Inhibitory effects of Galidesivir(BCX4430)on rabies virus in vitro / 中国人兽共患病学报

This study analyzed the inhibitory effects of BCX4430 on rabies virus with different virulence(CVS-11 and SC16)in vitro.Direct immunofluorescence(DFA),quantitative real-time PCR and half tissue culture infective dose(TCID50)assays were used.BCX4430(250 pmol/L),compared with T-705,significantly decreased the viral titer and the relative expres-sion level of viral mRNA,similarly to the inhibitory effects of ribavirin.Thus,our study demonstrated that BCX4430 signifi-cantly inhibits the replication of rabies virus in vitro.

Research on target management of 21 medical institutions in Anhui province / 中华医院管理杂志

Objective To understand the status quo of the target management assessment of medical institutions in Anhui province, and provide a basis for further improving the performance appraisal mechanism and strategy of medical institutions. Methods In June 2017, 21 medical institutions in Anhui province conducted a questionnaire survey on the setup and implementation of target management indicators. Results Among the 21 medical institutions, there were 2, 16 and 3 hospitals in the first, second and third grades respectively. The average number of available beds was 681.35, and the average time for conducting target assessment was 4.42 years. The coverage rates of the clinical departments, medical technology departments, efficiency and efficiency, scientific research teaching, and department management of 21 medical institutions were 80% (316/397), 84% (52/63), 71% ( 120/168), 38% (48/126 ) and 60% (61/102) respectively. There were significant differences in coverage rates between clinical departments, medical technology departments, efficiency and efficiency, and research and teaching in different levels of medical institutions( P <0.05 ). Judging from the effectiveness of the current target assessment, it was considered that 64% ( 37/78 ) of the overall assessment results of the departments were in line with expectations, and the ratio of target management to hospital development management, department quality management, and functional department execution was 75% (42/64). Conclusions The status quo of performance assessment of medical institutions in Anhui province is running well, but the target management indicators and models of different medical institutions are different. It is necessary for the health administrators to establish a unified, authoritative and standard target performance assessment system.

Mechanisms of pyroptosis induced by meticillin-resistant Staphylococcus aureus in macrophages / 国际药学研究杂志

Objective: To evaluate the effect of meticillin-resistant Staphylococcus aureus(MRSA)on the pyroptosis of bone-marrow derived macrophages(BMDM)and investigate the mechanism of MRSA-induced pyroptosis in macrophages. Methods: The BMDM were triggered by the combination of lipopolysaccharide(LPS, 100 ng/ml)with adenosine-triphosphate(ATP, 3 mmol/L)or nigericin(Ng, 10 mmol/L)or treated with MRSA(multiplicity of infection 200, MOI 200)alone to induce pyroptosis in vitro. The cell morphology examination and lactate dehydrogenase(LDH)assay were applied to evaluate the cytotoxicity. The expression of pro- inflammatory cytokines, the interleukin(IL)- 1β, IL- 6 and tumor-necrosis factor(TNF)-α was detected by ELISA. The cleaved caspase-1 and mature IL-1β in the cells and released into the supernatant were detected by Western blotting. The signal pathway for the induction of pyroptosis by MRSA was investigated via the transfection of lentivirus- mediated short- hairpin RNA(shRNA)into the BMDM. Results: The treatment of BMDM with LPS/ATP, LPS/nigericin or MRSA alone caused cytotoxicity and up-regulated the expression of pro-inflammatory cytokines, IL-1β, IL-6 and TNF-α, as well as the caspase-1 activation and mature Zl-1β (P<0.01). After silencing NLRP3 or NLRC4, the expression of IL-1β induced by MRSA was significantly lessened(P< 0.01). Conclusion: MRSA could induce BMDM pyroptosis probably via activating NLRP3 inflammasome or NLRC4 inflammasome.

Modeling of H2O2-induced damage in retinal pigment epithelial cells and protective effects of blueberry anthocyanin / 中国药理学通报

Aim To establish the model of H2O2-induced damage in retinal pigment epithelial cells,and to investigate the protective effects of blueberry anthocyanin.Methods Cell viability was determined by MTT assay and the reactive oxygen species (ROS) level was measured using fluorescence assay.We selected H2O2 solution of different concentrations from 0 to 3 200 μmol · L-1 to stimulate ARPE-19 cells for 2,6 and 24 h,and compared the effects of blueberry anthocyanins of 1,5 and 10 mg · L-1 on cells pretreated with 6 h and 24 h.The model of oxidative damage was then established and applied in the next study,which focused on the protective effects of major constituents in blueberry anthocyanins,malvidin and its glycosides.Results Model condition of H2O2-induced damage in ARPE-19 cells was 800 μmol · L-1 H2O2 for 2 h.The cell viability decreased to 63.69％ when stimulated by H2O2.With pretreatment of 5 mg · L-1 blueberry anthocyanin extract (BAE),pure anthocyanin malvidin (My) and its glucoside (Mv-3-glc) and galactoside (Mv-3-gal),ARPE-19 cell viabilities were significantly improved to 86.57％,115.72％,98.15％ and 127.97％,respectively (P ＜ 0.01).Meanwhile,the ROS levels were attenuated by 52.38％,95.38％,77.55％ and 116.09％,respectively (P ＜ 0.05).Conclusion Blueberry anthocyanin could significantly inhibit the oxidative damage induced by H2O2 in retinal pigment epithelial cells,and malvidin and its glycosides contribute to the protective effects.

Epidemiology of human rabies in China, 2016 / 中华流行病学杂志

Objective: To understand the epidemiological characteristics of human rabies in China in 2016 and provide evidence for the control and prevention of human rabies. Methods: The incidence data of human rabies in China in 2016 were collected from national infectious disease reporting information management system. The surveillance data were collected from provinces of Shandong, Guizhou, Anhui, Hunan, Jiangsu and Guangxi Zhuang Autonomous Region. Excel 2013 software was used to process and summarize the data, the epidemiological characteristics of human rabies in China in 2016 were described by using indicators such as morbidity, mortality and constituent ratio. Results: A total of 644 human rabies cases were reported in 28 provinces in China in 2016, a decrease of 19.60% (157/801) compared with 2015. The provinces reporting high incidences of human rabies were Henan, Hunan, Guangxi and Guizhou, accounting for 39.44% (254/644) of the total cases. One case was reported in Qinghai province and Xinjiang Uygur Autonomous Region respectively. The male to female ratio of the cases was 2.14∶1 (439/205), and the majority of the patients were farmers (444/644). Surveillance points in 6 provinces reported 1 281 340 persons seeking post-exposure treatment, of whom 1 018 367 were treated for dog bite or scratch. A total of 764 234 persons completed the vaccination series, accounting for 63.90% (764 234/1 195 956) of the persons with grade Ⅱ and Ⅲ exposures, and 28.89% (165 677/573 571) of the persons with grade Ⅲ exposure were treated with passive immunization product. The average density of dogs in each surveillance area was 7.03/100 persons, the average canine immunization rate was 37.64%. Conclusion: The incidence of human rabies has remained decline in China in 2016, the number of the affected provinces has increased and that has the tendency of spreading to low-risk regions. The cases mainly occurred in men and farmers, and caused by dog bite or scratch. It is necessary to strengthen the health education about rabies prevention and control in rural areas and expand the coverage of canine immunization to prevent and control human rabies.

Application of three-dimensional printed skull in the teaching of human anatomy / 基础医学与临床

Objective This experiment aimed to find out the feasibility of applying 3D printing technology to anato-my education. Methods We constructed 3D printed skull using a cadaveric skull as template.26 participants were recruited from Peking Union Medical College. All participants studied cranial anatomy with 3D printed skulls as teaching materials,and then assessed practicability of 3D printed skulls by subjective assessment questionnaires. Results 57% participants considered that 3D printed skulls can present defined cranial anatomical structures.More than 90% participants agreed that 3D printed models helped spatial comprehension and study of anatomy.88% par-ticipants suggested 3D printing can solve the problem of ethics.84% participants agreed to apply 3D printed models into cranial anatomy education. Conclusions The efficacy of 3D printing had been confirmed in anatomical educa-tion. Further application of 3D printing technology in medical education needs carrying out.

Application of three-dimensional printed skull in the teaching of human anatomy / 基础医学与临床

Objective This experiment aimed to find out the feasibility of applying 3D printing technology to anato-my education. Methods We constructed 3D printed skull using a cadaveric skull as template.26 participants were recruited from Peking Union Medical College. All participants studied cranial anatomy with 3D printed skulls as teaching materials,and then assessed practicability of 3D printed skulls by subjective assessment questionnaires. Results 57% participants considered that 3D printed skulls can present defined cranial anatomical structures.More than 90% participants agreed that 3D printed models helped spatial comprehension and study of anatomy.88% par-ticipants suggested 3D printing can solve the problem of ethics.84% participants agreed to apply 3D printed models into cranial anatomy education. Conclusions The efficacy of 3D printing had been confirmed in anatomical educa-tion. Further application of 3D printing technology in medical education needs carrying out.

The Adverse Effect of the 2-1-1 Regimen for Rabies PEP in Preschool Children / 生物医学与环境科学（英文）

Post-exposure prophylaxis (PEP) has proved to be the most important measure for rabies prevention and control. There is little information regarding adverse reactions to the Essen and 2-1-1 regimens in preschool children (aged 0-6). We reexamined the outcomes of 1,109 preschool children who were vaccinated using SPEEDA under the Essen regimen between January 2011 and December 2012 and 1,267 preschool children under the 2-1-1 regimen between January 2013 and December 2014. We find that, in preschool children, the febrile reaction after the first 2-dose injection in the 2-1-1 regimen was significantly higher than that induced by the first 1-dose in the Essen procedure. Thus, we recommend that the Essen regimen should still be used for rabies PEP in preschool children.

Enhancement of Virus Replication in An Influenza A Virus NS1-Expresssing 293 Cell Line / 生物医学与环境科学（英文）

The nonstructural protein 1 (NS1) of influenza A virus, which is absent from the viral particle, but highly expressed in infected cells, strongly antagonizes the interferon (IFN)-mediated antiviral response. We engineered an NS1-expressing 293 (293-NS1) cell line with no response to IFN stimulation. Compared with the parental 293 cells, the IFN-nonresponsive 293-NS1 cells improved the growth capacity of various viruses, but the introduction of NS1 barely enhanced the propagation of Tahyna virus, a negative-strand RNA virus. In particular, fastidious enteric adenovirus that replicates poorly in 293 cells may grow more efficiently in 293-NS1 cells; thus, IFN-nonresponsive 293-NS1 cells might be of great value in diagnostic laboratories for the cultivation and isolation of human enteric adenoviruses.

Prognostic Superiority of Log Odds of Positive Lymph Nodes in Stage 3 Colorectal Cancer / 中国医学科学院学报

Objective To evaluate the prognostic value of the log odds of positive lymph nodes (LODDS) in stage 3 colorectal cancer (CRC) patients who have undergone curative resection. Methods We performed a retrospective review of 175 stage 3 CRC patients who underwent curative resection in Peking Union Medical College Hospital from 2005 to 2012. Patients were categorized respectively according to the AJCC/UICC N grade,the metastatic lymph node ratio (LNR),and the ratio of their LODDS. The relationship between the N grade,LNR,LODDS,and overall survival (OS) rates were assessed.Results The five-year disease-free survival (DFS) was significantly different among stage 3 CRC patients in different N grade (Χ(2)=33.1,P=0.000),LNR (Χ(2)=14.3,P=0.001),and LODDS (Χ(2)=14.9,P=0.001). Univariate analysis showed that TNM stage (Χ(2)=27.0,P=0.000),cancerous node(Χ(2)=3.6,P=0.040),N grade (Χ(2)=33.1,P=0.000),LNR (Χ(2)=14.3,P=0.001),and LODDS (Χ(2)=30.4,P=0.000) were related to OS. Multivariate analysis indicated that TNM stage (HR:1.84,95%CI:1.59～6.29,P=0.001) and LODDS classification (HR:1.34,95%CI:1.01～1.80,P=0.047) were independent prognostic factors for OS in stage 3 CRC patients. Conclusion LODDS is a good prognostic indicator in stage 3 CRC patients who have undergone curative resection.

Whole Genome Sequencing and Comparisons of Different Chinese Rabies Virus Lineages Including the First Complete Genome of an Arctic-like Strain in China / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To learn the rabies genome molecular characteristics and compare the difference of China rabies lineages.</p><p><b>METHODS</b>The complete genomes of 12 strains from different China rabies lineages were amplified and sequenced, and all the China street strain genomes (total 43), Arctic and Arctic-like genomes were aligned using ClustalX2, the genome homologies were analyzed using MegAlign software, and the phylogenetic trees were constructed by MEGA 5.</p><p><b>RESULTS</b>First Arctic-like rabies genome in China (CQH1202D) was reported, and we supplemented the rabies genome data of China, ensuring at least one genome was available in each China lineage. The genome size of China V (11908nt) is obviously shorter than other lineages' (11923-11925nt) for the difference of N-P non-coding regions. Among different lineages, the genome homologies are almost under 90%. CQH1202D (China IV lineage) has close relationship with strains from South Korea and they share about 95% genome similarities.</p><p><b>CONCLUSION</b>The molecular characteristics of 6 different China rabies lineages were compared and analyzed from genome level, which benefits for continued comprehensive rabies surveillance, rabies prevention and control in China.</p>

Construction and characterization of EGFP reporter gene labeled Sindbis virus / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To construct and characterize EGFP reporter gene labeled Sindbis virus (SINV).</p><p><b>METHODS</b>The reporter gene EGFP was inserted into the genome of infectious clone pBR-XJ160 by using multi-fusion long fragment PCR method. Then apply reverse genetic manipulation technique to rescue and obtain EGFP labeled SINV.</p><p><b>RESULTS</b>We successively obtained labeled SINV, which has good fluorescent expression characteristics and genetic stability.</p><p><b>CONCLUSION</b>The labeled virus can be seen in living cells and living body, and this serves as a good tool for cell and tissue tropism and biological function study of viruses. This study laid a foundation for further studying the cell tropism, biological functions and infection mechanism of SINV.</p>

Preparation and identification of a recombinant hepatitis C virus (HCV) based on sindbis virus vector / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To construct the recombinant virus-like particles containing HCV envelope glycoprotein E1E2 based on sindbis virus vector.</p><p><b>METHODS</b>The gene encoding HCV envelope glycoprotein E1E2 was cloned into sindbis virus vector to construct recombinant plasmids pBR-XJE1E2 and pVA-XJE1E2, and transfect them into BHK-21 cells to obtain recombinant virus-like particles. The expression of E1 and E2 protein were verified by Western Blot and indirect immunofluorescent assay (IFA).</p><p><b>RESULTS</b>The results of restriction enzyme digestion, PCR and sequencing analysis showed that the recombinant plasmids were constructed successfully. And the results of RT-PCR, Western blotting and IFA detection showed that the transfect cells could package HCV-like particles of expressing structural proteins E1E2.</p><p><b>CONCLUSION</b>The recombinant expression plasmids pBR-XJE1E2 and pVA-XJE1E2 based on sindbis virus vector could package HCV-like particles in eukaryotic cell, which provides a foundation for further study of its in vivo animal immune response.</p>

Current status of canine rabies in China / 生物医学与环境科学（英文）

The number of human rabies cases acquired from dog bites constitutes a high proportion of the total rabies cases in China, although the number of human rabies cases has gradually decreased in recent years. The pivotal role of dogs in the spread of rabies indicates that controlling and preventing canine rabies could be a key step in eradicating human rabies in China. The primary aims of this review are to discuss the properties and pathogenesis of the rabies virus, the clinical signs and diagnosis of canine rabies, threshold host density and vaccination of dogs, and the prevention and control of canine rabies in China.

Molecular characteristics of the genome of G I of Japanese encephalitis virus isolated from the specimen collected from viral encephalitis case for the first time / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the molecular basis of pathogenicity of Japanese encephalitis virus (JEV) by sequencing of complete nucleotide sequence and analyze the characteristics of full-length genome of genotype I Japanese encephalitis virus strains (GZ56) which was isolated from the first cerebrospinal fluid (CSF) of Japanese encephalitis patients.</p><p><b>METHODS</b>The complete nucleotide sequence was obtained by RT-PCR and sequencing was performed directly. Bioinformatics was used to analyze the nucleic acid data, deduced amino acid sequence and phylogenetic trees.</p><p><b>RESULTS</b>The result of sequence analysis showed that the genome of GZ56 strains had 10 965 nucleotides, which coded for a 3432-amino acid polyprotein. Phyolngenetic analysis based on full-length genome showed that GZ56 strains and M-28 strains which were the first isolated from mosquitoes in Yunnan in 1977 were in the same evolutionary branch. GZ56 strains belongs to genotype I of Japanese encephalitis virus, the homology of genome ranged from 96.2% to 98.6% in nucleotide and from 98.2% to 99.7% in amino acid sequences respectively when compared with selected genotype I of JEV strains in GenBank. There were 11 amino acid divergences in E protein when compared with the JEV inactivated P3 strain but they are not the key virulence sites. However, there were 14 amino acid divergences in E protein when compared with the JEV live attenuated vaccine SA14-14-2 strain and 8 amino acid divergences were the key virulence sites.</p><p><b>CONCLUSION</b>This study indicated that the full length of genome GZ56 strains had no ignificant change. It can be hypothesized from genomic level that the currently available JEV vaccines(inactivated and live attenuated) can protect against GZ56 strains infection, meanwhile, the JEV live attenuated vaccine (SA14-14-2) formulation conferred higher levels of protection.</p>

Molecular basis of one-way serological reaction between SINV and XJ-160 virus / 病毒学报

The purpose of this study is to elucidate the molecular mechanism of one-way serological reaction between XJ-160 virus and SINV by recombinant viruses which exchanged the glycoprotein genes individually or simultaneously. Three recombinant viruses were obtained based on the whole-length infectious cDNA clone of XJ-160 virus. The infectivity and pathogenesis to BHK-21 cells and animals were studied and the gene which controlled this one-way serological reaction phenomenon was searched by MCPENT. The results showed that the E2 glycoprotein was the main factor which influenced the growth rate, plaque morphology and pathogenicity of BHK-21 cells and suckling mice. The results of MCPENT showed that the E2 glycoprotein of SINV played a major role in this one-way serological reaction phenomenon. Our study identified the SINE2 gene was the determined gene for one way serological reaction between XJ-160 virus and SINV, and this research laid the foundation for further analysis of the genomic structure and function of SINV.

Effects of the site-directed mutagenesis at nsP2-726Pro on replicon vector derived from XJ-160 virus / 病毒学报

To investigate the effects of site-directed mutagenesis at nsP2-726Pro on the characteristics of replicon vector derived from XJ-160 virus, a Sindbis virus (SINV) isolated in China. The mutant vector pBRep-726L, pBRep-726S, pBRep-726V or pBRep-726A was constructed by introducing nsP2-726Pro --> Leu, nsP2-726Pro --> Ser, nsP2-726Pro --> Val or nsP2-726Pro --> Ala into XJ-160 viral replicon vector pBRepXJ respectively. To quantitatively and qualitatively determine the site-directed mutagenesis on the replicon, the recombinant plasmids expressing Neomycinr (Neo(r)), enhanced green fluorescent protein (EGFP) or Renilla luciferase (R. luc) were constructed by cloning report genes into pBRepXJ or mutant XJ-160 vector respectively. And in vitro-synthesized RNA from expression vectors were electroporated into BHK-21 cells. Compared with the wild-type replicon, the mutation nsP2-726Pro --> Val or nsP2-726Pro --> Ala accelerated the processing of CPE on BHK-21 cells and simultaneously enhanced its self-replicating capacity. The mutant vector pBRep-726L with Leu substitution exhibited similar packaging capacity to that of pBRepXJ. In contrast, pBRep-726S exhibited a medium phenotype, including the process of CPE and the activity of R. luc expression in BHK-21 cells. The site-directed mutagenesis at nsP2-726Pro not only regulates directly XJ-160 virus vector-host cell interactions, but also plays an important role in its packaging capacity. All of these results lay a basis for researching the relation between the structure and function of alphavirus genome and developing alphavirus vector system with Chinese intellectual property.

Construction and identification of replicon vector derived from an infectious full-length cDNA clone of a Sindbis virus / 病毒学报

To construct vector system of XJ-160 virus, a Sindbis virus isolated in China, recombinant vector pBRepXJ together with its helper plasmid pBR-H were derived from XJ-160 viral infectious clone pBR-XJ160 by overlap-PCR. To quantitatively and qualitatively verify the function of the replicon system, recombinant plasmids pSinRep-EGFP, pBRepXJ-EGFP, pSinRep-R and pBRepXJ-R were constructed by cloning report genes of enhanced green fluorescent protein (EGFP) or Renilla luciferase (R. luc) into pBRepXJ or pSinRep5, a commercial Sindbis vector. And in Vitro-synthesized RNA from expression vectors were electroporated into BHK-21 cells. The results indicated that the replicon vector system was capable of self-replicating in host cell, and the expression efficiency of heterologous genes corresponded with that of the commercial Sindbis vector (pSinRep5). Our study laid the basis for developing alphavirus vector system with Chinese intellectual property.

Study of the bacterial community structure of microbiota in bacterial vaginosis using amplified ribosomal DNA restriction analysis / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the bacterial community structure of the microbiota in the vaginal fluid from patients with bacterial vaginosis.</p><p><b>METHODS</b>The composition of bacteria in the samples of vaginal fluid from 3 patients with bacterial vaginosis and 1 normal premenopausal control was investigated by amplified ribosomal DNA restriction analysis(ARDRA).</p><p><b>RESULTS</b>Lactobacillus species were the predominant bacteria in the woman without bacterial vaginosis. Bacterial vaginosis was associated with higher concentrations of a variety of bacterial groups. Women with bacterial vaginosis had greater bacterial diversity, with 31 to 37 OTUs operational taxonomic units detected per sample. The species associated with bacterial vaginosis were Leptotrichia, Prevotella sp. and Megasphaera including several species with no close cultivated relatives.</p><p><b>CONCLUSIONS</b>Women with bacterial vaginosis have complex vaginal infections with many newly recognized species. ARDRA allows rapid analysis of the diversity of microorganisms in the vagina, and is capable of identifying potentially pathogenic bacteria that can not be identified by general culture.</p>

Preparation and identification of dengue virus with deletion mutation of capsid protein / 南方医科大学学报

<p><b>OBJECTIVE</b>To generate rescued viruses with deletion mutation of capsid protein from dengue virus type 2 isolated in China (DEN2-43).</p><p><b>METHODS</b>On the basis of infectious full-length cDNA clone pD212 of DEN2-43 strain virus, the deletion mutants were constructed by fusion PCR, from which the rescued viruses with deletion mutation of capsid protein were generated by transcription in vitro and electroporation.</p><p><b>RESULT AND CONCLUSION</b>Sequence analysis demonstrated that the deletion mutations had been successfully inserted into the rescued viruses obtained. These mutant viruses may hold the key for elucidating the effects of deletion mutation of capsid protein on the biological characteristics of dengue virus.</p>