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关键词： 牙龈卟啉单胞菌   FimAⅡ菌毛蛋白   FimAⅡ型特异性单克隆抗体   特性鉴定  

摘要： [目的]诱导表达、纯化牙龈卟啉单胞菌(Porphyromonas gingivalis,Pg)FimAⅡ蛋白,将其作为免疫原制备抗Pg FimAⅡ单克隆抗体(mAbs),对抗体特性进行初步研究。[方法]将BL21-pET28a-FimAⅡ菌株诱导表达,镍柱亲和层析纯化重组蛋白并作为抗原免疫BALB/c小鼠,经PEG融合法和腹水法获得FimAⅡ单克隆抗体;分别采用SDS-PAGE法、酶联免疫吸附实验检测抗体纯度、效价和亚型,Western Blot和Biacore检测抗体的特异性和亲和力。[结果]FimAⅡ重组蛋白为可溶性表达,最适诱导条件为1 mmol/L IPTG诱导12 h;纯化后的重组蛋白纯度为90%以上,该免疫原制备的FimAⅡ单克隆抗体为IgG1亚类,kappa亚型,效价为1:8×104以上,特异性良好,亲和力可达到9nM。[结论]成功诱导表达、纯化FimAⅡ蛋白并制备抗Pg FimAⅡ特异性单克隆抗体,为后续开发快速检测Pg诊断试纸条奠定基础。

关键词： HLA DP401   HLA-DRA   humanized mice   MHC II   Staphylococcus aureus pneumonia   transgene  

摘要： Background:Staphylococcus aureus can cause serious infections by secreting many superantigen exotoxins in“carrier”or“pathogenic”states.HLA DQ and HLA DR humanized mice have been used as a small animal model to study the role of two molecules during S.aureus infection.However,the contribution of HLA DP to S.aureus infection is unknown yet.Methods:In this study,we have produced HLA DP401 and HLA DRA0101 humanized mice by microinjection of C57BL/6J zygotes.Neo-floxed IAβ+/-mice were crossbred with Ella-Cre and further crossbred with HLA DP401 or HLA-DRA0101 humanized mice.After several rounds of traditional crossbreeding,we finally obtained HLA DP401-IAβ-/-and HLA DRA-IAβ-/-humanized mice,in which human DP401 or DRA0101 molecule was introduced into IAβ-/-mice deficient in endogenous murine MHC classⅡmolecules.A transnasal infection murine model of S.aureus pneumonia was induced in the humanized mice by administering 2×108CFU of S.aureus Newman dropwise into the nasal cavity.The immune responses and histopathology changes were further assessed in lungs in these infected mice.Results:We evaluated the local and systemic effects of S.aureus delivered intranasally in HLA DP401-IAβ-/-and HLA DRA-IAβ-/-transgenic mice.S.aureus Newman infection significantly increased the m RNA level of IL 12p40 in lungs in humanized mice.An increase in IFN-γand IL-6 protein was observed in HLA DRA-IAβ-/-mice.We observed a declining trend in the percentage of F4/80+macrophages in lungs in HLA DP401-IAβ-/-mice and a decreasing ratio of CD4+to CD8+T cells in lungs in IAβ-/-mice and HLA DP401-IAβ-/-mice.A decreasing ratio of Vβ3+to Vβ8+T cells was also found in the lymph node of IAβ-/-mice and HLA DP401-IAβ-/-mice.S.aureus Newman infection resulted in a weaker pathological injury in lungs in IAβ-/-genetic background mice.Conclusion:These humanized mice will be an invaluable mouse model to resolve the pathological mechanism of S.aureus pneumonia and study what role DP molecule plays in S.aureus infection.

关键词： EB病毒   传染性单核细胞增多症   腺苷脱氨酶   白细胞介素-10   Toll样受体9  

摘要： 目的分析血清腺苷脱氨酶(ADA)、白细胞介素-10(IL-10)及Toll样受体9(TLR9)与EB病毒(EBV)相关传染性单核细胞增多症(IM)患儿EBV-DNA载量的相关性。方法前瞻性选择2021年2月至2022年12月就诊于该院的98例EBV相关IM患儿作为IM组,另选同期于该院体检的50例健康儿童作为对照组。比较对照组、IM组血清ADA、IL-10及TLR9水平,线性回归分析ADA、IL-10、TLR9水平与IM发生的关系,绘制受试者工作特征(ROC)曲线分析血清ADA、IL-10、TLR9水平单独及联合预测IM发生的价值。根据EBV-DNA载量不同将IM组患儿分为低载量组(18例)、低-中载量组(25例)、中-高载量组(49例)、高载量组(6例)4个亚组,比较各组血清ADA、IL-10及TLR9水平,Spearman等级相关分析血清ADA、IL-10、TLR9水平与IM患儿EBV-DNA载量的关系。结果与对照组比较,IM组血清ADA、IL-10、TLR9水平升高(P<0.05);线性回归分析结果显示,ADA、IL-10、TLR9水平均与EBV相关IM的发生有关(P<0.05);ROC曲线结果发现血清ADA、IL-10、TLR9水平单独及联合预测EBV相关IM发生的曲线下面积分别为0.850(95%CI 0.791~0.910)、0.867(95%CI 0.811~0.924)、0.891(95%CI 0.840~0.943)、0.912(95%CI 0.843~0.956);血清ADA、IL-10、TLR9水平及EBV-DNA载量从高到低为高载量组、中-高载量组、低-中载量组、低载量组(P<0.05);Spearman等级相关结果显示,血清ADA(r=0.526,P<0.001)、IL-10(r=0.610,P<0.001)及TLR9(r=0.715,P<0.001)水平与EBV相关IM患儿EBV-DNA载量呈正相关。结论EBV相关IM患儿血清ADA、IL-10及TLR9水平异常升高,上述指标水平与EBV-DNA载量密切相关,且三者联合可有效预测EBV相关IM的发生。

关键词： antibody   cancer therapy   drug delivery   nanoparticle   TNFR  

摘要： Tumor necrosis factor receptors(TNFRs)are promising targets for cancer therapy.However,activating their downstream signaling requires cross-linking of TNFRs.Herein,to devise strong agonists of TNFRs,ligands targeting TNFRs,such as OX40L and tumor necrosis factor-related apoptosis-inducing ligand(TRAIL),were fused with a multivalent protein scaffold(MV)to prepare multivalent agonists for cross-linking TNFRs.The nano-clustered multivalent-OX40L(MV-OX40L)and MV-TRAIL could promote T cell activation and directly induce tumor cell apoptosis.Moreover,to develop a universal nano-adaptor for the rapid preparation of multivalent agonists of different TNFRs,the Fc receptor that could immobilize antibodies was fused with MV to prepare MV-FcR,which could multimerize commercial agonist antibodies targeting TNFRs,such as anti-OX40 antibody(αOX40).Simply incubatingαOX40 with MV-FcR could prepare MV-αOX40 to enhance its antitumor efficacy.In addition,MV-FcR could multimerize with other therapeutic antibodies,such as anti-PD-L1 antibody,to enhance their valency.This study provides a promising strategy for engineering multivalent antitumor protein drugs.

关键词： IgG4相关性疾病   胸腔积液   诊断  

摘要： 免疫球蛋白G4相关性疾病(immunoglobulin-G4 related disease,IgG4-RD)是一类免疫介导的慢性炎症伴纤维化的疾病,对这一疾病的认识最初始于自身免疫性胰腺炎[1]。现已发现IgG4-RD可累及多个器官,而胸部受累尤其是孤立的胸部受累在IgG4-RD患者中并不常见[2],其中表现为胸膜病变且以胸腔积液为主要临床表现的病例更为少见。产生胸腔积液的病因众多,鉴别诊断复杂,容易误诊、漏诊。

关键词： 盆腔炎   慢性   桂枝茯苓丸   血清炎症   血液流变学   机体免疫力  

摘要： 目的:探讨桂枝茯苓丸对慢性盆腔炎(chronic pelvic inflammatory disease,CPID)患者血清炎症因子、血液流变学、机体免疫力的影响。方法:选取CPID患者139例,通过暗箱抽号法随机分为对照组和观察组,对照组66例,观察组73例。对照组给予甲硝唑口服,观察组在甲硝唑口服的基础上给予桂枝茯苓丸口服;比较两组患者临床症状改变情况,通过实验室检查两组患者血清中超敏C反应蛋白(hypersensitive C-reactive protein,hs-CRP)、白细胞介素8(Interleukin-8,IL-8)、白细胞介素10(Interleukin-10,IL-10)、转化生长因子β1(transforming growth factor-β1,TGF-β1)、巨噬细胞集落刺激因子(granulocytemacrophage colony stimulating factor,GM-CSF)、血液流变学、补体C3表达水平。结果:治疗后,观察组血清指标hs-CRP、IL-10、TGF-β1、GM-CSF、补体C3较对照组明显降低(P<0.05),观察组IL-8表达水平较对照组显著升高(P<0.05);观察组红细胞沉降率、血浆黏度、血浆黏度高切、血浆黏度低切、红细胞比积较对照组显著下降(P<0.05);观察组临床症状积分(主症、次症)较对照组显著降低(P<0.05);观察组总有效率[91.78%(67/73)]高于对照组[81.81%(54/66)](P<0.05)。结论:甲硝唑配合桂枝茯苓丸口服可有效降低患者血清指标炎症、降低血浆黏稠度、改善临床症状、增强机体免疫力。

关键词： 甲状腺结节   TBSRTCⅢ细胞   免疫组化   基因检测  

摘要： 探讨免疫组化与基因检测在甲状腺结节US-FNAC活检TBSRTC Ⅲ细胞患者良恶性鉴别的应用价值。方法 回顾性选取我院FNA甲状腺结节细胞学诊断为TBSRTC Ⅲ细胞病例并通过后期手术得到病理诊断患者133例,采用免疫组化(CK-19、Galectin-3、HMBE-1)、基因检测(BRAFV600E)等不种方式对良恶性不明的TBSRTCⅢ细胞进行检测,与术后病理对照分析不同检测指标诊断效能。结果 免疫组化诊断恶性76例,良性9例,术后病理恶性92例,良性41例,灵敏度82.6%,特异度78%、准确性81.2%;BRAFV600E基因突变83例,均为恶性,40例基因未发现突变,术后良性病例41例,灵敏度90.2%,特异度97.5%、准确性92.5%。两种方式联合诊断灵敏度可达96.1%,特异度84.7%。结论 免疫组化与基因检测均可提高TBSRTCⅢ细胞患者良恶性鉴别,通过以上方式不同条件医院得到更加精准的TBSRTCⅢ细胞良恶性鉴别诊断方法。

关键词： mesoporous silica nanoparticle   reactive oxygen species   ceria nanoparticle   nanozyme   poly(acrylicacid)   inflammatory bowel disease   INDUCED MOUSE MODELS   MECHANISMS   APOPTOSIS   DELIVERY   THERAPY   TNF   ROS  

摘要： Inflammatory bowel disease (IBD) is characterized by an inappropriate and persistent inflammatory immune response and is often accompanied by excessive reactive oxygen species (ROS) production. For effective IBD treatment, there is a high demand for safe and targeted therapy that can be orally administered. In this study, we aimed to propose the use of inflamed colon-targeted antioxidant nanotherapeutics (ICANs) for in situ oxidative stress level modulation in colitis. ICANs consist of mesoporous silica nanoparticles (MSNs) with surface-attached ROS-scavenging ceria nanoparticles (CeNPs), which are further coated with poly-(acrylic acid) (PAA) to facilitate preferential adherence to inflamed colon tissues through electrostatic interaction. We achieved a high ROS-scavenging property that remained effective even after artificial gastrointestinal fluid incubation by optimization of the molecular weight and PAA-coating pH. The orally administered ICANs demonstrated enhanced adherence to inflamed colon tissues in an acute inflammation mouse model of IBD induced by dextran sulfate sodium. This targeted delivery resulted in gut microenvironment modulation by regulating redox balance and reducing inflammatory cell infiltration, thereby suppressing the colitis-associated immune response. These findings highlight the potential of noninvasive ICANs as a promising candidate for treating inflammatory intestinal diseases by oxidative stress level modulation in colitis.

关键词： 晚期非小细胞肺癌   程序性死亡受体-1抑制剂   培美曲塞   卡铂   血清肿瘤标志物  

摘要： 目的 探讨程序性死亡受体-1(PD-1)抑制剂治疗肺癌患者的疗效及对T淋巴细胞亚群的影响。方法 选取2018年10月至2021年9月苏州市第九人民医院收治的晚期非小细胞肺癌患者110例作为研究对象,按照随机数字表法分为观察组和对照组,各55例。对照组给予培美曲塞联合卡铂化疗方案治疗,观察组加用PD-1抑制剂治疗,比较两组疗效、免疫功能指标、血清肿瘤标志物水平、不良反应以及随访1年生存情况。结果 与对照组比较,观察组治疗有效率较高,中位总生存时间、中位无进展生存时间较长,差异有统计学意义(P<0.05)。与对照组比较,观察组治疗后CD3^(+)、CD4^(+)、CD4^(+)/CD8^(+)较高,CD8^(+)较低,癌胚抗原(CEA)、糖类抗原125(CA125)及糖类抗原199(CA199)水平较低,差异有统计学意义(P<0.05)。两组患者不良反应发生率比较差异无统计学意义(P>0.05)。结论 晚期非小细胞肺癌采用PD-1抑制剂联合培美曲塞与卡铂化疗方案治疗的近期效果显著,患者的免疫功能指标及肿瘤标志物水平得到了明显的改善,治疗期间不会增加不良反应,中位生存时间及中位无进展生存时间较长,综合预后表现更好。

关键词： 慢性胃炎   益气化浊解毒汤   中医证候   T淋巴细胞亚群   不良反应  

摘要： 目的探究益气化浊解毒汤治疗慢性胃炎的疗效及对患者T淋巴细胞亚群表达的影响。方法选取2019年7月至2021年7月湖北省地质职工医院诊治的72例慢性胃炎患者作为研究对象,按照随机数字表法分为对照组与观察组,每组36例。对照组给予常规西医综合治疗,观察组在对照组基础上加以益气化浊解毒汤治疗。比较两组临床疗效、中医证候积分、T淋巴细胞亚群表达水平及不良反应发生情况。结果观察组治疗总有效率为94.44%,高于对照组的77.78%,差异有统计学意义(P<0.05)。治疗前,两组胃脘疼痛、胃脘胀满、纳差、嗳气积分比较差异无统计学意义;治疗1个月,两组胃脘疼痛、胃脘胀满、纳差、嗳气积分均低于治疗前,且观察组低于对照组,差异有统计学意义(P<0.05)。治疗前,两组CD3^(+)、CD4^(+)、CD8^(+)水平比较差异无统计学意义;治疗1个月,两组CD3^(+)、CD4^(+)水平均高于治疗前,CD8^(+)水平均低于治疗前,且观察组CD3^(+)、CD4^(+)水平均高于对照组,CD8^(+)水平低于对照组,差异有统计学意义(P<0.05)。两组不良反应发生率比较差异无统计学意义。结论益气化浊解毒汤治疗慢性胃炎效果确切,能减轻患者中医症状,改善T淋巴细胞亚群表达,提高机体免疫力,且无严重不良反应,安全性较高。