Objective To investigate the effects of aloperine(ALO)on bone metabolism in osteoporosis(OP)mice via Wnt/β-catenin signaling pathway-mediated autophagy and apoptosis.Methods Sixty mice were randomly allocated into six groups(n=10 per group):Sham group(sham-operated mice),OP group(osteoporosis model induced by bilateral ovariectomy),L-ALO group(OP mice intraperitoneally injected with 10 mg/kg aloper-ine),M-ALO group(OP mice intraperitoneally injected with 20 mg/kg aloperine),H-ALO group(OP mice intra-peritoneally injected with 30 mg/kg aloperine),and EV group(OP mice administered 0.09 mg/kg estradiol valerate).Bone mineral density and microstructure of the tibia were assessed.Hematoxylin and eosin(HE)staining was per-formed to examine the morphology of tibial bone tissue.Serum levels of OCN,OPG,ALP,Ca,and P were measured using ELISA.Protein expression levels of LC3-Ⅱ,LC3-Ⅰ,Beclin-1,P62,Caspase-3,Caspase-9,Bax,Wnt3a,β-catenin,and C-Myc were analyzed by Western blot.Autophagosomes were visualized using immunofluorescence.Results Compared with the Sham group,the bone mineral density(BMD)and trabecular thickness in the OP group were significantly reduced,while trabecular separation,bone surface area,and volume were significantly increased(P<0.05).The levels of OPG,OCN,Ca,and P were significantly downregulated,whereas ALP levels were significantly upregulated(P<0.05).Additionally,the LC3-Ⅱ/LC3-Ⅰ ratio and expression levels of Beclin-1,Wnt3a,β-catenin,and C-Myc proteins were significantly decreased,while the expression levels of P62,Caspase-3,Caspase-9,and Bax proteins were significantly increased(P<0.05).Compared with the OP group,the L-ALO,M-ALO,and H-ALO groups exhibited significant increases in BMD and trabecular thickness,along with significant decreases in trabecular separation,bone surface area,and volume(P<0.05).The levels of OPG,OCN,Ca,and P were significantly upregulated,while ALP levels were significantly downregulated(P<0.05).Furthermore,the LC3-Ⅱ/LC3-Ⅰ ratio and expression levels of Beclin-1,Wnt3a,β-catenin,and C-Myc proteins were significantly increased,while the expression levels of P62,Caspase-3,Caspase-9,and Bax proteins were significantly decreased(P<0.05).In contrast,compared with the OP group,the EV group showed significant increases in BMD and trabecular thickness,as well as significant decreases in trabecular separation,bone surface area to volume ratio(P<0.05).The levels of OPG,OCN,Ca,and P were significantly upregulated,while ALP levels were significantly downregulated(P<0.05).Moreover,the LC3-Ⅱ/LC3-Ⅰ ratio and expression levels of Beclin-1,Wnt3a,β-catenin,and C-Myc proteins were significantly increased,while the expression levels of P62,Caspase-3,Caspase-9,and Bax proteins were significantly decreased(P<0.05).Conclusions Peanine may promote autophagy in osteoblasts and inhibit their apoptosis,thereby improving bone metabolism in OP mice.This effect may be mediated through the activation of the Wnt/β-catenin signaling pathway.

Objective To investigate the impact of serine/threonine-protein kinase 1(AKT1)-mediated autophagy in hepatocellular carcinoma(HCC)cells on their sensitivity to 125I seed irradiation.Methods ① iProX database and STRING12.0 website were utilized to analyze the proteomic data of HCC before and after 125I seed irradiation to explore the differentially expressed proteins and associated functional connections.Meanwhile,The Cancer Genomics Atlas(TCGA)database was employed to analyze the relationship between AKT1 expression level and the survival of HCC patients.② Human HCC cell lines HUH7 and Hep3B were exposed to continuous irradiation from 125I radioactive seeds with an initial apparent activity of 0.8 mCi per seed for approximately 120 h,accumulating a total dose of 8 Gy,while the control cells were cultured under normal condition for 120 h.③ Autophagy inhibitor,chloroquine(CQ)and inducer,rapamycin(RAPA)were used to treat the HCC cells respectively to establish the CQ group and the RAPA group.The lentiviral transfection technique was employed to construct the HCC cells with overexpressed AKT1,namely the AKT1 group.The HCC cells treated in the same way were continuously irradiated with 125I seeds for 120 h to construct the CQ+125I group,the RAPA+125I group,and the AKT1+125I group.④The changes in microtubule-associated protein light chain 3(LC3),p62,AKT1 and p-AKT1 were detected by Western blotting.Cell immunofluorescence assay was employed to observe the expression of autophagy related proteins,such as LC3.The colony forming ability and apoptotic rate were detected with plate cloning assay and flow cytometry.Results ① Continuous irradiation with 125I seeds resulted in decreased expression of p62 and increased ratio of LC3Ⅱ/LC3Ⅰ(P＜0.05)when compared with the negative control(NC)group.Immunofluorescence assay revealed more green fluorescence spots of LC3.When compared with the 125I group,the CQ+125I group had significantly increased expression of p62(P＜0.01),decreased ratio of LC3Ⅱ/LC3Ⅰ(P＜0.01),lower apoptotic rate(P＜0.01),and more colony formations(P＜0.01).In contrast,the results in the RAPA+125I group were opposite to those of the CQ+125I group.② Analysis on the iProX database showed that the expression of AKT1 was decreased in the irradiated group,and analysis on the TCGA database indicated that high expression of AKT1 predicted a poor prognosis for HCC patients(P＜0.01).③After irradiation with 125I seeds,the expression of AKT1 at both the RNA and protein levels was decreased in the 125I group(P＜0.01).After overexpression of AKT1,the level of autophagy was decreased(P＜0.05).Irradiation of HCC cells with overexpressed AKT1 using 125I seeds could partially restore the level of autophagy.In the AKT1+125I group,the expression of AKT1,pAKT1 and p62 were all decreased,and the ratio of LC3Ⅱ/LC3Ⅰwas increased than the AKT1 group(P＜0.05).④ The apoptotic rate of the AKT1+125I group was lower than that of the 125I group(P＜0.05)and higher than that in the AKT1 group(P＜0.05).In HUH7 cells,the clonogenic ability of the AKT1+125I group was higher than that of the 125I group(P＜0.05).In Hep3B cells,the clonogenic ability of the AKT1+125I group was higher than that of the 125I group,and the clonogenic ability of the AKT1 group was higher than that of the NC group(P＜0.05).Conclusion 125I seed irradiation induce lethal autophagy in HCC cells by reducing the expression of AKT1,providing a new theoretical basis for the implantation of 125I radioactive seeds in the treatment of HCC.

Objective To investigate the effects of aloperine(ALO)on bone metabolism in osteoporosis(OP)mice via Wnt/β-catenin signaling pathway-mediated autophagy and apoptosis.Methods Sixty mice were randomly allocated into six groups(n=10 per group):Sham group(sham-operated mice),OP group(osteoporosis model induced by bilateral ovariectomy),L-ALO group(OP mice intraperitoneally injected with 10 mg/kg aloper-ine),M-ALO group(OP mice intraperitoneally injected with 20 mg/kg aloperine),H-ALO group(OP mice intra-peritoneally injected with 30 mg/kg aloperine),and EV group(OP mice administered 0.09 mg/kg estradiol valerate).Bone mineral density and microstructure of the tibia were assessed.Hematoxylin and eosin(HE)staining was per-formed to examine the morphology of tibial bone tissue.Serum levels of OCN,OPG,ALP,Ca,and P were measured using ELISA.Protein expression levels of LC3-Ⅱ,LC3-Ⅰ,Beclin-1,P62,Caspase-3,Caspase-9,Bax,Wnt3a,β-catenin,and C-Myc were analyzed by Western blot.Autophagosomes were visualized using immunofluorescence.Results Compared with the Sham group,the bone mineral density(BMD)and trabecular thickness in the OP group were significantly reduced,while trabecular separation,bone surface area,and volume were significantly increased(P<0.05).The levels of OPG,OCN,Ca,and P were significantly downregulated,whereas ALP levels were significantly upregulated(P<0.05).Additionally,the LC3-Ⅱ/LC3-Ⅰ ratio and expression levels of Beclin-1,Wnt3a,β-catenin,and C-Myc proteins were significantly decreased,while the expression levels of P62,Caspase-3,Caspase-9,and Bax proteins were significantly increased(P<0.05).Compared with the OP group,the L-ALO,M-ALO,and H-ALO groups exhibited significant increases in BMD and trabecular thickness,along with significant decreases in trabecular separation,bone surface area,and volume(P<0.05).The levels of OPG,OCN,Ca,and P were significantly upregulated,while ALP levels were significantly downregulated(P<0.05).Furthermore,the LC3-Ⅱ/LC3-Ⅰ ratio and expression levels of Beclin-1,Wnt3a,β-catenin,and C-Myc proteins were significantly increased,while the expression levels of P62,Caspase-3,Caspase-9,and Bax proteins were significantly decreased(P<0.05).In contrast,compared with the OP group,the EV group showed significant increases in BMD and trabecular thickness,as well as significant decreases in trabecular separation,bone surface area to volume ratio(P<0.05).The levels of OPG,OCN,Ca,and P were significantly upregulated,while ALP levels were significantly downregulated(P<0.05).Moreover,the LC3-Ⅱ/LC3-Ⅰ ratio and expression levels of Beclin-1,Wnt3a,β-catenin,and C-Myc proteins were significantly increased,while the expression levels of P62,Caspase-3,Caspase-9,and Bax proteins were significantly decreased(P<0.05).Conclusions Peanine may promote autophagy in osteoblasts and inhibit their apoptosis,thereby improving bone metabolism in OP mice.This effect may be mediated through the activation of the Wnt/β-catenin signaling pathway.

BACKGROUND: Men who have sex with men (MSM) have become a high risk population of HIV infection due to their risky sexual behaviors. The latent pattern of psychosocial characteristics plays an important effect in HIV-related risky behaviors among HIV-negative MSM. METHOD: Participants were recruited from Wuhan, Nanchang, and Changsha city from September 2017 to January 2018. Social support was assessed by the multidimensional scale of social support, Connor-Davidson Resilience scale-10 items for reliance, the assessment of Stigma towards Homosexuality for sexual minority stigma, the Likert subscale of nondisclosure for identity concealment, the ACE questionnaire-Kaiser-CDC for adverse childhood experience, the Centers for Epidemiological Studies Depression Scale for depression. Latent profile analysis (LPA) and multivariate regression were used to analyze the data. RESULTS: Three psychosocial characteristic patterns were revealed by the LPA. "Social support and resilience group" (SR group), "Identity concealment group" (IC group) and "Adverse childhood experience" (ACE group) were identified, respectively. In comparison with "SR group", "IC group" have a higher likelihood of one-night male partners (AOR = 2.74, 95%CI = [1.54, 4.90]), both fixed and one-night male partners (AOR = 2.01, 95%CI = [1.34, 3.01]) and HIV-unsure male partner (AOR = 2.12, 95%CI = [1.44, 3.13]). Similarly, "ACE group" were more likely having inconsistent condom use (AOR = 2.58, 95%CI = [1.41, 4.73]), and having sex with HIV-positive male partner (AOR = 4.90, 95%CI = [1.95, 12.30]) with comparison of "SR group". In addition, we further revealed that "ACE group" had a higher ratio (90.0%) of inconsistent condom use among MSM whose male partners were HIV-positive. CONCLUSIONS Six important psychosocial factors were divided into three latent pattern classes. Compared with "SR group", "IC group" and "ACE group" were more likely to engage in HIV-related risky sexual behaviors. Further research may pay more attention to "IC group" and "ACE group" for targeted intervention.
Humans ; Male ; HIV Infections/epidemiology* ; Homosexuality, Male/psychology* ; Risk Factors ; Sexual and Gender Minorities/psychology* ; Sexual Behavior/psychology*

Objective To investigate the association of serum ghrelin level with cognition, hippocampal volume, and proton magnetic resonance spectrum ( [ 1 H ]-MRS ) in patients with type 2 diabetes mellitus ( T2DM) . Methods The T2DM patients cared at the Wuhan Fourth Hospital were recruited. Data on demographic characteristics and clinical parameters were collected. Ghrelin was measured by ELISA assay. Cognitive performance was assessed by the Montreal Cognitive Assessment ( MoCA ) and Repeatable Battery for the Assessment of Neuropsychological Status (RBANS). The changes of metabolites in hippocampus were detected by [1 H]-MRS, including N-acetyl asparate ( NAA) , choline ( Cho) , myo-inositol ( MI) , creatine ( Cr) . All patients were divided into 2 groups[cognitive impairment (CI) and non-cognitive impairment (NCI) groups] by MoCA. Results (1) Compared with patients in NCI group, the serum ghrelin level, bilateral hippocampal volume, and NAA/Cr ratio of [1H]-MRS metabolites in CI group were decreased, but MI/Cr and Cho/Cr ratios were increased(all P<0.05). (2) Serum ghrelin was positively correlated with a variety of RBANS scores ( including immediate memory, attention, delayed memory, and total scores) , bilateral hippocampal volume, and NAA/Cr ratio of [ 1 H]-MRS metabolites in T2DM patients, but it was negatively correlated with MI/Cr ratio and Cho/Cr ratio ( all P<0. 05 ) . ( 3 ) Logistic regression analysis showed that ghrelin was a protective factor of cognition in T2DM patients. Conclusions T2DM patients with cognitive impairment had lower levels of ghrelin, and serum ghrelin was postively correlated with their cognitive performance, hippocampal volume, and [1 H]-MRS metabolites, suggesting that serum ghrelin may be involved in the occurrence and development of cognitive dysfunction in patients with T2DM.

Objective To systematically evaluate the relationship between atherosclerotic renal artery stenosis (ARAS) and cor‐onary artery disease (CAD) and peripheral arterial disease (PAD) .Methods We gathered all case‐control studies about the correla‐tion of ARAS with CAD and PAD in the following databases:Cochrane library ,PubMed ,EMBASE ,Web of science until April , 2014 .Two reviewers extracted all relevant datas from the screened documents independently according to exclusion and inclusion criteria ,RevMan 5 .2 software were used to conduct Meta‐analysis .Results Fourteen trials were included .Meta‐analysis showed that :the OR (95% CI)of CAD with 1 vascular lesions ,2 vascular lesions ,3 vascular lesions and left main stenosis ,PAD and ARAS were 0 .70(0 .59-0 .82) ,1 .28(1 .10 -1 .48) ,2 .09(1 .69 -2 .59) ,1 .82(1 .40 -2 .36) ,3 .68(2 .21 -6 .10) with statistical signifi‐cance (P<0 .05) .Conclusion CAD with 2 vascular lesions ,3 vascular lesions and left main stenosis ,PAD were connected with ARAS ,CAD with 1 vascular lesions has little relationship with ARAS .