Objective This study aims to investigate the post-transcriptional regulatory effects that control proliferation, apoptosis, and invasion in pediatriccholesteatoma keratinocytes.In particular, the potential role of miR-21was focused on in this study.Methods A total of 23 pediatric cholesteatoma tissues were processed for cell culture.Pediatriccholesteatoma keratinocytes were transfected with miR-21 inhibitors, or negative control miRNAs.RT-PCR was used to assess the expression levels of miR-21.EdU incorporation assay and TUNEL staining were used to assess the proliferation and apoptosis of pediatric cholesteatomakeratinocytes, respectively.Results MiRNA-21 was downregulated when pediatric cholesteatoma keratinocytes were transfected with miR-21 inhibitors.Furthermore, the number of proliferative EdU+cells decreased in cholesteatoma keratinocytes transfected with miR-21 inhibitors;and the number of TUNEL-positive cells also increased in cells transfected with miR-21 inhibitors, compared with cells transfected control miRNA.Conclusion MiRNA-21 promotes the proliferation and inhibits apoptosis of pediatric cholesteatoma keratinocytes.

Objective To explore the expressions of annexin and epidermal growth factor receptor(EGFR) in pediatric middle ear cholesteatoma.Methods Twenty-three children with middle ear cholesteatoma and 26 children with normal skin of external auditory canal(control group) were selected from the children enrolled in the First Affiliated Hospital of Zhengzhou University from August 2014 to March 2016.The expressions of annexin A1 (AnxA1),AnxA2 and EGFR mRNA in cholesteatoma and normal tissues were analyzed by quantitative real-time PCR (qPCR).Protein expressions of AnxA1,AnxA2 and EGFR were evaluated by using Western blot and immunohistochemistry methods.Results The expressions of AnxA1,AnxA2 and EGFR mRNA were significantly increased in cholesteatoma compared with the control group (AnxA1:4.68 ± 1.77 vs.2.65 ± 0.96,U =111.5,P ＜ 0.001;AnxA2:3.89 ± 1.00 vs.2.4 7 ± 0.81,U =84.5,P ＜ 0.001;EG FR:4.97 ± 1.85 vs.3.50 ± 0.95,U =15 3.5,P =0.004).AnxA1 and AnxA2 mRNA expressions were positively correlated with EGFR mRNA (AnxA1 and EGFR:r2 =0.283 2,P =0.009;AnxA2 and EGFR:r2 =0.213 5,P =0.027).Compared with the control group,protein expressions of AnxA1,AnxA2 and EGFR were markedly enhanced (AnxA1:0.450 ±0.031 vs.0.320 ±0.026,U =102.4,P ＜0.001;AnxA2:0.568 ±0.024 vs.0.365 ±0.028,U =94.6,P ＜0.001;EGFR:0.397 ±0.021 vs.0.228 ±0.017,U =128.4,P ＜0.001).The results of immunohistochemistry analysis showed that the ratio of AnxA1,AnxA2 and EGFR positive cells were higher than those in the control group(AnxA1:65.22％ vs.38.46％,x2 =9.296,P =0.026;AnxA2:69.57％ vs.46.15％,x2 =8.378,P =0.039;EGFR:69.57％ vs.50.00％,x2 =10.574,P =0.014).Conclusions The expressions of AnxAl,AnxA2 and EGFR are upregulated in pediatric cholesteatoma,with AnxA1 and AnxA2 expressions positively correlated with EGFR.

Objective To optimize multiplicity of infection ( MOI) and antibiotics ( blasticidin) concentration selecting BSD gene in construction of monoclonal stable cell line by lentivirus vector-mediated RNA interence silenced gene SGMS2.Methods The INS-1 cells were transfected by fluorescence labeled negative control SGMS2-siRNA lentivirus at MOI of 0, 10, 30, 60 and 120 TU number/cell.The cells were photographed under fluorescent microscopy after 72 h cultivation, then fluorescence ratio and apoptosis rate were calculated to determine optimal MOI.The INS-1 cells were treated by blasticidin with different concentrations of 0, 1, 2, and 3 μg/mL, and the apoptosis rate was observed to acquire optimal concentration of antibiotics.The INS-1 cells were transfected by negative control SGMS2-siRNA lentivirus and SGMS2-siRNA lentivirus (virus titer:1 ×108TU/mL) at optimal MOI and positive-transfected cells were selected by blasticidin at optimal concentration, then mixed cell lines were acquired.The monoclonal cell line was constructed at fluorescence ratio of 90%.Results The optimal MOI was 60 with 100% fluorescence ratio, less than 0.5% apoptosis rate and keep original cellular morphology.The optimal concentration of blasticidin was 2 μg/mL with cell adherence disappear and all cells apoptosis.The Ct value of INS-1-SEMS2 cells detected at the second time was 28.21, which was greater than 27.58 at the first time.The interfering efficiency of siRNA was 77.78% which indicated a successful expression of siRNA and construction of monoclonal stable cell line ( INS-1-SEMS2 ).Conclusion The monoclonal stable cell line was successfully constructed by lentivirus vector-mediated RNA interence silenced gene SGMS2.

Objective To discuss the effect of family nursing intervention on the life quality and pulmary function of patients with chronic obstructive pulmonary disease (COPD). Methods We divided 72 patients with COPD into the test group and the control group with 36 cases in each group.The two groups received routine treatment and nursing but additional family intervention was given to the patients and fam-ily members in the test group.The life quality and pulmonary function after intervention in the two groups were appraised in the two groups. Results The evaluation of life quality and pulmonary function were alleviated after intervention compared with those before intervention (P ＜ 0.05).But the above items in the control group were not significantly improved (P ＞ 0.05). Conclusions Effective family intervention could improve the life quality and pulmonary function of patients with COPD.

The current problems of pharmacology teaching in nursing in 3-year program colleges lie in that the teaching ideas drop behind;course orientation is vague;the development of text books lags;the contents lack characteristics;and there is no clear aims in the practice teaching.So,it is essential to make the teaching evolution.First,according to the post needs,the orientation of the courses should be definite,and new ideas about the curriculum should be set up.Meanwhile,the development of text books should be speeded up and the teaching contents should be optimized.Then,practice teaching needs to be improved,and cultivation of their comprehensive competence should be enhanced.To improve the students’professional qualities,emphasis on the penetration of humanities should be paid attention to.

OBJECTIVE: To observe the effects of glycyrrhizin on hydroxyproline(HYP),hyaluronic acid(HA) and laminin(LN) in pulmonary fibrosis model rats.METHODS: A total of 80 SD rats were randomly divided into normal control group,model group,hormone group(prednisone 0.6 mg?kg?d-1),and glycyrrhizin group(10 g?kg?d-1).The latter 3 groups were established into pulmonary fibrosis model with bleomycin,and on the second day after modelling,each group was given corresponding test drugs(the normal group and model group were treated with same amount of normal saline) then the changes of the levels of HYP in lung tissues,and serum levels of HA and LN in every group were observed at 7 and 28 days respectively.RESULTS: In glycyrrhizin group compared with model group,the contents of HYP in lung tissue,HA and LN in serum decreased significantly(P

OBJECTIVE: To explore the effect of glycyrrhizin on eotaxin expression in lung tissue from asthmatic model mice and the mechanism for glycyrrhizin to treat bronchial asthma.METHODS: The mice were randomized to asthma model group,prednisone-treated group and glycyrrhizin-treated group and normal control group.Egg protein asthma mouse model was established before being giving the corresponding drugs.Then the mice were sacrificed after 4 weeks for the counting of eosinophils in bronchoalveolar lavage fluid(BALF) and detection of Eotaxin expression in lung tissues by immunohistochemistry,with the results compared with normal control group.RESULTS: In glycyrrhizin-treated group compared with model group,the counting of eosinophils in BALF were decreased(P

OBJECTIVE To investigate the effect of nuclear factor-kappa B ligand-receptor activator of nuclear factor-kappa B-osteoprotegerin(RANKRANKL-OPG)system on bone resorption in middle ear cholesteatoma,and the reasons why middle ear cholesteatoma in children were more aggressive. METHODS The expression of RANKL and OPG in 21 adults cholesteatoma,17 children cholesteatoma and 18 normal external meatal skin was studied with immunohistochemistry. Semiquantitative analysis was used for comparison and statistical analysis. RESULTS RANKL-positive cells and the ratio of RANKL/OPG in cholesteatoma were significantly higher than that in external meatal skin(P

Objective:To analyze the association of type 2 diabetes mellitus(DM) with the clinical and coronary angiographic features of coronary heart disease(CHD) in patients aged over 70 years old.Methods: A total of 310 elderly patients with coronary angiograph-confirmed coronary diseases,who were treated in Changhai Hospital during Apr.2006 to Jul.2008,were retrospectively analyzed.The patients were further divided into 2 subgroups according to the presence of DM: DM-CHD group(n=155) and non-DM-CHD group(n=155).The age,gender,blood pressure,blood lipid,ejection fraction(EF),the angiographic outcomes,etc.were analyzed and compared between the two groups.Results: The incidence of hypertension was significantly higher in the DM-CHD group than in the non-DM-CHD group(P

60%(lot number 20040828, Jiahui, Shanghai, China).METHODS:Rabbit autologous BMSCs were isolated and cultured in vitro.At the third passage, BMSCs at 2?1010/L were combined on 3 cm?2 cm gelatin sponge.Each gelatin sponge contained 60 ?L cell suspension, placed in a cell incubator for 4 hours for further use.In each group, rabbit models of steroid-associated femoral head necrosis were established by liquid nitrogen frozen method.After natural rewarming, a hole was drilled.Rabbits in the model control group were left intact.Rabbits in the decompression group were implanted with gelatin sponge.Rabbits of the BMSC transplantation and theaflavin + BMSC transplantation groups were implanted with gelatin sponge with rabbit autologous BMSCs, and perfused with 250 mg theaflavin three time every day till death.MAIN OUTCOME MEASURES:The following parameters were measured:general conditions of animals;signal changes in the femoral head necrosis region were observed in the hip by magnetic resonance imaging scanning.Histology of femoral head samples was observed.Results of scanning electron microscope observation.RESULTS:After 8 weeks, rabbits became more active than before treatment, and walking way became normal gradually in the BMSC transplantation, theaflavin + BMSC transplantation groups.Four weeks after model induction, magnetic resonance imaging signal region of all groups had no obvious changes, but 8 weeks later, the necrosis signal region of the model control group magnified while it reduced obviously in the decompression group.Low signal region range became slightly small in the cell transplantation group.The low signal region range became significantly small in the theaflavin + BMSC transplantation group, and bone density signal was nearly normal.Compared with other three groups, positive number of empty lacunae significantly reduced in the theaflavin + BMSC transplantation group at 4 and 8 weeks following model induction(P