Subcutaneous immunization with a novel immunogenic candidate (urease) confers protection against Brucella abortus and Brucella melitensis infections

Brucellosis is a world prevalent endemic illness that is transmitted from domestic animals to humans. Brucella spp. exploits urease for survival in the harsh conditions of stomach during the gastrointestinal infection. In this study, we examined the immune response and the protection elicited by using recombinant Brucella urease (rUrease) vaccination in BALB/c mice. The urease gene was cloned in pET28a and the resulting recombinant protein was employed as subunit vaccine. Recombinant protein was administered subcutaneously and intraperitoneally. Dosage reduction was observed with subcutaneous (SC) vaccination when compared with intraperitoneal (IP) vaccination. rUrease induced mixed Th1–Th2 immune responses with high titers of specific IgG1 and IgG2a. In lymphocyte proliferation assay, splenocytes from IP and SC‐vaccinated mice displayed a strong recall proliferative response with high amounts of IL‐4, IL‐12 and IFN‐γ production. Vaccinated mice were challenged with virulent Brucella melitensis, B. abortus and B. suis. The SC vaccination route exhibited a higher degree of protection than IP vaccination (p value ≤ 0.05). Altogether, our results indicated that rUrease could be a useful antigen candidate for the development of subunit vaccines against brucellosis.     

应用羧化胶乳凝集试验诊断人、畜布鲁氏菌病的研究

应用以化学交联法制备的布鲁氏菌１６Ｍ抗原－羧化胶乳制剂的羧化胶乳凝集试验（ＬＡＴ）、试管凝集试验（ＳＡＴ）、虎红平板凝集试验（ＲＢＰＴ）以及酶联免疫吸附试验（ＥＬＩＳＡ）、半胱氨酸凝集试验（ＣＹＴ），对在四省布鲁氏菌病疫区收集的部分人、畜血清中布氏菌抗体进行对比检测。结果表明，ＬＡＴ较之ＲＢＰＴ具有更高的特异性；与ＳＡＴ相比，无论阳性或阴性符合率均较一致。另外，ＬＡＴ可检测血清中ＩｇＧ、ＩｇＭ两类布氏菌特异性抗体。因而本试验对人、畜布病的诊断具有特异、敏感和简便的优点，更宜在基层推广应用。     

2008-2018年云南省人间布鲁菌病流行特征及灰色GM(1,1)模型预测

目的分析云南省布鲁菌病(简称布病)流行特征,建立灰色GM(1,1)模型,预测云南省布病病情。方法收集中国疾病预防控制信息系统和云南省统计局2008-2018年云南省布病疫情数据和人口资料,分析布病流行病学特征(包括时间、地区、人群分布),并以灰色GM(1,1)模型进行建模,预测2019、2020年云南省布病发病率。结果2008-2018年云南省共报告布病病例1216例,年均发病率为0.2374/10万,呈逐年递增趋势(χ²趋势=843.34,P<0.01)。病例报告主要集中在3-9月份,占总病例数的69.41%(844/1216)。病例报告数居前5位的州市分别为红河州(289例)、曲靖市(264例)、昆明市(258例)、大理州(160例)、玉溪市(134例),占总病例数的90.87%(1105/1216)。职业以农民为主,占79.03%(961/1216)。建立灰色GM(1,1)模型预测2019、2020年云南省布病发病率分别为0.4876/10万和0.4817/10万。结论云南省布病发病较以往上升,应对重点地区、重点人群进行针对性防控,并对预测结果进行前瞻性评价,逐步完善云南省布病预测模型。     

EVALUATION OF BRAINSTEM AUDITORY POTENTIAL IN BRUCELLOSIS PATIENTS WITH AND WITHOUT NEUROLOGICAL INVOLVEMENT

This study investigated the changes in brainstem auditory evoked potentials (BAEPs) in acute phase of brucellosis. Twenty-two patients with brucellosis without neurologic involvement and seven patients with neurobrucellosis were included. BAEPs were evaluated before treatment. No patient had a symptom of hearing loss. Patients who had neurologic involvement did not show any abnormalities in their BAEPs recording. This study suggests that patients with brucellosis who are in the acute phase of the disease with or without neurological involvement may have normal BAEPs. BAEPs do not seem to be a sensitive method for central nervous system involvement of brucella patients in the acute phase of the disease.     

Genetic stability of Brucella abortus S19 and RB51 vaccine strains by multiple locus variable number tandem repeat analysis (MLVA16)

The aims of the present study were (i) to assess the in vitro genetic stability of S19 and RB51 Brucella abortus vaccines strains and (ii) to evaluate the ability of multiple-locus variable-number tandem repeat (VNTR) analysis (MLVA) as a tool to be used in the quality control of live vaccines against brucellosis. Sixty-three batches of commercial S19 (n = 53) and RB51 (n = 10) vaccines, produced between 2006 and 2009, were used in this study. S19 and RB51 vaccines were obtained from, respectively, seven and two different manufacturers. Ten in vitro serial passages were performed on reference strains and on selected batches of commercial vaccines. All batches, reference strains and strains of serial passages were typed by the MLVA16. The results demonstrated that B. abortus S19 and RB51 vaccine strains are genetically stable and very homogeneous in their respective groups. Anyway, batches of S19 from one manufacturer and batches of RB51 from another presented genotypes distincts from the reference vaccine strains. In both cases, differences were found on locus Bruce07, which had addition of one repeat unit in the case of S19 batches and the deletion of one repeat unit in the case of RB51 batches. In summary, MLVA16 proved to be a molecular tool capable of discriminating small genomic variations and should be included in in vitro official tests.     

布鲁杆菌病职业人群高危行为习惯调查及健康教育效果评价

目的 调查辽西地区布鲁杆菌病(简称布病)职业人群的高危行为习惯,评估健康教育干预效果.方法 采取分层随机整群抽样的方法,2008-2010年对辽西地区与牛羊等牲畜有密切接触的职业人群高危行为习惯进行现况调查,从中抽取凌海市职业人群作为干预组,实施为期3个月的布病高危行为习惯健康教育,其余职业人群作为对照组.在健康教育基线期和终末期问卷调查干预组和对照组布病高危行为习惯情况.结果 基线期共调查916名观察对象,其中干预组499人,对照组417人.布病高危行为习惯(接触流产牲畜时防护、病畜及时扑杀、病畜深埋处理、不贩卖加工病畜、不吃病死畜肉、不喝生奶、不吃未熟乳肉制品、对饲养牲畜进行免疫)正确率干预组分别为70.9%(354/499)、64.3%(321/499)、53.8%(268/499)、92.2%(460/499)、82.2%(410/499)、87.5%(437/499)、93.6%(467/499)、56.2%(280/499),对照组分别为68.8%(287/417)、59.2%(247/417)、54.7%(228/417)、89.2%(372/417)、85.6%(360/417)、88.5%(369/417)、94.0%(392/417)、55.9%(233/417),组间比较差异无统计学意义(P均＞0.05).终末期共调查908名观察对象,其中干预组499人,对照组409人;干预组上述8种布病高危行为习惯正确率分别为85.4%(426/499)、79.0%(394/499)、69.9%(349/499)、96.4%(481/499)、94.3%(471/499)、94.9%(474/499)、94.0%(469/499)、99.4%(496/499),对照组分别为66.8%(273/409)、57.3%(234/409)、50.1%(205/409)、85.3%(349/409)、88.8%(363/409)、89.5%(366/409)、90.1%(369/409)、57.2%(234/409),组间比较差异有统计学意义(x2值分别为43.991、49.833、37.111、35.032、9.537、9.826、4.485、253.808,P均＜0.05).干预组在基线期除不吃未熟乳肉制品外,其他7种布病高危行为习惯正确率与终末期比较,差异有统计学意义(x2值分别为30.426、26.284、27.854、7.364、36.027、17.238、9270.286,P均＜0.05).结论 健康教育干预可以显著地提高布病职业人群高危行为习惯正确率,值得进行交流和推广.

Abstract：

Objective To find out demography characteristic and risk behaviors of occupation groups and to evaluate the effect of health education. Methods A baseline survey was conducted among occupation groups in the western part of Liaoning province by stratified random cluster sampling. Health education started to intervene in occupation groups in Linghai city of Liaoning province. An investigation of brucellosis related behaviors was carried out before and after a health education among occupation groups. Other occupational groups were as the control group. The research on high-risk behaviors of occupation groups was in the following eight aspects: protection taken when contact with abortion livestock, timely slaughter and burying deeply of sick animals, not selling sick animals do not eat dead meat, nor drink raw milk, do not eat raw meat, and immunity of livestock. Results Before the intervention, a total of 916 objects, of which intervention group 499 and control group 417 were investigated. The high-risk behaviors of brucellosis were not significantly different between the two groups[70.9% (354/499), 64.3%(321/499 ), 53.8% (268/499), 92.2% (460/499), 82.2% (410/499 ), 87.5% (437/499), 93.6% (467/499), 56.2%(280/499) in intervention group, 68.8% (287/417), 59.2% (247/417 ), 54.7% (228/417), 89.2% (372/417 ), 85.6%(360/417) ,885%(369/417),94.0%(392/417),55.9%(233/417) in control group, all P ＞ 0.05]. After the intervention,a total of 908 survey objects, of which intervention group 499 and control group 409, were investigated. The above eight aspects of high-risk behaviors in the intervention group were 85.4% (426/499),79.0% (394/499),69.9%(349/499),96.4%(481/499),943%(471/499),94.9%(474/499),94.0%(469/499), 99.4%(496/499), respectively; in the control 66.8% (273/409),57.3% (234/409),50.1% (205/409),85.3% (349/409),88.8% (363/409),89.5%(366/409) ,90.1% (369/409), and 57.2% (234/409), respectively. Between the two groups, the difference was statistically significant (x2 = 43.991,49.833,37.111,35.032,9.537,9.826,4.485,253.808, all P ＜ 0.05 ). The above seven aspects of high-risk behaviors except do not eat raw dairy at the beginning of intervention were compared with those of after the intervention, the differences were statistically significant (x2 = 30.426,26.284,27.854,7.364,36.027,17.238,9270.286, all P ＜ 0.05). Conclusions Health education intervention can significantly improve the behavior change rate of occupational groups at high risk of brucellosis, and the education intervention is worthwhile to communicate and promote.     

转录组分析布鲁氏杆菌患者外周血免疫基因的表达情况

目的 通过生物信息学分析方法探讨布鲁氏杆菌病的关键失调免疫基因和分子机制。方法 从GSE69597中获取布鲁氏杆菌病患者全血转录组表达数据。通过差异分析筛选布鲁氏杆菌病患者和对照之间的差异表达基因(DEGs),并利用immport数据库调取DEGs中免疫相关的基因集(immune-DEGs)。通过Enirchr在线富集工具对immune-DEGs进行富集分析。构建immune-DEGs的蛋白质-蛋白质互作(PPI)网络,并鉴定网络中的高度互联的核心(hub)基因。利用qRT-PCR验证hub基因的表达,并绘制hub基因的ROC曲线。使用ssGSEA算法评估布鲁氏杆菌病患者中免疫细胞的评分,并通过流式细胞术检测血液样本中免疫细胞的水平。结果 共获得390个immune-DEGs,富集结果中发现了T细胞受体信号通路和Th17细胞分化等。在10个hub基因中IFNG和TNF在布鲁氏杆菌组中显著高表达。ROC曲线表明IFNG对布鲁氏杆菌病具有良好诊断意义。此外,活化型CD₄ T细胞、效应型CD₄ T细胞、效应记忆型CD₈ T细胞和2型T辅助细胞因子在布鲁氏杆菌患者中明显增多。流式细胞术检测发现与健康对照组相比,布鲁氏杆菌病患者外周血中Th2和Th17细胞比例增高,Th1和Treg细胞比例则降低。结论 本研究结果不仅提高了我们对布鲁氏杆菌感染后机体免疫反应的认识,还为诊断和治疗布鲁氏杆菌病提供了更多的方向。     

9例布鲁氏菌病并发感染性主动脉瘤患者临床诊治分析 临床指标及细胞因子水平变化特征

[摘要]?目的?分析布鲁氏菌病并发感染性主动脉瘤的临床特点,以提高临床医生对该疾病的认识及诊疗水平。方法?选取2002年1月1日—2022年6月1日北京安贞医院收治的9例布鲁氏菌病并发感染性主动脉瘤患者作为研究对象,收集其流行病学资料、临床表现、实验室检查结果、影像学资料、治疗及预后等情况。结果?32例血培养证实布鲁氏杆菌感染者中,9例并发主动脉瘤,发生率为28.13%。其中8例为男性,1例为女性,年龄为58～75岁。有明确牛、羊接触史者3例,有不洁羊奶饮用史者1例,有牧区旅游史者1例。所有患者均有发热,其中热型为典型波浪热者1例。患者主要症状为腰腹痛。误诊为腰椎结核者2例,误诊为腰椎管狭窄者3例。1例患者合并附睾炎。6例患者累及腹主动脉,1例累及升主动脉,2例累及主动脉弓。9例患者血清凝集试验均阳性。9例患者均接受利福平、多西环素和左氧氟沙星抗感染治疗,除1例失访外,其余8例总疗程均为3～6个月。7例手术治疗患者中,6例行主动脉替换或修补术,1例经6个月抗感染治疗后行支架植入手术,以上患者术后均随访5个月～7年（中位时间4年）,1例于术后5年死于脑血管疾病意外,其余6例均存活。2例未接受手术治疗患者中,1例出院后失访,1例随访3年存活至今。结论?布鲁氏菌病的主动脉受累较少见,且易危及生命,由于对该病认识不足,易导致漏诊、误诊。本病治疗的基石仍然是抗感染治疗联合开放性手术。血管内支架等介入手术治疗的作用尚未确定,须要考虑患者的病情和长期并发症的风险。     

慢性布氏菌病患者中肝脏损害发生几L率的探讨

目的 探讨慢性布氏菌病（简称布病）是否可以造成肝脏损害及肝脏损害在慢性布病中发生的几率和规律。方法 对273例慢性布病患进行临床症状、体征和实验室检查三个方面的观察。结果 发现52例患有肝脏损害，占19.05%。其肝损害与布氏菌素皮试验强度有正相关关系，而与布病血清学及病程无直接关系。结论 说明慢性布病仍可造成肝脏损害，但比急性布病发生率低，其发病机理可能与Ⅳ型变态反应对肝脏的损害作用有关。