ObjectiveTo investigate the mechanism of salt processing of Psoraleae Fructus （PF） through modern analytical techniques and biotechnology， focusing on its effects related to hepatotoxicity and anti-osteoporosis activity. MethodsThe zebrafish model was utilized to evaluate the impact of PF and salt-processed Psoraleae Fructus （SPF） on the hepatotoxicity （using 134.17 ， 178.89， 268.34 mg·L^-1 as low， medium， and high dose groups of PF， 135.04， 180.06， 270.08 mg·L^-1 as low， medium， and high dose groups of SPF， respectively） and anti-osteoporotic activity （using 33.54 ， 67.08 and 134.17 mg·L^-1 as low， medium， and high dose groups of PF， 33.76， 67.52， 135.04 mg·L^-1 as low， medium， and high dose groups of SPF， respectively）， which was using alizarin red skull staining of zebrafish as an indicator of different batches of PF. The specific dosage of a batch of PF was taken as an example. Then ultra-performance liquid chromatography-quadrupole-time of flight-mass spectrometry（UPLC-Q-TOF-MS） analysis was employed to identify the chemical composition of PF before and after salt processing， and PCA， OPLS-DA， and independent sample t-test were used to elucidating the compositional changes associated with the effects of salt processing on hepatotoxicity and anti-osteoporosis activity. ResultsUnder specific conditions， PF induced notable hepatotoxicity in zebrafish while simultaneously demonstrating protective effect against prednisolone-induced osteoporosis. In comparison to PF， SPF showed alleviated hepatotoxicity while retaining significant anti-osteoporosis activity. UPLC-Q-TOF-MS analysis revealed that after salt processing， the overall chemical composition of PF showed a downward trend， with 69 components showing a decrease in content， represented by psoralen， and 13 components showing an increase， represented by 4′-O-methyl psoralen B. Further multivariate statistical analysis revealed 11 key differential components before and after salt processing of PF， including psoralen and bakuchiol. ConclusionSalt processing effectively diminishes hepatotoxicity without impairing therapeutic efficacy against osteoporosis of PF， which may be related to the compositional changes before and after salt processing of PF and provides key evidence to reveal the scientific significance of salt processing of PF.

Objective:To study the implementation progress of the prevention and control measures for drinking water-borne endemic fluorosis and the changing trend of fluorosis conditions in Inner Mongolia Autonomous Region, comprehensively evaluate the effectiveness of prevention and control measures, and provide a basis for the next step of prevention and control of the disease.Methods:In 2015 and 2023, a cross-sectional survey method was used to investigate the water improvement situation, the operation of water improvement projects, the detection of fluoride level in water, and the prevalence of dental fluorosis in children aged 8 to 12 in all affected villages of 11 leagues (cities) in the entire autonomous region. The prevention and control effect of drinking water-borne endemic fluorosis in the entire autonomous region was evaluated.Results:The water improvement rates of all affected villages in the entire autonomous region in 2015 and 2023 were 84.12% (8 218/9 769) and 95.38% (8 944/9 377), respectively. The qualified rates of water fluoride in the villages with water improvement in the entire autonomous region were 66.21% (5 441/8 218) and 91.00% (8 139/8 944), respectively. The water improvement rate and water fluoride qualification rate of water improvement villages in 2023 were both higher than those in 2015, and the differences were statistically significant (χ 2 = 652.96, 1 593.81, P < 0.001). The detection rates of dental fluorosis in children aged 8 to 12 years in the entire autonomous region in 2015 and 2023 were 9.26% (7 548/81 484) and 4.46% (3 441/77 155), respectively. The detection rate of dental fluorosis in children in 2023 was lower than that in 2015, and the difference was statistically significant (χ 2 = 1 418.20, P < 0.001). In 2015 and 2023, the total compliance rate of all affected villages in the entire autonomous region reaching the control standards were 57.94% (5 660/ 9 769) and 92.37% (8 662/9 377), respectively. The total compliance rate of all affected villages in 2023 was higher than that in 2015, and the difference was statistically significant (χ 2 = 3 010.38, P < 0.001). Conclusions:Compared with 2015, the prevention and control measures of drinking water-borne endemic fluorosis in Inner Mongolia Autonomous Region have been effectively implemented in 2023, with a significant decrease in the detection rate of dental fluorosis in children. However, there are still a few affected villages that have not achieved the control standards, and prevention and control work still need to be further strengthened.

Objective:To investigate the characteristics of the population of skeletal fluorosis patients in drinking-tea-borne endemic fluorosis (referred to as drinking-tea-borne fluorosis) areas in Inner Mongolia Autonomous Region (referred to as Inner Mongolia) and the influencing factors of treatment willingness, and to provide a basis for improving the prevention and control measures of drinking-tea-borne fluorosis and the treatment plan of skeletal fluorosis.Methods:From August to October 2022, a face-to-face questionnaire survey was conducted in key areas of drinking-tea-borne fluorosis in Inner Mongolia (administrative villages with an average daily intake of tea fluoride > 3.5 mg and skeletal fluorosis patients identified by the general survey of drinking-tea-borne fluorosis in Inner Mongolia in 2019), and to investigate the demographic, severity, and treatment status of patients with skeletal fluorosis, analyze the demographic characteristics of patients with skeletal fluorosis and the influencing factors of treatment willingness.Results:A total of 734 patients with skeletal fluorosis were investigated, including 543 mild cases, 125 moderate cases and 66 severe cases. The gender ratio of patients with skeletal fluorosis was 0.71 ∶ 1.00 (305/429), the age was concentrated in > 50 - 70 years old (70.57%, 518/734), the proportion of Mongolians was 94.28% (692/734), the proportion of herders was 97.68% (717/734), the educational level was mainly primary school (54.63%, 401/734), and the proportion of poor households and immigrants who had moved to their current residence was 7.08% (52/734) and 8.04% (59/734), respectively. The distribution of the severity of skeletal fluorosis in patients of different ages, genders, and educational levels was compared, and the differences were statistically significant ( P < 0.05). Fifty-three point two seven percent (391/734) of the patients had a willingness to undergo non-pharmacological treatment, of which 69.82% (273/391) had already started non-pharmacological treatment, with a treatment effectiveness rate of 73.99% (202/273). Sixty-five point two six percent (479/734) of the patients had a willingness to receive medication treatment, of which 7.31% (35/479) had already started medication treatment, with a treatment effectiveness rate of 54.29% (19/35). Zero point two seven percent (2/734) of the patients expressed a willingness to undergo surgical treatment, while no patients underwent surgical treatment. Multivariate logistic regression analysis showed that the age, ethnicity, occupation, educational level, poverty status, immigrants status, and the severity of skeletal fluorosis were all influencing factors of non-pharmacological treatment willingness ( P < 0.05). Occupation, educational level, poverty status, immigrants status, and the severity of skeletal fluorosis were all influencing factors of medication treatment willingness ( P < 0.05). Conclusions:Patients with skeletal fluorosis caused by tea drinking in Inner Mongolia are mainly from Mongolian ethnic groups, herders, middle-aged and elderly people, and those with a lower educational levels. The willingness of patients to receive treatment is influenced by various factors, and corresponding intervention measures can be formulated and taken based on these influencing factors to effectively improve the disease prevention awareness and treatment willingness of patients and the public.

Objective To explore the effect of evening primrose oil(EPO)on aortic endothelial damage in rats with polycystic ovary syndrome(PCOS),using network pharmacology and in vivo experiments.Methods The potential targets of EPO for improving aortic endothelial injury in PCOS rats were predicted by network pharmacology,and the selected core targets and renin-angiotensin signaling(RAS)pathway were verified by experiments.Fifty-eight female SD rats were divided randomly into a blank group(n=10)and a modeling group(n=48).Rats in the blank group were fed a normal diet and rats in the modeling group received a high-fat diet for 8 weeks.The PCOS model was prepared at week 6 by administration of letrozole(1 mg/(kg·d))for 21 days.Blood was taken from the tail vein after modeling and serum was collected to detect hormone levels.The model rats were then divided randomly into four groups and treated with the corresponding drugs for 6 weeks.Blood,blood vessels,and ovaries were then collected.Tissue morphology was examined by hematoxylin and eosin staining and serum levels of luteinizing hormone(LH),testosterone(T),follicle-stimulating hormone(FSH),endothelin(ET-1),and tumor necrosis factor(TNF-α)were detected by enzyme-linked immunosorbent assay(ELISA).Serum levels of nitric oxide(NO)were determined by spectrophotometry.Protein expression levels of core targets and RAS pathway-related factors were assessed by western blotting and immunohistochemistry.Results Twenty-five intersection targets of EPO and PCOS were identified by network pharmacological analysis.Kyoto encyclopedia of genes and genomes analysis showed that EPO improved vascular injury in PCOS rats via multiple pathways,including RAS.Serum levels of ET-1,FSH,LH,and T measured by ELISA were significantly decreased after EPO treatment,compared with the model group(P＜0.01).EPO significantly decreased the expression levels of Ang Ⅰ,VEGF-B,AT2R,ET-1,and TNF-α proteins in the aorta(P＜0.01)and significantly increased expression levels of Ang Ⅱ,CD31,and endothelial NO synthase proteins(P＜0.01).Conclusions EPO may ameliorate vascular endothelial injury in PCOS model rats by inhibiting the RAS signaling pathway and by overactivation of the ACE/Ang Ⅱ/AT1 axis.

ObjectiveTo investigate the characteristics of mitochondrial translational initiation factor 2 （MTIF2） gene methylation and its association with the development and progression of hepatocellular carcinoma （HCC）. MethodsMethSurv and EWAS Data Hub were used to perform the standardized analysis and the cluster analysis of MTIF2 methylation samples， including survival curve analysis， methylation signature analysis， the association of tumor signaling pathways， and a comparative analysis based on pan-cancer database. The independent-samples t test was used for comparison between two groups； a one-way analysis of variance was used for comparison between multiple groups， and the least significant difference t-test was used for further comparison between two groups. The Cox proportional hazards model was used to perform the univariate and multivariate survival analyses of methylation level at the CpG site. The Kaplan-Meier method was used to investigate the survival differences between the patients with low methylation level and those with high methylation level， and the Log-likelihood ratio method was used for survival difference analysis. ResultsGlobal clustering of MTIF2 methylation showed that there was no significant difference in MTIF2 gene methylation level between different races， ethnicities， BMI levels， and ages. The Kaplan-Meier survival curve analysis showed that the patients with N-Shore hypermethylation of the MTIF2 gene had a significantly better prognosis than those with hypomethylation （hazard ratio ［HR］=0.492， P<0.001）， while there was no significant difference in survival rate between the patients with different CpG island and S-Shore methylation levels （P>0.05）. The methylation profile of the MTIF2 gene based on different ages， sexes， BMI levels， races， ethnicities， and clinical stages showed that the N-Shore and CpG island methylation levels of the MTIF2 gene decreased with the increase in age， and the Caucasian population had significantly lower N-Shore methylation levels of the MTIF2 gene than the Asian population （P<0.05）； the patients with clinical stage Ⅳ had significantly lower N-Shore and CpG island methylation levels of the MTIF2 gene than those with stage Ⅰ/Ⅱ （P<0.05）. Clinical validation showed that the patients with stage Ⅲ/Ⅳ HCC had a significantly lower methylation level of the MTIF2 gene than those with stage Ⅰ/Ⅱ HCC and the normal population （P<0.05）. ConclusionN-Shore hypomethylation of the MTIF2 gene is a risk factor for the development and progression of HCC.

Objective To explore the correlation between gallbladder stones and small intestinal bacterial overgrowth(SIBO).Methods A retrospective analysis was conducted on the clinical data of 393 patients who attended the Department of Gastroenterology of the Sixth Medical Center of Chinese PLA General Hospital from January 2021 to September 2023.They were divided into gallbladder stones group(n=190)and control group(n=203)based on the presence of gallbladder stones.Their general clinical data,laboratory test results,and abdominal symptoms were compared.Multivariate logistic regression was used to analyze the risk factors for gallbladder stones.Additionally,the total population was divided into SIBO-positive group(n=239)and SIBO-negative group(n=154),and their clinical characteristics were analyzed by logistic regression to explore the risk factors for SIBO.Results Univariate analysis revealed that gallbladder stones group had a higher rate of age,body mass index(BMI),fasting plasma glucose(FPG),glutaminase levels,prevalence of hypertension,diabetes,coronary heart disease,non-alcoholic fatty liver disease,gallbladder polyps,and SIBO,as well as a higher prevalence of CH4-positive and H2-positive in SIBO group than control group(P＜0.05).In terms of abdominal symptoms,the incidence of bad breath(48.4%vs.35.5%),dyspepsia(38.4%vs.28.6%),abdominal pain(30.5%vs.14.8%),bloating(42.1%vs.28.6%),diarrhea(20.5%vs.7.4%),and more exhaustion(46.8%vs.34.5%)were significantly higher in gallbladder stones group than those in control group(P＜0.05).Multivariate logistic regression analysis showed that independent positive determinants for incident gallbladder stones were age,BMI,FPG,total bilirubin(TBIL),coronary heart disease,gallbladder polyps,and SIBO.Univariate analysis revealed that age,prevalence of gallbladder stones,proportion of single stones,triglycerides(TG),total cholesterol(TC),and low-density lipoprotein cholesterol(LDL-C)were significantly higher in SIBO-positive group than those in SIBO-negative group(P＜0.05).Multivariate logistic regression analysis showed that the risk factors for SIBO were age,coronary heart disease,and gallbladder stones,while the protective factor for SIBO was high-density lipoprotein cholesterol(HDL-C).Conclusion There is a significant correlation between gallbladder stones and small SIBO;interventions on related factors of gallbladder stones and small SIBO may help reduce their incidence.

Objective To explore the influencing factors of different types of small intestinal bacterial overgrowth(SIBO).Methods A total of 539 patients who were hospitalized in the Department of Gastroenterology,the Sixth Medical Center of PLA General Hospital from June 2021 to December 2021 and who underwent methane-hydrogen breath test were retrospectively selected.Based on breath test results,patients were divided into SIBO-negative group(n=300)and SIBO-positive group(n=239).The clinical data were compared between two groups.According to the specific values of breath test results,SIBO-positive patients were further divided into hydrogen-producing bacterial overgrowth(hydrogen-positive,n=103),intestinal methanogen overgrowth(methanogen-positive,n=80),and simultaneous methanogen and hydrogen-producing bacterial overgrowth(double positive,n=56)groups.Multivariate logistic regression analysis was employed to identify influencing factors of different SIBO types.Additionally,SIBO-positive patients were categorized by age into<45 years(n=23),45-60 years(n=82),60-75 years(n=124),and≥75 years(n=10)to compare SIBO positivity rates across age groups.Results The patients in SIBO-positive and double positive groups were older and had a lower body mass index(BMI)than those in SIBO-negative group,with statistically significant differences(P<0.05).Compared with the patients in SIBO-negative group,those in hydrogen-positive group showed a higher proportion of history of coronary heart disease,those in methanogen-positive group were older,and higher proportion of statin use,with statistically significant differences(P<0.05).Multivariate logistic regression analysis revealed that,among different SIBO types,a history of coronary heart disease served as an independent risk factor for hydrogen-producing bacterial overgrowth(OR=2.728,95%CI 1.271-5.857,P=0.010).For methanogen overgrowth,increasing age was identified as an independent risk factor(OR=1.040,95%CI 1.009-1.063,P=0.010),while the application of statin played the role of an independent protective factor(OR=0.420,95%CI 0.236-0.754,P=0.003).As for the simultaneous overgrowth of methane-producing and hydrogen-producing bacteria,increased BMI was found to be an independent protective factor(OR=0.870,95%CI 0.786-0.964,P=0.008).In SIBO-positive group,it was found that for patients aged<45 years,both the methane-positive rate and the double-positive rate were significantly lower than the hydrogen positivity rate(P<0.05).Moreover,among patients aged 45-60 years,the double-positive rate was significantly lower than the hydrogen positivity rate(P<0.01).When it comes to the hydrogen-positive rate,it was significantly lower for patients aged 45-60 and 60-75 years compared with that of patients aged<45 years(P<0.05).In contrast,the methane-positive rate and the double-positive rate were significantly higher for patients aged 45-60 and 60-75 years than those of patients aged<45 years(P<0.01).Conclusion A history of coronary heart disease and increasing age are independent risk factors for intestinal hydrogen-producing bacterial overgrowth and methanogen overgrowth,respectively.The application of statins and increased BMI are independent protective factors for intestinal methanogen simultaneous overgrowth of methanogen and hydrogen-producing bacteria,respectively.

Objective To investigate the mechanism of hippocampal neuronal plasticity of newborn neurons in the hippocampus by which exercise improves the fear and anxiety symptoms of post-traumatic stress disorder(PTSD).Methods Totally 40 C57BL/6J male mice were randomly divided into by control group(Ctrl)and PTSD group,the PTSD group was divided into a no-exercise group(PTSD),a low-intensity exercise group(L)and a high-intensity exercise group(H).The PTSD model mice were constructed by combining conditioned plantar-foot shock(CF)and single-session sustained stress(SPS).After the exercise intervention,the fear and anxiety levels of the mice were assessed using the conditioned fear test and the elevated cross maze test;Subsequently,the densities of the newborn mature neurons in dentate gyrus(DG)of hippocampus were detected by immunofluorescent double-labelling staining,and the newborn neuron morphology was marked by injecting retrovirus pRetro-U6-EF1-EGFP-3xFLAG-WPRE in DG of hippocampus to observe its morphology.The morphology of the newborn neurons was labelled to observe their dendritic length and the number of branch points;Meanwhile,the concentration level of adiponctin(APN)in the hippocampal area was determined by ELISA.Results The result showed that both high and low-intensity exercise interventions significantly reduced the freezing time of PTSD mice in the conditioned fear test,and in the elevated cross maze experiment,the residence time and the number of entries in the open arm of the mice in the H group increased significantly compared with those in the PTSD group,while the residence time and the number of entries in the closed arm were significantly reduced.In addition,both high and low-intensity exercise interventions significantly increased the surface density and dendritic length of newborn mature neurons in the hippocampal DG region of PTSD mice,and high-intensity exercise significantly increased the number of dendritic branching points,and the density of newborn mature neurons in the H group was more significantly increased compared with that in the L group.At the same time,the hippocampal APN concentration increased significantly in both L and H groups compared with the PTSD group,and it was more significant in the H group.Conclusion Exercises have an ameliorative effect on anxiety and fear symptoms in PTSD mice,and at the same time,it can increase hippocampal neuroplasticity and adiponctin secretion in PTSD mice,suggesting that the improvement of fear and anxiety symptoms in PTSD by exercise may be related to the increase of hippocampal neuroplasticity and APN secretion,and the improvement effect is better with high-intensity exercise.

Objective To explore the effect of evening primrose oil(EPO)on aortic endothelial damage in rats with polycystic ovary syndrome(PCOS),using network pharmacology and in vivo experiments.Methods The potential targets of EPO for improving aortic endothelial injury in PCOS rats were predicted by network pharmacology,and the selected core targets and renin-angiotensin signaling(RAS)pathway were verified by experiments.Fifty-eight female SD rats were divided randomly into a blank group(n=10)and a modeling group(n=48).Rats in the blank group were fed a normal diet and rats in the modeling group received a high-fat diet for 8 weeks.The PCOS model was prepared at week 6 by administration of letrozole(1 mg/(kg·d))for 21 days.Blood was taken from the tail vein after modeling and serum was collected to detect hormone levels.The model rats were then divided randomly into four groups and treated with the corresponding drugs for 6 weeks.Blood,blood vessels,and ovaries were then collected.Tissue morphology was examined by hematoxylin and eosin staining and serum levels of luteinizing hormone(LH),testosterone(T),follicle-stimulating hormone(FSH),endothelin(ET-1),and tumor necrosis factor(TNF-α)were detected by enzyme-linked immunosorbent assay(ELISA).Serum levels of nitric oxide(NO)were determined by spectrophotometry.Protein expression levels of core targets and RAS pathway-related factors were assessed by western blotting and immunohistochemistry.Results Twenty-five intersection targets of EPO and PCOS were identified by network pharmacological analysis.Kyoto encyclopedia of genes and genomes analysis showed that EPO improved vascular injury in PCOS rats via multiple pathways,including RAS.Serum levels of ET-1,FSH,LH,and T measured by ELISA were significantly decreased after EPO treatment,compared with the model group(P＜0.01).EPO significantly decreased the expression levels of Ang Ⅰ,VEGF-B,AT2R,ET-1,and TNF-α proteins in the aorta(P＜0.01)and significantly increased expression levels of Ang Ⅱ,CD31,and endothelial NO synthase proteins(P＜0.01).Conclusions EPO may ameliorate vascular endothelial injury in PCOS model rats by inhibiting the RAS signaling pathway and by overactivation of the ACE/Ang Ⅱ/AT1 axis.

Objective:To study the implementation progress of the prevention and control measures for drinking water-borne endemic fluorosis and the changing trend of fluorosis conditions in Inner Mongolia Autonomous Region, comprehensively evaluate the effectiveness of prevention and control measures, and provide a basis for the next step of prevention and control of the disease.Methods:In 2015 and 2023, a cross-sectional survey method was used to investigate the water improvement situation, the operation of water improvement projects, the detection of fluoride level in water, and the prevalence of dental fluorosis in children aged 8 to 12 in all affected villages of 11 leagues (cities) in the entire autonomous region. The prevention and control effect of drinking water-borne endemic fluorosis in the entire autonomous region was evaluated.Results:The water improvement rates of all affected villages in the entire autonomous region in 2015 and 2023 were 84.12% (8 218/9 769) and 95.38% (8 944/9 377), respectively. The qualified rates of water fluoride in the villages with water improvement in the entire autonomous region were 66.21% (5 441/8 218) and 91.00% (8 139/8 944), respectively. The water improvement rate and water fluoride qualification rate of water improvement villages in 2023 were both higher than those in 2015, and the differences were statistically significant (χ 2 = 652.96, 1 593.81, P < 0.001). The detection rates of dental fluorosis in children aged 8 to 12 years in the entire autonomous region in 2015 and 2023 were 9.26% (7 548/81 484) and 4.46% (3 441/77 155), respectively. The detection rate of dental fluorosis in children in 2023 was lower than that in 2015, and the difference was statistically significant (χ 2 = 1 418.20, P < 0.001). In 2015 and 2023, the total compliance rate of all affected villages in the entire autonomous region reaching the control standards were 57.94% (5 660/ 9 769) and 92.37% (8 662/9 377), respectively. The total compliance rate of all affected villages in 2023 was higher than that in 2015, and the difference was statistically significant (χ 2 = 3 010.38, P < 0.001). Conclusions:Compared with 2015, the prevention and control measures of drinking water-borne endemic fluorosis in Inner Mongolia Autonomous Region have been effectively implemented in 2023, with a significant decrease in the detection rate of dental fluorosis in children. However, there are still a few affected villages that have not achieved the control standards, and prevention and control work still need to be further strengthened.