OBJECTIVE:To provide a reference for promoting the effect of government subsidies to the R&D conducted by pharmaceutical enterprises,increasing enterprises’internal input to R&D and improving innovation performance. METHODS:By building the theoretical model of the relation of the government subsidies and internal input to R&D and the innovation performance on the basis of the assumptions,the empirical study was conducted on the pharmaceutical enterprises in Jiangsu Province though re-gression analysis and mediating effect analysis. RESULTS & CONCLUSIONS:The scale designed had good reliability and validity. The final model was obtained after empirical test. Giving considerations to relevant coefficients,the results showed that all assump-tions were verified except 3. Where,government subsidies to the R&D promoted enterprises’internal input to R&D and innovation performance,and internal input to R&D served as a complete mediator between government subsidies to the R&D and innovation performance,that is to say,government subsidies to the R&D had indirect promotion influence on innovation performance,with the influence coefficient as 0.405. It is suggested that the government should increase subsidies to R&D conducted by pharmaceuti-cal enterprises in a reasonable manner to make full use of the guiding function of government funds,and pharmaceutical enterprises should attach importance to internal input to R&D and establish its principal status in innovation system.

Objective:To compare the human ??T lymphocytes expanded with HSP 70BCG and HSP 70 m-peptide complexes with those expanded with solid phase antibody in vitro.Methods:PBMCs were cultured with anti-TCR ?? antibody?HSP 70BCG and HSP 70 m -peptide complexes.The total cell number was counted.The flow cytometer was used to analyze the lymphocytes phenotypes and subtypes.RT-PCR was used to analyze the level of V? mRNA.The cytotoxitic activity of ??T cells against Daudi was determined using MTT calorimetric assay.Results:After 14 days of culture,in the antibody culture system,the total cell number increased about 30-40 fold,and the ratio of ??T cell reached to 86.3%;In the HSP 70BCG culture system,the total cell number increased about 7-8 fold,and the ratio of ??T cell reached to 71.23%;In the HSP 70-peptide complexes culture system,the total cell number increased about 4-5 fold,and the ratio of ??T cells reached to 27.26%.The antibody and HSP 70BCG activated ??T cells possessed a whole repertoire and mainly expressed V?9/V?2 subset and exhibited a strong cytotoxicity against Daudi.Conclusion:Anti-TCR ?? antibody and HSP 70BCG were able to proliferate purer ??T cells.The cells had a whole repertoire,and exhibited strong cytotoxicity against Daudi.

New epidemic broke out in recent year which was suspected to be caused by variant Muscovy duck parvovirus (MDPV). For this reason, new MDPV detection methods are needed for the new virus strains. In this study, a pair of primers were designed according to the full-length genome of MDPV strain SAAS-SHNH, which were identified in 2012, and were used to amplify the vp3 gene of MDPV by polymerase chain reaction. After being sequenced, the vp3 gene was subcloned into the prokaryotic expression vector PET28a. The recombinant plasmid was transformed into E. coli BL21 and induced with IPTG. SDS-PAGE and Western blotting analysis showed the MDPV vp3 gene was successfully expressed. After being purified by Ni2+ affinity chromatography system, the recombinant protein was used as antigen to immunize rabbits to obtain antiserum. Western blotting analysis showed that the acquired antiserum could react specifically with VP3 protein of J3D6 strain and MDPV vaccine strain. The antiserum could also be used for detection of cultured MDPV from primary duck embryo fibroblasts by immune fluorescence assay (IFA). It could be concluded that the VP3 protein and its antibody prepared in the research could be used for detection of VP3 antiserum and antigen respectively.
Animals ; Blotting, Western ; DNA Primers ; Ducks ; virology ; Electrophoresis, Polyacrylamide Gel ; Immune Sera ; biosynthesis ; Parvovirus ; Polymerase Chain Reaction ; Rabbits ; Recombinant Proteins ; genetics ; Viral Proteins ; genetics

Aliphatic amines in infant food packaging materials were extracted and concentrated by 0 . 5 mL of acidified methanol using gas purge microsyringe extraction ( GP-MSE ) . Pre-column fluorescence labeling of amines was achieved in mild conditions with 10-ethyl-acridine-2-sulfonyl chloride ( EASC ) as labeling reagent. The derivatization was carried out at 60℃ and pH 10. The derivatives were successfully separated on a Hypersil GOLD column with excitation and emission wavelengths of 262 and 430 nm, respectively. The detection limits were in the range of 0. 4-0. 6 μg/kg, and the quantitation limits were in the range of 1. 2-2. 1 μg/kg. All analytes were in good linearity in the concentration range of 2. 0-2000 μg/L with correlation coefficients of higher than 0. 998. The developed method was characterized by celerity, accuracy and high sensitivity. It was successfully applied to the determination of aliphatic amines in infant food packaging materials.

ObjectiveTo construct the eukaryotic expression plasmids containing cysteine protease inhibitor (CPI) and glyceraldehydes-3-phosphate dehydrogenase (GAPDH) gene from periodic Brugia malayi (Bm),and to observe its cellular immune response in mouse.Methods pcDNA3.1 (+)-BmCPI/BmGAPDH was constructed.The recombinant plasmids were screened and identified by digestion with restriction enzyme.BALB/c mice were injected intramuscularly with a dosage of 100 μg purified recombinant plasmid DNA with GpG oligodeoxynucleotide (CpG ODN) and two same doses were administrated at 2-week intervals.pcDNA3.1 (+) and phosphate buffered solution (PBS) were used as controls.The tissue of muscles at 4 weeks after the third injection was collected and the target gene was detected by reverse transcription-polymerase chain reaction (RTPCR).Two weeks after the third immunization,the stimulation index (SI) of spleen lymphocytes of immunized mice was measured by methylthiazolyldiphenyl-tetrazolium bromide (MTT) method and the serum levels of interleukin (IL)-4 and interferon (IFN)-γ were detected by enzyme-linked immunosorbent assay (ELISA).The data were analyzed by t test.ResultsBmCPI/BmGAPDH gene in the injected muscle of the immunized mice was detected by RT-PCR. At 6 weeks after immunization,the SIot spleen T lymphocytes in pcDNA3.1 (+)-BmCPI/CpG group and pcDNA3.1 (+)-BmCPI/BmGAPDH/CpG group were 1.466 ± 0.635 and 1.610 ± 0.112,respectively,which were both higher than PBS group and pcDNA3.1( +)-CpG group (1.004 ± 0.019 and 1.078 ± 0.129,respectively) (t=64.438,45.318,42.749 and 34.314,respectively; all P＜0.05).At 4 weeks after immunization,the serum levels of IL-4 and IFN-γ of mice in pcDNA3.1 ( + )-BmCPI/BmGAPDH/ CpG group were significantly higher than those in pcDNA3.1 (+)-CpG group (t=288.053 and 76.453,respectively; both P＜0.05),while the serum level of IFN-γ was also higher than that in pcDNA3.1 (+)-BmCPI/CpG group (t=129.642,P＜0.05). ConclusionThe recombinant eukaryotic plasmid pcDNA3.1 (+)-BmCPI/BmGAPDH could be expressed in mice,and could elicit specific cellular immune responses in immunized mice.

Objective To explore the prognostic value of APACHE Ⅱscore in patients with Severe acute organophosphorus poi-soning .Methods 42 patients with Severe acute organophosphorus poisoning ,in which 34 cases survived ,8 cases dead ,were select-ed .The APACHE Ⅱscores of patients in first 24 h of admission were collected ,and receiver operating characteristic curves (ROC curve) were drawn .Results APACHE Ⅱ score of the 42 patients with Severe acute organophosphorus poisoning was 18 ~30 (20 .11 ± 6 .32) ,in which the survival group was(16 .10 ± 3 .12) ,the dead group was(28 .01 ± 4 .46) (P<0 .01) .With the increase of APACHE Ⅱ score ,the fatality rate gradually increased .The total area under the ROC curves of APACHE Ⅱ score for death judgment was 0 .922 ,APACHE Ⅱ score of 21 .2 was the best diagnostic point ,the sensitivity was 95% ,and specificity was 89% . Conclusion The APACHE Ⅱscore could predict severity of patients with Severe acute organophosphorus poisoning ,and APACHEⅡscore ≥21 .2 could be used as the prognosis for death of the patients .

Objective To observe the therapeutic effect of heat-clearing and dampness-removing therapy combined with Bifico for ulcerative colitis ( UC) induced by trinitrobenzene sulfonic acid ( TNBS), and to explore its influence on tumor necrosis factor alpha ( TNF-α) and interleukin-10 ( IL-10) in rats. Methods Forty female adult Sprague-Dawley rats were evenly randomized into five groups, namely normal control group, model group, Bifico (175 mg/kg) group, Chinese medicine group(enema with Changdiqing 3.6/kg and oral use of Changyanling Recipe l) , and Chinese medicine plus Bifico group. After treatment, the damage of colonic mucous membrane was evaluated, and expression levels of IL-10 and TNF-α in colonic mucosa were observed through immunohistochemical assay. Results The degree of colonic mucosal injury was severer and the inflammation was more obvious in the model group than those in the normal control groups ( P<0.01) , and the above changes were relieved to various degrees in the medication group ( P<0.01 compared with those in the model group) . Chinese medicine plus Bifico group had better effect on reducing colonic mucosa damage index ( CMDI) and inflammation, and on promoting the healing of colonic mucosa than Bifico group and Chinese medicine group (P<0.05 ) . The expression level of TNF-α in the colonic mucosa was markedly increased while that of IL-10 was markedly decreased in the model group ( P<0.01 compared with those in the normal control group) . The medication groups could counteract the above changes in the colonic mucosa ( P<0.01 compared with those in the model group) . The combination group and Chinese medicine group had better effect on decreasing TNF-α expression level and on increasing IL-10 expression level than Bifico group ( P<0.05) . Conclusion IL-10 and TNF-α play an important role in the pathogenesis and development of ulcerative colitis. Chinese medicine combined with Bifico has satisfactory therapeutic effect on UC rats, and its mechanism may be related with the increase of IL-10 expression level and with the decrease of TNF-α expression level.

Objective To systematically review the effectiveness and safety of pantoprazole ( PAN ) vs. ranitidine (RAN) for patients with gastroesophageal reflux disease (GERD). Methods PubMed,Medline,EMbase,The Cochrane Library and three Chinese literature databases (CNKI,VIP and Wan fang) were retrieveed.Randomized controlled trials (RCTs) which compared the clinical outcomes of PAN group vs. RAN group for GERD were included. Two reviewers independently screened literatures in accordance with the inclusion and exclusion criteria, extracted the data and assessed the methodological quality of included studies.Then,meta-analysis was performed using RevMan 5.2 software. Results A total of 8 RCTs involving 1 590 patients were included.The results of meta-analysis showed that the PAN group was significantly superior to RAN group in terms of the healing rates and the relief rates of chief symptom for GERD of gradeⅠ-Ⅲ. While there was no significant difference in the incidence of adverse events between the two groups [GradeⅠ,RR=1.17,95%CI (0.80,1.70),P=0.43;GradeⅡorⅢ, RR=0.76,95%CI (0.43,1.36);P=0.36]. Conclusion Current evidence indicates that,pantoprazole is more effective than ranitidine for GERD of grade Ⅰ-Ⅲ,but both treatments are safe and well tolerated.

Objective:To know the work status of clinical pharmacy in medical institutions of Guizhou province. Methods:Ques-tionnaires were used to analyze the situation of clinical pharmacy in 108 medical institutions of Guizhou province. Results: A total of 246 questionnaires were taken back, and among the 231 valid questionnaires were received including gradeⅡor above hospitals. The main contents of clinical pharmacy work carried out in medical institutions included 7 aspects: pharmacists ’ participation in ward rounds, which accounted for 47. 11%; pharmacists’ participation in case consultation, which accounted for 16. 65%; pharmacists’ participation in teaching practice, which accounted for 38. 84%; pharmacists’ participation in prescription evaluation and analysis, which accounted for 72. 73%;pharmacists’ participation in antimicrobial drug monitoring and drug use evaluation, which accounted for 62. 37%;pharmacists’ participation in drug counsultation and education, which accounted for 58. 68%;pharmacists’ participation in adverse drug reaction monitoring and supervision, which accounted for 77. 32%. Conclusion:The development of clinical pharmacy in Guizhou province still lags behind, and the number of clinical pharmacists is insufficient, which can’ t meet the growing demand for personalized medicine. In particular, the development of clinical pharmacy is restricted by the limited pharmaceutical service. The cog-nition degree of pharmacist group in Guizhou province has been improved. However, the number and the service quality of clinical pharmacists need to be improved further.

Polyethylenimine (PEI) is one of the most characterized non-viral vectors. It can condense DNA in a good manner and achieve high transfection efficiency. Minicircle DNA (mc-DNA) is a novel kind of supercoiled DNA which is devoid of bacterial backbone. mc-DNA is superior to conventional DNA for its higher transfection efficiency and longer time-span. In this study, we combined PEI and mc-DNA in gene delivery system. We investigated the physicochemical and biochemical effects of this non-viral system and further explore its potential in tumor gene therapy. mc-DNA was obtained by recombination of parental plasmid in the presence of L-arabinose, and complexed with PEI. The results of transmission electron microscopy and scanning electron microscopy showed that the particles were spherical and homogeneous. Through gel retardation assay and MTT assay, we found that there were no obvious differences in binding capability of PEI to mc-DNA and plasmid DNA, as well as in cytotoxicity. The results of dynamic light scattering showed that the size of PEI/mc-DNA was about 68 nm, a slight larger than that of PEI/plasmid DNA. Furthermore, the tumor cells transfected with mc-GFP showed higher GFP expression level than that of conventional plasmid. The same results were achieved in the cells treated with tumor-suppressor gene pten, assayed by RT-PCR and Western blot. It indicates that the system of PEI/minicircle DNA is promising in gene transfer.
DNA, Circular ; genetics ; Gene Transfer Techniques ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; Nanoparticles ; chemistry ; PTEN Phosphohydrolase ; genetics ; Polyethyleneimine ; metabolism ; Transfection