To explore the regularity for the multiplication of avian influenza virus subtype H9N2 in large-scale microcarrier-based MDCK cell culture system, and to determine the optimal proliferation conditions. H9N2 subtype of avian influenza virus was inoculated into the MDCK cell growing on 24 well plate, and the HA titers of virus at different time were detected in the conditions of different infectious doses,different concentrations of TPCK- trypsin and different pH. The optimal conditions were determined. Then the H9N2 subtype avian influenza virus was grown in microcarrier-based MDCK cell in 250mL and 5L roller bottles. It was demonstrated that high viruse yield with a hemagglutination unit of 9 log2(1:512) could be obtained under the optimal conditions of multiplication . The result indicated the H9N2 subtype avian influenza virus could be produced in microcarrier-based MDCK cell in a large-scale culture system with a high virus yield and demonstrates the feasibility of the development of mammalian cell-based in influenza vaccine in microcarrier culture systems.

OBJECTIVETo study the effect of electro-acupuncture (EA) at Neiguan (PC6) and Lieque (LU7) on the expression of protein kinases in cardiomyocytes of myocardial ischemia (MI) rats.

METHODSHealthy male SD rats were randomly divided into the control group, the model group, the Neiguan point group, the Lieque point group, and the non-meridian non-acupoint group, 10 in each group by random digit table. The MI rat model was established by injecting isoprenaline hydrochloride (85 mg/kg). EA at Neiguan (PC6), Lieque (LU7), and non-meridian non-acupoint were respectively performed. Changes of the expression of protein kinases [such as protein kinase A (PKA), protein kinase C (PKC), protein kinase G (PKG)] in rat cardiomyocytes were observed using Western blot.

RESULTSCompared with the control group, expression levels of PKA, PKC, and PKG increased obviously in the model group (P < 0.01). Compared with the model group, expression levels of PKA, PKC, and PKG decreased in the Neiguan point group and the Lieque point group (P < 0.01, P < 0.05). Expression levels of PKA decreased in the non-meridian non-acupoint group (P < 0.01). Compared with the Neiguan point group, expression levels of PKA, PKC, and PKG increased in the non-meridian non-acupoint group and the Lieque point group (P < 0.01, P < 0.05). Compared with the Lieque point group, expression levels of PKA, PKC, and PKG increased in the non-meridian non-acupoint group (P < 0.01, P < 0.05).

CONCLUSIONEA at Neiguan (PC6) and Lieque (LU7) could decrease protein expression levels of PKA, PKC and PKG in rat myocardial cells, and the effect of acupuncture at Neiguan (PC6) was better than that obtained by EA at Lieque (LU7).

Acupuncture Points ; Acupuncture Therapy ; Animals ; Coronary Artery Disease ; Cyclic AMP-Dependent Protein Kinases ; Electroacupuncture ; Male ; Meridians ; Myocardial Ischemia ; metabolism ; therapy ; Myocytes, Cardiac ; metabolism ; Plant Extracts ; Protein Kinases ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the effects of curcumin on the expressions of TNF-alpha, IL-6 and IL-8 in rats with chronic nonbacterial prostatitis.

METHODSSixty healthy adult male SD rats with the body weight of 200 -220 g were equally and randomly divided into a normal control, a positive control, a model, an oral curcumin and an intraperitoneal curcumin group. The rat models of chronic nonbacterial prostatitis were made by hypodermic injection of estradiol benzoate at the dose of 0.25 mg/(kg x d) for 30 days after castration, and then treated with curcumin at 200 mg/(kg x d) by gavage or intraperitoneal injection. The positive controls received oral celebrex at 250 mg/(kg x d), while the normal control and model groups were given saline by gavage. After a week of treatment, the levels of TNF-alpha, IL-6 and IL-8 in the serum and prostate tissues of the rats were detected by ELISA assay.

RESULTSThe levels of TNF-alpha and IL-8 in the serum and prostate tissues were significantly lower in the intraperitoneal curcumin than in the positive control group (P < 0.05), but the expression of IL-6 showed no significant difference between the two groups (P > 0.01).

CONCLUSIONCurcumin is efficacious for chronic nonbacterial prostatitis in rats, and the action mechanism may be associated with its decreasing effect on the proinflammatory cytokines IL-8 and TNF-alpha in the blood and tissues.

Animals ; Chronic Disease ; Curcumin ; pharmacology ; therapeutic use ; Drugs, Chinese Herbal ; therapeutic use ; Interleukin-6 ; metabolism ; Interleukin-8 ; metabolism ; Male ; Phytotherapy ; Prostatitis ; drug therapy ; metabolism ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVE: To explore the genetic basis of a Chinese pedigree affected with Dyggve-Melchior-Clausen syndrome. METHODS: Whole exome sequencing and Sanger sequencing were carried out to detect potential pathogenic variants associated with the syndrome. The function of candidate variant was verified by Western blotting. RESULTS: A novel homozygous variant, c.1222delG of the DYM gene was detected in the two affected siblings, for which both parents were heterozygous carriers. The variant has caused replacement of Asp by Met at amino acid 408 and generate a premature stop codon p.Asp408Metfs*10. Western blotting confirmed that the variant can result in degradation of the mutant DYM protein, suggesting that it is a loss of function variant. CONCLUSION The homozygous c.1222delG frameshift variant of the DYM probably underlay the Dyggve-Melchior-Clausen syndrome in the two affected siblings. Above findings has enabled clinical diagnosis and genetic counseling for the family.
China ; Dwarfism/genetics* ; Humans ; Intellectual Disability ; Osteochondrodysplasias/genetics* ; Pedigree

Adult ; China ; epidemiology ; Female ; Humans ; Low Back Pain ; epidemiology ; etiology ; Occupational Diseases ; epidemiology ; etiology

Adolescent ; Bone Neoplasms ; diagnosis ; Diagnostic Errors ; Humans ; Lymphoma ; diagnosis ; Male

OBJECTIVETo evaluate the curative effects of manipulative reduction for children's type III humeral supracondylar fracture and the preventions of ischemic contracture of forearm in the early period.

METHODSFrom September 2008 to September 2011, 38 patients with humeral supracondylar fractures were treated with manipulative reduction and plaster stabilization, including 20 males and 18 females with an average age of 7.5 years (ranged, 2 to 13 years); the average time from injury to visit was 1.8 days(ranged,0.5 h to 6 d). There were 21 cases in straighten-ulnar deviation type and 17 cases in straighten-radial deviation type, 1 case in flexion type,all of them without vascular nerve injury. It was important to process swelling correctly in early stage of fracture. To decide fixed position according to the original displacement, and make a regular X-ray review, if found another displacement to correct it in 1-2 weeks after injury in time. Dismantle the plaster on the basis of bone healing and guide the functional exercise of elbow joint. According to Dodgt standard to evaluate clinical effects.

RESULTSAll patients were followed up from 3 months to 1 year with an average of 7 months. All fractures healed. According to Dodgt standard, 14 patients got an excellent results, 19 good, 4 fair and 1 poor. The excellent and good rate was 86.84%.

CONCLUSIONIt can obtain satisfactory clinical effects to treat humeral supracondylar fracture in children with closed manipulative reduction and plaster stabilization, while without vascular nerve injury. Early correct processing swelling and paying attention to gypsous angle can effectively prevent the ischemic contracture of forearm.

Adolescent ; Casts, Surgical ; Child ; Child, Preschool ; Female ; Forearm ; blood supply ; Humans ; Humeral Fractures ; therapy ; Ischemic Contracture ; prevention & control ; Male ; Manipulation, Orthopedic

BACKGROUNDObstructive sleep apnea syndrome (OSAS) is an important risk factor for cardiovascular diseases. Chronic intermittent hypoxia (CIH) is considered to be one of the most important causes of cardiovascular diseases in OSA patients. This repeated hypoxia and reoxygenation cycle is similar to hypoxia-reperfusion injury, which initiates oxidative stress. In this study, we observed cardiocytes injury induced by CIH and the effect of N-acetylcysteine (NAC).

METHODSThirty ICR mice were randomly assigned to 3 groups: control, CIH and NAC (CIH + NAC) groups. Malondialdehyde (MDA) and superoxide dismutase (SOD) of cardiocyte homogenates were measured. Serum lipids were measured by an instrument method. Serum cardiac troponin I (cTnI) was detected by enzyme-linked immunosorbent assays (ELISA). Myocardium pathological sections were observed.

RESULTS(1) The SOD activity and MDA concentration of cardiocyte homogenates in the CIH group were significantly higher than in other groups (P < 0.005). The MDA concentration of the NAC group was lower than that of the control group (P < 0.01). (2) The serum cTnI concentration of the CIH and NAC groups was significantly higher than that of the control group (P < 0.01). (3) Serum triglyceride levels in the NAC group were lower than in the other groups (P < 0.01), while there were no significant differences in low density lipoprotein and high density lipoprotein among the three groups. (4) The degeneration of myocardium, transverse striation blurred, and fabric effusion were observed in tissue sections in the CIH and NAC groups. However, normal tissue was found in the control group.

CONCLUSIONThe oxidative stress induced by CIH can injure cardiocytes and the injury effect can be partially inhibited by NAC.

Acetylcysteine ; pharmacology ; Animals ; Free Radical Scavengers ; pharmacology ; Heart ; drug effects ; Hypoxia ; physiopathology ; Malondialdehyde ; metabolism ; Mice ; Mice, Inbred ICR ; Myocardium ; metabolism ; pathology ; Oxidative Stress ; physiology ; Random Allocation ; Superoxide Dismutase ; metabolism

OBJECTIVETo study the immuno-tolerance mechanism of the third-party bone marrow-derived mesenchymal stem cells (BMSCs) in the allogeneic transplantation.

METHODSForty female C57BL/6 mice and forty male BALB/C mice were respectively used as donors and recipients in skin allogenic graft model. Forty male BALB/C mice were divided randomly into 4 groups: blank control group, CP group, BMSCs group , CP + BMSCs group, with 10 mice in each group. Before skin graft, high-dose abdominal injection of cyclophosphamide (200 mg/kg, 2 d, q. d.) was performed in recipient mice in CP and CP + BMSCs groups. On the transplantation day, a bonus of 2 x 10(6) BMSCs from the SD rat (SD-BMSCs) were injected through the tail vein in the BMSCs and CP + BMSCs groups. The observation and HE staining of skin grafts were used. The expressions of CD29, CD34, CD45 and CD90 of cells were analyzed by using flow cytometry in order to identify BMSCs. The CD4+, CD25+, Foxp3 and Treg cells of spleen were detected by flow cytometry. Cytokine in peripheral blood of recipient mice were measured by ELISA, including TGF-beta, IL-10 and IFN-gamma. T cells were co-cultured with 60Co-irradiated bone marrow MSCs from different individuals. The proliferative activity of T cells were evaluated with MTT assay.

RESULTSThe skin graft survival time was significantly prolonged in the CP + BMSCs group, as compared with that in the blank control group, the CP group, the BMSCs group, respectively. Cells cultured by whole bone marrow adherent cultivation showed CD29 (99.7%), CD44+ (96.7%), CD34- (1.6%), CD45- (1.3%). Compared with the control group and CP group, the ratio of the CD4+, CD25+, Foxp3+ and Treg cells significantly increased in the SD-BMSCs group and CP + BMSCs group (P < 0.05). Analysis of peripheral blood by ELISA showed significant high level of TGF-beta, IL-10 and low level of IFN-gamma in BMSCs group and CP group,compared with that in control group. When co-cultured with BMSCs from different individuals, T- lymphocytes proliferation decreased apparently in SD-BMSCs group and C57-BMSCs group (P < 0.05), but there was no significant difference between SD-BMSCs group and C57-BMSCs group (P > 0.05).

CONCLUSIONSThe immunotolerance mechanism of the third-party bone marrow-derived mesenchymal stem cells in the allogeneic transplantation might be associated with its effect on the proliferation of Treg cells and increasing expression of TGF-beta and IL-10, decreasing expression of IFN-gamma.

Animals ; Bone Marrow Cells ; immunology ; Female ; Immune Tolerance ; Interferon-gamma ; immunology ; Interleukin-10 ; immunology ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; immunology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Rats ; Rats, Sprague-Dawley ; Skin Transplantation ; T-Lymphocytes, Regulatory ; immunology ; Transforming Growth Factor beta ; immunology ; Transplantation, Homologous

Transforming technology for semi-synthesized isocorydione from the natural product ofisocorydine was studied. The factors affecting on the reaction yield were investigated. UV spectrophotometry was used to indicate the semi-synthesized yield of isocorydione. The optimum reaction conditions were determined as following: reacting for 12 h in the solution of sodium dihydrogen phosphate at pH 10, the temperature was 25 degrees C and the ratio of isocorydine to Fremy's radical was 1 : 2. Under the optimum conditions, the yield could reach up to 50.0%. The molecular structure of isocorydione was elucidated by X-ray single-crystal diffraction analysis for the first time.
Antineoplastic Agents, Phytogenic ; chemical synthesis ; chemistry ; isolation & purification ; Aporphines ; chemical synthesis ; chemistry ; isolation & purification ; Crystallography, X-Ray ; Hydrogen-Ion Concentration ; Molecular Structure ; Nitroso Compounds ; Oxidation-Reduction ; Papaveraceae ; chemistry ; Plants, Medicinal ; chemistry ; Spectrophotometry, Ultraviolet ; Temperature ; X-Ray Diffraction