Tissue samples of Fabricius' bursa collected from Nanning, Yulin, Beihai and Wuzhou in the provinces of Guangxi in China during the years of 2000-2007, were detected by a established reverse transcriptase polymerase chain reaction (RT-PCR) technique for IBDV. Viral isolation was performed on the positive samples by chicken embryo inoculation via chorio-allantoic membrane (CAM). Results showed that 27 isolates of IBDV were obtained. A set of primers were designed to amplify the vVP2 of 27 isolates by RT-PCR and the PCR products were sequenced. The sequences of all the isolates and reference viruses were analyzed and compared, and their phylogenetic trees were constructed based on the nucleotide sequences. The results indicated that isolate BH11, TZ(3), 050222, YL051, NN0603, NN0611and QX0602 etc, altogether 17 isolates, which accounted for 62.96 percent of total isolates, were identified to be very virulent IBDV (vvIBDV) and have the highest homology to vvIBDV reference strains. In the phylogenetic analysis, they are divided into 3 groups and have a long distance to commonly used vaccine stains. Isolate NN040124 and YL052 were identified as intermediate-plus virulent strains and showed a highest homology to classical strains of 52-70 and STC. 8 isolates of YLZF2, 040131 etc were identified as attenuated vaccine strains and showed a highest homology to classical strain of CU1. The results from the study demonstrated that the viruses prevailing in chickens in these 4 regions in Guangxi province in the recently 7 years were vvIBDV and their origins were complex. The antigenicity of some isolates may have been drifted.
Amino Acid Sequence ; Animals ; Birnaviridae Infections ; epidemiology ; veterinary ; virology ; Chickens ; China ; epidemiology ; Infectious bursal disease virus ; chemistry ; classification ; genetics ; isolation & purification ; Molecular Epidemiology ; Molecular Sequence Data ; Phylogeny ; Poultry Diseases ; epidemiology ; virology ; Sequence Alignment ; Viral Proteins ; chemistry ; genetics

Adolescent ; Child ; Child, Preschool ; Cytomegalovirus Infections ; complications ; Female ; Humans ; Kidney Diseases ; etiology ; Kidney Glomerulus ; pathology ; Male

HuaFu Shengji is the primary traditional Chinese medicine (TCM) therapy for treating chronic skin ulcer. The high activities of the protein enzyme in the wound fluids is one of the main cause of healing delay. In order to investigate the effect of TCM Zhuhong ointment for promoting wound healing. This research focused on its influence on matrix metalloproteinase (MMP) activities in wound fluids with TCM Yang syndromes, directly on the activated MMP-1,2 activities in vitro and on MMP-1,-2,-9 production by HSF. 8 wound fluid samples were collected, which were diagnosed Yang Syndromes in TCM. Wound fluid activities of MMP-2 and MMP-9 were measured by gelatin zymogram assay. MMP-1 and MMP-2 activities in vitro were measured by substrate cleavage. CCK-8 was used to observe the toxicity of Zhuhong ointment on HSF. MMP-1,-2,-9 production by HSF were detected by confocal microscope. Zhuhong ointment from 1 to 25 g x L(-1) obviously inhibited MMP-2 activity in wound fluid. When Zhuhong ointment was over 5 g x L(-1), it showed significantly inhibitory effect on wound fluid MMP-9 activity. In vitro study, when the mercury concentration was 320 mg x L(-1), Zhuhong ointment solution directly inhibited both MMP-1 activity and MMP-2. But mercury concentration from 0.51-2.56 mg x L(-1), it could activate MMP-1 activity, and from 0.51-64 mg x L(-1), activate MMP-2 activity instead. The mercury concentration when Zhuhong ointment saturated in DMEM was 39.6 mg x L(-1). When the mercury concentration was over 1.23 mg x L(-1), Zhuhong ointment showed toxicity to HSF. At 1.23, 0.62, 0.31 mg x L(-1) of mercury concentration, it increased MMP-1 expression by HSF, and at 1.23, 0.62 mg x L(-1), decreased MMP-2 expression. However, at 1.23, 0.62, 0.31 mg x L(-1), it decreased MMP-9 expression. At higher concentration, Zhuhong ointment can inhibit MMP-2, MMP-9 activities in wound fluid with dose-dependent way and show a direct inhibitory effect on activated MMP-1 and MMP-2 in vitro. But at a lower concentration, it showed two-way adjustment, with increased MMP-1, MMP-2 activities and its expression by HSF and decreased MMP-9 activity.
Body Fluids ; enzymology ; Cells, Cultured ; Dermatitis ; drug therapy ; enzymology ; physiopathology ; Drugs, Chinese Herbal ; pharmacology ; Fibroblasts ; drug effects ; enzymology ; physiology ; Humans ; Matrix Metalloproteinase 1 ; metabolism ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Wound Healing ; drug effects

Objective To assess the changes and the leading cause of deaths for children under 5 years old,in China,during 2000-2010,with the aim of evaluation on the progress in achieving the relative goal set by "National Program of Action for Child Development in China (2001-2010)",and understanding the related challenges.Methods Data used in this study were collected from the population-based National Maternal and Child' s Health Surveillance Network of China.Infant Mortality Rate (IMR),Under-5-mortality rate (U5MR) and the leading cause of deaths for under-5 children were analyzed.Results Nationwide IMR and U5MR in 2010 dropped by 59.3％and 58.7％ respectively,compared to that in 2000.Decreases by 50.8％ and 47.1％ in IMR and U5MR were observed in urban areas,and 56.5％ and 56.0％ in rural areas during this period.Compared with data from 2000,the leading causes-specific U5MR in 2010 had significantly declined.The top 5 leading causes of death in 2010 were premature birth/low birth weight,pneumonia,birth asphyxia,congenital heart disease and accidental suffocation,but were different in urban and rural areas.In 2010,both IMR and U5MR from the rural areas were 2.8-folds than that of the urban areas.In addition,IMRs in the Middle and Western parts of China were 1.5 and 2.3-folds respectively of that in the East,and U5MR in Middle and West was 1.5 and 2.2-folds respectively of that in East.Conclusion IMR,U5MR and the leading causes specific mortality rate in China declined remarkably from 2000 to 2010,and the goal set by "National Program of Achon for Child Development in China (2001-2010)" had been successfully achieved.However,the disparity on child' s health in regions and in urban or rural areas,still remained a challenge.

AIM To establish an HPLC method for the simultaneous content determination of four constituents in Liujing Toutong Tablets (Angelicae dahuricae Radix,Magnoliae Flos,Ligustici Rhizoma et Radix,etc.).METHODS The analysis of 30％ ethanol extract of this drug was performed on a 35 ℃ thermostatic Waters C18 column (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of methanol-4％ acetic acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 320 nm.RESULTS Puerarin,ferulic acid,imperatorin and isoimperatorin showed good linear relationships within the ranges of 60.6-303 μg/mL (r=0.999 9),1.59-7.95 μg/mL (r =0.999 9),1.57-7.85 μg/mL (r =0.999 9) and 0.752 5-3.762 5 μg/mL (r =0.999 7),whose average recoveries (RSDs) were 97.75％ (1.7％),97.68％ (2.3％),97.94％ (1.0％) and 98.29％ (1.6％),respectively.CONCLUSION This stable and reliable method can be used for the quality control of Liujing Toutong Tablets.

Adult ; Cyclosporine ; therapeutic use ; Female ; Humans ; Immunosuppressive Agents ; therapeutic use ; Red-Cell Aplasia, Pure ; drug therapy ; Vidarabine ; analogs & derivatives ; therapeutic use

This article aims to provide a good experimental method for the study of drug treatment of ulcerative colitis. According to the characteristics of ulcerative colitis's clinical symptoms, common ulcerative colitis animal models were analyzed. Based on the characteristics of clinical symptoms of traditional Chinese medicine and Western medicine for ulcerative colitis disease, the existing commonly used animal models of ulcerative colitis were analyzed to summarize the current matching degree, advantages and disadvantages of the exi-sting animal models of ulcerative colitis and clinical symptoms. At present, studies on ulcerative colitis mainly adopt four types of induction modeling methods, such as immunization, chemical stimulation, compound method and gene model. There are many reported methods of colitis modeling, but no model can reflect the characteristics of clinical symptoms of ulcerative colitis treated with Western or Chinese medicine. This article summarizes the characteristics, clinically relevant symptoms and applicable scope of immunization, chemical stimulation, compound method, and gene model, so as to provide a reliable animal model for subsequent studies of prevention and treatment of colitis.
Animals ; Charadriiformes ; China ; Colitis, Ulcerative/genetics* ; Disease Models, Animal ; Medicine ; Medicine, Chinese Traditional

OBJECTIVETo compare the clinical outcomes between unrelated donor hematopoietic stem cell transplantation (URD-HSCT) and HLA-haploidentical (Hi)-HSCT.

METHODSTwenty-five patients with hematologic malignancies received URD-HSCT and thirty patients received Hi-HSCT. The conditioning regimen consisted of modified BUCY or modified total body irradiation (TBI) plus CY. Acute graft-versus-host disease (aGVHD) prophylaxis consisted of cyclosporin ( CsA), short-term methotrexate (MTX), mycophenolate mofetil (MMF), or the combination of CsA, MTX and MMF plus antithymocyte globulin (ATG) or antilymphocyte globulin (ALG), or the combination of CsA, MTX, MMF, ATG/ ALG and CD25 monoclonal antibody.

RESULTSAll patients in the URD-HSCT group and 29 patients in the Hi-HSCT group were engrafted successfully. The median follow-up duration was 7 (2 -59) months for URD-HSCT group and 7.3 (1 - 35) months for Hi-HSCT group. The 3-year probabilities of disease-free survival (DFS) for URD-HSCT and Hi-HSCT group were (54.1 +/- 11.9)% and (43.1 +/- 9.1)%, respectively (P =0.13). Grade III - IV aGVHD occurred in 10 patients in URD-HSCT group and 11 in Hi-HSCT group (the cumulative incidence 40.0% vs 37.9%, P > 0.05), respectively. Ten patients (40.0%) died of transplantation-related mortality (TRM) in URD-HSCT group and 17 (56.7%) in Hi-HSCT group (P >0. 5). Two patients relapsed in each group (the rate of relapse 8.0% vs 6.0%, P >0.05). The primary causes of death included severe aGVHD with infection,severe pulmonary infection and relapse.

CONCLUSIONBoth URD-HSCT and Hi-HSCT are effective and curable treatment for refractory or high-risk hematologic malignancies. The optimal donor should be chose individually. The severe aGVHD and consequent infection are still the main cause of TRM.

Adolescent ; Adult ; Child ; Female ; Follow-Up Studies ; Graft vs Host Disease ; prevention & control ; Hematologic Neoplasms ; therapy ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Male ; Middle Aged ; Tissue Donors ; Transplantation Conditioning ; Treatment Outcome ; Young Adult

OBJECTIVETo explore the possible role of CD28/CTLA-4 co-stimulators in immune pathophysiology of acquired aplastic anemia(AA).

METHODSBy FACS, the percentages of CD28, CTLA-4 expressing CD3+ CD4+ T cells and the level of Th1, Th2 in bone marrow were detected in 23 AA patients at active phase, 10 at recovery phase and 15 normal controls. The relationship between the co-stimulators, Th1, Th2, and absolute neutrophil counts (ANC) was evaluated.

RESULTS(1) The percentage of CD28 and CTLA-4 expressing CD3+ CD4+ T cells in bone marrow, and CD28+/CTLA-4+ ratios were (31.40 +/- 10.83)%, (2.45 +/- 1.30)% , and 17.02 +/- 13.44 in normal controls respectively, (39.84 +/- 10.89)%, (1.43 +/- 0.67)%, and 43.04 +/- 37.61 in AA at active phase, respectively, (22 +/- 9.08)%, (3.46 +/- 2.26)%, and 10.49 +/- 7.8 in AA at recovery phase, respectively. The percentage of CD28 and CD28+/CTLA-4+ ratio were significantly higher, while CTLA-4 were lower in active phase AA patients than in normal controls (P < 0.05). These values in recovery phase AA were comparable to those in normal controls. (The Th1, Th2, and Th1/Th2 in bone marrow were (4.21 +/- 2.11)%, (1.99 +/- 1.27)%, and 2.46 +/- 1.28 in normal controls respectively, (11.13 +/- 4. 96)%, (2.46 +/- 1.65)%, and 5.20 +/- 1.98 in active phase AA and (5.39 +/- 4.2)9%, (2.53 +/- 2.41)%, and 2.87 +/- 1.43 in recovery phase AA, respectively. The percentage of Th1 and Th1/Th2 ratio were significantly higher in AA patients at active phase than in normal controls (P < 0.05). (3) The CD28+/CTLA-4+ ratio was positively related to the Th1+ /Th2+ ratio (P < 0.05). ANC was negatively related to CD3+ CD4+ CD28+ T cells (P < 0.01), and positively to CD3 + CD4 ' CTLA-4' T cells (P < 0.01) respectively.

CONCLUSION(1) The expression of CD28 was increased while CTLA-4 decreased on the membranes of CD3+ CD4+ T cells in bone marrow of AA patients. (2) The abnormal expression of CD28 costimulator promoted the shift of immune balance to Thl type. (3) The unbalance of CD28+ / CTLA-4+ is important for the immune pathophysiology of AA.

Adolescent ; Adult ; Aged ; Anemia, Aplastic ; immunology ; metabolism ; Antigens, CD ; immunology ; metabolism ; CD28 Antigens ; immunology ; metabolism ; CD4-Positive T-Lymphocytes ; metabolism ; CTLA-4 Antigen ; Child ; Female ; Humans ; Male ; Middle Aged ; Th1 Cells ; immunology ; Th2 Cells ; immunology

OBJECTIVETo explore the activation status of signal pathway of mTOR/S6 in bone marrow (BM) T lymphocytes of refractory/relapsed aplastic anemia patients (AA), and the effects of rapamycin (RAPA) and CTLA-4 immunoglobulin (CTLA-4Ig) on this pathway.

METHODSBM was collected from 13 refractory/relapsed AA patients, 8 newly diagnosed severe AA (SAA) patients and 10 iron deficiency anemia (IDA) (as controls) patients, and cocultured with RAPA and CTLA-4 Ig. The expression of p-mTOR, p-S6 and Interferon gamma (IFN-gamma) in CD3(+)T cells was measured by flow cytometry (FCM).

RESULTS(1) The expression of p-mTOR, p-S6 and IFN-gamma in CD3(+)T cells in refractory/relapsed AA group were significantly higher than those in controls (P < 0.01). (2) The expression of p-mTOR and p-S6 in T cells in newly diagnosed SAA group, was similar to those in controls (P > 0.05), but significantly lower than those in refractory/relapsed AA group (P < 0.01). The expression level of IFN-gamma in T cells were significantly higher than that in controls (P < 0.01). (3) On exposure to RAPA, the levels of p-mTOR, p-S6 and IFN-gamma in T cells in refractory/relapsed AA patients were significantly lower than those before the exposure (all P < 0.05). And so were when exposed to CTLA-4 Ig (all P < 0.01).

CONCLUSION(1) The mTOR/S6 signal pathway is activated in refractory/relapsed AA. (2) The expression of p-mTOR, p-S6 and IFN-gamma in refractory/relapsed AA can be suppressed by RAPA or CTLA-4Ig. (3) The signal pathway of CD28/mTOR/S6/IFN-gamma might take part in immune pathogenesis of refractory/relapsed AA.

Adolescent ; Adult ; Aged ; Anemia, Aplastic ; immunology ; metabolism ; Antigens, CD ; pharmacology ; CTLA-4 Antigen ; Child ; Female ; Humans ; Interferon-gamma ; metabolism ; Male ; Middle Aged ; Ribosomal Protein S6 ; metabolism ; Signal Transduction ; drug effects ; Sirolimus ; pharmacology ; T-Lymphocytes ; immunology ; metabolism ; TOR Serine-Threonine Kinases ; metabolism ; Young Adult