Objective To construct the eukaryotic expression vector of human PRKCB1 containing enhanced green fluorescence protein gene and transfer into human umbilical vein endothelial cells(HUVECs),then identify activity of expression protein.Methods The pReceiver-M29-PRKCB1 eukaryotic expression plasmid was constructed by frame amplified from pMD18-T-PRKCB1 plasmid.Then the recombinant plasmids were identified by enzyme analysis and DNA sequencing.According to optimized conditions,the eukaryotic expression plasmids were transfered into HUVECs and observed under fluorescence microscope.After that,transfection efficiency was calculated under random vision.The plasma membrane/cytosol ratio of fluorescence was calculated under confocal microscope.The translocation was identified.Results The gene sequence was completely consistent with that reported in GenBank.The enhanced green fluorescence protein was observed in HUVECs after 48 hours.Transfection efficiency was 18.6%?1.6%.The translocation was observed.Conclusion The eukaryotic expression plasmid is successfullyconstructed and transfered into HUVECs,the translocation was identified.It is a potential tool for screening HUVECs stably expressing human protein kinase C ?2 and isolating protein complex.

Objective To observe the influence of long-term smoking on the expression of p53 and K-ras in rat lung tissues, and to study the relationship of smoking to the mutation of p53 and K-ras gene. Methods The model of Wistar rat smoking was built up. Seventy-two healthy male Wistar rats were divided into the experimental group and control group at random. The rats of the experimental group were compelled to smoke, and the rats of the control group were given the same condition as the experimental group was, without smoking. The rats of the experimental group were smoked for 6 months. At the end of each month, 6 rats were chosen from the two groups respectively, their lung tissues were sampled and immunohistochemistry was applied to observe the expression of p53 and K-ras in lung tissues. Finally, the mutation which might happen in the exon 5, 6, 7-8 of p53 and the exon 1 of K-ras was examined by PCR-SSCP. Results The p53 protein was expressed in cell nucleus and K-ras in cytoplasm. The positive ratio of protein expression was increased with the extension of smoking time. The mutation of p53 was increased as the smoking time extended. But the effect of smoking time was not that significant on the mutation of K-ras.Conclusion Smoking can strengthen the expression of p53 and K-ras protein and can also result in gene mutation. As the time of smoking extended, those phenomenons were tending to rise. That provided the theoretical evidence which can be used to judge the lesion of lung tissues caused by smoking and help the early diagnosis of smoking-related lung carcinomas. It is of great theoretical and application values.

Aim To study the effects of probucol on THP-1 macrophage apoptosis and CD36、Caveolin-1 expression induced by ox-LDL.Methods Apoptosis of THP-1 macrophages was determined by flow cytometry analysis.RT-PCR and immunofluorescence were used to detect CD36,Caveolin-1 mRNA level and protein expression respectively.Results Probucol had no effect on mRNA level of CD36,Caveolin-1 in THP-1 macrophages,but it attenuated Caveolin-1 protein expression.Conclusions Probucol can inhibit apoptosis induced by ox-LDL in THP-1 macrophages by down-regulating Caveolin-1 protein expression.

Objective To investigate the effect of expressions of P53 and VEGF in smoking lung cancer and surrounding lung tissue,and to offer us with theorial evidence of early diagnosing smoking-related lung carcinoma clinically.Methods126 lung cancer patients were recruited,including 96 long-term smoking and 30 nonsmoking patients.All clinical data was integrity and the patients had clear smoking history.None of patients underwent chemotherapy,radiotherapy and other tumor treatment.The bronchial epithelium,cancer tissues,pericancer lung tissues,surrounding lung tissues were observed by light microscopy,and the expressions of P53 and VEGF of lung cancer tissues,pericancer lung tissues and surrounding lung tissues were detected by Immunohistochemical methed in smoking and non-smoking group.The experimental data was analyzed by SAS statistical software and the degree of difference between the groups was compared accordingly.ResultsThere were different levels of expansion of alveolar wall in lung tissues of smokers,and alveolar wall became capsular to expand.Respiratory bronchioles were cystic expansion and small bronchial wall becomed thickening,with severe bronchial epithelial membrane ranking nuts-chaos and peeling.Goblet cells and the cell surface of the cavity mucous secretions and suppositories were more than those of non-smokers.The P53 protein was expressed in cell nucleus and VEGF expressed in the cytoplasm and endothelial cells of neovascularization.Both their positive behavior was granular brown.Either for smoking or for non-smoking lung cancer,the expressions of P53 and VEGF were higher in tumor tissues than that of in the pericancer and surrounding lung tissue(P

Objective To explore the best way for clinical screening of primary aldosteronism (PA).Methods Three hundred and three suspected cases of PA were collected and divided into groups of primary aldosteronism group, essential hypertension group, and nonsecreting cortical adrenal tumor group.The plasma aldosterone concentration/plasma renin concentration ratio ( ARR) was used to draw the receiver operating characteristic ( ROC) curve and obtain the best cut-off point.Furthermore, the current screening schemes for PA were compared.Results Upright ARR yield had larger areas under the ROC curve than plasma aldosterone concentration or plasma renin concentration under all conditions of testing. The best cut-off point of upright ARR[(pg/ml)/(μIU/ml)] for the diagnosis of PA was 43.45.During the two postural stimulation tests,the two upright ARR exceeded 43.45 with the highest diagnostic sensitivity of PA reaching 0.94.During the two upright tests ARR was less than 43.45, with a sensitivity of 0.74, and a specificity of 0.94.Conclusion To screen for PA in high-risk populations, twice postural stimulation test is recommended.As long as the upright ARR is above 43.45, PA may be considered and further confirmation is needed to prevent misdiagnosis.

Objective To investigate the roles of protein kinase C(PKC)β_2 and PPARα in the mRNA expression of vascular endothelial growth factor(VEGF)and vascular cell adhesion molecule-1(VCAM-1)in human umbilical vein endothelial cells(HUVECs)exposed to high glucose,and to explore their relationship.Methods The HUVECs were divided into eight groups:normal glucose(NG,5 mmoL/L D-glucose)group,high glucose(HG,25 mmol/L D-ucose)group,osmotic control(L,NG+20 mmol/L L-glucose)group,normal glucose transfected with empty vector(NN,NG+AdS-null)group,high glucose transfected with PKCβ_2(HB,HG+Ad5-PKCβ_2)group,high glucose plus fenofibrate(HF,HG+40 μmol/L fenofibrate)group,and HB plus fenofibrate (HBF,HB+40μmol/L fenofibrate)group.HUVECs were incubated with fenofibrate for 20 minutes as HF20 group.All cells in various groups were cultured for 6 days.The expressions of VEGF and VCAM-1 mRNA were detected by RT-PCR.PPARα protein expression was tested by Western blot.The expression and traaslocation of PKCβ_2 protein were observed by confocal laser scanning microscopo.Results (1)HG increased VEGF and VCAM-1 mRNA expressions,with 1.91 and 1.56 folds of NG group,respectively(both P<0.05).VEGF and VCAM-1 mRNA expressions in HB group further increased,with 2.59 and 2.07 folds of NG group,respectively(both P<0.05).Fenofibrate significantly decreased VEGF and VCAM-1 mRNA expressions,with 68%and 74%of HG group,respectively(both P<0.05).There were no significant difierences in the expressiong of VEGF,VCAM-1 mRNA between HF20 and HG groups.(2)The protein expression of PPARα decreased by 20%in HG group compared with NG group,and further decreased in HB group,being 78%of HG group.Compared with HG group,PPARαexpression increased by 13%in HF group(P<0.05).(3)HG induced PKCβ_2 translocation from cytosol to nucleus and quantitative analysis showed the ratio of plasma/nuclear fluorescence intensity in HG group decreased by 37% compared with NG group(P<0.05).The PKCβ_2 translocation was more obvious in HB group than in HG group.Conclusion Hiigll glucose stimulates VEGF and VCAM-1 mRNA expressions in HUVECs via PKCβ_2 activationdependent PPARα pathway.

Objective To test the RET-proto-oncogene in a multiple endocrine neoplasia type 2( MEN2) family for confirming the diagnosis and classification , guiding treatment and prevention , and improving the prog-nosis.Methods There were 2 patients of MEN2 with clinical diagnosis and 1 asymptomatic first-degree relative in the pedigree .PCR and direct gene sequencing of PCR produces were used to scan the entire 21 exons of RET-proto-oncogene in the 3 members of the pedigree and 3 normal controls .Results The 2 patients and 1 asympto-matic first-degree relative in the pedigree all had a mutation of the codon 634 in exon 11.It was a heterozygous missense mutation C634R(TGC → CGC).The 3 normal controls showed no abnormalities .Conclusion The gene test of RET proto-oncogene helps to confirm the diagnosis of the pedigree as MEN 2, which can guide the treatment and help identify one asymptomatic mutation carrier .

This paper analyzes the recently epidemic status of schistosomiasis,the change of natural and social factors,and field survey and evaluation data of schistosomiasis in Ya’an City after Lushan Earthquake on April 20,2013,and proposes that it is necessary to strengthen the conventional schistosomiasis control measures,the control of exogenous infection sources,the con-trol of Oncomelania hupensis snails and health education for ensuring no major epidemics after the disaster. This paper also recom-mends the direction and suggestions for future schistosomiasis control in Ya’an City.

Objective To evaluate the effects of measures on the schistosomiasis control after the earth quake in Lushan County so as to provide the experiences for post-disaster schistosomiasis control. Methods The measures taken in schistosomi-asis control after the earth quake were reviewed in Lushan County in 2013 and the epidemic situation of schistosomiasis was in-vestigated and the results were analyzed. Results The schistosomiasis control in floating population and the control of Oncome-lania hupensis snails were enhanced and no schistosome infections were found in both human and livestock. No infected snails and infested water were found. Conclusion The measures of schistosomiasis control after the disaster are effective in Lushan County and the goal to prevent major plague after the earth quake is achieved.

Objective To investigate the prevalence of diabetes (DM) in a community of Chongqing downtown area in 2008.Methods The investigation was conducted from March to June 2008.Cluster sampling in a population aged above 20 years was applied in a community of Chongqing downtown area.3668 subjects were enrolled by oral glucose tolerance test (OGTF).Results The prevalence rate of DM,IGT and IFG was respective 7.44%,6.43% and 7.16%.All the prevalence rates were inereased with aging.Conclusions The incidence of DM in Chongqing city is relatively high,and the incidence is increasing with time.The prevention may be strengthened.