Purpose To construct a lentiviral ventor-mediated RNA interference of human midkine ( MK) gene and to provide the ba-sis for further experiment in vivo and in vitro. Methods Four RNAi sequences targeting human MK gene were designed, and cloned into the lentiviral vector to construct lentiviral vectors:GV115-MK-1, GV115-MK-2, GV115-MK-3 and GV115-MK-4. After transfec-tion into competent E. Coli bacteria, the candidate clones were identified by PCR and DNA sequencing. The titer of lentiviruses was determined after 293 T cells were co-transfected with GV115-MK, pHelper 1. 0 and pHelper 2. 0. The four kinds of recombinant lenti-viruses were used to infect human breast cancer cell MDA-MB-231, and the expression levels of MK mRNA were detected by real-time PCR. Results PCR analysis and DNA sequencing confirmed that the four inserted MK RNAi sequences were corrected. Strong green fluorescence was observed in the 293 T cells under the fluorescent microscope after co-transfection. The titer of the concentrated virus was 6 × 108 , 5 × 108 , 5 × 108 and 6 × 108 TU/ml, respectively. The MK expression in MDA-MB-231 cells infected with lentiviral vec-tors GV115-MK-1 was decreased by 87. 2% (P<0. 05). Conclusion Lentiviral RNAi vector for MK gene is successfully construc-ted, and it could effectively inhibit the expression of MK gene in MDA-MB-231 cells in vitro.

ObjectiveTo investigate the intervention of level of atorvastatin calcium tablets (lipitor) on soluble CD40 (CD40L) and high sensitivity C-reactive protein (hs-CRP) in patients with angina.Methods 65 patients with coronary heart disease were divided into stable angina (SA group,n= 30) and unstable angina (UA group,n= 35).The vein blood was collected in the hospitalized morning,2,4,6 weeks after treatment and serum levels of hs-CRP and CD40L were measured and compared between the two groups.ResultsThe levels of soluable CD40L [(20.52± 2.91)μg/L] and CRP [(7.96±1.69) mg/L] in UA group were higher than SA group [(7.96±-1.35) tg/L and ( 1.58 ± 0.91 ) mg/L] (t = 21.705、18.493,both P＜ 0.05) before treatment,and after treatment,their levels in the two groups were significantly lower than pre-treatment (P＜0.01).ConclusionsCD40L and hs-CRP may involved in the pathophysiology of unstable angina process and can be used as an indicator reflecting vulnerable plaque.Lipitor might enhance stability of atherosclerotic plaque and prevent acute coronary events by reducing levels of CD40L and hs-CRP.

Since the early symptoms are very hidden,most of patients with colorectal cancer (CRC)have occurred metastasis when diagnosed.Recently,chemotherapy combined with targeted therapy is the major therapeutic strategy for metastatic CRC and the high frequency activation of phosphatidylinositol 3-kinase (PI3K) signaling pathway has been validated sharing a close relationship with the development of CRC,contributing important values for the diagnosis and therapy of CRC.

Objective To investigate the adjustment and value of lymphocyte G protein-coupled receptor kinase-2 (GRK2) protein levels in patients with acute coronary syndrome(ACS). Methods Forty-two patients with stable angina pectoris (SAP), 44 patients with unstable angina pectoris (UAP), 43 patients with acute myocardial infarction (AMI), and 46 patients with normal coronary angiography (NCA) in hospital were enrolled in this study. Lymphocyte GRK2 protein levels were analyzed by Western blot in 24 h after admitted to hospital. Heart rate variability (HRV) analysis was performed based on 24 h Holter electrocardiogram (ECG) monitoring. Cardiac functions were measured using ultrasonic cardiogram. The results were compared and the relationship between GRK2 protein levels and HRV, cardiac functions index was analyzed. Results The level of lymphocyte GRK2 in AMI group, UAP group, SAP group and NCA group was (209.8 ± 63.9)%, (165.6 ± 60.2)%, (131.7 ± 51.8)% and (125.3 ± 50.6)%. The levels of lymphocyte GRK2 in AMI group and UAP group was significantly higher than that in SAP group and NCA group .Moreover, the level of GRK2 was the highest in AMI group, and there were significant differences (P<0.01). The level of lymphocyte GRK2 had negative correlation with high frequency(HF), low frequency(LF), LF/HF, standard deviation NN interval (SDNN), standard deviation of the average normal RR interval for 5-minute segments (SDNNI) and left ventricular ejection fraction (LVEF) (r =-0.52,-0.47,-0.53,-0.56,-0.49,-0.51, P < 0.01). Conclusions The rise of lymphocyte GRK2 protein levels is significantly associated with increased sympathetic nerve excitability and deterioration of cardiac function.

Purpose To observe the effect of MK on proliferation, migration and angiogenesis of human umbilical vein endothelial HU-VECs cells and to explore the role of MK in tumor angiogenesis. Methods siRNA targeting MK was used to downregulate MK expres-sion in human breast cancer cell line MDA-MB-231. Then, the experiment was divided into three groups: the untreated group, the empty vector transfection group and MK gene interference group. CCK-8 assay was used to detect the endothelial cells proliferation, tr-answell method was for detection of endothelial cell migration numbers, and matrigel in vitro small tube formation assay was used to sur-vey the state of tube formation. Expression of MK in siRNA transfection was identified by RT-PCR and Western blot. Results The MK gene interference group showed lower cell proliferation activity, less number of migration of cells and tube formation than the other two groups (P<0. 05). Conclusion Low-expressed MK in human breast cancer cell line MDA-MB-231 can inhibit proliferation, mi-gration and angiogenesis of endothelial cells, which shows that MK may play an important role in tumor angiogenesis.

Objective To investigate the feasibility and clinical effect for percutaneous microwave ablation of type I substernal goiter under the guidance of ultrasound . Methods Thirteen patients with type I substernal goiter were selected . All patients underwent percutaneous microwave ablation treatment under the guidance of ultrasound . Thirteen patients with 47 medals nodules were performed ultrasound guided percutaneous microwave ablation . Intraoperative heat blocking blood flow was used to prevent severe hemorrhage .Liquid isolation belt and leverage from methods were used to effectively prevent surrounding important structures against heat damage .Small amount of residual method was used to avoid tracheal collapse . All patients were followed up post‐operative 1 ,3 ,6 ,12 months by monitoring of thyroid nodule volume change , adverse reactions , and postoperative complications . Results Thirteen patients with 47 medals nodules were successfully performed percutaneous microwave ablation guided by ultrasound . Among them ,2 patients performed the second treatment for too large volume of goiter ,tracheal displacement and poor physical tolerance of older . The postoperative ultrasound contrast and color Doppler showed that the lesions were completely inactivated . No obvious complications occurred in all 13 patients , and no permanent hoarseness occured . The symptoms of cervical oppression and discomfort disappeared for all patients within 1-4 months after surgery . Thirteen patients were followed up for 12 months after the operation . The thyroid function was normal and the volume reduction rate of thyroid nodules was (85 ± 31)% . Conclusions Percutaneous microwave ablation of type I substernal goiter under ultrasonic guidance is a safe and effective method to reduce the thyroid nodules with no serious complications . It is worth to be popularized in clinical practice .

Objectives To investigate the effects of sodium arsenite (NaAsO2) on the expression ot microRNA-191 (miR-191) and tissue inhibitor of metalloproteinase 3 (TIMP-3) in human normal hepatic cells (L-02 cells).Methods L-02 cells were exposed to different doses of NaAsO2 [0 (control group),5,25,50 and 75 μmol/L]for 24 h,or treated with 5 and 25 μmol/L NaAs02 for 0 (control group),12,24 and 48 h.The miR-191 inhibitor was used to suppress the expression of miR-191.qRT-PCR was performed to detect the expression level of miR-191 and TIMP-3 mRNA,and the protein level of TIMP-3 was analyzed by Western blotting.Results Dose-effect study:There were significant differences in the expressions of miR-191,TIMP-3 mRNA and protein between the 5 groups (F =85.674,20.952,123.393,all P ＜ 0.05).The expressions of miR-191 in all groups (1.702 ± 0.124,2.077 ±0.234,2.145 ± 0.105,2.003 ± 0.077) were higher than that of control group (0.990 ± 0.035,all P ＜ 0.05);the mRNA expressions of TIMP-3 in 25,50,75 μmol/L groups (0.848 ± 0.067,0.804 ± 0.081,0.813 ± 0.076) were all lower than that of control group (0.996 ± 0.007,all P ＜ 0.05),but there was no significant difference in the mRNA expression of TIMP-3 between the 5 μmol/L group and control group (0.939 ± 0.133 vs 0.996 ± 0.007,P＞ 0.05),and the protein expressions of TIMP-3 in all groups (0.846 ± 0.093,0.611 ± 0.123,0.554 ± 0.098,0.529 ± 0.067) were lower than that of control group (1.006 ± 0.003,all P ＜ 0.05).Time-effect study:there were significant differences in the expressions of miR-191,TIMP-3 mRNA and protein between the exposure groups of 5 and 25 μmol/L (For 5 μmol/L:F =86.355,16.404,22.898,all P ＜ 0.05;For 25 μmol/L:F =104.321,20.123,52.321,all P ＜ 0.05).The expressions of miR-191 in all exposure groups of 5 and 25 μmol/L (1.392 ± 0.152,1.691 ± 0.167,2.018 ± 0.130 and 1.456 ± 0.167,1.946 ± 0.178,2.259 ± 0.256) were higher than those of control groups (1.001 ± 0.014,1.008 ±0.027,all P ＜ 0.05);the mRNA expressions of TIMP-3 in 48 h exposure group of 5 μmol/L and all exposure groups of 25 μmol/L (0.824 ± 0.093 and 0.897 ± 0.033,0.815 ± 0.089,0.709 ± 0.103) were lower than those of control groups (1.004 ± 0.018,0.997 ± 0.057,all P ＜ 0.05),but there were no significant differences in the mRNA expressions of TIMP-3 between the 12,24 h exposure groups of 5 μmol/L and control group (0.952 ± 0.072,0.929 ± 0.121 vs1.004 ± 0.018,all P ＞ 0.05);the protein expressions of TIMP-3 in all exposure groups of 5 and 25 μmol/L (0.857 ±0.068,0.832 ± 0.106,0.691 ± 0.112 and 0.785 ± 0.097,0.620 ± 0.066,0.453 ± 0.075) were lower than those of control groups (1.006 ± 0.045,1.004 ± 0.078,all P ＜ 0.05).The treatment of miR-191 inhibitor:there were significant differences in the expressions of miR-191 and TIMP-3 protein between different groups (F =104.306,67.015,all P ＜ 0.05).The elevated expression level of miR-191 induced by NaAsO2 was significantly suppressed after transfected with miR-191 inhibitor (0.314 ± 0.094 vs 2.051 ± 0.371,P ＜ 0.05),which in turn up-regulated the protein expression of TIMP-3 (1.965 ± 0.277 vs 0.541 ± 0.183,P ＜ 0.05).Conclusion The expression level of miR-191 is elevated in response to NaAsO2 exposure,and miR-191 has subsequently suppressed the expression of TIMP-3,a potential target of miR-191.

Objective To compare treatment outcomes and antibiotic related costs of A. baumannii infection treated with imipenem cilastatin and ampicillin sulbactam and to provide evidence for clinical treatment of A. baumannii infection. Methods A prospective investigation was performed on 71 ICU A. baumannii infection patients treated with imipenem cilastatin and ampicillin sulbactam. The treatment outcomes and antibiotic related costs were also analyzed. Results There was no significant difference between the therapeutic efficacy of these two drugs within the same time limit. The antibiotic related costs of the patients treated with ampicillin sulbactam were significantly lower than those with imipenem cilastatin (2 113 Yuan vs 5 789 Yuan , P

ObjectiveTo assess the psychosomatic health status of patients with facial steroiddependent dermatitis by using Pittsburgh sleep quality index (PSQI),self-rating anxiety scale (SAS) and selfrating depression scale(SDS).MethodsPsychosomatic health status was evaluated by using PSQI,SDS and SAS in 92 patients with facial steroid-dependent dermatitis and 50 healthy controls.ResultsThe scores on depression and anxiety in patients with steroid-dependent dermatitis were higher than those in the controls (63.11 ± 10.29 vs.36.78 ± 2.46,53.24 ± 9.98 vs.31.29 ± 9.45,both P ＜ 0.01).There were significant differences between the two groups in the quality of sleep.The general and mean PSQI global scores on subjective daytime function,sleep disturbances,time needed to fall asleep and sleep duration were significantly higher in the patients than in the controls.ConclusionsPatients with steroid-dependent dermatitis have a poor sleep quality with the presence of anxiety and depresssion.Psychosocial interventions are warranted for the treatment of patients with steroid-dependent dermatitis.

Objective To investigate the effect of virtual simulation teaching method in the comprehensive experimental teaching.Methods In experimental teaching of nursing,simulation medical records were compiled,plans were implemented,experiment guidance materials were preliminarily formed and teaching effect was evaluated.Results The application of virtual simulation teaching in nursing comprehensive experiments reflected the comprehensiveness of teaching content,and the comprehensiveness of knowledge,ability and capacity.It could train students creative ability,exploring spirit,enhance the sense of professional and humane care consciousness,train the communication skills and strain capacity,make active teaching atmosphere,improve the quality of teaching.Conclusions This teaching method can make students achieve mastery through a comprehensive multi-disciplinary knowledge,improve the ability of solving practical problems,help nurses to develop the comprehensive professional quality.