Objective:To investigate the effect of betaine in oxygen-glucose deprivation injury of rat brain microvascular endothelial cells(BMECs),and to clarify the regulatory effect of betaine on phosphoinositide 3-kinase(PI3K)/protein kinase B(AKT)pathway.Methods:Five SD rats aged 7 d were selected and the rat BMSEs were obtained.The oxygen-glucose deprivation model of rat BMECs was prepared under hypoxic and hypoglycemic conditions;the experiment was divdided into model group,and low dose,medium dose,and high dose of betaine groups and positive control group,at the same time,blank control group(without modeling)was set up.The BMECs in blank control group and model group were treated with fresh medium,the BMECs in positive control group were given a final concentration of 10 μmnol·L-1 nimodipine,and the BMECs in low,medium and high doses of betaine groups were treated with betaine at final concentration of 0.5,1.0 and 2.0 mmol·L-1,respectively.The survival rates of BMECs in various groups were determined by CCK-8 method at 12,24 and 48 h after culture;the activities of lactate dehydrogenase(LDH)and the levels of adenine ribonucleoside triphosphate(ATP)in the rat BMECs in various groups were determined using kits,and the levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-6,IL-1β,and IL-18 in supernatants of the BMECs in various groups were determined by enzyme-linked immunosorbent assay(ELISA);the activities of superoxide dismutase(SOD)and levels of malondialdehyde(MDA)in the BMECs in various groups were determined by kits;the transendothelial resistance(TEER)values of rat BMSCs in various groups were determined by TEER analyzer,and the horseradish peroxidase(HRP)permeabilities of BMECs in various groups were determined by an insertion cell culture apparatus.TUNEL staining was used to determine the apoptotic rates of rat BMECs in vaisous groups,and Western blotting method was used to determine the ratios of phosphory lated PI3K(p-PI3K)/PI3K and phosphorylated AKT(p-AKT)/AKT in the rat BMECs in various groups.Results:Compared with blank control group,the survival rate of BMECs,activity of SOD,and level of ATP,value of TEER,and ratios of p-PI3K/PI3K and p-AKT/AKT of the rat BMECs in model group were significantly decreased(P＜0.05),while the activity of LDH,the levels of TNF-α,IL-6,IL-1β,IL-18,and MDA,the apoptotic rate of the BMECs,and HRP permeability were significantly increased(P＜0.05).Compared with model group,the survival rates of the BMECs,activities of SOD,and levels of ATP,values of TEER,and ratios of p-PI3K/PI3K and p-AKT/AKT of the BMECs in low,medium,and high doses of betaine groups and positive control group were significantly increased(P＜0.05),while the activities of LDH,the levels of TNF-α,IL-6,IL-1β,IL-18,and MDA,the apoptotic rates of BMECs and HRP permeabilities were significantly decreased(P＜0.05).Conclusion:Betaine can significantly repair the oxygen-glucose deprivation/reperfusion injury in the rat BMECs,inhibit the oxidative damage and apoptosis of BMECs,and improve the permeability of the cells;its mechanism may be related to the regulation of the PI3K/AKT pathway.

AIM:By establishing ischemic stroke(IS)rats and cell models,this study aimed to investigate the therapeutic effect of an enriched environment(EE)and to explore its impact on the neurotrophin 3(NT3)/p75 neuro-trophin receptor(p75NTR)signaling pathway.METHODS:The study consisted of in vivo and in vitro experiments.In vi-vo,Sprague-Dawley(SD)rats were randomly divided by weight into sham,IS,and IS+EE groups(n=10),with 8 addi-tional rats per group reserved for supplementary analyses.The IS model was established by the Longa suture occlusion method.Neurological and motor function deficits were assessed on days 1,3,7 and 14 post-modeling using the modified neurological severity score(mNSS).On days 3,7 and 14,4 additional rats from each group were sacrificed,and the whole-brain tissue was collected to measure infarct volume via 2,3,5-triphenyltetrazolium chloride(TTC)staining.On day 14,brain tissue was harvested for immunofluorescence staining to evaluate neuronal proliferation markers,while the ischemic penumbra was analyzed by Western blot for NT3/p75NTR pathway protein expression.In vitro,primary neural stem cells(NSCs)were isolated from fetal rats and cultured as neurospheres.These cells were divided into CON group and experimental groups treated with different concentrations of NT3 to evaluate the effects of NT3 on NSC proliferation and migration.Additionally,SH-SY5Y cell lines were used to establish an in vitro model of ischemic stroke through oxy-gen-glucose deprivation(OGD).These cells were treated with varying concentrations of NT3,along with CON and CON+NT3 groups,and a scratch assay was performed to assess the impact of NT3 on cell migration.RESULTS:EE significant-ly reduced neurological function scores in IS rats(P<0.05),prolonged latency in the rotarod test(P<0.05),and de-creased cerebral infarct area(P<0.05).EE further enhanced the protein expression of BrdU and Ki67 in the ischemic penumbra(P<0.05),as well as increased the co-expression of BrdU/DCX and BrdU/NeuN(P<0.05).Additionally,EE further upregulated the protein expression of NT3,p75NTR,PI3K,and Akt in the subventricular zone(SVZ)(P<0.05).In vitro,NT3(1 and 10 μg/L)significantly increased nestin expression(P<0.05)in the primary neural stem cell system.In the neural stem cell sphere system,compared to the CON group,1 μg/L NT3 markedly enhanced tubulin and phalloidin protein expression(P<0.05).In the scratch assay,1 ug/L NT3 significantly promoted the migration of both normal SH-SY5Y cells and OGD-induced SH-SY5Y cells compared to the CON group(P<0.05).CONCLUSION:Enriched envi-ronment activates the NT3/p75NTR signaling pathway,promoting the proliferation of NSCs in the SVZ and their migration to the ischemic penumbra,where they ultimately differentiate into neurons to replace those damaged,thereby contributing to the improvement of neurological function in rats with IS.

Objective To investigate the effect of an enriched environment on glucose metabolism in the ischemic penumbra of rats with ischemic stroke(IS).Methods SD rats were subjected to middle cerebral artery ischemia to establish an IS model.The rats were ran-domly grouped and housed under different conditions with scheduled drug administration for 21 consecutive days.Neurological function scores and TTC staining were used to assess animal IS symptoms,whereas HPLC and Western blotting were used to detect glucose me-tabolic indicators and related protein expression,respectively.Results Compared to the model group,rats in the enriched envi-ronment group showed significant improvement in IS symptoms,with increased levels of glucose metabolic indicators and related protein expression.Compared to the positive drug group,rats in the positive drug+enriched environment group showed further improvement in IS symptoms,and the levels of glucose metabolic indicators and related protein expression were further enhanced.Conclusion Enriched environment can significantly improve IS symptoms in rats,and its mechanism may be related to an increase in glucose metabolic levels.

Objective:To explore the categories and characteristics of health behaviors of community residents in Dongfang City, Hainan Province, and to analyze the influencing factors of different categories.Methods:From March to May 2023, 795 community residents of Dongfang City were selected for the study using a simple random sampling method. General Information Questionnaire, Chinese version Short of Health Promotion Lifestyle Profile-Ⅱ, Health Conception Scale, and Self-rated Abilities for Health Practice Scale were used to conduct the survey. Cluster analysis was used to explore the health behavior categories of community residents, and a decision tree model was applied to analyze the influencing factors of different categories.Results:A total of 830 questionnaires were distributed, and 795 valid questionnaires were recovered, with a valid recovery rate of 95.78%. The health behaviors of 795 community residents in Dongfang City were divided into four categories of comprehensive behavior group (26.04%, 207/795), healthy behavior group (37.23%, 296/795), risky behavior group (18.99%, 151/795), and poor health behavior group (17.74%, 141/795). The decision tree model showed that exercise efficacy, gender, ethnicity, and clinical health concepts were the factors influencing different health behavior categories of community residents in Dongfang City ( P<0.05), with exercise efficacy being the core factor. Conclusions:There is heterogeneity in the health behavior of community residents in Dongfang City. Community healthcare workers can implement targeted health promotion measures according to the behavioral characteristics of the population.

AIM:By establishing ischemic stroke(IS)rats and cell models,this study aimed to investigate the therapeutic effect of an enriched environment(EE)and to explore its impact on the neurotrophin 3(NT3)/p75 neuro-trophin receptor(p75NTR)signaling pathway.METHODS:The study consisted of in vivo and in vitro experiments.In vi-vo,Sprague-Dawley(SD)rats were randomly divided by weight into sham,IS,and IS+EE groups(n=10),with 8 addi-tional rats per group reserved for supplementary analyses.The IS model was established by the Longa suture occlusion method.Neurological and motor function deficits were assessed on days 1,3,7 and 14 post-modeling using the modified neurological severity score(mNSS).On days 3,7 and 14,4 additional rats from each group were sacrificed,and the whole-brain tissue was collected to measure infarct volume via 2,3,5-triphenyltetrazolium chloride(TTC)staining.On day 14,brain tissue was harvested for immunofluorescence staining to evaluate neuronal proliferation markers,while the ischemic penumbra was analyzed by Western blot for NT3/p75NTR pathway protein expression.In vitro,primary neural stem cells(NSCs)were isolated from fetal rats and cultured as neurospheres.These cells were divided into CON group and experimental groups treated with different concentrations of NT3 to evaluate the effects of NT3 on NSC proliferation and migration.Additionally,SH-SY5Y cell lines were used to establish an in vitro model of ischemic stroke through oxy-gen-glucose deprivation(OGD).These cells were treated with varying concentrations of NT3,along with CON and CON+NT3 groups,and a scratch assay was performed to assess the impact of NT3 on cell migration.RESULTS:EE significant-ly reduced neurological function scores in IS rats(P<0.05),prolonged latency in the rotarod test(P<0.05),and de-creased cerebral infarct area(P<0.05).EE further enhanced the protein expression of BrdU and Ki67 in the ischemic penumbra(P<0.05),as well as increased the co-expression of BrdU/DCX and BrdU/NeuN(P<0.05).Additionally,EE further upregulated the protein expression of NT3,p75NTR,PI3K,and Akt in the subventricular zone(SVZ)(P<0.05).In vitro,NT3(1 and 10 μg/L)significantly increased nestin expression(P<0.05)in the primary neural stem cell system.In the neural stem cell sphere system,compared to the CON group,1 μg/L NT3 markedly enhanced tubulin and phalloidin protein expression(P<0.05).In the scratch assay,1 ug/L NT3 significantly promoted the migration of both normal SH-SY5Y cells and OGD-induced SH-SY5Y cells compared to the CON group(P<0.05).CONCLUSION:Enriched envi-ronment activates the NT3/p75NTR signaling pathway,promoting the proliferation of NSCs in the SVZ and their migration to the ischemic penumbra,where they ultimately differentiate into neurons to replace those damaged,thereby contributing to the improvement of neurological function in rats with IS.

Objective:To explore the categories and characteristics of health behaviors of community residents in Dongfang City, Hainan Province, and to analyze the influencing factors of different categories.Methods:From March to May 2023, 795 community residents of Dongfang City were selected for the study using a simple random sampling method. General Information Questionnaire, Chinese version Short of Health Promotion Lifestyle Profile-Ⅱ, Health Conception Scale, and Self-rated Abilities for Health Practice Scale were used to conduct the survey. Cluster analysis was used to explore the health behavior categories of community residents, and a decision tree model was applied to analyze the influencing factors of different categories.Results:A total of 830 questionnaires were distributed, and 795 valid questionnaires were recovered, with a valid recovery rate of 95.78%. The health behaviors of 795 community residents in Dongfang City were divided into four categories of comprehensive behavior group (26.04%, 207/795), healthy behavior group (37.23%, 296/795), risky behavior group (18.99%, 151/795), and poor health behavior group (17.74%, 141/795). The decision tree model showed that exercise efficacy, gender, ethnicity, and clinical health concepts were the factors influencing different health behavior categories of community residents in Dongfang City ( P<0.05), with exercise efficacy being the core factor. Conclusions:There is heterogeneity in the health behavior of community residents in Dongfang City. Community healthcare workers can implement targeted health promotion measures according to the behavioral characteristics of the population.

Objective To investigate the effect of an enriched environment on glucose metabolism in the ischemic penumbra of rats with ischemic stroke(IS).Methods SD rats were subjected to middle cerebral artery ischemia to establish an IS model.The rats were ran-domly grouped and housed under different conditions with scheduled drug administration for 21 consecutive days.Neurological function scores and TTC staining were used to assess animal IS symptoms,whereas HPLC and Western blotting were used to detect glucose me-tabolic indicators and related protein expression,respectively.Results Compared to the model group,rats in the enriched envi-ronment group showed significant improvement in IS symptoms,with increased levels of glucose metabolic indicators and related protein expression.Compared to the positive drug group,rats in the positive drug+enriched environment group showed further improvement in IS symptoms,and the levels of glucose metabolic indicators and related protein expression were further enhanced.Conclusion Enriched environment can significantly improve IS symptoms in rats,and its mechanism may be related to an increase in glucose metabolic levels.

Objective : To investigate the effects and underlying mechanisms of α-mangostin in a spinal cord inj ury model of microglial cell inflammation . Methods : Mouse microglial cell line BV-2 was cultured in vitro , and an in- flammation model was established by co-treatment with lipopolysaccharide and adenosine triphosphate (LPS/ATP) . The CCK-8 assay was used to test the influence of different concentrations (0 , 10 , 20 , 40 , 80 μmol/L) of α-man- gostin on cell proliferation vitality under LPS/ATP stimulation to select an appropriate concentration range of α- mangostin; BV-2 cells were divided into Ctrl group , LPS/ATP group , 40 μmol/L α-mangostin group , and inter- vention groups with different concentrations (10 , 20 , 40 μmol/L) of α-mangostin ( designated as LPS/ATP + 10 μmol/L α-mangostin group , LPS/ATP + 20 μmol/L α-mangostin group , and LPS/ATP + 40 μmol/L α-mangostin group , respectively) . ELISA experiments were conducted to detect the levels of pro-inflammatory cytokines inter- leukin -6/1β/18 (IL-6 , IL-1β, IL-18) and tumor necrosis factor (TNF-α) in the supernatants of each group , and Western blot was used to detect the expression of NLRP3 , ASC , cleaved caspase-1 , IL-1β, and the phosphoryla- tion levels of p65 (p-p65/p65) in the NF- κB pathway , as well as the expression of p65 in the nuclei of BV-2 cells . Results : Compared with the Ctrl group, cell proliferation vitality in the LPS/ATP group was significantly reduced (P < 0. 05) , but low concentrations (10 , 20 , 40 μmol/L) of α-mangostin significantly improved the inhibi- tory effect of LPS/ATP on microglial cell proliferation vitality (P < 0. 05) , while a high concentration (80 μmol/ L) of α-mangostin exacerbated the damage to microglial cells caused by LPS/ATP (P < 0. 05) . C ompared with the Ctrl group , the levels of inflammatory factors IL-6 , IL-1β, IL-18 , TNF-α, and the expression of NLRP3 , ASC , cleaved caspase-1 , IL-1β, and the p-p65/p65 ratio in the 40 μmol/L α-mangostin group , as well as the expression of p65 protein in the nuclei , showed no significant changes ( P > 0 . 05) , whereas these significantly increased in the LPS/ATP group (P < 0. 05) . Compared with the LPS/ATP group , the levels of IL-6 , IL-1β, IL-18 , TNF-α, and the expression of NLRP3 , ASC , cleaved caspase-1 , IL-1β, and the p-p65/p65 ratio in the intervention groups , as well as the expression of p65 protein in the nuclei , decreased in a concentration-dependent manner with increasing α-mangostin concentration , with the most significant reduction ob served in the LPS/ATP + 40 μmol/L α- mangostin group (P < 0. 01) . Conclusion α-mangostin can inhibit the neuroinflammatory response mediated by NLRP3 inflammasome activation in BV-2 cells through the NF- κB pathway .

Objective To investigate the impact of anti-reflux modified duodenal exclusion surgery on glucose metabolism in rats with type 2 diabetes mellitus (T2DM), and to elucidate the role of the duodenum in maintaining glucose homeostasis. MethodsForty male Sprague-Dawley rats aged 5 weeks were fed a high-fat diet and induced with T2DM using low-dose streptozotocin. Thirty-six rats that met the T2DM model criteria were randomly divided into three groups: the simple duodenal exclusion surgery group (DE group), the anti-reflux modified duodenal exclusion group (MDE group), and the sham operation group (SO group), with 12 rats in each group. Gastroenterography was performed 4 weeks after surgery, and the body weight, fasting blood glucose levels, and serum glucagon-like peptide-1 (GLP-1) concentrations were measured before surgery and at 1, 2, 4, and 8 weeks post-surgery. Eight weeks post-surgery, the rats were euthanized, and a 1 cm segment of the biliopancreatic loop was collected from each group for pathological sectioning and HE staining to observe the intestinal mucosal villus length under an optical microscope. Results Gastroenterography showed that there was significant reflux of the contrast agent into the duodenal lumen in the DE group, while no reflux was observed in the MDE group. At one week post-surgery, the body weights of rats in all three groups significantly decreased compared to before surgery (P<0.05), and then the body weights of all groups increased over time, with no significant differences between the groups (P>0.05). Compared with the SO group, the fasting blood glucose levels in the MDE and DE groups significantly decreased at all time points post-surgery (P<0.05), while GLP-1 concentrations significantly increased (P<0.05). The fasting blood glucose levels in the MDE group were lower than those in the DE group at all time points post-surgery (P<0.05), but there were no significant differences in serum GLP-1 concentrations between the MDE and DE groups (P>0.05). Regarding intestinal mucosal morphology, the villus lengths of the biliopancreatic loops in the MDE group were significantly shorter than those in the DE and SO groups (P<0.05). Conclusion Anti-reflux modified duodenal exclusion surgery effectively improves glucose metabolism in T2DM rats by preventing the reflux of chyme into the diverted duodenum, thereby enhancing its hypoglycemic effect.

【Objective】 To investigate the frequency of platelet antibodies in voluntary blood donors and patients in Zhongshan, Guangdong Province, and to study the specificity and cross-matching of platelet antibodies. 【Methods】 Platelet antibodies of blood donors and patients were screened by solid-phase immunoadsorption (SPIA), rechecked by flow cytometry (FCM), and antibody specificity was identified by PakPlus enzyme immunoassay, and platelet cross-matching was simulated by SPIA. 【Results】 A total of 1 049 blood donor samples and 598 patient samples were tested, with 6 (0.57%) and 49 (8.19%) samples positive for SPIA,respectively(P<0.05); In SPIA positive samples, the positive concordance rate of FCM in blood donors and patients was 100% vs 95%, and that of enzyme immunoassay was 100% vs 88%. Among the initial screening positive samples of blood donors, 5 were anti-HLA Ⅰ antibodies, accounting for 83%, and 1 was anti CD36 antibody, accounting for 17%, with an incidence rate of 0.10%. Among the 14 samples of enzyme immunoassay positive patients, 2 were anti-GP Ⅱb/Ⅲa, 1 was anti-GP Ⅱa/Ⅱa, 8 were anti HLA Ⅰ, and 3 were mixed antibodies (HLA Ⅰ, GP Ⅱb/Ⅲa, GP Ⅰa/Ⅱa). According to the types of antibodies, HLA Ⅰ antibodies were the most common, accounting for 65% (11/17), followed by HPA related anti GP, accounting for 35% (6/17). The majority of patients had a platelet antibody positive typing rate below 30%, accounting for 71.4% (10/14). 【Conclusions】 The positive rate of platelet antibody of patients in Zhongshan area is significantly higher than that of voluntary blood donors, and most of them are anti-HLA Ⅰ and anti-GP, and the incidence of anti-CD36 is extremely low. Therefore, it is necessary to establish a known platelet antigen donor bank, and at the same time, carry out platelet antibody testing and matching of patients, which is helpful to solve the issue of platelet transfusion refractoriness.