Objective:To investigate the mechanism of WWOX regulating immunosuppression in Lewis lung cancer cells.Methods:Transfected pCDNA4.0/myc-WWOX plasmid was transfected into Lewis lung cancer cells to detected the effect of cell supernatant on generation of lymphocytes by MTT method,and mRNA expression of immunosuppression factors by semi-quantitative RT-PCR.Function change of NK,CTL to kill tumor and lymphocte proliferation were analyzed by MTT and tumor volume.Results:The suppression of the supernatant to generation of lymphocytes was present compared with control group(P

Objective To investigate the relationship between pluse pressure(PP) and serum levels of C-reactive protien(Hs-CRP) in elderly patients with HBP.Methods To measure serum levels of Hs-CRP,total cholesterol(TC),triglyceride(TG),high density lipoprotein-cholesterol( HDL-C),low density lipoprotein-cholesterol(LDL-C) and FBG in 160 patients. Patients were divided into four groups according to the levels of their PP,100mmHg whose serum levels of Hs-CRP were compared.Results Hs-CRP rose with the levels of PP,and were significantly higher in four groups(P

Objective To evaluate the changes of alveolar cells apoptosis and the expression of related genes in pulmonary injury induced by ischemia/reperfusion (I/R) in acute experimental pulmonary embolism of rabbits. Methods The left lung artery of rabbits were obstructed by inflating gas of 5F Berman sacculus catheter, then the gas of sacculus was put out to result in blood reperfusion. Thirty-six New Zealand rabbits of either gender and weighing 2.5 to 3.0 kg were studied, and randomly divided into six groups: control group (control), sham operation group (sham), ischemia 1h group (I 1h), ischemia 2h group (I 2h), ischemia2h and reperfusion 1h group (IR 1h), ischemia 2h and reperfusion 2 h group (IR 2h). Alveolar cells apoptosis and Bax, Bcl-2, Fas/FasL protein expression were studied By using flow-cytometry and immunocytochemistry techniques. Results The lung apoptosis cell number was increased in ischemia groups compared with control and sham group. The reperfusion after ischemia further increased the apoptosis cell numbers compared with simple ischemia, and the longer time reperfusion,the larger number of cell apoptosis. Bax, Bcl-2, Fas and FasL protein expression were significantly higher in ischemia, ischemia/reperfusion group than that in those of control groups (P

Objective To construct eukaryotic expressing vector of human melanin-concentrating hormone receptor 1(MCHR1), then to transfect CHO cells with the vector for establishment of stable CHO cell line. Methods The full-length MCHR1 cDNA fragment was amplified by PCR from the human fetal brain cDNA library and then inserted into eukaryotic expression vector pcDNA3.1(+).The recombinant was transfected into CHO cells by lipofectamine TM 2000 after identification of digestion and sequencing on the recombinant eukaryotic expression vector pcDNA3.1(+)/MCHR1. The stable transfected CHO cell line was then established by screening cultures with G418, and the transcription and expression of MCHR1 were identified by RT-PCR, Western blot and immunofluorescence. Results The eukaryotic expression vector pcDNA3.1(+)/MCHR1 was constructed successfully, stable transfected CHO cell line was established, the MCHR1 protein was expressed successfully. Conclusion The construction of eukaryotic expression vector pcDNA3.1(+)/MCHR1 and the establishment of stable transfected CHO cell line provided a solid experimental foundation for further studies on the function of MCHR1.

Objective To construct eukaryotic expression vectors expressing short hairpin RNA(shRNA)sections targeting human MCHR2 and to observe their effects on MCHR2 gene expression in HEK293 cell line.Methods According to the sequence of human MCHR2 gene,the oligonucleotides of shRNA were designed and synthesized and directionally cloned into plasmid pGenesil-1 with enhancing green fluorescence protein(EGFP)gene and Kan gene.The recombinant vectors were confirmed by enzyme digestion analysis and DNA sequencing.The recombinant vectors were transfected into HEK293 cell line by LipofectamineTM2000,the effects on MCHR2 at mRNA and protein levels were observed.Results Four shRNA expressing recombinants and the corresponding negative control vector were constructed and transfected into HEK293 cell successfully.MCHR2 transcript was reduced by about 45.8%-66.4%,the protein of MCHR2 was reduced by about 44.2%-81.0% in four transfectants respectively.Conclusion The construction of eukaryotic expression vectors expressing shRNA sections targeting human MCHR2 and identification successfully established a favourable foundation for further study on the function of MCHR2.

BACKGROUND: What role tumor-related genes play in the process of tumor generation, development, metastasis and prognosis has always been a thorny issue in medical field?OBJECTIVE: To study the detection of gene mutation in tumor by denaturing gradient gel electrophoresis(DGGE) and automated DNA se+uence analysis and the change of p53 gene and p53 protein during the development and metastasis of colorectal carcinoma so as to provide basis for evaluating the prognosis of colorectal carcinoma.DESIGN: Single sample study using the tissue specimen as subject.SETTING: Department of oncology in an affiliated hospital of a military medical university.PARTICIPANTS: We collected the primary focus and liver metastasis focus specimens from 41 patients with colon cancer who had hepatectomy because of liver metastasis 5 months to 5 years after radical operation for coloncancer. They were inpatients in Nanfang Hospital, First Military Medical University of Chinese PLA, from January 1994 to December 2000.METHODS: p53 gene(exons 5- 11) mutation of primary focus and liver metastasis focus specimens from 41 cases of colon cancer was examined by DGGE and automated DNA sequencing. Expression of p53 protein was detected by immunohistochemistry.histochemical staining.MAIN OUTCOME MEASURES:①Detection of the mutation of p53 gene by DGGE;②Analysis of p53 gene sequence;③Results of p53 immunohistochemical staining.RESULTS: p53 gene mutation was detected in exons 5 - 9 in 24 out of 41patients(62% ) . Among them, 6 patients had p53 mutation in liver metastasis. The others had consistent mutations in both primary coloreetal and hepatic metastatic lesions. In addition, p53 mutation was also found in the metastatic lesion in three patients. Among the 16 cases of mutation in primary colorectal and hepatic metastatic lesions, 14 cases showed that the ratio of p53 base peak to normal peak was significantly higher in hepatic metastatic lesions than in primary colorectal lesions(P ＜ 0. 001) . Results of p53 immunohistochemical staining were highly consistent with those of DGGE and DNA sequence analysis. However, gene analysis detected focus with nonsense mutation while immunohistochemistry detected overexpression of p53 protein.CONCLUSION: p53 mutation, in patients with colorectal carcinoma followed by hepatic metastases, mostly originates from primary colorectal lesion and then is kept and metastasizes into hepatic cells. The amount of mutated p53 gene and the number of tumor cells containing p53 mutation are increased in hepatic metastatic lesion. P53 mutation is positively correlated with overexpression of p53 protein.

Objective To measure the bone mineral density in children with dual-energy X-ray absorptiometry and examine the relevance of bone mineral density with the risk of pediatric upper limb fracture.Methods A total of 864 children (8-14 years of age) admitted into our hospital in 2009 were involved in the study.The bone mass density (Z score) at the left radius was measured using dual-energy X-ray absorptiometry.Children were grouped according to their Z score and age to observe correlations of Z value and age with the incidence of fractures.Results Thirty-three children were lost to follow-up.For the other 831 children,the mean duration of follow-up was 2 years.Z sore was ≤-2 in 66 children and ＞-2 in 765 children.There was significant difference in fracture incidence between children with Z score of ≤-2 and ＞-2 (27.27％ vs 5.49％,P ＜0.01).Furthermore,fracture incidence in children at age group of 8-10 years and 11-14 years revealed significant difference (10.83％ vs 5.03％,P ＜ 0.01).Conclusions Bone mineral density is closely related to upper limb fracture in children and increases with aging.Early application of dual-energy X-ray absorptiometry to detect the bone density is an effective primary measure for prevention of fractures in children.

In this article, HPLC fingerprint analysis method for total flavonoids from pollen of Brassica campestris L.was established., The HPLC fingerprint was performed on Waters C18 column (250 mm × 4.6 mm, 5 μm), eluted gradiently with the mixture of acetonitrile and 0.4% phosphoric acid aqueous solution at a flow rate of 0.8 mL·min-1. The column temperature was 40℃. The detection wavelength was 320 nm. The HPLC standard fingerprint of total flavonoids from pollen of Brassica campestrisL. was established, and 16 common peaks were calibrated. The method was simple, stable, and reproducible. It could be applied for quality control of total flavonoids from pollen of Brassica campestris L.

BACKGROUND:Pain is the significant cause for patients with early rehabilitation after total knee arthroplasty. Continuous epidural analgesia and continuous femoral nerve block are effective analgesic methods after total knee arthroplasty, however, which method has better effects and less complications remains controversial. OBJECTIVE:To compare the efficacy and safety of countious femoral nerve block and continuous epidural analgesia after total knee arthroplasty. METHODS:We searched Cochrane Library, PubMed, EMBASE, Web of Science, CBM, CNKI, VIP, and WangFang. Meanwhile, we also searched conference papers and academic dissertation. The retrieval time was from database establishment to October 1, 2014. Studies of randomized control ed trials on countious femoral nerve block and continuous epidural analgesia after total knee arthroplasty were included. We evaluated the quality of these included studies and analyzed data by Cochrane Col aboration’s RevMan 5.0 software. RESULTS AND CONCLUSION:A total of 12 randomized control ed trials (4 English articles and 8 Chinese articles) involving 680 patients were included. There were 343 patients with countious femoral nerve block and 337 patients with continuous epidural analgesia. Meta-analysis results revealed that no significant differences in visual analog scale scores were detected between the countious femoral nerve block and continuous epidural analgesia groups at 6, 12, 24 and 48 hours after total knee arthroplasty. However, compared with the continuous epidural analgesia group, countious femoral nerve block could decrease the incidences of nausea/vomiting (RR=0.36, 95%CI:0.21-0.63, P=0.003), urine retention (RR=0.08, 95%CI:0.04-0.16, P<0.001) and dizziness (RR=0.24, 95%CI:0.06-0.99, P=0.05). These results indicate that compared with epidural analgesia, countious femoral nerve block after total knee arthroplasty provided a strong analgesia effect, contributed to early functional training, had less adverse reactions, and was a safe and effective analgesic method.

Objective By reviewing the relevant literature, we explored the current status and problems of the evaluation of the training effectiveness, and put forward the corresponding countermeasures, in order to provide a theoretical basis for the effect evaluation of nurse training in China. Methods By searching Citation online search and integration system (CITE), the Chinese biomedical literature service system (CBM), we included the literatures on the effect evaluation of the nurses training in the core journals from January 2006 to November 2015. Retrieval literatures were imported the bibliographic management software Note express by database. The corresponding field were analyzed in the literature′s age distribution, evaluation object, evaluation model, evaluation method, evaluation index and evaluation tool, et al. Results There were 343 literatures had retrieved, the total number of literatures presented a increasing tendency. The evaluation methods, evaluation tools and evaluation indicators presented a tendency of diversity, and the evaluation model was relatively single. Only 16 (4.7%) literatures tried to construct the training effect evaluation system of nurses, which was not based on some theory or hypothesis, which had not been effectively put into practice. Conclusion The effect evaluation system of nurse training needed to be done in depth, such as reconstruction of the new training evaluation model, the establishment of training evaluation system of different professional, different levels, different positions, to research evaluation methods, evaluation criteria and clinical practice testing, to design, training evaluation management software etc.