Objective:To study the influence of triptolide on neutrophils asthmatic mice of WBC and EOS in bronchoalveolar lavage fluid.Methods:Using ovalbumin ( OVA) combined with lipopolysaccharide ( LPS) method to establish sensitized asthmatic mice,BALB/c mice were randomly divided into 32 neutrophilic asthma group ( NA group ) , neutrophil triptolide intervention group (TLN group),neutrophil dexamethasone group (DXN group) and normal control group (NC group),n=8,hemocytometer calculated for each group of mice bronchoalveolar lavage fluid ( BALF) of the total number of WBC and EOS;smears stained Switzerland View in-flammatory cell infiltration.Results: In bronchioalveolar lavage fluid, numbers and infiltrations of WBC and EOS were significantly decreased in the DXN,TLN group than those in the NA group(P<0.05);but were significantly higher than the NC group (P<0.05), the DXN group above parameters were significantly higher than the TLN group ( all P<0.05 ) .Conclusion: Triptolide can reduce the total number of BALF WBC and EOS,inhibit lung WBC,EOS infiltration,ease neutrophilic airway inflammation.

Objective To investigate the effect of exogenous small RNA (dsP21-397) transfection on growth of human renal clear cell carcinoma cell lines A-498 and Caki-1. Methods The dsControl(control group) and dsP21-397(experimental group)were transfected into A-498 and Caki-1,respectively. The expression of p21 mRNA was analyzed by qRT-PCR. The expression of p21 protein,CDK4 and Cyclin D1 proteins were detected by Western blot. The cell cycle distribution was examined by flow cytometry(FCM). MTS assay and colony formation assay were used to analyze cell viability and proliferation ability. Results Compared with dsControl,p21 mRNA levels in A-498 and Caki-1 cells increased to 2.55-fold(P<0.01)and 2.18-fold(P<0.01),respectively,after transfection with dsP21-397. The expression of p21 protein was up-regulated while the expression of CDK4 and CyclinD1 were down-regulated. The percentage of cells in G0/G1 phase significantly increased after transfection of dsP21-397,and the proportion of cells in S phase and G2/M phase significantly decreased. The activity of A-498 and Caki-1 cells significantly decreased and the number of colonies in the dsP21-397 group was significantly lower. Conclusions dsP21-397 can significantly activate p21 protein expression,down-regulate the cell cycle-associated proteins,and inhibit the growth of renal clear cell carcinoma cells.

Objective To investigate the effect of microRNA-1180 transfection on the growth of renal cell carcinoma lines 786-O and ACHN.Methods The renal carcinoma cells were divided into the two groups:control group (transfecting dsControl) and experimental group (transfecting miR-1180).The expression change of p21 mRNA was detected by qRT-PCR.Western blot was conducted to analyze the expression changes of p21,CDK4,CDK6 and CyclinD1 proteins.Flow cytometry (FCM) was used to detect the cell cycle change.The MTS assay was conducted to detect the cell viability and the colony forming assay was performed to examine the cell proliferation ability.Results The qRT-PCR results showed that compared with the negative control dsControl group,after miR-1180 transfection,the expression level of·p21 mRNA in 786-O and ACHN cells was up-regulated to 2.54-fold and 2.49-fold respectively(P＜0.01).The expression trend of p21 protein was consistent with qRT-PCR results.The expression of CDK4,CDK6 and CyclinD1 proteins were significantly down-regulated.The FCM results showed that the proportion of cells in G0/ G1 phase was significantly increased after transfection of miR-1180,but the proportion of cells in S phase and G2/M phase was decreased significantly,indicating that the cell cycle was arrested in G0/G1 phase.The MTS assay results showed that compared with the dsControl group,the viability of the two kinds of renal carcinoma cells was significantly decreased.The colony formation assay showed that the number of colonies formed in the miR-1180 group was smaller,indicating the proliferation ability of miR-1180 transfected cells was decreased.Conclusion miR-1180 can significantly activate the p21 protein expression and inhibit the growth of renal carcinoma cell lines 786-O and ACHN.

Objective: To study the effects of microRNA-1180-5p (miR-1180-5p) on malignant biological behaviors of prostate cancer VCAP and LNCaP cells and the possible mechanisms. Methods: dsControl (dsControl group) and miR-1180-5p (miR-1180-5p group) were constructed and then transfected into two prostate cancer cell lines VCAPand LNCaP. qPCR and Western blotting were used to analyze the changes in mRNA and protein expressions of CDKN1A, Cyclin D1 and CDK6 after transfection. Cell cycle distribution, proliferation activity, clone formation capacity, cell migration and invasion ability were detected by flow cytometry, MTT assay, colony culture assay and Transwell assay, respectively. Results: qPCR results showed that compared with dsControl, CDKN1A mRNA levels in VCAP and LNCaP cells transfected with miR-1180-5p were up-regulated significantly, while the mRNA expressions of Cyclin D1 and CDK6 were significantly down-regulated (all P<0.01). Western blotting result was consistent with that of qPCR. The percentage of cells in G0/G1 phase was increased after transfection of miR-1180-5p (P<0.05), but the proportion of cells in S phase and G2/M phase was decreased and the cell cycle was arrested at G0/G1 phase (P<0.05). The proliferation activity of the two prostate cancer cells was significantly lower than that of the dsControl group after miR-1180-5p transfection (P<0.05), and the number of colonies in the miR-1180-5p group was significantly lower than that in the dsControl group (P<0.01). In the meanwhile, the cell migration and invasion ability in miR-1180-5p group was decreased (P<0.01). Conclusion: miR-1180-5p can significantly activate CDKN1A gene expression in prostate cancer cells and further inhibit the proliferation, migration and invasion of prostate cancer cells.