Objective:To investigate the cognition and learning habits of different types of postgraduates and evaluate learning effect and its potential risk factors on clinical epidemiology in a medical college, so as to provide relevant data for improving the teaching method and learning effect of clinical research methods for postgraduates.Methods:A cross-sectional study design was conducted to enroll all the postgraduates of Grade 2020 in a military medical school. A self-filled questionnaire was adopted to collect data. The discrepancy of cognition and learning habits between different types of postgraduates was evaluated by univariate analysis. Discussion was conducted to clarify the potential risk factors of learning effect. t tests or Mann-Whitney U tests were conducted to test the differences between groups for continuous variables. Chi-squared tests or McNemar tests were applied to evaluate the difference between groups for categorical variables. Results:A total of 652 postgraduate students were enrolled for analysis, including 409 master students (62.7) and 243 doctoral students (37.3). The proportion of doctoral students who have heard of clinical epidemiology ( χ2=19.99, P<0.001), who have learned clinical epidemiology ( χ2=9.20, P=0.002), who are interested in ( χ2=11.41, P=0.001) and think the course is important ( χ2=10.71, P=0.001), who previewed before class( χ2=11.21, P=0.001), reviewed after class ( χ2=3.29, P=0.001) and actively discuss in class ( χ2=11.64, P=0.001) is significantly higher than that of master students, the difference was statistically significance. The average score of all the postgraduates was (5.50±1.62) points before teaching and (7.47±1.90) points after teaching, the difference was statistically significant ( t=-23.49, P<0.001). After teaching, the grades of full-time students improved more than that of part-time graduate students, there was statistical significance in the master group ( t=4.41, P<0.001), while not in the doctor group ( t=0.94, P=0.351). Conclusions:The mastery of key points on clinical epidemiology have significantly improved after teaching among the postgraduates of different types. Different teaching methods and processes should be adopted to the variety of postgraduates according to their knowledge foundations and shortcomings. Besides, standardizing their learning habits are of certain significance to improve the learning effect.

Objective To understand the genotyping of human immunodeficiency virus (HIV) co-infected hepatitis C virus (HCV) patients in Yunnan Province, and to analyze the differences in viral load, biochemical indicators, and blood routine indicators among different genotypes, in order to provide a laboratory basis for the diagnosis and clinical treatment of HIV/HCV co-infected patients. Methods From November 2022 to June 2023, the serum samples and basic information of patients diagnosed with HIV/HCV co-infection were collected in the antiviral outpatient clinic of Yunnan Provincial Hospital of Infectious Diseases. The HCV viral load was detected by one-step qRT-PCR amplification, the positive samples were sequenced, and genotyping was determined based on NS5 gene sequence. The differences in biochemical and blood routine indexes between HIV patients co-infected with different HCV genotypes and low/high viral loads were analyzed. Results A total of 126 HIV/HCV co-infected patients were collected, including 20 HCV genotype 1 (15.9%), 91 HCV genotype 3 (72.2%), and 15 HCV genotype 6 (11.9%). The maximum and minimum viral load of the three HCV genotypes were as follows: HCV type 1 (1.0×108, 4.8×104 IU/mL), HCV type 3 (2.2×108, 2.9×102 IU/mL), and HCV type 6 (8.1×107, 6.8×104 IU/mL). The results showed that there was no significant difference between HIV co-infection with different genotypes of HCV and three HIV treatment schemes, including nucleoside reverse transcriptase inhibitors+integrase strand transfer inhibitors (NRTIs+INSTIs), nucleoside reverse transcriptase inhibitors+non-nucleoside reverse transcriptase inhibitors (NRTIs+NNRTIs) and nucleoside reverse transcriptase inhibitors+protease inhibitor (NRTIs+PLs), and the viral load of patients (P>0.05). The analysis of biochemical indexes such as total bilirubin (TBIL), direct bilirubin (DBIL), alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatinine (CREA), and blood routine indexes such as white blood cell (WBC), red blood cell (RBC), hemoglobin (HGB), platelet (PLT), mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) among different HCV genotypes and low/high viral loads showed that there was no significant difference in biochemical indexes and blood routine indexes between low/high viral loads of HIV co-infected HCV patients (P>0.05); however, the biochemical indicators TBIL, IBIL and MCHC were significantly different statistically between patients with genotype 3 HCV infection and those with genotype 1 HCV infection (P<0.05), while other biochemical and blood routine indexes were not statistically different among different HCV genotypes (P>0.05). Conclusions There are six subtypes of HCV co-infection in HIV patients in Kunming, Yunnan Province, including three genes of genotype 1, 3, and 6. Among them, genotype 3 HCV is the main prevalent genetic virus among HIV co-infected populations. The TBIL, IBIL and MCHC values of HIV patients co-infected with HCV type 3 are different from those infected with HCV type 1.

Objective:To investigate the distribution characteristics of cognition-related lifestyles of elderly in communities and explore the integrated effects on early cognitive decline.Methods:The participants were from the Project of Prevention and Intervention of Neurodegenerative Disease for Elderly in China. A total of 2 537 older adults aged ≥60 years without dementia in the 2015 baseline survey and the 2017 follow-up survey were included. The information about their cognition-related lifestyles, including physical exercise, social interaction, leisure activity, sleep quality, smoking status, and alcohol consumption, were collected through questionnaire survey and the integrated scores were calculated. Multivariate logistic regression analysis was used to assess the association between integrated cognition-related lifestyle score and early cognitive decline.Results:In the 2 537 older adults surveyed, 28.7% had score of 5-6, while only 4.8% had high scores for all 6 healthy lifestyles. Significant differences in healthy lifestyle factor distributions were observed between men and women. Multivariate logistic regression model showed that the risks for early cognitive decline in the older adults who had lifestyle score of 4 and 5-6 were lower than that in those with lifestyle score of 0-3 ( OR=0.683, 95% CI: 0.457-1.019; OR=0.623, 95% CI: 0.398-0.976; trend P=0.030). In the women, the risks for early cognitive decline was lower in groups with score of 4 and 5-6 than in group with score of 0-3 ( OR=0.491, 95% CI: 0.297-0.812; OR=0.556, 95% CI: 0.332-0.929; trend P=0.024). Conclusion:Cognition-related healthy lifestyles are associated with significantly lower risk for early cognitive decline in the elderly, especially in women.

BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most common malignant tumors in the world. The ubiquitin-specific peptidase 25 (USP25) protein has been reported to participate in the development of several cancers. However, few studies have reported its association with HCC. In this study, we aimed to investigate the function and mechanism of USP25 in the progression of HCC. METHODS: We analyzed USP25 protein expression in HCC based on The Cancer Genome Atlas (TCGA) and International Cancer Genome Consortium (ICGC) database cohorts. Then, we constructed USP25-overexpressing and USP25-knockdown HepG2, MHCC97H, and L-O2 cells. We detected the biological function of USP25 by performing a series of assays, such as Cell Counting Kit-8 (CCK-8), colony formation, transwell, and wound healing assays. Western blotting and quantitative real-time polymerase chain reaction (qRT-PCR) analyses were performed to detect the interaction between USP25 and the Wnt/β-catenin signaling pathway. The relationship between USP25 and tripartite motif-containing 21 (TRIM21) was assessed through mass spectrometry and co-immunoprecipitation (Co-IP) analysis. Finally, we constructed a mouse liver cancer model with the USP25 gene deletion to verify in vivo role of USP25. RESULTS: USP25 was highly expressed in HCC tissue and HCC cell lines. Importantly, high expression of USP25 in tissues was closely related to a poor prognosis. USP25 knockdown markedly reduced the proliferation, migration, and invasion of HepG2 and MHCC97H cells, whereas USP25 overexpression led to the opposite effects. In addition, we demonstrated that USP25 interacts with TRIM21 to regulate the expression of proteins related to epithelial-mesenchymal transition (EMT; E-cadherin, N-cadherin, and Snail) and the Wnt/β-catenin pathway (β-catenin, Adenomatous polyposis coli, Axin2 and Glycogen synthase kinase 3 beta) and those of their downstream proteins (C-myc and Cyclin D1). Finally, we verified that knocking out USP25 inhibited tumor growth and distant metastasis in vivo . CONCLUSIONS In summary, our data showed that USP25 was overexpressed in HCC. USP25 promoted the proliferation, migration, invasion, and EMT of HCC cells by interacting with TRIM21 to activate the β-catenin signaling pathway.
Animals ; Mice ; beta Catenin/genetics* ; Carcinoma, Hepatocellular/pathology* ; Cell Line, Tumor ; Cell Movement/genetics* ; Cell Proliferation/genetics* ; Epithelial-Mesenchymal Transition/genetics* ; Gene Expression Regulation, Neoplastic ; Liver Neoplasms/pathology* ; Ubiquitin Thiolesterase/metabolism* ; Wnt Signaling Pathway/genetics*

Objective:To construct a recombinant plasmid pET30a-leucine-rich repeat (LRR) containing 15 (LRRC15) of Taenia solium, prokaryotically express and purify the LRRC15 recombinant protein, and prepare a rabbit polyclonal antibody. Methods:The LRRC15 protein encoding gene of Taenia solium was obtained by whole gene synthesis; it was cloned into pET30a vector, and the recombinant plasmid pET30a-LRRC15 was constructed and identified by double-enzyme PCR; the recombinant plasmid was transformed into competent cells of Escherichia coli BL21 (DE3), and the recombinant protein LRRC15 was induced to express by isopropyl-beta-D-thiogalactopyranoside (IPTG), the expression product was analyzed and identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE); the LRRC15 recombinant protein was purified by Ni-IDA affinity columns, the purified recombinant protein was analyzed and identified by SDS-PAGE, and the specificity of the purified recombinant protein was identified by Western blot (WB); the New Zealand rabbits were immunized with purified LRRC15 recombinant protein to prepare polyclonal antibodies against LRRC15, and the potency of the purified polyclonal antibody was determined by indirect enzyme-linked immunosorbent assay (ELISA). Results:After PCR identification, a band with a length of 1 506 bp was amplified, which was consistent with the LRRC15 gene; after SDS-PAGE and WB identification, the LRRC15 target protein with a relative molecular mass ( Mr) of about 55.36 × 10 3 was obtained; after immunizing New Zealand rabbits with purified LRRC15 recombinant protein, a polyclonal antibody against LRRC15 was obtained, and its potency was 1∶1 587 200. Conclusion:The recombinant plasmid pET30a-LRRC15 is successfully constructed, the LRRC15 recombinant protein of Taenia solium is prepared, and a high purity and high potency rabbit anti polyclonal antibody against LRRC15 recombinant protein is obtained.

Centenarians are known as the "model population" in the study of healthy aging. Understanding their characteristics of health and longevity and its related influencing factors would facilitate the exploration of the possible path to achieve health and longevity under the background of population aging in China. In recent years, the population based observational studies, including China Hainan Centenarian Cohort Study (CHCCS), have found the main health characteristics of centenarians and their relationship with quality of life. This paper summarizes the relevant research results from CHCCS in recent years, and compare them with the results of similar domestic studies. The main health indicators include blood pressure, blood lipid, blood glucose, hemoglobin, body measurement and renal function. The health outcomes including diet, nutrition, health-related quality of life, activities of daily living, geriatric syndrome. The results are expected to provide epidemiological evidence for the development of healthy aging policies.

Objective:To explore the association between parathyroid hormone (PTH) and all-cause death in the centenarians in Hainan Province.Methods:The study was based on China Hainan Centenarian Cohort Study, a total of 1 002 centenarians were included in the baseline survey, the follow up was conducted to collect the information about their death outcome. The association between baseline PTH level and all-cause death was analyzed by Cox proportional risk regression model using continuous variables, dichotomous variables, and quad variables as independent variables of the centenarians.Results:As of May 31 st, 2021, a total of 522 centenarians had died in the follow up period, the median of the follow up time was 4.16 (1.31,5.04) years, and the mortality rate was 52.10%. After fully adjusting the covariates, the multiple Cox proportional hazards regression analyses showed that the risk of all-cause death increased by 3.7% for every 10 pg/ml increase in PTH value, the HR was 1.037 (95% CI: 1.007-1.067) in all the centenarians. The risk of death in the centenarians in the highest PTH group was 1.458 (95% CI: 1.131-1.878) times higher than that in the lowest PTH group. Conclusions:Higher parathyroid hormone level was associated with the elevated risk for all-cause death in the centenarians in Hainan. PTH can be used as a reference index for the prediction of the death risk of long-lived elderly in community.

Objective:To examine the association between depression and all-cause death in centenarians in Hainan Province.Methods:Based on China Hainan Centenarian Cohort Study, 1 002 centenarians with integral data were included in the baseline survey, follow up was conducted to collect the information about their survival status and death outcome. Depression was evaluated by the 15-item Geriatric Depression Scale (GDS-15) and based on the evaluation scores, the mental status of the centenarians was classified as: severe depressive disorder (MDD; score ≥10), minor depressive disorder (MnDD; score 6-9), and normal status (score ≤5). Multivariable-adjusted Cox proportional hazards regression models was used to estimate the hazard ratios ( HRs) and 95% CIs. Results:As of May 31 st, 2021, a total of 522 deaths had been reported, with a mortality density of 152.39/1 000 person-years and a median follow up time M( Q1, Q3) of 4.16 (1.31, 5.04) years, in the centenarians. After adjusting the factors of socio-demographic characteristics and lifestyle, multivariate Cox regression analysis showed that the risk of all-cause mortality increased by 3.1% for every 1-point increase in GDS-15 score ( HR=1.031, 95% CI: 1.002-1.061). Compared with centenarians with normal status, the risk of mortality in the centenarians with MDD increased by 35.9% ( HR=1.359, 95% CI: 1.023-1.806), the centenarians with MDD had an average 1.94 years of life lost. Among the female centenarians, the risk of mortality in MnDD group and MDD group increased by 29.3% ( HR=1.293, 95% CI: 1.053-1.590) and 42.3% ( HR=1.423, 95% CI: 1.047-1.935) compared with normal group, those with MDD had an average 1.71 years of life lost. Conclusions:Depression was related to the increased risk of all-cause death and life lost in the centenarians in Hainan.

Objective:To describe and analyze the relationship between traditional and new obesity related anthropometric indicators and all-cause death in centenarians in Hainan, the traditional and new obesity related anthropometric indicators included BMI, waist circumference (WC), waist hip ratio (WHR), waist height ratio, calf circumference (CC), waist-calf ratio (WCR), lipid accumulation product (LAP), visceral adiposity index (VAI), Chinese visceral adiposity index (CVAI), a body shape index (ABSI), a body shape index of Chinese (CABSI) and body roundness index.Methods:A total of 1 002 centenarians in Hainan were selected by cluster sampling. The M( Q1,Q3) follow-up time was 4.16 (1.31, 5.04) years and the outcome was all-cause death. Multiple Cox regression model was used to analyze the relationship between the obesity related anthropometric indicators and all-cause death, and the area under receiver operating characteristic curve was used for comparison. Results:In all the centenarians, CC had the strongest power to predict death, with area under curve (AUC) of 0.61 (95% CI:0.57-0.64), the gender specific results were consistent with that in overall population ( P<0.05), followed by WCR with AUC of 0.58, and then BMI, LAP and WC with AUC of 0.55, 0.55 and 0.54 respectively, while CABSI, WHR and VAI had the weaker power to predict death with AUC of 0.51, 0.50 and 0.50 respectively. Conclusions:This was the first study to prospectively compare and analyze the association between 12 obesity related anthropometric indicators and all-cause death in a large sample cohort of centenarians in China. It was found that CC had the best prediction power for death, and the risk for death decreased with the increase of CC value in a dose-response manner. It is suggested that CC can be used as a reference index for death risk monitoring in the elderly.

Objective:To understand the associations of various levels of systolic and diastolic blood pressure levels and related factors with all-cause death in the centenarians in Hainan Province.Methods:The data were obtained from China Hainan Centenarian Cohort Study, a prospective cohort study which included 1 002 centenarians. Cox proportional risk regression models were used to analyze the associations of blood pressure and related factors with all-cause death in the centenarians.Results:As of May 31, 2021, with the median of the follow-up time of 4.16 years, and the mortality density had reach 15.24 per 100 person-years. After grouping by SBP and DBP quartiles, the Q1 of SBP and Q2 of DBP caused the lowest mortality density (12.73 per 100 person-years and 14.10 per 100 person-years relatively). In all the centenarians, compared with the Q1 of SBP (≤137 mmHg, 1 mmHg=0.133 kPa), Q2, Q3 and Q4 were all positively associated with all-cause death. In the female population, the HRs of SBP's Q2, Q3 and Q4 were 1.343, 1.355 and 1.420 respectively ( Q1 as reference), and the HRs of DBP's Q1, Q3 and Q4 were 1.468, 1.800 and 1.504 respectively ( Q2 as reference), showing a roughly U-shaped association. Interactive analysis showed multiplicative interaction between SBP and BMI. Conclusions:In the centenarians in Hainan, different levels of systolic and diastolic blood pressure were associated with all-cause death. In the centenarians, DBP might have a suitable level or range in which the risk of death is relatively low. Different BMI status might affect the associations of blood pressure levels with death outcomes.