Corynebacterium pyruviciproducens is a recently described species of Corynebacterium. There are few reports on the microbiological characteristics of the new species, and there is a lack of reports on the genomic analysis of the species. This study involved a clinical isolate from the pus of a hospital patient with sebaceous gland abscesses. The clinically isolated strain was identified as C. pyruviciproducens strain WYJY-01. In this study, referring to Koch’s postulates, we observed the pathological changes of animal models infected by intraperitoneal injection and subcutaneous injection of pure culture of the strain WYJY-01. Furthermore, the strain WYJY-01 was isolated and cultured again from animal models' subcutaneous abscess drainage fluid. Subsequently, the genomics of the strain WYJY-01 was analyzed. By comparing various gene databases, this study predicted the core secondary metabolite gene cluster of the strain WYJY-01, virulence factor genes carried by prophage, pathogenicity islands, and resistance islands. In addition, the genomes of C. pyruviciproducens strain WYJY-01, ATCC BAA-1742 T, and UMB0763 were analyzed by comparative genomics, and the differential genes of strain WYJY-01 were compared, and their functions were analyzed. The findings showed that the strain WYJY-01 had pathogenicity, supplementing the phenotype characteristics of C. pyruviciproducens. Meanwhile, this research revealed the possible molecular mechanism of the pathogenicity of the strain WYJY-01 at the gene level through whole genome sequence analysis, providing a molecular basis for further research.

BACKGROUND:Gastric cancer is heterogeneous cancer and the causes of this disease are complex. New diagnostic and therapeutic targets are urgently needed to explore. Huntingtin-associated protein 1 (HAP1) is directly related to Huntington's disease (HD). However, patients with Huntington's disease have a lower incidence of cancer. Therefore, we are committed to studying the correlation between HAP1 and gastric carcinogenesis and development.METHODS AND RESULTS:Immunohistochemical staining, western blot analysis, and RT-qPCR were conducted to explore the localization and expression of HAP1 in gastric cancer. To study the biological significance of HAP1, we overexpressed HAP1 in both MKN28 and AGS cell lines by lentivirus infection. To explore the role of HAP1 in cell proliferation, the cells counting assay, EdU incorporation assay, and colony formation assay were carried out. We performed the wound healing assay and transwell assay to study the cell migration and invasion. To further investigate whether HAP1 could regulate gastric cancer cell death during glucose deprivation, Annexin V-FITC/PI staining was performed. In our study, we elucidated that HAP1 was downregulated in gastric cancer. What's more, overexpressing HAP1 inhibited cell proliferation, cell migration and invasion, and triggered apoptosis during glucose deprivation. More importantly, the antitumor properties and mechanisms of HAP1 have been elucidated further in gastric cancer.CONCLUSIONS:Taken together, the available evidence implies that HAP1 may serve as a potential tumor suppressor, making it a significant target in preventing and treating gastric cancer. This research provides a theoretical basis for the early diagnosis, clinical targeted therapy, and prognosis evaluation of gastric cancer.

Abstract Background: Pulmonary tuberculosis (PTB) affects patients' lives and reduces their quality of life. Lactoferrin (LF) is a member of the transferrin family. We explore the expression level of LF in the peripheral blood of PTB patients and the clinical significance of LF in the diagnosis and course evaluation of PTB. Methods: Plasma LF concentration and neutrophil LF concentration were measured, and their correlation was analyzed, as well as their diagnostic validity and specificity for PTB. Results: Plasma LF concentration of active PTB patients was higher than that of stable PTB patients and patients in the control group. Neutrophil LF concentration of active PTB patients was higher than that of the control group, whereas it was reversed in stable PTB patients. However, there was no significant difference in the levels of neutrophil LF and plasma LF between primary and retreated PTB patients. There was a positive correlation between them, and the LF level was positively correlated with the TB replication load. Neutrophil LF had a higher specificity and sensitivity for diagnosing stable PTB, while plasma LF had higher specificity and sensitivity in the diagnosis of active PTB. Conclusions: Plasma LF has a higher sensitivity and specificity for the diagnosis of active PTB, and LF might be a biological indicator of PTB.

Helicobacter pylori (H. pylori) is a common pathogen that infects more than half of the world's population. Its infection can not only lead to a variety of gastrointestinal diseases, such as chronic gastritis and gastric cancer (GC) but also be associated with many extra-gastrointestinal diseases. Exosomes, as a new intercellular information transmission medium, can carry biological signal molecules such as microRNAs (miRNAs) to regulate a variety of cellular physiological activities and are involved in multiple cancer processes. In this article, we provide a systematic review on the role of exosomal miRNAs in H. pylori-associated GC.

目的 采用不同的方法描述Cardiobacterium valvarum(C.valvarum)临床分离株的生物学特性,利用16S rRNA基因测序技术进行分子鉴定.方法 转种阳性血培养标本到血琼脂平板上进行细菌培育,革兰染色涂片镜检,用VITEK 2 Compact全自动微生物鉴定分析仪对临床分离株进行细菌鉴定,基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)对分离株的蛋白质进行高通量测定,E-test法对分离株作药敏试验.提取分离株的DNA,采用16S rRNA基因测序技术对PCR的产物进行测序,在NCBI的BLAST网站上与GenBank数据库上的序列做相似性比较,用MEGA7.0.26软件构建该分离株的系统进化树.结果 经细菌培养发现小而圆、光滑、不透明,灰色的菌落;经革兰染色后镜下见到小、两端圆形、革兰阴性的多形性杆菌;VITEK 2 Compact上机、MALDI-TOF-MS技术均未得到该分离株的鉴定结果;16S rRNA基因测序技术测得该菌株基因全序列约为1 450 bp,与C.valvarum F0432的16S rRNA同源性为99.59％,鉴定为C.valvarum.结论 该菌的形态、生化反应均无代表性,采用16S rRNA基因测序技术可以对C.valvarum进行鉴定,对该疾病的诊断具有重要意义.

Huntington disease (HD) is a single-gene autosomal dominant inherited neurodegenerative disease caused by a polyglutamine expansion of the protein huntingtin (HTT). Huntingtin-associated protein 1 (HAP1) is the first protein identified as an interacting partner of huntingtin, which is directly associated with HD. HAP1 is mainly expressed in the nervous system and is also found in the endocrine system and digestive system, and then involves in the occurrence of the related endocrine diseases, digestive system diseases, and cancer. Understanding the function of HAP1 could help elucidate the pathogenesis that HTT plays in the disease process. Therefore, this article attempts to summarize the latest research progress of the role of HAP1 and its application for diseases in recent years, aiming to clarify the functions of HAP1 and its interacting proteins, and provide new research ideas and new therapeutic targets for the treatment of cancer and related diseases.

目的 研究ε-聚赖氨酸对铜绿假单胞菌(Pseudomonas aeruginosa,PA)的生长及其对生物膜形成与清除的影响.方法 以P.aeruginosa PAO1、ATCC27853及收集的65株临床分离株作为受试菌,采用微量肉汤稀释法测定ε-聚赖氨酸(ε-Polylysine,ε-PL)对PA的MIC、MBC值;吸光度法测定ε-PL对PA浮游菌生长影响,并绘制时间-生长变化曲线;结晶紫染色法筛选产生物膜的临床菌株;结晶紫染色法测定ε-PL对PA产生物膜影响及对预形成生物膜的作用效果.结果 ε-PL对P.aeruginosa PAO1菌株、ATCC27853和临床分纯菌株的MIC值均为0.25 mg/ml,MBC值均为0.5 mg/ml.临床株PA生物膜阳性率为100％;时间-生长曲线表明自6 h起ε-PL对PA的抑制作用明显;随着ε-PL浓度的增加,对PA的生长、生物膜的形成抑制作用及对预形成生物膜的清除作用均逐渐增强.结论 ε-PL可抑制PA的生长和生物膜的形成,并能清除部分预形成生物膜.

目的 采用多种生物预测软件分析铜绿假单胞菌YfiB蛋白及其编码基因结构和生物学功能.方法 由NCBI数据库获取YfiB蛋白的基因组信息,经ORF Finger软件预测基因ORF区;ProtParam、SOSUI、ExPASy-ProtScale、SignalP 4.1 Server和TargetP-2.0 Server软件分析YfiB蛋白理化性质、亲疏水性和信号肽;TMHMM Server v.2.0、NetPhos 3.1 Server、BLAST软件对跨膜区域、磷酸化位点和保守结构域分析;采用SOPMA、COILS、SWISS-MODEL、ABCpred、SYFPEITHI和STRING预测软件分析YfiB蛋白结构、抗原表位及相互作用蛋白.结果 YfiB基因序列长507 bp,存在3个开放阅读框架,编码168个氨基酸.YfiB蛋白分子式为C802H1311N241O245S5,相对分子质量为18.41×103,脂族指数92.38,PI为7.89,为不稳定亲水性蛋白.含有1个信号肽,14个磷酸化修饰位点,无跨膜区域.二级结构中占比较高的是无规则卷曲和α-螺旋,分别为42.26％和33.93％.预测YfiB蛋白含有8个B细胞表位、12个限制性CTL表位和6个限制性Th表位,10种蛋白可能与其相互作用.结论 生物信息学方法预测YfiB蛋白存在多个B、T细胞表位,可为铜绿假单胞菌YfiBNR信号系统促进相关生物膜形成的功能机制研究提供参考.

碳青霉烯耐药的肺炎克雷伯菌可引起持续性感染,延长患者住院时间,增加医疗费用并造成创面经久难愈.碳青霉烯耐药的肺炎克雷伯菌不断检出给临床治疗带来极大挑战.肺炎克雷伯菌的碳青霉烯酶包括A类酶、B类酶和D类酶,不同种类的碳青霉烯治疗方案不同,故碳青霉烯酶快速及准确地检测对临床治疗具有重要意义,常用检测方法有表型检测方法和分子技术两类,综述碳青霉烯酶实验室检测方法,为碳青霉烯酶检测提供依据.

中药作为抗真菌药物,具有价格低廉、资源丰富、不良反应小、不易耐药等优点.该文通过对单味中药、复方中药、中西药协同抗真菌作用及中药抗真菌作用的机制及近年来中药抗真菌的研究现状进行综述,为临床抗真菌治疗提供理论依据.

Objective To investigate the methods for identifying Actinomyces europaeus and to analyze its biological characteristics in order to provide a basis for the diagnosis of actinomycosis. Methods Pus speci-mens collected from patients were used for bacterial culture and then analyzed with Gram staining. VITEK 2 Compact automatic microbiological analyzer was used for species identification. Drug susceptibility test was per-formed with E-test. Matrix-assisted laser desorption/ionization-time of flight mass spectrometry ( MALDI-TOF MS) was used to identify the isolated strain. The common primers of 16S rRNA were used for amplification fol-lowing DNA extraction, and the product of PCR was sequenced after recovery and purification. Homology analy-sis was conducted using the sequence in GenBank database. Results The drug susceptibility test showed that the strain was sensitive to penicillin, piperazolin/taclobatan, and ceftriaxone, but resistant to ciprofloxacin. MALDI-TOF MS and 16S rRNA gene assay identified the strain as Actinomyces europaeus. Conclusions MALDI-TOFMS and 16S rRNA could be used to identify Actinomyces europaeus and are of great significance for the diagnosis of actinomycosis.

肺炎克雷伯菌是一种革兰阴性杆菌,近年来已成为引起社区及医院感染的重要病原菌.由于抗菌药物的广泛使用甚至滥用,耐药形势日趋严峻.肺炎克雷伯菌的耐药机制主要有产生抗菌药物灭活酶、抗菌药物作用靶位的改变、抗菌药物渗透障碍、主动外排泵系统的过度表达等.本文主要讲述了肺炎克雷伯菌耐药机制的研究进展.

生物膜产生是导致铜绿假单胞菌医院难治性感染的重要原因,由于生物膜内铜绿假单胞菌难以根除,给临床治疗造成极大困扰.本文综述了菌体表面成分、胞外多糖、胞外DNA和群体感应系统参与生物膜相关铜绿假单胞菌的致病机制.简析了铜绿假单胞菌生物膜耐药性机制,介绍了通过抑制群体感应系统、抗菌肽和噬菌体等手段消除生物膜的的相关热门研究进展,为铜绿假单胞菌生物膜科研及临床相关研究提供参考.

Objective To identify a clinical isolate of Mycobacterium abscessus.Methods A pus sample was collected from a patient with suspected nontuberculous mycobacterial infection who visited the Affiliated Hospital of Weifang Medical University on December 18,2017,and was subjected to bacterial culture,Gram staining and acid-fast staining.Drug sensitivity test was conducted by the proportion method.The genome DNA of the strain was extracted and amplified by PCR with the universal primer of 16S rRNA.The PCR products were sequenced after collection and purification,and were compared with the known sequence of Mycobacterium abscessus in GenBank database.The isolate was identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS).Results The clinical isolate was identified as Mycobacterium abscessus both by MALDI-TOF MS and 16S rRNA gene sequencing.The drug sensitivity test showed that the strain was sensitive to amikacin,moxifloxacin,levofloxacin,but was resistant to streptomycin,isoniazid,rifampicin,ethambutol,ofloxacin,kanamycin,capreomycin,aminosalicylic acid,protionamide and rifabutin.The patient was diagnosed with subcutaneous soft tissue infection in the left knee joint.According to the results of drug sensitivity test,the patient was treated with amikacin and levofloxacin,and her condition was improved after treatment.Conclusion The 16S rRNA gene detection and MALDI-TOF MS both can be applied in the identification of Mycobacterium abscessus.

Objective To construct cecropin A-thanatin combinant gene engineering antimicrobial peptide gene CA(1-7)-T(4-19) for expression in Pichia pastoris.Methods The combinant antimicrobial peptide gene was artificially synthesized via gene splicing by overlap extension (SOE).The gene was cloned into the pPICZαA vector and transformed into Pichia pastoris X-33 by electroporation.The positive clones obtained by the screening of bleomycin resistance were induced by methanol ,and the antibacterial activity of the products was detected and the antimicrobial spectrum was established.Results The combinant peptide gene CA (1-7)-T (4-19) was successfully cloned on the carrier pPICZαA.The identification results were consistent with the pre-designed gene sequence.The combinant peptide gene was expressed under the induction of methanol ,and the minimum inhibitory concentration of 76 strains of Gram-egative and Gram-positive pathogenic bacteria isolated from the clinic was obtained ,and the minimum inhibitory concentration was up to 5 μg/mL.Conclusion A combinant genetic engineering antimicrobial peptide with antibacterial activity was obtained successfully and it had obvious inhibition effect on clinical common multidrug-resistant strains.

目的 探讨血清同型半胱氨酸(Hcy)、视黄醇结合蛋白(RBP)水平与冠心病及冠脉病变程度的关系.方法 选取2016年1月-2016年12月本院收治的冠状动脉造影确诊为冠心病的患者血液标本300例,以冠脉病变支数和Gensini积分评价冠脉病变严重程度,另选择健康体检者血液标本80例为对照组.分析对照组与不同病变程度冠心病患者的血清Hcy和RBP水平与冠心病之间的关系.结果 冠心病组血清Hcy、RBP水平明显高于对照组,差异有统计学意义(P＜0.05);血清Hcy、RBP水平与冠脉病变程度呈正相关(r=0.975,P＜0.05).结论 测定血清Hcy、RBP水平可以判断冠心病及冠脉病变程度.

The phenotype and genotype of the strains isolated from the sebaceous cysts were identified,and the biological characteristics and pathogenicity of the strains were described,which provided accurate etiological evidence for clinical diagnosis.Isolated strains were Gram stained and identified with VITEK 2 compact automatic microorganism analyzer.Antimiocrobial susceptibility testing was performed by K-B method.DNA was extracted from the isolated strains.The 16S rRNA fragment was amplified by universal primers PCR and sequenced.BLAST was used to compare the homology of the sequences to the 16S rRNA gene sequence in GenBank.Results showed that the isolated strain was identified as Kocuria roseus by VITEK 2 Compact,and then identified by 16S rRNA sequencing method as Corynebacteriium pyruviciproducens.Drug sensitive testing indicated that the isolates was sensitive to tetracycline,vancomycin,rifampin but resistant to penicillin,erythromycin,clindamycin,ciprofloxacin,levofloxacin,and compound new Ming,cefepime,imine pei south,gentamicin resistance.It is concluded that the phenotype of bacteria is not easy or inaccurate to identify;the 16S rRNA sequence is the most accurate determination method of identification.The Corynebacterium pyruviciproducens is a pathogenic bacterium causing the disease of the patient.At the same time,to improve the biochemical reaction information of Corynebacterium pyruviciproducens,the effective and feasible method of biological identification is also explored.

Objectives To express and purify the recombinant protein OMP25 of Brucella and to prepare a polyclonal antibody in order to lay the foundations for establishing a method of diagnosing brucellosis.Methods The OMP25 outer membrane protein of the M16 strain of Brucella was amplified using PCR.The fragment was ligated into the vector plasmid pET-30a (+) and transformed into DH5α cells.The recombinant plasmid PET-30a (+-)-OMP 25 was constructed and transformed into the Rosetta (DE3) strain.The expressed protein was collected and purified using a Ni2+NTA column purification kit.Polyclonal antibodies against OMP25 were prepared by immunizing rabbits with the recombinant protein.Results The recombinant plasmid PET-30a (+)-OMP25 was successfully constructed and transformed into the Rosetta (DE3) strain.The recombinant protein was obtained and purified.Rabbits were then immunized with the protein to prepare polyclonal antibodies against OMP25,and the polyclonal antibodies reacted specifically to the recombinant protein.Conclusions The recombinant plasmid PET-30a (+)-OMP25 was successfully constructed.Recombinant protein was expressed in Rosetta (DE3) and immunized animals produced polyclonal antibodies against the protein,indicating that the recombinant protein had good immunogenicity.Highly pure polyclonal antibodies were prepared at a high titer.This work has laid the foundation for establishing a method of diagnosing brucellosis.

目的 对与唐菖蒲伯克霍尔德氏菌和椰毒伯克霍尔德氏菌有关的4株重要的菌株进行表型性状描述,以证明它们是同一个菌种.方法 着重用传统的底物利用试验和GC及专用软件(Sherlock系统6.1)分析菌细胞的脂肪酸,做全面的表型描述.结果 4株菌的30项生化反应完全一致,170项底物利用试验95％一致,12种脂肪酸及脂肪酸组合基本一致;4株菌都是固氮菌.结论 4个株菌为同一个菌种,都可称为唐菖蒲伯克霍尔德氏菌.

为了解医学检验专业本科实习生的现状、心理特点和他们对就业的需求情况,对医学检验专业本科实习生的相关情况和现状进行问卷调查,并对获得的调查数据进行统计分析.通过调查,可掌握医学检验专业本科实习生的实习情况、对就业和考研的看法和心理健康状况,从而提出学校应进一步完善实习点和就业指导平台的建设,提供更好的就业和考研服务.