Objective To analyze the role of hDOT1L signaling pathway on the proliferation of human acute monocytic leukemia cell line THP-1 cell inhibited by demcitabine, and to explore the other effects except DNA demethylation and the mechanism of hDOT1L signaling pathways involved in leukemia. Methods Methyl thiazolyl tetrazolium (MTT) method was used to detect the influence of decitabine on THP-1 cell proliferation, and trypan blue anti-staining was applied to detect the effect of decitabine on THP-1 cell survival rate, reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect the expression levels of hDOT1L, HOXA9, HOXA10, MLL-AF10 (MLLT10) mRNA, then the methylation level of H3K79 was detected by Western blotting assay. Results Compared with the control group, decitabine inhibited the proliferation of THP-1 cell in a time and dose-dependent manner. The inhibitory effect of 72 h was the highest, and the highest inhibition rate was 56.18%. In addition to 0.5 μmol/L in decitabine treatment group, the other groups were statistically significant (all P<0.05). There were high expressions of hDOT1L, HOXA9, HOXA10, MLLT10 mRNA and H3K79 hypermethylation in the THP-1 cell, meanwhile decitabine reduced the levels of hDOT1L mRNA after 48 h as well as HOXA9, HOXA10, MLLT10 mRNA, and there was a significant difference compared with the control group (all P < 0.01). Decitabine reduced obviously the methylation of H3K79 in cell after 48 h, especially the expression levels of bis and three methyl protein, and these differences were significant compared with the control group (all P< 0.01). Conclusion Decitabine plays its inhibitory effect on the proliferation of THP-1 cells probably through reducing the expression level of hDOT1L and H3K79 methylation level and decreasing the expression levels of key molecules, HOXA9, MLL-AF10, HOXA10, MLLT10 in the hDOT1L signal pathway.

Background Erythropoietin (EPO) was proved to be express in hematopoietic tissue and nervous system and play the effects of stimulating blood cell production and protecting nervous tissue.Researches showed that EPO is expressed in the embryon brain of animal.However,whether EPO exist in nervous-derived retina and its action on retina with the development is concerned. Objective This research was to investigate the expression of EPO during the embryonic development period of rat retina and explore the role of EPO in retina development process.Methods Clean Wistar rats with pregnancy for 12 days,16 days and 20 days were collected,and the embryonic 12-day rats (E12 d,5 rats),embryonic 16-day rats (E16 d,5 rats) and embryonic 20-day rats ( E20 d,5 rats) were obtained by caesarean operation,and 5 12-month W istar rats were used as controls.The rats were sacrificed by cervical dislocation and the retinal sections were prepared in the different-embryo-phase (12 d,16 d,20d) and growth phase.The expression of EPO protein and mRNA in rat retina was detected by immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR),respectively.The feed and use of the animals followed the Regulation for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results EPO was positively expressed in the cytoplasm and nuclei in the neuroepithelial layer and pigment epithelium of every-embryo-phase rats but only in retinal ganglion cell layer in 12-month-old rats.The gray scale values of EPO expression in retina were 105.55±10.35,99.35± 8.71,83.27± 7.84and 30.30± 3.80 in E12 d rats,E16 d rats,E20 d rats and 12-month-old rats respectively with a statistically significant difference (F=76.13,P＜0.01 ).RT-PCR revealed that the relative values of EPO mRNA expression in retina were 0.876±0.10,0.861 ±0.09 and 0.256±0.03 in E16 d rats,E20 d rats and 12-month-old rats respectively,presenting a elevated value in embryonic rats compared with adult rats ( P =0.00).Gel imaging deletion showed that the A value of EPO amplification products was highest in E16 d rats and lowest in adult rats.Conclusions The expression of EPO appears a high to low fashion during the embryonic development of Wistar rats,which is closely associated with the developing procedure of retina.

Objective To detect the genetic amplification and protein expression characteristics of aurora-A in ovarian cancer and to interpret the role of aurora-A gene in course of onset,progression and regression phases of ovarian cancer.Methods The amplification of aurora-A gene was detected by quantitative PCR in 6 normal ovarian tissues and 8 ovarian cancer samples,and its protein expression was examined by immunohistochemistry in 6 normal ovarian tissues and 40 ovarian cancer samples,furthermore,the relationships between over-expression of aurora-A protein in ovarian cancer tissue and its pathologic classification,tissue differentiation,clinical phase,tumor proliferation trait and prognosis were analyzed.Results Quantitive PCR showed that aurora-A mRNA was significantly higher in 8 ovarian cancer samples than that in normal ovarian tissues(P0.05).Conclusion There are abnormal amplification and protein over-expression of aurora-A in ovarian cancer tissue,aurora-A probably play an important role in the onset and progression of ovarian cancer,and the novel biological treatment concerning aurora-A gene and its protein is probably a useful route for curing tumor.

BACKGROUND: Basic fibroblast growth factor (bFGF), a kind of polypeptide growth factor possessing multifunctional biological activities,can protect neurons and promote the growth of nerves. It has been corfirmed that bFGF has therapeutic effects on retina ischemia/reperfusion injury (RIRI).OBJECTIVE: To establish RIRI model and analyze the effects of bFGF on cellular apoptosis of retina and the expression of regulatory gene protein.DESIGN: Randomized grouping and validating trial.SETTING: Department of Ophthalmology, the Affiliated Hospital of Medical College of Qingdao University.MATERIALS: The experiment was conducted at the Research Laboratory of Pathology, Department of Ophthalmology, Medical College of Qingdao University, from April 2002 to December 2003. Twenty-eight healthy Wistar rats were enrolled in this experiment. Four rats were randomly chosen for normal control group, the left eyes of the other 24 rats were set as normal saline control group, and the right eyes were set as bFGF group.METHODS: Normal saline control group and bFGF group adopted the rat RIRI models established by transiently elevating intraocular pressure. Normal saline of 12 μL was injected into the vitreous cavity of the left eyes of the rats in normal control group. 12 μL bFGF was injected into the vitreous cavity of the right eyes of the rats in bFGF group, 4 rats once. No administration was given in normal control group. The expression of apoptotic cells was detected and apoptosis indexes were calculated with the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling (TUNEL) method and immunohistochemical staining method at the 1st, 6th,12th, 24th,48th and 72nd hours after reperfusion and ischemia for 1 hour.MAIN OUTCOME MEASURES: ① The detection results of apoptotic cells in situ of retina tissuesat different time points after reperfusion. ②The expression of Fas and caspases-2 in retina tissues at different time points after reperfusion.RESULTS ① Comparison of apoptosis indexes of retina tissues at different time points after ischemia reperfusion: There were no apoptotic cells in the retina tissues of the rats in normal control group. As compared with those in normal saline control group, apoptosis indexes in bFGF group were significantly decreased at ischemia 1 hour and reperfusion 1, 6, 12, 24, 48and 72 hours, especially at the 12th, 24th and 48th hours after reperfusion (t =5.362-5.595, P ＜ 0.05). ② The change of Fas expression at different time points after ischemia reperfusion: There was hardly any Fas expression in normal control group. As compared with that in normal saline control group, Fas expression in bFGF group was significantlydecreased at ischemia 1 hour and reperfusion 1, 6, 12, 24, 48 and 72 hours, especially at the 6th, 12th and 24th hours after reperfusion (t=3.954-9.327, P ＜ 0.05). ③The changes of caspase-2 expression at different time points after ischemia reperfusion: There was no caspase-2 expression in normal control group.Compared with that in normal saline control group, the number of caspase2 positive cells in bFGF group was significantly decreased at the 6th,12th,24th, 48th and 72nd hours after ischemia for 1 hour and reperfusion (t=4.125-15.641, P ＜ 0.05).CONCLUSION: bFGF can significantly inhibit the expression of apoptosis gene Fas and caspase-2 in the ischemia and reperfusion of retina, thus reducing cellular apoptosis of ganglion cells and exerting therapeutic effects on the ischemia and reperfusion of retina.

Adult ; Ethylene Oxide ; poisoning ; Female ; Humans ; Occupational Exposure

AIM: To explore the relationship between the expression of caspase-2 and caspase-3 and the apoptosis in retinal ischemia/reperfusion (I/R) injury of rats, as well as the therapeutic effects of brain derived neurotrophic factor (BDNF)on the ischemic and reperfused retina.METHODS: This experiment was conducted at the laboratory of Affiliated Hospital of Qingdao University Medical College from February 2007 to July 2007. The models of retinal ischemia/reperfusion injury were made by transiently elevating intraocular pressure. A total of 28 rats were divided into Normal and Operative Groups. Operative group was divided into six subgroups. In each subgroup there were four rats. The left eyes of rats were used for I/R and the right eyes were used for intravitreal injection of brain-derived neurotrophic factor (BDNF) as treatment group. After reperfusion we divided our subgroups according to the reperfusion time as 1, 6, 12, 24, 48, 72 hours. The retinal ganglion cell number was counted by using optic microscope(BX-51,Olympus). Apoptosis was assessed by the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labelling (TUNEL) method, and the expression of caspase-2,caspase-3 was studied by enzyme linked immunosorbent assay (ELISA) and strept avidin-biotin complex (SABC)immunohistochemistry.RESULTS: No positive apoptotic cells were observed in the normal rats' retinae, but there were a significant number of positive apoptosis cells in 6-24 hours after transient ischemia followed by a decrease at 48 hours. The number of apoptotic cells reached a maximum at 24 hours after ischemia .The expression of caspase-2 gradually increased as early as at 6 hours, reached a peak at 24 hours, then decreased between 48 and 72 hours. Similarly, caspase-3 has the same rule with caspsae-2 in the time courses of expression in retinal tissues.BDNF administered before reperfusion inhibited the expression of apoptosis and ameliorated the retinal tissue damage. It also decreased caspase-2 and caspase-3 expression in ischemic/reperfused retina.CONCLUSION: Retinal ischemia-reperfusion can induce apoptosis of cells in the retina. BDNF rescues retinal ganglion cells (RGCs) from retinal ischemia/reperfusion injury through down-regulation of cell apoptosis and caspase-2 and caspase-3 expression. BDNF have a neuroprotective effect on retina.

Objective Previous studies indicated that activation of Toll-like receptor4 (TLR4) was involved in the progression and instability of atherosclerotic plaque.Anti-inflammatory effects were shown in statins. However,the mechanisms underlying these effects have not been well explored.We test the hypothesis that a por- tion of these anti-inflammatory effects are mediated by regulation of TLR4 expression.Methods One hundred twenty-one subjects (22 normal persons,17 patients with stable angina and 82 patients with ACS) were recruited. 41 patients with ACS were randomized to atorvastatin 10 mg/d or atorvastatin 40 mg/d on top of routine anti-anginal treatment.Serum level of hsCRP,blood lipids,TLR4 expression on CD_(14)~+ monocytes were measuered before and after one month treatment.TLR4 expression on CD_(14)~+ monocytes were quantified via flow-cytometry.Results hsCRP and TLR4 expression on CD_(14)~+ monocytes in patients with ACS were higher than patients with stable angina and normal persons(hsCRP,ACS:11.1?14.3 vs stable angina:2.5?2.7 mg/L vs normal:2.3?4.2 mg/L,P

Objective To investigate the expression and significance of microRNA - 21(miR - 21)in acute kidney injury mice model at the different time points following ischemic/ reperfusion. Methods C57BL/ 6J mice were divided into 3 major groups:the control group(C group),sham operation group(S group)and ischemia - reperfusion group(IR group). Later 2 groups were divided into 9 sub - groups respectively according to the time following reperfu-sion. Automatic biochemical analyzer detected serum creatinine(Scr),blood urea nitrogen(BUN)level. HE staining detected renal pathological damage. Renal tubulointerstitial pathological score accessed pathological damage. Real time - PCR tested the expression of miR - 21 and mitogen - activated protein kinase kinase 3(MKK3)mRNA in renal respectively. Immunohistochemistry staining tested expression of MKK3. Results IR group's Scr,BUN levels gradually increased following reperfusion,24 h reached its peak,then gradually declined. The Scr,BUN level had statistically sig-nificant difference between IR group and S group at the same time subgroup from 3 h to 168 h following reperfusion(all P ﹤ 0. 01). The change of kidney damage and pathological changes of interstitial and tubular injury score consensus with renal function. miR - 21 increased gradually in renal ischemia after reperfusion,24 h peaked and then stabilized at this high level. miR - 21 was positively correlated with pathological tubulointerstitial injury from 0 h to 168 h after reperfu-sion(r = 0. 969,P ﹤ 0. 05). IR group's MKK3 mRNA and protein expression rose sharply following ischemia/ reperfu-sion,24 h peaked,and then gradually decreased. From 3 h to 168 h,the expression of MKK3 mRNA and proteins had significant difference at each same time points subgroups between IR group and S group(all P ﹤ 0. 01). Conclusions miR - 21 increases gradually in renal ischemia after reperfusion,24 h peaked and then stabilized at this high level. miR - 21 is positively correlated with pathological tubulointerstitial injury,which may be associated with the negative regulated relationship between miR - 21 and MKK3.

Objective To investigate the correlation between retinal deformation degree and retina thickness (RT) detected by optic coherent tomography (OTC) in patients with idiopathic macular epiretinal membrane (IMEM). Methods The 66 eyes (56 patients) with IMEM diagnosed by OCT were retrospectively analyzed in this study. After best corrected visual acuity (BCVA), ocular fundus and OCT examination, patients were divided into three groups (mild, medium and severe) according to retinal deformation degrees. The RT value was measured manually. Results There were significant differences in RT values between mild, moderate and severe groups:(311.95 ±51.78) μm, (447.13±41.95) μm and (560.00± 58.23) μm (P<0.05). The values of BCVA were 0.78±0.16, 0.38±0.12 and 0.27±0.14 for mild, moderate and severe groups, there were significant differences between them (P<0.05). There was no significant correlation between RT and BCVA in mild group (r=-0.352,P=0.128). There was negative correlation between RT and BCVA in medium group and serious group (r=-0.768 and-0.482,P<0.05). Conclusion The retinal deformation degree and RT are two objective indicators to assess visual performance in patients of IMEM. When RT is more than 400 μm, it can be used as objective criteria for surgical intervention.

OBJECTIVETo investigate the effect of insulin-like growth factor-I (IGF-I) on the proliferation and osteogenesis of human periodontal ligament stem cells (hPDLC) under three-dimensional (3D) culture system.

METHODSHuman periodontal cells were isolated from the ligament of surgically extracted human teeth, and through the limiting dilution assay, got mono-clone of the cell, hPDLCs were isolated from MesenPRO RS medium. Rotary cell culture system (RCCS) was enrolled to set 3D environment. Control group and experiment groups were assigned according to the concentration of IGF-I. There were 5 level of experiment groups (0.1, 1, 10, 50, 100 µg/L). Proliferation was tested with methyl thiazolyl tetrazolium (MTT), and alkine phosphatase (ALP) level was assayed by spectrophotometer to analyze the osteogenesis of hPDLCs. Gene expression of ostetocalcin (OCN) and type I collagen (Col I) were assayed by reverse transcriptase polymerase chain reaction (RT-PCR).

RESULTSIn 3D culture system, the effect of IGF-I on cell proliferation was significantly different between control group and experiment groups (P < 0.05), and there showed significant differences between the group of 0.1 µg/L (0.219 ± 0.021) IGF-I and the groups of 50, 100 µg/L (0.287 ± 0.011, 0.293 ± 0.012). However, there showed no significant differences among other groups. Significant differences of ALP activity were observed between the control group and experiment groups, and between the groups of 1, 10 µg/L (0.304 ± 0.020, 0.310 ± 0.013) and that of 50, 100 µg/L (0.347 ± 0.011, 0.344 ± 0.010) (P < 0.05). While no significant differences were detected between the group of 1 µg/L and that of 10 µg/L, nor between the group of 50 µg/L and that of 100 µg/L. Expressions of Col I and OCN in mRNA and protein level both showed dose-dependent increase.

CONCLUSIONSIn 3D culture system, in the scale of 0.1 - 100 µg/L, the effect of IGF-I on the proliferation of hPDLCs increased dose-dependently. 100 µg/L IGF-I promotes osteogenesis of the cells significantly.

Alkaline Phosphatase ; metabolism ; Cell Culture Techniques ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen Type I ; genetics ; metabolism ; Dose-Response Relationship, Drug ; Humans ; Insulin-Like Growth Factor I ; pharmacology ; Osteocalcin ; genetics ; metabolism ; Osteogenesis ; drug effects ; Periodontal Ligament ; cytology ; RNA, Messenger ; metabolism ; Stem Cells ; cytology ; metabolism