Research on international cooperation of academic papers of Peking University First Hospital / 中华医学科研管理杂志

Objective:Based on the academic papers published by Peking University First Hospital in the past 11 years, this paper analyzes the international cooperation in scientific research and aims to provide reference for the formulation of future scientific research programs.Methods:Bibliometric analysis was used to analyze the current status and trend of international cooperation, the journal distribution and preferences, as well as the subject distribution and participation.Results:It is found that the international cooperation has made some progress in recent years, however, the international cooperation rate is basically the same, while the level of cooperation should be improved. The level of international cooperation varies among different disciplines, the more strong disciplines are higher than others. Most research are concentrated in clinical medicine, basic research and other interdisciplinary research need to be strengthened.Conclusions:For the next step, we should improve the top-level design of international cooperation, strengthen the international cooperation in basic research and interdisciplinary research, and promote the development of discipline construction.

Comparative analysis of scientific research achievements between the Department of Nephrology of Peking University First Hospital and international counterparts: Based on the data of Web of Science from 2012 to 2021 / 中华医学科研管理杂志

Objective:To conduct a comparative analysis of the academic publications in the Department of Nephrology of Peking University first hospital and its international peers, to provide scientific information support for further improvement of the discipline construction.Methods:Based on Web of Science 2012-2021 paper data, international benchmarking institutions were selected for comparative analysis in terms of paper quantity, quality and impact, funding resources, etc.Results:Compared with the benchmarking institutions, the number of papers in the Department of Nephrology of Peking University first hospital increased the fastest, but the average citation frequency, the percentage of international collaborative papers, the percentage of Q1 partition papers, and other indicators ranked relatively low. The research quality needs to be further improved, while most of the research funding sources were scientific research funds.Conclusions:The scientific research level of the Department of Nephrology of Peking University First Hospital is constantly improving. In the follow-up discipline construction, we should further strengthen international cooperation, focus on high-level research content, promote industry-university-research cooperation, and carry out nephrology research with Chinese characteristics.

The development trend and hotspots analysis of international genetics research of the coronavirus from 2003 to 2021 / 中华医学科研管理杂志

Objective:To analyze the overall layout of coronavirus genetics research in the past 20 years from the perspective of publication time, journals, keywords, citations and funds, identify core research institutions and their cooperation networks in this field, and explore current research hotspots.Methods:PubMed and Web of Science databases were searched separately for the research literature and citations data about coronavirus genetics from 2003 to 2021, and Excel was used to analyze the distribution of the literature and institutions, and then the Citespace software was selected for institutional cooperation network and reference co-citation cluster analysis.Results:The literature about coronavirus genetics had increased significantly in 2020. The top 10 journals have collected 23.4% of relevant literature. Chinese Academy of Sciences has published the most documents in this field, in the top 10 high-yielding institutions, 7 are from China. In the cooperation network, the Chinese Academy of Sciences, University of Hong Kong and University of Sao Paulo have high betweenness centrality. The total litation times and average citation times of funded papers were higher than those of non-funded papers. There have been 7 research hotspots in the past 20 years. The research on " the gene sequence and functional receptors of the Middle East respiratory syndrome coronavirus" , " the gene traceability and drug development of the SARS-Cov-2" as well as " genotyping, origin and economic impact of paroxysmal porcine epidemic diarrhea virus in the United States" are still active in the past five years.Conclusions:Coronavirus genetics research will be at a sustained high level in the next few years or even longer. It is difficult to publish papers in journals with high impact factors (IF>5.00) in related fields. Chinese research institutions are active in this field. The Chinese Academy of Sciences and the University of Hong Kong are the most influential, and they have the closest cooperation with other institutions. The genetic tracing, gene sequence and functional receptor of coronavirus and drug development are likely to be the forefront of the research in this field.our govermment should advance the layout in this field and increase the number of research funds and financial support.

Research hotspots in international bioethics from 2009 to 2018: a bibliometric analysis / 中华医学科研管理杂志

Objective:To analyze the research hotspots of international bioethics, explore the future trend.Methods:The biclustering analysis was adopted to analyze the literatures of bioethics in PubMed from 2009 to 2018.Results:The research fronts from 2009 to 2018 focused on 8 aspects which including patient advocacy in clinical ethics consultation, empirical research methodologies in bioethics research, definition of death combined with the problem of organ donation.Conclusions:Greater emphasis should be placed on the combination of empirical science and the autonomy-based ethical theories for domestic bioethics studies. Furthermore, the above 8 aspects of international trends on bioethics offer a sketch for a glimpse of reflection on domestic study on bioethics.

Construction of real-time polymerase chain reaction detection for infection-related cytokines of tree shrew / 生物医学工程学杂志

Tree shrew is a novel and high-quality experimental animal model. In this study, the real-time polymerase chain reaction methods were established to detect infection-related cytokines interleukin-6 (IL-6), IL-8, IL-10, IL-17A, interferon-γ (IFN-γ) and housekeeping gene glyceraldehyde-phosphate dehydrogenase ( ) of tree shrew. The results indicated that the establised methods had good specificity. The high point of the linear range of these reagents reached 1 × 10 copies, and the low points ranged from 10 copies (IL-6, IL-17A), 100 copies (IL-10, ) to 1 000 copies (IL-8, IFN-γ). In this interval, the linear correlation coefficient of each reagent was greater than 0.99. The lowest detectable values of IL-6, IL-8, IL-10, IL-17A, IFN-γ and were 8, 8, 4, 8, 128 and 4 copies, respectively. The results showed that the established detection methods had good specificity, sensitivity and wide linear range. The methods were suitable for detection of multiple concentration range samples, and could be used for the subsequent studies of tree shrew cytokines.

Construction of human adenovirus type 4 vector expressing enhanced green fluorescence protein / 中华微生物学和免疫学杂志

Objective To prepare human adenovirus type 4 (Ad4) vector expressing enhanced green fluorescence protein (EGFP). Methods This study used a previously prepared plasmid pBRAd4 containing the whole genome DNA of Ad4-GZ01 strain. The Ad4 genome E3 region of pBRAd4 was deleted and replaced with the EGFP expression frame by conventional molecular cloning method. Then the recombi-nant plasmid was transfected into AD293 cells to rescue recombinant virus which was identified by sequen-cing,SDS-PAGE and ELISA. The purified virions were injected to mice and the induced immune responses were detected by ELISA and microneutralization test. Results The recombinant Ad4 vector rAd4EGFP ex-pressing EGFP was obtained and could be recognized and neutralized by monoclonal antibody MN4b and an-tisera against Ad4. The Ad4-specific and EGFP-specific antibodies with high titers could be detected in mice immunized with rAd4EGFP. Conclusion Human Ad4 vector expressing EGFP was successfully obtained and could be used in research on vaccine development,drug evaluation and transgene vector.

Development of international appraisal tools for clinical guidelines / 中华医学图书情报杂志

The background,features,dimension and contents of 22 international appraisal tools for clinical guidelines were analyzed,and their history,evolution,rules and problems were described,in order to provide the evidence for developing appraisal tools for domestic clinical guidelines. In the study,it was found that the guideline evaluation tools have been classified and there is a constant strive to achieve the balance between comprehensiveness and practicability. But the existing evaluation tools are insufficient for clinical practical contents of guidelines. The scoring system should be improved. In addition,conflicts of interest,value choice and patients' involvement should be considered increasingly, and should become an important part of the guideline evaluation tools.

Construction and characterization of a hexon-chimeric human adenovirus type 3 vector expressing one major epitope of dengue virus type 1 / 中华微生物学和免疫学杂志

Objective To construct a recombinant human adenovirus type 3 ( HAd3 ) vector ex-pressing one major epitope of dengue virus type 1.Methods The gene encoding the envelope protein (304-314 aa) of dengue virus type 1 was inserted into the hypervariable region 1 ( HVR1 ) of HAd3 hexon by using overlap PCR.The recombinant gene was cloned into the shuttle plasmid, then linearized with AsisⅠrestriction enzyme and co-transformed into Escherichia coli BJ5183 strains with the digested backbone plas-mid for homologous recombination.The recombinant plasmid pBRAdΔE3GFP-DENV1 was transfected into AD293 cells to rescue recombinant adenovirus strains (rAdΔE3GFP-DENV1).ELISA and Western blot as-say were performed to evaluate the humoral responses induced in BALB/c mice after the immunization with rAdΔE3GFP-DENV1 strains.Results The recombinant adenovirus strains were successfully rescued. ELISA and Western blot assay showed that the antibodies in serum sample could recognize dengue virus type 1 strains.Conclusion The recombinant adenovirus strains expressing the epitope of dengue virus type 1 were successfully constructed.This study provided evidence for the development of multivalent vaccines against dengue virus.

Construction of a hexon-chimeric human adenovirus type 3 vector expressing two neutralizing epitopes of hepatitis B surface antigen preS1 and analysis of antigenicity of the chimeric epitopes / 中华微生物学和免疫学杂志

Objective To construct a hexon-chimeric human adenovirus type 3 ( HAd3 ) vector expressing two neutralizing epitopes of hepatitis B surface antigen preS 1 (HBsAg-preS1) and to analyze the antigenicity of the chimeric epitopes .Methods Two neutralizing epitopes of HBsAg-preS1 including KR359 and KR127 were inserted into hypervariable region 1 ( HVR1) and hypervariable region 2 ( HVR2) of HAd3 hexon .Chimeric hexon gene encoding the two epitopes was amplified by overlap PCR and then subcloned in -to shuttle plasmid pBR322-L/R containing the homologous recombination regions .The digested shuttle plas-mid containing chimeric hexon gene was co-transfected into Escherichia coli BJ5183 cells together with back-bone plasmid pBRAdΔE3GFP to construct pBRAdΔE3GFP-preS1 vector.Then pBRAdΔE3GFP-preS1 vector was digested with AsiSⅠand transfected into AD293 cells to construct recombinant virus (rAD3E-preS1). CsCl gradient centrifugation was used for purification .Glutathione S-transferase ( GST ) fusion protein KR359KR127 ( GST-KR359KR127 ) was expressed in Escherichia coli BL21 by using expression vector pGEX-4T3.Female BALB/c mice at age 6-8 weeks were intraperitoneally injected with 1010 virus particles or 80 μg GST fusion protein .Serum samples were collected to analyze the antigenicity of two epitopes by ELISA and Western blot .Results ELISA showed that KR 359 and KR127 were successfully exposed on viral sur-faces by using hexon-chimeric HAd3 vector .The induced polyclonal antibodies in serum samples could rec-ognize GST fusion protein and native HBsAg from patients infected with hepatitis B virus .Conclusion The antigen capsid-incorporation strategy could be used to display epitopes on viral surface .Enhanced antigen-specific responses could be achieved through inserting multiple foreign epitopes into hexon HVRs .This study provided evidence for further application of hexon -chimeric human adenovirus type 3 vector in the developmentof vaccine, especially for the development of multivalent hepatitis B vaccine .

Cloning and expression of hexon protein of human respiratory adenovirus of three serotypes and analysis of antigenicity of the recombinant proteins / 中华微生物学和免疫学杂志

Objective To clone and express the hexon protein of three prevalent human adenovi -rus strains causing respiratory disease and analyze the antigenic characteristics of the recombinant proteins . Methods The full length genes encoding hexon protein of human adenovirus serotype 3(HAdV3), serotype 4(HAdV4) and serotype 7(HAdV7) were cloned by PCR and sequenced , respectively.The alignment anal-ysis was performed by using hexon gene sequences from GenBank .The major antigenic regions of hexon pro-tein of the three serotypes were expressed in E.coli and purified.The antigenicity, immunogenicity and cross reactivity of the recombinant proteins were determined by ELISA and Western blot assay .Results The full length gene sequence encoding hexon protein of human adenovirus serotype 4 was firstly reported in China , which showed more than 99%homology in both nucleotide and amino acid sequences with the human adeno-virus type 4 NHRC3 strain.The partial hexon protein sequence of HAdV 3, HAdV4 and HAdV7 containing all of the 7 hyperviriable regions ( HVRs) were expressed in E.coli, respectively .The purified recombinant proteins could be recognized by antiserum of the three serotypes of adenovirus .The antiserum samples against the three recombinant proteins could cross-react with particles of the three serotypes of adenovirus . The possible type-and species-specific epitopes were predicted .Conclusion The major antigenic regions of hexon protein of the three serotypes were successfully expressed .The purified recombinant proteins contai-ning both intertypes and type-specific epitopes showed a strong immunogenicity .

Investigation on evaluation of pituitary function after transsphenoidal ectomy of hypophysoma / 中华内分泌代谢杂志

Objective To assess the pituitary function in patients with pituitary adenoma after transsphenoidal ectomy of hypophysoma.Methods Data of 106 patients with pituitary adenoma who were admitted in endocrine department and underwent the operation in PLA General Hospital from January 1993 to January 2010 were collected.Assessments of pituitary function were made before and after surgery.Results Total 23.6％ and 16.0％ of 106 patients underwent pituitary function evaluation by 1 week and 3 months after surgery,respectively.23.5％ and 5.9％ of patients with hyopituitarism before surgery underwent pituitary function evaluation by 1 week and 3 months after surgery respectively,and the respective figures in those without hypopituitarism were 23.6％ and 20.8％.The incidences of new onset of hypopituitarism among 106 patients that underwent surgical procedure were 48.0％ within 1 week after surgery and 35.3％ by 3 months after surgery.Conclusion The rate of re-evaluation of pituitary function by 1 week and 3 months after pituitary surgery was low.Probably,many patients were missed the diagnosis of hypopituitarism after pituitary surgery.