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 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 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.

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

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.

A total of 1342 individuals underwent physical examinations according to the criteria of metabolic syndrome of International Diabetes Federation (IDF) in 2005.And 314 patients with metabolic disorders were screened for diabetes by standard meal and methods oral glucose tolerance test (OGTT).Newly diagnosed diabetics was 12 (4.1％ ) vs.17 (5.8％) respectively.No significant difference existed between two methods (P =0.332).Kendall's (τ)b =0.313,Kendall's (τ)c =0.208 and Gamma coefficient =0.580 (P =0.000).The mixed meal method was correlated with OGTT,Kappa =0.258 (P =0.000) and two methods were consistent.Diabetic screening should be stressed in the subjects with metabolic abnormalities.And the detecting efficiency of postprandial plasma glucose is similar to OGTT.

Objective:To highlight the key points of primary culture of human nasal epithelial cells by enzymatical dissociation for high achievement ratio,and to establish a successful primary culture model for subsequent experiments.Method:Primary culture of human nasal epithelial cells was performed with enzymatical dissociation of isolated tissue in serum-free medium. On the basis of this method,some improvements were subjected,such as stripping mucosal epithelium from adjacent connective tissue,applying DNase type Ⅰ to digesting procedure,adding trypsin directly to the collagenase solution containing digested mucosa pieces,employing uncoated culture dishes and so on. Immunofluorescence with a monoclonal anti-cytokeratin antibody 8/18 was used to confirm the epithelial nature of the cultured cells. Result:Nasal epithelial cells grew well and confluenced on the 6th to 8th day. Positive expression of cytokeratin(CK)8/18 showed the epithelial property of cultured cells.Conclusion:Primary culture model of human nasal epithelial cells can be successfully established by enzymatical dissociation. Improvements on processes of material using and enzyme digestion can gain a high achievement ratio and harvest a high purity and certain amount of reliable primary epithelial cells.