Objective To express human B7. 2 extracellular domain with prokaryote expression system and to evaluate its biological activity in vitro. Methods PCR was used to amplify the extracellular region of human B7. 2 which c...Objective To express human B7. 2 extracellular domain with prokaryote expression system and to evaluate its biological activity in vitro. Methods PCR was used to amplify the extracellular region of human B7. 2 which contained both the IgV and IgC domains. The recombinant PGEX-4T-3/hB7. 2 (IgV+C) was obtained by cloning the PCR product into a prokaryote expression plasmid PGEX-4T-3 and was transformed into the host strain of DH5-a. Tke fusion protein consisted of GST and hB7. 2(IgV+C) was identified by SDS-PAGE and Western blotting. T cell activation was observed by exposing purified T lymphocytes to the fusion protein and [3H]-TdR incorporation with the presence of the first signal imitated hy anti-CD3 antibody. Results The fusion protein GST-hB7. 2 (IgV+ C) was produced and detected in inclusive body form from engineered bacterial cells. With the first signal existed,T lymphocytes proliferated when it was co-stimulated by the fusion protein. Conclusion These results indicated that the functional human B7. 2(IgV+C) fusion protein can be produced in bacterial cells and the fusion protein displays the co-stimulatory activity in T lymphocytes activation.展开更多
Since 2021,clade 2.3.4.4b H5N1 highly pathogenic avian influenza viruses(AIVs)have become the dominant strain responsible for the ongoing panzootics in birds worldwide[1],causing unprecedented deathsofpoultry and wild...Since 2021,clade 2.3.4.4b H5N1 highly pathogenic avian influenza viruses(AIVs)have become the dominant strain responsible for the ongoing panzootics in birds worldwide[1],causing unprecedented deathsofpoultry and wild birds(https://wahis.-woah.org).Moreover,the H5N1 viruses have caused increasing infections in domestic and wild mammals[2].展开更多
BACKGROUND Prevalence of the main rheumatic diseases in the Republic of Sakha(Yakutia)[RS(Y)],one of the regions of the Russian Federation,differs from the other regions of the Russian Federation due to its ethnic and...BACKGROUND Prevalence of the main rheumatic diseases in the Republic of Sakha(Yakutia)[RS(Y)],one of the regions of the Russian Federation,differs from the other regions of the Russian Federation due to its ethnic and geographic features.Knowledge regarding the prevalence and structure of juvenile idiopathic arthritis(JIA)allows us to shape the work of the pediatric rheumatology service in the region correctly,and optimize the healthcare system and the need for medica-tions.AIM To describe the epidemiological,demographic,clinical,and laboratory characteristics of children with JIA in the RS(Y)and evaluate the main outcomes.METHODS This retrospective cohort study assessed all the data from the medical histories of the patients(n=225)diagnosed with JIA(2016-2023)in the Cardiorheumatology Department of the M.E.Nikolaev National Center of Medicine.Pearson'sχ²test,Fisher's exact test,Mann–Whitney and Kruskal-Wallis tests were used for statistical analyses.RESULTS The ethnic prevalence of JIA is higher in Sakha than in Russian children at 110.1 per 100000 children and 69.4 per 100000 children,respectively.The prevalence of JIA among boys and girls in Sakha was similar,unlike in Russians,where the number of girls predominated.The JIA categories were as follows:(1)Systemic arthritis:3.5%;(2)Oligoarthritis(persistent and extended):33.8%;(3)Rheumatoid factor(RF)(+)polyarthritis:0.9%;(4)RF(-)polyarthritis:14.7%;(5)Enthesitis-related arthritis(ERA):44%;and(6)Psoriatic arthritis:3.1%.Prevalence of the ERA category was 4.4 times higher in Sakha children,but the prevalence of systemic arthritis was 2.9 times lower compared to Russians(P=0.0005).The frequency of uveitis was 10.2%,and the frequency of human leukocyte antigen(HLA)B27 was 39.6%in JIA children.Biologic treatment was received by 40.4%of JIA children and 45.3%achieved remission.CONCLUSION Higher JIA prevalence,male and ERA predominance,related to a higher frequency of HLA B27 are typical in RS(Y).These data might improve the pediatric rheumatology health service.展开更多
Primordial germ cells(PGCs),the precursors of oocytes and spermatozoa,are highly pluripotent.In recent years,the in vitro induction of human primordial germ cell-like cells(h PGCLCs)has advanced significantly.However,...Primordial germ cells(PGCs),the precursors of oocytes and spermatozoa,are highly pluripotent.In recent years,the in vitro induction of human primordial germ cell-like cells(h PGCLCs)has advanced significantly.However,the stability and efficacy of obtaining h PGCLCs in vitro still require improvement.In the current study,we identified a novel induction system using Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12(DMEM/F-12)as the basal medium,supplemented with B27 and N2(referred to as N2B27)in combination with four cytokines:bone morphogenetic protein 4,stem cell factor,epidermal growth factor,and leukemia inhibitory factor.The h PGCLCs induced under these conditions closely resembled PGCs from 4-to 5-week-old embryos at the transcriptomic level.Compared with traditional GK15(GMEM supplemented with 15%Knockout?Serum Replacement)-based induction conditions,the N2B27 system significantly increased the speed and efficacy of h PGCLC induction.RNA sequencing analysis revealed that this improvement was due to an increased cellular capacity to cope with hypoxic stress and avoid apoptosis.The N2B27 medium promoted enhanced mitochondrial activity,enabling cells to better manage hypoxic stress while reducing the production of reactive oxygen species.Moreover,through gradient concentration experiments,we demonstrated that the addition of the common antioxidant N-acetyl-L-cysteine at an optimized concentration further enhanced the efficiency of PGCLC induction under GK15 conditions.In summary,we have established an optimized induction system that enhances the efficiency of h PGCLC differentiation by improving cellular resilience to hypoxic stress and apoptosis.展开更多
Hepatitis B virus infection remains a significant global health challenge,particularly in endemic regions like Vietnam.This article examines the groundbreaking study by Nguyen et al,which investigates the relationship...Hepatitis B virus infection remains a significant global health challenge,particularly in endemic regions like Vietnam.This article examines the groundbreaking study by Nguyen et al,which investigates the relationship between human leukocyte antigen-DP/DQ polymorphisms and hepatitis B virus-related liver disease progression.Through advanced multi-clustering analysis,the study reveals that the A-A-A haplotype(rs2856718-rs3077-rs9277535)provides protection against disease progression,while the G-G-G haplotype correlates with increased hepatocellular carcinoma susceptibility.The integration of machine learning approaches with genetic data offers promising avenues for refined disease prediction and personalized therapeutic strategies.This article discusses the implications for expanding study populations,implementing longitudinal cohort studies,and leveraging artificial intelligence for improved patient outcomes.展开更多
Viral reproductive tract infections(VRTIs)are a significant global health concern with severe consequences,including infertility,chronic pelvic pain,and increased risk of HIV transmission.Complex interplay between pat...Viral reproductive tract infections(VRTIs)are a significant global health concern with severe consequences,including infertility,chronic pelvic pain,and increased risk of HIV transmission.Complex interplay between pathogens and the host immune system plays a critical role in the pathogenesis and treatment of VRTIs.This review aims to provide a comprehensive overview of the multifaceted roles of the immune system in both contributing to and combating VRTIs.The review will also address the consequent perturbations in immune homeostasis and the implications for disease manifestation and progression.The interrelation between systemic immunity and local immune responses is discussed,providing insights into the challenges and breakthroughs in managing these infections.By providing a comprehensive overview of the mechanisms,implications,and therapeutic strategies associated with viral reproductive tract infections and immune dysfunction,this review also informs future research and clinical practice in this important area of reproductive health.展开更多
Background:The precise insertion of large DNA fragments(>3–5 kb)remains one of the key obstacles in establishment of genetically modified murine models.Methods:A 21 kb large DNA fragment containing three tandemly ...Background:The precise insertion of large DNA fragments(>3–5 kb)remains one of the key obstacles in establishment of genetically modified murine models.Methods:A 21 kb large DNA fragment containing three tandemly linked copies of the human HRAS gene was inserted into the genome of C57BL/6J mouse,generating a mouse model designated as KI.C57-ras(or named NF-h HRAS).Whole-genome sequencing and Sanger sequencing were utilized to it confirm precise insertion and copy number.The stability of transgene expression among different generations was verified from multiple aspects using by digital PCR,western blot and DNA sequencing.To assess tumor susceptibility in the mouse model,N-Nitroso-N-methylurea(MNU)was administered at a dosage of 75 mg/kg.Histopathological examinations were conducted using hematoxylin and eosin(H&E)staining.Results:The HRAS DNA fragment was inserted into mouse chromosome 15E1 site,locating between 80623202 bp and 80625020 bp.NF-h HRAS mice exhibited stable inheritance and displayed consistent phenotypes across individuals.Moreover,this mouse model exhibited a high susceptibility to carcinogens.Upon administration of MNU the earliest mortality onset was earlier than that of wild-type littermates(day 65 vs.day 78 for male and day 56 vs.day 84 for female).Notably,100%of the NF-h HRAS transgenic mice developed tumors,with approximately 84%of male NF-h HRAS mice exhibiting specific tumor types,such as squamous cell carcinoma or squamous cell papilloma,which was consistent with the previously reported carcinogenic rasH2 mouse model.The types of tumors and the target organs exhibited diversity in NFh HRAS mice,while the spontaneous tumor incidence remained low(1/50).Conclusions:The NF-h HRAS mice demonstrated excellent genetic stability,a reproducible phenotype,and high susceptibility to carcinogens,indicating their potential utility in non-clinical safety evaluations of drugs as per the S1B guidelines issued by the ICH(The International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use).展开更多
基金This work was supported by the National Natural Science Foundation of China(No. 39470293).
文摘Objective To express human B7. 2 extracellular domain with prokaryote expression system and to evaluate its biological activity in vitro. Methods PCR was used to amplify the extracellular region of human B7. 2 which contained both the IgV and IgC domains. The recombinant PGEX-4T-3/hB7. 2 (IgV+C) was obtained by cloning the PCR product into a prokaryote expression plasmid PGEX-4T-3 and was transformed into the host strain of DH5-a. Tke fusion protein consisted of GST and hB7. 2(IgV+C) was identified by SDS-PAGE and Western blotting. T cell activation was observed by exposing purified T lymphocytes to the fusion protein and [3H]-TdR incorporation with the presence of the first signal imitated hy anti-CD3 antibody. Results The fusion protein GST-hB7. 2 (IgV+ C) was produced and detected in inclusive body form from engineered bacterial cells. With the first signal existed,T lymphocytes proliferated when it was co-stimulated by the fusion protein. Conclusion These results indicated that the functional human B7. 2(IgV+C) fusion protein can be produced in bacterial cells and the fusion protein displays the co-stimulatory activity in T lymphocytes activation.
基金supported by the National Natural Science Foundation (NSFC) Distinguished Young Scholar(32425053)the National KeyR&D Program of China(2023YFC2307500)+1 种基金the NSFC(32370166)the Beijing Research Center for Respiratory Infectious Diseases(BJRID2025-007).
文摘Since 2021,clade 2.3.4.4b H5N1 highly pathogenic avian influenza viruses(AIVs)have become the dominant strain responsible for the ongoing panzootics in birds worldwide[1],causing unprecedented deathsofpoultry and wild birds(https://wahis.-woah.org).Moreover,the H5N1 viruses have caused increasing infections in domestic and wild mammals[2].
文摘BACKGROUND Prevalence of the main rheumatic diseases in the Republic of Sakha(Yakutia)[RS(Y)],one of the regions of the Russian Federation,differs from the other regions of the Russian Federation due to its ethnic and geographic features.Knowledge regarding the prevalence and structure of juvenile idiopathic arthritis(JIA)allows us to shape the work of the pediatric rheumatology service in the region correctly,and optimize the healthcare system and the need for medica-tions.AIM To describe the epidemiological,demographic,clinical,and laboratory characteristics of children with JIA in the RS(Y)and evaluate the main outcomes.METHODS This retrospective cohort study assessed all the data from the medical histories of the patients(n=225)diagnosed with JIA(2016-2023)in the Cardiorheumatology Department of the M.E.Nikolaev National Center of Medicine.Pearson'sχ²test,Fisher's exact test,Mann–Whitney and Kruskal-Wallis tests were used for statistical analyses.RESULTS The ethnic prevalence of JIA is higher in Sakha than in Russian children at 110.1 per 100000 children and 69.4 per 100000 children,respectively.The prevalence of JIA among boys and girls in Sakha was similar,unlike in Russians,where the number of girls predominated.The JIA categories were as follows:(1)Systemic arthritis:3.5%;(2)Oligoarthritis(persistent and extended):33.8%;(3)Rheumatoid factor(RF)(+)polyarthritis:0.9%;(4)RF(-)polyarthritis:14.7%;(5)Enthesitis-related arthritis(ERA):44%;and(6)Psoriatic arthritis:3.1%.Prevalence of the ERA category was 4.4 times higher in Sakha children,but the prevalence of systemic arthritis was 2.9 times lower compared to Russians(P=0.0005).The frequency of uveitis was 10.2%,and the frequency of human leukocyte antigen(HLA)B27 was 39.6%in JIA children.Biologic treatment was received by 40.4%of JIA children and 45.3%achieved remission.CONCLUSION Higher JIA prevalence,male and ERA predominance,related to a higher frequency of HLA B27 are typical in RS(Y).These data might improve the pediatric rheumatology health service.
基金funded by the National Key R&D Program(Grant Nos.2022YFC2702800 and 2021YFC2700302 to Y.Y.)the National Natural Science Foundation of China(Grant Nos.82122025 to Y.Y.,82221005 to J.S.,and 82201763 to L.L.)。
文摘Primordial germ cells(PGCs),the precursors of oocytes and spermatozoa,are highly pluripotent.In recent years,the in vitro induction of human primordial germ cell-like cells(h PGCLCs)has advanced significantly.However,the stability and efficacy of obtaining h PGCLCs in vitro still require improvement.In the current study,we identified a novel induction system using Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12(DMEM/F-12)as the basal medium,supplemented with B27 and N2(referred to as N2B27)in combination with four cytokines:bone morphogenetic protein 4,stem cell factor,epidermal growth factor,and leukemia inhibitory factor.The h PGCLCs induced under these conditions closely resembled PGCs from 4-to 5-week-old embryos at the transcriptomic level.Compared with traditional GK15(GMEM supplemented with 15%Knockout?Serum Replacement)-based induction conditions,the N2B27 system significantly increased the speed and efficacy of h PGCLC induction.RNA sequencing analysis revealed that this improvement was due to an increased cellular capacity to cope with hypoxic stress and avoid apoptosis.The N2B27 medium promoted enhanced mitochondrial activity,enabling cells to better manage hypoxic stress while reducing the production of reactive oxygen species.Moreover,through gradient concentration experiments,we demonstrated that the addition of the common antioxidant N-acetyl-L-cysteine at an optimized concentration further enhanced the efficiency of PGCLC induction under GK15 conditions.In summary,we have established an optimized induction system that enhances the efficiency of h PGCLC differentiation by improving cellular resilience to hypoxic stress and apoptosis.
基金Supported by National Natural Science Foundation of China,No.82170406 and No.81970238.
文摘Hepatitis B virus infection remains a significant global health challenge,particularly in endemic regions like Vietnam.This article examines the groundbreaking study by Nguyen et al,which investigates the relationship between human leukocyte antigen-DP/DQ polymorphisms and hepatitis B virus-related liver disease progression.Through advanced multi-clustering analysis,the study reveals that the A-A-A haplotype(rs2856718-rs3077-rs9277535)provides protection against disease progression,while the G-G-G haplotype correlates with increased hepatocellular carcinoma susceptibility.The integration of machine learning approaches with genetic data offers promising avenues for refined disease prediction and personalized therapeutic strategies.This article discusses the implications for expanding study populations,implementing longitudinal cohort studies,and leveraging artificial intelligence for improved patient outcomes.
基金supported by the Universiti Malaya ArtScience Fellowship Grant(UMG004-2024IAS)UM Community Engagement Grant 2023(Project RUU002-23KS).
文摘Viral reproductive tract infections(VRTIs)are a significant global health concern with severe consequences,including infertility,chronic pelvic pain,and increased risk of HIV transmission.Complex interplay between pathogens and the host immune system plays a critical role in the pathogenesis and treatment of VRTIs.This review aims to provide a comprehensive overview of the multifaceted roles of the immune system in both contributing to and combating VRTIs.The review will also address the consequent perturbations in immune homeostasis and the implications for disease manifestation and progression.The interrelation between systemic immunity and local immune responses is discussed,providing insights into the challenges and breakthroughs in managing these infections.By providing a comprehensive overview of the mechanisms,implications,and therapeutic strategies associated with viral reproductive tract infections and immune dysfunction,this review also informs future research and clinical practice in this important area of reproductive health.
基金National Key R&D Program of China,Grant/Award Number:2023YFC3402000National Institutes for Food and Drug Control,State Key Laboratory of Drug Regulatory Science,Grant/Award Number:2023SKLDRS0124。
文摘Background:The precise insertion of large DNA fragments(>3–5 kb)remains one of the key obstacles in establishment of genetically modified murine models.Methods:A 21 kb large DNA fragment containing three tandemly linked copies of the human HRAS gene was inserted into the genome of C57BL/6J mouse,generating a mouse model designated as KI.C57-ras(or named NF-h HRAS).Whole-genome sequencing and Sanger sequencing were utilized to it confirm precise insertion and copy number.The stability of transgene expression among different generations was verified from multiple aspects using by digital PCR,western blot and DNA sequencing.To assess tumor susceptibility in the mouse model,N-Nitroso-N-methylurea(MNU)was administered at a dosage of 75 mg/kg.Histopathological examinations were conducted using hematoxylin and eosin(H&E)staining.Results:The HRAS DNA fragment was inserted into mouse chromosome 15E1 site,locating between 80623202 bp and 80625020 bp.NF-h HRAS mice exhibited stable inheritance and displayed consistent phenotypes across individuals.Moreover,this mouse model exhibited a high susceptibility to carcinogens.Upon administration of MNU the earliest mortality onset was earlier than that of wild-type littermates(day 65 vs.day 78 for male and day 56 vs.day 84 for female).Notably,100%of the NF-h HRAS transgenic mice developed tumors,with approximately 84%of male NF-h HRAS mice exhibiting specific tumor types,such as squamous cell carcinoma or squamous cell papilloma,which was consistent with the previously reported carcinogenic rasH2 mouse model.The types of tumors and the target organs exhibited diversity in NFh HRAS mice,while the spontaneous tumor incidence remained low(1/50).Conclusions:The NF-h HRAS mice demonstrated excellent genetic stability,a reproducible phenotype,and high susceptibility to carcinogens,indicating their potential utility in non-clinical safety evaluations of drugs as per the S1B guidelines issued by the ICH(The International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use).
