Moloney leukemia virus 10(MOV10)gene knockout(MKO)mouse neuro 2a(N2a)cell lines was constructed by CRISPR/Cas9(clustered regularly interspaced short palindromic repeat/CRISPR-associated protein 9)gene editing technology.First,a recombinant plasmid pMD18T-U6-sgRNA expressing MOV10 gene-specific guide RNA(sgRNA)was constructed,and then pMD18T-U6-sgRNA and pMJ920-Cas9-eGFP were co-transfected into N2a.The results showed that the MKO N2a cell line had normal cell activity and cell proliferation ability.The infection test of the MKO N2a cell line was carried out using the rabies virus(RABV)and vesicular stomatitis virus(VSV)of the Rhabdoviridae family.The results showed that the replication level of the Rhabdoviridae virus in the MKO N2a cell line was significantly enhanced.The results showed that a MKO N2a cell line was successfully constructed in this study,which provided a preliminary basis for the exploration of the biological function and antiviral mechanism of MOV10 and the develop-ment of a recombinant viral vector vaccine with RABV/VSV as the vector.

Objective:To explore the application effect of BOPPPS (Bridge-in, Objectives, Pre-assessment, Participatory learning, Post-assessment, and Summary) combined with scenario simulation in cultivating the humanistic care ability of undergraduate nursing interns.Methods:Undergraduate nursing students who were interning at the First Affiliated Hospital of Zhengzhou University were selected by convenience sampling. The 46 interns from June to December 2023 were set as the control group, and the 46 interns from June to December 2024 were set as the experimental group. The control group adopted the traditional lecture-based teaching method, while the experimental group adopted BOPPPS combined with scenario simulation. Theoretical and practical scores, humanistic care ability, empathy ability, and critical thinking ability of the two groups were compared.Results:After the intervention, the theoretical and practical scores, as well as humanistic care ability, empathy ability, and critical thinking ability scores of the experimental group were all higher than those of the control group, and the differences were statistically significant ( P<0.05) . Conclusions:BOPPPS combined with scenario simulation plays a positive role in improving theoretical and practical scores, humanistic care ability, empathy ability, and critical thinking ability of undergraduate nursing interns, and it is worthy of further promotion and application.

To establish an indirect enzyme linked immunosorbent assay(ELISA)method for the detection of Zaire Ebola virus(ZEBOV)specific antibodies,the full-length of ZEBOV GP1 gene was amplified by PCR and cloned into pET-30a(+)vector to generate the pET-30a(+)-GP1 plasmid.After expressed in the E.coli expression system,the purified GP1 protein was used as coating antigen to establish the indirect ELISA method for detection of ZEBOV antibody.The con-ditions including concentration of coating antigen and serum dilution were determined by chess-board titration.Specificity,sensitivity,and reproducibility of the established ELISA detection meth-od were evaluated.GP1 protein was successfully prepared by prokaryotic expression,and was used as the coatingantigen for indirect ELISA.By optimizing the reaction conditions,the optimal concen-tration of the coating antigen was determined to be 0.5 g/L;the optimal dilution of serum was cal-culated to be 1∶3 200;the optimal dilution of enzyme-labeled secondary antibody was measured to be 1∶20 000.The established method exhibited excellent specificity,sensitivity,and reproducibili-ty.In the present study,the GP1 protein was successfully expressed in the E.coli expression sys-tem and the high purity GP1 protein was used as the coating protein to establish an indirect ELISA assay for ZEBOV antibody.This method is highly specific,sensitive,and reproducible,which provides technical support for the fur-ther study of the biological function of GP1 and the detection of ZEBOV antibody in serum.

Objective:To investigate the predictive value of the cancer antigen 125 (CA 125) elimination rate constant K (KELIM) score for no visible residual disease (R0) and prognosis in high-grade serous ovarian carcinoma (HGSOC) patients undergoing neoadjuvant chemotherapy (NACT)+interval debulking surgery (IDS). Methods:A retrospective analysis was conducted on 78 HGSOC patients treated with NACT+IDS at Beijing Hospital, from June 2014 to June 2024. The KELIM score was calculated, and its predictive value for R0 resection, chemotherapy response score (CRS), platinum-free interval (PFI), progression-free survival (PFS) time, and overall survival (OS) time was analyzed.Results:(1) The mean age at diagnosis was (61.9±9.9) years. The mean KELIM score was 1.1±0.4, with 44 patients having KELIM score≥1 and 34 having KELIM score <1. (2) Patients with KELIM score ≥1 had significantly higher rates of R0 resection (84% vs 56%; P=0.006), CRS3 grading (41% vs 0; P<0.001), and PFI ≥6 months (84% vs 53%; P=0.04) compared to those with KELIM score <1. Additionally, the median PFS time (18.7 vs 13.2 months; P<0.001) and OS time (34.8 vs 29.9 months; P=0.007) were significantly longer in the KELIM score ≥1 group. Chemosensitivity: patients with PFI <6 months had a significantly lower median KELIM score than those with PFI ≥6 months (0.8 vs 1.2; P=0.005). Surgical outcome: patients achieving R0 resection had a significantly higher median KELIM score than those without R0 (1.2 vs 0.7; P<0.001). (3) Univariate analysis identified non-R0 resection, CRS3 grading, lack of poly adenosine diphosphate ribose polymerase (PARP) inhibitor maintenance therapy, and KELIM score <1 as significant risk factors for OS time (all P<0.05). Multivariate analysis confirmed non-R0 resection ( HR=3.78,95% CI: 1.13-12.66; P=0.031), no PARP inhibitor maintenance ( HR=7.41,95% CI:1.82-30.15; P=0.005), and KELIM score <1 ( HR=5.14,95% CI:1.41-18.72; P=0.013) as independent risk factors for OS time. Conclusions:The KELIM score may serve as a predictive marker for chemosensitivity, R0 resection, PFS time, and OS time in HGSOC patients undergoing NACT+IDS. KELIM score<1 is an independent risk factor for OS.

Objective:To explore the risk factors of pneumoconiosis complicated with pulmonary tuberculosis, to construct a clinical prediction model for patients with pneumoconiosis complicated with pulmonary tuberculosis, and to provide a scientific basis for the prevention of pneumoconiosis complicated with pulmonary tuberculosis.Methods:In January 2024, a total of 232 patients with pneumoconiosis (including coal workers' pneumoconiosis and silicosis) who were treated in the Department of Respiratory and Critical Care Medicine of the Third People's Hospital of Xinjiang Uygur Autonomous Region (Xinjiang Uygur Autonomous Region Occupational Disease Hospital) from January 2022 to January 2023 were randomly selected as the study subjects. Collectted basic patient information and diagnostic data. Multivariate logistic regression analysis was used to screen the risk factors related to pneumoconiosis complicated with pulmonary tuberculosis. According to the results of multivariate logistic regression analysis, a nomogram was established, and the area under the receiver operating characteristic (ROC) curve (AUC), calibration curve and decision curve analysis (DCA) were used to evaluate the predictive ability.Results:Among the 232 patients with pneumoconiosis, 73 were complicated with pulmonary tuberculosis, accounting for 31.47% (73/232). Multivariate logistic regression analysis determined that dust exposure time, type of work, smoking history, and lung function level were all risk factors for pneumoconiosis complicated with tuberculosis ( OR=10.33, 95% CI=1.92~55.66, OR=5.43, 95% CI=1.91~15.44, OR=3.10, 95% CI=1.15~8.37, OR=4.00, 95% CI=1.62~9.87; P<0.05). The constructed nomogram model has good clinical applicability when the area under the receiver operating characteristic (ROC) curve is 0.77 [95% CI (0.69, 0.73) ], the calibration curve is close to the ideal diagonal, the absolute error between the simulation curve and the actual curve is 0.03, and the DCA decision curve shows that the probability threshold of the nomogram model is 1%-90%. Conclusion:The risk of pneumoconiosis complicated with tuberculosis is high, and the risk factors of dust exposure time, smoking history, type of work and lung function level are high. This nomogram model can be used to predict the risk of pulmonary tuberculosis in patients with pneumoconiosis, which is helpful for early intervention.

Moloney leukemia virus 10(MOV10)gene knockout(MKO)mouse neuro 2a(N2a)cell lines was constructed by CRISPR/Cas9(clustered regularly interspaced short palindromic repeat/CRISPR-associated protein 9)gene editing technology.First,a recombinant plasmid pMD18T-U6-sgRNA expressing MOV10 gene-specific guide RNA(sgRNA)was constructed,and then pMD18T-U6-sgRNA and pMJ920-Cas9-eGFP were co-transfected into N2a.The results showed that the MKO N2a cell line had normal cell activity and cell proliferation ability.The infection test of the MKO N2a cell line was carried out using the rabies virus(RABV)and vesicular stomatitis virus(VSV)of the Rhabdoviridae family.The results showed that the replication level of the Rhabdoviridae virus in the MKO N2a cell line was significantly enhanced.The results showed that a MKO N2a cell line was successfully constructed in this study,which provided a preliminary basis for the exploration of the biological function and antiviral mechanism of MOV10 and the develop-ment of a recombinant viral vector vaccine with RABV/VSV as the vector.

Objective:To investigate the predictive value of the cancer antigen 125 (CA 125) elimination rate constant K (KELIM) score for no visible residual disease (R0) and prognosis in high-grade serous ovarian carcinoma (HGSOC) patients undergoing neoadjuvant chemotherapy (NACT)+interval debulking surgery (IDS). Methods:A retrospective analysis was conducted on 78 HGSOC patients treated with NACT+IDS at Beijing Hospital, from June 2014 to June 2024. The KELIM score was calculated, and its predictive value for R0 resection, chemotherapy response score (CRS), platinum-free interval (PFI), progression-free survival (PFS) time, and overall survival (OS) time was analyzed.Results:(1) The mean age at diagnosis was (61.9±9.9) years. The mean KELIM score was 1.1±0.4, with 44 patients having KELIM score≥1 and 34 having KELIM score <1. (2) Patients with KELIM score ≥1 had significantly higher rates of R0 resection (84% vs 56%; P=0.006), CRS3 grading (41% vs 0; P<0.001), and PFI ≥6 months (84% vs 53%; P=0.04) compared to those with KELIM score <1. Additionally, the median PFS time (18.7 vs 13.2 months; P<0.001) and OS time (34.8 vs 29.9 months; P=0.007) were significantly longer in the KELIM score ≥1 group. Chemosensitivity: patients with PFI <6 months had a significantly lower median KELIM score than those with PFI ≥6 months (0.8 vs 1.2; P=0.005). Surgical outcome: patients achieving R0 resection had a significantly higher median KELIM score than those without R0 (1.2 vs 0.7; P<0.001). (3) Univariate analysis identified non-R0 resection, CRS3 grading, lack of poly adenosine diphosphate ribose polymerase (PARP) inhibitor maintenance therapy, and KELIM score <1 as significant risk factors for OS time (all P<0.05). Multivariate analysis confirmed non-R0 resection ( HR=3.78,95% CI: 1.13-12.66; P=0.031), no PARP inhibitor maintenance ( HR=7.41,95% CI:1.82-30.15; P=0.005), and KELIM score <1 ( HR=5.14,95% CI:1.41-18.72; P=0.013) as independent risk factors for OS time. Conclusions:The KELIM score may serve as a predictive marker for chemosensitivity, R0 resection, PFS time, and OS time in HGSOC patients undergoing NACT+IDS. KELIM score<1 is an independent risk factor for OS.

To establish an indirect enzyme linked immunosorbent assay(ELISA)method for the detection of Zaire Ebola virus(ZEBOV)specific antibodies,the full-length of ZEBOV GP1 gene was amplified by PCR and cloned into pET-30a(+)vector to generate the pET-30a(+)-GP1 plasmid.After expressed in the E.coli expression system,the purified GP1 protein was used as coating antigen to establish the indirect ELISA method for detection of ZEBOV antibody.The con-ditions including concentration of coating antigen and serum dilution were determined by chess-board titration.Specificity,sensitivity,and reproducibility of the established ELISA detection meth-od were evaluated.GP1 protein was successfully prepared by prokaryotic expression,and was used as the coatingantigen for indirect ELISA.By optimizing the reaction conditions,the optimal concen-tration of the coating antigen was determined to be 0.5 g/L;the optimal dilution of serum was cal-culated to be 1∶3 200;the optimal dilution of enzyme-labeled secondary antibody was measured to be 1∶20 000.The established method exhibited excellent specificity,sensitivity,and reproducibili-ty.In the present study,the GP1 protein was successfully expressed in the E.coli expression sys-tem and the high purity GP1 protein was used as the coating protein to establish an indirect ELISA assay for ZEBOV antibody.This method is highly specific,sensitive,and reproducible,which provides technical support for the fur-ther study of the biological function of GP1 and the detection of ZEBOV antibody in serum.

Objective:To explore the application effect of BOPPPS (Bridge-in, Objectives, Pre-assessment, Participatory learning, Post-assessment, and Summary) combined with scenario simulation in cultivating the humanistic care ability of undergraduate nursing interns.Methods:Undergraduate nursing students who were interning at the First Affiliated Hospital of Zhengzhou University were selected by convenience sampling. The 46 interns from June to December 2023 were set as the control group, and the 46 interns from June to December 2024 were set as the experimental group. The control group adopted the traditional lecture-based teaching method, while the experimental group adopted BOPPPS combined with scenario simulation. Theoretical and practical scores, humanistic care ability, empathy ability, and critical thinking ability of the two groups were compared.Results:After the intervention, the theoretical and practical scores, as well as humanistic care ability, empathy ability, and critical thinking ability scores of the experimental group were all higher than those of the control group, and the differences were statistically significant ( P<0.05) . Conclusions:BOPPPS combined with scenario simulation plays a positive role in improving theoretical and practical scores, humanistic care ability, empathy ability, and critical thinking ability of undergraduate nursing interns, and it is worthy of further promotion and application.

Objective:To investigate the anti-inflammatory activity of alcohol extract of Polyrhachis dives(PDAEs)against rheumatoid arthritis(RA)in vitro and in vivo.Methods:In vivo and in vitro anti-RA inflammatory response of PDAEs was investigated using a rat model of collagenous arthritis induced by bovine type Ⅱ collagen and an LPS-induced RAW264.7 cell inflammatory model.Results:PDAEs inhibited the polarization of M1 type macrophages in vivo and in vitro,reduced expressions of TNF-α,IL-1β,IL-6,iNOS,and promoted polarization of M2 type macrophages,enhanced expressions of anti-inflammatory cytokines,such as IL-10 and TGF-β,so as to achieve the anti-inflammatory effect.The experiments in vivo also showed that PDAEs had the immunomodulatory effect,the potential mechanism may be the regulation of Th17/Treg balance by regulating the expression of PD-1 and TGF-β,thus correcting the over-strong autoimmune response.Conclusion:PDAEs may reduce the inflammatory reaction of RA through anti-inflam-matory and immunomodulatory effects.