0.05).At the eighth week of training and after ceasing the cognitive training for 4 weeks the NCSE scores and the FIM scores were improved in both groups,espeeially in the cognitive training group(P

Background Growth factor-induced proliferation and migration of retinal pigment epithelium (RPE) cells are the major pathological changes of proliferative vitreoretinopathy (PVR).Arsenic trioxide ( As2O3 ) is an active ingredient of Chinese traditional medicines,which has an inhibition on proliferation and migration of tumor cells.However,it is not clear whether As2O3 could inhibit growth factor-induced proliferation and migration of RPE cells. Objective This study was to explore the effects of As2O3 on epidermal growth factor (EGF)-induced proliferation and migration of ARPE-19 cells. Methods RPE cell line (ARPE-19 cells) were cultured.Different concentrations of As2O3(0,0.5,1.0,2.0,5.0,10.0,20.0 μmol/L) were added in the culture plate to treat ARPE-19 cells with or without 10 mg/L EGF in serum-free group for 24 and 48 hours,respectively.The MTT colorimetric assay was used to check the cell viability and evaluate the drug toxicity.The effects of As2O3 on EGF-induced proliferation of ARPE-19 cells were analyzed to get an effective and avirulent concentrations of As2O3.The effects of As2O3 on EGF-induced migration of ARPE-19 cells were observed by scratch-wound assay and the Boyden chamber assay.Results MTT assay showed that the A values were gradually declined with the increase of As2O3 concentrations after As2O3 treatment without EGF for 24 hours and 48 hours ( Fgroup =38.269,P =0.000 ; Ftime =0.874,P =0.358 ).Compared with the control group,no significant differences were seen in the A values of ARPE-19 cells in 0.5-5.0 μmol/L groups (all P＞0.05).Meantime,As2O3 reduced the A values of ARPE-19 cell with 10 mg/L EGF in dose- and time-dependent manner ( Fgroup =152.155,P =0.000 ; Ftime =51.649,P =0.000 ).There were not significant differences in 10 mg/L EGF-induced cell growth after 0.5,1.0,2.0 μmol/L As2O3 was added for 24 and 48 hours ( Fgroup =2.215,P =0.126 ;Ftime =2.230,P =0.155).However,when 5.0-20.0 μmol/L As2O3 added,the A values of 10 mg/L EGF-induced ARPE-19 cells lowed,showing a significant difference in comparison with the control groups ( all P＜0.05),with the cellular inhibiting rate 12％,32％,37％ in 24 hours and 39％,44％ and 53％ in 48 hours.Scratch-wound assay showed that EGF-induced horizontal migration of ARPE-19 cells was slow after 0.5-2.0 μmol/L As2O3 treated,and the same results also appeared in cell lognitudinal migration by Boyden chamber assay,with the inhibitory rates 22％,33％ and 46％ respectively. Conclusions As2O3 is avirulent on ARPE-19 cells within definite concentration range.At ≤ 2.0 μmol/L concentrations,As2O3 dose not affect EGF-induced proliferation of ARPE-19 cells,but it suppresses EGF-induced cell migration.At ≥ 5.0 μmol/L concentrations,As2O3 plays an inhibitory role to EGF-induced proliferation of ARPE-19 cells.

Background Oxidative damage plays an important role in pathogenesis of age-related macular degeneration( AMD ),and its mechanism is the destroy of blood-retinal barrier.Müller cells is a primary component to stabilize the inner barrier of the blood-retina.Researches showed that epidermal growth factor(EGF) can promote the proliferation and migration of animal Müller cells,but less study was found in the effect of EGF on human Müller cells. Objective The present study was to investigate the effects of EGF on the proliferation and migration of human Müller cells and its molecular mechanism. Methods Human Müller cell line MIO-M1 cells were cultured and incubated,and cultured cells were identified using glial fibrillory acidic protein (GFAP),factor Ⅷ,α-smooth muscle actin( α-SMA ),keratin and S-100.Different concentrations of EGF( 0,1,10,30,100 mg/L)was added in freeserum DMEM,and the positive rate of the cells was calculated using 5-bromo-2-deoxyuridine(BrdU) method.The cells were divided into EGF group,H2 O2 group,EGF + H2 O2 group,glucose oxidase ( GO ) group,GO + EGF group,EGF + LY294002+H2O2 group according to the different intervention,and the effects of LY294002 on the proliferation of Müller cells (A590 )were detected by colorimetric assay for cellular growth and survival( MTT assay).The scratch test of Müller cells was used to assess the influence of EGF(0,1,10,30,100 mg/L)on H2 O2-induced damage of human Müller cell.Western blot was used to detect the cell proliferation under the protection of EGF on co-cultured cells using LY294002 and H2O2 and the activation of Akt signal pathways. Results The proliferative rates of the cells were 28.0％,32.9％,39.0％ in 10,30,100 mg/L EGF groups respectively and obviously higher than those in 0,1 mg/L EGF groups (24.5 ％,26.2 ％ ).Under the H2O2 culture,GO culture,respectively,the A570 value of the Müller cell in high concentrations of EGF groups was significantly increased in comparison with lower concentrations EGF groups with the statistical significance among the groups( F=23.582,P=0.000).Compared with EGF+H2O2 group,the A570value of the Müller cells was lowed in EGF+LY294002+H2O2 group.The maximum migration rate of Müller cells was found in 10 mg/L EGF group.Western blot revealed that the presence of H2O2 reinforced the expression of Akt in Müller cells,however,pretreatment with 100 mg/L EGF antagonized the harmful effect of H2O2 on Müller cells.Meanwhile,pretreatment with EGF and LY294002 reduced the expression of Akt in Müller cells. Conclusions EGF can induce the proliferation and migration of human Müller cells with the strongest effect in 10 mg/L.100 mg/L exogenous EGF has a stronger protection to the Müiller cells against H2O2-induced cell damage by activating the PI3KAkt cell survival pathway.

Objective To analyze psychological changes of patients underwent the hematopoietic stem cell collection and explore the importance of health education in order to support the proceeding of collection smoothly.Methods A total of 50 patients underwent the hematopoietic stem cell collection were divided into two groups:study group(n = 25)and control group(n =25).The control group performed the pure pharmacotherapy while the study group performed health education based on the pharmacotherapy.Results Indicators including the coping style of discomfort symptoms and the knowledge of health education in the study group were better than that in the control group.Conclusions Psychological status of patients plays an important role during collecting proceeding of hematopoietic stem cell.The health education may raise patient' s related knowledge,thus assisting the successful collection.

OBJECTIVETo observe epithelial-mesenchymal phenotypes and oxidative stress related protein expressions of the liver cells in a rat model of liver fibrosis induced by bile duct ligation and recanalization.

METHODSTwenty-four male Wistar rats were randomized into 4 groups, including a sham-operated group, two bile duct ligation groups with ligation for 2 and 4 weeks, and a bile duct ligation group with a 2-week ligation followed by a 2-week recanalization. HE staining and Masson staining were used to assess liver fibrosis in the rats, and immunohistochemistry and Western blotting were employed to detect expressions of the epithelial and mesenchymal marker proteins and oxidative stress-related proteins.

RESULTSCompared with the sham-operated group, the rats with bile duct ligation showed obvious liver fibrosis, which worsened as the ligation time extended, accompanied by significantly increased expression of α-SMA, collagen I, NOX(4) and vimetin and reduced E-cadherin expression. Compared with the rats with bile duct ligation for 4 weeks, the rats in bile duct ligation-recanalization group showed obviously lessened liver fibrosis, significantly lowered expressions of NOX(4) and mesenchymal cell maker proteins, and enhanced expressions of epithelial cell marker proteins.

CONCLUSIONBile duct ligation up-regulates mesenchymal phenotype-related proteins and NOX(4) protein expression and down-regulates the expression of epithelial phenotype-related proteins, and these changes can be reversed by subsequent bile duct recanalization.

Actins ; metabolism ; Animals ; Bile Ducts ; surgery ; Cadherins ; metabolism ; Collagen Type I ; metabolism ; Disease Models, Animal ; Epithelial Cells ; cytology ; metabolism ; Hepatocytes ; cytology ; metabolism ; Ligation ; Liver Cirrhosis ; metabolism ; Male ; NADPH Oxidase 4 ; NADPH Oxidases ; metabolism ; Oxidative Stress ; Phenotype ; Rats ; Rats, Wistar ; Vimentin ; metabolism

BACKGROUND:Al ogeneic hematopoietic stem cel transplantation is currently recognized as the first-line therapy for severe aplastic anemia. However, with the popularity of the one-child families, the source of ful y matched hematopoietic stem cel transplantation is limited, so haploidentical hematopoietic stem cel transplantation is favored. OBJECTIVE:To retrospectively compare and analyze the clinical efficacy and safety of haploidentical al ogeneic hematopoietic stem cel transplantation and ful y matched hematopoietic stem cel transplantation for the treatment of severe aplastic anemia. METHODS:Clinical data of 15 patients with severe aplastic anemia (treatment group) who underwent haploidentical al ogeneic hematopoietic stem cel transplantation in the Department of Hematology General Hospital of Beijing Military Region from January 2013 to January 2015 were retrospectively analyzed. Pretreatment regimen was cyclophosphamide, fludarabine, Busulfex, combined with anti-human lymphocyte immune globulin. Donors received granulocyte colony-stimulating factor, and the transplantation method was bone marrow mobilization combined with peripheral blood stem cel transplantation. Combined immunosuppressive agents including cyclosporine A, methotrexate, tacrolimus, were adopted for prevention of graft versus host disease. Another 15 cases of severe aplastic anemia undergoing ful y matched hematopoietic stem cel transplantation served as control group over the same period. Complications and survival of the two groups were statistical y analyzed. RESULTS AND CONCLUSION:By the end of July 2015, the median fol ow-up time of the treatment group was 20.7 months (6-30 months), and hematopoietic reconstruction was achieved in al cases, including four cases of graft versus host disease, five cases of pulmonary infection, three cases of sepsis, and one case died of pulmonary infection, one cases died of sepsis, and two cases died of graft versus host disease. In the control group, the median fol ow-up time was 19.7 months (5-28 months), hematopoietic reconstruction was achieved in al cases. There were three cases of graft versus host disease, four cases of pulmonary infection, one case died of pulmonary infection, and two cases died of graft versus host disease. The total survival rates of the two groups were 73%and 80%respectively, with no significant difference (P=0.67). The haploidentical al ogeneic hematopoietic stem cel transplantation for severe aplastic anemia is safe and effective, and the clinical efficacy is comparable to the ful y matched hematopoietic stem cel transplantation.

Objective To explore the clinical significance of the relationship between the immune function and the pathogenesis of aplastic anemia in children with aplastic anemia(AA),along with the incidence of graft versus host disease (GVHD)by monitoring the changes of T lymphocyte subsets dynamically in +1 ,+3,+6,+1 2 months for blood disease patients after allogeneic hematopoietic stem cell transplantation.Methods Twelve AA patients re-ceived allogeneic hematopoietic stem cell transplantation in Department of Hematology,the Affiliated General Hospital of Beijing Military Region of Anhui Medical University,from January 201 3 to January 201 4,including 4 male and 8 fe-male,with average age of 7.92 years old(3 -1 4 years old)with 5 cases of human leukocyte antigen(HLA)matched and 7 cases of HLA mismatched.The level of T lymphocyte subsets including CD3 +,CD4 +,CD8 +,CD4 +/CD8 +, CD56 +,CD4 +CD25 high +FOXP3 +were monitored with flow cytometry before transplantation and in +1 ,+3,+6,+1 2 months after transplantation dynamically in the peripheral blood.While in the same period the level of T lymphocyte subsets was monitored in 1 2 cases of healthy children at the same period as the healthy control group.Results Fol-lowed up to March 201 5,1 0 cases had abnormal cellular immunity (CD4 +/CD8 + ratio inversion)in the 1 2 AA pa-tients.Compared with the control group,in the AA group,CD3 + was slightly higher,(66.79 ±7.35)% and (62.74 ± 5.58)% respectively(P =0.043),CD4 + was decreased by (33.73 ±7.26)% and (39.54 ±3.46)% respectively (P =0.037),CD8 + was increased by (35.69 ±6.78)% and (25.34 ±4.36)%,respectively (P =0.000),CD4 +/CD8 + decreased by 1 .23 ±0.56 and 1 .78 ±0.34 respectively(P =0.001 )and CD56 + was decreased by (7.46 ± 2.80)% and (1 6.73 ±3.70)% respectively(P =0.000),CD4 +CD25 high +FOXP3 + was decreased by (3.3 ± 1 .5)% and (8.1 ±1 .3)% respectively (P =0.003),whose difference was statistically significant (P <0.05).The lever of CD3 +,CD4 +,CD8 +,CD4 +/CD8 +,CD56 +,CD4 +CD25 high +FOXP3 + had a different degree of recovery after transplantation for all cases and returned to normal in +1 2 months basically.In +1 ,+3,+6,+1 2 months after transplantation,the levels of CD4 +CD25 high +FOXP3 + in GVHD positive group and negative group were (0.4 ± 0.6)% and (1 .6 ±0.7)% respectively,(0.7 ±0.3)% and (2.7 ±0.4)% respectively,(1 .1 ±0.5 )% and (2.9 ±0.7)% respectively,(1 .4 ±0.3)% and (3.6 ±0.2)% respectively,which had statistical significance (P <0.05).Conclusions There was abnormal cell immune function in some cases with AA.After transplantation,the level of CD4 +CD25 high +FOXP3 + is closely related to the acute GVHD,which can be used to predict the occurrence of GVHD.

Veregen™ and Fulyzaq are the first two botanical drug products that were approved by the Food and Drug Administration (FDA) to market in the US in recent years. Additional herbal medicines, including Compound Danshen Dripping Pills, Fuzheng Huayu Tablets, Xuezhikang Capsule, Guizhi Fuling Capsule, Kanglaite Capsule and Kanglaite Injection, have filed the investigational new drug (IND) application to the FDA and are in phase II or phase III clinical development. In order to gain better understanding of the process of botanical drug approval in the US, this article examines the aforementioned drugs by looking at their composition, indication, prior clinical experience and clinical development process, and summarizes key features that enabled IND filing and marketing approval by the FDA.
China ; Drug Approval ; legislation & jurisprudence ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Nonprescription Drugs ; therapeutic use ; United States ; United States Food and Drug Administration

This study was aimed to investigate the methylation status of WT1 gene in leukemia cell lines and its relation with expression of WT1 gene. The WT1 gene was silenced by DNA methylation or histone deacetylation, and the expression of WT1 gene was induced by using HDAC inhibitor and/or demethylation agent of DNA. Some leukemia cell lines (U937, HL-60, K562, KG1) were detected by RT-PCR, MS-PCR, restriction analysis, and DNA sequencing. U937 leukemic cells without WT1 mRNA expression were incubated with HDAC inhibitor Trichostatin A (TSA) and/or demethylation agent decitabine. The results showed that the U937 cells did not express WT1 gene, but HL-60, K562 and KG1 cells highly expressed WT1 gene; WT1 gene was unmethylated in HL-60 cells, but methylated in K562 and U937 cells. WT1 expression could be reactivated by co-incubation with TSA and decitabine, but not was observed by using single drug. It is concluded that WT1 promoter is methylated in some leukemia cells, however, the methylation can not affect its expression. DNA methylation and deacetylation of histones are synergistic to inhibit the expression of WT1 in leukemic U937 cells, the combination of TSA with decitabine can induce expression of WT1 gene.
Azacitidine ; analogs & derivatives ; pharmacology ; DNA Methylation ; Gene Silencing ; HL-60 Cells ; Histones ; metabolism ; Humans ; Hydroxamic Acids ; pharmacology ; K562 Cells ; Promoter Regions, Genetic ; U937 Cells ; WT1 Proteins ; genetics

OBJECTIVETo analyse the WT1 expression and its DNA methylation status of its promoter domain.

METHODThe expression of WT1 gene and its DNA methylation status were assayed in leukemia cell lines and normal peripheral blood mononuclear cells (PBMNC) by RT-PCR and MS-PCR.

RESULTSWT1 was overexpressed in HL60, K562 and KG1 leukemia cell lines, but not in U937 and PBMNC. Methylation of WT1 promoter was not observed in HL60 cells.

CONCLUSIONDNA methylation of WT1 gene promotor did not inhibit its expression. Other mechanisms may appear to regulate the WT1 expression.

Cell Line, Tumor ; DNA Methylation ; Genes, Wilms Tumor ; Humans ; Leukemia ; genetics ; Polymerase Chain Reaction ; Promoter Regions, Genetic