AIM: The current study was designed to construct eukaryotic expression vector containing NK4 gene and transfect it into human pancreatic cancer cell lines.METHODS: The recombinant of pcDNA3/hNK4 was digested by restriction enzyme, the NK4 gene was cloned into a high effective eukaryotic expressing plasmid which contains CMV2 immediate early gene promoter and then transiently introduced into the pancreatic cancer cell line SW1990 by lipofectamine and clonal cell lines that secrete high levels of NK4 protein were isolated.The expression of NK4 was observed by RT-PCR and Western blot, in vitro the vascular endothelial cell proliferation inhibiting activity of NK4 was examined by 3-[4,5-dimethylthiazolzyl]-2,5-diphenyl tetrazolium bromide(MTT) method. RESULTS: A specific expression of NK4 gene mRNA by lipofectamine-mediated transfer exhibited only in SW1990/NK4 cells,Western blot analysis demonstrated that there was positive expression of NK4 protein(50 kD).The NK4 inhibited proliferation of the vascular endothelial cells in vitro. CONCLUSION: The recombinant of pRC/CMV2-hNK4 is a high effective expressing eukaryotic vector.The bio-engineering product of the NK4 is an angiogenesis inhibitor and may play an important role in the gene therapy for tumor.

0.05);the annual recurrence rates of hemorrhage of groupⅠand groupⅡ were 35.0% and 15.0% respectively,the recurrence rates of hemorrhage in group Ⅱ was significantly lower than that of group Ⅰ(P

Objective To study the clinical features and protective measures of AIDS with fungal esophagitis as the initial presentation,improve the knowledge of non-specialist medical staff of AIDS. Methods The clinical manifestations,laboratory tess.endoscopic examination,and medical protective measures of 15 AIDS patients with fungal esophagitis were analyzed. Reults The patients were infected through sexual transmission mainly,and manifestation included difficuhy in swallowing,retrostemal pain,weight loss,fever,loss of appetite,fatigue and other symptoms.The patients got confirmed diagnosis by our hospital after visiting many other hospitals.3 patients were automatically discharged after confirmed diagnosis,other patients impreved after antifimgal therapy and symptomatic treatment.No iatrogenic infection occurred during hospitalization. Conclusions Non-specialist medical staff in AIDS should recognize the clinical features of AIDS,give timely confirmed diagnosis and treatment,avoid missed diagnosis and misdiagnosis,strengthen protective measures and avoid iatrogenic infection.

Objective To investigate the systemic effects of Helicobacter pylori (Hp) infection by comparing differential counts of leukocytes in peripheral blood and C-reactive protein (CRP) levels before and after eradication of Hp. Method A total of 158 Hp-positive patients underwent eradication by standard one-week triple therapy were divided into Hp eradicated group(n = 108) and Hp non-eradicated group(n = 50),the populations of peripheral blood leukocytes and CRP levels before and 0 (just after therapy),1 and 12 months after eradication were retrospectively analysed. Results In the eradicated group,blood leukecytes,neutrophils and monocytes decreased significantly after eradication,and the CRP levels were significantly lower than the levels before treatment (P < 0.05 ),but there was no significant change in eosinophils,basophils and lymphecytes (P > 0.05).In the non-eradicated group,there was no significant change in any studied pa-rameter (P > 0.05). Conclusion These findings suggest that Hp infection increase blood leukocytes,neu-trophil and monocyte counts in the peripheral blood and CRP levels,which indicates a significant role of Hp infection in systemic disorders.

Objective To explore and analyze the pathogen proifle and risk factors of infected diabetic foot ulcers, and propose appropriate prevention and intervention measures for early recovery of patients.Methods The data of 120 patients with diabetic foot ulcers treated in our hospital from February 2012 to May 2014 were retrospectively analyzed. These patients were classified into infection group (38 patients) and non-infection group (82 patients) based on whether their ulcer was infected or not. The pathogens and risk factors of infection in diabetic foot ulcer were analyzed.Results In this study, 43 strains of pathogenic bacteria were isolated from 38 cases of infection, of which gram positive bacteria accounted for 51.2 %, gram negative bacteria 41.9 %, and fungi 7.0 %.Staphylococcus aureus,Pseudomonas aeruginosa and Enterobacter cloacae were the top three pathogens, accounting for 16.3 % each. Univariate analysis showed that diabetic vascular complication, retinopathy, more than 2 foci of ulcer, osteomyelitis, and diabetic nephropathy were closely associated with infection (P<0.05). Multivariate logistic regression analysis showed that HDL-C, albumin, diabetic vascular complication, number of ulcers, osteomyelitis were independent risk factors for infection in patients with diabetic foot ulcer. Conclusions HDL-C, red blood cell, hemoglobin, diabetic complications, white blood cell, triglyceride, albumin, diabetic vascular complication and smoking are the risk factors for infection in patients with diabetic foot ulcer. These factors should be prevented and controlled to reduce the incidence of infection in diabetic foot ulcer.

Objective To investigate the clinical effect of the treatment of senile chronic superficial gastritis with the treatment of the elderly patients.Methods Collected in our hospital diagnosed by endoscopy in 80 elderly patients with chronic superficial gastritis were randomly divided into control group and treatment group, 40 cases in each group, the control group using ranitidine therapy, treatment group with teprenone treatment, evaluation of 2 groups of drugs on chronic superficial gastritis and therapeutic effect safety.Results The total effective rate of treatment group was 92.5%, control group total effective rate was 75%;the treatment of two patients with total efficiency, the total effective rate in treatment group was significantly higher than the control group(x2=4.50, P<0.05);the average time for improvement of symptoms in the treatment group was(8.32 ±2.06)days, the average time to improve the symptoms of the control group(11.69 ± 2.12)days, there was significant difference between the two groups(t=7.21, P<0.05), the treatment group the incidence of adverse reaction was 5%, the control group adverse reaction rate was 10%, the difference between the two groups was not statistically significant(x2=0.18, P>0.05).Conclusion It is safe and effective for the treatment of chronic superficial gastritis in elderly patients.

Objective To develop a clinically useful assay for detecting the mutations of HBV pre-C/BCP based on the pyrosequencing and accuracy, reproducibility and reliability of this assay was evaluated. Methods The pyrosequencing primers for HBV pre-C/BCP mutation were designed through the cluster analysis among one hundred HBV gene sequences. After the amplification of the fragment of pre-C/BCP with the template of pre-C/BCP mutation plasmids, the pyrosequencing method for pre-C/BCP detection was initially set up with this standard sample. The accuracy, reliability and reproducibility of the pre-C/BCP pyrosequencing were confirmed through the pre-C/BCP plasmids as a standard sample when compared with Sanger/genechip sequencing method pre-C/BCP pyrosequencing assay was applied for detecting pre-C/BCP mutation types of 60 chnical serum samples in HBV patients. Results The pre-C/BCP mutation detection assay based on pyrosequencing has been established in our study. The coincidence rate between pyrosequencing and Sanger squencing was 100%. The coincidence rate between the result of pyrosequencing and of genechip method was 91.7%. The reproducibility of this assay was 97. 8%. It indicates the pre-C/BCP pyrosequencing is a high-accurate method with, good-reproducibility and high-reliability. And multi-site detection can be achieved by pyrosequencing one time. A rare mutation T1758C was also detected. Conclusion Pyrosequencing for pre-C/BCP mutations assay is high-throughout method for simultaneous detection of multi-site mutation.