Objective:To explore the factors affecting and improvement recommendations about the congealing temperature of benzyl benzoate as pharmaceutical excipient by the method in the Chinese pharmacopoeia.Methods:The influence of seven factors such as measuring device,sample size,melting temperature,condensation tempera-ture,stop stirring time,stirring frequency and reading of congealing temperature on the determination of congealing temperature of benzyl benzoate was investigated.Results:The key factor for determination of benzyl benzoate con-gealing temperature was the stop stirring time,which was more reasonable when the sample temperature drops to constant or starts to rise slightly was reasonable.Conclusion:The improved method has better accuracy and repeat-ability,and is suitable for the measuring the congealing temperature of pharmaceutical excipient benzyl benzoate.

AIM:This study examined the inhibitory effect of peiminine on the human colonic adenocarcino-ma cell line SW480 and explored the underlying mechanisms.METHODS:SW480 and human normal colonic epithelial CCD-841CoN cells were treated with different concentrations of peiminine and subjected to the CCK-8 assay to select the optimal treatment time and concentration of the compound.SW480 cell migration and invasion were evaluated by the wound-healing and Transwell assays.Cell cycle progression was analyzed by flow cytometry.The expression levels of cell cycle-related proteins were examined by Western blot.SW480 xenograft tumor model was established in nude mice to ex-amine the effect of peiminine on tumor growth and the expression of cell cycle-related proteins in vivo.RESULTS:Peimi-nine(110 mg/L)significantly inhibited the proliferation of SW480 cells compared with the control group(P<0.01),caused cell cycle arrest at G1 phase,and significantly downregulated the expression of cyclin dependent kinase 2(CDK2),CDK4,CDK6,cyclin D1,p-Rb/Rb,E2F1,E2F3,and E2F4(P<0.05).Peiminine inhibited SW480 xenograft tumor growth,prolonged the survival of model mice,and affected the expression of CDK2,CDK4,CDK6,and cyclin D1 in tu-mor tissues.CONCLUSION:Peiminine promotes G1 phase arrest by down-regulating the expression of CDK2,CDK4,CDK6,and cyclin D1,thereby inhibiting the proliferation of SW480 cells.

AIM:To explore the effect and mechanism of action of the aqueous extract of Fritillaria ussuriensis(FU-AE)against nonalcoholic fatty liver disease(NAFLD).METHODS:The association between Fritillaria ussuriensis Maxir.(FU)and NAFLD was analyzed by network pharmacology.A mouse model of NAFLD was induced in mice by high fat diet(HFD)+10%fructose drinking water,and three doses of Fritillaria ussuriensis aqueous extract were given to the mice for intervention.Colorimetric assay was used for detection of aspartate aminotransferase(AST),alanine aminotrans-ferase(ALT),triglyceride(TG),total cholesterol(TC),high-density lipoprotein cholesterol(HDL-C),and low-density lipoprotein cholesterol(LDL-C)levels in the serum of experimental mice.Hematoxylin and eosin staining was used to as-sess the pathological and histological changes in the liver of mice and to clarify the anti-NAFLD effect of aqueous extracts of Fritillaria ussuriensis.Liver tissue proteins were extracted,and expression of proteins related to the AMP-activated pro-tein kinase(AMPK)/acetyl-CoA carboxylase(ACC)pathway was detected by Western blot to clarify the mechanism of an-ti-NAFLD action of Fritillaria ussuriensis.The microbial composition of cecum contents was explored using 16S rRNA se-quencing to reveal the modulatory effect of the aqueous extract of Fritillaria ussuriensis on the structure of intestinal flora in mice with nonalcoholic fatty liver disease.RESULTS:Aqueous extract of Fritillaria ussuriensis(high dose)ameliorated exogenous adipocyte infiltration in the liver of mice with NAFLD(P<0.05).AST,ALT,TG,TC and LDL-C levels were significantly decreased(P<0.05)and HDL-C levels were significantly increased(P<0.05)in the high-dose group.Aque-ous extract of Fritillaria ussuriensis(high dose)significantly increased expression of phosphorylated AMPKα,AMPKα,and phosphorylated ACC in the livers of the model mice(P<0.05),significantly reduced expression of ACC(P<0.05),and significantly increased the relative abundance of the potentially beneficial bacteria Faecalibaculum rodentium,Lacto-bacillus johnsonii,Akkermansia muciniphila(P<0.05).CONCLUSION:Aqueous extract of Fritillaria ussuriensis may ameliorate NAFLD in mice by activating the AMPK/ACC pathway and modulating the structure of intestinal flora.

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.

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.

Objective:To explore the association between nutritional status and all-cause death in the centenarians in Hainan Province.Methods:Based on the survey data of China Hainan Centenarian Cohort Study from 2014 to 2021, a total of 1 002 Hainan centenarians with complete baseline data were included in this study, and their survival status and death outcome were surveyed. According to the Mini Nutritional Assessment Short-Form (MNA-SF), the centenarians were divided into three groups: well-nourished (12-14), at risk of malnutrition (8-11), and malnutrition (0-7). The survival status of the centenarians was evaluated by Kaplan-Meier curve. Cox proportional hazards regression model was used to assess the association between nutritional status and all-cause death.Results:After the follow-up on May 31, 2021, we had found that 522 centenarians died, with an all-cause mortality rate of 52.10% (522/1 002). Compared with the well-nourished group, the average life lost caused by malnutrition was 0.62 years. Kaplan-Meier survival analysis showed that all-cause mortality rate was higher in the malnourished centenarians than in other groups ( χ2=16.45, P<0.001). Multivariate Cox proportional hazards regression model showed that the risk of all-cause mortality rate in malnourished centenarians was higher than that in well-nourished centenarians ( HR=1.65, 95% CI: 1.18-2.31). Subgroup analysis found that the association in female centenarians was more significant. Conclusions:Malnutrition was associated with a high risk of all-cause death in Hainan centenarians. It is suggested that we should timely evaluate and pay attention to the impact of nutritional status of centenarians on their health and longevity, and death, especially in the female elderly.