients with PSC-IBD, anti-αvβ6 levels correlated with markers of liver disease severity, including alkaline phosphatase level (r = 0.32; P = .004), the revised Mayo PSC risk score (r = 0.25; P = .02), and liver stiffness measurement (r = 0.43; P = .008) after adjusting for age, gender, race/ethnicity, and IBD subtype. Additionally, anti-αvβ6 levels were associated with markers of systemic inflammation and tissue remodeling. Conclusion Anti-αvβ6 autoantibodies identify a subset of patients with PSC with concomitant IBD.