Objective: To develop a prediction model for mild cognitive impairment (MCI) risk among the elderly, so as to provide a tool for MCI early screening. Methods : From July 2022 to September 2024, a multi-stage stratified random cluster sampling method was used to recruit permanent residents aged ≥65 years from the Xinjiang Uygur Autonomous Region as study participants. Data on sociodemographic characteristics, nutritional status, body composition indices, bone mineral density, and handgrip strength were collected through questionnaires and physical examinations. Sarcopenia was defined based on appendicular skeletal muscle index and handgrip strength. MCI was assessed using the Mini-Mental State Examination, with adjustments for educational level. Participants were randomly divided into a training set and a validation set in a 7∶3 ratio. LASSO regression and multivariable logistic regression models were employed to screen for predictors and construct an MCI risk prediction model. The predictive performance of the model was evaluated using receiver operating characteristic (ROC) curve and decision curve analysis (DCA). Results: A total of 1 641 participants were surveyed, including 755 males (46.01%) and 886 females (53.99%). The majority of participants were aged 65-<75 years, comprising 1 154 individuals (70.32%). MCI was detected in 517 participants, corresponding to a detection rate of 31.51%. Resultsfrom LASSO regression and multivariate logistic regression analysis showed that residence (rural, OR = 2.323, 95% CI: 1.682-3.210), age (75-<85 years, OR = 1.405, 95% CI: 1.019-1.937; ≥85 years, OR = 3.655, 95% CI: 1.696-7.875), educational level (primary school, OR = 0.341, 95% CI: 0.247-0.472; junior high school, OR = 0.255, 95% CI: 0.160-0.408; high school, OR = 0.286, 95% CI: 0.154-0.531; bachelor's degree or above, OR = 0.120, 95% CI: 0.041-0.351), history of alcohol consumption (yes, OR = 3.216, 95% CI: 2.164-4.779), risk of malnutrition (yes, OR = 1.464, 95% CI: 1.064-2.014), sarcopenia (yes, OR = 3.197, 95% CI: 2.332-4.385), and waist-to-hip ratio (abnormal, OR = 1.540, 95% CI: 1.159-2.048) were identified as predictive factors for MCI among the elderly. In the training set, the area under the ROC curve, sensitivity, and specificity were 0.788, 0.719, and 0.712, respectively. In the validation set, the corresponding values were 0.784, 0.913, and 0.542, respectively. DCA demonstrated that the model provided a higher clinical net benefit for predicting MCI risk when the risk threshold probability ranged from 0.124 to 0.764. Conclusion The prediction model developed in this study demonstrates good discriminative ability and clinical utility, indicating its substantial value for predicting the MCI risk among the elderly.

Driven by the growing practical need to accelerate drug development and the continuous innovation of trial design in recent years， the number of protocol amendments during clinical trials have gradually increased， and the changed contents have become more flexible and complex， which significantly heightens the difficulty of ethical review on amendments. Against this backdrop， it is of great importance to fully leverage the role and responsibilities of ethics committees， effectively control clinical trial risks， and ensure subject safety. This paper analyzed development trends of protocol amendments in recent years， sorted out requirements for protocol amendments in Chinese regulations and guiding principles， and examined difficulties of amendment ethical review in practical work. Based on these， targeted strategies and recommendations were proposed， namely， strengthening the integration with scientific review， enhancing the formal review， adjusting the scope of review according to approval notifications， and adopting appropriate review methods， with a view to providing insights and references for the management of the amendment ethical review in drug clinical trials.

Objective: To explore the association between the HLA antibody positivity rate in female blood donors and pregnancy history, number of pregnancies, interval from the last pregnancy to blood donation, and age, to identify associated variables using a univariate generalized additive model (GAM), and to further analyze the predictive role of characteristic variables for HLA antibody positivity using a random forest model. Methods: HLA antibody detection was performed on 391 female blood donors using the Luminex immunomagnetic bead method. The correlation between pregnancy-related factors and HLA antibodies was analyzed using the Chi-square test. Based on R software, a univariate GAM was first constructed to analyze the association types between characteristic variables and the HLA antibody positivity rate, followed by the construction of a random forest model to evaluate the predictive value of the variables. Results: Among the 391 female blood donors without a transfusion history, the overall HLA antibody positivity rate was 26.34%. The positivity rate in donors with a pregnancy history was significantly higher than that in those without (30.09% vs 9.72%, P<0.05), and HLA antibody positivity rate increased linearly with the number of pregnancies (P<0.05). In the univariate GAM, age and number of deliveries exhibited a non-linear association with the HLA antibody positivity rate (the positivity rate increased sharply between 25-35 years of age and stabilized after 3 deliveries). Besides, the interval from the last pregnancy to blood donation showed a linear association with the HLA antibody positivity rate, and the positivity rate decreased as the interval prolonged (P<0.05). In the random forest model, age (mean decrease gini=29.26) and interval from the last pregnancy to blood donation (mean decrease gini=22.02) were core predictive variables: age was more conducive to identifying positive samples, while the interval from the last pregnancy to blood donation was more helpful for excluding negative samples. The number of deliveries (mean decrease accuracy=16.98) made a significant contribution to predicting positive samples, whereas the number of abortions had no impact. The model had an AUC of 0.583 (95% CI: 0.593 8-0.770 2), indicating a certain predictive value. Conclusion: The associated variables identified by the univariate GAM model, including age, interval from the last pregnancy to blood donation, and number of deliveries, provide a basis for key variables in the random forest model. All three variables have predictive value for HLA antibody positivity, which can provide evidence-based support for personalized transfusion management and stratified screening of female blood donors in this region.

ObjectiveTo systematically assess the public health governance capacity in Zhejiang Province, to conduct an in-depth analysis of its strengths and weaknesses, so as to provide scientific basis and strategic recommendations for further enhancement. MethodsA systematic collection of policy documents, public information reports, and research literature related to public health governance capacity in Zhejiang Province from 2002 to 2023 was conducted (encompassing a total of 1 263 policy documents, 138 pieces of information reports and 631 research articles). Based on the evaluation criteria suitable for public health systems previously developed by the research team, the basic status and magnitude of change in public health governance capacity in Zhejiang Province was evaluated. Additionally, normative gap analyses were employed to identify the strengths and weaknesses. ResultsZhejiang Province ranked 4th nationwide in terms of public health governance capacity with a score of 733.4 points (1 000.0-point maximum). The province has effectively implemented the principle of health first (scoring 698.5 points in the assessment of health-first strategy implementation) and attached sufficient importance to health-related goals (scoring 658.2 points in the scientific rationality of goal setting). However, the implementation of inter-departmental coordination and incentive mechanisms only scored 178.7 points, the feasibility of management and monitoring mechanisms scored even lower at only 144.0 points, and the coverage of incentive mechanisms scored 286.0 points. ConclusionZhejiang Province has effectively implemented its health first strategy and attached great importance to health targets, but still needs to strengthen cross-departmental coordination mechanisms and health-oriented incentives.

Purpose To investigate the presence,proportion,clinical significance and origin of tumor cells with a macrophage phenotype in tumor tissues of patients with diffuse large B-cell lymphoma(DLBCL),and to explore wheth-er CD68 positive tumor cells can be induced in DLBCL cell lines in vitro.Methods The presence of CD68+CD163+CD20+PAX5-cells in the samples of DLBCL patients was first qualitatively detected by multiplex immunofluorescence staining,and then the proportion of CD79a+B lymphocytes,CD68+macrophages,and CD68+CD79a+double-positive cells were quantified.Patients were grouped according to the proportion of double-positive cells,and the differences in prognosis and clinicopathological features of DLBCL patients between subgroups were investigated.For cases with posi-tive BCL6 gene locus breaks,co-localization of CD68 with BCL6 gene breakapart was performed using combined immu-nofluorescence and immunological in situ hybridization to ascertain the tumor nature of B cell with a macrophage pheno-type.DLBCL cell lines(OCI-LY3,SU-DHL2)were treated with phorbol myristate acetate(PMA),and changes in the proteins levels of CD68 and PAX5 proteins were detected by flow cytometry.Quantitative real-time PCR(qRT-PCR)was used to detect mRNA levels of PAX5,an important transcription factor for B cell differentiation and develop-ment,and macrophage-related genes(CD68,ARG1,CD163,CD206,Dectin-1,PU.1,C/EBPα,C/EBPβ).In ad-dition,PMA-treated DLBCL cell lines(OCI-LY3,SU-DHL2)were co-incubated with pH-sensitive fluorescent dye pHrodo to detect the phagocytosis ability of PMA-treated DLBCL cells.Results The percentage of CD68+B lympho-cytes in 50 patients with DLBCL varied from 0 to 9.3％,and the overall survival(OS)ranged from 0.008 2 to 4.2 years.Patients with the low CD68+B lymphocytes group exhibited a significantly lower OS compared to those in the high CD68+B lymphocytes group(P=0.039).There was a significant difference in the molecular typing of DLBCL patients(P=0.009 5)between different subgroups for the proportion of CD68+B lymphocytes.CD68+B lymphocytes were derived from tumor cells in DLBCL patients.The proportion of CD68+cells and CD68+PAX5-cells significantly increased in DLBCL after treatment with PMA(P＜0.05).The other macrophage markers CD68,ARG1,CD 163,CD206,Dectin-1,PU.1,C/EBPα,C/EBPβ,and the important B-cell transcription factor PAX5 were significantly different from the control group in terms of relative mRNA expression(P＜0.05).Cellular phagocytosis was enhanced after PMA treatment of DLBCL cells.Conclusion Diffuse large B-cell lymphoma tumor tissue contains a certain per-centage of CD68+neoplastic B lymphocytes.The proportion of CD68+B lymphocytes is correlated with patient progno-sis and molecular typing.DLBCL cell lines can be induced to differentiate into CD68+tumor cells in vitro.

Objective To analyze the differences and correlations of imaging parameters in patients with Parkinson's disease(PD)combined with cerebral small vessel disease(CSVD)with different cognitive functions based on MRI.Methods A retrospective selection was conducted on 192 PD patients combined with CSVD.According to the Montreal cognitive assessment(MoCA)and mini-mental state examination(MMSE)scores,they were divided into cognitive impairment PD group(n=69)and cognitive normal PD group(n=123),and the clinical data and MRI imaging parameters of the two groups were compared.Results The age and drinking history of the cog-nitive impairment PD group were higher than those of the cognitive normal PD group(P<0.05);the lacune of presumed vascular origin(LPVO)score,periventricular white matter hyperintensity(PVWMH)score,basal ganglia-perivascular space(BG-PVS)score,brain atrophy(BA)score,and CSVD total score in the cognitive impairment PD group were higher than those in the cognitive normal PD group(P<0.05),while there was no statistically significant difference in deep white matter hyperintensity(DWMH)score,cen-trum semiovale perivascular space(CS-PVS)score,and cerebral microbleed(CMB)score between the two groups(P>0.05);Multi-variate logistic analysis showed that age,LPVO score,BG-PVS score,BA score,and CSVD total score were independent influencing factors of cognitive impairment(P<0.05).Conclusion There are differences in imaging parameters such as LPVO score,PVWMH score,BG-PVS score,BA score,and CSVD total score among PD patients combined with CSVD with different cognitive functions,and they are correlated with MMSE and MoCA scores.

Objective:To preliminarily explore whether sirtuin1 (SIRT1) activation can inhibit neuronal ferroptosis in rats after traumatic brain injury (TBI) by regulating hypoxia-inducible factor-1α (HIF-1α)-mediated glycolysis.Methods:(1) Six SD rats were randomly divided into sham-operated group and TBI group, with 3 rats in each group; TBI model in the TBI group was established by hydraulic impact method, and rats in the sham-operated group underwent same surgery without impact. Cortical tissues of the two groups were sent for tandem mass tag (TMT) labeled quantitative proteomics detection to analyze the differential expression proteome; Kyoto encyclopedia of genes and genomes (KEGG) and gene set enrichment analysis (GSEA) were used to detect pathway enrichment of the screened differential proteins. (2) Twelve SD rats were randomly divided into sham-operated group and 1-day, 3-day and 7-day post-TBI groups, with 3 rats in each group. Treatment methods were the same as above; Western blotting was used to detect SIRT1 protein expression. (3) Forty-eight rats were randomly divided into sham-operated group, TBI group, TBI+vehicle group and TBI+SIRT1 agonist group, with 12 rats in each group; rats in the sham-operated group and TBI group accepted treatment as above; rats in the TBI+SIRT1 agonist group were intraperitoneally injected with SRT1720 (dissolved in ≤ 5% dimethyl sulfoxide, at a dose of 20 mg/kg) within 30 minutes after modeling, twice a day (with an interval of 12 hours); and rats in the TBI+vehicle group were injected with same dose of dimethyl sulfoxide at the same time. One d after modeling, neurological deficit was assessed using modified Neurological severity score (mNSS), brain water content was measured by dry-wet weight method, histopathological changes in the cortical lesions were observed by HE staining, mitochondrial ultrastructure was examined by transmission electron microscopy, malondialdehyde (MDA) content and superoxide dismutase (SOD) activity in the brain tissues were detected by colorimetry, and protein expressions of SIRT1, HIF-1α (key protein in the glycolytic pathway), glutathione peroxidase 4 (GPX4, key protein in the ferroptosis pathway), and acyl-CoA synthetase long-chain family member 4 (ACSL4, key protein in the ferroptosis pathway) were evaluated by Western blotting.Results:(1) KEGG analysis revealed that the glycolysis pathway and HIF-1 signaling pathway were obviously enriched in the cortical tissues of rats in the TBI group compared with the sham-operated group; GSEA showed that the HIF-1 signaling pathway (mmu04066) and ferroptosis pathway (mmu04216) gene sets in the cortical tissues of rats in the TBI group exhibited enrichment trends compared with those in the sham-operated group. (2) Compared with the sham-operated group, the 1-day, 3-day, and 7-day post-TBI groups had significantly decreased SIRT1 protein expression ( P<0.05), with the most prominent decline in 1-day post-TBI group. (3) Compared with the TBI+vehicle group, rats in the TBI+SIRT1 agonist group showed significantly reduced mNSS score and brain tissue water content (9.83±1.17 vs. 7.66±1.21; [83.62±0.91]% vs. [80.09±0.68]%, P<0.05). HE staining indicated clearer structure of the cortical area at the injury sites, and improved neuron morphology in the TBI+SIRT1 agonist group compared with those in the TBI+vehicle group; and transmission electron microscopy showed reduced mitochondrial shrinkage and partial restoration of cristae structures in the TBI+SIRT1 agonist group compared with those in the TBI+vehicle group. Compared with the TBI+vehicle group, the TBI+SIRT1 agonist group exhibited significantly decreased MDA content ([62.72±9.20] nmol/g vs. [39.34±3.48] nmol/g), increased SOD activity ([1.95±0.23] U/mg vs. [2.48±0.14] U/mg), elevated GPX4 protein expression (0.37±0.04 vs. 0.46±0.03), and decreased HIF-1α and ACSL4 protein expressions (1.16±0.15 vs. 0.81±0.12; 1.14±0.06 vs. 1.29±0.04), with significant differences ( P<0.05). Conclusion:SIRT1 activation can exert neuroprotective effect by inhibiting HIF-1α-mediated glycolysis and reducing neuronal ferroptosis after TBI.

AIM:This study aims to explore the mechanisms of influenza A virus(IAV)-induced macrophage inflammatory injury based on the interleukin-6(IL-6)/signal transducer and activator of transcription 3(STAT3)signaling loop and investigate the intervention effects of Ma-Xing-Shi-Gan decoction(MXSGD)-medicated serum.METHODS:RAW264.7 and BV2 cells were divided into control,Janus kinase(JAK)/STAT signaling pathway activator,inhibitor,model,oseltamivir,antiviral particle,and MXSGD groups.After IAV modeling and serum interventions,the cells were cultured for 24 and 48 h,and the indicators were detected and analyzed.ELISA,RT-qPCR,Western blot,and immuno-fluorescence assay were used to detect the secretion levels of IL-6 in the cell culture supernatant,IL-6 and STAT3 mRNA expressions,protein expression of STAT3,and expression levels of phosphorylated STAT3(p-STAT3),respectively.Pearson correlation analysis was used to evaluate the correlation between p-STAT3 and IL-6 in the two cell types.A co-cul-ture model of the two cells was constructed,and the secretion levels of IL-6 in the cell culture supernatant was measured.Molecular docking analyses were performed for STAT3 and MXSGD.RESULTS:After IAV simulation,the secretion lev-els of IL-6 in the cell culture supernatant,mRNA expression levels of IL-6 and STAT3,and protein expression levels of STAT3 and p-STAT3 in both cell lines were elevated(P<0.05 or P<0.01).Pearson correlation analysis revealed that p-STAT3 expression was positively correlated with IL-6 expression.The secretion levels of IL-6 in the co-culture model in-creased(P<0.01).MXSGD down-regulated the secretion levels of IL-6 in the cell culture supernatant mRNA,expression levels of IL-6 and STAT3,and protein expression levels of STAT3 and p-STAT3 in two kinds of cells(P<0.05 or P<0.01),and inhibited the secretion levels of IL-6 in co-culture models.STAT3 demonstrated good binding energies for liquiritin,amygdalin,and ephedrine.CONCLUSION:IAV can induce inflammatory injury in macrophages,and its mechanism may be related to activation of the IL-6/STAT3 signaling loop.MXSGD may alleviate the pathogenic effects of IAV by modulating the signaling loop.

Objective:To investigate the association between high-risk human papillomavirus (hrHPV) persistent infection and vaginal microecology.Methods:A total of 53 women were enrolled in the gynecological clinic of Beijing Obstetrics and Gynecology Hospital from January 2020 to January 2021, including 7 women without HPV and 46 women with hrHPV infection. Among the hrHPV infected women, 24 woemn who did not use any drugs were classified as the observation group and the other 22 women who were given standardized interferon vaginal administration for 3 months were regarded as the treatment group. Vaginal secretions of all women were taken for Gram-stained microecological test at the time of enrollment and at the 4, 8, and 12 month follow-up. HPV turning negative was taken as the end point of follow-up.Results:(1) Women of hrHPV persistent infection in the observation and treatmnet groups had more times of abortions ( P=0.180). (2) The hrHPV negative conversion rate was 17% (4/24) in the observation group and 36% (8/22) in the treatment group, but the difference was not significant ( P=0.183). The median hrHPV negative conversion time were 11.0 months and 7.5 months in the observation and treatment groups, respectively, and the difference was statistically significant ( P=0.001). (3) Vaginal microecology was generally normal at the time of enrollment and at the end of follow-up in women with HPV natural negative conversion in the observation group. While vaginal microecological disorders were more common in women with hrHPV persistent infection in the observation and treatmnet groups, including high vaginal pH value, poor vaginal cleanliness, poor grade of Lactobacillus and increased vaginal clutter bacteria, and the vaginal microecological situation did not improve after the 12-month follow-up. (4) In the treatment group, women who turned HPV negative within six months all had normal vaginal microecology when enrollment (5/5). While those who turned negative six months later had a higher proportion of vaginal clutter bacteria (2/3), a poor grade of Lactobacillus (2/3) and a higher proportion of vaginal dysbiosis (2/3). Conclusions:(1) Interferon therapy could shorten the negative turning time of hrHPV. (2) Women with normal vaginal microecology have the ability to naturally clear hrHPV. (3) The vaginal microecological Gram-stain test has limited value in predicting hrHPV clearance, perhaps due to its inability to detect Lactobacillus subtypes.

Objective:To establish a rat model of vulvovaginal candidiasis (VVC) and to directly observe the histopathological and ultrastructural characteristics of vaginal mucosal barrier after Candida albicans infection and treatment with Lactobacillus crispatus.Methods:Female unmated SD rats were used to establish the VVC model and divided into three groups (normal group, VVC group, and Lactobacillus group; n=6 per group). Lactobacillus group received intravaginal administration of Lactobacillus crispatus suspension, while rats in VVC group and normal group were infused with phosphate buffered solution instead. Vaginal tissues were collected on day 4 post-treatment for HE staining and transmission electron microscopy (to observe ultrastructural pathological changes). Results:The results of HE staining revealed the disruption and desquamation of vaginal epithelium, necrotic epithelial tissues, neutrophil infiltration in Candida albicans-infected rats. Lactobacillus crispatus intervention restored the damaged vaginal mucosal structure (mucosal layers and thickness) to normal levels, mucosal layers of Lactobacillus group and normal group were 9.50±1.38 vs 10.67±1.03 ( P=0.226), mucosal thickness of Lactobacillus group and normal group were (116.50±12.14) vs (130.33±13.91) μm ( P=0.211). The results of transmission electron microscopy revealed intercellular desmosome rupture, loss of microvilli and glycocalyx on superficial cells, and mitochondrial swelling in Candida albicans-infected rats. Lactobacillus crispatus intervention restored the damaged vaginal mucosal ultrastructures (mitochondria and intercellular connections, etc.) to normal levels. Conclusions:Fungal infection severely disrupte the vaginal mucosal barrier in rats. Lactobacillus crispatus could restore the vaginal mucosal barrier and epithelial ultrastructures.