Objective To investigate the effect of knocking down miR-19a on proliferation and apoptosis of human glioma cell line U251 in vitro.Methods U251 cells were cultured routinely.MiR-19a inhibitor was transfected into U251 cells by Lipofectamine 2000.At the same time,the negative control group and blank control group were established.The expression level of miR-19a was detected by RT-PCR.and cell proliferation was analyzed by CCK-8 assay.The changes of cell apoptosis and cycle were monitored by flow cytometry.Results Compared with the negative control group and blank control group,qRT-PCR showed that the expression level of miR-19a was significantly reduced after transfection (F=124.72,P ＜ 0.01).CCK-8 revealed that proliferation ability of miR-19a inhibitor group was significantly suppressed.The cell survival rate in miR-19a inhibitor group after 48 h was (48.27-±8.23) ％,compared with the blank control group (100.00 ％),the difference was statistically significant (t =12.45,P ＜ 0.01).After low-expression of m iR-19a,the G0/G1 phase cells were increased and S phase cells were decreased.The low-expression of miR-19a could induce cell apoptosis.Conclusions Low-expression of miR-19a can inhibit cell proliferation,block G1/S transition and induce apoptosis in human glioma cell line U251.miR-19a may serve as an attractive target of gene therapy for glioblastoma.

ObjectiveTo investigate the level of fibrinogen in pleural effusion in tuberculous and malignant pleural effusion in content and nature of the diagnostic value of pleural effusion. MethodsCoagulation in STAGOcompact automated analyzer was determined to detect 42 cases of tuberculous pleural effusion and 38 cases of malignant pleural effusion levels of fibrinogen in patients with pleural effusion. ResultsThe level of fibrinopen in Tuberculous pleural effusion in malignant group was lover than that of tubeeculous group. Fibrinogen concentration of pleural effusion groups was significantly different( P ＜ 0.05). Thickenig in tuberculous group was greater than that in malignant pleural group(P ＜ 0.05). ConclusionThe determination of fibrinogen in pleural effusion had a considerable diagnostic value in differentiating tuberculosis from malignant pleural effusion.

Objective To investigate the diagnostic value and the clinical significance of six tumour markers of pleural effusion and serum in patients with lung cancer by detecting tumour marker and pleural effusion cytological examination. Methods Carcinoembryanic antigen (CEA), cancer antigen 19-9 (CA19-9),squamous cell carcinoma antigen (SCC), neuron specific enolase(NSE), cytokeratin 19 fragments(CYFRA21-l)and progastrin releasing peptide (ProCRP) levels in pleural effusion and serum sample from 50 cases of lung cancer and 30 cases of benign lung disease were detected by chemical and enzyme-linked immunosorbent assay.At the same time, the specimen of patients from pleural effusion was examinated by cytological method.According to the data above, the rational clinical diagnosis value were established by ROC curves. Results Six tumour markers in pleural effusion of lung cancer were higher than those of benign lung diseases, CEA,CA19-9, CYFRA21-1 and ProGRP were the highest group (P<0.05). The ratio of CEA, serum CYFRA21-1and CEA levels in pleural fluid and serum (P/S) in the group was the largest area under the ROC curve.Conclusion Pleural fluid CEA, the ratio of CYFRA21-1 and CEA levels in pleural fluid and serum (P/S) is helpful to differentiating benign and malignant pleural effusion diseases.The diagnostic value of pleural fluid CEA is important.

The human interleukin-24 gene (hIL-24),alse referred to as melanoma differentiation associated gene-7 (MDA-7), is a novel cancer-selective apoptosis-inducing eytokine gene with broadspectrum antitumor activity. Multiple data have demonstrated that treatment of tumors with a combination of IL-24 and other strategies such as drug therapy, radiation, dual gene therapy and so on, caused synergistic antitumor effects. These results will provide a impetus for further combination therapy in various tumors.

Objective To provide basis for identification and utilization of Phytolacca decandra. Methods Anatomic structures of different-aged storing roots, stems and leaves of P. decandra were studied by light microscope and histochemical localization. Results Different-aged vegetative organs have active proteins except the young roots, and the protein used to exist in parenchyma cells. ConclusionProtein content in young stems is the highest and the grain size is bigger than the other organs, and the protein content and grain size in young stems have notable differences from that in the old leaves, young leaves, old stems and roots. No protein has been found in two-year-old storing roots and extra cambium.

Objective:To investigate the effect of simvastatin on expression of pluripotent markers Oct3/4, Nanog, and Sox-2 in human breast cancer MCF-7 cells. Methods:Quantitative reverse transcription-polymerase chain reaction (qRT-PCR), immunofluo-rescent staining, flow cytometry, and Western blot were used to detect the expression of pluripotency markers Oct3/4, Nanog, and Sox-2 in human breast cancer MCF-7 cells treated with different doses of simvastatin. Results:qRT-PCR revealed the more signifi-cant inhibition of gene expressions of Oct3/4, Nanog, and Sox-2 in human breast cancer MCF-7 cells when subjected to high doses of simvastatin (10, 50, and 100 μmol/L) compared with the control group (P<0.05). By contrast, no significant difference was observed be-tween the expressions of low-dose simvastatin treatment (1 μmol/L) and control (P>0.05). The inhibitory effect of simvastatin on the gene expressions of Oct3/4 and Nanog was more significantly apparent at 50 and 100 μmol/L dosages than at 10 μmol/L (P<0.05). By contrast, no significant difference in the inhibitory expression of Sox-2 was observed among the three high-dose treatments (P>0.05). Between the two higher-dose treatments (50 and 100 μmol/L), no significant difference in the inhibitory expressions of Oct3/4, Nanog, and Sox-2 in MCF-7 cells was found. Meanwhile, in the 10 μmol/L simvastatin treatment, immunoflurescent staining showed a marked reduction in the protein expression of all three pluripotent markers in MCF-7 cells, and flow cytometry demonstrated a decrease of Oct3/4-, Nanog-, and Sox-2-positive cells (P<0.05). Western blot further revealed that the protein expression of Oct3/4, Nanog, and Sox-2 in MCF-7 cells was significantly declined by the same simvastatin dose (P<0.05). Conclusion: Simvastatin can inhibit the expression of pluripotent markers Oct3/4, Nanog, and Sox-2 in human breast cancer MCF-7 cells, proving the anti-cancer properties of simvastatin.

Objective:CⅡTA is a transcriptional factor for transactivation of HLA expression.The aim of the study is to investigate the regulation of prednisone on inducible expression of CⅡTA in native PBMCs.Methods:Normal PBMC was stimulated with combination of IFN-? and PHA with or without the presense of prednisone.CⅡTA mRNA in the cells was amplified by RT-PCR,for analyzing transcriptive level in three designed groups of native PBMCs,activated PBMCs using the biomodulators and the triggered PBMCs in presense of prednisoloen.Results:Normal human PBMCs constitutively expressed CⅡTA,which was increased by stimulation of PHA and IFN-?.Prednisone significantly inhibited the inducible pattern of CⅡTA expression.Conclusion:Prednisone has showed inhibition effect on the expression of PBMC CⅡTA by descending transcription of the specialized transcriptional factor in activated PBMCs,whereby to downregulate the expression of HLA in PBMCs.

OBJECTIVE To explore the gene gyrA mutation in nalidixic acid-resistant Salmonella paratyphi A(SPA) in Shaoxing area.METHODS Fifty-two strains of SPA gene were assessed by amplification and purification and the PCR products were detected by two directional sequencing methods.The sequences of DNA gyrase A were compared and analyzed with the OMIGA2.0 software,and the gene gyrA mutation was identified.RESULTS The mutation in nalidixic acid-resistant SPA was discovered in No.83 or No.87 animo acid site of gyrA quinolones resistant determinate region(QRDR),and the mutation of 51 strains occurred in No.83 site,1 strain occurred in No.87 site.The simultaneous mutation of two sites was not discovered,No mutations were found in other sites of gyrA QRDR.CONCLUSIONS No.83 site mutations of gene gyrA has a direct relation to nalidixic acid-resistant SPA.No.87 site mutation of gene gyrA might also cause nalidixic acid-resistantce.The nalidixic acid-resistant SPA is mainly mutated in No.83 site by TCC→TTC(Ser→Phe) in Shaoxing area.

Objective To investigate the synergistic proliferation-inhibiting and apoptosis-inducing effects of recombinant human-derived interleukin-24 (rhIL-24) and recombinant human-derived decorin (rhDCN) on human hepatocellular carcinoma cells HepG2.Methods Cellular growth and morphological changes of HepG2 cells were observed under the inverted microscope at 48 h after being transiently transfected with pcDNA3.1 (+)-IL-24 and pcDNA3.1 (+)-DCN by Lipofectamine.The proliferation-inhibiting effects of IL-24 and DCN on HepG2 cells,respectively and jointly,were observed with MTT assay at 24 h,48 h and 72 h post-transfection.Apoptosis and cell-cycle of HepG2 cells were analyzed by flow cytometry at 48 h post-transfection.Results Compared to control groups,the cells of target gene groups presented typically changes of proliferation inhibition and apoptotic morphology,which occurred obviously in co-transfection group.The results of MTT assay showed that at 48 h and 72 h post-transfection,the profiferation-inhibiting rates in the group of cells co-transfected with IL-24 and DCN were (31.88±6.57) ％ and (36.83±3.76) ％,respectively,displaying significant difference with those of other groups (P ＜ 0.01).The results of flow cytometry showed that IL-24 and DCN can induce HepG2 cells apoptosis to some extent.Compared to the early apoptosis rate of cells of control groups,plasmid (2.98±0.72) ％,blank cell (3.50±0.92) ％,IL-24 (20.01±1.08) ％ and DCN (22.20±0.91) ％,a statistically remarkable apoptosis rate,(32.56±0.90) ％,can be seen in the group of cells treated with IL-24 and DCN jointly (P ＜ 0.01).The result of cell cycle analysis revealed that,compared to control groups,the proportion of cells was higher in the phase of G2/M in the IL-24 group (11.24±0.35) ％ and in the phase of G0/G1 in the DCN group (77.93±0.67) ％.The proportions of cells in the phases of G2/M increased to (71.36±0.60) ％ and that of G0/G1 statistically increased to (10.39±0.67) ％ in the group of cells co-transfected with IL-24 and DCN (P ＜ 0.01).Conclusions Combinatorial treatment of HepG2 cells with IL-24 and DCN can exert stronger synergistic proliferation-inhibiting effect and apoptosisinducing activity-in comparison to single therapies.IL-24 and DCN can induce cell cycle arrest on HepG2 cells,occurred in the phase of G2/M and G0/G1,respectively.Promoting effect of cell cycle arrest in the phase of both G2/M and G0/G1 can be seen on HepG2 cells co-transfected with IL-24 and DCN,which maybe the possible mechanism of the synergistic proliferation-inhibiting and apoptosis-inducing effect.

Objective]Observe clinical efficacy of herpes zoster treated by the compound radix scutellariae solution wet application. [Method]50 patients were randomly divided into 25 cases of the compound radix scutellariae group and 25 cases of the nitrofurazone group, respectively given wet application of the compound radix scute-llariae solution and nitrofurazone solution, at the same time given intravenousganciclovir 0.3g/time,1 time/d, oral vitamin B1 20mg/time, 3 times/d, oral cobalt amine 0.5mg/time, 3 times/d, one week as a period. Record all patients' pain, skin lesion scores before the treatment and in the course of the third, fifth, seventh day and record the time of herpes stopping hair, blister drying up, blister beginning to scab and the scabby area not less than 50%.[Result] Pain, skin lesion scores of the compound radix scutellariae group were signifycantly lower than those of nitrofurazone group, the time of herpes stopping hair, blister drying up, blister beginning to scab and the scabby area not less than 50% of the compound radix scutellariae group compared with nitrofurazone group in advance, the difference was statistically significant(P<0.05). [Conclusion]Treatment of the compound radix scutellariae solution to herpes zoster had distinct clinical efficacy, having significant advantages in terms of alleviating pain and promoting skin lesions subsided.