Phenomenon: Laboratory safety training is very important because laboratory is the place for experimental teaching and scientific research in medical university. Due to the huge numbers of undergraduate students each year and the limited class hours, current laboratory safety training methods were not suitable to promote among them, moreover, the interdisciplinary knowledge for graduate students may beyond their understanding. Approach: We developed a laboratory safety training system specificly for undergraduate students based on Information, Motivation, and Behavioral Skills (IMB) to overcome the actual difficulties. This training system was combined with online and offline studies not only offering knowledge of laboratory safety but fulfilling the pathogenic biology experimental course. At end of the semester, Knowledge-Attitude-Practice(KAP) surveys and statistical analysis was performed. Findings: This training system had been conduced to grade 2020 and grade 2021 students of Guangzhou Medical University and received high admiration. We found that the practice(P) was positively correlated with knowledge(K) and attitude(A) (p<0.05). Aside from micro video and virtual simulation program, scientific comics were the most welcomed multimedia to assist study. Insights: Knowledges and techniques students learned from the laboratory safety training system was not only critical for their current study but will benefit their future career.

2021年4月15日《中华人民共和国生物安全法》实施,对学生进行全面、规范的实验室安全教育刻不容缓.本学院整合、优化了病原生物学实验内容并针对其中涉及的生物安全方面内容建立教育培训体系:包括线上法规学习、线上实验理论讲解、线下实际操作、线上+线下总结考试.在不增加学生课程学时的情况下培养其建立起牢固的安全意识、养成优秀的实验安全习惯.

Experimental teaching is an important part of medical undergraduate and postgraduate education. In the process of teaching and scientific research experiments, various dangerous chemicals are inevitably used. Based on the requirements of hazardous chemicals management and the current situation of hazardous chemicals in school of basic medical sciences, this paper puts forward corresponding management suggestions and countermeasures. Through the formulation of a three-level management system for university, colleges and laboratories, promoting the use of intelligent hazardous chemical cabinets, strengthening the organization of hazardous chemical emergency drills and improving the safety awareness of teachers and students, etc, the safety risks of hazardous chemicals are reduced and safety accidents do not occur during the storage and use of various hazardous chemicals.

在健康中国战略背景下,医学科普对提高居民健康管理能力及健康水平具有重要作用.MG动画凭借"短、平、快"的特点以及制作成本低的优势,已迅速成为网络科普信息传播的重要助推力,能助力医学工作者进行医学科普知识的传播,完成各类适用于各大网络平台传播的医学科普动画.但医学工作者在制作医学科普动画时,应注重根据短视频传播特点进行内容创作,兼顾医学科普内容的专业性和通俗性,以公众需求为中心进行创作,依托专业背景增强科普公信力,建设新媒体科普专业人才队伍.

传统的病原生物学实验课程是理论课的从属,实验安排往往以生物学特点作为依据,各自验证一个知识点,彼此间缺少联系;而且教授方式以老师作为主体向学生灌输相关知识.随着社会发展,知识获取的途径变得广阔而简便,实验课的教学模式也应随之改变.本文将以学生为主导,培养其能力及探索和创新精神为导向的教学模式为目标,浅析病原生物学实验课的特点以及针对这样的特点进行实验课改革的思考和方法.

Clonorchis sinensis was a food-borne zoonotic parasite in the worldwide and also an important risk factor of hepatic fibrosis. Excretory/secretion products of C. sinensis (CsESPs) are involved in parasite-host interactions and contribute to the development of hepatic damage. The aim of the present study was to investigate whether CsESPs and CsTP (adult protein) could induce autophagy of hepatic stellate cells (HSCs) and further activate HSCs so as to participate in the pathogenesis of hepatic fibrosis. The human hepatic stellate cell line LX-2 was stimulated by CsESPs and CsTP. CsESPs showed the effect on cell proliferation in methyl thiazolyl tetrazolium (MTT) assay while CsTP failed. Autophagosomes and autolysosomes were observed after the transmission mRFP-EGFP-LC3 plasmid into the LX-2 cells. CsESPs had more powerful to induce the accumulation of autophagosomes and autolysosomes to enhance autophagic flux compared with CsTP. Western-blotting analysis confirmed that the ratio of LC3-II/I in LX-2 cells was up-regulated after CsESPs treatment for 6 h, which further proved that CsESPs could induce autophagy in LX-2 cells. Meanwhile, q-PCR results showed that the mRNA levels of collagen I, collagen III and α-SMA decreased in LX-2 cells after treatment with autophagy inhibitor chloroquine, whereas they increased when combination with CsESPs. These results suggested that CsESPs-induced autophagy might be involved in the activation of HSCs, and consequently participate in the pathogenesis of hepatic fibrosis caused by C. sinensis infection.

随着社会的发展,人们对健康越来越重视,然而通过网络获取的医学科普信息常常对错参半,给广大民众造成极大的误导.所以如何帮助大家获取正确的科普知识,是高等医学院校应该承担的社会责任[1～2].高等医学院校开展科普教育活动,具有得天独厚的优势和资源[3].广州医科大学以医学标本馆为支点,辅以病原生物学基础验证实验,在组织大家观摩学习上,辅以解说人体生理功能及疾病发生发展机理,使民众能够进行正确有效的自我防护,正确应对突发急病或者公共卫生事件.同时,充分挖掘医学标本库及病原教学实验资源,把一些深奥的、专业的医学知识赋予有趣亲民的面孔,贴近百姓生活,以百姓看得懂、学得会、用得着的方式普及民众,让民众乐意积极参与科普活动.

为保证实验教学质量,实验教学技术与手段必须持续深化改革.因此,淘汰"水课",打造"金课",是提高实验教学的必然趋势.将现实场景和虚拟信息相结合呈现的增强现实技术可以把书本知识更灵活地呈现在学生面前,并且还可以通过互动功能为学生提供更好的虚拟实验条件.教师应尝试如何将增强现实技术运用到病原生物学实验课中,为打造"金课"提供更有力的支持.

Cultivating the innovative ability of university students is a main goal of practical teaching. The traditional teaching model of pathogenic biology and immunology can no longer suffice the professional educational requirements in modernized education. In order to maximize the effectiveness of experimental teaching in cultivating innovative talents, this article sets out to explore viable ways calculating college students’ innovation on the basis of pathogenic biology and immunology laboratories by constructing sharing platforms for lab teaching resources in corresponding subjects, conducting virtual lab simulations, and establishing innovation university lab, which push forward reform in laboratory teaching and construction of experiment platform.

Objective To establish a new course"Experimentation of Pathogenic Biology"for constructing systematic knowledge of pathogen including medical microbe and parasite. Methods "Experimentation of Pathogenic Biology"was implemented in 2014 and 2015 grade students major in clinical medicine in Guangzhou Medical University. Questionnaire survey was used for analyzing the efficient of this course. Results It was generally affirmed by students that this course could reflect systematic knowledge of pathogen,improve scientific research quality,interests,and operate ability,etc. Conclusion The course could integrate Medical Microbiology and Human Parasitology effectively ,and explore experiments in scope and depth.

日本血吸虫病是由血吸虫引起的一种慢性寄生虫病.本研究着力于探究日本血吸虫(Schistosome japonicum,S.j.)感染的C57BL/6小鼠肠系膜淋巴结不同固有免疫细胞的表型变化.C57BL/6小鼠感染日本血吸虫6周,分离肠系膜淋巴结,制备单细胞悬液,检测固有免疫细胞及其表面相关分子的表达情况.结果显示肠系膜淋巴结中γδT细胞、NK细胞和NKT细胞数量明显增多(P＜0.05),但只有NK细胞的百分比含量明显增加(P＜0.05);γδT细胞、NK细胞和NKT细胞表面CD69表达显著增高(P＜0.01),而CD25的变化均不明显(P＞0.05);γδT细胞和NK细胞表面CD4增高(P＜0.05);NK细胞表面NKG2A/C/E(CD94)、NKG2D (CD314)表达及NKT细胞表面NKG2D (CD314)表达降低(P＜0.05).表明日本血吸虫感染C57BL/6小鼠肠系膜淋巴结不同固有免疫细胞的表型变化存在显著差异.

The herpes simplex virus 1 (HSV-1) UL31 protein is a multifunctional nucleoprotein that is important for viral infection; however, little is known concerning its subcellular localization signal. Here, by transfection with a series of HSV-1 UL31 deletion mutants fused to enhanced yellow fluorescent protein (EYFP), a bipartite nuclear localization signal (NLS) was identified and mapped to amino acids (aa) 1 to 27 (MYDTDPHRRGSRPGPYHGKERRRSRSS). Additionally, fluorescence results showed that the predicted nuclear export signal (NES) might be nonfunctional, and the functional NES of UL31 might require a specific conformation. Taken together, these results would provide significant information for the study of the biological function of UL31 during HSV-1 infection.

Codon usage bias among synonymous codons is not an uncommon phenomenon and it is known to involve various biological factors, such as GC compositions, gene length, mutation frequency and patterns, gene expression level, etc. Knowledge on synonymous codon usage may help in understanding the molecular evolution of the individual gene better. In the present study, we examined the codon usage bias between pseudorabies virus (PRV) EP0 gene and the EP0-like genes of 24 reference alpha herpesviruses. Comparative analysis showed noticeable disparities of the synonymous codon usage bias in the 25 alpha herpesviruses, indicated by codon adaptation index, an effective number of codons (ENc) and GC3s value. The codon usage pattern of PRV EP0 gene was phylogenetically conserved and similar to that of the EP0-like genes of the genus Mardivirus of alphaherpesvirus, with a strong bias towards the codons with C and G at the third codon position. Cluster analysis of codon usage pattern of PRV EP0 gene with its reference alpha herpesviruses demonstrated that the codon usage bias of EP0-like genes of 25 alpha herpesviruses had close relation with their gene functions. ENc-plot revealed that the genetic heterogeneity in PRV EP0 gene and the 24 reference alpha herpesviruses was constrained by G+C content, but not gene length. In addition, comparison of codon preferences in the EP0 gene of PRV with those of E. coli, yeast and human revealed that there were 40 codons showing distinct usage differences between PRV and yeast, 27 between PRV and E. coli, but only 22 between PRV and human. Therefore, the human expression system may be more suitable for expression of PRV EP0 gene. In conclusion, these results may improve our understanding of the evolution, pathogenesis and functional studies of PRV.

在本科临床专业留学生医学微生物学教学过程中，根据课程大纲的具体要求、各章节的内容以及学生的学习能力，联合运用PBL和LBL教学法，教学效果显著提高，获得学生广泛认可，为留学生教育改革的全面深入开展提供了思路。

目的 确定肺炎链球菌自溶酶(LytA)蛋白B细胞线性表位.方法 通过生物信息学方法预测肺炎链球菌LytA蛋白分子的B细胞表位并人工合成相应肽段;经克隆、表达、纯化等步骤得到纯化的重组LytA(rLytA)蛋白用以免疫小鼠并进行Westen blot鉴定;取免疫组小鼠和对照组小鼠血清分别与人工合成的候选B细胞表位肽段进行间接ELISA检测,筛选出B细胞表位.结果 利用多种方法分析LytA蛋白分子的B细胞表位,经整理得到11条候选肽段,命名为LB1到LB11,并进行了人工合成.利用分子生物学技术获得了rLytA蛋白并成功地免疫了小鼠,间接ELISA结果显示:肽段LB3、LB5、LB6、LB7、LB11与rLytA免疫小鼠的血清发生反应,其OD 450 nm读数比相应的阴性对照小鼠显著升高,差异有统计学意义(P＜0.01).结论 确定了肺炎链球菌LytA蛋白分子5个B细胞线性表位的位置和序列.

Objective To establish acute kidney disease mouse model caused by aristolochic acid Ⅰ.Methods The mice were divided into experimental groups and control group,experimental groups were respectively given one-offinjection of aristolochic acid Ⅰ 20,50,100 mg/kg body weight in mice,which leaded to acute renal injury in mice.Took blood and kidney tissue samples on the 3rd,7th day of the experiment,tested related renal function indexes and tissue morphology.Results After the injection of aristolochic acid Ⅰ,the contents of creatinine,blood urea nitrogen of mice increased,the kidney slice staining showed kidney tissue morphological changes were associated with dose and time.Conclusion Application of large dose of aristolochic acid Ⅰ successfully established a mouse model of acute kidney disease.

[目的]确定肺炎链球菌表面粘附素A(PsaA)蛋白抗原的B细胞、CD4 T细胞和CD8 T细胞表位.[方法]通过生物信息学方法预测肺炎链球菌PsaA蛋白分子小鼠B细胞线性表位、MHC-Ⅰ结合肽(CD8 T细胞表位)及MHC-Ⅱ结合肽(CD4 T细胞表位)并人工合成相应肽段;克隆重组质粒BL21/pET/PsaA并得到纯化的rPsaA蛋白用以免疫小鼠;分离每只免疫组和对照组小鼠的血清分别与人工合成的候选B细胞表位肽段进行间接ELISA检测,筛选B细胞表位;分离小鼠脾及淋巴结细胞并分别与人工合成的T细胞候选表位肽段体外共培养,取上清进行双夹心ELISA检测IFN-γ的量进行初步筛选.用初筛到的表位多肽刺激小鼠的脾及淋巴结细胞并进行细胞内染色及细胞流式检测,确定PsaA蛋白分子T细胞表位.[结果]分析PsaA蛋白分子的B细胞表位及MHC-Ⅰ结合肽、MHC-Ⅱ结合肽;得到候选B细胞表位肽10条,MHC-Ⅰ结合肽21条,MHC-Ⅱ结合肽7条,并进行了人工合成.利用分子生物学技术获得了rPsaA蛋白并成功免疫了小鼠.间接ELISA结果显示:肽段PB2、PB6,PB7与rPsaA免疫小鼠的血清发生反应,其OD450nm读数比相应的阴性对照小鼠显著升高;双夹心ELISA结果初步提示:肽段PⅠ 10、PⅠ 14、PⅠ 17、PⅠ 18、PⅠ21以及肽段PⅡ2、PⅡ5、PⅡ6刺激免疫小鼠细胞产生的IFN-γ量比相应的对照小鼠显著升高.细胞流式结果进一步确定:经肽段PⅡ2、PⅡ5刺激后,免疫小鼠细胞中IFN-γ和CD4双阳性细胞百分比较对照小鼠细胞显著升高;经肽段PⅠ 14、PⅠ 17、PⅠ21刺激后,免疫小鼠细胞中IFN-γ和CD8双阳性细胞的百分比较对照小鼠细胞显著升高.[结论]确定了肺炎链球菌PsaA蛋白分子的3个B细胞线性表位;2个CD4 T细胞表位;3个CD8 T细胞表位.

目的 研究伯氏疟原虫CSP基因的N端蛋白与小鼠肝细胞膜HPSGs受体结合后在体外、体内的肝组织靶向性.方法 将伯氏疟原虫CSP基因N端片段(PbCSP-N)克隆入原核表达质粒pET-28a,经IPTG诱导表达重组蛋白,经亲和层析纯化后采用Western blot方法鉴定;采用Pull-down方法检测PbCSP-N蛋白与小鼠肝细胞表面受体硫酸乙酰肝素糖蛋白(Heparan sulfate proteoglycan,HSPG)的结合力;应用免疫组化、放射性碘标记示踪方法检测该蛋白在体外、体内特异性靶向小鼠肝组织作用.结果 成功构建PbCSP-N原核表达质粒并表达、纯化出PbCSP-N重组蛋白(15 ku),Pull-down方法证实该蛋白可与小鼠肝细胞膜HPSGs受体结合,免疫组化试验显示该重组蛋白定位于正常小鼠肝细胞膜,表明该蛋白可特异结合正常小鼠肝细胞膜;放射性碘(125I)标记示踪显示,标记的重组蛋白在注射1、2、4、6、12 h后在小鼠肝脏分布较多,表明该重组蛋白可靶向正常小鼠肝组织.结论 伯氏疟原虫CSP基因N端蛋白与肝细胞膜HPSGs受体结合后在体内、外靶向肝细胞,可作为肝细胞靶向性药物候选分子.

In order to evaluate immunogenicity and protective efficacy of LytA from Streptococcus pneumoniae, we subcloned the full-length lytA-encoded autolysin (LytA) from 5 major pathogenic serotype isolates in China and obtained purified rLytA. Bioinformatics analysis showed that sequences of LytA were highly conserved in all strains we used in this work, and western blot analysis demonstrated that rLytAs from heterogeneous serotypes were cross-recognized by serum of mice infected with 23F strain SH137. Mice were intranasally immunized with purified rLytA, and serum anti-rLytA IgG, IgA and secretory IgA were elicited. More importantly, rLytA intranasal-immunized mice showed a significantly higher survival rate and lower bacterial carriage in response to infection by Streptococcus pneumoniae. The fact that mice immunized with rLytA from strain SH137 also had a higher survival rate after intraperitoneal injection of other four serotype strains of living S. pneumoniae suggested that it possessed cross-protection effect. Our study revealed that intranasal immunization with rLytA may protect mice against mucosal and systemic pneumococcal infection; hence, it was an attractive vaccine candidate.