Objective To explore the relationship between cytogenetics with therapy and prognosis in myelodysplastic syndrome.Methods 124 cases of myelodysplastic syndrome from September 2005 to October 2009 were reviewed.124 cases contained 79 males and 45 females,aged 14 to 79 years,median age was 57 years old.26 cases were diagnosed as RA,13 cases as RAS,21 cases as RCMD,29 cases as RAEB-Ⅰ,35 cases as RAEB-Ⅱ.According to the karyotype,124 cases were divided into two groups,79 cases of normal karyotype and 45 cases of abnormal karyotype.Patients were followed up for three years in order to analyze the incidence of myelodysplastic syndrome changed into acute leukemia.Results In 39 patients diagnosed as RA and RAS,32 cases were with normal karyotype and 7 cases (17.9 ％) with abnormal karyotype.15 of 32 cases with normal karyotype achieved hematologic remission after treatment while only 1 of 7 cases with abnormal karyotype achieved remission.1 case with abnormal karyotype changed into acute leukemia after 2 years.In 85 patients diagnosed as RCMD,RAEB-Ⅰ and RAEB-Ⅱ.Difference of the lower incidences of RA and RAS changed into acute leukaemia during 3 years in normal karyotype and abnormal karyotype groups was statistically insignificant (P ＜ 0.05).The incidences of RCMD,RAEB-Ⅰ and RAEB-Ⅱ changed into acute leukemia were higher,especially in the abnormal karyotype group with 42.1 ％.Conclusion Myelodysplastic syndromea with abnormal karyotype is associated with poorer efficacy of therapy,higher incidence of changing into acute leukaemia.The patients can take early allogeneic hematopoietic stem cell transplantation to improve the prognosis.

Objective To explore ways for the supervision of department directors. Methods Job responsibility agreements with department directors were signed and a series of assessment standards were established. Results The methods adopted were standardized, individualized and easy to use. Conclusion Job responsibility agreements with department directors and a standard assessment system are currently effective measures for the supervision of department directors.

OBJECTIVE:To establish an analytical method for the determination of prednisolone and prednisolone-hemisuccinate in biological media METHODS:Using RP-HPLC,biological media were extracted with a mixture of n-pentane and methyl tert-butyl ether(2∶3) The mobile phase consisted of acetonitrile and trisodium citrate buffer (33∶67) The sample was eluted on C18 column and detected at 254nm RESULTS:The extraction recoveries of two drugs were over 80% and the RSDs of within-day and day-to-day were below 3% CONCLUSION:The method is simple,rapid,sensitive,accurate and good enough to be applied to the pharmacokinetic study of new preparation of prednisolone and can also be applied to other similar glucocorticoids

AIM: To explore the expression changes of bone morphogenetic protein-7 (BMP-7) and its receptors (BMPR-Ⅱ, ALK-2, ALK-3, ALK-6) in the renal tubulo-interstitial lesions induced by unilateral ureteral obstruction (UUO). METHODS: Rats were divided into normal control, sham operation and UUO groups, and sacrificed at postoperative day 1, 3, 7 and 14. The mRNA levels of BMP-7, BMPR-Ⅱ, ALK-2, ALK-3 and ALK-6 were examined by RT-PCR. The protein expression site and level of BMP-7 was detected by immunohistochemistry staining. RESULTS: Compared to sham operation group, the mRNA levels of BMP-7, BMPR-Ⅱ, ALK-2 and ALK-3 were significantly decreased, but the change of ALK-6 mRNA was not marked in UUO rats. The reduction of mRNA expression levels of BMP-7, BMPR-Ⅱ, ALK-2 and ALK-3 in the kidney tissue aggravated along with the delayed post-operated days. The results of immunohistochemistry staining indicated that BMP-7 mainly expressed in renal tubular and interstitial, rarely in glomeruli. In UUO rats, the protein expression of BMP-7 decreased in varying degrees according to the post-operated days. CONCLUSION: The loss of BMP-7 and its receptors (BMPR-Ⅱ, ALK-2, ALK-3) were observed in the early phase of fibrotic process and this may play a very important role in mediating the renal tubulointerstital fibrosis.

Objective To explore the role of bone morphorgenetic protein-7(BMP-7) in the renal tubulointerstitial lesions induced by unilateral ureteral obstruction (UUO).Methods After dividing into normal control, sham operation and unilateral ureteral obstruction (UUO) groups, sixty rats were sacrificed at postoperative day 1, 3, 7, 14. The levels of BMP-7 and TGF-?1 mRNA were examined by RT-PCR. The sites and levels of protein expression of BMP-7, TGF-?1 and ?-SMA were detected by immunohistochemistry staining. Results Compared to control groups, BMP-7 mRNA was significantly decreased, but TGF-?1 mRNA was significantly increased in UUO rats. Immunohistochemistry staining studies indicated that BMP-7 mainly expressed in renal tubule and interstitium, rarely in glomeruli. In UUO rats, the protein expression of BMP-7 decreased, but the expression of TGF-?1 and ?-SMA increased in varying degrees according to the obstructed days. The renal tubulointerstitial expression of BMP-7 was significantly negatively correlated with the expression of TGF-?1 and ?-SMA. Conclusion The loss of BMP-7 is explored in the early phase of fibrotic process and may play a very important role in mediating the tubulointerstital lesions.

The transfection efficiency of oligonucleotide and plasmid to the HL-60 cell line with lipofectaminePLUS was compared through observing the transfection rate and the expression duration of exogenous gene in the target cells. The results showed that the transfection rate of oligonucleotide to the HL-60 was about 90% - 95% and it had no obvious attenuation within 84 h. However, the plasmid transfection rate was only 5% -25% and it was decreased significantly within 60 h. It was suggested that the transfection of oligonucleotide with liposomes was better than that of plasmid.
Green Fluorescent Proteins/genetics ; HL-60 Cells ; Liposomes ; Oligonucleotides/*genetics ; Plasmids/*genetics ; Transfection

In order to explore whether gene CHFR was inactivated by methylation in leukemia cells, the expression of CHFR was examined before and after treatment with demethylation agent in Molt-4, Jurkat and U937 leukemia cell lines by means of RT-PCR. The methylation of promoter in Molt-4, Jurkat and U937 cells as well as 41 acute leukemia patients was analyzed by MS-PCR. The results showed that methylation of CHFR promoter was inactivated and could be reversed by treatment with a demethylating agent in Molt-4, Jurkat and U937. CHFR promoter methylation was detected in 39% of acute leukemia patients. There was no difference in incidence of CHFR promoter methylation between acute myelocytic leukemia and acute lymphocytic leukemia. In conclusion, CHFR is frequently inactivated in acute leukemia and is a good candidate for the leukemia supper gene. By affecting mitotic checkpoint function, CHFR inactivation likely plays a key role in tumorigenesis in acute leukemia. Moreover, the methylation of gene CHFR appears to be a good index with which to predict the sensitivity of acute leukemia to microtubule inhibitors.
Cell Cycle Proteins/*genetics ; DNA Methylation ; DNA, Neoplasm ; Epigenesis, Genetic ; Leukemia, Myeloid, Acute/*genetics ; Neoplasm Proteins/*genetics ; Precursor Cell Lymphoblastic Leukemia-Lymphoma/*genetics ; Promoter Regions (Genetics)/*genetics ; Tumor Cells, Cultured

The HL-60 cells were transfected with chk1 antisense and sense chain, and 24 h later subjected to irradiation. Twenty-four h after irradiation, the changes in the chk1 protein expression was assayed by Western blot, and the cell cycles and apoptosis rate detected by FCM. The irradiated apoptosis sensitivity was increased by antisense blocking of chk1 gene in HL-60 cell line with the apoptosis rate being 26.31%, significantly higher than that by the sense blocking (10.34%, 0.025 < P < 0.05). In HL-60 cells transfected with chk1 antisense chain, the G2/M phase arrest was attenuated and the cells in G2/M phase were accounted for 38.42%, significantly lower than those of the cells transfected with chkl sense chain (54.64%, 0.005 < P < 0.01). It was concluded that antisense blocking of chk1 gene could increase the apoptosis sensitivity to irradiation.
*Apoptosis/radiation effects ; Cell Cycle/radiation effects ; HL-60 Cells ; Oligonucleotides, Antisense/*genetics ; Protein Kinases/*genetics ; Protein Kinases/metabolism ; Radiation Tolerance/*genetics ; Transfection

Silencing ATM gene gave rise to enhanced apoptotic response to irradiation and irradiation-like chemotherapy agents, this paper explored the crucial identities of the molecular elements responsible for the enhanced apoptotic response in U937 cells mediated by silencing ATM gene. Two U937 cell mutants named U937-ASPI3K (ATM, negative) and U937-pZeosv2(+) (ATM, wild-type) were used as a cell model system to identify the critical molecule(s) responsible for the varied apoptotic response in the absence or presence of ATM gene. Apoptosis was examined by measuring concentrations of free nucleosome in U937 cells. Western blot was employed to measure nuclear protein abundance of CDC25A, CDC25B, CDC25C, total p34cdc2, p34cdc2, (Thr 161) or p34cdc2 (Thr 14, Tyr 15). RT-PCR was used to estimate CDC25 transcript levels. U937-ASPI3K exhibited an enhanced apoptotic response to lower dosage of irradiation, which could not be blocked by protein synthesis inhibitor. Protein serine-threonine phosphatase inhibitor or cyclin-dependent kinase (CDK) inhibitors, on the other hand, abolished the enhancement indicated that protein phosphorylation/dephosphorylation modification and CDK activity are required for the enhanced apoptotic response in the absence of ATM gene. Upon irradiation, p34cdc2 in U937-pZeosv2(+) was maintained in an inactive state by phosphorylation on threonine 14 (Thr 14) and tyrosine 15 (Tyr 15), which was associated with a dramatic decrease of nuclear CDC25A, CDC25B and CDC25C proteins. In contrast, p34cdc2 in U937-ASPI3K maintained in an active state by dephosphorylation on threonine 14 (Thr 14) and tyrosine 15 (Tyr 15), which was associated with constant nuclear CDC25A, CDC25B and CDC25C protein abundance before and after irradiation. The responsive decrease of nuclear CDC25 proteins occurred at the post-transcription level. Silencing ATM gene blocks the responsive decrease of nuclear CDC25 proteins, which is responsible for failure to inactivate p34cdc2 after irradiation. Active p34cdc2 and CDK2, in turn, acts as the death executors to trigger apoptosis. In summary, aberrantly activated CDK activity is the critical molecular mechanism central to enhanced apoptotic responses in the absence of ATM gene.

Objective To evaluate the impact of 1125 seeds para-tracheal braehytherapy on regional tissue injury in rab-bit models. Methods 42 rabbits were randomized into 7 groups. Group 1 to 6 belong to study groups (in which 1,4,5 and 6 belong to "dose gradient" subgroup, while 2,3 and 4 to "chronologic" subgroup) , while the last group acts as negative con-trol. The activity of seeds in study group were 0.3 mCi in group 1, 0.5 mCi in group 2 to 5, 0.7 mCi in group 5, and 0.9mCi in group 6. False seeds (0 mCi) were used for the negative control. 4 seeds with equal dosage were implanted between trachea and esophagus in each rabbit under general anesthesia. Seeds arrangement was made according to Paris principle. For the tissue injury evaluation, group 2 was sacrificed by the end of first month post-operatively, group 3 at the end of the second month, and group 4 end of the third month. The rest of rabbits were also sacrificed at the end of the third month. Pieces of adjacent e-sophagus and trachea were sampled from each rabbit. Tissue injury features such as inflammation, edema, congestion or fibrosis as evaluated histologically. Results All rabbits were healthy during study period except 5. Histological analysis revealed that trachea samples from all groups had lymphocytas and plasma cells infiltration as signs of chronic inflammation, hut fibrosis was nut clearly visible. There were no differences between study and control groups with respect to inflammation, edema and con-gestion scores. But in groups which received the highest doses of radiation or sacrificed at 60 d showed more eosinophil infiltra-tion and epithelum degeneration, and statistical significance was reached between these groups and control. Esophageal samples had less histological changes compared with trachea. Conclusion Para-tracheal implantation of ~(125)Ⅰ seeds with therapeutic or higher dosage only induce minor and reversible damage to the regional tissue. This implies that ~(125)Ⅰ implants adjacent to trachea or esophagus are clinically safe.