Objective To explore the effect of sodium butyrate(NaB) on phosphorylation/ acetylation of histone H3(ph/acH3) at G?-globin gene and A?-globin gene promoter regions in K562 cells.Methods K562 cells were devided into 2 groups:K562 cells were grown in the presence or absence of 0.5 mmol?L-1NaB for 48 h [K562(NaB) group] and untreated K562 cells group(K562 group).Semi-quantitative RT-PCR was employed to measure the levels of G?-globin mRNA and A?-globin mRNA.The real time PCR-based chromatin immunoprecipitation(ChIP) was used to detect the levels of ph/acH3 at G?-globin gene and A?-globin gene promoter regions.Results Compared with the K562 group,there was a 1.4-fold(t=-149.022,P=0.000) and 1.2-fold(t=-13.363,P=0.000) increase in G?-globin mRNA and A?-globin mRNA,respectively,in K562(NaB) group.The level of ph/acH3 at G?-globin gene and A?-globin gene promoter region increased by 2.9-fold(t=-12.833,P=0.006) and 3.2-fold(t=-10.484,P=0.000),respectively,in K562(NaB) group,compared with the K562 group.The %Input value of G?-globin and A?-globin promoter fragment was 10.0-fold(P=0.000) and 9.5-fold(P=0.000) higher than that value of Necdin gene promoter fragment in the K562(NaB) group,while the %Input value of G?-globin and A?-globin promoter fragment was 3.2-fold(P=0.000) and 2.7-fold(P=0.000) higher than that value of necdin gene promoter fragment in K562 group.Conclusions NaB improves the phosphorylation and acetylation of H3 at ?-globin gene promoter regions,and this may be one of the mechanisms of expression of ?-globin genes induced by NaB.

Cri-du-Chat Syndrome ; complications ; Humans ; Infant, Newborn ; Male ; XYY Karyotype

AIM To investigate the effect of allicin on the regulation of VEGF mRNA expression in human hepatocellular carcinoma cells. METHODS Hepatocellular carcinoma cells were treated with the concentration 10 ?g?L -1 allicin in culture medium,and then the relative VEGF mRNA level at 8 h in human hepatocellular carcinoma cells was evaluated by reverse transcriptase polymerase chain reaction using HPRT(hypoxanthine phosphoribosyltransferase)as an internal control standard. RESULTS The expression of VEGF gene mRNA was inhibited obviously by allicin. Compared with control group, the relative expression level of VEGF gene mRNA was decreased by about 66 36%( P

1)which represented 340 genes and 171 down-regulated(SLR

Objective To explore the effects of astragalus polysaccharides(APS) on fetal hemoglobin(HbF) synthesis and cell proli-feration in K562 cells.Methods K562 cells were chosen as the cell model and cells treated with Na-butyrate(NaB) were taken as the po-sitive control.Western blot was applied to study the level of HbF expression in K562 cells and Trypan blue dye exclusion test was employed to analyze the influence of APS(150 mg/L,300 mg/L,450 mg/L)on K562 cells proliferation.Results 1.Dosage effect:when compared with untreated K562 cells,the HbF expression level increased to(1.56?0.03),(1.78?0.04) and(1.51?0.32) fold,respectively after 48 h treated with different concentrations of APS(150 mg/L,300 mg/L,450 mg/L,F=310.476 P=0).The best inducing concentration was 300 mg/L(P=0.005).2.Time course: HbF levels raised up gradually and the maximum was(2.88?0.27) fold over baseline(P=0) at 48-60 h in the presence of 300 mg/L APS.Then it went to decline.There was statistical significance of HbF expression between K562 cells treated with 300 mg/L APS or NaB [(2.88?0.27) folds,P=0].3.Effects of APS on K562 cells proliferation:the highest reduction of the cell proliferative was obtained in K562 cells cultured in the presence of 0.5 mmol/L NaB.As detected by Trypan blue exclusion met-hod,growth rate of cells stimulated by APS was affect in a dose dependent manner,and significantly higher than NaB.For example,the inhibition rate at 48 hours was 20.45% for 300 mg/L APS but 79.55% for 0.5 mmol/L NaB(P=0).Conclusion APS has ability to induce HbF synthesis in K562 cells and revealed less cells reduction than that of NaB.

Objective To investigate the role of directly constitutive activation of p38 mitogen-activated protein kinases(p38MAPKs)signaling in ?-globin gene expression and fetal hemoglobin(HbF)induction,and provide direct data for the relationship between phosphorylation of p38 and erythroid differentiation of human K562 erythroleukemia cells.Methods The human K562 erythroleukemia cells were transfected with pCDNA 3.1-MKK3(Glu)and pCDNA 3.1-MKK3(Ala)recombinant plasmids by lipofectamineTM 2000.Then,the stable cell lines overexpressing constitutively active p38 and constitutively inhibitive p38 activation were established by the addition of G418 to select single cell G418-resistant clones and identification with reverse transcriptase-polymerase chain reaction(RT-RCR)and Western blot assays,named K562-MKK3(Glu)and K562-MKK3(Ala)cells,respectively.Furthermore,the direct effects of constitutively active p38 on the ?-globin gene expression and HbF induction were analyzed by RT-PCR and benzidine staining,respectively.Results The results of RT-PCR and Western blot showed that there were no evident changes in the mRNA and protein levels of p38 for various cell models,but compared with K562,K562-vect,and K562-MKK3(Ala)cells,the phosphorylation of p38 and expression of ?-globin levels in K562-MKK3(Glu)cells were significantly up-regulated.The results of benzidine staining displayed that the mean percentages of positive cells stained by benzidine in K562,K562-vect,K562-MKK3(Ala),K562-MKK3(Glu)cells,and K562-MKK3(Glu)cells treated with SB203580 were(3.2?1.4)%,(3.7?1.2)%,(2.8?0.9)%,(32.6?5.3)%,and(7.8? 2.3)%(q = 7.56 P

OBJECTIVETo develop a real-time PCR-based chromatin immunoprecipitation (ChIP) assay for determining the effect of sodium butyrate on acetylation of histone in gamma-globin gene promoter regions in K562 cells.

METHODSK562 cells were grown in the presence or absence of 0.5 mmol/L sodium butyrate for 48 h, and 1=10(7) cells per group were used for real-time PCR-based ChIP with anti-acetylated histone H3 or H4 antibodies. The levels of acetylated histone H3 and H4 (acH3 and acH4) in Ggamma- and Agamma-globin gene promoter regions were measured.

RESULTSIn the K562 cells with sodium butyrate treatment or without any treatment, the levels of acH3 or acH4 in Ggamma- or Agamma-globin gene promoter were higher than that in the necdin gene (negative control). Compared with the untreated K562 cells, the cells treated with 0.5 mmol/L sodium butyrate showed a 3.1-fold or 2.6-fold increase in acH3 or acH4 in Ggamma-globin gene promoter region, with also a 3.7-fold or 3.2-fold increase in acH3 or acH4 in Agamma-globin gene promoter region, respectively (P<0.01).

CONCLUSIONWe have successfully developed a real-time PCR-based ChIP assay for analyzing the acetylation of histone H3 and H4 in gamma-globin gene promoter regions. Our results support the role of sodium butyrate in increasing the level of acetylated histone in gamma-globin gene promoter regions.

Acetylation ; Butyrates ; pharmacology ; Chromatin Immunoprecipitation ; methods ; Histones ; chemistry ; Humans ; K562 Cells ; Promoter Regions, Genetic ; genetics ; Real-Time Polymerase Chain Reaction ; methods ; gamma-Globins ; genetics

Objective To investigate the effect of once yearly zoledronic acid of 5 mg on postmenopausal women with osteoporosis of different causes. MethodsFrom October 2009 to December 2009,a total of 89 postmenopausal women with osteoporosis were enrolled and assigned into 2 groups.There were 45 cases of primary postmenopausal osteoporosis,including 27 cases of fresh fracture,in group A.They were aged from 47 to 83 years,with an average of 63.7 years.There were 44 cases of secondary postmenopausal osteoporosis,including 28 cases of fresh fracture,in group B.All patients were given a.single 30-minute intravenous injection of zoledronic acid (5 mg),supplemented by 1,25-dihydroxyvitamin D of 0.25 μg and calcium of 600 mg with VitD125 IU daily.At pre-intervention and 12 months after intervention respectively,bone mineral density (BMD) was measured by dual-X-ray absorptiometry (DXA) at the lumbar spine and hip,and a balance test(Sunlight Tetrax- Ⅱ) was performed to evaluate the risk of falling.Intervention compliance of the patients and adverse events related to zoledronic acid infusion were observed. Results All cases of fresh fracture healed well at 3-month follow-up.At 12 months,43 subjects in group A and 42 subjects in group B completed the follow-up.In group A,BMD increased by 5.8％ at the lumbar spine,by 2.9％ at the femoral neck,by 5.2％ at the Words area,by 5.3％ at the greater trochanter and by 3.9％ at the total hip while the risk of falling decreased by 26.1％; in group B,BMD increased by by 3.4％ at the lumbar spine,by 2.1％ at the femoral neck,by 3.2％ at the Words area,by 3.0％ at the greater trochanter and by 2.5％ at the total hip while the risk of falling decreased by 21.8％.The differences between pre-intervention and post-intervention were significant in both groups ( P ＜ 0.05).No intolerable adverse events occurred in both groups except that one new fracture happened in each group but responded to conservative treatment.ConclusionA once-yearly infusion of zoledronic acid of 5 mg is a convenient and effective therapy for treatment of osteoporosis in postmenopausal women.

Objective\ To investigate apoptosis of Hepatoma BEL\|7402 cells induced by allicin. Methods\ The inhibit effect of allicin on BEL\|7402 cells was investigated with MTT assay. Apoptosis was ascertained by cell morphology under light microscope, electron microscope and flow cytometry. Results\ BEL\|7402 cells were suppressed with dose and time dependent relationship after exposed to allicin of 10,20,40 mg/L for vairous lengths of time. Exposed to allicin at 60 mg/L for 3 hours, BEL\|7402 cells showed typical morphology characters of apoptosis: the microvilli of the cell surface disappeared, the cell shrink and decreased in volume and blebs or ball\|like bodies appeared. Typical nuclear condensation, margination and fragmentation were observed. Apoptotic cells were of trypan blue\|rejected staining. Percentage of apoptotic cells of control group and inductive group was 2\^02?0\^37%,78\^48?3\^15% respectively by trypan blue, and 1\^78?0\^48%, 74\^07?3\^94% by flow cytometry. Cells in G\-0/G\-1, S, G\-2/M phases of control group before induction were 47\^66?2\^72%, 22\^06?2\^04%, 30\^25?3\^78% respectively.Each of them is lower than the percentage of apoptotic cells. Conclusion\ Allicin induced apoptosis of hepatoma BEL\|7402 cells.Apoptotic cells were supposed to be initiated in some phases of cell cycle.\;[