Objective To investigate the inhibitory effect of siRNA targeting casein kinase 2 (CK2α)gene on the growth of HCT1 1 6 cells and to clarify its mechanism.Methods CK2α-siRNA sequence was designed according to mRNA sequence of CK2α. The in vitro cultured HCT1 1 6 cells were divided into normal control group (without transfection),negative control group (transfected with siRNA)and CK2α-siRNA group (transfected with CK2α-siRNA ),and the HCT116 cells were transfected with Lipofectamine 2000.The expression levels of CK2α,cyclin H,P53,and P21 proteins in the HCT116 cells were detected by Western blottting method,and the proliferation activities of the HCT116 cells were detected by MTT method,and the cell cycle was measured by flow cytometry. Results Compared with negative control group,the expression levels of CK2α,and cyclin H proteins in CK2α-siRNA group were decreased(P<0.01);the expression level of P53 protein had no change dramatically(P>0.05), and the expression level of P21 protein was increased significantly(P<0.01).Compared with negative control group,the survival rate in CK2α-siRNA group was decreased markedly 48 and 72 h after transfection detected by MTT method(P<0.01).Flow cytometry analysis showed the percent of the cells at G1 phase in CK2α-siRNA group was significantly higher than that in negative control group and the percent of the cells at S phase in CK2α-siRNA group was lower than that in negative control group(P<0.01),and the cell cycle was arrested at G1 phase. Conclusion siRNA targeting CK2αcan inhibit the proliferation of HCT116 cells and induce the arrest of G1 phase, which may be associated with inhibiting the expression of cyclin H and recovering the P53 activity after silencing CK2α.

Objective To study the expressions of casein kinase 2α(Ck2α),β-catenin, survivin in ovarian cancer tissue and analyze the relationships between them, and to explore the clinical application value. Methods Immunohistochemistry staining was used to detect the expressions of Ck2α,β-catenin and survivin in 8 cases of simple ovarian cysts tissue,8 cases of ovarian benign tumor tissue and 29 cases of ovarian cancer tissue;the correlations of the expressions of Ck2α,β-catenin,survivin in ovarian cancer tissue and their associations with the clinicopathologic characteristics were analyzed.Results The expression levels of Ck2α,β-catenin and survivin in ovarian cancer tissue were significantly higher than those in simple ovarian cysts tissue and ovarian benign tumor tissue(P<0.05);the Ck2αexpression level in ovarian cancer tissue was positively corelated with the expression levels ofβ-catenin and survivin(r=0.438,r=0.479,P<0.05);the expression levels of Ck2α,β-catenin and survivin in low-medium differentiation group were significantly higher than those in high differentiation group (P<0.05);the expression levels of Ck2α,β-catenin and survivin in stagesⅡandⅢ group were significantly higher than those in stage Ⅰ group(P<0.05).Conclusion The protein expression levels of Ck2α,β-catenin and survivin are increased in ovarian cancer tissue, and three are positive corelations between them;Ck2α,β-catenin and survivin may play an important role in the occurrence and development of ovarian cancer;the combined detection of them has important clinical value.

Objective To investigate the expression of protein kinase CK2α in thyroid carcinoma SW579 cells and its significance.Methods SW579 cells and Hacat cells were cultured in vitro and the expression levels of protein kinase CK2α mRNA and protein in SW579 cells and Hacat cells were measured by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting.Moreover,CK2 activity was also measured.Results It was showed that the mRNA (0.92 ±0.01 vs 0.52 ±0.02,t =19.24,P ＜0.01)and protein(0.98 ±0.01 vs 0.37 ±0.02,t =24.14,P ＜0.01) expression of protein kinase CK2αwere significantly stronger in SW579 cells relative to Hacat cells.Conclusions Formation and development of thyroid carcinoma may be in connection with the overexpression of protein kinase CK2α.

Objective We aimed to evaluatd the effect of oral nursing with oral washing on patients with peroral endotracheal intubation and discuss the appropriate oral nursing method for patients with patients with peroral endotracheal intubation. Methods We chose 150 patients with indwelling peroral endotracheal intubation after neurosurgery and divided them into the control group (73 cases) and the test group (77 cases). The test group received oral nursing with oral washing while the control group with routine oral nursing method. We compared the incidence of halitosis, the moist degree of oral cavity and the amount of the tooth fungus spot between the two groups. Results The incidence of halitosis and the amount of the tooth fungus spot in the test group were lower than those of the control group; The moist degree of oral cavity in the test group was higher than that of the control group, P<0.01. Conclusions Oral washing could effectively control halitosis, keep oral cavity moist and prevent the formation of tooth fungus spot.

Objective To detect the expressions of CD70 mRNA and protein and to determine the methylation status of CD70 gene promoter in T cells from patients with systemic lupus erythematosus (SLE).Methods Peripheral blood CD4+ and CD8+ T cells were isolated from 15 patients with active SLE,15 patients with inactive SLE and 15 healthy control subjects.Real-time quantitative reverse transcription-PCR was carried out to quantify the mRNA expression of CD70,flow cytometry to determine the frequency of CD4+CD70+ and CD8+ CD70+ T cells,and bisulfite sequencing to evaluate the methylation status of CD70 gene promoter in CD4+ and CD8+ T cells.Differences in these parameters among these groups were analyzed by one-factor analysis of variance and SNK-q test.Results Compared with the healthy controls,the patients with active SLE and inactive SLE showed a significant increase in CD70 mRNA expression in CD4+ T cells (0.82 ± 0.12 and 0.73 ± 0.11 vs.0.45 ±0.09,F =53.017,P ＜ 0.01) and in the frequency of CD70+CD4+ T cells (80.30％ ± 11.04％ and 66.80％ ± 3.98％ vs.12.48％ ± 3.45％,F =311.517,P ＜ 0.01).Also,the expression of CD70 mRNA in CD4+ T cells and the frequency of CD70+CD4+ T cells were significantly higher in patients with active SLE than in patients with inactive SLE (both P ＜ 0.05).There was a positive correlation between the frequency of peripheral CD70+CD4+ T cells and disease activity in SLE in these patients (r =0.792,P ＜ 0.01).The average methylation index of the region between-600 bp and-300 bp of CD70 gene promoter in CD4+ T cells was 0.32 ± 0.05 and 0.36 ± 0.05 respectively in the patients with active and inactive SLE,significantly lower than that in the healthy controls (0.62 ± 0.05,F =152.64,P ＜ 0.01),and the patients with active SLE showed a significantly lower methylation index than those with inactive SLE (P ＜ 0.05).Conclusions The CD70 gene promoter in CD4+ T cells is significantly hypomethylated in patients with SLE,which may directly lead to the overexpression of CD70.

Objective:To screen the new candidate molecules interacting with protein kinase Wee1B by yeast two hybrid system, and to analyze their interaction with Wee1B in the early stage of mouse fertilized eggs by bioinformatics.Methods:The plasmid pcDNA3.1/V5-His-TOPO-Wee1B wild type encoding mouse Wee1B gene was used as template to construct bait plasmid pGBKT7 Wee1B and the bait plasmid pGBKT7-Wee1B was transformed into yeast competent cells at SD/Trp (SDO),SD/Trp/X-α-Gal (SDO/X)and SD/Trp/X α Gal/AbA plates (SDO/X/A)plates to detect the toxicity and self-activation ability of yeast and its expression in yeast using Western blotting method.The yeast cells containing pGBKT7-Wee1B were fused with human ovary cDNA library, the yeast plasmid transformation of Escherichia coli positive clones were sequenced after identified by yeast transformation.BLAST analysis was carried out in GenBank,and its effect on the development of mouse fertilized eggs was deduced according to the gene annotation.Results:The double enzyme digestion analysis and sequencing analysis results showed that the pGBKT7-Wee1B bait plasmid was successfully constructed.The plasmid was transformed into the yeast,and there were no clones in the SDO/X/A plates.The pGBKT7-Wee1B and pGBKT7 empty vectors were transformed into the yeast,the bacteria were inoculated on the SDO plates,and the clones were uniformly grown on the two SDO plates.The positive clones were picked and expanded in culture,the protein was extracted and Western blotting showed that pGBKT7 Wee1B was expressed in the yeast.The bait plasmids were fused with human ovary cDNA library and the positive clones inserted into the fragment were identified by PCR. Nine proteins which interacted with Wee1B protein kinase were screened out by sequencing and blast analysis,and the proteins which could be closely related to the development of mouse oocytes and the development of fertilized eggs were analyzed by bioinformatics analysis.Conclusion:Using the yeast two hybrid system from human ovary cDNA library,nine interacting proteins with Wee1B protein kinase are screened and these screened proteins may regulate mouse oocyte maturation and early embryo development through interacting with Wee1B.

Objective:To understand the current situation of the demand for humanistic care of schizophrenics in convalescence and analyze the influencing factors, so as to provide the basis for clinical nurses to carry out targeted humanistic care for patients.Methods:A self-produced scale with a total score of 49-245 was used to investigate 516 convalescent period schizophrenic patients in 18 tertiary mental health institutions in 13 provinces of China, single factor analysis and multiple regression analysis were performed on their influencing factors.Results:The Likert 5-point scoring method was used. The overall score of the nursing care needs of hospitalized schizophrenic patients was (202.85±35.06) points, and the average score of total items were (4.14±0.16) points. Multiple linear regression results showed female ( B=0.100, P=0.024), stable marriage ( B=-0.098, P=0.026),high school (including secondary school) Education ( B=-0.107, P=0.018), family per capita monthly income of 2 000 yuan or above ( B=0.093, P=0.043),central region ( B=-0.110, P=0.014) were the main factors that affect the demand for humanistic care of inpatients with schizophrenia in convalescence. Conclusion:The hospitalized schizophrenic patients have higher and diversified needs for humanistic care. Patients with different genders, marital status, education level, monthly family income and living area have different needs for humanistic care. Individualized and standardized nursing should be given according to the actual situation of patients' needs to provide a more supportive environment for their rehabilitation.

Objective To investigate plasma NGF expression in adriamycin induced rat heart failure (HF) model .Methods Twenty‐five Wistar rats were randomly divided into the CHF group (n=15 ,adriamycin 4 mg/kg ,by intraperitoneal injection ,for 6 weeks) and normal control group (NC group ,n=10) ,after successful model construction ,the 6‐week observation was continuously conducted .The body mass and plasma NGF expression were detected once per 2 weeks .Results After 6 weeks later ,the body mass in the CHF group was significantly reduced ,the difference was statistically significant (P<0 .05) ,LVEDV and LVESV were sig‐nificantly increased ,while LVEF was declined obviously (P<0 .05) ,the NGF expression amount was significantly decreased com‐pared with the control group ,the differences were statistically significant (P<0 .05);the NGF expression amount was gradually re‐duced with the time extension of disease course(P<0 .05) .Conclusion Intraperitoneal injection of adriamycin can successfully in‐duce heart failure model in Wistar rats ,moreover NGF may be closely associated with HF .

Objective To investigate expression and significance of decorin(DCN)in liver tissue and serum of liver transplant patients with chronic rejection(CR).Methods Immunohistochemistry(SP method)was used to detect expression of DCN in liver tissue of 16 normal controls, 20 patients with cirrhosis, 46 liver translantion patients without CR and 8 patients with CR.Enzyme-linked immunosorbent assay method(ELISA method)was used to determined the content of DCN in serum of all research subjects.Results The expression of DCN was negative in normal hepatic tissues and with/without CR, cirrhosis tissues showed strong expression of DCN.The positive expression rate and the average optical density value of DCN in liver transplant tissues with CR had significant difference comparing with Cirrhosis tissues(25% vs 55%, 0.1249 ±0.0039 vs 0.2357 ±0.0396, P <0.01,while no statistic siqnificance compared to normal liver tissues and those without CR.The level of DCN in serum was significantly higher in liver transplant patients with CR, with significant difference comparing with normal people, liver cirrhosis and transplant liver patients without CR(54.0833 ± 6.0325)μg/L vs(1.0232 ± 0.9105)μg/L,(12.6202 ± 1.5370)μg/L,(17.7102 ± 2.3562)μg/L, P < 0.01).The concentration of DCN in serum showed a positive correlation with the degree of CR.Conclusions DCN showed negative expression in liver tissue and increased significantly in serum of liver transplantation patients with CR.This suggests that DCN may be involved in occurrence and development of CR.At the same time the determination of DCN in serum maybe become an important indicator of the early diagnosis, development and prognosis of CR for liver transplant patients.

Objective:To preliminarily explore the effects of tumor treating fields (TTF) arrays on the dose distribution in the treatment of Glioblastoma (GBM) using combined radiotherapy and concurrent TTF.Methods:EDR2 and MatriXX plate ionization chamber were employed to measure the absorbed doses of tissues at different depths (< 1 mm, 3 mm, 5 mm, 1 cm, 1.5 cm, 3 cm, 5 cm, 10 cm, and 15 cm) in the case that TTF arrays and latex-free foam were attached and not attached on the surface. Then the absorbed doses were calculated, compared, and analyzed. For the volumetric arc therapy (VMAT) of 10 GBM patients, deep dose verification was performed using the Sun Nuclear ArcCheck 3D dose verification system and the D99%, Dmean, and D1% of tumors and OARs were assessed. Results:The surface dose increased by 173% in the case that TTF arrays and latex-free foam were attached to the surface compared with the case of the surface with nothing attached. The surface dose increased by 61.7% due to the attachment of low-density latex-free foam. The dose deviation gradually decreased with an increase in the depth and stabilized (about 4%) at a depth of greater than 1.5 cm. As indicated by the VMAT verification result, the D99%, Dmean, and D1% of PTV and CTV decreased by 1.1%-1.2% and the Dmean and D1% of OARs (i.e., brainstem, pituitary gland, optic chiasma, optic nerve, eyeball, and eye crystal) decreased by 0.7%-1.5% in the case that TTF array and latex-free foam were attached on the surface compared with the case the surface with nothing attached. Conclusions:The combined radiotherapy and concurrent TTF in the GBM treatment will lead to a slight reduction of the absorbed doses of targets and OARs but a significant increase in the absorbed doses of the scalp. Therefore, it is recommended that the scalp doses should be reduced as far as possible in the design of the radiation treatment plan to reduce the adverse reactions on the scalp of GBM patients.