Objective To explore the application effect of"5E"teaching mode combined with deep learning charac-teristics in residential training teaching of general subjects.Methods From July to December 2024,the"5E"teaching mode combined with the characteristics of deep learning was implemented in 30 general practitioners being trained in three standardized training bases of residents in Qiqihar City,and the scores of self-learning ability and critical thinking ability of students before and after the implementation of teaching were tested by paired t-test.Re-sults After teaching,the scores of self-learning ability and critical thinking ability of the general training students were higher than those before teaching,and the differences were statistically significant(P＜0.05).Conclusion"5E"teaching mode can improve the learning effect and clinical practice ability of general training students.

Objective:To obtain a polyvalent single-chain variable fragment（scFv）against West Nile virus through PEGylation in order to improve its antigen-binding ability and neutralizing activity.Methods:A scFv carrying a C-terminal cysteine residue（scFvC）was constructed by introducing Cys into the C-terminal of scFv against West Nile virus. Then the multimerization of scFvC was achieved by targeting the thiol group of Cys with maleimide-activated polyethylene glycol. ELISA was used to detect the antigen-binding activity of the multivalent scFvC. Pseudovirus-based neutralization assay was used to evaluate the neutralizing activity of the multivalent scFvC in vitro. One-way analysis of variance was used for statistical analysis. Results:The PEGylated scFvC multimers showed higher antigen-binding ability than the monomeric scFvC. In the pseudovirus-based neutralization assay，both monomeric scFvC and PEGylated scFvC multimers showed good neutralizing activity compared with the control group（ P<0.000 1）. Moreover，the PEGylated scFvC multimers showed a more effective ability to block the pseudovirus infection in target cells（ P<0.05），suggesting that the PEGylated scFvC multimers could enhance their function in vitro through avidity effect. Conclusion:In this study，a scFvC targeting West Nile virus is successfully constructed and its polyvalent form is generated through PEGylation，which improves the antigen-binding and neutralizing activity of the parental scFv.

Monkeypox,caused by the monkeypox virus(MPXV),is a zoonotic infectious disease characterized clinically by fever,rash,and lymphadenopathy.In September 2024,the outbreak of a novel MPXV Clade Ib variant in the Democratic Republic of the Congo was designated by the World Health Organization as a public health emergency of international concern,highlighting deficiencies in current prevention and control systems.Although modified attenuated smallpox vaccines exhibit cross-protective efficacy against MPXV infection,their adverse effects still limit clinical applications.Further-more,the therapeutic efficacy of tecovirimat—the FDA emergency-authorized antiviral drug for Smallpox—is limited.Recent studies have shown that tecovirimat does not significantly improve lesion healing time or reduce overall mortality in patients infected with MPXV CladeⅠ.Monoclonal antibodies,which neutralize viral activity by targeting critical membrane proteins,have emerged as a pivotal strategy to overcome current therapeutic bottlenecks by integrating therapeutic,diagnostic and prophylactic functions.This article reviews the epidemiological and virological characteristics of MPXV,along with functional characteristics of MPXV major membrane proteins,including A29L,A35R,B6R,E8L,M1R and H3L,focusing on the research progress in monoclonal antibodies against these key targets.Their protective effects in vitro/in vivo are explored while such as strategies monoclonal antibody combina-tion therapy are recommended to enhance efficacy and overcome drug resistance.

Intricate and complex interactions between tumor cells and tumor microenvironment(TME)are essential driving factors in tumor development and metastasis.In recent years,modulation of TME to reverse tumor progression has garnered significant attention as a therapeutic strategy.For precise and targeted tumor treatment,there is an urgent need for accurate and effective TME tar-get molecules and relevant prediction model.Fibronectin,a protein with multiple domains in extracellular matrix(ECM),plays a cru-cial role in physiological and pathological processes.Particularly,Fibronectin extra domain B(FN-EDB)in fibronectin is pivotal in regulating interaction between ECM and cells.During active tissue remodeling processes,especially in angiogenesis,FN-EDB expres-sion tends to increase.Studies have indicated that FN-EDB is involved in regulating various key aspects of tumorigenesis,including promoting tumor cell invasion and metastasis,influencing development of immune evasion mechanisms,enhancing tumor angiogene-sis,and contributing to development of chemoresistance.Therefore,targeting FN-EDB as a potential therapeutic target for cancer treat-ment and developing relevant therapeutic drugs hold significant importance and promising clinical prospects.This article reviews roles of FN-EDB in TME in recent years and research progress on tumor immunotherapy targeting this molecule,with aim of providing new insights and strategies for cancer treatment.

Objective:To obtain fucose-free anti-West Nile virus nonstructural protein 1 (NS1) antibody and evaluate its antibody-dependent cell-mediated cytotoxicity (ADCC).Methods:The guanosine diphosphate-fucose transporter SLC35C1 in CHO cells was knocked out using CRISPR/Cas9 gene editing technology to obtain the fucose-free cell line CHO SLC35C1 -/-. CHO SLC35C1 -/- cells were used to produce fucose-free anti-West Nile virus NS1 antibodies. The binding abilities of the antibodies to the target antigen of West Nile virus NS1 protein and the human high-affinity IgG Fc receptor hFcγRⅠ (hCD64) were detected by ELISA and flow cytometry, respectively. The ADCC activity of the antibodies was detected by ADCC reporter gene assay. One-way analysis of variance was used for statistical analysis. Results:CHO SLC35C1 -/- cells expressed green fluorescent protein but not Lens culinaris agglutinin. The anti-West Nile virus NS1 antibodies produced by CHO SLC35C1 -/- cells with a fucose content of 0.22% could bind to West Nile virus NS1 protein in a concentration-dependent manner. Compared with the wild-type antibodies, the fucose-free anti-West Nile virus NS1 antibodies showed a stronger binding ability to hFcγRⅠ(hCD64), as indicated by a significant increase in fluorescence intensity. The ADCC reporter gene assay showed that the fucose-free anti-West Nile virus NS1 antibodies had increased activity as compared with the wild-type antibodies ( P<0.001). Conclusion:The fucose-free anti-West Nile virus NS1 antibodies may be used to protect against West Nile virus infection.

Monkeypox,caused by the monkeypox virus(MPXV),is a zoonotic infectious disease characterized clinically by fever,rash,and lymphadenopathy.In September 2024,the outbreak of a novel MPXV Clade Ib variant in the Democratic Republic of the Congo was designated by the World Health Organization as a public health emergency of international concern,highlighting deficiencies in current prevention and control systems.Although modified attenuated smallpox vaccines exhibit cross-protective efficacy against MPXV infection,their adverse effects still limit clinical applications.Further-more,the therapeutic efficacy of tecovirimat—the FDA emergency-authorized antiviral drug for Smallpox—is limited.Recent studies have shown that tecovirimat does not significantly improve lesion healing time or reduce overall mortality in patients infected with MPXV CladeⅠ.Monoclonal antibodies,which neutralize viral activity by targeting critical membrane proteins,have emerged as a pivotal strategy to overcome current therapeutic bottlenecks by integrating therapeutic,diagnostic and prophylactic functions.This article reviews the epidemiological and virological characteristics of MPXV,along with functional characteristics of MPXV major membrane proteins,including A29L,A35R,B6R,E8L,M1R and H3L,focusing on the research progress in monoclonal antibodies against these key targets.Their protective effects in vitro/in vivo are explored while such as strategies monoclonal antibody combina-tion therapy are recommended to enhance efficacy and overcome drug resistance.

Objective:To obtain fucose-free anti-West Nile virus nonstructural protein 1 (NS1) antibody and evaluate its antibody-dependent cell-mediated cytotoxicity (ADCC).Methods:The guanosine diphosphate-fucose transporter SLC35C1 in CHO cells was knocked out using CRISPR/Cas9 gene editing technology to obtain the fucose-free cell line CHO SLC35C1 -/-. CHO SLC35C1 -/- cells were used to produce fucose-free anti-West Nile virus NS1 antibodies. The binding abilities of the antibodies to the target antigen of West Nile virus NS1 protein and the human high-affinity IgG Fc receptor hFcγRⅠ (hCD64) were detected by ELISA and flow cytometry, respectively. The ADCC activity of the antibodies was detected by ADCC reporter gene assay. One-way analysis of variance was used for statistical analysis. Results:CHO SLC35C1 -/- cells expressed green fluorescent protein but not Lens culinaris agglutinin. The anti-West Nile virus NS1 antibodies produced by CHO SLC35C1 -/- cells with a fucose content of 0.22% could bind to West Nile virus NS1 protein in a concentration-dependent manner. Compared with the wild-type antibodies, the fucose-free anti-West Nile virus NS1 antibodies showed a stronger binding ability to hFcγRⅠ(hCD64), as indicated by a significant increase in fluorescence intensity. The ADCC reporter gene assay showed that the fucose-free anti-West Nile virus NS1 antibodies had increased activity as compared with the wild-type antibodies ( P<0.001). Conclusion:The fucose-free anti-West Nile virus NS1 antibodies may be used to protect against West Nile virus infection.

Objective To explore the application effect of"5E"teaching mode combined with deep learning charac-teristics in residential training teaching of general subjects.Methods From July to December 2024,the"5E"teaching mode combined with the characteristics of deep learning was implemented in 30 general practitioners being trained in three standardized training bases of residents in Qiqihar City,and the scores of self-learning ability and critical thinking ability of students before and after the implementation of teaching were tested by paired t-test.Re-sults After teaching,the scores of self-learning ability and critical thinking ability of the general training students were higher than those before teaching,and the differences were statistically significant(P＜0.05).Conclusion"5E"teaching mode can improve the learning effect and clinical practice ability of general training students.

Intricate and complex interactions between tumor cells and tumor microenvironment(TME)are essential driving factors in tumor development and metastasis.In recent years,modulation of TME to reverse tumor progression has garnered significant attention as a therapeutic strategy.For precise and targeted tumor treatment,there is an urgent need for accurate and effective TME tar-get molecules and relevant prediction model.Fibronectin,a protein with multiple domains in extracellular matrix(ECM),plays a cru-cial role in physiological and pathological processes.Particularly,Fibronectin extra domain B(FN-EDB)in fibronectin is pivotal in regulating interaction between ECM and cells.During active tissue remodeling processes,especially in angiogenesis,FN-EDB expres-sion tends to increase.Studies have indicated that FN-EDB is involved in regulating various key aspects of tumorigenesis,including promoting tumor cell invasion and metastasis,influencing development of immune evasion mechanisms,enhancing tumor angiogene-sis,and contributing to development of chemoresistance.Therefore,targeting FN-EDB as a potential therapeutic target for cancer treat-ment and developing relevant therapeutic drugs hold significant importance and promising clinical prospects.This article reviews roles of FN-EDB in TME in recent years and research progress on tumor immunotherapy targeting this molecule,with aim of providing new insights and strategies for cancer treatment.

Objective:To obtain a polyvalent single-chain variable fragment（scFv）against West Nile virus through PEGylation in order to improve its antigen-binding ability and neutralizing activity.Methods:A scFv carrying a C-terminal cysteine residue（scFvC）was constructed by introducing Cys into the C-terminal of scFv against West Nile virus. Then the multimerization of scFvC was achieved by targeting the thiol group of Cys with maleimide-activated polyethylene glycol. ELISA was used to detect the antigen-binding activity of the multivalent scFvC. Pseudovirus-based neutralization assay was used to evaluate the neutralizing activity of the multivalent scFvC in vitro. One-way analysis of variance was used for statistical analysis. Results:The PEGylated scFvC multimers showed higher antigen-binding ability than the monomeric scFvC. In the pseudovirus-based neutralization assay，both monomeric scFvC and PEGylated scFvC multimers showed good neutralizing activity compared with the control group（ P<0.000 1）. Moreover，the PEGylated scFvC multimers showed a more effective ability to block the pseudovirus infection in target cells（ P<0.05），suggesting that the PEGylated scFvC multimers could enhance their function in vitro through avidity effect. Conclusion:In this study，a scFvC targeting West Nile virus is successfully constructed and its polyvalent form is generated through PEGylation，which improves the antigen-binding and neutralizing activity of the parental scFv.