BACKGROUND:Previous studies have found that neohesperidin can delay bone loss in ovariectomized mice and has the potential to treat osteoporosis,but its specific mechanism of action remains to be explored. OBJECTIVE:To explore the key targets and possible mechanisms of neohesperidin in the treatment of osteoporosis based on bioinformatics and cell experiments in vitro. METHODS:The gene expression dataset related to osteoporosis was obtained from GEO database,and the differentially expressed genes were screened and analyzed in R language.The osteoporosis-related targets were screened from GeneCards and DisGeNET databases,and the neohesperidin-related targets were screened from ChEMBL and PubChem databases,and the common targets were obtained by intersection of the three.The String database was used to construct the PPI network of intersection genes,and the key targets were screened.The DAVID database was used for GO and KEGG enrichment analysis.The AutoDock software was used to verify the molecular docking between the neohesperidin and the target protein.The effect of neohesperidin on osteogenic differentiation of C57 mouse bone marrow mesenchymal stem cells was detected.Complete medium was used as blank control group;osteogenic induction medium was used as the control group;and osteogenic induction medium containing different concentrations of neohesperidin(25,50 μmol/L)was used as experimental group.The expression of alkaline phosphatase,the degree of mineralization,the expression of osteogenic-related genes and target genes during osteogenic differentiation of cells were measured at corresponding time points. RESULTS AND CONCLUSION:(1)9 253 differentially expressed genes,2 161 osteoporosis-related targets,and 326 neohesperidin-related targets were screened.There were 53 common targets among the three.All 53 genes were up-regulated in osteoporosis samples.The PPI network screened the target gene PRKACA of research significance.GO function and KEGG pathway enrichment analysis showed that neohesperidin's treatment of osteoporosis through PRKACA target mainly depended on biological processes such as protein phosphorylation and protein autophosphorylation,acting on endocrine resistance,proteoglycan in cancer,and estrogen signaling pathway to play a therapeutic role.Molecular docking results showed that neohesperidin had a certain binding ability to the protein corresponding to the target PRKACA.(2)The results of alkaline phosphatase staining showed that neohesperidin could promote the expression of alkaline phosphatase in the early stage of osteogenic differentiation of mesenchymal stem cells.Alizarin red staining showed that neohesperidin could promote the mineralization of osteogenic differentiation of mesenchymal stem cells.RT-qPCR results showed that neohesperidin could increase the mRNA expression of alkaline phosphatase,PRKACA,and osteocalcin.(3)These results indicate that neohesperidin may promote osteogenic differentiation through PRKACA target on the estrogen signaling pathway to prevent and treat osteoporosis.

BACKGROUND:As a degradable scaffold material for bone tissue engineering,lactic acid is widely used in tissue regeneration and repair research,and plays an important role in promoting tissue healing,new bone formation and angiogenesis. OBJECTIVE:To observe the effect of lactic acid degradation products on osteoclasts and to investigate the effects of lactic-interfered osteoclast conditioned medium on the proliferation,migration and tube-forming capacity of human umbilical vein endothelial cells. METHODS:(1)The mouse monocyte macrophage cell line RAW264.7 at logarithmic growth period was selected,and adherent cells were cultured in the osteoclast induction medium(DMEM medium with nuclear factor-κB receptor-activating factor ligand and 10%fetal bovine serum)containing different concentrations of lactic acid(0,5,10,20 mmol/L).After 5 days of culture,tartrate-resistant acid phosphatase staining and cytoskeletal fibrillar actin staining were conducted.After 24 hours of culture,RT-PCR was used to detect the mRNA expression of tartrate-resistant acid phosphatase 5.(2)RAW264.7 cells at logarithmic growth period were selected and adherent cells were divided into two groups.Control group was cultured in the osteoclast induction medium,while experimental group was cultured in the osteoclast induction medium containing 10 mmol/L lactic acid.After 5 days of culture,the medium in each group was removed and the cells in the two groups were cultured in the serum-free DMEM medium for another 24 hours.Cell supernatant was then collected and used as the conditioned medium after mixed with an equal volume of DMEM medium containing 10%fetal bovine serum.Human umbilical vein endothelial cells at the logarithmic growth phase were taken and separately co-cultured with the conditioned medium of the control and experimental groups.The proliferation,migration and tube-forming ability of human umbilical vein endothelial cells were observed by cell counting kit-8 assay,migration assay,scratch assay and tube-forming assay.The mRNA and protein expression of angiogenesis-related genes and proteins were observed by RT-PCR and western blot. RESULTS AND CONCLUSION:Tartrate-resistant acid phosphatase staining and cytoskeletal fibrillar actin staining showed that 5 and 10 mmol/L lactic acid promoted osteoclastic differentiation of RAW264.7 cells and the promoting effect of 10 mmol/L lactate was more significant.RT-PCR results showed that the expression of tartrate-resistant acid phosphatase-5 mRNA of osteoclast-related genes was the highest when the lactic acid concentration was 5,10,and 20 mmol/L(P<0.05),especially 10 mmol/L.Compared with the control group,the proliferation,migration and tube-forming abilities of human umbilical vein endothelial cells were significantly increased in the experimental group(P<0.05).Compared with the control group,the expression levels of vascular endothelial growth factor and angiogenin 1 mRNA and protein were increased in the experimental group(P<0.05).To conclude,lactate-induced osteoclast conditioned medium could promote the angiogenesis of endothelial cells,and the mechanism may be related to the promotion of the expression of vascular endothelial growth factor and angiogenin 1.

BACKGROUND:Traditional Chinese medicine has rich experience and unique advantages in the empirical treatment of phlegm-heat and Fu-organs excess syndrome of ischemic stroke.In order to further explore the therapeutic targets and mechanisms of traditional Chinese medicine for this disease,it is crucial to establish a stable and reliable animal model of phlegm-heat and Fu-organs excess syndrome combined with empirical symptoms of ischemic stroke. OBJECTIVE:To explore the establishment method and evaluation system of the rat model of ischemic stroke with phlegm-heat and Fu-organ excess syndrome. METHODS:Sixty male Sprague-Dawley rats were randomly divided into four groups:blank control group(n=12),ischemic stroke group(n=18),disease+syndrome group(n=18),phlegm-heat and Fu-organ excess syndrome group(n=12),all of which were given high-fat diet for 25 days.On the 26th day,the rats in the blank control group and ischemic stroke group were intragastrically given normal saline and high fat diet,while those in the other two groups were intragastrically given autologous feces suspension and high fat diet for 3 continuous days.After gavage,ischemic stroke models were established using the suture method in the ischemic stroke group and disease+syndrome group.The changes in diet,water intake,body mass,body temperature,fecal traits,nasal secretions,sputum in the throat,and tongue image were recorded.Neurological deficits,tongue image,blood lipid levels,morphological changes of brain tissue and carotid artery,and the serum levels of motilin and somatostatin were detected. RESULTS AND CONCLUSION:Compared with the control group,the rats in the disease+syndrome group had shortness of breath,listlessness,irritability,bradykinesia,a large number of secretions around the nose,audible and heavy sputum in the throat,decreased diet and water intake,increased body mass,body temperature,and slingual vein score,decreased fecal pellet count,Bristol score and fecal moisture content,increased serum total cholesterol,triglyceride,low-density lipoprotein and somatostatin levels,decreased motilin level,increased neurological deficit score,significant pathological changes of the carotid artery,and significant morphological changes of the brain tissue.The ischemic stroke group only showed pathological changes of ischemic brain tissue,without the characteristics of phlegm-heat and Fu-organ excess syndrome.The phlegm-heat and Fu-organ excess syndrome group could present with the typical characteristics of traditional Chinese medicine syndromes,without the pathological changes of brain tissue with ischemic stroke.To conclude,the compound modeling method of high-fat induction combined with suture method and autologous feces gavage can establish an animal model of ischemic stroke with phlegm-heat and Fu-organ excess syndrome.

BACKGROUND:The diabetic microenvironment can cause excessive M1 polarization of macrophages,and this hyperglycemic inflammatory state can inhibit osteogenic differentiation of bone marrow mesenchymal stem cells,thus affecting the healing of diabetic bone defects.Studies have indicated that mogroside V possesses anti-inflammatory,antioxidant,and hypoglycemic properties.However,its potential to modulate M1 polarization of macrophages and osteogenic differentiation of bone marrow mesenchymal stem cells under high glucose and inflammatory condition remains unclear. OBJECTIVE:To explore the effect of mogroside V on regulating M1 macrophage polarization and its effect on osteogenic differentiation of bone marrow mesenchymal stem cells under high glucose and inflammatory condition. METHODS:Murine diabetic models were established using C57BL/6 mice.Bone marrow-derived macrophages were isolated from tibia and fibula of normal and diabetic mice,and cultured in low-glucose and high-glucose media.Then M1 polarization of bone marrow-derived macrophages was induced using lipopolysaccharide and interferon-γ.Bone marrow-derived macrophages were treated with 160,320,and 640 μmol/L mogroside V.Flow cytometry was employed to determine the proportion of F4/80+CD86+cells.qRT-PCR was utilized to assess mRNA expression levels of inducible nitric oxide synthase,interleukin 1β,and interleukin 6.ELISA was employed to evaluate tumor necrosis factor-α secretion in bone marrow-derived macrophage supernatants.Bone marrow mesenchymal stem cells were isolated from tibia and fibula of C57BL/6 suckling mice,and induced osteogenic differentiation using low-or high-glucose osteogenic induction medium.Bone marrow mesenchymal stem cells were treated with M1 macrophage-conditioned mediums with or without 320 μmol/L mogroside V in osteogenic differentiation process.qRT-PCR was employed to assess the mRNA expression of alkaline phosphatase,Runt-related factor 2,osteocalcin,and osteopontin on day 14 after osteogenic induction.Alizarin red staining and quantitative analysis were conducted to evaluate calcium deposition on day 21 after osteogenic induction. RESULTS AND CONCLUSION:(1)Flow cytometry results showed that with the treatment of 320 and 640 μmol/L mogroside V,the proportion of F4/80+CD86+bone marrow-derived macrophages was significantly lower than that in the high-glucose control group(P<0.05).(2)qRT-PCR results showed that with the treatment of 160,320,and 640 μmol/L mogroside V,the mRNA expression levels of inducible nitric oxide synthase and interleukin 6 were significantly lower than that in the high-glucose control group(P<0.05).With the treatment of 320 and 640 μmol/L mogroside V,the mRNA expression level of interleukin 1β was significantly lower than that in the high-glucose control group(P<0.05).(3)ELISA results exhibited that with the treatment of 160,320,and 640 μmol/L mogroside V,the tumor necrosis factor-α secretion level was significantly lower than that in the high-glucose control group(P<0.05).(4)With the treatment of 320 μmol/L mogroside V,calcium salt deposition was increased in bone marrow mesenchymal stem cells under high glucose and inflammatory conditions(P<0.05),and the mRNA relative expression levels of alkaline phosphatase,Runt-related factor 2,and osteopontin were increased(P<0.05).These findings indicate that mogroside V can promote osteogenic differentiation of bone marrow mesenchymal stem cells by inhibiting the M1 polarization of bone marrow-derived macrophages under high glucose and inflammatory conditions and reducing the generation of inflammatory factors.

Objective:To investigate the improvement of radiation-induced cognitive dysfunction and its preliminary mechanism by Ilex pubescens var.kwangsiensis.Methods:SPF grade male Kunming mice were randomly divided into control group(Control),Ilex pubescens var.kwangsiensis(IP),radiation group(Rad)and radiation+Ilex pubescens var.kwangsiensiss group(Rad+IP),with 10 mice in each group.Morris water maze test and dark avoidance test were used to detect the changes in cognitive function of mice before and after drug intervention.Hematoxylin-eosin(HE)staining and electron microscopy were used to observe the changes of brain histopathology and ultrastructure in the hip-pocampus of mice.The expressions of nuclear factor E2-associated factor 2(Nrf2),heme oxygenase 1(HO-1),man-ganese superoxide dismutase(MnSOD),B-cell lymphoma/leukemia-2(Bcl-2)and Bcl-2 associated X protein(Bax)were detected by immunohistochemical assay.Results:The expressions of MnSOD and Bcl-2 in hippocampal tissue of mice in Rad group were significantly decreased,the expressions of Nrf2,Bax and HO-1 were significantly increased,and the nuclear translocation of Nrf2 was also significantly increased.After intervention,he expressions of MnSOD,Nrf2 and HO-1 were significantly increased,and the apoptosis rate was significantly decreased.Conclusion:Ilex pu-bescens var.kwangsiensis improved cognitive dysfunction in mice after radiation,and the mechanism may be related to Nrf2 activation and regulation of HO-1 transcription and expression,reducing oxidative stress damage.

Objective To evaluate the predictive value of functional liver imaging score(FLIS)based on preoperative gadolinium ethoxybenzyl diethylenetriamine pentaacetic acid(Gd-EOB-DTPA)enhanced MRI for post-hepatectomy liver failure(PHLF)in patients with hepatocellular carcinoma(HCC).Methods The data of HCC patients who underwent extensive hepatectomy and preoperative Gd-EOB-DTPA enhanced MRI were analyzed retrospectively.The FLIS was scored based on the three features including liver parenchyma enhancement,biliary excretion and portal vein signal enhancement in hepatobiliary phase images,and the consistency between different observers was evaluated.Logistic regression model and receiver operating characteristic(ROC)curve were used to analyze the ability of FLIS to predict the PHLF.Results PHLF occurred in 29 of 120 HCC patients(24.2%).The intraclass correlation coefficient(ICC)of FLIS evaluated by two observers was 0.944.Multivariate logistic regression analysis showed that FLIS was an independent predictor of PHLF of HCC patients[odds ratio(OR)0.520,95%confidence interval(CI)0.355-0.726;P<0.001].The area under the curve(AUC)of FLIS for predicting the PHLF was 0.709,the optimal diagnostic threshold was 4,and the corresponding sensitivity and specificity were 78.0%and 58.6%.Conclusion Preoperative FLIS can predict the PHLF of HCC patients,which may help to make more accurate treatment plans for HCC patients.

Objective:To explore the mediating role of rumination thinking between demoralization and quality of life in malignant tumor patients, provide guidance and reference for helping tumor patients overcome rumination thinking and demoralization and improve quality of life.Methods:From February 2020 to June 2022, 189 patients with malignant tumors admitted to the Department of Oncology of the First Affiliated Hospital of Guangxi Medical University were selected by convenience sampling method as the research objects, and a cross-sectional survey was conducted using general information questionnaire, Demoralization Scale-Mandarin Version, Ruminative Responses Scale, Punctional Assessment of Cancer Therapy-General.Results:Among 189 malignant tumor patients, there were 102 males, 87 females, aged (43.54 ± 13.12) years old. The total score of loss of demoralization was (34.37 ± 10.34) points, the total score of rumination thinking was (41.01 ± 17.10) points, the total score of quality of life was (48.51 ± 15.41) points. The Pearson analysis results showed that the total score of demoralization in malignant tumor patients was negatively correlated with the total score of quality of life ( r = -0.502, P<0.01); the total score of rumination thinking was negatively correlated with the total score of quality of life ( r = -0.465, P<0.01), and the total score of demoralization was positively correlated with the total score of rumination thinking ( r = 0.628, P<0.01). Bootstrap mediation test results showed that ruminant thinking played a partial mediating effect between demoralization and quality of life of patients with malignant tumors, accounted for 30.9% of the total effect. Conclusions:Rumination plays a partially mediating role in the demoralization and quality of life of patients with malignant tumors, suggesting that clinical staff can improve the quality of life of patients with tumors by developing a systematic and comprehensive cognitive-behavioral intervention strategy to improve the demoralization and rumination.

Objective The aim of this study was to evaluate the efficacy and side effects of sacubitril/valsartan in the treatment of patients with chronic kidney disease(CKD)at stage 5 with resistant hypertension,and to explore the cardiovascular benefits and security of medical in the patients.Methods Patients with CKD5 resistant hypertension diagnosed and treated in the First Affiliated Hospital of Guangxi Medical University from September 2020 to March 2022 were selected and divided into the observation group(treated with routine treatment of kidney disease at end-stage and sacubitril/valsartan)and control group(include droutine treatment of renal disease at end-stage and ACEI or ARB drugs)according to treatment strategy.The patients in both two groups were treated with adequate dialysis treatment and conventional drug treatment of renal disease at end-stage.The patients were followed up for at least 3 months,the clinical efficacy of three months after treated with sacubitril/valsartan was observed,and the efficacy indicators and security indicators and adverse cardiovascular events were observed,the occurrence of adverse effects during the period of drug use were compared with the control group.Results A total of 110 patients were included in this study and there were 55 cases in each group.There were no significant differences in gender,age,age of dialysis,etiology,dialysis mode and blood pressure between the two groups(P>0.05).The Systolic blood pressure(SBP),diastolic blood pressure(DBP),b-type urinary natriuretic peptide precursor(Pro-BNP)and cardiac function grade in the observation group after treatment was significantly decreased compared with before treatment.The left ventricular ejection fraction(LVEF)and the ratio of LVEF<50%in the observation group was significantly reduced after treatment(P<0.05).SBP,DBP and Pro-BNP decreased 3 months after treatment compared with the baseline before treatment,and improved significantly in the first month after treatment(P<0.05).The decrease of DBP and BNP before and after treatment was significantly different between the two groups,and the decrease of DBP and BNP was more significant in the observation group(P<0.05).The difference of LVEF and left ventricular end diastolic diameter(LVEDD)between the two groups before and after treatment was statistically significant,and the improvement was more obvious in the observation group(P<0.05).There were no significant differences in the safety indicators of serum potassium,estimated glomerular filtration rate(eGFR)and liver function between two groups before and after treatment(P>0.05).In terms of adverse reactions,only 1 case in the control group developed hyperkalemia within 3 months of follow-up,and no hypotension or other adverse reactions occurred in the two groups.Conclusions The treatment of patients with CKD stage 5 hypertension with sacubitril/valsartan has obvious cardiovascular benefits.Sacubitril/Valsartan has efficacy in lowering blood pressure,improving cardiac function and reducing volume load,with less adverse events and higher safety than control group.

Objective The aim of this study was to screen and verify the proteins interacting with Vimentin,investigate the regulatory relationship between FABP5 and candidate proteins,and further explore the mechanism of FABP5 in hepatocellular carcinoma.Methods Immunoprecipitation combined with tandem mass spectrometry(IP-MS)was used to screen the proteins that bind to FABP5.The binding relationship between FABP5 and candi-date interacting proteins was verified from the exogenous and endogenous levels by Co-immune precipitation assay(Co-IP).RT-qPCR,Western blot and immunofluorescence were used to observe the effect of knockdown FABP5 on the transcription and translation of Vimentin in HCC cells.The effect of overexpressing FABP5 on the cytoskeleton of HCC cell was observed by phalloidin staining.Results 336 potential target proteins that bind to FABP5 were identi-fied through IP-MS.Based on literature,five candidate proteins related to tumors were selected,namely PRDX1,PRSS3,PKM,HSP90AA1,and Vimentin.The binding relationship between FABP5 and Vimentin protein was con-firmed through both exogenous and endogenous Co-IP.Knockdown FABP5 has no significant effect on the expression of Vimentin mRNA,but it can inhibit the expression of Vimentin protein,and overexpression of FABP5 can affect the cytoskeleton of HCC cell.Conclusions FABP5 promotes the migration and invasion of HCC cells by the regula-tion of Vimentin and the influence of cytoskeletal remodeling,and thus it is expected to be a potential target for anti-HCC and provide new ideas for the treatment of HCC.

Objective To explore the association between serum trace element levels in early pregnancy and gestational dia-betes mellitus(GDM),and the mediating effect of bile acid metabolism changes in this association.Methods A nested case-con-trol study was designed based on the Guangxi Zhuang Birth Cohort.A total of 248 pregnant women(case group=124,control group=124)were included from June 2015 to July 2019 in Nanning city.The concentrations of 8 trace elements and 31 bile acids in serum were measured in early pregnancy.Conditional logistic regression and BKMR models were used to analyze the associa-tion and combined effect between trace elements and GDM risk,respectively.Orthogonal partial least squares-discriminant anal-ysis(OPLS-DA)was used to screen potential bile acid biomarkers associated with GDM,and then conditional logistic regression was used to determine the association between specific bile acid levels and GDM risk.Multiple linear regression was used to e-valuate the association of serum trace element concentrations with differential bile acid metabolites.Mediation analysis was used to evaluate the mediating role of bile acids in the relationship between trace element exposure and GDM.Results After adjus-ting for confounding factors,serum vanadium(V)was found to be positively associated with the risk of GDM,while chromium(Cr),manganese(Mn),zinc(Zn),selenium(Se)and molybdenum(Mo)were negatively correlated with the risk of GDM(all P＜0.05).The OPLS-DA model and conditional Logistic regression analysis showed that taurocholic acid(TCA),glycochenodeoxy-cholic acid 3-sulfate(GCDCA-3S),glycochenodeoxycholic acid-3-O-β-glucuronide(GCDCA-3Gln),glycoursodeoxycholic acid-3-sulfate(GUDCA-3S),taurodeoxycholic acid-3-sulfate(TDCA-3S),and chenodeoxycholic acid(CDCA)might be potential bile acid metabolic markers of GDM(all P＜0.05).The concentrations of multiple trace elements were also significantly correlated with the levels of specific bile acids(all P＜0.05).Mediation analysis showed that GCDCA-3Gln and TCA mediated the associa-tion between serum Zn and Se and GDM risk,respectively(all P＜0.05).Conclusion Serum trace elements such as V and Cr are significantly associated with the risk of GDM in early pregnancy,and changes in bile acid metabolism may precede the occur-rence of GDM.It is suggested that the effect of trace elements on the metabolism of bile acids,especially conjugated bile acids,may be one of the mechanisms affecting the risk of GDM.