Objective To observe the cell proliferation inhibition of DNA methyltransferase (DNMT) inhibitors decitabine (DAC) combined with daunorubicin (DNR) in human leukemia cell line HL-60.Methods The effects of DNR and DAC were examined in HL-60 cells by cell viability using MTT method,and cell death using flow cytometric (FCM).Results DAC,DNR single drug application showed their effects on cell proliferation was dependent of dose and time,the inhibition effect of combined treatment group was much clearer [inhibition ratio of 72 hours was (80.23±1.71) ％,P ＜ 0.001].The highest apoptosis rate was at 5.0 μmol/L DAC combined with 1.0 μmol/L DNR for 72 hours,which was statistic significant (F =30.199,P ＜ 0.001).Combinations of different concentrations of DAC and DNR increased expression of PTEN mRNA in concentration-dependent manner,which was significantly higher than the control group and DNR single drug group (F =578.218,P ＜ 0.001).Conclusions DAC can significantly inhibit the proliferation of HL-60 cells and induce apoptosis,synergistic effect can be observed when DAC combined with DNR.The underlying mechanism can be due to DAC demethylation effect to increase PTEN mRNA expression.

[Objective] To detect the expression levels of connexin43(Cx43),P-gp and COX-2 in bone marrow of patients with acute leukemia (AL),and investigate their relationship with the pathogenesis,prognosis and resistance of this condition.[Methods]Using SYBR green real-time quantitativereverse transcription polymerase chain reaction (SYBR-RT-PCR),the expression of Cx43,P-gp and COX-2 mRNA were detected in 77 AL patients at different phases,including 36 initially-treated,20 complete-remission (CR)and 20 relapsed.A follow-up was done in those initially lreated.[Results]Expression levels of Cx43,P-gp and COX-2 mRNA from initially-treated were 0.52±0.57,1.42 ±1.06,1.14±0.95;those from relapse AL patients were 0.20±0.40,2.29 ±1.11,1.69±0.81,respectively,those from CR patients were 0.95±0.37,0.93±-0.73,0.79±0.58,respectively,those from normal patients were 1.16 ±0.67,0.86±0.63,0.61±0.57.Expression levels of Cx43 mRNA in initially-treated and relapse AL patients were significantly lower than those in normal patients and CR patients(initially-treated P were 0.001,0.005;relapse patients were P＜0.001),while the comparison between normal patients and CR patients showed no statistical significance(P=0.185).Cx43mRNA expression level was negatively correlated with P-gp and COX-2 mRNA,and a negative correlation was noted of both two expressions in initially-treated and relapse groups (Cx43 mRNA and P-gp mRNA r were -0.471,-0.362,-0.526;Cx43 mRNA and COX-2 mRNA r were -0.479,-0.344,-0.471).A follow-up of four months was conducted in 36 initially-treated patients,eight of them died.The expression Cx43 in those who died was lower than that who survived.The difference was significant(t=2.16,P=0.042).[Conclusion] Cx43mRNA expression levels of initially-treated and relapse AL patients were lower than those in normal patients and CR patients,P-gp and COX-2 mRNA expression levels of initially-treated and relapse AL patients were higher than those in normal patients and CR patients.Cx43 expre-ssion probably is a favorable prognostic factor.Cx43 is participation in the genesis,development and drug resistance of AL.

Objective To observe the protein expression of grow thassociated protein-43 (GAP-43) in mid-brain ventral tegmental area in morphine withdrawal rats at different time, and to evaluate the effect of GAP-43 on morphine withdrawal memory. Methods Rat models of morphine dependent 1 week, 2 weeks and 4 weeks were established by morphine hydrochloride intraperitoneal injection with increasing doses to establish natural withdrawal. The protein expression of GAP-43 in midbrain ventral tegmental area was observed by im munohistochemical staining and the results were analyzed by Im age-Pro Plus 5.1 im-age analysis system . Results With prolongation of dependent time, the expression of GAP-43 was de-creased then increased in midbrain ventral tegmental area . Conclusion GAP-43 could play arole in morphine withdrawal memory in midbrain ventral tegmental area.

Objective To observe hepatocellular apoptosis and inflammatory cytokines expression and their mechanisms after paraquat poisoning in rat.Methods Forty Wistar rats were divided into control group (n =8) and model group (n =32) by random number table.Rats in model group were intraperitoneally injected with 30 mg/kg 20％ paraquat concentrate,while those in control group were injected with normal saline.0.5,1,3,7 days after reproduction of the model,8 rats were sacrificed,and blood was collected from inferior vena cava and hepatic tissue was harvested.The serum levels of interleukin-1β (IL-1 β) and tumor necrosis factor-α (TNF-α) were determined by enzyme-linked immunosorbent assay (ELISA).The mRNA expressions of IL-1β,TNF-α,inducible nitric oxide synthase (iNOS) and p53 were determined by reverse transcription-polymerase chain reaction (RT-PCR).Cysteine-containing aspartate-specific proteases (caspase-3,-8,-9,-12) activity in hepatic tissue was determined on the 3rd day with chromogenic substrate method.The liver histopathological changes were observed after hematoxylin-eosin (HE) staining.Results In model group,hepatic tissue showed extensive necrosis with inflammatory cell infiltration in time dependant manner.Serum IL-1β and TNF-α levels were significantly higher in model group half a day after reproduction than those in control group [IL-1β (ng/L):220.13 ± 69.74 vs.0.14 ± 0.03,TNF-α (ng/L):102.66 ± 26.43 vs.0.16 ± 0.02,P＜ 0.01 and P＜0.05],and peaked on the 3rd day and 1st day [IL-1β:(423.72 ± 153.11) ng/L,TNF-α:(690.35 ± 229.64) ng/L].They then decreased gradually,but were still significantly higher than those in control group on the 7th day [IL-1 β:(357.47 ± 87.28) ng/L,TNF-α:(12.39 ± 5.06) ng/L,both P＜0.05].The contents of IL-1β,TNF-α and iNOS mRNA expressions in hepatic tissue were significantly higher than those in control group,and the highest values were seen on the 1st day,the 1st day,and the 3rd day [IL-1β mRNA (gray value):1.569 ± 0.057 vs.0.123 ± 0.016,TNF-α mRNA (gray value):0.683 ± 0.077 vs.0.261 ± 0.025,iNOS mRNA (gray value):3.259 ± 0.135 vs.0.002 ±0.001,P＜0.05 or P＜0.01].There was no difference in p53 mRNA expression between model group and control group at early stage,and both of them showed low expression,and p53 mRNA expression was significantly higher in model group on the 7th day (gray value:2.959 ± 0.086 vs.0.263 ± 0.032,P＜0.01).In model group,caspase activity (pmol/mg) in liver tissue were significantly higher on the 3rd day than those in control group (caspase-3:857.25 ± 309.26 vs.169.73 ± 48.21,caspase-8:199.18 ± 61.41 vs.32.26 ± 11.09,caspase-9:321.62 ± 80.73 vs.90.38 ± 29.76,caspase-12:413.13 ± 89.77 vs.26.73 ± 9.86,all P＜0.01).Conclusion Paraquat can cause acute liver injury in rats,with caspase-3,-8,-9,-12 activities markedly enhanced,and liver injury may be associated with an early high expression of TNF-α,iNOS and p53 gene.

Objective To observe hepatocellular apoptosis and inflammatory cytokines expression and their mechanisms for lipopolysaeeharide (LPS)-induced acute liver failure in D-ga|actosamine (D-GalN)-sensitized rats. Methods Fifty-six rats were randomly divided into following groups: 0, 1, 2, 4, 6, 24 and 48 hours. 0 hour group served as control group and the rest did as treatment groups. The rats in the treatment groups received intraperitoneal injections of LPS (50 ng/g) and D-GaIN (300 μg/g) dissolved in 1 mL sterile 0.9% sodium chloride solution, while the rats in control group were treated with 1 mL sterile 0.9% sodium chloride solution only. The rats were sacrificed in the corresponding time points and their sera and liver tissues were collected. Liver tissues were fixed and stained with hematoxylin and eosin for optical microscopy examination. The serum cytokine expressions were detected by enzyme-linked immunosorbent assay (ELISA). The expressions of tumor necrosing factor (TNF)-α, interleukin (IL)-1β, inducible nitric oxide synthase (iNOS) and p53 gene were detected by reverse transcriptase-polymerase chain reaction, and the 24 hours treated rats liver Caspase-3,8,9,12 activity were detected by chromogenie substrate method. Data for the experiments were expressed as x±s, and differences among means were compared using the analysis of variance. Results After drug treatment, liver tissues showed piecemeal necrosis and inflammatory cell infiltration, which significantly increased from 6 hours, 24 hours to 48 hours. The 1 hour treatment group with the highest concentration of TNF-α (727. 8 ± 261. 3) ng/L were significantly higher than the control group and other treatment groups(F= 49.82, P<0.01), 2 hours treatment group (156.4 ± 52.2) ng/L was significantly lower than the 1 hour group, but significantly higher than the control group (F = 30. 23, P< 0.01 ). But serum concentrations of IL-1β gradually increased, reaching the highest level in 24 hours group (360.5±121.6)ng/L (F= 18. 61, P<0. 01). Liver Caspase-3,8,9, 12 activity in 24 hours treatment group was significantly higher than in the control group (F= 84.96, P<0.01). The mRNA expression of iNOS gene, which were not detected in normal controls, reached the peak at 6 hours group after drug treatment and notably dropped in 24 hours and 48 hours groups(F=34.07,P<0.01), p53 gene expression significantly upregulated at 24 hours and 48 hours groups(F=37.43,P<0.01). TNF-α and IL-1β gene expression in the treatment group were higher than in the control group(F=2.94,P<0.05), and both reached the peak at 1 hour treatment group. Conclusions Acute liver failure can be induced by low dose LPS in D-GaiN-sensitized rats. One of the features changes is that Caspase-3,8,9,12 activities are markedly enhanced, and the occurrence of liver injury may be associated with the early high expression of TNF-α, iNOS and p53 gene.

Objective To observe the expression of discs large hom olog 4 (DLG4) protein in hippocam-pus, am ygdala and frontal cortex of rats and evaluate postsynaptic density in heroin dependence. Meth-ods The rat heroin dependent m odel was established by increasing intraperitoneal injection of heroin. DLG4 proteins in hippocam pus, am ygdala and frontal cortex of heroin dependent 9, 18, 36 days rats w ere detected with im munohistochem ical staining and com pared with that in the control group. Results DLG4 proteins in hippocam pus, am ygdala and frontal cortex w ere gradually reduced with extension of heroin dependent tim e. Conclusion Heroin dependence can affect postsynaptic density of hippocam pus, am ygdala and frontal cortex. The changes becom e m ore apparent with extension of heroin dependence tim e.

Objective To explore the effect of training mode based on action learning on improving the practicing ability of hospital pharmacists.Methods Thirty pharmacists who received training from September 2022 to December 2023 at Panzhihua Central Hospital were randomly divided into an education reform group(16 cases)and a routine group(14 cases).The education reform group adopted a routine teaching method based on action learning,while the routine group adopted a routine teaching method.The differences between the two groups of pharmacists in theoretical knowledge,practical operation,pharmaceutical services,emergency response,and comprehensive quality were compared.Results The pharmacists in the education reform group were better than the routine group in prescription review,clinical medication analysis,pharmaceutical services,emergency response,andcomprehensive quality.The difference was statistically significant(P＜0.05).Conclusion The teaching model based on action learning can effectively enhance the higher order thinking ability of pharmacists and help them better apply medical knowledge and skills to serve patients and physicians.

Objective: To investigate the efficacy and safety of IA regimen which contains idarubicin (IDA) 8 mg/m², 10 mg/m² or 12 mg/m² as induction chemotherapy for adult patients with de-novo acute myeloid leukemia (AML) . Methods: A total of 1 215 newly diagnosed adult AML patients, ranging from May 2011 to March 2015 in the First Affiliated Hospital of Soochow University and other 36 clinical blood centers in China were enrolled in the multicenter, single-blind, non-randomized, clinical controlled study. To compare the response rate of complete remission (CR) , adverse events between different dose idarubicin combined with cytarabine (100 mg/m²) as induction chemotherapy in newly diagnosed patients of adult AML. Results: Of 1 207 evaluable AML patients were assigned to this analysis of CR rate. The CR rates of IDA 8 mg/m² group, IDA 10 mg/m² group and IDA 12 mg/m² group were 73.6% (215/292) , 84.1% (662/787) and 86.7% (111/128) , respectively (P<0.001) . After adjusted for age, blast ratio of bone marrow, FAB classification and risk stratification, the odds ratios (95% CI) of IDA 10 mg/m² group and IDA 12 mg/m² group were 0.49 (0.34-0.70) and 0.36 (0.18-0.71) , as compared with the IDA 8 mg/m² group (P<0.001, P=0.003) . In the intermediate and favorable groups, CR rates was 76.5% (163/213) , 86.9% (506/582) and 86.1% (68/79) in different doses of IDA (P=0.007) . Interestingly, IA regimen with IDA 10 mg/m² was the only beneficial factor affecting CR in this group after adjusted for age, blast ratio of bone marrow and FAB classification[OR=0.47 (95% CI 0.31-0.71) , P<0.001]. CR rates in adverse group was 50.0% (18/36) , 60.6% (43/71) and 81.8% (18/22) respectively (P=0.089) . However, the odds ratios (95% CI) of IDA 12 mg/m² when compared with the IDA 8 mg/m² was 0.22 (0.06-0.80) , after adjusted for age, blast ratio of bone marrow and FAB classification. The median time (days) of neutrophil count less than 0.5×10⁹/L in IDA 8 mg/m² group, IDA 10 mg/m² group and IDA 12 mg/m² group were 14 (11-18) , 15 (11-20) and 18 (14-22) , respectively (P=0.012) and of platelet count lower than 20×10⁹/L were 14 (7-17) , 15 (11-20) and 17 (15-21) , respectively (P=0.001) . The incidences of lung infection in the three groups were 9.8%, 13.5% and 25.2%, respectively (P<0.001) . Conclusions For young adult patients (aged 18-60 years) with AML in China, intensifying induction therapy with idarubicin 10 mg/m² is clinically superior to IDA 8 mg/m² and IDA 12 mg/m² in favorable intermediate AML subgroup. However, idarubicin 12 mg/m² is more suitable to adverse AML subgroup.