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<p><b>OBJECTIVE</b>To express soluble HA of A/H1N1 influenza virus in drosophila S2 cell line and identify its bio-activity.</p><p><b>METHODS</b>HA gene was amplified from A/Shenzhen/71/09 virus strain using RT-PCR, then we constructed pAC5.1-HA expression vector, which was co-transfected into S2 cell with pCoblast vector. After transfection, stable S2 cell was selected through Blasticindin. HA in the supernatant was identified with Western Blot assay and purified with Ni-column. Recombinant HA was immunized into BALB/c mice 3 times, and the Abs titers were evaluated with ELISA.</p><p><b>RESULTS</b>We successfully cloned HA gene with 1.7 x 10(3) bp of A/Shenzhen/71/09 virus strain and got recombinant pAC5. 1-HA expression vector. Stable S2 cell line was established after transfection and selection, which continuously expressed HA with molecular weight 75 x 10(3) D. After immunization with HA, the Abs titers were 1:1280 and 1: 5120 respectively on 10 d, 30 d.</p><p><b>CONCLUSION</b>We expressed soluble HA with good bio-activity, which contributed to research on immune diagnosis, subunit vaccine, and monoclonal Abs for influenza.</p>
Subject(s)
Animals , Female , Humans , Mice , Blotting, Western , Cell Line , Drosophila , Gene Expression , Hemagglutinin Glycoproteins, Influenza Virus , Chemistry , Genetics , Metabolism , Influenza A Virus, H1N1 Subtype , Genetics , Metabolism , Influenza, Human , Virology , Mice, Inbred BALB C , SolubilityABSTRACT
In this study, the optical data of tongue color of different syndromes in primary hepatic carcinoma (PHC) were detected by optical spectrum colorimetry, and the chromaticity of tongue color was compared and analyzed. The tongue color characteristics of different syndromes in PHC and the relationship between different syndromes and tongue color were also investigated.
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Objective To establish neutralization assay based on H5N1 avian influenza pseudotyped virus in vitro and to evaluate neutralizing titer of convalescent serum from 2 patients with H5N1 avian influenza.Methods pHR-Luc,pCMV△8.2 and CMV/R-SH or CMV/R-TH were cotransfected into 293T cell by co-precipitation with calcium phosphate.Pseudotyped virus supernatant was harvested 72 h posttranofection and identified the expression of HA and P24 by Western blot,and then we analyzed infective activity of 200 μL supernatant of pseudotyped virus.293T cell integrated HA was prepared and anti-HA antibodies in convalescent serum were measured with FACS assay.Neutralizing titers of convalescent serums against Shenzhen and Thailand pseudotyped virus were determined based on calculating IC50 with neutralizing assay.Results Pseudotyped virus involved P24 and HA,and precursor protein HA0 could cleavage into HA1 and HA2 with biological activity.Pseudotyped virus possessed better infective activity,and RLA value was about 2 × 104 with 200 μL supernatant.Both convalescent serums contained anti-HA antibodies and had cross-reactivity against different virus clades with FACS assay.Both convalescent serums had neutralizingactivity and could cross-neutralize different virus clades.However,both serums'neutralizing titers against Shenzhen virus were higher than Thailand.Conclusion We successfully constructed infectious pseudotyped virus which integrated HA of Shenzhen or Thailand virus,and it could be used for evaluation of serum neutralizing activity fast,efficiently and safely with broadly application prospect.
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Objective To observe the impacts of the method of fortifying the spleen and supplementing Qi on the cellular immunity of chronic hepatitis B virus(HBV)carriers.Methods Asymptomatic HBV carriers group(ASC group,n=60)with HBV DNA positive were randomly divided into Fortifying the Spleen and Supplementing Qi decoction group(A group,n=30)and conventional treatment group(B group,n=30),the peripheral blood T lymphocyte subsets were detected with flow cytometey and compared with those of the healthy people(The normal control group,n=18).All the patients were treated,and 24 weeks after treatment the indexes were measured again.Results As compared with those of normal controls,the numbers of CD4+ T cells and CD8+ T cells were decreased significantly in HBV carriers(P＜0.01);There was a statistical significance between the values before and complete the treatment with the decoction of fortifying the spleen and supplementing Qi(P＜0.05 or P＜0.01),and compared with conventional treatment group,there Was a statistical significance(P＜0.05).Conclusion T-cell subset function was disturbed in the HBV carriers.The method of fortifying the spleen and supplementing Qi could improve the cellular immunity of carriers with chronic hepatitis B.
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Objective To compare the clinical efficacy and safety of pegylated interferon α-2a (Peg IFN α-2a) or adefovir dipivoxil(ADV) monotherapy and their combination therapy in HBeAg positive chronic hepatitis B (CHB) patients. Methods An open randomized controlled multicenter clinical trial was performed. One hundred and twenty cases with CHB were divided into 3 groups: Peg IFN α-2a monotherapy (group A), ADV monotherapy (group B) and Peg IFN α-2a plus ADV combination therapy (group C). The virological response (VR), serological response (HBeAg, HBsAg clearance and seroconversion), biochemical response (BR) and sustained response (SR) were tested at week 24 and 48 of therapy and week 48 of follow-up after end of treatment (EOT) for'evaluation of therapeutic effects, safety and drug resistance. The efficacy was compared using X2 test. Results At week 48 of treatment, the VR (HBV DNA ≤500 copy/mL) rates were 36. 8%(14/38), 37. 5%(15/40) and 62. 9% (22/35), respectively in groups A, B and C; that in group C was higher than those in groups A and B (X2 = 4. 933, 4. 801, respectively; both P < 0. 05); HBeAg seroconversion rates in three groups were 44. 7% (17/38), 17. 5% (7/40) and 51. 4% (18/35), respectively. At week 48 of follow-up,SR rates in three groups were 34. 2%(13/38), 15. 0%(6/40) and 48. 6% (17/35), respectively; those in groups C and A were higher than that in group B (X2 = 9. 894,P<0. 01;X2 =3. 903, P<0. 05, respectively). Conclusions VRs at week 24 and 48 of Peg IFN α-2a plus ADV combination therapy are better than Peg IFN α-2a or ADV monotherapy. SRs at week 48 of follow-up after Peg IFN α-2a monotherapy and combination therapy are both better than ADV monotherapy.
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To establish a method based on a pseudotyped virus assay for screening drugs for anti-H5N1 avian influenza virus, and then to evaluate anti-virus activity of traditional Chinese herbal compounds with the function of purgation, detoxification, cooling the blood and reinforcing the healthy qi based on seropharmacology.
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<p><b>OBJECTIVE</b>To introduce the double scanning for producing computer aided design and computer aided manufacture (CAD-CAM) zirconia crowns, and to compare Procera with Everest CAD-CAM system by observing clinical effects of the crowns.</p><p><b>METHODS</b>Forty CAD-CAM zirconia crowns (10 crowns by Procera, and 30 crowns by Everest) were evaluated, and the examination criteria included: marginal fit, aesthetics, gingival health, retention, and fracture. All of the zirconia copings were produced using double scanning. Procera system and Everest system use contact scanner and optical scanner respectively.</p><p><b>RESULTS</b>The observation time for the crowns ranged from 6 to 24 months. The results of clinic evaluation were acceptable in all restorations.</p><p><b>CONCLUSIONS</b>Double scanning could be used to produce complicated zirconia copings by CAD-CAM in order to get proper symmetrical porcelain length. It was easier to make complicated coping with contact scanner than with optical scanner.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Computer-Aided Design , Crowns , Dental Porcelain , Dental Prosthesis Design , Dental Restoration Wear , Follow-Up Studies , Photometry , Surface Properties , ZirconiumABSTRACT
Objective To construct pseudotype retrovirus which integrates hemagglutinin(HA)of H5N1 avian influenza virus(AIV)isolated from human in Shenzhen.Methods AIV HA gene was amplified bv RT-PCR,then it was ligated with pGEM-T vector,and identified by restriction enzyme digestion and sequenced.HA gene was cloned into CMV/R vector at the site of Sal Ⅰ and BamH Ⅰ.pHR-Luc,pCMV&8.2 and CMV-HA were co-transfected into 293T cell by co-precipitation with calcium phosphate.The pseudotype virus supernatant was harvested 72 h post-transfection and ultracentrifugation,and the HA and P24 expression on the surface of pseudotype virus was analyzed by western blot.Meanwhile.the infection activity of HIV-HA pseudotype virus was identified in different kinds of cell lines,including MDCK,HeLa,CHO and 293T.Results A/Shenzhen/406H/06 belonged to subclade2.3 with open reading frame(ORF)of HA gene encoded 567 amino acides,whose accession number was EF137706 in GenBank.HA gene was cloned into CMV/R successfully.After co-transfection of above vectors,it revealed that HA protein could integrate pseudotype virus by western blot,and precursor protein HA0 could cleavage into HA1 and HA2 with biological activity.Finally.HIV-HA pseudotype virus could infect 4 kinds of cell lines,which indicated its property of infectivity and catholicity.Conclusion The pseudotype retrnvirns wassuccessfully constructed,which can integrate HA protein of A/Shenzhen/406H/06 and had property of infectivity.It call be used in the further research,including selection of neutralizing antibodies and epitope analysis.
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<p><b>OBJECTIVE</b>To explore a fabricating method of incisor all-ceramic cantilevered resin-bonded fixed partial denture (RBFPD) and evaluate their clinical performances.</p><p><b>METHODS</b>All-ceramic cantilevered RBFPDs were made to replace 1 missing incisor in each of 10 patients. The RBFPDs with lingual surface retainer were made of Empress II ceramic materials and luted with resin cement. The prostheses were examined at 2 weeks, 1 year and 2 years after denture insertion. Examining criteria included: fracture of bridge, marginal adaptation, secondary caries, and color match.</p><p><b>RESULTS</b>The average follow-up period was 14. 7 months. No framework fracture and retainer debonding were observed and marginal adaptation was evaluated as good. Seven RBFPDs matched the color of adjacent teeth, and other three did not match well but within the acceptable range of tooth shade difference.</p><p><b>CONCLUSIONS</b>The results of this preliminary study suggest that the clinical performances of all-ceramic cantilevered RBFPDs are good in short-term evaluation, but their long-term success needs to be further investigated.</p>
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Adult , Female , Humans , Male , Young Adult , Denture, Partial, Fixed , Denture, Partial, Fixed, Resin-Bonded , Follow-Up Studies , IncisorABSTRACT
It is suggested in this paper that the integrated Chinese and Western medicine (ICWM) study on infectious disease should be emphasized on fields of viral hepatitis, AIDS, bacillary phthisis and newly discovered infectious diseases, etc. clinically and theoretically, to unfold the studies of immune regulation on infectious diseases, treatment on bacteria and drug-resistant virus strain, and to promote functional and histological recovery, etc. in patients. Focusing on the weak links and shortages of modern medicine, the valuable experiences and theoretical advantages of TCM in clinical trials and mechanism studies should be brought to light so as to bring Chinese medicine into the frame of modern medicine and to contribute to the world medicine.
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Humans , Anti-Bacterial Agents , Therapeutic Uses , Communicable Diseases , Drug Therapy , Drug Therapy, Combination , Drugs, Chinese Herbal , Therapeutic Uses , Medicine, Chinese Traditional , PhytotherapyABSTRACT
<p><b>BACKGROUND</b>The palate is differently regulated and developed along the anterior-posterior axis. The Bmp signal pathway plays a crucial role in palatogenesis. Conditioned-inactivation of Bmp type I receptor Alk2 or Alk3 in the neural crest or craniofacial region leads to palatal cleft in mice. However, how different Bmp members are involved in palatogenesis remains to be elucidated. In the present study, mRNA expression patterns of Bmp2, Bmp3 and Bmp4 in the developing anterior and posterior palates were examined and compared, focusing on the fusion stage.</p><p><b>METHODS</b>To detect the expression of Bmp mRNA, antisense riboprobes were synthesized by in vitro transcription. Radioactive in situ hybridization was performed on sagital and coronal sections of mice head from E13 to E18.</p><p><b>RESULTS</b>The expression of these Bmps were developmentally regulated in the anterior and posterior palates prior to, during and after palatal fusion. During palatal fusion, Bmp4 expression shifted from the anterior to the posterior palate, Bmp2 was highly expressed in both the anterior and posterior palates in this process, whereas Bmp3 was only localized in the posterior palate. They showed generally non-overlapping pattern in their expression domains. Thereafter, their expression was detected in both the anterior and posterior palates regulating osteogenesis and myogenesis respectively.</p><p><b>CONCLUSIONS</b>Bmp signalling is involved in palatogenesis in multiple stages and has multiple roles in regulating anterior and posterior palatal development. Disturbances of Bmp signalling during palatogenesis might be a possible mechanism of cleft palate.</p>
Subject(s)
Animals , Female , Mice , Bone Morphogenetic Protein 2 , Bone Morphogenetic Protein 3 , Bone Morphogenetic Protein 4 , Bone Morphogenetic Proteins , Genetics , Gene Expression Regulation, Developmental , Palate , Embryology , Metabolism , RNA, Messenger , Signal Transduction , Transforming Growth Factor beta , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To observe the therapeutic effect of Zhenggan Recipe (ZGR) in treating liver cirrhosis with abnormal Alpha-fetoprotein (AFP), and to explore its mechanism.</p><p><b>METHODS</b>Sixty-four patients of liver cirrhosis with abnormal AFP were randomly divided into the treated group and the control group. All the patients received conventional combined Western medicinal treatment. Besides, additional ZGR was given to the patients in the treated group, one dose every day for orally taken in two times, 6 months as a therapeutic course. Changes in liver function (ALT, GGT, TBIL and ALB), AFP, T-lymphocyte subsets (CD4, CD4/CD8) and iconological examination were observed in the two groups before and after treatment to compared the efficacy.</p><p><b>RESULTS</b>ZGR could effectively lower the AFP level (P < 0.01), and improve the levels of CD4 and CD4/CD8 (P < 0.01 or P < 0.05).</p><p><b>CONCLUSION</b>ZGR can effectively lower the level of AFP, its mechanism might be related to the improving of cellular immune function of the patients.</p>
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Adult , Aged , Female , Humans , Male , Middle Aged , CD4-CD8 Ratio , Drugs, Chinese Herbal , Therapeutic Uses , Liver Cirrhosis , Blood , Drug Therapy , Allergy and Immunology , Phytotherapy , alpha-Fetoproteins , MetabolismABSTRACT
PURPOSE AND METHODS In order to investigate the effects of Zhaolian on anti tumor promotion,we studied its influence on liver precancerous lesion in rats induced by DEN.RESULTS (1)Liver function of rats was protected ( P