[ ABSTRACT] AIM:To investigate the effect of Jia-jian-yi-yin decoction on endothelium-dysfunction in ovariecto-mized rats.METHODS:The ovariectomized rats were treated with Jia-jian-yi-yin decoction or turbid liquid of estradiol va-lerate for 8 weeks.The vascular ring tension was measured.Scanning electron microscopy and Western blotting were ap-plied to assess the function of endothelium-dependent dilation, aortic endothelial morphology and the expression of endothe-lial lipase.The pathologic sections were prepared to observe the effect of Jia-jian-yi-yin decoction on the uterus.RE-SULTS:In ovariectomized rats, the decrease in endothelium-dependent relaxation to acetylcholine ( ACh) was reversed to normal level, the endothelial morphology returned to normal without lipid accumulation and the endothelial lipase expression was decreased by Jia-jian-yi-yin decoction.Furthermore, no obvious change of the wet weight of uterine between the ovari-ectomized rats with or without Jia-jian-yi-yin decoction treatment was observed.CONCLUSION:Jia-jian-yi-yin decoction may have protective effects on endothelium-dependent vasodilation and aortic endothelial morphology in estrogen-deficient animals.

Although DNA vaccination is now a promising strategy against hepatitis B virus (HBV) infection, this approach has relatively modest antiviral effect, indicating that immunosuppressive mechanisms may occur in the long-term established infection. In this study, we studied the immunogenicity and anti-HBV efficiency of a combination of HBV surface (HBsAg) and core (HBcAg) DNA vaccine, enhanced by heat shock protein (HSP) gp96 or HSP70 and mediated by in vivo electroporation. Immunization with gp96 adjuvanted HBsAg/HBcAg DNA formulation induced potent T cell and antibody immunity against HBsAg and HBcAg. Notably, treatment with gp96 or HSP70 as adjuvant resulted in reduction of Treg populations by around 20%. Moreover, compared with nonimmunized control mice, immunization with gp96 or HSP70 adjuvanted DNA vaccine dramatically decreased serum HBsAg and viral DNA levels, and HBcAg expression in liver. These results may therefore provide an effective strategy for designing gp96-based DNA vaccine for immunotherapy of chronic HBV infection.
Adjuvants, Immunologic ; Animals ; Electroporation ; HSP70 Heat-Shock Proteins ; immunology ; Hepatitis B Core Antigens ; immunology ; Hepatitis B Surface Antigens ; immunology ; Hepatitis B Vaccines ; immunology ; Hepatitis B, Chronic ; prevention & control ; Immunization ; Membrane Glycoproteins ; immunology ; Mice ; Mice, Transgenic ; Vaccines, DNA ; immunology

Objective To investigate the time-effect relationship of angiogenesis induced by ultrasound.mediated microbubble destruction in the skeletal muscle of rats.Methods Forty-eight healthy SD rats were divided into 4 groups:ultrasound-mediated microbubble destruction group,ultrasound only group,microbubble only group and control group.In ultrasound-mediated mierobubble destruction group, microbubbles were inj ected by vein at a dose of 0.5 ml and the target skeletal muscle was radiated at 2.0W/cm2.In ultrasound only group,the target skeletal muscle was radiated at 2.0 W/cm2.In microbubble only group.microbubbles were injected by vein at a dose of 0.5 ml.The control rats were without ultrasound radiation and microbubbles.On the 3rd,7th,10th,14th,21st and 28th day after the ultrasound radiation,two rats in each group were sacrificed and the target skeletal muscle was harvested for HE staining to observe the microstructure of tissue,immunohistochemistry staining was used to count the microvessel density (MVD),enzyme 1inked inmmunosorbent assay(ELISA)was used to detect the expression of vascular endothelial grouth factor(VEGF).Results Angiogenesis was significant in ultrasound-mediated mierobubhle destruction group,but a little in ultrasound only group.There was not any angiogenesis in either microbubble only group or control group.MVD and VEGF expression of ultrasound-mediated microbubble destruction group arrived at a peak on the 1Oth day as well as on the 14th day of ultrasound group.Conclusions Ultrasound-mediated microbubble destruction can facilliate the endogenous secretion of VEGF quickly and more,and accelerate the angiogenesis in the skeletal muscle.

Objective To compare the clinical effect of neoadjuvant chemoradiotherapy (nCRT) and neoadjuvant chemotherapy (nCT) in the treatment of locally advanced esophageal squamous cell carcinoma.Methods The retrospective cohort study was conducted.The clinicopathological data of 156 patients with local advanced esophageal squamous cell carcinoma who were admitted to the Zhongshan Hospital of Fudan University from January 1,2010 to December 31,2015 were collected.Among 156 patients,59 undergoing nCRT were allocated into the nCRT group and 97 undergoing nCT were allocated into the nCT group.Patients in the nCRT group and nCT group respectively received 2 cycles chemotherapy by the TP regimen+40 Gy radiotherapy (2 Gy/d) and 2 cycles chemotherapy by the TP regimen.Patients were evaluated by imaging examinations after 6 weeks neoadjuvant therapy completion,and then underwent abdominal and right chest-left cervico three-incision thoracoscopic surgery.Observation indicators:(1) treatment situations;(2) postoperative pathological examination;(3) follow-up and survival situations.Follow-up using outpatient examination and telephone interview was performed once every 3 months within 2 years and once every 6 months after 3 years up to January 2017.Follow-up included levels of tumor markers [carcinoembryonic antigen (CEA) and SCC-Ag],thoracic or abdominal computed tomography (CT),neck and abdominal ultrasonography and gastroscopy or PET/CT examination if necessary.Measurement data with normal distribution were represented as (x)±s and comparison between groups was analyzed using the t test.Measurement data with skewed distribution were described as M (range) and comparison between groups was analyzed using the nonparametric test.Count data were analyzed using the chi-square test or Fisher exact probability.Comparison of ordinal data was done by the nonparametric test.The survival rate was calculated using the life table method and survival was analyzed by the Log-rank test.Results (1) Treatment situations:all the patients in the 2 groups were able to burden neoadjuvant therapy and thoracic esophagectomy.Six patients in the nCRT group and 15 in the nCT group had conversion to open surgery.Operation time,volume of intraoperative blood loss,cases with postoperative readmission of ICU,cases with complications,cases with perioperative death and duration of hospital stay were (201 ± 25) minutes,(137± 66)mL,5,24 (10 with pulmonary complications,8 with anastomotic leakage,3 with hoarseness,2 with cardiovascular complications and 1 with chylopleura),0,12 days (range,9-93 days) in the nCRT group and (195±20) minutes,(133±58) mL,8,30 (11 with anastomotic leakage,10 with pulmonmy complications,4 with hoarseness,2 with cardiovascular complications,1 with postoperative hemorrhage,1 with delayed gastric emptying and 1 with chylopleura),1,11 days (range,9-78 days) in the nCT group,respectively,with no statistically significant difference between the 2 groups (x2 =0.883,t =0.102,0.692,x2 =0.048,1.541,Z =0.225,P＞ 0.05).(2) Postoperative pathological examination:R0 resection rate was 96.6％ in the nCRT group and 93.8％ in the nCT group,with no statistically significant difference between the 2 groups (x2 =0.589,P＞0.05).Results of postoperative pathological examination showed that G0,G1,G2 and G3 of tumor regression grade were respectively detected in 18,16,7,18 patients in the nCRT group and 4,5,4,84 patients in the nCT group,with a statistically significant difference between the 2 groups (Z=-7.151,P＜0.05).Stage 0,Ⅰ,Ⅱ,ⅢA,Ⅲ B and ⅣA of postoperative ypTNM stage were respectively detected in 16,9,23,4,6,1 patients in the nCRT group and 4,9,37,6,34,7 in the nCT group,with a statistically significant difference between the 2 groups (Z=-4.890,P＜0.05).The down-staging was detected in 48 patients of the nCRT group and 50 patients of the nCT group,with a statistically significant difference between the 2 groups (x2=13.957,P＜0.05).(3) Follow-up and survival situations:of 156 patients,153 were followed up for 12-82 months,with a median time of 36 months.The 1-,3-,5-year overall survival rates were 88.1％,61.4％,34.9％ in the nCRT group and 81.4％,43.8％,23.1％ in the nCT group,with a statistically significant difference between the 2 groups (x2=4.336,P＜0.05).Conclusion The nCRT in the treatment of locally advanced esophageal squamous cell carcinoma can enhance postoperative pathological response rate,down-staging rate and overall survival rate compared with nCT,without increasing incidence of perioperative complications.

Objective To discuss the polymorphisms of asthma susceptibility gene ORMDL3 in infantile wheezing,in order to provide a theoretical basis for early diagnosis of asthma.Methods One hundred and fifty wheezing infants were recruited and divided into 2 groups as asthma predictive index(API) positive group(n =80) and negative group (n =70).Taqman probe was applied to detect the genotypes of 2 single nucleotide polymorphisms (SNPs)in childhood asthma susceptibility gene ORMDL3,which were rs4794820 and rs7216389.The genotype distributions were analyzed and compared between 2 groups,and the correlations among genotype distribution and tidal breath pulmonary function,fractional exhaled nitric oxide (FeNO) concentration,percentage of eosinophils (EOS％),serum immune globulin E (total IgE) levels respectively were also analyzed,respectively.Results (1) The frequencies of rs4794820 AG and rs7216389 TC heterozygotes in the API positive group were the highest,which were significantly higher than those in the negative group(58.75％ vs.31.42％,56.25％ vs.32.86％ respectively,all P ＜0.01).The frequencies of GG and TT homozygotes in the API negative group were the highest,which were significantly higher than those in the positive group (58.57％ vs.30.00％,57.14％ vs.31.25％ respectively,all P ＜0.01).(2)The time to reach the peak expiratory flow in tidal breathing over the total expiratory time (TPTEF/TE) and the volume to reach the peak expiratory flow in tidal breathing over the total expiratory volume (VPEF/VE) of the infants in the API positive group were less than those in the API negative group(16.87 ±5.31 vs.20.12 ± 5.23,20.87 ± 5.92 vs.25.56 ± 6.77,respectively),and the FeNO concentration was higher than that in the API negative group [(22.44 ± 9.77) ppb vs.(13.23 ± 7.90)ppb],and the differences were significant (t =-3.776,-4.490,6.377,respectively;all P ＜ 0.01).(3) In the API positive group,the TPTEF/TE and VPEF/VE of the infants who expressed AG/TC genotype were lower than those who expressed GG/TT genotype (14.55 ± 4.83 vs.19.91 ± 4.17,18.85 ± 4.26 vs.25.20 ± 7.06,respectively,t =-4.727,-3.976,all P ＜ 0.01);while the FeNO concentrations,EOS％ and total IgE levels were higher than those who expressed GG/TT genotype [(25.02 ± 8.77) ppb vs.(18.39 ± 6.56) ppb,7.16 ± 2.62 vs.5.50 ± 1.34,(366 727 ±275 533) IU/L vs.(166 826 ± 62 865) IU/L,respectively] (t =3.484,3.409,4.589 respectively;all P ＜ 0.01).Conclusions Childhood asthma susceptibility gene ORMDL3 SNPs rs4794820 AG and rs7216389 TC heterozygotes are the risk factors for API positive infantile wheezing.The pulmonary function damage and airway inflammation of the infants who expressed AG/TC genotype are more serious than those who expressed GG/TT genotype,and more likely to develop persistent asthma.

Objective To explore the weight for the training target of Applied nursing undergraduate. And find the degree of importance for the indexes of the target , thus highlighting theApplied training objective characteristics. Methods Developed by Bloom's teaching objectives classification theory and educational objectives theory to refined indexes, using Delphi method to establish the Applied nursing undergraduate training objectives. By the importance assignment from experts on each index, using analytic hierarchy process (AHP) method to construct the judgment matrix. Then calculate the weight of each index weight, and test its logical consistency. Results The weight of theattitudes and valueswas 0.56, andknowledgeandabilitywere 0.22. For the secondary indicators, in the values dimension,professional attitudeandideological qualityhad relatively high weight.Under the knowledge dimension, the professional knowledge had the highest weight; and for the ability dimension,communication skillsandapplicationswere heavily maximum. All the random consistency rates (CR) for the indexes were less than 0.10, so there was no logical confusion. ConclusionsAppliednursing undergraduate talents need to combine theory and practice quickly into clinical work. By weight setting, this study provides basis to further quantify and highlight the Applied nursing undergraduate training features, and provides the basis for the application-oriented undergraduate nursing personnel reforms.

Objective To develop the Applied nursing undergraduate students training framework objective, clear the entry of applied training target for undergraduate nursing students. Methods Through literature review and panel discussions, Delphi expert consultation questionnaires were preliminary designed. 23 experts were issued via email and paper questionnaires, conducted two expert advice and analysis. Results A total of 20 valid questionnaires were recovered after two consultation. The expert advice of good scientific reliability:experts had positive reply, and the authority level was 0.857, Kendall were 0.130-0.293, it showed a good level of coordination (P < 0.05). By two rounds of expert advice, three capacity index target level for Applied nursing undergraduate students training were gotten: attitudes, values and basic literacy; knowledge of the target; the ability. The secondary capacity index for target had 15 items, and the third capacity index had 63 items. Conclusions TheAppliednursing undergraduate students training objectives and targets were refined through Delphi which provide the basis for the Applied nursing undergraduate training model and curriculum reform.

Secretory anti-gp96 scFv fragment was expressed in Pichia pastoris to obtain a small molecule antibody that specifically recognizes heat shock protein gp96. The gp96-scFv fragment gene was synthesized and cloned to Pichia pastoris expression plasmid pPICZa-A. Pichia pastoris X33 was electroporated with the linearized recombinant expression vector, and expression of gp96-scFv fragment was induced by methanol. The His-tagged recombinant protein was then purified by affinity chromatography and analyzed with SDS-PAGE and Western blotting assays. The biological activities of recombinant gp96-scFv fragment were determined by Western blotting, Immunofluorescence, ELISA and FACS assays. The gp96-scFv fragment was expressed successfully in Pichia pastoris. About 50 mg of recombinant protein could be purified from 1 liter of the Pichia pastoris culture supernatant. Its molecular weight was about 15 kDa. The gp96-scFv fragment could specifically bind to gp96 protein by Western blotting, immunofluorescence, ELISA and FACS analyses. Pichia pastoris-expressed gp96-scFv fragment specifically recognizes gp96 protein, which could be used for Western blotting, Immunofluorescence, ELISA and FACS analyses.
Blotting, Western ; Chromatography, Affinity ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Membrane Glycoproteins ; immunology ; Pichia ; metabolism ; Plasmids ; Recombinant Proteins ; biosynthesis ; Single-Chain Antibodies ; biosynthesis

Objective To investigate the relationship between high-sensitivity C-reactive protein (hsCRP) and retinal vascular diameter.Methods This study was a cross-sectional study,the population was from the Kailuan study (clinical trial research registration NO.:ChiCTR-TNC-1100 1489) in 2012-2013 (age ≥ 40 years).All the people was checked by cardiovascular and epidemiological examination,of which 3 447 people had an analytical results of fundus color photography.The retinal vascular diameter was analyzed by software.Take the value of hsCRP 1％ to 99％ points and 3 363 people were finally included in the statistical analysis.The mean age of 3 363 people (male 1 880,female 1 483) was 56.69± 10.10 years old.The correlation between hsCRP and central retinal artery equivalent (CRAE),central retinal vein equivalent (CRVE),arteriovenous ratio (AVR) were evaluated by Spearman correlation coefficient.Multivariate linear regression analysis was used to futher analyze the association between hsCRP and retinal fundus diameter parameters.Results The median value of hsCRP was 1.10 (0.50,2.21) mmo/L,CRAE was (174.16±24.36) μm,CRVE was (282.70±26.32) μm,AVR was 0.62-±0.09.Spearman correlation analysis showed that hsCRP was positively correlated with CRVE (r =0.052,P =0.003) and negatively correlated with AVR (r =-0.052,P =0.002).The non-normal variables hsCRP were transformed into continuous variables lghsCRP,and CRAE,CRVE and AVR were used for multivariate linear regression analysis.After a number of factors were gradually corrected,the results showed that association between lghsCRP and CRVE (β =3.428) or AVR (β =-0.006) was statistically significant (P＜0.05).LghsCRP was a risk factor for CRVE both in men and women after the calculation of gender stratification.Conclusions CRVE increased and AVR decreased with the increase of hsCRP.The changes of retinal vascular diameter may be associated with hsCRP as a marker to predict cardiovascular and cerebrovascular disease.