Objective:To investigate the effects of 17β-Estradiol (E2) on mesenchymal stem cells (MSC) and to evaluate the effects of MSC treated with E2 on the maturation and function of dendritic cells (DC).Methods: We first isolated and cultured MSC from the human fetal lung.The MSC were treated with E2 for 24 hours at various concentrations ( 10~(-9),10~(-8) and 10~(-7) mol/L).After cell counting,proliferation,adherent ability and immunophenotypes of MSC were detected by flowcytometry.The gene expressions of cytokine (IL-6,TGF-βand VEGF) were measured by RT-PCR.The effects of MSC treated with E2 on the maturation and function of DC were determined.Results:After treated with E2,the proliferation and adherent ability of MSC were increased,while the immunophenotypes of MSC were not affected.When MSCs co-cultured with DC,MSC could inhibit the immuophenotypes and function of DC.However,when DC co-cultured with E2-pretreated MSC,the immunophenotypes (MHC Ⅱ,CD80 and CD86) of DC had been reconstructed.After treated with the high concentration of E2 for 24 hours,MSC secreted lower level of TGF-β than that in the control group,while IL-6 and VEGF expressions were increased compared with those in the control group.Conclusion: Estrogen may alter the immuno-suppressive effects of MSC on DC via modulating the cytokine secretion of MSC.

Objective To analyze the effect of GC-rich DNA fragments on the level of transgenic expression in Chinese hamster ovary (CHO) celts and its position effect.Methods The synthetic DNA fragment with GC-rich was cloned into the 5'or 3'or both 5'and 3'ends of expression cassette of expression vector.Three new expression vectors (pIRES-G1,pIRES-G2 and pIRES-G3) which was inserted with the GC-rich DNA fragments in different position were transfected CHO ceils,respectively,and then was observed under fluorescence microscope;the control vector was pIRES-EGFP.Stable transfected cell lines were screened under G418,and enhanced green fluorescent protein(EGFP) expression was analyzed by flow cytometry and the transgenic copy number was detected by quantitative real-time quantitative polymerase chain reaction (qRT-PCR).Results Three expression vectors with a GC-rich DNA fragments in different position were constructed successfully.The insertion of GC-rich DNA fragments at 3'end and both 5',3'ends of the box of expression vector could obviously improve the expression level of vector in CHO cells;and the expression level of the stably transfected CHO cells increased 1.39 fold and 1.32 fold compared to the control vector,respectively;the transgene copy number increased 1.32 fold and 1.24 fold compared with the control vector.While the insertion of GC-rich DNA fragments at 5'end of expression cassette had no obvious effect on the level of gene expression.Conclusion The role of DNA fragment with GC-rich in improving the transgenic expression of CHO cells is related to its position in the vector.The insertion of GC-rich DNA fragments at 3'end and both 5',3'ends of the box of expression vector can improve transgenic expression.

Objective To investigate the effect of different promoters on the expression level of transgene containing MAR expression vector in recombinant CHO cells.Methods The CMV promoter and 3-globin MAR were amplified by PCR,then CMV promoter was replaced the SV40 promoter in pCAT1 for constructing the expression vector droved by CMV promoter.The control vectors of pCAT1 and pCAT2 without containing MAR were simultaneously transfected into the CHO cells.Then the stably transfected cell line was screened by G418.The CAT gene expression level was analyzed by ELISA.Results The expression level of CAT enzyme in the cells transfected with MAR-containing vectors was increased compared with the cells transfected by pCATG and pCAT3 vectors without containing MAR,which were increased by 1.75 and 1.25 times respectively(P＜0.05);but CAT enzyme expression level in the pCAT1 transfected cells droved by SV40 promotor with the MAR-containing expression vectors was 1.4 times higher than that in the pCAT2 vector droved by the CMV promoter(P＜0.05).Conclusion MAR can enhance the transgene expression level in stably recombinant CHO cells,and the promoting efficiency of SV40 promoter and MAR combination is superior to that of CMV promoter and MAR combination.

By analyzing the questionnaire survey of consciousness of rights protection and status quo of rights and interests among medical staff in three 3rd degree and A grade hospitals located in northwest Hubei province,this paper concludes that medical staff there have a generally strong consciousness of rights,while their rights and interests are not well protected by society and hospitals,especially certain aspects including personal security,occupational health,working hours,legal vacation,and the rights to be involved in hospital management.It is suggested that legal rights and interests of medical staff should be improved through legislation,social support,and the direction function of public opinions.

OBJECTIVETo investigate the effects of glossy ganoderma spore oil on the proliferation, apoptosis, expression of miR-21 and its target genes of human lung adenocarcinoma SPC-A1 cell line, and to explore its possible mechanism.

METHODThe SPC-A1 cells were treated with glossy ganoderma spore oil for 24 and 48 hours. The inhibition growth efficacy was determined using cell count kit (CCK-8). Cell morphological changes were observed by light microscopy. Cell apoptosis was analyzed by flow cytometry. The expression of miR-21, PTEN and PDCD4 were determined by Real-time PCR.

RESULTGlossy ganoderma spore oil concentration-dependently inhibited the SPC-A1 cell's proliferation. When the concentration of glossy ganoderma spore oil attained to 0.2%, the cells' morphology changed obviously. Glossy ganoderma spore oil could induce the apoptosis of SPC-A1 cells at low concentration. Glossy ganoderma spore oil down-regulated the expression of miR-21 and up-regulated the expression of PTEN and PDCD4 significantly.

CONCLUSIONglossy ganoderma spore oil could inhibit the proliferation obviously and cause the changes of cell morphology. Furthermore, glossy ganoderma spore oil induced apoptosis of SPC-A1 cell through down-regulating the expression of miR-21 and up-regulating tumor suppressors.

Adenocarcinoma ; metabolism ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Ganoderma ; chemistry ; Humans ; Lung Neoplasms ; metabolism ; MicroRNAs ; genetics ; Polymerase Chain Reaction ; Spores, Fungal ; chemistry

Objective: To observe the effect of the respiratory rehabilitation and TCM exercises combined with conventional western medicine therapy for the patients with COPD. Methods: A total of 84 patients with COPD were randomized into the control group and observation group, 42 in each group. The control group were treated with conventional Western medicine treatment, and the observation group was treated with traditional Chinese exercises "sixth tactic" treatment on the basis of control group treatment. All patients were taken one year course of treatment. The changes in lung function, AECOPD occurrence, changes in symptom scores, Hamilton heart questionnaire integration were observed; and possible changes in the factors affecting the efficacy was analyzed by Logistic regression. Results: There were not significant differences in FEV1, FEV1/FVC, PEF and SpO₂ between the two groups before treatment. After 1 year of treatment, the FEV1 (1.58 ± 0.21 L vs.1.33 ± 0.22 L, t=8.092), FEV1/FVC (82.92% ± 8.42% vs. 62.81% ± 8.94%, t=10.013), PEF (358.27 ± 27.03 L/min vs. 324.13 ± 32.03, t=6.272), SpO₂ (99.58% ± 2.72% vs. 91.92% ± 2.89%, t=10.142) in the observation group were significantly higher than those in the control group (P<0.05). After treatment, Chinese medicine syndrome scores (6.3 ± 2.2 vs. 9.2 ± 2.0, t=4.652), HAMA score (6.0 ± 1.7 vs. 9.0 ± 2.0, t=4.563), HAMD score (6.1 ± 2.0 vs. 8.7 ± 1.7, t=5.094) in the observation group were superior to the subcontrol group (P<0.05). Regression analysis showed that there was a linear correlation between patient group, course of disease and outcome effect (P<0.05). Conclusions The respiratory rehabilitation and traditional Chinese medicine exercises "sixth tactic" combined with conventional treatment could release anxiety, depression and negative emotions, and improve clinical symptoms of the patients with COPD.

Objective To compare the effect of progressive teaching pattern with traditional long-term training on improvement of trainees ability of endoscopic ultrasonography ( EUS ) . Methods Ten learners who have achieved competence on routine endoscopic procedures but without any knowledge and skills on EUS procedures were enrolled in the study, and were randomly divided into two groups according to the sequence number. Learners with odd number received traditional long-term training, and the even number learners received progressive teaching. The proficiency of EUS knowledge and skills in the two groups were compared after one-year learning. Results At the final examination, the learners′ ability of mastering EUS was higher, and the study leave time was shorter in the progressive teaching group than that in the traditional long-term training group. however, we didn′t have a statistical comparison owing to the few numbers of trainees. Conclusion Progressive teaching pattern is superior to the traditional training, and it should be advocated when hospitals are relatively understaffed.

Objective To estimate the diagnostic value of cytology, DNA-ICM(DNA-image cytometry),cytology combined with DNA-ICM for pancreatic malignancy,and to explore the cut-off value for DNA-ICM. Methods Patients with suspicious pancreatic malignancy were retrospectively identified. In total,145 EUS-FNA specimens acquired from 140 separate patients were examined by cytology and DNA-ICM. Diagnostic values among cytology, DNA-ICM and the combination of the techniques in detecting pancreatic malignancy were compared. Results Compared with cytology, DNA-ICM had a lower sensitivity (63.0% VS 82.4%)and accuracy(69.7% VS 85.5%). After combining the techniques, the diagnostic value for pancreatic malignancy significantly improved compared with that by cytology(0.941 VS 0.912, P=0.007 0)or DNA-ICM only(0.941 VS 0.815, P<0.000 1). By using the Youden index, the cut-off value for DNA-ICM to detect pancreatic malignancy was one cell with DI(DNA index)≥2.5. Notably,with this standard, the sensitivity and accuracy of DNA-ICM significantly increased to 72.3% and 77.2%, and those of the combined techniques increased to 91.6% and 93.1%, respectively. Conclusion Automated DNA-ICM is an objective and effective method for pancreatic malignancy. Although DNA-ICM has a lower diagnostic value than that of conventional cytology, an improved value was obtained after combining the techniques.

Objective:To investigate the effect of S100 calcium-binding protein A9(S100A9)activation of nuclear factor kappa-B(NF-κB)on the upregulation of toll-like receptor 7(TLR7)expression and the release of inflammatory factors in microglia,as well as its underlying mechanism.Methods:The viability of BV2 microglia was assessed using CCK-8 kit.Transcriptome sequencing was employed to compare differential genes(DEGs)and identify target genes from the pool of differentially expressed genes.This analysis was complemented by GO analysis,KEGG enrichment analysis and the STRING database.The expression of TLR7 mRNA was verified by real time RT-PCR.The expressions of CD68 and CD206 were detected using immunofluorescence.The expressions of CD68,CD206,TLR7,p65,and p-p65 were detected using Western Blot.The level of interleukin 6(IL-6)and tumor necrosis factor alpha(TNF-α)were verified by ELISA.Results:Moderate concentrations of S100A9 had no inhibitory effect on microglial viability.Compared to the control group,the experimental group showed a significant increase in the expression level of CD68 pro-tein,while the CD206 protein was decreased.This suggests that S100A9 promotes the activation of BV2 microglia into pro-inflammatory types.TAK-242,an inhibitor of toll-like receptor 4(TLR4),significantly inhibited the expression levels of TNF-α and IL-6 after S100A9 stimulated BV2 cells.Activation of the TLR4/NF-κB pathway promoted the ex-pression of TLR7 protein.Conclusion:The moderate concentration of S100A9 can promote the polarization of microglia towards a proinflammatory direction.It also promotes the expression of TLR7 and the release of various inflammatory factors,including TNF-α and IL-6,through the activation of the TLR4/NF-κB pathway.This activation has an obvious proinflammatory effect.