0.05). But the level of HbA1c, 2 hour insulin and C peptide were significantly lower than that in CG (P 0.05). Conclusion Wei li acupuncture therapy instrument can improvement the islet ?-cell function in patients with type 2 diabetes mellitus.

Duck enteritis virus (DEV) is a herpesvirus that causes an acute, contagious and fatal disease. In the present article, the DEV UL4 gene was cloned and sequenced from a vaccine virus. A degenerate oligonucleotide primer for the consensus site of herpesvirus UL3 gene and a specific primer located in UL5 were used in the polymerase chain reaction (PCR) to amplify a DNA product 2 086 bp in size. DNA sequence analysis revealed that a 714 bp open reading frame (ORF) of DEV encoding a 237 amino acid polypeptide is homologous to the family of herpesvirus UL4 proteins and therefore has been characterized as a DEV UL4 gene. Alignment of the DEV UL4 protein sequence with those of other alphaherpesviruses showed that 10 amino acid residues are completely conserved. Phylogenetic tree analysis showed that the seventeen alphaherpesviruses viruses analyzed were classified into four large groups, and the duck enteritis virus branched separately, closely related to the Mardiviruses group comprising Gallid herpesvirus 2 (GaHV-2), Gallid herpesvirus 3 (GaHV-3) and Meleagrid herpesvirus 1 (MeHV-1). The present study showed that the evolutionary relationship of the UL4 protein could be used for classification of alphaherpesviruses.

Objective:To obtain eukaryotic expressing protein of chicken interferon ? (ChIFN-?) and research its anti-virus activity.Methods: Chicken interferon ? mature protein gene was cloned and amplified by reverse transcripition-polymerase chain reaction(RT-PCR) from the total mRNA in the lymphocyte of chicken blood stimulated with ConA for 4～10 hours.The gene was inserted into the expression vector pPICZa-A,which had been cleaved by EcoR I and Xba I.The recombinant vector was linearized by Sac I and transferred into yeast Pichia pastoris,strain X33,anti-virus activity of the recombinant cytokine was detected by the classical experiment cell pathological effect inhibition assay.Results:The result showed that the preparation of recombinant interferon had higher anti-virus activity(10?48 U/ml).Conclusion: The recombinant chicken interferon ? with anti-virus bioactivity has been obtained.

Heat-shock protein (Hsp) 70 potentiates specific immune responses to some antigenic peptides fused to it. Here, the prokaryotic plasmids harboring the envelope glycoprotein E0 gene of classical swine fever virus (CSFV) and/or the Hsp70 gene of Haemophilus parasuis were constructed and expressed in Escherichia coli Rosseta 2(R2). The fusion proteins were then purified. Groups of Balb/c mice were immunized with these fusion proteins, respectively, and sera collected 7 days after the third immunization. Immune effects were determined via an enzyme-linked immunosorbent assay and flow cytometric analyses. E0-Hsp70 fusion protein and E0+Hsp70 mixture significantly improved the titer of E-specific antibody, levels of CD4+ and CD8+ T cells, and release of interferon-γ. These findings suggested that Hsp70 can significantly enhance the immune effects of the envelope glycoprotein E0 of CSFV, thereby laying the foundation of further application in pigs.
Animals ; Antibodies, Viral ; blood ; CD4-Positive T-Lymphocytes ; cytology ; immunology ; CD8-Positive T-Lymphocytes ; cytology ; immunology ; Cell Proliferation ; Classical swine fever virus ; genetics ; Female ; HSP70 Heat-Shock Proteins ; genetics ; immunology ; Haemophilus parasuis ; genetics ; Immunization ; Interferon-gamma ; metabolism ; Mice ; Mice, Inbred BALB C ; Plasmids ; genetics ; Recombinant Fusion Proteins ; genetics ; immunology ; Viral Envelope Proteins ; genetics

Thymopentin (TP5) and bursopentin (BP5) are both immunopotentiators. To explore whether the TP5-BP5 fusion peptide (TBP5) has adjuvant activity or not, we cloned the TBP5 gene and confirmed that the TBP5 gene in a recombinant prokaryotic expression plasmid was successfully expressed in Escherichia coli BL21. TBP5 significantly promoted the proliferation of thymic and splenic lymphocytes of mice. The potential adjuvant activity of the TBP5 was examined in mice by coinjecting TBP5 and H9N2 avian influenza virus (AIV) inactivated vaccine. HI antibody titers, HA antibodies and cytokines levels (IL-4 and IFN-γ) were determined. We found that TBP5 markedly elevated serum HI titers and HA antibody levels, induced the secretion of both IL-4 and IFN-γ cytokines. Furthermore, virus challenge experiments confirmed that TBP5 contributed to inhibition replication of the virus [H9N2 AIV (A/chicken/Jiangsu/NJ07/05)] from mouse lungs. Altogether, these findings suggest that TBP5 may be an effective adjuvant for avian vaccine and that this study provides a reference for further research on new vaccine adjuvants.
Adjuvants, Immunologic ; pharmacology ; Animals ; Antibodies, Viral ; blood ; Cell Proliferation ; drug effects ; Influenza A Virus, H9N2 Subtype ; drug effects ; physiology ; Influenza Vaccines ; immunology ; Interferon-gamma ; immunology ; Interleukin-4 ; immunology ; Lymphocytes ; drug effects ; Mice ; Oligopeptides ; immunology ; Orthomyxoviridae Infections ; drug therapy ; Recombinant Fusion Proteins ; immunology ; Spleen ; cytology ; Thymopentin ; immunology ; Thymus Gland ; cytology ; Vaccines, Inactivated ; immunology ; Virus Replication

To investigate the effects of medicated serum prepared by administration of Jinmaitong (JMT), a compound Chinese herbal medicine, on nicotinamide adenine dinucleotide phosphate (NADPH) oxidase p22-phox subunit and inducible nitric oxide synthase (iNOS) of rat Schwann cells cultured in high-glucose medium.

Objective To detect and analyze the resistance at the stomach meridian acupoints in chronic atrophic gastritis (CAG) patients with spleen-stomach deficiency-cold syndromes, summarize the status of asthenia-sthenia at stomach meridian under this condition, in order to guide the clinical therapy with acupuncture and moxibustion.Methods From 2010 January to 2013 December, 83 CAG patients with spleen-stomach deficiency-cold syndromes were collected from No.306 Hospital of the PLA, the surface resistances on the bilateral stomach meridian stomach acupoints were detected,including Lidui, Neiting, Xiangu,Chongyang, Jiexi, Fenglong, Zusanli, Liangqiu, and the non-acupoints 1cm lateral to the point were set as controls. Zhu auricle holographic detector was adopted to detect the resistance at 20 points (about 0.5 cm2around the acupoints) in each acupoint, and the average value was calculated as the resistance value.Results Theresistances on the leftChongyang andright Chongyang were (9.64 ± 2.03) k? and (9.68 ± 2.02) k?, respectively, both were significantly lower than those on other acupoints, with statistically significant difference (allP<0.01); theresistances on the leftLiangqiu andright Liangqiu were (13.44 ± 2.11) k? and (13.68 ± 2.12) k?, respectively, both were significantly higher than those on other acupoints (all P<0.01).Conclusions The resistance on the stomach meridian stomach acupoints in CAG patients with spleen-stomach deficiency-cold syndromes is different, which may reflect the basic pathogenesis of CAG being the weakness of the spleen and stomach, qi stagnation and blood stasis.

The highly virulent PRRSV isolate strain HN-1/06 was cultivated on Marc-145. To study the viral entry mechanisms, the GP5 gene of PRRSV isolate was amplified by RT-PCR and cloned into pcDNA3.0 to generate the expressing plasmid pcDNA-GP5. pcDNA-GP5 was transfected into 293T by the calcium phosphate precipitation method. Analysis of flow cytometry confirmed that the GP5 proteins were expressed in surface of the 293T cells. Then 293T cells were transfected with pcDNA-GP5, pHIT60 and pHIT111 plasmids to generate pseudotyping virus. The pseudotyping virus supernatant was harvested 48 hours post-transfection and was detected by Western blotting and infection assay. Western blotting indicated that the GP5 glycoproteins were incorporated into the retroviral pseudotyped virus. Infection assay showed that the pseudotyped virus infected 293T and Mark-145 cell. The pseudotyped virus could be used to further study infectious mechanism of PRRSV.
Animals ; Cell Line ; Cloning, Molecular ; Endothelial Cells ; cytology ; metabolism ; virology ; Leukemia Virus, Murine ; genetics ; metabolism ; Mice ; Porcine respiratory and reproductive syndrome virus ; chemistry ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Swine ; Transfection ; Viral Envelope Proteins ; biosynthesis ; genetics ; Virion ; genetics ; metabolism

In order to research immunogenicity of the recombinant rVP2-IL-2 fusion protein, we obtained the rVP2-IL-2 fusion protein using Pichia pastoris expression system, and then evaluated its potential to induce immune responses in chicken. The effect was determined in the form of protective anti-IBDV VP2 titers, antibodies (IgG1 and IgG2a), lymphocyte proliferation, the levels of interferon-gamma and interleukin-4 cytokines, and challenge experiment. Antibody titers and proliferation lymphocyte level suggested that the fusion protein could elicit specific humoral immune and cellular immune responses, antibody sub-type results indicated that the rVP2-IL-2 fusion protein induced secretion both of IgG1 and IgG2a. The seem result elicited from cytokines ELISA test, secretion of both of Th1 (gamma-IFN) and Th2 (IL-4) were induced by the rVP2-IL-2 fusion protein. Challenge experiment result shown that chicken immunized the rVP2-IL-2 fusion protein obtained 85% protection. These results confirm that the fusion protein enhances the protection against IBDV through both humoral and cell-mediated immunity, and thus could serve as a candidate for the development of IBDV subunit vaccine.
Animals ; Antibodies, Viral ; biosynthesis ; blood ; Chickens ; immunology ; Immunoglobulin G ; blood ; Interleukin-2 ; biosynthesis ; genetics ; Pichia ; genetics ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Th1 Cells ; immunology ; Th2 Cells ; immunology ; Vaccines, Subunit ; immunology ; Viral Structural Proteins ; biosynthesis ; genetics ; Viral Vaccines ; immunology

Objective To discuss different effects of low frequency sound waves of different timbres on microcirculation and transcutaneous oxygen partial pressure of Weizhong (BL40) of healthy people;To study the mechanism of somatosensory music therapy. Methods The same frequency (98.00 Hz), different timbres (sounds of guqin, flute, ocarina, bell, and drum were simulated) low frequency sound waves were played near Weizhong acupoint of 30 healthy persons. Laser Doppler flowmetry was used to analyze microcirculation and the changes of transcutaneous oxygen pressure of meridian acupoints, when each timbre was played for 60, 120, 180, 240 and 300 s. Results Sound waves of bell, drum, and flute made point microcirculation and transcutaneous oxygen pressure gradually increase, and the influence of drum>bell>flute;timbre waves of ocarina and guqin made the point microcirculation and transcutaneous oxygen pressure decrease gradually decrease, and the decrease of ocarina was more significant than that of guqin. In the same time point, sound waves of bell made point microcirculation and transcutaneous oxygen pressure increase more than the other sound waves (P<0.01). Conclusion Low frequency sounds of different timbres belong to different properties of five elements. They have different components in frequency spectrum, and can create different effects on acupoints.