Objective:To investigate the effect of the stress-induced protein sestrin 3(SESN3)on tumor proliferation and the sensitivity to endocrine therapy in hormone receptor-positive(HR+)breast cancer(BC)and its mechanism.Methods:The cancer genome atlas(TCGA)database was used to analyze the expression level of SESN3 in breast cancer tissue and paracancerous tissue.The clinical samples of breast cancer tissue and paracancerous tissue were collected,and immunohistochemistry(IHC)microarray was used to vali-date the expression of SESN3.Western blot was used to measure the protein expression level of SESN3 in BC cell lines with different pathological subtypes.HR+BC cell lines were used to establish the models of SESN3 overexpression and knockdown,and cell counting kit-8(CCK-8)assay and colony formation assay were used to assess the proliferative capacity of cells;Western blot was used to ana-lyze cell cycle alterations;a subcutaneous xenograft mouse model was used to validate the effect of SESN3 on proliferation in vivo.The bioinformatics analysis and Western blot were used to investigate the downstream mechanisms of SESN3 in regulating proliferation.Results:The analysis of TCGA database and clinical samples showed that the expression of the SESN3 gene in BC tissue was signifi-cantly lower than that in paracancerous tissue,and the patients with high SESN3 expression had a longer progression-free survival time.Compared with the other subtypes of BC cell lines,the HR+BC cells lines MCF-7 and T-47D had a relatively high expression level of SESN3.MCF-7 and T-47D cell lines were used to establish the cell models of knockdown(MCF-7KD and T-47DKD)and overexpres-sion(MCF-7OE and T-47DOE),and compared with the control group,MCF-7KD and T-47DKD cells showed increases in proliferative capacity and the proportion of cells arrested in the G2 and M phases.In vivo tumorigenesis experiments showed a significant increase in tumor growth in mice in the MCF-7KD group.The bioinfor-matics analyses showed a significant negative correlation between the mammalian Target of Rapamycin(mTOR)signaling pathway and SESN3 expression,and Western blot validation showed a significant increase in the expression of mTOR in MCF-7KD and T-47DKD cells,as well as reductions in the protein expression levels of autophagy-related genes.Subsequently,we further confirmed that the knockdown of MCF-7 and T-47D cells led to a decreased sensitivity to endocrine therapy.In addition,MCF-7、T-47D、MCF-7KD、and T-47DKD cell lines were treated with the mTOR inhibitor everolimus(EVE),and there was a significant reduction in proliferative capac-ity in the MCF-7KD group and the T-47DKD group;since the mTOR pathway reduced the sensitivity to endocrine therapy by reducing cell apopto-sis,there was a significant increase in the sensitivity to endocrine therapy with tamoxifen(TAM).Conclusion:The inte-grated database analysis,clinical sample validation,and in vivo/in vitro experimental models show that SESN3 inhibits the proliferation of HR+BC cells via the mTOR pathway and enhances the sensitivity to endocrine therapy.

Objective To investigate the effect of solasonine,an active component of Solanum nigrum,on proliferation and apoptosis of non-small cell lung cancer PC9 cells.Methods PC9 cells were treated with 2,5,10,15,20,or 25 μmol/L solasonine,and the changes in cell proliferation were examined using CCK-8 assay.Tetramethyl rhodamine ethyl ester(TMRE)was used to detect the changes in mitochondrial membrane potential,and caspase-3/7 detection kit and GreenNuc? caspase-3/Annexin V-mCherry kit for live cell were used to analyze the changes in caspase-3 of the cells.Annexin V-FITC/PI double staining was employed to analyze the apoptosis rate of the cells.The effect of PTEN inhibitors on solasonine-induced cell apoptosis was examined by detecting apoptosis-related protein expressions using Western blotting.Results Solasonine treatment for 24,48,and 72 h significantly lowered the viability of PC9 cells.The cells treated with solasonine for 24 h showed significantly decreased mitochondrial membrane potential and increased cell apoptosis with enhanced caspase-3/7 and caspase-3 activities and expression of cleaved caspase-3.Solasonine treatment significantly decreased phosphorylation levels of PI3K and Akt,increased the protein expressions of PTEN and Bax,and lowered the expression of Bcl-2 protein in the cells.Conclusion Solasonine inhibits proliferation and induces apoptosis of PC9 cells by regulating the Bcl-2/Bax/caspase-3 pathway and its upstream proteins.

Objective To investigate the effect of solasonine,an active component of Solanum nigrum,on proliferation and apoptosis of non-small cell lung cancer PC9 cells.Methods PC9 cells were treated with 2,5,10,15,20,or 25 μmol/L solasonine,and the changes in cell proliferation were examined using CCK-8 assay.Tetramethyl rhodamine ethyl ester(TMRE)was used to detect the changes in mitochondrial membrane potential,and caspase-3/7 detection kit and GreenNuc? caspase-3/Annexin V-mCherry kit for live cell were used to analyze the changes in caspase-3 of the cells.Annexin V-FITC/PI double staining was employed to analyze the apoptosis rate of the cells.The effect of PTEN inhibitors on solasonine-induced cell apoptosis was examined by detecting apoptosis-related protein expressions using Western blotting.Results Solasonine treatment for 24,48,and 72 h significantly lowered the viability of PC9 cells.The cells treated with solasonine for 24 h showed significantly decreased mitochondrial membrane potential and increased cell apoptosis with enhanced caspase-3/7 and caspase-3 activities and expression of cleaved caspase-3.Solasonine treatment significantly decreased phosphorylation levels of PI3K and Akt,increased the protein expressions of PTEN and Bax,and lowered the expression of Bcl-2 protein in the cells.Conclusion Solasonine inhibits proliferation and induces apoptosis of PC9 cells by regulating the Bcl-2/Bax/caspase-3 pathway and its upstream proteins.

Brown adipose tissue (BAT) holds great promise for the prevention and treatment of metabolism diseases through thermoregulation. Polycystic ovary syndrome (PCOS) is a complex condition with anovulation, hyperandrogenism, and polycystic ovaries, and also manifests glucolipid metabolic disorders. Recent researches have shown that transplantation of BAT into a PCOS rat could significantly alleviate the phenotypes. This article reviews the role of BAT in pathogenesis of PCOS, which may provide information for prevention and treatment of PCOS.

Objective To investigate the effect of triggering receptor expression in myeloid cells-1 (TREM-1) on intestinal macrophage apoptosis in rat.Methods In vitro,the achieved rat intestinal macrophages were divided into 3 groups:control group,LPS (Lipopolysaccharides) group and LPS + LP17 group (n =6 holes of culture plate in each).The concentrations of LPS and LP17 were 1 mg/L and 0.1 mg/L,respectively.The intestinal macrophage apoptosis was measured by using TUNEL kit and flow cytometry after culture for 6 h.All data were statistically analyzed using SPSS 18.0 software.Results The shape and growth of rat intestinal macrophages were quite favorable after culture.The membrane marker of intestinal macrophages,CD14 was clearly observed under immunofluorescence.After macrophage was treated with specific procedure,the cell apoptosis found in LPS group (44.33 ± 7.74)％ was significantly higher than that in control group (19.17 ± 6.01) ％ (P=0.000) measured by TUNEL;the cell apoptosis in LPS +LP17 group (28.33 ± 6.53)％ apparently reduced compared with LPS group (44.33 ±7.74) ％ (P =0.004);there was no significant difference in cell apoptosis between control group (19.17 ± 6.01) ％ and LPS + LP17 group (28.33 ± 6.53) ％ (P =0.050).By flow cytometry,the apoptotic cells in LPS group (16.47 ± 1.66) ％ was significantly increased compared with control group (7.70 ± 1.52) ％ (P =0.000);apoptotic cells in LPS + LP17 group (11.47 ± 3.12) ％ was significantly reduced in comparison with LPS group (16.47 ± 1.66) ％ (P =0.018).There was no significant difference in apoptotic cells between control group (7.70±1.52)％ and LPS + LP17 group (11.47±3.12) ％ (P =0.061).Conclusion LP17 can inhibit TREM-1 expression in intestinal macrophages and reduce intestinal macrophage apoptosis.

Objective To investigate the macrophages (Mφ) phenotype mechanism in acute kidney injury caused by severe acute pancreatitis (SAP).Methods Sixty-four male Wistar rats were randomly divided into control group (SO) and SAP group (n =32 in each group).SAP rat model was made by retrograde cholangiopancreatic injection of 5％ sodium taurocholate.At 2,6,12 and 24 h after modeling,the samples of blood and kidney tissue were collected.The levels of blood urea nitrogen (BUN) and creatinine (Cr) were detected by using automatic biochemical analyzer.The expressions of IL-12,TNF-α,IL-10 and TGF-β mRNA of kidney tissue were detected by fluorescence quantitative polymerase chain reaction (QRT-PCR).The levels of CD68,iNOS and Arg-1 were measured by Western blot.Results In the SAP group at each interval,BUN and Cr concentrations were significantly higher than those of the control group (P ＜ 0.01,P ＜ 0.05) ; Compared with the control group,the expressions of IL-12,TNF-α,IL-10 and TGF-β mRNA in renal tissue of SAP group were significantly higher (P ＜ 0.01,P ＜ 0.05).In the SAP group,the levels of CD68,iNOS and Arg-1 were higher than those in the control group.Conclusions Inflammation and inflammatory imbalances may be pathological factors of acute kidney injury following SAP.