Objective To prepare specific antibodies against VP8 proteins of rotavirus(RV) genotypes P[8], P[6], and P[4], and to initially establish and verify an ELISA method for the detection of recombinant RV VP8 protein antigens.Methods Female BALB/c mice were immunized with recombinant P[8], P[6], and P[4]type VP8 proteins at a dose of60 μg per mouse with three mice for each type, and booster immunizations were administered via intramuscular injection in the leg at the same dose for 4 times at intervals of 2 to 4 days. Splenocytes from the immunized mice were fused with SP2/0 cells to screen for monoclonal antibodies(McAbs) against P[8], P[6], and P[4]type VP8 proteins. Polyclonal antibodies were obtained by immunizing rabbits. A double-antibody sandwich ELISA method for detecting recombinant RV VP8 protein antigens was initially established by optimizing the pairing combinations of capture and detection antibodies as well as their working concentration. The linearity and range of the standard curve were determined, and the precision, accuracy, and specificity of the method were verified. The established method was applied to detect intermediate samples(bacterial cell lysate, crude extract, purified solutionⅠ, and purified solutionⅡ) from the preparation and purification processes of three batches of P[8], P[6], and P[4]type VP8 proteins.Results One hybridoma cell line secreting anti-P[8]McAb was screened and named P[8]-19E7; two anti-P[6]McAb-secreting cell lines were obtained, designated as P[6]-4B8 and P[6]-8F11; and one anti-P[4]McAb-secreting cell line was identified, named P[4]-11E3. Additionally, rabbit polyclonal antibodies against P[8], P[6]and P[4]were prepared. For the P[6]antigen detection, both the capture antibody(30 μg/mL)and the detection antibody(1. 0 μg/mL) were mouse McAbs, with a standard curve range of 10. 00 to 640. 00 ng/mL. For the P[8]and P[4]antigen detection, the capture antibodies were McAbs, with the concentration of 5 μg/mL and 2. 5 μg/mL respectively, the detection antibodies were rabbit polyclonal antibodies, both at the concentration of 1. 5 μg/mL, and their standard curve ranges were 0. 63-20. 00 ng/mL and 0. 31-10. 00 ng/mL respectively. The coefficients of variation(CVs) of the six test results for the antigen content of P[8], P[6]and P[4]proteins were all less than 10%, the recovery rates ranged from 80% to 120%, and the specificity was good. The CVs of antigen recovery yield for the in-process products from three batches of purification processes were all less than 30%.Conclusion The screened monoclonal cell lines secrete antibodies that specifically target the P[8], P[6]and P[4]subtypes of VP8 protein without cross-reactivity, and exhibit high antibody titers. The recombinant RV VP8 protein antigen detection method established based on these antibodies demonstrates strong specificity, high sensitivity, and excellent precision.

AIM:To investigate the mechanism of reduced tear secretion in depression-related dry eye rats based on 5-HT/TRPV4/AQP5.METHODS:Healthy SD male rats were established with chronic unpredictable mild stress(CUMS)method to establish a depression-induced dry eye model(n=8), and the control group was blank rats(n=8). The ELISA method was used to compare the 5-hydroxytryptamine(5-HT)in serum and hippocampal tissue of the two groups, and the HE sections of lacrimal gland and AQP5 immunohistochemistry were observed. Western blot and RT-PCR were used to detect the expression of 5-HT3R, TRPV4 and AQP5 in the lacrimal gland tissue of the two groups of rats.RESULTS:The tear secretion in the depression-induced group was significantly reduced(P=0.001), the serum and hippocampal 5-HT levels were significantly reduced(all P<0.05), the expression of AQP5 antibody in the lacrimal gland immunohistochemistry was significantly lower than that in the control group(P<0.001), the expression of 5-HT3R, TRPV4 and AQP5 in the lacrimal gland was significantly reduced(all P<0.05), and no obvious inflammatory cells were found in the lacrimal gland tissue sections.CONCLUSION:Depression-related dry eye may occur through a non-inflammatory 5-HT/TRPV4/AQP5 mechanism.

AIM:To investigate the mechanism of reduced tear secretion in depression-related dry eye rats based on 5-HT/TRPV4/AQP5.METHODS:Healthy SD male rats were established with chronic unpredictable mild stress(CUMS)method to establish a depression-induced dry eye model(n=8), and the control group was blank rats(n=8). The ELISA method was used to compare the 5-hydroxytryptamine(5-HT)in serum and hippocampal tissue of the two groups, and the HE sections of lacrimal gland and AQP5 immunohistochemistry were observed. Western blot and RT-PCR were used to detect the expression of 5-HT3R, TRPV4 and AQP5 in the lacrimal gland tissue of the two groups of rats.RESULTS:The tear secretion in the depression-induced group was significantly reduced(P=0.001), the serum and hippocampal 5-HT levels were significantly reduced(all P<0.05), the expression of AQP5 antibody in the lacrimal gland immunohistochemistry was significantly lower than that in the control group(P<0.001), the expression of 5-HT3R, TRPV4 and AQP5 in the lacrimal gland was significantly reduced(all P<0.05), and no obvious inflammatory cells were found in the lacrimal gland tissue sections.CONCLUSION:Depression-related dry eye may occur through a non-inflammatory 5-HT/TRPV4/AQP5 mechanism.

Objective: To investigate the prevalence and influencing factors of menopausal syndrome among postmenopausal women in Pan'an County, Zhejiang Province, so as to provide the basis for guiding the health management of postmenopausal women. Methods: From May 2023 to April 2024, the postmenopausal women aged 40 to 69 years in Pan'an County were selected using the random cluster sampling method. Demographic information, lifestyle and prevalence of gynecological diseases were collected through questionnaire surveys. The prevalence of menopausal syndrome was assessed by modified Kupperman Score Scale. Factors affecting menopausal syndrome were analyzed by a multivariable logistic regression model. Results: A total of 816 postmenopausal women were surveyed, with an mean age of (57.63±2.92) years and a mean natural menopause age of (49.85±2.13) years. There were 574 cases with menopausal syndrome, with a prevalence of 70.34%. Flashes and sweating, insomnia and irritability were common symptoms, accounting for 62.87%, 47.43% and 41.18%, respectively. Multivariable logistic regression analysis showed that monthly personal income of ≤5 000 yuan (<3 000 yuan, OR=3.124, 95%CI: 1.829-5.335; 3 000-5 000 yuan, OR=2.399, 95%CI: 1.370-4.201) and having gynecological diseases (OR=1.970, 95%CI: 1.292-3.004) were associated with a higher risk of menopausal syndrome, while average (OR=0.141, 95%CI: 0.072-0.276) or sufficient sleep quality (OR=0.095, 95%CI: 0.049-0.185) were associated with a lower risk of menopausal syndrome. Conclusion The prevalence of menopausal syndrome among postmenopausal women in Pan'an County is relatively high, and is mainly influenced by personal economic status, sleep quality and the presence of gynecological diseases.

[Objective] To obtain accurate data on alanine aminotransferase (ALT) levels among blood donors in China and to explore the necessity of adjusting the qualification criteria for ALT. [Methods] A collaborative study was conducted involving 26 blood centers and 7 central blood stations with an annual testing volume exceeding 100 000 samples. Between December 1 and 15, 2024, pre-donation ALT testing was suspended for 1-2 days for all whole blood donations. ALT levels were measured only post-donation using standard laboratory equipment and reagents. All transfusion-transmitted infectious disease-related serological and nucleic acid testing, including hepatitis E virus (HEV) RNA testing, were performed. Within one week of testing completion, anonymized data on basic donor information, routine test results, and HEV RNA results were collected and statistically analyzed. [Results] A total of 21 345 blood donors were included in the study, with an ALT disqualification rate of 7.6% (1 623/21 345). The disqualification rate was 9.6% (1 453/15 205) for males and 2.8% (170/6 140) for females. There were significant regional variations in both the disqualification rates and levels of ALT, with Shaanxi Province exhibiting the highest disqualification rate (12.3%, 87/710) and Yunnan Province the lowest (2.9%, 19/652). Among the provinces (autonomous regions and municipalities), Beijing recorded the lowest ALT levels. ALT levels varied across different age groups and genders. Among all samples tested by HEV RNA, the HEV RNA positive rate was 0.29‰ (6/21 003). HCV infection was found to directly affect ALT levels, while HBV, HIV, syphilis, and HEV infections did not significantly impact ALT disqualification rates. It is recommended to adjust the ALT qualification criteria to twice the upper limit of the clinical reference range, which would increase the number of eligible blood donations by 6.61% (1 293/19 550). [Conclusion] In China, the ALT levels of blood donors are correlated with gender, age, geographical region, and HCV infection status. Appropriately raising the ALT eligibility criteria to ≤100 U/L for male donors and ≤80 U/L for female donors could expand the pool of eligible donors and reduce the blood discard rate while ensuring blood safety.

Objective: To explore the relationship between school sports environment and physical activity levels of middle school students, so as to provide theoretical and empirical support for optimizing school sports environment and enhance adolescent physical activity. Methods: Using multi-stage random cluster sampling, from September to December 2023, 1 329 junior and senior high school students from Xuancheng City of Anhui Province, Lianyungang City of Jiangsu Province, Wuhan City of Hubei Province, Qiqihar City and Suihua City of Heilongjiang Province, and Shenzhen City of Guangdong Province were selected. The International Physical Activity Questionnaire-Short Form (IPAQ-SF) assessed students physical activity levels, and the questionnaire on the characteristics of school sports environment was developed to evaluate the factors of school sports environment. Multivariate ordered Logistic regression was performed to analyze the correlation between school sports environment factors and physical activity levels, and the analytic hierarchy process determined the weight of key influencing factors. Results: The results showed that weekly vigorous physical activity time was [60 (25, 90)] minutes, moderate physical activity time was [60 (30, 90)] minutes, light physical activity time was [105 (40, 200)] minutes, and sedentary behavior time was [ 3 300 (2 100, 4 500)] minutes, only 10.53% of the students met World Health Organization physical activity recommendations, and 89.69% of the students averaged >8 h daily sedentary time. Multivariate ordered Logistic regression showed that adequate sports equipment significantly promoted physical activity across all intensities and reduced sedentary time ( OR = 4.97, 11.54, 4.03, 0.11); diverse sports activities improved vigorous and moderate physical activity while reducing sedentary time ( OR =4.20, 14.06, 0.17); and peer encouragement was associated with increased low-intensity physical activities and decreased sedentary time ( OR =10.40, 0.15)( P <0.05). The analytic hierarchy process weighting analysis identified the top three influential factors related to physical activity among middle school students: sufficient sports equipment, varied physical education activities, frequent peer encouragement, the influence weight accounts for 23.55% , 14.18% and 11.77% of the total, respectively. Conclusion Key school sports environmental factors for adolescent physical activity level include ensuring adequate sports equipment and class availability, diversifying activity content, fostering peer support, and cultivating an active sports culture and a comprehensive approach encourage students participation in extracurricular physical activities.

INTRODUCTION: Trastuzumab deruxtecan (T-DXd) has revolutionised treatment for metastatic breast cancer (MBC). While effective, its high cost and toxicities, such as fatigue and nausea, pose challenges. METHOD: Medical records from the Joint Breast Cancer Registry in Singapore were used to study MBC patients treated with T-DXd (February 2021-June 2024). This study was conducted to address whether reducing dose intensity and density may have an adverse effect on treatment outcomes. RESULTS: Eighty-seven MBC patients were treated with T-DXd, with a median age of 59 years. At the time of data cutoff, 32.1% of patients were still receiving T-DXd. Over half (54%) of the patients received treatment with an initial relative dose intensity (RDI) of <;85%. Overall median real-world progression-free survival (rwPFS) was 8.1 months. rwPFS was similar between RDI groups (<85%: 8.7 months, <85%: 8.1 months, P=0.62). However, human epidermal growth receptor 2 (HER2)-positive patients showed significantly better rwPFS outcomes compared to HER2-low patients (8.8 versus 2.5 months, P<0.001). Only 16% with central nervous system (CNS) involvement had CNS progressive disease on treatment. No significant progression-free survival (PFS) differences were found between patients with or without CNS disease, regardless of RDI groups. Five patients (5.7%) developed interstitial lung disease (ILD), with 3 (3.4%) having grade 3 events. Two required high-dose steroids and none were rechallenged after ILD. There were no fatalities. CONCLUSION Our study demonstrated that reduced dose intensity and density had no significant impact on rwPFS or treatment-related toxicities. Furthermore, only 5.7% of patients developed ILD. T-Dxd provided good control of CNS disease, with 82% of patients achieving CNS disease control.
Humans ; Female ; Breast Neoplasms/mortality* ; Middle Aged ; Trastuzumab/adverse effects* ; Aged ; Adult ; Singapore/epidemiology* ; Antineoplastic Agents, Immunological/adverse effects* ; Camptothecin/adverse effects* ; Immunoconjugates/adverse effects* ; Retrospective Studies ; Progression-Free Survival ; Receptor, ErbB-2/metabolism* ; Neoplasm Metastasis ; Dose-Response Relationship, Drug ; Treatment Outcome ; Registries

BACKGROUND: Oral squamous cell carcinoma (OSCC) is a prevalent type of cancer with a high mortality rate in its late stages. One of the major challenges in OSCC treatment is the resistance to epidermal growth factor receptor (EGFR) inhibitors. Therefore, it is imperative to elucidate the mechanism underlying drug resistance and develop appropriate precision therapy strategies to enhance clinical efficacy. METHODS: To evaluate the efficacy of the combination of the Ca 2+ /calmodulin-dependent protein kinase II (CAMK2) inhibitor KN93 and EGFR inhibitors, we performed in vitro and in vivo experiments using two FAT atypical cadherin 1 ( FAT1 )-deficient (SCC9 and SCC25) and two FAT1 wild-type (SCC47 and HN12) OSCC cell lines. We assessed the effects of EGFR inhibitors (afatinib or cetuximab), KN93, or their combination on the malignant phenotype of OSCC in vivo and in vitro . The alterations in protein expression levels of members of the EGFR signaling pathway and SRY-box transcription factor 2 (SOX2) were analyzed. Changes in the yes-associated protein 1 (YAP1) protein were characterized. Moreover, we analyzed mitochondrial dysfunction. Besides, the effects of combination therapy on mitochondrial dynamics were also evaluated. RESULTS: OSCC with FAT1 mutations exhibited resistance to EGFR inhibitors treatment. The combination of KN93 and EGFR inhibitors significantly inhibited the proliferation, survival, and migration of FAT1 -mutated OSCC cells and suppressed tumor growth in vivo . Mechanistically, combination therapy enhanced the therapeutic sensitivity of FAT1 -mutated OSCC cells to EGFR inhibitors by modulating the EGFR pathway and downregulated tumor stemness-related proteins. Furthermore, combination therapy induced reactive oxygen species (ROS)-mediated mitochondrial dysfunction and disrupted mitochondrial dynamics, ultimately resulting in tumor suppression. CONCLUSION Combination therapy with EGFR inhibitors and KN93 could be a novel precision therapeutic strategy and a potential clinical solution for EGFR-resistant OSCC patients with FAT1 mutations.
Humans ; ErbB Receptors/metabolism* ; Mouth Neoplasms/metabolism* ; Cell Line, Tumor ; Animals ; Drug Resistance, Neoplasm/genetics* ; Cadherins/metabolism* ; Carcinoma, Squamous Cell/metabolism* ; Mice ; Mutation/genetics* ; Mice, Nude ; Protein Kinase Inhibitors/therapeutic use* ; Cetuximab/pharmacology* ; Afatinib/therapeutic use* ; Cell Proliferation/drug effects* ; Signal Transduction/drug effects*