Aim To study the regulation mechanisms of deacetylase inhibitor SAHA in p21WAF1/CIP1 promoter acetylation in breast cancer MCF-7 cells.Methods We used quantitative real-time PCR, Western blot and DNA-ChIP to determine the effects on the regulation of cell cycle with SAHA treatment in MCF-7 cells.By DNA-ChIP, we assessed the acetylation level of p21WAF1/CIP1 promoter.Results SAHA significantly affected the expression of cell cycle-related factors, and induced the mRNA and protein expression of p21WAF1/CIP1.SAHA could adjust the acetylation level of p21WAF1/CIP1 promoter.Conclusion SAHA regulates the cell cycle progression by adjusting the acetylation level of p21WAF1/CIP1 promoter in MCF-7 cells.

After an elaboration with the ideas and steps for developing a cross platform acupoint display and re-trieval system using flash technology, how to rapidly locate and display the acupoints and how to search the recent papers on acupoint therapy were described with the Jing-Well Point as an example.

Aim Toinvestigatethespecificbinding sites that HDAC1 can be recruited to regulate the tran-scriptional activity of p21 WAF1/CIP1 promoter in the breast cancer MCF-7 cells.Methods ThebreastcancerMCF-7 cells in logarithmic growth phase were starved with FBS free medium for 24 hours,and treated with 20 μmol·L-1 SAHA(S group)or 0. 625 nmol·L-1 Leptin(L group)for 24 hours,and the cells that were cultured in the complete RPMI 1640 medium without any treatment were assigned as control group (B group).The DNA-ChIP was followed the manufactur-er′s protocol for the assay.The cell lysis was prepared and incubated with anti-HDAC1 antibody overnight at 4℃.DNA fragments binding anti-HDAC1 antibody were gathered and purified.The relative expression level of DNA fragments from TSS to the upstream of the p21 WAF1/CIP1 promoter region(+2 ~-4000 bp)bind-ing with antibody was detected by Real-time PCR and analyzedby2-ΔΔCTmethod.Results InBgroup, HDAC1 had high affinity with the f1 and f 8 fragmentsof p21 WAF1/CIP1 promoter compared to the other fragemts,and showed the highest affinity with the f8 fragment.In S group,the binding ability of HDAC1 to the f1 ~f10 fragment of p21 WAF1/CIP1 promoter was sig-nificantly lower than that of the control.The binding activity of HDAC1 to f8 fragment was the lowest,while reversing to reach the peak after leptin treatment.Con-clusions HDAC1canberecruitedtop21WAF1/CIP1pro-moter by the cell proliferation signal during the prolifer-ation of breast cancer MCF-7 cells.The DNA fragment from -2800 to -3200 bp in the upstream of p21 WAF1/CIP1 promoter is the target functional region for the binding with HDAC1 .

Aim To clarify the regulation role of ca-thepsin B ( Cat B ) in cell proliferation and apoptosis induced by SAHA in ER-positive breast cancer cell line MCF-7.Methods MTT was used to screen the optimal concentration and treatment time of SAHA . The expression levels of related proteins were deter-mined by ELISA , and the morphological changes were observed through time-lapse live cell imaging acquisi-tion.Cell viability and apoptosis assay in MCF-7 cells were assessed by Muse Cell Analyzer with SAHA and /or Cystatin C treatment .Results MTT assay showed that the anti-tumor efficacy of SAHA was significant . The optimal concentration and treatment time were 10μmol? L-1 and 24 h respectively . ELISA assay showed that SAHA could induce expression of Cat B in MCF-7 cells.Real-time live-cell imaging experiments demonstrated that the combination treatment of Cystatin C and SAHA significantly resumed the inhibitory effect caused by SAHA alone .Cytology test showed that SA-HA alone obviously depressed the cell viability and in-duced apoptosis . However , the effect was reversed with the combination of Cystatin C .Conclusion Cat B plays an important role in apoptosis induced by SAHA in ER +breast cancer cells MCF-7.

Objective:To observe the clinical efficacy and side effects of highly agglutinative staphylococcin injection at high dose combined with oxaliplatin or cisplatin with intraperitoneal perfusion in the treatment of malignant peritoneal effusion. Methods:Totally 60 patients with malignant peritoneal effusion were divided into two groups,32 cases in group A,and 28 cases in group B. Group A was treated with staphylococcin at high dose combined with oxaliplatin with intraperitoneal infusion after peritoneal cavity catheter drainage,and group B was treated with staphylococcin at high dose combined with cisplatin with intraperitoneal infusion after peritoneal cavity catheter drainage. All patients underwent the treatment once a week,and continuous four weeks for a courese. The clinical efficacy and adverse drug reactions were compared between the two groups. Results:The complete response rate of group A was 18. 8% ,and the total effective rate was 90. 7% ,which were both significantly higher than those of group B(P < 0. 05). The incidence of abdominal pain in group A was higher than that in group B(P < 0. 05),while there were no significant differences in the total incidence of adverse drug reactions and the other symptoms between the two groups(P > 0. 05). Conclusion:Staphylococcin at high dose combined with oxaliplatin with intraperitoneal perfusion is more effective than staphylococcin at high dose combined with cisplatin. Although abdominal pain in group A is obvious,the patients are able to tolerate with it after the symptomatic treatment,therefore,staphylococcin at high dose combined with oxaliplatin with intraperitoneal perfusion can be used as one of effective methods for the treatment of malignant peritoneal effusion.

Objective To investigate the diagnosis of diffusion-weighted imaging (DWI) in cervical lymph nodes lesions.Methods Twenty patients with metastatic lymph node,10 patients with tuberculosis lymph node and 10 volunteers were underwent both conventional MRI and DWI,and the ADC values of lymph nodes' solid components were measured.The difference of ADC values between three kinds of lymph nodes were analyzed.Results ADC values of metastatic lymph nodes was (0.842 ± 0.156) × 10-3 mm2/s,ADC values of tuberculosis lymph nodes was (1.249 ± 0.142) × 10-3 mm2/s,and ADC values of normal lymph nodes was (1.291 ± 0.176) × 10-3 mm2/s.There was significant difference between ADC values of metastatic lymph nodes and with the two later kinds of lymph nodes (P＜ 0.01),and no difference in the two later kinds of lymph nodes (P ＞ 0.05).Conclusion ADC values with features of conventional MR is helpful for differentiate metastatic lymph node,tuberculosis lymph node and normal lymph nodes.

Aim To investigate the specific binding sites that histone deacetylases 1(HDAC1) and estrogen receptor α(ERα)can be recruited to regulate the transcriptional activity of p21WAF1/CIP1 promoter in the breast cancer MCF-7 cells, and to clarify the molecular mechanism of suberoylanilide hydroxamic acid(SAHA) and leptin regulating p21WAF1/CIP1 promoter function.Methods The breast cancer MCF-7 cells in logarithmic growth phase were starved with FBS free medium for 24 hours, and treated with 20 μmol·L-1 SAHA(SAHA group) or 0.625 nmol·L-1 leptin(Leptin group) for 24 hours.The cells that were cultured in complete RPMI 1640 medium without any treatment were assigned as control group(Basal group).The cell lysis was prepared and incubated respectively with anti-HDAC1 and anti-ERα antibody by chromatin-immunoprecipitation(ChIP) method overnight at 4℃.The DNA-ChIP was followed the manufacturer′s protocol for the assay.DNA fragments binding anti-HDAC1 and anti-ERα antibody were gathered and purified.The relative expression level of DNA fragments from TSS to the upstream of the p21WAF1/CIP1 promoter region(+2~-4 000 bp) binding with antibody was detected by real-time PCR and analyzed by 2-△△CT method.Results In basal group, HDAC1 and ERα had high affinity with the f1 and f8 fragments of p21WAF1/CIP1 promoter compared to the f4 fragment.In SAHA group, the binding ability of HDAC1 and ERα to the f1 and f8 fragments of p21WAF1/CIP1 promoter was significantly lower than that of the control, while reversing to reach the peak after leptin treatment.Conclusions HDAC1 and ERα can be recruited to p21WAF1/CIP1 promoter by the cell proliferation signal during the proliferation of breast cancer MCF-7 cells.The DNA f1(from 0 to-400 bp) and f8(from-2 800 to-3 200 bp) fragment in the upstream of p21WAF1/CIP1 promoter are the target functional region for the binding with HDAC1and ERα.

To clarify the molecular mechanism of SAHA in the cell proliferation of ER-positive breast cancer cell line MCF-7 induced by leptin. Methods Human breast cancer cell MCF-7 was incubated with SAHA and/or leptin, and cell viability, apoptosis and cell cy-cle of MCF-7 cells were detected by Muse Cell Analy-zer. The expression of proteins related with apoptosis was determined by apotosis antibody array. Results Real-time cell proliferation assays indicated that the in-duction effect of leptin for MCF-7 cells reached the peak at a concentration of 0. 625 nmol · L-1 . SAHA reduced the viability of MCF-7 cells, induced G0/G1 phase arrest in the cell cycle, and triggered the apopto-sis. Meanwhile, SAHA significantly induced the pro-tein expressions of some apoptotic factors, including Bax, Caspase-3, TRAIL DR5, p21CIP1, and inhibited the expressions of Claspin, Clusterin, x-linked inhibi-tor of apoptosis protein(XIAP) and survivin. Howev-er, leptin had reverse effects on the related expression of the proteins. Conclusion The effects of cell prolif-eration by leptin and SAHA treatment in breast cancer ER positive cell line MCF-7 may involve in the activa-tion of apoptosis pathway, in particular with releasing of Caspase-3 trigged by endogenous mitochondrial ap-optosis pathway.

Objective To compare the ischemia-modified albumin(IMA)level between T2DM and AMI,and to discuss the possible mechanism and pathology process.Methods According to the decrease of power of albumin binding cobalt along with the increase of ischemia-modified albumin,the automated continuous detection of free cobalt in reactive system were conducted using the 480nm wavelength and the results were reported as absorbance unit(ABSU)and coloration rate of free cobalt.Results There were significant differences in serum free cobalt between the control and groups of acute coronary syndrome(ACS)and type 2 diabetes and between ACS and type 2 diabetes(P

Objective To screen the Doc 1R gene recombinant plasmid by use of colony PCR. Method The recombinant colonies were transfered into the PCR reaction mixture. The PCR primers were used for constructing mouse Doc 1R genomic sequence. Result Among the 5, 3 positive strips in the size of 1 500 bp were visible, which were the same as the Doc 1R gene fractions in terms of their sizes were screened as positive clones. The positive colony were further confirmed by double digestion and DNA sequencing. Conclusion Colony PCR is a simple, efficient and reliable technique for screening the recombinant.