Objective:To construct a recombinant lentivirus plasmid of RNA interference targeting (MSLN) gene and to observe its effect on MSLN expression in human ovarian cancer cell line OVCAR-3 and its effect on cell proliferation. Methods: According to the Genbank information of MSLN, four RNA interfering sequences and a negative sequence were designed and inserted into plasmid pRNAT-U6.2/Lenti and 5 kinds of plasmids were packaged: LV-MSLN-negative,LV-MSLN-shRNA1, LV-MSLN-shRNA2, LV-MSLN-shRNA3, and LV-MSLN-shRNA4; and they were used to transfect OVCAR-3 cells. Western blotting and indirect immunofluorescence were then used to investigate the interfering efficiency. The plasmid with high interfering efficiency was packaged. The cell proliferation test and clone-forming test was used to assess the changes in cell proliferation. Results: DNA sequencing showed that the sequences of 5 recombinant lentivirus plasmids were correct. Lentivirus packaging was successfully done. Western blotting analysis confirmed that LV-MSLN-shRNA4 had the highest interfering efficiency (90%). MSLN specifically bound to cytomembrane of OVCAR-3 cells. Expression of MSLN in the interfered cells (OVC-shRNA) was weaker than that in the control cells (OVC-neg,OVC). OVC-shRNA cells([11.2?1.3]?105) grew slowly compared to OVC-neg cells([20.5?2.5]?105) and OVC cells([21.9?2.3]?105) (P

Objective To investigate the function of mesothelin by analyzing the location,the expressions and the adhesive ability of mesothelin and CA125 in human epithelial ovarian cancer.Methods Mesothelin and CA125 protein expressions and locations in human epithelial ovarian cancer tissues and cell lines were determined by indirect immunofluorescence double maker and Western blot.Cell adhesive ability was detected by EDTA-induced cell detachment.Results Mesothelin and CA125 were specifically located in cytomembrane of human epithelial ovarian cancer cells.Expressions of mesothelin in the SKOV-3 and 3AO cells were weaker than those in the OVCAR-3.SKOV-3 and 3AO cells adhered less effectively to plastic substance and matrigel than OVCAR-3(P

Objective To study the DNA damage and repair of normal lung interstitial cells and human lung adenocarcinoma cells exposed to cigarette smoke. Methods Cultured human embryo lung fibroblasts (HLF) and human lung adenocarcinoma A549 cells. Mainstream smoke was collected by using dimerhyl sulfoxide (DMSO) and phosphate buffer solution (PBS) as absorbents. MTT assay was used to test the cytotoxicity of the solutions of cigarette smoke, then selected the concentrations of the solutions with no obvious cytotoxicity to treat cells and detected DNA damage and repair by comet assay. Results As treated with original solutions or 1/2 dilution of DMSO cigarette smoke solutions only, the Viability of cells was below 80%, but it was beyond 80% when treated with PBS solutions. The results showed that a significant difference of DNA damage was seen between the treated groups and negative control groups (P0.05),but the DNA damage caused by DMSO solutions was worse than PBS solutions significantly (P

OBJECTIVE To investigate the protective mechanism of recombinant bactericidal permeability-increasing protein 21(rBPI21) in rat endotoxemia.METHODS The different dosage of rBPI21 in endotoxemia rats was injected and the changes in lipopolysaccharide(LPS), lipopolysaccharide binding protein(LBP) and tumor necrosis factor ?(TNF?) contents in blood of different group rats were continuously observed.RESULTS At 6 and 12 hours,the levels of LPS in rBPI21 treatment 1 group(rBPI21 dosage 0.625 mg/kg) were significantly lower than those in endotoxin group at the same time.Serum LBP and TNF? in rBPI21 treatment 1 group were both lower than those in endotoxin group at any time point.Compared with the endotoxin group,the levels of LPS,LBP,and TNF? in rBPI21 treatment 2,3 and 4 groups(rBPI21 dosage 1.25 mg/kg,2.5mg/kg,and 5.0mg/kg,respectively) markedly dropped at any time points,the survival rates were increased from 16.7%(endotoxin group) to 58.3%,91.7% and 100%(rBPI21 treatment 2,3,and 4 groups) individually.CONCLUSIONS The protection of rBPI21 in endotoxemia rats is primarily achieved through neutralizing LPS,decreasing LPS activity in vivo and inhibiting LBP and TNF-? synthesis.

Objective To analysis the consequence of current perception threshold (CPT) by different frequency and time of transcutaneous electrical nerve stimulation (TENS). Methods CPT of foramen area and arm area was measured to evaluate the effect of TENS. Different frequency and time of TENS was given to 30 healthy volunteers. Stimulating to Hegu, CPT of the foramen area and the arm area on the same side was measured. Results CPT of the foramen area increased with stimulation. Low frequency of TENS inhibited the chronic pain significantly(P＜0.05),high frequency of TENS inhibited both of the chronic pain and the acute pain(P＜0.05). The inhibition of pain is more influenced by the frequency of TENS than the time of it. Conclusion Channel and point of TENS can influence CPT of the specific reaction area. And the choice of appropriate frequency of the treatment is more important than extending the treatment time alone.

Objective To investigate the effect of transitional care of physical and mental health of the thoracic spinal cord injury and paraplegic patients discharged on its recovery. Methods Randomized controlled analysis of patients in Spine and Spinal Cord Injury Surgery between June 2014 to October 2015, a total of 65 patients with thoracic spinal cord injury and paraplegic were studied, according to hospital mantissa odd and even numbers divided into observation group (35 cases) and control group (30 cases). The observation group was instituted to extending posthospital care, not the control group. The self-care ability and psychological state were evaluated by Function Independent Measure (FIM), Hamilton Anxiety Scale(HAMA), Hamilton Depression Scale (HAMD)within discharge and in 3 months. And the incidence of complications between the two groups were compared. Results At discharge, the difference of FIM, HAMA, HAMD score between the observation group and the control group was not statistically significant (P>0.05). Discharged after 3 months, the score of FIM, HAMA, HAMD in observation group was(72.35 ± 4.35),(4.80 ± 4.50),(12.13 ± 7.25)points, (63.95 ± 5.02),(10.15 ± 4.91), (19.48 ± 8.31)points in control group. The difference between the two groups was statistically significant (t=-4.798, 3.524, 3.110, P<0.01). There was no significant difference in the incidence of pulmonary infection and blood clots between two groups (P > 0.05); The incidence of urinary tract infection, pressure ulcers, constipation were 8.57% (3/35), 5.71% (2/35), 17.14% (6/35) in observation group, 40.00% (12/30), 33.33%(10/30), 63.33%(19/30) in control group, there were significant differences between, two groups (χ2= 4.324, 4.219, 5.337, P < 0.05). Conclusions Transitional care can significantly improve the psychological state of patients with thoracic spinal cord injury and paraplegia discharged, reduce the incidence of complications and improve quality of life.

Objective To analyze the mechanism of pigment epithelium?derived factor(PEDF)regulation in lung cancer cell proliferation and migration by vascular endothelial growth factor(VEGF). Methods VEGF expression was observed after overexpression or silencing of PEDF. Proliferation and migration were analyzed by MTT and transwell assays. Real?time PCR and Western blotting were performed to investigate the mechanism underlying PEDF regulation of the VEGF/SRC/FAK pathways. Results PEDF could inhibit the proliferation and migration of A549 cells by VEGF. Conclusion PEDF can be considered as a potential therapeutic target for lung cancer.

Objective To explore the effect of evidence-based nursing on improving the compliance of chronic obstructive pulmonary disease (COPD) patients with noninvasive positive pressure ventilation. Methods Forty-five patients hospitalized in the first half of the year were assigned as control group and another 45 patients hospitalized in the second half of the year were as experiment group. The control group was treated with traditional nursing methods. In the experiment group, a nursing panel was set up to analyze the causes of noncompliance of COPD patients with noninvasive positive pressure ventilation and gave care to the patients based on the analyses. The two groups were compared in terms of adverse reactions and the compliance with treatment. Result The rates of compliance with treatment and adverse reactions such as fear and anxiety , the man-machine confrontation and flatulence in the experiment group were significantly lower than those of the control group obviously (P<0.05). Conclusion The evidence-based nursing can reduce the incidence of adverse reactions , and enhance the compliance of COPD patients with noninvasive positive pressure ventilation effectively.

Objective:To study the effect of different doses of oxycodone on the serum thromboxane A2 (TXA2),plasma endothelin (ET) levels and immune function of patients underwent laparoscopic cholecystectomy.Methods:90 patients of elective laparoscopic cholecystectomy who were treated from August 2013 to August 2016 in our hospital were selected and divided into 3 groups by random number table,with 30 cases in each group.At the beginning of operation,they were given intravenous oxycodone,group A was given 0.1 mg/kg oxycodone,group B was given 0.2 mg/kg oxycodone,group C was given 0.3 mg/kg oxycodone.The changes of hemodynamics,serum TXA2 and ET levels were compared between the three groups at T0 (after admission),T1 (after anesthesia induction),T2 (after intubation),T3 (gallbladder separation),T4 (end of surgery) and the changes of immune function was compared at T0,T5 (postoperative 2h),T6(postoperative 1d),T7 (postoperative 3d);and the extubation time,recovery time,hypotensor,additional analgesics situationin and adverse reactions were recorded.Results:The diastolic pressure (DBP),systolic blood pressure (SBP),heart rate (HR) in three groups at T2,T3 point was significantly higher than T0 point(P＜0.05),the DBP,SBP and HR in the B,C groups were significantly lower than the A group at T2 and T3 point(P＜0.05);the TXA2 in three groups at T2,T3,T4 point was significantly higher than T0 point(P＜0.05),and A group＞B group＞C group,there was significant difference between the two groups (P＜0.05);the ET in three groups at T2,T3 point was significantly higher than T0 point(P＜0.05),the ET in the B,C group was significantly higher than the A group at T2,T3 point(P＜ 0.05);the CD3+,CD4+,CD8+ and CD4+/CD8+ of the three groups at T5 point were significantly lower than that ofT0 point (P＜0.05),the CD3+,CD4+,CD8+ and CD4+/CD8+ in A group were significantly lower than that of B,C group at T5 point (P＜0.05);the extubation time in C group was significantly longer than that of A,B group(P＜0.05);the total incidence of adverse reactions in C group was significantly higher than that the A,B group (P＜0.05).Conclusion:In the laparoscopic cholecystectomy,application of 0.2 mg/kg oxycodone had little effect on hemodynamics,serum TXA2,ET levels and immune function with higher safety.

Aim To prepare soluble human C1 q and tumor necrosis factor related protein-6 in Escherichia coli and analyze the bioactivity. Methods Recombi-nant plasmid was transformed into E. coli expression strain, and the recombinant protein Trx-hCTRP6 was expressed induced by IPTG and then purified. Results Trx-hCTRP6 was expressed efficiently and purified using Ni-NTA affinity chromatography and Superdex G-75 column. The purified Trx-hCTRP6 was shown to be active under in vivo and in vitro assay conditions. Con-clusion Active Trx-hCTRP6 is efficiently prepared from E. coli protein expression system.