Objective: To detect the expression and distribution of activated cytotoxic cells in types of lymphoma with tissue microarray,and provide evidences for clinical treatment and prognosis. Methods: Immunohistochemical staining by S-P technique was used for detecting the expression and distribution of perforin and granzyme B in lymphoma tissue microarray,composed of 60 samples of lymphoma tissue.10 NK/T-cell lymphoma routine sections were used for relative research,and 10 reactive hyperplasia were used for comparison. Results: In the tissue microarray,samples originated from intranode and extranode were 48 and 12,respectively;consisting of 42 B-cell lymphoma,16 T-cell lymphoma(10 PTCLs,2 NK/T-cell lymphomas,2 lymphoblastic T-cell lymphomas,2 anaplastic large cell lymphomas),2 Hodgkin's disease.42 samples of B-cell lymphoma cells were negative in perforin and granzyme B.In 10 samples of peripheral T-cell lymphoma,perforin and granzyme B positive were 8 and 9,respectively,but the positive cells were no tumor cells.In 12 samples of NK/Tcell lymphoma(2 in the tissue microarray,10 routine sections),both perforin and granzyme B were strongly positive.B-cell lymphoma,T-cell lymphoma and NK/T-cell lymphoma differed significantly(P

Most of the clinical teachers in medical colleges have not received training in the teaching profession.This article explores the post-job education system of the clinical teachers in our hospital,aiming at improving the quality of medical education.

Diabetic retinopathy (DR) is one of the signs of complications related to diabetes in eye In clinic,there has not perfect measurement and method to prevent and treat DR.Proteomics as an emerging subject focuses on studying the organizational structure and related functions as a whole level.The application of proteomics research method can fully reveal the protein expression in tissues and cells with comparison of the difference between the normal and diabetes.The differentially expressed proteins can reveal the various factors in the pathological process of DR to address the accurate regulatory mechanism,and can find some new regulatory proteins associated with DR.This approach provides further theory and methodology to study DR,which has its unique advantages in drug-targeted treatment of DRs.

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 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 study the change and clinical value of SP‐D ,CCL18 and CC16 in serum and in exhaled breath con‐densate in acute exacerbation of chronic obstructive pulmonary disease .Methods Sixty two cases of COPD patients admitted in our hospital from 2010 January to 2013 December were selected as the research object .All the 62 patients were divided into group A(32 patients with COPD in acute exacerbation) and group B(30 patients with COPD in remission stage) in accordance with the severity of COPD .Thirty six cases of health people were selected as the control group .Statistical subjects SP‐D ,CCL18 ,CC16 content in se‐rum and in exhaled breath condensate ,and the relations between the various indexes and age ,smoking ,pulmonary function and BMI were analyzed .Results The exhaled breath condensate SP‐D ,CCL18 content in group A was significantly higher than that of B group and the control group (P<0 .05) ,and the SP‐D ,CCL18 in group B was higher than that in control group (P<0 .05) .The se‐rum SP‐D ,CCL18 ,CC16 content in group A was significantly higher than that of B group and the control group (P<0 .05) ,and the SP‐D ,CCL18 ,CC16 in group B was higher than that in control group (P< 0 .05) .Serum SP‐D ,CCL18 levels were significantly higher than those in the exhaled breath condensate (P< 0 .01) .Exhaled breath condensate SP‐D was positively associated with smoking age (r=0 .298 ,P<0 .05) ,and FEV1% pred ,FEV1/FVC (% ) showed a negative correlation (r= -0 .318 ,-0 .402 ,P<0 .05);the serum levels of SP‐D was positively associated with tobacco (r=0 .297 ,P<0 .05) ,and FEV1% pred ,FEV1/FVC (% ) were negative correlated (r= -0 .278 ,-0 .298 ,P<0 .05);serum CC16 and FEV1% pred ,FEV1/FVC (% ) were negatively corre‐lated (r= -0 .358 ,-0 .382 ,P<0 .05);Exhale breath condensate SP‐D ,condensate CCL18 ,SP‐D ,CCL18 serum ,serum CC16 were are positively correlated in each two (P<0 .05);and there was no significant correlation between other indexes and age ,smoking , pulmonary function and BMI etc .Conclusion Exhaled breath condensate ,serum SP‐D ,serum CCL18 ,exhaled breath condensate , exhaled breath condensate ,serum CC16 are closely related to acute exacerbation of COPD ,and monitoring the indicators can be judgment of the degree and prognosis of COPD .

Objective To construct an eukaryotic expression vector of pre-miR-15a,and to investigate the inhibitory effect of pre-miR-15a to Raji cells. Methods The pGCSIL-GFP vector encoding pre-miR-15a nucleotides was transfected into the bacterial competent cells,and then confirmed by PCR and sequencing analysis. The identified vector was transfected into Raji cells with oligofectamine 2000. The cells were divided into 3 groups,blank,negative control and pre-miR-15a group. Semi-quantitative RT-PCR was used to detect the expression of Bcl-2 mRNA,and immunofluorescence indirect for Bcl-2 protein expression. The growth of Raji cells was assayed by trypan blue dye exclusion method. Results PCR and sequences analysis indicated that the recombinant clones was identical with target sequences. Many green fluorescent cells were observed under fluorescent microscopy. The levels of Bcl-2 mRNA at every group had no obviously difference. Bcl-2 protein expression was obviously decreased at pre-miR-15a group compared with the other groups. Trypan blue dye exclusion method showed the cell growth was inhibited at 48 h and 72 h post-transfection. Conclusion We successfully construct the eukaryotic expression vector of pre-miR-15a,and it can inhibit the growth of Raji cells.

Objective To explore the nurses'self-efficacy for palliative care and its related factors.Methods Data were collected via a self-constructed questionnaire,using the purposive sampling method.Subjects were nurses from two “grade-A” general hospitals in Henan province.Results The nurses' self-efficacy for palliative care stayed at a disequilibrium state,mean score of self-efficacy for physical care was 4.00,followed by family care self-efficacy 3.85,then psychological and spiritual care self-efficacy 3.70.Significant difference existed in self-efficacy for palliative care in nurses having different attitudes toward death.The cognitive level for palliative care,past experience of caring for end-stage patients were positively correlated with self-efficacy.Conclusions The main factors of the nurses' self-efficacy for palliative care related to attitudes toward death,past experience of caring for end-stage patients and the cognition level of nurses to palliative care.

Aim To investigate effects of Cx26 on pro-liferation,apoptosis,migration of human highly meta-static hepatocellular carcinoma HCCLM3 cells.Meth-ods The HCCLM3 cells were infected with lentiviral vector targeting interference of Cx26 and the stable transfectants were selected by puromycin.The interfer-ence efficiency of Cx26 was detected by Real-time PCR and Western blot.Gap junction function was assessed by “parachute”dye-coupling assay. The effects of Cx26 interference on proliferation,apoptosis,migra-tion of HCCLM3 cells were determined by MTT assay, flow cytometry,transwell migration assay,respective-ly.Results The mRNA and protein expression of Cx26 in LV-Cx26 group was significantly lower than the LV-NC group and wide group (P<0.01),and GJ function in LV-Cx26 group also significantly reduced compared with LV-NC group and wild group (P <0.01 ).The Cx26 interference significantly inhibited the proliferation (P<0.01)),promoted the apoptosis (P <0.01 ),and decreased migration of HCCLM3 cells in vitro (P <0.01 ).Conclusion Lentiviral vector targeting interference Cx26 expression of HC-CLM3 cells can significantly reduce its GJ function,in-hibit the proliferation and migration,promote apopto-sis,and reduce its malignant properties.

Objective To optimize the extraction process of Ardisia crenata based on the content of total saponins and to provide a basis for the Ardisia crenata study. Methods Based on the results of the single-factor tests and the Box-Behnken central composite experimental design principles, a response surface methodology which has three factors and three levels was designed to optimize the extraction process of Ardisia crenata based on the content of saponins. Results A maximal extraction yield of total saponins reached 2. 29% under the optimal conditions as follows:70% alcohol was used as extraction solvent with the material to liquid ratio of 116, the extraction time was 3 h at 70 ℃. Conclusion The optimized extraction process is accurate, reliable and practically valuable.