Objective To investigate the predictive value of symmetrical ambulatory arterial stiffness index (S-AASI) in detecting early renal impairment of patients with essential hypertension.Methods Totally 245 consecutive out-patients were confirmed with essential hypertension,and were divided into group A (56 cases),group B (64 cases),group C (72 cases),and group D (53 cases) according to the quartile of S-AASI.The combination testing of serum cystatin C,serum β2-microglobulin as well as urine microalbumin to creatinine ratio were implemented as laboratory diagnosis index of renal impairment in early stage and 109 essential hypertension patients were diagnosed with early renal impairment.The parameters were compared among 4 groups.Pearson correlation analysis and partial correlation analysis were performed to confirm the relationship between the markers of early renal impairment and S-AASI.The predictive value of S-AASI to detect early renal injury was evaluated by analyzing Receiver Operating Characteristic (ROC) curve.Results With the rising of S-AASI,age as well as 24 hours mean systolic blood pressure (24 h SBP),serum cystatin C,serum β2-microglobulin,urine microalbumin to creatinine ratio and the incidence rate of early renal injury went notably higher while estimated glomerular filtration rate (eGFR) decreased significantly.After controlling for age,correlation test showed S-AASI was positively correlated with24hSBP,serum cystatin C,serumβ2-microglobulin,urine microalbumin to creatinine ratio(r =0.392,0.627,0.514 and 0.643 respectively,P ＜ 0.05) and negatively correlated with eGFR(r =-0.312,P ＜ 0.05).The 24 hours mean diastolic blood pressure (24 h DBP) was uncorrelated with S-AASI.Area under ROC curve of S-AASI for diagnosis of hypertensive renal impairment was 0.885.The critical value of S-AA-SI was 0.17,the sensitivity,specificity,positive predictive value,and negative predictive value were 92.7％,65.2％,68.5％,and 91.7％,respectively.Conclusions When S-AASI was detected above 0.17,patients with hypertension had a higher risk of renal impairment.Higher S-AASI was correlated with worse early renal impairment laboratory indexes.The predictive accuracy of S-AASI for early hypertensive renal impairment was medium.

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To express Pleurocidin in Escherichia coli and to enhance the secretory efficiency of the fusion protein, the gene encoding Pleurocidin was ligated with Cherry DNA sequence via blunt-end ligation. Then this fusion gene was cloned into pET22b (+) vector and the recombinant plasmid was transformed into E. coli BL21 (DE3). Lactose was used to induce expression of fusion protein. The recombinant plasmid pET22b (+) -CP was successfully constructed and high-level expression of fusion protein was induced with lactose. Statistics showed that addition of glycine after 16 h of induction significantly enhanced the secretory efficiency of the fusion protein. After hydrolysis of the fusion protein by diluted hydrochloric acid and some further purification steps, r-Pleurocidin was obtained with antibacterial activity against E. coli DH5α and Bacillus subtilis BS168. In conclusion, the fusion protein was expressed in E. coli and biologically active r-Pleurocidin was obtained after hydrochloric acid cleavage and purification.
Animals ; Cloning, Molecular ; Escherichia coli ; metabolism ; Fish Proteins ; biosynthesis ; Flounder ; Recombinant Fusion Proteins ; biosynthesis

Objective:To investigate effects of a novel synthetic immunostimulator CH1b containing thiazolidin-4-one on the immunoregulation funotion of iNKT ( invariant nature killer T ) cells in active RA patients in vitro.Methods: Peripheral blood mononuclear cells( PBMCs) isolated from active RA patients were cultured with stimulation of α-Galcer and IL-2 in vitro and iNKT cells were then separated by using magnetic activated cell sorting( MACS) method with iNKT isolation kit.The cells were divided into three groups:control group (IL-2),α-Galcer group (IL-2+α-Galcer),CH1b group(IL-2 +CH1b).The effects of CH1b on the proliferation of iNKT cells in active RA patients were analyzed by using MTT assay.MILLIPLEX MAP Human Cytokine/Chemokine kit was used to evaluate the secretion of IFN-γand IL-4 in iNKT cells culture media.The expressions of IFN-γmRNA and IL-4 mRNA in iNKT cells were analyzed by RT-PCR.Results: Compared with control and α-Galcer group,the proliferation of iNKT cells of CH1b group were significantly higher( P<0.05).Compared with control,the ratio of IFN-γ/IL-4 in iNKT cells culture media in active RA patients of CH1b group were significantly lower (P<0.05).Compared with control,expressions of IFN-γmRNA and IL-4 mRNA were higher inα-Galcer group;compared with control,expressions of IL-4 mRNA were higher in CH1b group,while there were no obvious difference on expressions of IFN-γmRNA.Conclusion:CH1b was found to significantly stimulate the actived iNKT cells in active RA patients proliferation,promote the secretion of IL-4,and increase the ratio of IFN-γ/IL-4,promote the expression of IL-4 mRNA in iNKT cells in active patients.

Objective To explore the effects of the Chinese medicine, modified Erchen decoction, on the serum lipid spectrum of ApoE-/-mice, and to explore its possible anti-atherosclerotic mechanism.Methods Forty-four male 7-8-week old ApoE-/-mice were used in this experiment.ApoE-/-mouse models of atherosclerosis were generated by high-cholesterol diet for 4 weeks.And then, they were given simvastatin or modified Erchen decoction by gavage.The body weight of mice was recorded every week, The mice were sacrificed after treated with the drugs for 8 weeks continuously, and the plasma lipid was determined by enzymatic method.The aortic valves and arches were stained with oil red O to depict atherosclerotic plaques and liver structural changes of the mice were examined by pathology.Results Modified Erchen decoction lowered plasma lipid ( including TCHOL and LDL-C ) significantly ( P<0.01 ) .The body weight was increased in the mice of all groups, but it was more pronounced in the mice of model group than in the blank and modified Erchen decoction groups.The serum CHOL and LDL-C levels were significantly lowered in the modified Erchen decoction group (P<0.01).The area of atherosclerotic plaques in the aortic wall was significantly reduced in the mice of modified Erchen decoction group as shown by oil red O staining.The pathological changes of hepatocytes were less severe and the structure of hepatic lobules was better preserved in the mice of modified Erchen decoction group.Conclusions The Chinese medicin modified Erchen decoction can effectively reduce serum lipids, regulate lipid metabolism, and ameliorate the process of atherosclerosis in ApoE-/-mice.

Objective To establish and evaluate intestinal epithelial barrier model using Caco-2 cell so as to play a foundation for next study of barrier permeability.Methods Caco-2 cells were cultured in vitro then seeded into Transwell cell culture inserts.The permeability of the intestinal epithelial barrier was detected by transepithelial electrical resistance(TEER)and lucifer yellow flux,and verified by transmission electron micro-scope.Different concentrations of PAF(0,50,100,and 200 nmol /L)were exposed for 24 hours to Caco-2 mono-layer when cultured 21 days.The tight junction was observed under transmission electron microscope.Assess-ment of ZO-1 protein localization and expression were detected by immunofluorescence and Western blot analy-sis.Results Cultured Caco-2 cell confluencd as monolayer with time passed.From 5th day,TEER increased, then reached 600Ω?cm2 at 15th day and lasted to 21 st day,there was little flux of lucifer yellow,transmission e-lectron microscopy also found cells differentiated better,had well-arranged villi and polarity alined as monolayer, forming completed tight junction which was the marker of intestinal epithelial barrier model in vitro.TEER de-creased and lucifer yellow flux increased in cells exposed to PAF.The permeability reached the peak when ex-posed to 100 nmol /L PAF(P <0.01 ),tight junction disrupted,ZO-1 protein expression downregulated,abnor-mal localization and distribution was assessed by immunofluorescence staining.Conclusion Cultured Caco-2 cells for 2-3w can be used to study intestinal epithelial barrier as a model in vitro.PAF increased intestinal epi-thelial permeability,which would correlate to the decreased protein expression and abnormal distribution of ZO-1.

This study investigated the anti-HSV-2 effect of alumen through in vitro and in vivo experiments. Viable cell counting was employed to assess the toxicity of alumen on Vero cells. The inhibition rate of HSV-2 was defined as the cytopathic effect (CPE) of the cells infected with the virus. Alumen suppositories of different concentrations were vaginally applied to the guinea pigs which were then infected with HSV-2 via a vaginal route. The clinical symptoms were observed and the local virus titer calculated. The results showed that alumen had an in vitro anti-HSV-2 effect by means of antiviral duplication, direct killing of the virus, and antiviral adsorption. Alumen suppositories of different concentrations could reduce or completely inhibit HSV-2 infection in guinea pigs. It was concluded that alumen had an in vitro anti-HSV-2 effect through multiple approaches and it could suppress in vivo vaginal HSV-2 infection of guinea pig to some extent.

Objective To determine the efficacy of primary tumor of esophageal cancer,according to the result of magnetic resonance imaging before and after chemoradiotherapy of esophageal cancer,combined with clinical efficacy evaluation,and to verify the reliable evaluation of the short-term curative effect of magnetic resonance on esophageal cancer,combined with the original CT and esophagogram evaluation criteria.Methods From May 2010 to March 2014,totally 83 patients with esophageal carcinoma treated with 3D-CRT or IMRT were enrolled.The prescribed doses were ranged from 50-64 Gy with median dose of 60 Gy and 1.8-2.0 Gy per fraction,of which 34 of the patients received concurrent chemotherapy of FP or TP.All the patients performed the examinations of DWI,CT scan and esophagogram before and after radiotherapy.The treatment efficacy was evaluated by short-term therapeutic effect evaluation criterion of versions 1989 and 2013 and the hyperintense expression on DWI sequence.Results According to the short-term therapeutic effect evaluation criterion of versions 1989 and 2013 based on the examination of esophagogram and CT scan,45 patients achieved complete remission (CR) after treatment(54.2％) and 38 achieved partly remission(PR) (45.8％) version 1989,while 35 patients achieved CR (42.2％) and 48 achieved PR (57.8％) version vesion 2013.In the two differentcriterions,the local control rate and survival rate of the complete remission group in 1 to 5 years were better than those in the partial remission group.According to the examination of DWI,48 patients' hyperintense disappeared completely at the end of treatment (which was defined to CR),25 patients had a slightly hyperintense expression and 10 patients still had hyperintense expression on DWI sequence (which two defined to PR),the local control and survival rates of the former group were superior to the latter groups (x2 =6.125,11.652,P ＜0.05).The TE results evaluated by DWI and TE evaluation criterion of version 2013 were compared according to Kappa test,as a result,the Kappa coefficient 0.478.According to the examination of esophagogram,CT scan and DW1,25 patients achieved CR and 58 achieved PR in all exams,and the local control and survival rates of the former group were superior to the latter group (x2 =5.559,10.014,P ＜0.05).Conclusions The esophagogram and CT based TE evaluation criterion could well indicate local control status of esophageal cancer,and the examination of DWI could afford visualized and quantifying reference information about the TE of esophageal cancer.The expression of hyperintense at the end of treatment may indicate a high risk of recurrence and metastasis.The therapeutic effect evaluated by esophagogram,CT scan and DWI maybe more objective and more accurate.

This study investigated the anti-HSV-2 effect of alumen through in vitro and in vivo experiments.Viable cell counting was employed to assess the toxicity of alumen on Vero cells.The inhibition rate of HSV-2 was defined as the cytopathic effect (CPE) of the cells infected with the virus.Alumen suppositories of different concentrations were vaginally applied to the guinea pigs which were then infected with HSV-2 via a vaginal route.The clinical symptoms were observed and the local virus titer calculated.The results showed that alumen had an in vitro anti-HSV-2 effect by means of antiviral duplication,direct killing of the virus,and antiviral adsorption.Alumen suppositories of different concentrations could reduce or completely inhibit HSV-2 infection in guinea pigs.It was concluded that alumen had an in vitro anti-HSV-2 effect through multiple approaches and it could suppress in vivo vaginal HSV-2 infection of guinea pig to some extent.

Objective To examine the prediction value of diffusion-weighted magnetic resonance imaging (DWI) on radiotherapy response in esophageal cancer.Methods A total of 24 subcutaneous esophageal cancer xenograft models were randomly divided into experimental group (n =14,received a single dose of 15 Gy radiotherapy) and control group (n =10,without any treatment).MRI were required before and after radiotherapy at different check time points (1,6,13 days) of T1WI,T2WI,and DWI measurements.Apparent diffusion coefficient (ADCX) and volume (VX) of each xenograft were measured,and both △ADCX and △VX were calculated.Results The ADC values of both group were decreased at the first day,however,the decrease in experimental group were more obviously with an increase at 6 and 13 d gradually.However,the ADC values of the control group showed a persistent decline.There was no significant difference in the ADC values between the two different groups before radiotherapy (P ＞ 0.05),while significant difference was found in the ADC values (F =6.178,16.181,58.733,P ＜ 0.05) and △ADC after radiotherapy (F =9.038,12.360,35.140,P ＜ 0.05).The xenografts volume in the experimental group showed a significant growth delay.There was no significant difference in volume between the two groups (P ＞ 0.05) before radiotherapy.Significant difference in V between the two groups only began to exist at 5 d after radiotherapy (F =28.587,P ＜ 0.05).The ADC0,ADC1 of transplanted tumor in control group had linear correlation relationships with its volume of later period.After radiotherapy,the trend of r values gradually increased from-0.118 to 0.896.Conclusions ADC values may change significantly at the early stage after radiotherapy,and initial and early ADC value may have close relationship with xenograft volumes of later period,which indicates that DWI has huge potential in the prediction of radiotherapy response.