Objective To investigate the relation between demethylation effect and apoptosis of valproate (VPA) in primary acute lymphoblastic leukemia (ALL) cells in vitro.Methods 10 cases of ALL patients was choosed to acquire leukemia cells.Cell growth curve were assessed by the MTT assay,the apoptosist of primary ALL was analyzed with DNA Ladder and Annexin-V-FITC/PI by flow cytometry.The expression methylation level of p15 was detected by hn-MSPCR,and p15mRNA were detected by RT-PCR.All dates was analyzed by SPSS16.0.Results The 50 ％ inhibition rate of VPA were 1.898 mmol/L to primary ALL cells assayed by MTT respectively.DNA ladder showed the apoptosis of primary ALL cells increased by adding VPA dose.Annexin-V-FITC/PI tests showed that the apoptosis percentage of primary ALL cells were (0.44±0.04) ％ in control group,(5.80±0.65) ％ in 1.0 mmol/L VPA group,(48.46±2.49) ％ in 2.0 mmol/L VPA group,(76.45±2.98) ％ in 4.0 mmol/L VPA group,the apoptosis percentage increased significantly (P ＜ 0.05).The demethylation of p15 INK4B gene decreased by adding VPA dose,the expression of p15 mRNA expression increased significantly compared with control group by RT-PCR (P ＜ 0.05).Conclusion It is found that VPA could induce demethylation of p15 INK4B gene,which could upregulate the p15 mRNA expression,due to the apoptosis of primary ALL cells.

Objective To investigate the inhibition of proliferation and the regulation of histone acetylation modification in Jurkat cells treated by sodium valproate(VPA). Methods Jurkat cells were treated with VPA.Cell proliferation was determined by CCK-8 assay, and cell cycle were analyzed by flow cytometry (FCM). mRNA of HDAC1 was detected by semi-quantitative RT-PCR, and protein expression of HDAC1 and acetylation of histone H3, H4 was examined by Western blotting. Results VPA inhibited the proliferation of Jurkat cells in concentration-and time-dependent manners. After exposure to VPA in different concentrations for 48h,cell cycle was arrested obviously at G0/G1 phase (P <0.05), and with increasing concentration, the percentage of G0/G1 phase cells was increased and that of S phase were decreased. HDAC1 mRNA expression were inhibited with the increasing concentration of VPA. The protein level of HDAC1 was down-regulated, while acetylation of histone H3、H4 was up-regulated in Jurkat cells by VPA. Conclusion VPA can inhibit proliferation of Jurkat cells and induce G0/G1 phase arrest. The mechanism may be that VPA increase acetylation of histone H3/H4 by inhibiting expressions of HDAC1 gene.

Objective To explore the characteristics of hemi-nested methylation specific polymerase chain reaction (hn-MSP) and to find out the possible relationship between patterns of methylation or deletion and the developmet of adult acute lymphoblastic leukemia(ALL).Methods hn-MSP and bisulfit-sequencing PCR (BSP) were designed and adopted to analyze p15 gene methylation or deletion patterns in 25 adult ALL patients,malignant hematopathy cell lines and normal lymphocytes. hn-MSP and BSP products were cloned and sequenced.The sensitivity and specificity of hn-MSP were also analized.Results The sequencing results of hn-MSP and BSP products were consistent, and the sensitivity of detection of p15 methylation was up to 1.0×10-5.17 adult ALL patients (68 ％) were p15 gene hypermethylation and 3 patients were with deletion of p15 gene exon 1.There were no hypermethylation or deletion in the 10 controls.Conclusions The detection rate of p15 methylation in many tumors,especially in adult ALL,is frequent high.hn-MSP is highly sensitive and specific in analyzing p15 methylation.

Objective To explore the mechanism of apoptosis induced by diacetyl hexamethylene diamine(CAHB)in adult acute lymphoblastic leukemia Jurkat cells,and to provide theoretical basis for the clinical application of CAHB.Methods Annexin V+/PI-cell rate of Jurkat cells after CAHB induction was analyzed by flow cytometry.The Annexin V+/PI-cell rate was observed after treatment with caspase-9 inhibitor Z-LEHD-FMK.The expressions of apoptosis-related proteins caspase-8,caspase-9 and caspase-3 were observed by Western blotting.Results After CAHB induction,Jurkat cells were reduced in size,cell membrane crinkling,chromatin thickening,nuclear pyknosis or fragmentation,etc.Typical apoptotic bodies could be seen.CAHB induced Jurkat cell apoptosis by activating caspase-9 and caspase-3 in a dose-effect and time-dependent manner.Caspase-9 inhibitors could inhibit apoptosis induction of CAHB to a certain extent.Conclusion CAHB induced Jurkat cell apoptosis was related to caspase-9 and caspase-3 activation.

Chronic refractory wound (CRW) is one of the most challengeable issues in clinic due to complex pathogenesis, long course of disease and poor prognosis. Experts need to conduct systematic summary for the diagnosis and treatment of CRW due to complex pathogenesis and poor prognosis, and standard guidelines for the diagnosis and treatment of CRW should be created. The Guideline forthe diagnosis and treatment of chronic refractory wounds in orthopedic trauma patients ( version 2023) was created by the expert group organized by the Chinese Association of Orthopedic Surgeons, Chinese Orthopedic Association, Chinese Society of Traumatology, and Trauma Orthopedics and Multiple Traumatology Group of Emergency Resuscitation Committee of Chinese Medical Doctor Association after the clinical problems were chosen based on demand-driven principles and principles of evidence-based medicine. The guideline systematically elaborated CRW from aspects of the epidemiology, diagnosis, treatment, postoperative management, complication prevention and comorbidity management, and rehabilitation and health education, and 9 recommendations were finally proposed to provide a reliable clinical reference for the diagnosis and treatment of CRW.