Objective:To investigate the effects of Juglone on the apoptosis of osteosarcoma(OS)cells(U2OS and MG63 cells)through the cysteinyl aspartate specific proteinase-3(Caspase-3)/gasdermin E(GSDME)-mediated pyroptosis pathway.Methods:The U2OS and MG63 cells were cultured in vitro and divided into control group,different concentrations(5,10 and 20 μmol·L-1)of Juglone groups and Caspase-3 inhibitor Z-DEVD-FMK group(10 μmol·L-1 Juglone+30 μmol·L-1 Z-DEVD-FMK).The survival rates of cells in various groups were assessed by cell counting kit-8(CCK-8)assay,and the apoptotic rates were detected by flow cytometry.Lactate dehydrogenase(LDH)release assay was used to measure the release rates of LDH from the cells.Western blotting method was used to detect the expression levels of apoptosis-related proteins including B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),cleaved-Caspase-3 and poly(ADP-ribose)polymerase(PARP)and pyroptosis-related proteins including GSDME full form(GSDME-F)and GSDME N-terminal(GSDME-N).The levels of interleukin-1β(IL-1β)and interleukin-18(IL-18)in the cell supernataut in various groups were measured by enzyme-linked immunosorbent assay(ELISA)method.Results:Compared with control group,the survival rates of cells in 5,10,and 20 μmol·L-1Juglone groups were significantly decreased(P＜0.05 or P＜0.01),and the 50％inhibitory concentration(IC50)values of U2OS cells and MG63 cells were 8.4 and 10.2 μmol·L-1,respectively.Compared with control group,the apoptotic rates and LDH release rates of U2OS and MG63 cells in 5 and 10 μmol·L-1Juglone groups were significantly increased(P＜0.05 or P＜0.01).Compared with control group,the expression levels of Bax,cleaved-Caspase-3,and cleaved-PARP proteins in 5 and 10 μmol·L-1 Juglone groups were significantly increased(P＜0.01),while the expression levels of Bcl-2 protein were significantly decreased(P＜0.01).Compared with control group,the levels of IL-1β and IL-18 in cell supernatant in 5 and 10 μmol·L-1Juglone groups were increased(P＜0.01).Compared with control group,the expression levels of cleaved-Caspase-3 and GSDME-N proteins in 5 and 10 μmol·L-1 Juglone groups were significantly increased(P＜0.01),while there was no difference in the expression level of GSDME-F protein(P＞0.05).Compared with 10 μmol·L-1 Juglone group,the expression levels of cleaved-Caspase-3 and GSDME-N in Z-DEVD-FMK group were significantly decreased(P＜0.01),while there was no difference in the expression level of GSDME-F protein(P＞0.05).Conclusion:Juglone can induce the apoptosis of U2OS and MG63 cells and cause the Caspase-3/GSDME-mediated pyroptosis.

Objective To investigate the therapeutic effects of bone marrow mesenchymal stem cells (BMSCs) for radiation-induced lung injury (RILI) and the underlying mechanism. Methods Forty-five healthy adult male C57BL/6 mice were randomly divided into control, model, and BMSCs groups. The model and BMSCs groups received a single irradiation dose of 20 Gy to the chest, while the control group did not receive X-ray irradiation. For the BMSCs group, an injection of 1 × 10⁶ BMSCs cells was administered via the tail vein within 6 h after irradiation. In the 5th week, the lung tissue was taken to observe pathological changes with HE staining; examine the expression of the inflammatory factors interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) with immunohistochemical staining; observe the polarization of macrophages with immunofluorescence staining; and measure the expression of the epithelial-mesenchymal transition markers E-cadherin, N-cadherin, and vimentin proteins by Western blot. Results After radiation, the model group developed pulmonary vasodilation and congestion with septal thickening and inflammatory cell infiltration, and these changes were markedly reduced in the BMSCs group. The model group showed significantly down-regulated expression of IL-6 and TNF-α compared with significantly increased levels in the model group (P < 0.01, P < 0.05). Treatment with BMSCs significantly increased the polarization of lung macrophages towards the M2 type, while significantly decreasing the abnormally increased N-cadherin and vimentin levels in RILI mice (P < 0.05, P < 0.01). Conclusion BMSCs have therapeutic effects for RILI mice, which may be through promoting macrophage polarization from M1 to M2.

Objective:To discuss the effect of curcumin on the proliferation,migration,and invasion of the human prostate cancer C4-2 and LNCaP cells,and to clarify its possible mechanism.Methods:The lentiviral transfection system was used to transfect the C4-2 and LNCaP cells,regarded as shCD147-C4-2 group and shCD147-LNCaP group.RNA interference technology was used to prepare the CD147-silenced cells;the cells transfected with an empty vector were regarded as negative control and divided into shNC-C4-2 group(shNC-C4-2 cells)and shNC-LNCaP group(shNC-LNCaP cells).The C4-2 and LNCaP cells at logarithmic growth phase,as well as shCD147-C4-2 and shCD147-LNCaP cells,were treated with 20 μmol·L-1 curcumin.The morphology of the cells in various groups was observed under microscope at 0 and 24 h of treatment;MTT method was used to detect the proliferation activities of the cells in various groups;cell scratch assay was used to detect the migration rates of the cells in various groups;Western blotting method was used to detect the expression levels of apoptosis,invasion,and migration-related proteins in the cells in various groups.Results:Compared with C4-2 group,the expression of CD147 protein in the cells in shCD147-C4-2 group was significantly decreased after CD147 gene silenting.Compared with LNCaP group,the expression level of CD147 protein in the cells in shCD147-LNCaP group was significantly decreased after CD147 gene silenting.Compared with 0 h of treatment,some cells in C4-2 and LNCaP groups after 24 h of treatment with 20 μmol·L-1 curcumin,showed apoptosis signs with the presence of typical apoptotic bodies.The apoptotic phenomena in shCD147-C4-2 and shCD147-LNCaP groups was reduced.The MTT assay results showed that compared with C4-2+0 μmol·L-1 curcumin group,the proliferation activities of the cells in C4-2+20 μmol·L-1 curcumin group,C4-2+40 μmol·L-1 curcumin group,C4-2+60 μmol·L-1 curcumin group,and C4-2+80 μmol·L-1 curcumin group were decreased(P<0.01).Compared with LNCaP+0 μmol·L-1 curcumin group,the proliferation activity of the cells in LNCaP+20 μ mol·L-1 curcumin group,LNCaP+40 μmol·L-1 curcumin group,LNCaP+60 μmol·L-1 curcumin group,and LNCaP+80 μmol·L-1 curcumin group were decreased(P<0.01).Compared with shNC-C4-2 group,the proliferation activity of the cells in shNC-C4-2+20 μmol·L-1 curcumin group was decreased(P<0.01).Compared with shNC-C4-2+20 μmol·L-1 curcumin group,the proliferation activity of the cells in shCD147-C4-2+20 μmol·L-1 curcumin group was increased(P<0.01).Compared with shNC-LNCaP group,the proliferation activity of the cells in shNC-LNCaP+20 μmol·L-1 curcumin group was decreased(P<0.01);compared with shNC-LNCaP+20 μmol·L-1 curcumin group,the proliferation activity of the cells in shCD147-LNCaP+20 μmol·L-1 curcumin group was significantly increased(P<0.01).The cell scratch healing assay results showed that compared with C4-2 group,the migration rates of the cells in C4-2+20 μmol·L-1 curcumin group and C4-2+40 μmol·L-1 curcumin group after 24 h of treatment were decreased(P<0.01);compared with LNCaP group,the migration rates of the cells in LNCaP+20 μmol·L-1 curcumin group and LNCaP+40 μmol·L-1 curcumin group were increased(P<0.01);compared with shNC-C4-2 group,the migration rate of the cells in shNC-C4-2+20 μmol·L-1 curcumin group was decreased(P<0.01);compared with shNC-C4-2+20 μmol·L-1 curcumin group,the migration rate of the cells in shCD147-C4-2+20 μmol·L-1 curcumin group was significantly increased(P<0.05);compared with shNC-LNCaP group,the migration rate of the cells in shNC-LNCaP+20 μmol·L-1 curcumin group was decreased(P<0.01);compared with shNC-LNCaP+20 μmol·L-1 curcumin group,the garation rate of the cells in shCD147-LNCaP+20 μmol·L-1 curcumin group was significantly increased(P<0.05).The Western blotting results showed that compared with C4-2 group,the expression levels of Bcl-2-associated X protein(Bax),cleaved Caspase-3,and poly ADP-ribose polymerase 1(PARP1)proteins in the cells in C4-2+20 μmol·L-1 curcumin group and C4-2+40 μmol·L-1 curcumin group were significantly increased(P<0.01),and the expression levels of Bcl-2 protein was significantly decreased(P<0.05 or P<0.01);compared with LNCaP group,the expression levels of Bax,cleaved Caspase-3,and PARP1 proteins in the cells in LNCaP+20 μmol·L-1 curcumin group and LNCaP+40 μmol·L-1 curcumin group were significantly increased(P<0.01),and the expression level of Bcl-2 protein in the cells in LNCaP+40 μmol·L-1 curcumin group was decreased(P<0.01);compared with shNC-C4-2 group,the expression levels of Bax,cleaved Caspase-3,and PARP1 proteins in the cells in shNC-C4-2+20 μmol·L-1 curcumin group were significantly increased(P<0.05 or P<0.01),and the expression level of Bcl-2 protein was significantly decreased(P<0.05);compared with shNC-C4-2+20 μmol·L-1 curcumin group,the expression levels of Bax and cleaved Caspase-3 proteins in the cells in shCD147-C4-2+20 μmol·L-1 curcumin group were significantly decreased(P<0.01);compared with shNC-LNCaP group,the expression levels of Bax,cleaved Caspase-3,and PARP1 proteins in the cells in shNC-LNCaP+20 μmol·L-1 curcumin group were significantly increased(P<0.05 or P<0.01),and the expression level of Bcl-2 protein was significantly decreased(P<0.05);compared with shNC-LNCaP+20 μmol·L-1 curcumin group,the expression levels of Bax,cleaved Caspase-3,and PARP1 proteins in the cells in shCD147-LNCaP+20 μmol·L-1 curcumin group were significantly decreased(P<0.05 or P<0.01),and the expression level of Bcl-2 protein was significantly increased(P<0.05).Compared with C4-2 group,the expression levels of E-cadherin protein in the cells in C4-2+20 μmol·L-1 curcumin group and C4-2+40 μ mol·L-1 curcumin group were significantly increased(P<0.01),and the expression levels of N-cadherin and Vimentin proteins were significantly decreased(P<0.01);compared with LNCaP group,the expression levels of E-cadherin protein in the cells in LNCaP+20 μmol·L-1 curcumin group and LNCaP+40 μmol·L-1 curcumin group were significantly increased(P<0.01),and the expression levels of N-cadherin and Vimentin proteins in the cells in LNCaP+40 μmol·L-1 curcumin group were significantly decreased(P<0.01);compared with shNC-C4-2 group,the expression levels of N-cadherin and Vimentin proteins in the cells in shNC-C4-2+20 μmol·L-1 curcumin group were significantly decreased(P<0.01);compared with shNC-C4-2+20 μmol·L-1 curcumin group,the expression level of E-cadherin protein in the cells in shCD147-C4-2+20 μmol·L-1 curcumin group was significantly decreased(P<0.01),and the expression levels of N-cadherin and Vimentin proteins were significantly increased(P<0.01);compared with shNC-LNCaP group,the expression level of E-cadherin protein in the cells in shNC-LNCaP+20 μmol·L-1 curcumin group was significantly increased(P<0.01),and the expression levels of N-cadherin and Vimentin proteins were significantly decreased(P<0.01);compared with shNC-LNCaP+20 μmol·L-1 curcumin group,the expression level of E-cadherin protein in the cells in shCD147-LNCaP+20 μmol·L-1 curcumin group was significantly decreased(P<0.01),and the expression level of N-cadherin was significantly increased(P<0.05).Conclusion:Curcumin inhibits the proliferation,migration,and invasion of the prostate cancer cells in vitro and induces the apoptosis;silencing the CD147 gene partially reduces its inhibitory effect and its ability to induce the apoptosis.

Objective ToassessthevalueofGdGEOBGDTPAenhancedT1ρimaginginevaluatingtheseverityandinflammation gradeinnonalcoholicsteatohepatitis(NASH)rabbitsmodel.Methods NASH modelswereestablishedin26adultrabbitsbyfeeding withthehighGfat,highGcholesteroldietinavarieddurations (0,4,8,12 weeks).T1ρ,T1ρinthehepatobiliaryphase (HBP)and changeofT1ρ(Δ%)werecomparedamongthedifferentgroupswhichweredeterminedbydifferentnonGalcoholicfattyliverdisease activityscore(NAS)andinflammationgrades.SpearmancorrelationanalysiswasusedtoassessthecorrelationsofT1ρ,T1ρ(HBP) withNASscoresandinflammationgrades.ROCcurvewasperformedtoevaluatethediagnosticvalueofT1ρ,T1ρ(HBP)inpredicting NASHandadvancedinflammation.Results T1ρandT1ρ(HBP)werepositivelyassociatedwithNASandinflammationscores.The differencesofT1ρ(HBP)amongNASH,nonalcoholicfattyliver(NAFL)andnormalliverwerestatisticallysignificant(P＜0.05). T1ρ(HBP)wassignificantlydifferentintherabbitswithgrade3inflammationfromintherabbitswithgrade0,grade1andgrade2 inflammation (P＜0.05).AUCsofT1ρandT1ρ(HBP)fordifferentiatingNASH were0.849and0.949,respectively.AUCofT1ρand T1ρGHBPfordiagnosinggrade2andgrade3inflammationwere0.925and0.922,respectively.Fibrosisandinflammationwerethe mainindependentfactorsaffectingT1(HBP).Conclusion GdGEOBGDTPAenhancedT1ρimagingcanreflecttheseverityofNASH anddegreeofinflammation.T1ρ(HBP)mightbeamoresuperiornoninvasiveimagingbiomarkerthannonGenhancedT1ρforassessmentof NASHactivityandinflammationgrading.

AIM:To investigate the effect of quercetin on the biological functions of rat bone marrow-derived endothelial progenitor cells (EPCs) and its potential mechanisms.METHODS:The bone marrow-derived mononuclear cells of Sprague-Dawley rats were isolated by density gradient centrifugation.The differentiated EPCs were cultured specially and stained with DiI-Ac-LDL and FITC-UEA-1.CD133+ and FLK-1+ were detected on the cell surfaces.After 14 d, the EPCs were incubated with a PI3K inhibitor BYL719 (3 μmol/L) and an ERK inhibitor FR180204 (15 μmol/L).After incubation of the inhibitors for 2 h, the cells were treated with quercetin at different concentrations (0, 10, 20, 40, 80 and 100 μmol/L).MTT assay and Transwell assay were used to detect cell viability and the number of migratory cells.The protein levels of AKT, eNOS, ERK and their phosphorylated status were determined by Western blot.RESULTS:Quercetin enhanced the viability and migration of the EPCs at a dose-dependent manner.However, the PI3K inhibitor BYL719 suppressed the QUE-induced cell viability and migration.Moreover, ERK inhibitor FR180204 exerted the similar inhibitory effect on the cell viability but had no effect on cell migration.Quercetin activated the phosphorylation of AKT, eNOS and ERK.On the other hand, BYL719 was observed to inhibit the phosphorylation of AKT and ERK.FR180204, however, was showed to inhibit the phosphorylation of ERK only.On the contrast, the stimulatory effects that quercetin exerted on the expression of eNOS and its phosphorylation were suppressed by BYL719 and FR180204.CONCLUSION:Quercetin stimulates the viability and migration of EPCs via PI3K/AKT/eNOS and ERK/eNOS signaling pathway, which would be beneficial for cardiovascular health.

AIM:To investigate the inhibitory effect of apolipoprotein A-I mimetic peptide D-4F on the scaven-ger receptor A1 ( SR-A1 ) in macrophage-derived foam cells induced by oxidized low-density lipoprotein ( ox-LDL ) . METHODS:RAW264.7 cells were pretreated with different concentrations (12.5, 25 and 50 mg/L) of D-4F or 50 mg/L inactive control peptide scrambled D-4F (sD-4F) for 1 h or endoplasmic reticulum stress (ERS) inhibitor 4-phenylbutyr-ic acid (5 mmol/L) for 30 min, followed by the treatment with 100 mg/L ox-LDL for 12 h.In addition, the cells were pre-treated with 50 mg/L D-4F or sD-4F for 1 h, and then stimulated with 2 mg/L tunicamycin (TM;an ERS inducer), for 4 h.The viability of the cells was measured by MTT assay, and the content of intracellular total cholesterol ( TC) was meas-ured by a tissue/cell TC assay.The protein and mRNA levels of SR-A1 and glucose-regulated protein 78 (GRP78) were analyzed by Western blotting and quantitative real-time PCR, respectively.The fluorescence intensity of DiI-ox-LDL in the cells was detected by a multifunctional microplate reader.RESULTS:D-4F significantly reduced ox-LDL-induced macro-phage injury and intracellular cholesterol accumulation, and attenuated the ox-LDL-induced expression of SRA1 and GRP78 in a dose-dependent manner.Additionally, D-4F significantly inhibited the TM-induced protein expression of SR-A1 and GRP78, and attenuated the uptake of ox-LDL by macrophages.CONCLUSION: D-4F reduces ox-LDL-induced macro-phage cholesterol accumulation and injury by inhibiting SR-A1 expression.The mechanism may be related to the inhibition of ERS signaling pathway mediated by GRP78.

Objective To observe the effect of Valpar 4 system on hand injury. Methods 40 patients with hand injury were randomly divided into control group and observation group with 20 patients in each group. Routine treatment was applied in both groups while Valpar 4 system was added in the observation group. Then total active motion (TAM) of finger joints, Disability of Arm Shoulder and Hand (DASH), and Upper Extremity Function Test (UEFT) were carried out before, 2 weeks and 4 weeks after treatment. Results The TAM improved significantly in the control group 2 weeks after treatment (P<0.001). The TAM, DASH and UEFT improved significantly in the observation group (P<0.001), and the DASH and UEFT were better in the observation group than in the control group (P<0.001). 4 weeks after treatment, the TAM and UEFT improved significantly (P<0.001) in the control group, the TAM, DASH, UEFT significantly improved in the observation group (P<0.001), and were better in the observation group than in the control group (P<0.05). Conclusion Valpar 4 system can effectively improve the TAM of fingers, function of hands and upper limb, especially in terms of activities of daily living of upper limbs.

Objective To explore the leaves water extract on the liver human hepatocellular carcinoma xenografts and protein expression of Bcl-2 and HSP70 .Methods From cell morphology and DNA agarose gel electrophoresis the observation leaves water extract the characteristics of the material role in human hepatoma cells ,further use of immunocytochemistry agent Western blotting analysis method oncogene Bcl-2 and tumor suppressor genes HSP70expression .Results After the processing of the the 60 mg/mL leaves water extract of liver cancer cells after 12 h ,liver cells showed severe shrinkage ,pyknosis ,highlight the cell plasma mem-brane ,immunohistochemistry and Western blotting analysis showed that during the experiment along with the oncogene Bcl-2 ex-pression weakened(P<0 .05) ,while the expression of tumor suppressor genes HSP70 had no obvious change(P<0 .05) .Conclusion Water extract of the leaves can be induced in vitro human liver cancer cell apoptosis ,and the apoptotic process may be followed by the oncogene Bcl-2 ,tumor suppressor gene expression of HSP70 relationship .

Objective To study the effect of early enteral nutrition (EEN) with ω-3 polyunsaturated fatty acid on nutritional status and body immunity of postoperative patients with colorectal cancer.Methods Forty patients with colorectal cancer were randomly divided into EEN group and parenteral nutrition (PN) group by systematic sampling method with 20 cases of each.The patients in EEN group were given complete enteral nutrition with ω-3 polyunsaturated fatty acid 1-7 d after operation,while the patients in PN group only were given nutrient by regular PN.Nutritional status,body immunity before operation and 1-7 d after operation,and intestinal function recovery time,untoward reaction were determined.Results The postoperative anus first exhaust time and defecate time of patients in EEN group were significantly shorter than those in PN group [(48.0±5.6) hvs.(51.0±6.7) h,(53.0±4.8) h vs.(72.0±3.6) h,P＜0.05].Serum albumin and prealbumin 1 d after operation in 2 groups were significantly lower than before operation (P ＜0.05),but 7 d after operation were significantly higher than 1 d after operation (P ＜0.05).Serum albumin and prealbumin 7 d after operation in EEN group were significantly higher than PN group(P ＜ 0.05).IgG,IgA,IgM,CD3,CD4,CD8,CD4/CD8 1 d after operation in 2 groups were significantly lower than before operation (P ＜ 0.05).IgG,IgA,CD4,CD4/CD8 7 d after operation in EEN group were significantly higher than PN group (P＜ 0.05).Conclusions Compared with the PN,EEN with ω-3 polyunsaturated fatty acid can remarkably improve nutritional status and body immunity in postoperative patients with colorectal cancer.It can accelerate the recovery of intestine function and worthy of recommending.