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.

ObjectiveTo investigate the optimal ratio of goat horn replacing antelope horn in Fufang Lingjiao Jiangya pills and the blood pressure-lowering mechanism of this medicine. MethodsThe blood pressure-lowering efficacy of Fufang Lingjiao Jiangya pills with varying proportions of goat horn replacing antelope horn was evaluated on spontaneous hypertensive rats （SHR）. In this experiment， 50 SHR rats were randomly grouped as follows： model （n=8）， captopril （0.01 g·kg^-1） （n=6）， low-dose blank Fufang Lingjiao Jiangya pills （0.342 g·kg^-1） （n=6）， high-dose blank Fufang Lingjiao Jiangya pills （0.684 g·kg^-1） （n=6）， low-dose antelope horn-containing Fufang Lingjiao Jiangya pills （0.378 g·kg^-1） （n=6）， high-dose antelope horn-containing Fufang Lingjiao Jiangya pills （0.756 g·kg^-1） （n=6）， low-dose goat horn-containing Fufang Lingjiao Jiangya pills （0.378 g·kg^-1） （n=6）， and high-dose goat horn-containing Fufang Lingjiao Jiangya pills （0.756 g·kg^-1） （n=6）. Additionally， 8 WKY rats were used as the normal group. Drugs were administered by gavage for 4 weeks while an equal volume of distilled water was administered for the normal and model groups. Blood pressure was measured before administration， 3 h post administration， and biweekly thereafter. In the experiment for Fufang Lingjiao Jiangya pills with goat horn replacing antelope horn in different proportions， 48 SHR rats were randomly grouped as follows： model， blank Fufang Lingjiao Jiangya pills （0.684 g·kg^-1）， antelope horn-containing Fufang Lingjiao Jiangya pills （0.756 g·kg^-1）， 2× goat horn-containing Fufang Lingjiao Jiangya pills （0.824 g·kg^-1）， 4× goat horn Fufang Lingjiao Jiangya pills （0.969 g·kg^-1）， and 6× goat horn Fufang Lingjiao Jiangya pills （1.112 g·kg^-1）. The normal group included 8 WKY rats， and the normal group and model group received an equal volume of distilled water. The treatment lasted for 2 weeks， and blood pressure was recorded at various time points （pre-administration， 3 h post administration， and on days 4， 7， 10， and 14 of administration）. Serum levels of angiotensin-converting enzyme （ACE）， angiotensin Ⅱ（Ang Ⅱ）， renin， and interleukin-6 （IL-6） were measured by enzyme-linked immunosorbent assay. Histopathological changes in the heart， kidney， and thoracic aorta were observed by hematoxylin-eosin staining. The protein levels of ACE2， angiotensin Ⅱ type 1 receptor （AT1R）， and angiotensinogen （AGT） in the kidney tissue were determined by Western blot， while the expression of nuclear factor （NF）-κB p65 and Toll-like receptor 4 （TLR4） in the thoracic aorta tissue was assessed by immunohistochemistry. ResultsCompared with the model group， all treatment groups showed lowered blood pressure （P<0.05， P<0.01）， and the 6× goat horn-containing Fufang Lingjiao Jiangya pills group showed consistent blood pressure-lowering effect with the antelope horn-containing Fufang Lingjiao Jiangya pills group. Compared with the normal group， the model group showed elevated serum levels of ACE， Ang Ⅱ， renin， and IL-6， while the elevations were declined in the Fufang Lingjiao Jiangya pills groups （P<0.05， P<0.01）. Pathological changes in the heart， kidney， and thoracic aorta were alleviated in all the treatment groups， with the 6× goat horn- and antelope horn-containing Fufang Lingjiao Jiangya pills groups exhibited the best effect. Western blot and immunohistochemistry results showed that all the treatment groups exhibited down-regulated protein levels of AT1R， AGT， NF-κB p65， and TLR4 and up-regulated protein levels of ACE2 （P<0.05， P<0.01） compared with model group， with the 6×goat horn- and antelope horn-containing Fufang Lingjiao Jiangya pills groups showcasing the best effect. ConclusionReplacing antelope horn with 6×goat horn in Fufang Lingjiao Jiangya pills can achieve consistent blood pressure-lowering effect with the original prescription. The prescription may exert the effect by inhibiting the renin-angiotensin-aldosterone system （RAAS） and TLR4/NF-κB signaling pathways.

Objective To identify mechanisms regulating disease progression in rat models of pulmonary arterial hy-pertension(PAH).Methods Rat PAH models were established using subcutaneous monocrotaline(MCT)injec-tion and the SU5416/hypoxia(SU/Hx)method.Transcriptomic sequencing of lung tissues was performed to identify gene expression and pathway alterations in PAH rats,followed by a comparative analysis with transcriptomic data of patients with idiopathic pulmonary arterial hypertension(IPAH)in NCBI database.Results Inflammatory-related genes such as CXCL9,CCL24,and SECTM1 were upregulated in both PAH rat models and IPAH patient lungs,while genes such as DGKG and DOCK9 were downregulated(P＜0.05).Pathways related to endothelial cell proliferation regulation and extracellular matrix(ECM)remodeling were significantly upregulated(P＜0.05).Conclusions The imbalance in endothelial cell proliferation and abnormal ECM remodeling may collectively contribute to PAH pathogenesis.Further exploration of these signaling pathways may provide deep in-sights for early diagnosis and targeted therapy of PAH.

Objective This scoping review systematically examines current evidence on prehabilitation interventions for patients undergoing hepatocellular carcinoma(HCC)surgery.The study aims to establish an evidence-based foundation for optimizing perioperative care in clinical nursing practice and inform the development of standardized prehabilitation protocols for future research.Methods According to the research method of scope review,Cochrane Library,PubMed,Embase,Web of Science,China Biomedical Literature Database,China Knowledge Network,Wanfang Database,and VIP Database were searched.The included literature was screened,summarized and analyzed.The literature search covered the period from the establishment of the database up to 30 November 2024.Totally 2 professionally trained researchers independently screened,extracted,and analyzed the included studies in accordance with the predefined inclusion and exclusion criteria.Results A total of 16 papers were included,including 11 randomized controlled studies(RCTs),4 retrospective studies and 1 class-experimental study.Prehabilitation is systematically implemented through 3 core modalities(nutritional optimization,structured exercise training,and psychological interventions)to enhance patients' physiological and psychological readiness for surgery,thereby improving operative outcomes and facilitating postoperative recovery.Conclusion Adopting a series of prehabilitation measures for patients with HCC undergoing elective surgery can improve the physical and psychological status of patients with HCC,enhance postoperative quality of life,and thus optimize clinical outcomes.Future studies should focus on exploring the optimal combination of prehabilitation strategies,formulating precise programmes based on individualized differences,strengthening patient compliance monitoring,and realizing efficient management with the help of digital technology.

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 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 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.

Objective This scoping review systematically examines current evidence on prehabilitation interventions for patients undergoing hepatocellular carcinoma(HCC)surgery.The study aims to establish an evidence-based foundation for optimizing perioperative care in clinical nursing practice and inform the development of standardized prehabilitation protocols for future research.Methods According to the research method of scope review,Cochrane Library,PubMed,Embase,Web of Science,China Biomedical Literature Database,China Knowledge Network,Wanfang Database,and VIP Database were searched.The included literature was screened,summarized and analyzed.The literature search covered the period from the establishment of the database up to 30 November 2024.Totally 2 professionally trained researchers independently screened,extracted,and analyzed the included studies in accordance with the predefined inclusion and exclusion criteria.Results A total of 16 papers were included,including 11 randomized controlled studies(RCTs),4 retrospective studies and 1 class-experimental study.Prehabilitation is systematically implemented through 3 core modalities(nutritional optimization,structured exercise training,and psychological interventions)to enhance patients' physiological and psychological readiness for surgery,thereby improving operative outcomes and facilitating postoperative recovery.Conclusion Adopting a series of prehabilitation measures for patients with HCC undergoing elective surgery can improve the physical and psychological status of patients with HCC,enhance postoperative quality of life,and thus optimize clinical outcomes.Future studies should focus on exploring the optimal combination of prehabilitation strategies,formulating precise programmes based on individualized differences,strengthening patient compliance monitoring,and realizing efficient management with the help of digital technology.

Objective:To explore the latent profile and characteristics of social isolation in patients with hematologic malignancy, and to analyze its related influencing factors, and to provide reference for improving social phobia disorder in different patients and implementing targeted intervention.Methods:This study was a cross-sectional survey. A convenient sampling method was used to select hematologic malignancy patients who were treated in Qilu Hospital of Shandong University from January 2022 to January 2023. General information questionnaire, the General Alienation Scale (GAS), and the Social Support Rating Scale(SSRS) were used for investigation. Latent profile was analyzed using the categories of social isolation in patients with hematologic malignancy, and univariate and multinomial logistic regression analysis were used to analyze relevant influencing factors.Results:A total of 195 survey subjects were included, of which 108 males and 87 females, aged (49.78 ± 13.52) years. The scores of GAS and SSRS were (43.21 ± 6.09) and (42.52 ± 6.77) respectively. The social isolation in patients with hematologic malignancy could be divided into 3 latent profiles, namely low-risk isolation 15.4% (30/195), medium-risk isolation 68.2%(133/195), and high-risk isolation16.4% (32/195). Multinomial Logistic regression analysis showed that age ( OR=0.941, 95% CI 0.894-0.990), percapita monthly income of families ( OR=0.050, 95% CI 0.004-0.657), primary caregivers (parents) ( OR=0.025, 95% CI 0.003-0.227), place of residence (town)( OR=0.170, 95% CI 0.039-0.749), disease type (leukemia) ( OR=15.610, 95% CI 2.973-81.979), disease type(lymphoma) ( OR=10.986, 95% CI 2.032-59.413) were the influencing factors of medium-risk isolation (all P<0.05). Age ( OR=0.933, 95% CI 0.880-0.988), percapita monthly income of families ( OR=0.029, 95% CI 0.002-0.525), primary caregivers (parents) ( OR=0.076, 95% CI 0.006-0.900), disease type (leukemia)( OR=19.257, 95% CI 2.580-143.723), disease type (lymphoma)( OR=9.952, 95% CI 1.290-76.763), social support ( OR=0.877, 95% CI 0.786-0.980) were the influencing factors of high-risk isolation (all P<0.05). Conclusions:The social isolation among patients with hematologic malignancy had apparent classification characteristics. It could be divided into three potential profiles. It is suggested that medical staff should take targeted social and psychological support based on different types of patients, improve their psychological and social outcomes, and utilize existing resources to implement intervention measures to help them adapt and return to society.

Since emergence,the influenza virus has triggered numerous global pandemics and claimed more than ten million lives.This virus poses not only a severe threat to human life and health,but significant challenges to global economy and public health.The research on highly pathogenic influenza virus strains(such as H5 and H7)necessitates the use of biosafety level 3 laboratories,which significantly escalates the experimental risks and costs.The pseudovirus technology,as a relatively safe and effective research technique,has been applied in the studies of various high-risk viruses.Thanks to constant research and refinement,the pseudovirus technology for the influenza virus currently boasts such advantages as user-friendliness and good safety,and has been extensively used antibody neutralization and screening of antiviral drugs.This article reviews the research progress in the pseudovirus technology for the influenza virus.