Objective:To discuss the protective effect of velvet antler peptide(VAP)in the osteoporosis(OP)model rats,and to clarify the possible mechanism.Methods:Sixty 12-week-old SD rats were randomly divided into control group,model group,positive drug group(treated with 1 mg·kg-1·d-1 of alendronate sodium by gavage),low dose of VAP group(treated with 100 mg·kg-1·d-1 VAP),medium dose of VAP group(treated with 200 mg·kg-1·d-1 VAP),and high dose of VAP group(treated with 300 mg·kg-1·d-1 VAP),and there were ten rats in each group.Except for control group,the rats in the other groups were injected with dexamethasone(2 mg·kg-1)to replicate the OP rat model,while the rats in control group were injected with the equivalent volume of saline twice a week for 11 consecutive weeks.Dual-energy X-ray absorptiometry was used to detect the bone mineral density(BMD)of femur tissue of the rats in various groups;enzyme-linked immunosorbent assay(ELISA)method was used to detect the levels of serum calcium(Ca2+),phosphate(P),osteoprotegerin(OPG),alkaline phosphatase(ALP),and osteocalcin(OCN)in serum of the rats in various groups;biochemical method was used to detect the malondialdehyde(MDA)level and superoxide dismutase(SOD)activity in serum of the rats in various groups;HE staining was used to observe the pathomorphology of bone tissue of the rats in various groups;Western blotting method was used to detect the expression levels of silent information regulator 1(SIRT1),catalase(CAT),Runt-related transcription factor 2(RUNX2),and forkhead box protein O1(FOXO1)proteins in bone tissue of the rats in various groups.Results:Compared with control group,the BMD of femoral tissue of the rats in model group was decreased(P<0.05);compared with model group,the BMD of femur tissue of the rats in positive drug group,medium dose of VAP group,and high dose of VAP group were increased(P<0.05 or P<0.01).Compared with control group,the levels of Ca2+,P,OPG,and SOD activities in serum of the rats in model group were decreased(P<0.05),and the levels of ALP,OCN,and MDA were increased(P<0.05);compared with model group,the level of OPG in serum of the rats in low dose of VAP group was significantly increased(P<0.05),the levels of Ca2+,P,OPG,and activities of SOD in serum of the rats in positive drug group,medium dose of VAP group,and high dose of VAP group were significantly increased(P<0.05 or P<0.01),and the levels of ALP,OCN,and MDA in serum of the rats in positive drug group and different doses of VAP groups were decreased(P<0.05 or P<0.01).The HE staining results showed that compared with control group,the rats in model group had fewer bone cells and disordered arrangements in the bone tissue,thinner bone trabeculae with large fractures,and an expanded marrow cavity;compared with model group,the rats in positive drug group,medium dose of VAP group,and high dose of VAP group had thicker bone trabeculae arranged more tightly.The Western blotting results showed that compared with control group,the expression levels of SIRT1,CAT,RUNX2,and FOXO1 proteins in bone tissue of the rats in model group were decreased(P<0.05);compared with model group,the expression levels of SIRT1,CAT,RUNX2,and FOXO1 proteins in bone tissue of the rats in positive drug group,medium dose of VAP group,and high dose of VAP group were significantly increased(P<0.05 or P<0.01).Conclusion:VAP has the protective effect against OP in the rats,and its mechanism may be related to mediating the antioxidant stress action through the SIRT1/FOXO1 signaling pathway.

Objective     To evaluate the risk factors for postoperative in-hospital mortality in elderly patients receiving cardiac valvular surgery, and develop a new prediction models using the least absolute shrinkage and selection operator (LASSO)-logistic regression. Methods     The patients≥65 years who underwent cardiac valvular surgery from 2016 to 2018 were collected from the Chinese Cardiac Surgery Registry (CCSR). The patients who received the surgery from January 2016 to June 2018 were allocated to a training set, and the patients who received the surgery from July to December 2018 were allocated to a testing set. The risk factors for postoperative mortality were analyzed and a LASSO-logistic regression prediction model was developed and compared with the EuroSCOREⅡ. Results     A total of 7 163 patients were collected in this study, including 3 939 males and 3 224 females, with a mean age of 69.8±4.5 years. There were 5 774 patients in the training set and 1 389 patients in the testing set. Overall, the in-hospital mortality was 4.0% (290/7 163). The final LASSO-logistic regression model included 7 risk factors: age, preoperative left ventricular ejection fraction, combined coronary artery bypass grafting, creatinine clearance rate, cardiopulmonary bypass time, New York Heart Association cardiac classification. LASSO-logistic regression had a satisfying discrimination and calibration in both training [area under the curve (AUC)=0.785, 0.627] and testing cohorts (AUC=0.739, 0.642), which was superior to EuroSCOREⅡ. Conclusion     The mortality rate for elderly patients undergoing cardiac valvular surgery is relatively high. LASSO-logistic regression model can predict the risk of in-hospital mortality in elderly patients receiving cardiac valvular surgery.

Objective:To explore the clinical characteristics and risk factors of abnormal urinary albumin/creatinine ratio(UACR) in obese population.Methods:Baseline data from 2011 to 2012 in Henan Sub-center of"Risk Evaluation of cAncers in Chinese diabeTic Individuals: A lONgitudinal(REACTION) study"were utilized and those of body mass index≥28 kg/m 2 were screened. The patients were divided into UACR normal group and UACR abnormal group(101 pairs) upon being matched on a 1∶1 basis by age and gender. Multivariate logistic regression analysis, receiver operating characteristic(ROC) curve, and restricted cubic spline(RCS)analysis were performed to explore the risk factors for abnormal UACR. Results:Compared with the normal UACR group, the UACR abnormal group had a higher number of alcohol consumers, a higher prevalence of hypertension, elevated systolic blood pressure, and triglyceride(all P<0.05). Multivariate logistic regression analysis showed that alcohol consumption( P=0.008), systolic blood pressure( P<0.001), triglyceride( P=0.049), and homeostasis model assessment for insulin resistance(HOMA-IR, P=0.033) were independent risk factors for abnormal UACR in obese people. The ROC curve analysis indicated that systolic blood pressure had the strongest diagnostic performance as a single factor(ROC curve area=0.801), and there was no significant difference in diagnostic performance compared to multiple factors combination. RCS analysis results showed that the probability of abnormal UACR increased monotonically with the increase of systolic blood pressure when the systolic blood pressure was between 130 and 158 mmHg(1 mmHg=0.133 kPa). When systolic blood pressure was not in the interval, the probability of abnormal UACR did not change significantly. The results of regression analysis of triglyceride subgroup showed that when triglyceride level was greater than or equal to 5.6 mmol/L, the risk of abnormal UACR level was significantly increased( P=0.029). Conclusion:Systolic blood pressure, triglyceride, HOMA-IR, and alcohol drinking history are independent risk factors for abnormal UACR in obese people. When systolic blood pressure is≥130 mmHg or triglyceride is≥5.6 mmol/L, the risk of abnormal UACR is significantly increased.

Objectives:To evaluate and predict the risk for in-hospital mortality in elderly patients receiving cardiac valvular surgery preferably,we developed a new prediction model using machine learning. Methods:Clinical data including baseline characteristics,peri-operative data and primary endpoint of 7 163 elderly patients aged 65 years or older undergoing cardiac valvular surgery from January 2016 to December 2018 from 87 hospitals were collected from the Chinese Cardiac Surgery Registry(CCSR).Patients from January 2016 to June 2018 were assigened to the training cohort(n=5 774)and patients from July to December 2018 were assigened to the validation cohort(n=1 389).The primary endpoint was in-hospital mortality.Machine learning algorithms were used to analyze risk factors and develop prediction model. Results:Overall in-hospital mortality was 4.1%.Linear discriminant analysis(LDA),support vector classification(SVC)and logistic regression(LR)models in the training cohort all have high AUCs and low Brier scores,with good discrimination and calibration.In validation cohort,the AUC of LDA,SVC and LR were 0.744,0.744 and 0.746 respectively,which were significantly better than that of 0.642 using the European System for Cardiac Operative Risk Evaluation II(EuroSCORE II)model(P<0.05). Conclusions:The mortality rate for elderly patients undergoing cardiac valvular surgery is relatively high.LDA,SVC and LR can predict the risk for in-hospital mortality in elderly patients receiving cardiac valvular surgery with high accuracy.

Objective: To investigate the incidence trend of diabetes among children and adolescents in Ningbo City, Zhejiang Province from 2011 to 2021, so as to provide the basis for the prevention and control of diabetes among children and adolescents. Methods: Data on diabetes incidence among children and adolescents aged 0 to 18 years in Ningbo City were collected through the Diabetes Monitoring Platform of the Ningbo Chronic Disease Collaborative Management System from 2011 to 2021. Crude incidence rates were calculated and standardized using data from the Sixth National Population Census in 2010. The trend of incidence rates were analyzed by average annual percent change (AAPC). Results: A total of 701 cases of diabetes among children and adolescents were reported in Ningbo City from 2011 to 2021. The crude and standardized incidence rates were 6.86/105 and 7.27/105, respectively, showing upward trends (AAPC=5.886%, 7.147%, both P<0.05). The crude and standardized incidence rates of type 1 diabetes mellitus were 3.36/105 and 3.35/105, respectively, with no significant trend observed (AAPC=1.229%, 1.449%, both P>0.05). The crude incidence rate was higher in children and adolescents aged 10 to <15 years (4.56/105) than in other age groups (all P<0.05). The standardized incidence rate was higher in females than in males (3.49/105 vs. 3.04/105, P<0.05). The standardized incidence rate was higher in urban areas than in rural areas (3.60/105 vs. 3.15/105, P<0.05). The crude and standardized incidence rates of type 2 diabetes mellitus were 3.43/105 and 3.87/105, respectively, showing upward trends (AAPC=4.904%, 7.579%, both P<0.05). The crude incidence rate was higher in children and adolescents aged 15 to 18 years (10.53/105) than in other groups (all P<0.05). The crude incidence rates in children and adolescents who aged 10 to <15 years and 15 to 18 years showed upward trends (AAPC=15.030%, 6.637%, both P<0.05). The standardized incidence rate was higher in males than in females (4.01/105 vs. 3.57/105, P<0.05). The standardized incidence rate was higher in urban areas than in rural areas (4.57/105 vs. 3.34/105, P<0.05). Conclusions From 2011 to 2021, the incidence of type 2 diabetes mellitus showed an upward trend, with cases mainly concentrated in children and adolescents aged 15 to 18 years, males, and those living in urban areas. The incidence of type 1 diabetes mellitus remained stable, with cases mainly concentrated in children and adolescents aged 10 to <15 years, females, and those living in urban areas.

Objective: To investigate the factors affecting social alienation among elderly stroke patients, so as to provide the reference for the prevention of social alienation among elderly patients with chronic disease. Methods: The stroke patients aged ≥60 years who visited Hangzhou Hospital of Traditional Chinese Medicine from January to June 2024 were selected as subjects. Demographic information and disease status were collected through questionnaire surveys. Social alienation, stigma and social support were assessed using the General Alienation Scale, Stroke-Specific Stigma Scale and Perceived Social Support Scale, respectively. Factors affecting social alienation among elderly stroke patients were analyzed by using a multiple linear regression model. Results: Totally 283 elderly stroke patients were surveyed, including 165 males (58.30%) and 118 females (41.70%). The mean age was (69.55±8.72) years. The mean scores for social alienation, stigma and social support was (41.72±6.10), (43.70±8.96) and (59.86±10.22) points, respectively. Multiple linear regression analysis identified educational level (high school/technical secondary school, β' =-0.157; college degree or above, β' =-0.322), household monthly income per capita (≥3 000 yuan, β' =-0.260), number of stroke occurrences (recurrent stroke, β' =0.181), number of comorbid chronic diseases (≥2, β' =0.165), self-care ability (partially self-care, β' =0.142; unable to self-care, β' =0.308), stigma (β' =0.277) and social support (β' =-0.253) as factors affecting social alienation among elderly stroke patients. Conclusion Social alienation among elderly stroke patients is associated with educational level, household monthly income per capita, number of stroke occurrences, number of comorbid chronic diseases, self-care ability, stigma and social support.

Objective To investigate the pathological damage to and inflammation of different intestinal segments in a rat model of severe hemorrhage,and to explore the effect of polarization of intestinal macrophage on the pathophysiology of intestinal inflammation.Methods Male Wistar rats were randomly divided into two groups:the sham operation group and hemorrhage group.In the hemorrhage group,40％of the total blood volume was lost in 25-30 minutes,while in the sham operation group,only the femoral artery and vein were intubated without bleeding.The rats were killed at 0,3,6,12 and 24 hours.The entire intestine was isolated quickly,and sections of the intestine were cut at the duodenum,jejunum,ileocecal junction,colon and rectum for histopathological evaluation.ELISA was adopted to determine related inflammation factors while multi-color immunohistochemistry was used to calculate macrophage surface markers.The data was statistically analyzed.Results(1)Compared with the sham group,there was no significant difference in colon histology at 3 h and 6 h,but significant difference was detected in rectum scores only at 24 h.The scores of other intestinal segments were significantly different at each time point.The severity of ileocecal and colonic lesions after bleeding increased with time.The duodenum,jejunum and ileocecum were more critically injured at 3 h than the rectum at 6 h.The injury to the duodenum,jejunum,ileum and colon was much more pronounced than to the rectum at 12 h.(2)The expressions of TNF-α and IL-1β in the rectum were increased significantly at 12 h post operation.The expressions of IL-1β,TNF-α in the jejunum increased obviously at 3 h and 6 h,respectively.(3)Three hours after severe bleeding,the level of macrophages in the jejunum and ileocececal area increased significantly,and the percentage of M1 macrophages was higher.After 6 hours,the proportion of M2 macrophages in the jejunum and M1 macrophages decreased significantly.After 3 hours,the percentage of M1 macrophages in the colon decreased,but that of M2 macrophages increased.The proportion of M2 polarized macrophages in the duodenum and rectum increased at 3 h after severe bleeding but decreased at 6 h.Conclusion Pathological damage to intestinal sections after bleeding varies depending on the time,and is correlated with the inflammatory level of macrophages.

Objective:To investigate the role and mechanism of mitochondrial DNA N6-methyladenine (6mA) in the hippocampal neurons in vascular cognitive impairment.Methods:(1) In vivo experiments: SPF male rats were randomly divided into sham-operated group and chronic cerebral hypoperfusion (CCH) group ( n=12). CCH models in the CCH group were established by ligating bilateral carotid arteries, while rats in the sham-operated group were only bilaterally dissected without ligation. Exploratory ability was detected by open field test 50 d after modeling, cognitive function was evaluated by novel object recognition test 51-53 d after modeling, and learning and memory abilities were tested by Morris water maze 54-59 d after modeling. And then, rats were sacrificed; ATP concentration and reactive oxygen species (ROS) level in the hippocampal tissues were detected, and neuron apoptosis in the hippocampal CA1 area was detected by TUNEL. (2) In vitro experiments: HT-22 cells were divided into normal control (NC) group, oxygen-glucose deprivation (OGD) group, OGD+siControl group, and OGD+siMETTL4 group. Cells in the NC group were cultured routinely, cells in the OGD group were subjected to low sugar and low oxygen for 12 h, and cells in the OGD+siControl group and OGD+siMETTL4 group were, respectively, transfected with NC-siRNA or METTL4-siRNA after being subjected to low sugar and low oxygen for 12 h. Mitochondria morphology was observed by transmission electron microscopy, ROS was detected by flow cytometry, mitochondria membrane potential was detected by JC-1 fluorescent staining, and mitochondrial complex I and III activity was detected by kit. (3) In vivo and in vitro experiments: METTL4 and DNA 6mA expressions in neuronal mitochondria of rat hippocampal tissues and mitochondria of HT-22 cells were detected by immunofluorescent staining and Western blotting. Results:(1) CCH rats had cognitive impairment: compared with the sham-operated group, CCH group had significantly increased frequency of entering the central area and reduced time in exploring new objects in open field experiment,and significantly decreased frequency of crossing the platform and prolonged escape latency in water maze experiment ( P<0.05). Compared with rats in the sham-operated group, rats in the CCH group had significantly decreased hippocampal ATP content ([18.820±1.177] nmol/L vs. [10.190±0.519] nmol/L) and increased ROS content ([4 488.00±255.70] AU vs. [11 644.00±530.20] AU, P<0.05). TUNEL results showed that the number of apoptotic neurons in the hippocampal CA1 area of CCH group was obviously increased than that in sham-operated group. Immunofluorescent staining results showed that 6mA and METTL4 mainly distributed in the mitochondria of hippocampal neurons in CCH group, and the 6mA and METTL4 expressions were obviously increased compared with those in the sham-operated group. Western blotting results showed that METTL4 expression in the hippocampal mitochondria of CCH group was significantly higher than that in the sham-operated group (1.729±0.168 vs. 1.000±0.000). (2) In vitro experiment: under transmission electron microscope, compared with the NC group, HT-22 cells in the OGD group showed obvious mitochondrial ridge disappearance, membrane rupture and vacuolation. Compared with the OGD group, the OGD+siMETTL4 group had significantly increased ATP production, decreased mtROS production, increased mitochondrial membrane potential, and increased mitochondrial complex I and III activities ( P<0.05). Immunofluorescent staining results showed that the mtDNA 6mA and METTL4 expressions in the OGD group were obviously higher than those in the NC group, and both mainly expressed in the mitochondria; mtDNA 6mA expression in the OGD+siMETTL4 group was obviously lower than that in OGD group. Western blotting results showed that METTL4 expression in the OGD+siMETTL4 group was significantly higher than that in the OGD group (1.578±0.261 vs. 2.970±0.280). Conclusion:Specific high expression of methylase METTL4 in hippocampal neurons of rats with cognitive impairment after CCH promotes the increased mtDNA 6mA expression and leads to mitochondrial energy metabolism disorders and increased ROS, which is speculated to be one of the mechanisms causing vascular cognitive impairment.

Objective:To investigate the causal relationship between intestinal flora and benign biliary diseases by genome-wide Mendelian randomization.Methods:This is a retrospective observational study. The data from the genome-wide association study of the gut microbiota from 18 340 samples from the MiBioGen consortium were selected as the exposure group,and the data from the genome-wide association study of biliary tract diseases were obtained from the FinnGen consortium R8 as the outcome group. There were 1 491 cases of primary sclerosing cholangitis,32 894 cases of cholelithiasis,3 770 cases of acalculous cholecystitis,and 34 461 cases of cholecystitis. Single nucleotide polymorphisms were screened as instrumental variables,and the Mendelian randomization method was used to infer the causal relationship between exposures and outcomes. The inverse variance weighting method (IVW) was used as the main basis, supplemented by heterogeneity,pleiotropy and sensitivity tests.Results:Coprococcus 2 was associated with a reduced risk of cholelithiasis (IVW OR=0.88,95% CI:0.80 to 0.97, P=0.012) and cholecystitis (IVW OR=0.88,95% CI:0.80 to 0.97, P=0.011). Coprococcus 3 was associated with cholelithiasis (IVW OR=1.15,95% CI:1.02 to 1.30, P=0.019) and acalculous cholecystitis(IVW OR=1.48, 95% CI: 1.08 to 2.04, P=0.016) and cholecystitis (IVW OR=1.17, 95% CI: 1.02 to 1.33, P=0.020). Peptococcus was associated with an increased risk of cholelithiasis (IVW OR=1.08, 95% CI:1.02 to 1.13, P=0.005) and cholecystitis (IVW CI=1.07, 95% CI:1.02 to 1.13, P=0.010). Clostridiumsensustricto 1 was associated with an increased risk of cholelithiasis (IVW OR=1.16,95% CI:1.02 to 1.31, P=0.020) and cholecystitis (IVW OR=1.16, 95% CI:1.03 to 1.30, P=0.015). Eubacterium hallii was associated with an increased risk of primary sclerosing cholangitis (IVW OR=1.43, 95% CI: 1.03 to 1.99, P=0.033). Eubacterium ruminantium (IVW OR=0.87, 95% CI: 0.76 to 1.00, P=0.043) and Methanobrevibacter (IVW OR=0.81, 95% CI: 0.68 to 0.98, P=0.027) were associated with a reduced risk of acalculous cholecystitis. Conclusions:Eight intestinal bacterial genera maybe play pathogenic roles in benign biliary diseases. Eubacterium hallii can increase the risk of primary sclerosing cholangitis. Peptococcus and Clostridiumsensustricto 1 can increase the risk of cholelithiasis and generalized cholecystitis. Coprococcus 3 have multiple correlations with biliary stones and inflammation.

Surgical treatment is one of the most important forms of treatment in patients with gallbladder cancer. With the development of minimally invasive technology, the feasibility, safety and efficacy of minimally invasive approaches such as laparoscopic or robotic-assisted radical cholecystectomy for gallbladder cancer have received continuous attention.For patients with an early T‐stage (Tis or T1a), laparoscopic simple cholecystectomy is safe and economical, with a good prognosis for postoperative patients, and it has been widely accepted and performed. Radical resection of advanced gallbladder cancer requires resection of the gallbladder, its liver bed, and other neighboring invaded organs, as well as clearance of regional lymph nodes, which requires experienced gallbladder cancer treatment teams to strictly grasp the indications, select appropriate patients, and formulate a good surgical strategy to ensure the therapeutic effect. Meanwhile, robot-assisted radical resection for gallbladder cancer has been performed in a few centers and shows good clinical potential, but more high-quality studies are needed to further evaluate its value in gallbladder cancer treatment.