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

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

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.

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

ObjectiveTo investigate the effects of caleitriol,training on balance and lower extremity muscle strength on fall risk of postmenopausal women with osleoporosis or osteopenia.Methods A total of 200 postmenopausal women with osteoporosis or osteopenia,whose balance test confirmed higher fall risk,were randondy assigned to group A or B.Those of group A received the following intervention:( 1 ) 0.25 μg calcitriol,QD; (2) general information on fall and osteoporosis; (3) balance training; (4) lower extremity muscle strength exercises.Those of group B were only treated with 0.25 μg calcitriol.All the participants were supplemented with 600 mg/d calcium and 125 IU/d vitamin D.Fall index,bone mineral density,serum levels of calcium and phosphorus,and adverse reactions were record.Results After 3 months' intervention,the fall index of both groups was significantly decreased ( group A:t =2.16,P＜0.05 ; group B:t =2.08,P＜0.05 ).After 6 months' intervention,the fall index of both groups went on decreasing,and significant difference of fall index between 6 month and baseline of group A and between group A and group B at 6 months was found.After 1-year intervention,the fall index of group A was further decreased in comparison with group B ( t =2.66,P＜0.05 ).No hypercalcemia occurred during the study period.Conclusion The fall risk of the patients with postmenopausal osteoporosis or osteopenia was reduced after 3 months' intervention.Twelve months' active vitamin D intervention could either reduce the risk of fall or improve bone mineral density.Patient education,balance training and muscle exercise may be effective intervention to reduce fall risk.

Objective To study on the effectiveness of endoscopic ultrasonography(EUS)in detec- ting insulinoma preoperatively.Methods Fifteen patients with clinical and biochemical signs of insulinoma were examined by EUS using a radial-scanning ultrasound endoscope and abdominal ultrasonography,CT, DSA prior to surgery.The outcome was evaluated on the basis of surgery and examination of the resected specimens.Results Fifteen patients with 16 lesions of insulinoma were identified by surgery and pathology. The aceuraey of diagnosis with EUS was 13/15(86.7%),and that with B-US,CT,DSA was 3/15(20%), 5/15(33.3%),9/14(64.3%)respectively.In the 14 lesions identified by EUS,10 lesions were depicted to be hypoechogenic,1 lesion was isoechogenic and 3 lesions were hyperechogenie.All 14 lesions were well demarcated and surrounded by normal pancreatic tissue.The minimum size of the lesion visualized by EUS was 0.5cm.Ten lesions were correctly detected by EUS with size of 0.5～2.0cm.EUS missed diagnosis in 2 lesions not for their small size.EUS falsely indicated a 10mm lesion from two lesions inside the head of pancreas.One lesion outside the pancreatic tail and one lesion in the pancreatic head were missed by EUS in another case.Conclusion EUS is superior in assessing the location of pancreatic insulinoma than other ima- ging methods such as B-US,CT,DSA.