BACKGROUND:Several studies have demonstrated that the number of alveolar epithelial cells decreases gradually as pulmonary fibrosis develops,but the reasons remain unknown.The expression of P16,CyclinD1,and CDK4 might be abnormal and P16-CyclinD1/CDK4 call cycle regulatory pathway may play an important role in the progressive reduction of alveolar epithelial cells during pulmonary fibrosis.OBJECTIVE:To investigate the dynamic expression of P16,CyclinD1,and CDK4 in the pulmonary tissue of bleomycin-inducad pulmonary fibrosis mice.METHODS:Pulmonary fibrosis was induced in 6-week-old KM mice by intratracheal injection of bleomycin(BLM group).The control mice were administered the same volume of physiological saline(control roup).At 3,7,14,and 28 days after modeling,hematoxylin-eosin staining was performed to observe the pathomorphological changes of lung tissue.P16,CydinD1,and CDK4protein expression in lung tissue was detected by immunohistochemistry and Western blot analysis.RESULTS AND CONCLUSION:In the BLM group,typical changes of pulmonary fibrosis were observed,and P16 and CDK4protein expression in the pulmonary tissue increased with time,but CyclinD1 protein expression was decreased with time.P16and CDK4 protein-positive calls in the BLM group were more than in the control group.Compared with the control group,P16protein expression in the BLM group was higher at 14 and 28 days after modeling(P<0.01)and CDK4 protein expression was higher at 7,14 and 28 days after modeling(P<0.05).CydinD1 protein-positive cells and protein expression were greater at 3and 7 days after modeling in the BLM group(P<0,05),but they were less at 28 days after modeling(P<0.05),than in the control group.At 14 days after modeling,CydinD1 protein-positive cells in the BLM group were more,but CyclinD1 protein expression was ess,than in the control group(P<0.05).These findings suggest that P16,CyclinD1 and CDK4 protein expression was abnormal during pulmonary fibrosis and P16-CyclinD1/CDK4 cell cycle regulatory pathway greatly results in progressive reduction of alveolar epithelial calls during this process.

Objective To evaluate the application value of MiniFilerTM kit in LCN-STR genotyping in forensic science.Methods Compared the testing results of MiniFilerTM and IdentifilerTM kits in 49 general quantity biological samples and 39 LCN-biological samples,including blood stains,sperm stains,bones,and so on.Meanwhile the sensitivity of these two kits were also compared.Results Full concordance between IdentifilerTM and MiniFilerTM kits was observed in all of 49 general quantity biological samples.For 39 LCN-biological samples,only 22 samples could be genotyped in partial loci and 17 samples negative with the IdentifilerTM kit,while 30 samples could be genotyped in all loci,5 samples in partial loci and 4 samples negative with the MiniFilerTM kit.Therefore,there was a higher success rate for LCN-biological samples typing with MiniFilerTM kit than with IdentifilerTM kit.In addition,the sensitivity of the MiniFilerTMkit was also a little higer than the IdentifilerTMkit.Conclusion The results demonstrated that the MiniFilerTM kit can markedly improve the typing success rate of LCN-biological samples,and is suitable for analyzing difficult biological samples in forensic practice.

Objective To explore the role of insulin-like growth factor-2/insulin-like growth factor1 receptor/insulin receptor substrate-1 (IGF2/IGF1R/IRS1) signal pathway inducing the chemoresistance of epidermal growth factor receptor 2 (ErbB2) positive breast cancer cells to Herceptin.Methods Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot assay were used to determine the expression levels of IGF2,IGF1 R,and IRS1.The direct targets of miR-126 were validated by dual-luciferase reporter gene assay.In SKBR3/pool2 cells,IGF1 R activity was reduced by an inhibitor of IGF1 R,and IRS1 was knocked-down by shRNAs.Furthermore,3-(4,5-dimenthylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay was performed to evaluate the sensitivity of these treated cells to Herceptin.Results IGF2,IGF1 R,and IRS1 were significantly higher expressed in SKBR3/pool2 cell compared to that in SKBR3 cell.Western blot assay showed that IGF2/IGF1R/IRS1 was activated in SKBR3/pool2 cells.Bioinformatics analysis combined with luciferase activity suggested that miR-126 directly targeted IRS1.MTS results demonstrated that the chemosensitivity to Herceptin of SKBR3/ pool2 cells with inhibitor of IGF1R or shRNAs targeting IRS1 or overexpressing miR-126 was significantly reduced.Conclusions IGF2/IGF1R/IRS1 signal pathway confers to the chemoresistance of ErbB2 positive breast cancer cells to Herceptin.

[ ABSTRACT] AIM:To explore the expression pattern of microRNA-205 ( miR-205) in glioma tissues and its role in the invasion of glioma cells.METHODS:The expression of miR-205 and TBX18 was detected by real-time PCR and immunohistochemical observation, respectively.Transwell assay was used to examine the invasion change of U251 glioma cells after miR-205 overexpression via miR-205 mimics or decrease in miR-205 expression by miR-205 inhibitor.The target of miR-205 was searched by bioinformatics analysis combined with experimental analysis.The protein level of TBX18 was determined by Western blotting after siRNA transfection and Transwell assay was conducted.RESULTS:miR-205 expres-sion was downregulated in 82.6%of detected glioma tissues and TBX18 was significantly overexpressed in glioma tissues compared with normal tissues.miR-205 overexpression remarkably inhibited the invasion potential of U251 glioma cells with a decrease in the invasive cells (P<0.01), while inhibition of miR-205 significantly enhanced the invasion ability of U251 cells.Mechanically, miR-205 directly targeted TBX18 and downregulation of TBX18 also significantly inhibited the invasion potential of U251 cells with a decrease in the invasive cells ( P<0.01 ) .CONCLUSION: miR-205 expression is de-creased in glioma, and miR-205 inhibits glioma cell invasion via targeting TBX18.Our research contributes to the mecha-nisms responsible for glioma invasion and provides theoretical base for developing new therapeutic strategy to treat glioma.

Objective To evaluate the usefulness of diffusion-weighted MR combined with routine T2 WI in finding the possible residual foci in uterine cervical cancers after radical chemo-radiation therapy.Methods This was a retrospective study including 25 consecutive cervical cancer patients who received hysterectomy after radical chemo-radiation therapy.All of them underwent MR examinations post-chemoradiation and just before operation.Images of T2 WI alone and those of T2 WI combining DWI were evaluated respectively by 2 senior radiologists,in order to decide whether there were residual tumors.ADC values were also measured.Taking the post-operation pathological results as the gold standard,the accuracies,sensitivities and specificities of T2 WI alone,T2 WI combining DWI,and ADC values were all calculated.Results In those 25 patients,9 were found with foci of residual cancer in operative pathology,while no cancer cells were found in the other 1 6 patients.The accuracy,sensitivity and specificity in finding the positive residual cancer using T2 WI alone were 56.0%,77.8% and 43.8%,comparing with 72.0%,66.7% and 75.0% in T2 WI combining DWI.The accuracy and specificity increased with statistical significance after combining DWI (P =0.01 6 for accuracy,P =0.031 for specificity),while the sensitivity decreased but did not reach statistically significant level (P =0.099).No difference in ADC values was found.Conclusion DWI can be used as a supplementary sequence in finding the existence of residual tumors of cervical cancer after radical chemo-radiation therapy.Routine T2 WI combing DWI increased the specificity and accuracy,but still facing the risk of decreasing sensitivity.

] Objective To explore the role and mechanisms of FGF2 in chemo?resistance in breast cancer. Methods The inhibitors for different signal pathway were used to treat two drug?resistant breast cancer cell lines MCF?7/5?Fu and T47D/5?Fu established in our lab. MTS assay was used to determine chemo?sensitivity and Hoechst stain was used to measure apoptosis. Protein activation and FGF2 protein level in cell culture medium were detected by western blot and ELISA respectively. Results Akt inhibitor MK?2206 (20 nM) and mTOR inhibitor AZD8055 (2 nM) significantly reversed the chemo?resistance of MCF?7/5?Fu and T47D/5?Fu cell lines to 5?Fu and paclitaxel, but ERK1/2 inhibitor SCH772984 showed no significant effect. Compared to parent cell lines MCF?7 and T47D, p?Akt and p?S6K (represented as mTORactivity) levels were obviously up?regulated in MCF?7/5?Fu and T47D/5?Fu cell lines, and so do the FGF2 mRNA level and FGF2 protein level from culture medium. Moreover, FGFR inhibitor AZD4547 (4 nM) markedly reversed the chemo?resistance of MCF?7/5?Fu and T47D/5?Fu cell lines to 5?Fu and paclitaxel and down?regulated activation of FGFR?Akt?mTOR signal pathway. In agreement, FGF2 protein (10ng/ml) enhanced the chemo?resistance of MCF?7 and T47D cell lines to 5?Fu and paclitaxel and up?regulated activation of FGFR?Akt?mTOR signal pathway. Conclusion Activation of FGF2?FGFR?Akt?mTOR signal pathway promoted chemo?resistance of breast cancer cells.

Objective To investigate the impact of negative perfectionism on negative emotion(anxiety,depression) and the moderating effects of positive perfectionism.Methods A study was designed and a sample of 380 college students completed questionnaires including PANPS,SAS and SDS.Results ① Correlation analysis indicated positive perfectionism was negatively correlated with anxiety and depression (r1 =-0.25,P1 ＜0.01,r2 =-0.29,P2＜0.01),while negative perfectionism was positively correlated with anxiety and depression(r1 =0.26,P1 ＜0.01,r2 =0.22,P2＜0.01).② Moderating effects analysis indicated that positive perfectionism significantly moderated the regulating effect of negative perfectionism upon negative emotions(anxiety,depression) (β 1 =-2.64,β2 =-7.67,P＜0.01).Conclusion These findings suggest that the higher level of positive perfectionism,the greater influence of perfectionism on depression and anxiety,and positive perfectionism could buffer the negative perfectionism on anxiety and depression.

Objective To evaluate the effects of curcumin on the expression of phosphorylated extracellular signal-related kinase (p-ERK) and phosphorylated cAMP response element binding protein (p-CREB) in the spinal dorsal horn and dorsal root ganglion (DRG) in type 2 diabetic neuropathic pain (DNP) in rats.Methods Type 2 diabetes mellitus was induced by high-fat and high-sucrose diet and intraperitoneal streptozotocin (STZ) 35mg/kg,and confirmed by fasting blood glucose level≥ 16.7 mmol/L in male Sprague-Dawley rats.Type 2 DNP was confirmed by the mechanical withdrawal threshold (MWT) and thermal withdraw latency (TWL) measured on day 14 after STZ administration ＜ 80％ of the baseline value,and the rats with type 2 DNP were randomly divided into 3 groups (n =27 each):type 2 DNP group (group DNP),curcumin group (group Cur) and solvent control group (group SC).Curcumin and corn oil 100 mg/kg (25 mg/ml) were injected intraperitoneally once a day for 14consecutive days starting from 14 days after administration of streptozocin in Cur and SC groups,respectively.Another 27 normal rats were served as control group (group C) and were fed with common forage.MWT and TWL were measured at 3,7 and 14 days after curcumin injection (T1 3),and the lumbar segment 4-6 of the spinal cord and DRGs were removed at the same time for determination of the expression of p-ERK and p-CREB (by Western blot).Results Compared with group C,MWT was significantly decreased,TWL was shortened,and the expression of p-ERK and p-CREB in spinal dorsal horn and DRGs was up-regulated at T1-3 in DNP and SC groups,and at T1 in Cur group (P ＜ 0.05).Compared with group DNP,MWT was significantly increased,TWL was prolonged,and the expression of p-ERK and p-CREB in spinal dorsal horn and DRGs was down-regulated at T2,3 in Cur group (P ＜0.05).There was no significant difference in the MWT,TWL and expression of p-ERK and p-CREB between DNP and SC groups (P ＞ 0.05).Conclusion Curcumin can attenuate type 2 diabetic DNP by inhibiting up-regulation of the expression of p-ERK and p-CREB in the spinal dorsal horn and DRG in rats.

Objective To investigate the in vitro effects of quercitrin on the proliferation and the cytotoxicity of human γδT cells.Methods Peripheral blood mononuclear cells (PBMCs) were isolated from healthy subjects and cultured with isopentenyl pyrophosphate and IL -2 to induce human γδT cells.The hu-manγδT cells were cultured with quercitrin at various concentrations for 48 hours.CCK-8 kits were used to analyze the in vitro proliferation and cytotoxic activities of γδT cells.Flow cytometry was performed to meas-ure the expression of granzyme B and perforin in γδT cells.The expression of p-ERK, p-Akt and Bcl-2 at protein level were detected by Western blot .Results The percentage of human γδT cells in PBMCs was in-creased from (2.96±1.83)%to (88.94±2.36)%after 10 days of culture.The quercitrin at concentrations of 10 to 80 μg/ml could promote the growth of γδT cells and up-regulate the expression of granzyme B , per-forin, p-ERK, p-Akt and Bcl-2 in a dose dependent manner .The cytolytic activities of γδT cells against co-lonic carcinoma cells ( HCT116 ) were enhanced by quercitrin .Conclusion Quercitrin could promote the proliferation of γδT cells and enhance the expression of granzyme B and perforin at certain concentrations in vitro.ERK1/2 and Akt signal transduction systems might be involved in the process .

Objective:To investigate the mechanisms of TWS119 induced CCR5 expression in hunman γδT cells. Methods:After treatment with various concentrations of TWS119 for 48h, the expression of CCR5 in γδT cells were detected by flow cytometry. The p-STAT3 and GAPDH expression were examined by Western blot analysis. Results: TWS119 could upregulate the expression of CCR5 in dose dependent manner. Western blot analysis revealed that TWS119 inhibit phosphorylation of STAT3,but had no significant impact on GAPDH. In addition, pretreatment of γδT cells with 0. 5 μmol/L STAT3 specific phosphorylation inhibitor Stattic could upregulate the expression of CCR5 and enhance the TWS119 induced CCR5 expression. Conclusion: TWS119 could upregulate CCR5 expression of γδT cells by inhibiting STAT3 phosphorylation in vitro.