Objective To investigate the role and regulatory mechanism of LINC00657 in the progression of cervical cancer. Methods Bioinformatics analysis predicted potential binding sites between LINC00657 and miR-30a-5p and between miR-30a-5p and Skp2. These sites were verified by using RNA immunoprecipitation and dual-luciferase reporter experiments. LINC00657, miR-30a-5p, and Skp2 mRNA expression levels in cervical cancer tissues and cell lines were assessed by utilizing RT-qPCR. Western blot analysis was employed to examine the protein levels of Skp2 in cells and subcutaneous xenograft tumor models in nude mice. Immunohistochemistry was applied to analyze Skp2 expression in animal tissues. The cellular processes of cervical cancer cell lines were evaluated through CCK-8, scratch, and Transwell assays. Results LINC00657 and Skp2 presented binding sites for miR-30a-5p. In cervical cancer, LINC00657 and Skp2 showed high expression levels (P<0.05), whereas miR-30a-5p displayed low expression (P<0.05). Functional experiments demonstrated that linc00657 upregulates Skp2 expression, a process that is dependent on its sequestration of miR-30a-5p. Conclusion LINC00657 promoted the malignant progression of cervical cancer by upregulating Skp2 expression through specifically sequestering miR-30a-5p, thereby relieving its inhibitory effect on the target gene Skp2.

Objective: To evaluate the current situation of thermal environment in primary and secondary school classrooms during winter, and to analyze students thermal comfort needs, so as to provide a basis for improving classroom thermal environment. Methods: From December 16 to 26, 2024, a stratified cluster random sampling method was used to select 90 classrooms from 15 primary and secondary schools in centralized/air conditioned heating areas(Liaoning Province, Tianjin City, Shanghai City) and naturally ventilated areas(Anhui Province and Jiangxi Province)for on site environmental measurement. A questionnaire survey was conducted among 743 students. The differences between groups using the χ 2 test were compared. Based on actual measurement data, a predicted mean vote prepared percentage of dissatisfied (PMV-PPD) model for centralized/air conditioned classrooms and an adaptive model for naturally ventilated classrooms were established, and the thermal neutral temperature and comfort interval were calculated. Results: The average outdoor temperature during on site measurement was 4.00(0.20，7.00)℃. In classrooms with centralized or air conditioned heating systems, the measured average temperature was (19.33±2.59)℃, with a thermal comfort range of 20.35-25.35 ℃ and a thermal neutral temperature of 22.85 ℃. And 13.92% of students reported feeling cold, while 80.80% felt comfortable. In classrooms with natural ventilation, the measured average temperature was (12.26±1.83)℃, with a thermal neutral temperature of 19.67 ℃ and a thermal comfort range of 16.17-23.17 ℃. About 48.33% of students reported feeling cold, and 49.81 % felt comfortable.The results of univariate analysis showed that there were statistically significant differences in shoe thickness, temperature sensation, relative humidity sensation and wind speed sensation between centralized/air conditioned heating areas ( χ 2= 7.01 , 31.47, 13.57, 13.80,all P <0.05). There were also statistically significant differences in school stage for primary and secondary school students, body mass index, classroom location for seat, temperature sensation, relative humidity sensation and wind speed sensation between naturally ventilated areas ( χ 2=42.13, 11.13, 11.04, 60.39, 29.27, 38.46,all P <0.05). Conclusions There are differences in thermal environment and students subjective thermal comfort in primary and secondary schools under different ventilation modes in winter. The temperature standards for heated classrooms should be revised, and differentiated environmental regulation strategies should be adopted based on different ventilation methods to improve students health and comfort levels.

To prepare monoclonal antibody to the N protein of porcine reproductive and respiratory syndrome virus(PRRSV),BALB/c mice were immunized with the purified N protein of the PRRSV-2 strain expressed by prokaryotic expression system.Mouse splenocytes were fused with myeloma cells using hybridoma technique,hybridoma cells were identified by indirect ELISA method and indirect immunofluorescence assay(IFA),and positive hybridoma cells were screened for subclones using the limited dilution method.The results showed that a monoclonal antibody cell line 4A7 was successfully obtained,and the results of Western blot and IFA indicated that the monoclonal antibody could accurately recognize the N proteins of type 1 and type 2 PRRSV.Mean-while,the N protein gene was truncated and expressed by prokaryotic expression system,and the amino acid sequence of the B-cell antigenic epitope recognized by 4A7 was screened as 51EKPHF55 using Western blot.Comparison of epitope amino acids in the N protein gene sequences of different strains revealed that the antigenic epitope 51EKPHF55 recognized by the monoclonal antibody 4A7 has no amino acid difference in the sequences of three subtypes among the PRRSV-1 strains and nine lineages among the PRRSV-2 strains,indicating a high degree of conservation.The results of the study provide a foundation the development of PRRSV diagnostic kits and novel vaccines.

Objective:To investigate the feasibility of transcatheter edge-to-edge repair (TEER) using a short-clip strategy for patients with moderate-to-severe or greater degenerative mitral regurgitation caused by commissural prolapse.Methods:This retrospective study included patients with severe mitral regurgitation secondary to commissural prolapse who underwent TEER at the Second Affiliated Hospital of Zhejiang University School of Medicine between September 2022 and July 2024. Preoperative clinical and imaging data, intraoperative details, procedural outcomes, and 1-month postoperative follow-up results were collected.Results:A total of 19 patients were enrolled, aged (74.1±6.1) years, including 12 males. Among them, 10 patients had external commissural prolapse, and 9 patients had internal commissural prolapse. Preoperatively, all patients exhibited severe mitral regurgitation (4+), with an effective regurgitant orifice area of (0.55±0.17) cm2, left atrial volume of (104.77±36.57) ml, left ventricular end-diastolic volume of (102.29±32.47) ml, left ventricular end-diastolic dimension of (5.34±0.59) mm, and prolapse width of (1.18±0.34) cm. All procedures utilized short clips (NTR or NTW clips) to target the prolapsed commissural region and were completed successfully without intraoperative complications. At 1-month follow-up, no mortality, stroke, single-leaflet device attachment, myocardial infarction, or unplanned mitral reintervention occurred. Mitral regurgitation severity improved to ≤2+ in all patients, with left atrial volume of (74.49±33.83) ml, left ventricular end-diastolic volume of (85.90±18.05) ml, and left ventricular end-diastolic dimension of (4.93±0.37) mm (all P<0.05). Conclusion:The short-clip strategy, focusing on precise clip placement at the commissural interface, is feasible and effective for TEER in patients with severe mitral regurgitation due to commissural prolapse.

In recent decades,the incidence of human papillomavirus(HPV)-associated oropharyngeal squamous cell carcinoma(OPSCC)has shown a marked increase.Significant changes have also occurred in the OPSCC diagnosis and treatment paradigm.Deter-mining HPV status prior to treatment is now essential,and radiotherapy/chemotherapy,immunotherapy,and minimally invasive surgical techniques have progressively emerged as key modalities for managing OPSCC.However,alongside these paradigm shifts,a comprehen-sive technical consensus guiding the entire diagnostic and therapeutic process for OPSCC patients is currently lacking.Given China's large population base and the rising incidence of OPSCC,an expert panel convened to develop a clinical technical consensus on OPSCC diagno-sis and management tailored to China's specific context.This consensus aims to further enhance and standardize understanding of OPSCC management techniques among relevant healthcare professionals.

Objective:This study aimed to propose an innovative modular surgical technique and explore its safety and application value in laparoscopic total gastrectomy combined with spleen-preserving hilar lymphadenectomy for advanced proximal gastric cancer invading the greater curvature.Methods:A retrospective collection was conducted on 34 patients with proximal gastric cancer invading the greater curvature who underwent laparoscopic total gastrectomy combined with spleen-preserving hilar lymphadenectomy in the same center from October 2020 to December 2022. The technical key points, precautions and crucial steps of the modular surgical technique were summarized, and the Clinical indicators were analyzed.Results:All 34 patients successfully completed the operation under laparoscopy without conversion to open surgery. The average operation duration was 151.9±4.1 minutes, and the duration of splenic hilar lymphadenectomy was 12.9±1.5 minutes. The median intraoperative blood loss was 50(20, 50) ml, and the blood loss during splenic hilar lymphadenectomy was 5 (2, 5) ml. The median number of harvested lymph nodes was 32.0 (23.5,39.5), and the number of submitted No.10 lymph nodes was 3 (2, 4). The metastasis rate of No.10 lymph nodes was 20.6% (7/34). No patient had intraoperative complications. During the postoperative hospital stay, one patient had incision infection (Clavien-Dindo I), and one patient had pulmonary infection (Clavien-Dindo II). The time for the first postoperative feeding was 3 (2, 5) days, the time for the first postoperative flatus was 2 (2,3) days, the time for the first postoperative defecation was 3 (3, 4) days, the total postoperative drainage volume was 1047.5 (607.5,1397.5) mL, the time for postoperative drainage tube removal was 7 (6, 9) days, and the length of postoperative hospital stay was 7.0 (6.0, 9.5) days.Conclusions:The application of the innovative modular surgical technique in laparoscopic total gastrectomy combined with spleen-preserving hilar lymphadenectomy can simplify surgical process and enable safe, precise and comprehensive dissection of splenic hilar lymph nodes.

To prepare monoclonal antibody to the N protein of porcine reproductive and respiratory syndrome virus(PRRSV),BALB/c mice were immunized with the purified N protein of the PRRSV-2 strain expressed by prokaryotic expression system.Mouse splenocytes were fused with myeloma cells using hybridoma technique,hybridoma cells were identified by indirect ELISA method and indirect immunofluorescence assay(IFA),and positive hybridoma cells were screened for subclones using the limited dilution method.The results showed that a monoclonal antibody cell line 4A7 was successfully obtained,and the results of Western blot and IFA indicated that the monoclonal antibody could accurately recognize the N proteins of type 1 and type 2 PRRSV.Mean-while,the N protein gene was truncated and expressed by prokaryotic expression system,and the amino acid sequence of the B-cell antigenic epitope recognized by 4A7 was screened as 51EKPHF55 using Western blot.Comparison of epitope amino acids in the N protein gene sequences of different strains revealed that the antigenic epitope 51EKPHF55 recognized by the monoclonal antibody 4A7 has no amino acid difference in the sequences of three subtypes among the PRRSV-1 strains and nine lineages among the PRRSV-2 strains,indicating a high degree of conservation.The results of the study provide a foundation the development of PRRSV diagnostic kits and novel vaccines.

In recent decades,the incidence of human papillomavirus(HPV)-associated oropharyngeal squamous cell carcinoma(OPSCC)has shown a marked increase.Significant changes have also occurred in the OPSCC diagnosis and treatment paradigm.Deter-mining HPV status prior to treatment is now essential,and radiotherapy/chemotherapy,immunotherapy,and minimally invasive surgical techniques have progressively emerged as key modalities for managing OPSCC.However,alongside these paradigm shifts,a comprehen-sive technical consensus guiding the entire diagnostic and therapeutic process for OPSCC patients is currently lacking.Given China's large population base and the rising incidence of OPSCC,an expert panel convened to develop a clinical technical consensus on OPSCC diagno-sis and management tailored to China's specific context.This consensus aims to further enhance and standardize understanding of OPSCC management techniques among relevant healthcare professionals.

Objective:This study aimed to propose an innovative modular surgical technique and explore its safety and application value in laparoscopic total gastrectomy combined with spleen-preserving hilar lymphadenectomy for advanced proximal gastric cancer invading the greater curvature.Methods:A retrospective collection was conducted on 34 patients with proximal gastric cancer invading the greater curvature who underwent laparoscopic total gastrectomy combined with spleen-preserving hilar lymphadenectomy in the same center from October 2020 to December 2022. The technical key points, precautions and crucial steps of the modular surgical technique were summarized, and the Clinical indicators were analyzed.Results:All 34 patients successfully completed the operation under laparoscopy without conversion to open surgery. The average operation duration was 151.9±4.1 minutes, and the duration of splenic hilar lymphadenectomy was 12.9±1.5 minutes. The median intraoperative blood loss was 50(20, 50) ml, and the blood loss during splenic hilar lymphadenectomy was 5 (2, 5) ml. The median number of harvested lymph nodes was 32.0 (23.5,39.5), and the number of submitted No.10 lymph nodes was 3 (2, 4). The metastasis rate of No.10 lymph nodes was 20.6% (7/34). No patient had intraoperative complications. During the postoperative hospital stay, one patient had incision infection (Clavien-Dindo I), and one patient had pulmonary infection (Clavien-Dindo II). The time for the first postoperative feeding was 3 (2, 5) days, the time for the first postoperative flatus was 2 (2,3) days, the time for the first postoperative defecation was 3 (3, 4) days, the total postoperative drainage volume was 1047.5 (607.5,1397.5) mL, the time for postoperative drainage tube removal was 7 (6, 9) days, and the length of postoperative hospital stay was 7.0 (6.0, 9.5) days.Conclusions:The application of the innovative modular surgical technique in laparoscopic total gastrectomy combined with spleen-preserving hilar lymphadenectomy can simplify surgical process and enable safe, precise and comprehensive dissection of splenic hilar lymph nodes.

Objective:To investigate the feasibility of transcatheter edge-to-edge repair (TEER) using a short-clip strategy for patients with moderate-to-severe or greater degenerative mitral regurgitation caused by commissural prolapse.Methods:This retrospective study included patients with severe mitral regurgitation secondary to commissural prolapse who underwent TEER at the Second Affiliated Hospital of Zhejiang University School of Medicine between September 2022 and July 2024. Preoperative clinical and imaging data, intraoperative details, procedural outcomes, and 1-month postoperative follow-up results were collected.Results:A total of 19 patients were enrolled, aged (74.1±6.1) years, including 12 males. Among them, 10 patients had external commissural prolapse, and 9 patients had internal commissural prolapse. Preoperatively, all patients exhibited severe mitral regurgitation (4+), with an effective regurgitant orifice area of (0.55±0.17) cm2, left atrial volume of (104.77±36.57) ml, left ventricular end-diastolic volume of (102.29±32.47) ml, left ventricular end-diastolic dimension of (5.34±0.59) mm, and prolapse width of (1.18±0.34) cm. All procedures utilized short clips (NTR or NTW clips) to target the prolapsed commissural region and were completed successfully without intraoperative complications. At 1-month follow-up, no mortality, stroke, single-leaflet device attachment, myocardial infarction, or unplanned mitral reintervention occurred. Mitral regurgitation severity improved to ≤2+ in all patients, with left atrial volume of (74.49±33.83) ml, left ventricular end-diastolic volume of (85.90±18.05) ml, and left ventricular end-diastolic dimension of (4.93±0.37) mm (all P<0.05). Conclusion:The short-clip strategy, focusing on precise clip placement at the commissural interface, is feasible and effective for TEER in patients with severe mitral regurgitation due to commissural prolapse.