Objective To investigate the apoptosis of cell line HL-60 induced by matrine and its possible mechanism. Methods CCK-8 assay was used to observe the proliferation inhibitation of HL-60 cells treated with matrine. FCM was applied to detect apoptotic cells and mitoehondrial transmembrane potential.Spectrophotometry was used to detect the activity of Caspase-9. Results Matrine (0.25-2.0 mg/ml) could significantly inhibit proliferation of HL-60 cells (F=67.83, P ＜0.05). Ater 48 hours, the apoptosis rate increased obviously in dose dependence (t = 4.685, 6.300. 9.641, 6.786, P ＜0.05). Within 48 hours, the mitochondrial transmembrance potential of HL-60 cells treated with matrine (1.0 mg/ml) gradually decreased (F = 54.83, P ＜0.05), and the activation of Caspase-9 gradually decreased in time dependence (F = 72.31,P ＜0.05). Conclusion Matrine can induce apoptosis of HL-60 cells by reducing mitochondrial transmembrance potential and activating Caspase-9.

AIM: To establish the method of determining cinobufagin and resibufogenin in Xinkening Capsules(Venenum Bufonis,etc.). METHODS: HPLC was applied to determining cinobufagin and resibufogenin.The determination was carried out using an ODS column with a solvent system: methanol-water(50∶40).The flow rate was 1.0 mL/min,detection wavelength was at 296 nm. RESULTS: The linear ranges of cinobufagin and resibufogenin were 3.5 ?g-0.28 ?g and 3.75 ?g-0.30 ?g,respectively.The average recoveries of cinobufagin and resibufogenin were 99.86%(RSD=1.8%,n=5),and 99.84%(RSD=1.2%,n=5),respectively. CONCLUSION: This method is simple,accurate with strong specificity and can be used to control the quality of Xinkening Capsules.

Objective To observe the therapeutic effects and side effects of FLAG regimen(fludarabine, Ara-C, and granulocyte-colony stimulating factor( C-CSF) for refractory or relapsed acute myeloid leukemia( AML). Methods Twenty-six AML were treated with fludarabine plus intermediate-dose Ara-C and G-CSF,of whom 15 cases belonged to refractory and 11 cases belonged to relapsed. Results After two courses of treatment, 14 cases were completely relieved (53. 8% ) and 5 cases were partially relieved (19.2% ). The overall effective rates was 73.1%. The main side effects were severe myelosuppression and non-hematological toxicity was mild. Conclusion FLAG regimen was very effective for refractory or relapsed acute myeloid leukemia and was well tolerated. The treatment-related mortality rate was low,so it provided a treatment choice for these patients.

Objective To evaluate the effect of Da Vinci surgical system for the treatment of hepatopancreatobiliary diseases.Methods The clinical data of 29 patients with hepatopancreatobiliary diseases who had undergone operations with Da Vinci surgical system from March to November 2009 at the General Hospital of PLA were retrospectively analyzed.Results The operations were successfully done on 28 patients,except 1 patient was converted to open pancreaticoenterostomy.The total operation time was(339±149)minutes,and the time for operations done with Da Vinci surgical system was(285±117)minutes.The postoperative bowl movement recovery time was(33±21)hours,and the length of postoperative hospital stay was(8±6)days.No blood transfusion was needed.Three patients had postoperative complications and were cured by conservative treatment.Conclusion Laparoscopic operations for hepatopancreatobiliary diseases can be applied with the help of the threedimensional imaging system and flexible surgical instruments of the Da Vinci surgical system,and its superiority is more obvious when applied for intractable hepatopancreatobiliary diseases.

Objective To investigate and cure major complications after percutaneous transhepastic biliary drainage(PTBD).Methods The clinical data of 13 major complications after PTBD were retrospectively analyzed,5 complications were acute and the other 8 complications were delayed.Two cases were dealed with intervention.and operations were performed for the other 11 patients immediately.Results Among the 7 patients who received one-stage operation,3 patients were accompanied with acute kidney failure,and 2 patients were died.Two patients who received the second-stage operation recovered snccessfully.Two patients who surrendered were surrived 3 and 8 months respectively.Conclusions It is difficult to deal with the major complications after PTBD,and the main cause of postoperative death is acute kidney failure.It will be helpful to deal with the primary disease on the second-stage.

Objective: Cell cycle has recently become more appealing as a new target of anti-carcinogen-ic agent. Diallyl disulfide (DADS) inhibits growth and induces call cycle G_2/M arrest in human gastric cancer BGC823 cells. Cell division cycle protein 25C (Cdc25C) and CyclinB1 expression are involved in G_2/M arrest.However, mechanisms of G_2/M arrest are not yet fully understood. The aim of this study was to elucidate the mechanism of cell cycle G_2/M arrest in human gastric cancer BGC823 cells induced by DADS. Methods: The expression of chk1 and Chk2 mRNA associated with cell cycle arrest of BGC823 cells after the induction with DADS for 1 or 2 days was detected by RT-PCR. The protein expression of cycle-related proteins ATM-RAD3-related gene (ATR), checkpoint kinase1 (Chk1), checkpoint kinase 2 (Chk2), P-ATR, P-Chk1 and P-Chk2 was measured by Western blot. Interaction between Chk1/2 and Cdc25C was analyzed by immuno-precipitation. Results: After the cells were treated with 15 mg/L DADS for 1 or 2 days, the expression of Chk1 and Chk2 mRNA was not significantly different from that in untreated cells (P>0.05). Western blot analysis showed that the expression of total Chk1 and Chk2 treated with 15 mg/L DADS was not significantly different from that in untreated cells. But phospho-chk1 showed a significant increase after stimulation with 15 mg/L DADS for 2h to 12h and continued to increase gradually as time went on (P<0.05). Phospho-Chk2 showed a eak expression and a weaker expression after stimulation with DADS, but the changes were not statistically significant (P>0.05). Addition of 15 mg/L DADS to BGC823 cells for 15 rain to 120 min resulted in an increase in phospho-ATR expression, whereas no changes were found in ATR expression (P<0.05). The Chk1 Ab in-creasingly precipitated Cdc25C in BGC823 cells treated with DADS (P<0.05). In contrast, Chk2 Ab failed to change precipitation with Cdc25C by DADS (P>0.05). Conclusion: Activation of chk1 was involved in cell cy-cle G_2/M arrest in BGC823 cells treated with DADS. Cell cycle G_2/M arrest by DADS is associated with phos-phorylation of several cell cycle regulatory proteins including ATR and Chk1 which regulate expression of Cdc25C.

Objective To investigate the distribution patterns of mosquitoes and mosquito-borne viruses in Dehong prefecture,Yunnan province,China. Methods Mosquito samples were collected using the mosquito traps from five counties of Dehong prefecture on July,2007 and 2010. Mosquitoes were cell cultured for viral isolation,and positive isolates were identified using RT-PCR and sequence analysis. Results A total of 43 634 mosquitoes comprised of 29 species representing six genera were collected. Culex tritaeriorhynchus and Anopheles sinensis comprised 78.69% and 14.77% of the total. Six strains of viruses were isolated from the mosquito pools. RT-PCR and phylogenetic analysis revealed three strains from Cx. tritaeriorhynchus,identified as genotypeⅠJapanese encephalitis virus(JEV). One strain was identified from Cx. tritaeriorhynchus,as Getah virus (GETV). Two strains isolated from Cx. tritaeriorhynchus and Anopheles vagus were identified as Culex pipiens pallens Densovirus(CppDNV). Conclusion Cx. tritaeriorhynchus had been the major species of mosquitoes and mainly transmitting vector of mosquito-borne viruses in Dehong prefecture. GenotypeⅠJEV,GETV and CppDNV were the vectors causing transmission of mosquitoe-borne diseases in this area. Data from phylogenetic analysis showed that these newly discovered isolates seemed to have had close relationship with those viruses previously circulating in Yunnan and other provinces of China.

Objective To ensure the effectiveness and improve the capacity of laboratories in the measurement of γ spectra of radionuclides. Methods A statistical analysis was performed using relative deviation, accuracy, precision, and relative combined uncertainty for the nationwide assessment of radionuclide γ spectrum measurement. Results In the assessment from 2018 to 2020, our laboratory showed qualified or above results, and the performance in 2019 was excellent. The maximum relative deviation of 11 measurements from 2018 to 2020 was 18.01%. The assessment showed |Z_test|≤1 and U_test≤2.58 in 2018, |Z_test|≤1 and U_test≤1 in 2019, and U_test≤1 and U_rel≤20% in 2020. Conclusion Our laboratory employs the correct method for radionuclide γ spectrum measurement, and the analysis data are accurate and reliable.

Objective To improve the analysis of ¹³⁴Cs, ¹³⁷Cs, and ⁶⁰Co in aerosol samples by the national key radiation environment laboratories. Methods Intercomparison of analysis results of ¹³⁴Cs, ¹³⁷Cs, and ⁶⁰Co in standard aerosol samples was performed among the national key radiation environment laboratories according to Gamma spectrometry method of analyzing radionuclides in biological samples (GB/T 16145-1995 ), and the intercomparison results were evaluated by the standard deviation. Results Six laboratories were involved in the intercomparison. For ¹³⁴Cs, 50% of the laboratories showed a relative deviation less than 10%, and 50% showed a relative deviation of 10%-20%. For ¹³⁷Cs, 33.3% of the laboratories showed a relative deviation less than 10%, and 76.7% showed a relative deviation of 10%-20%. For ⁶⁰Cs, all laboratories showed a relative deviation less than 10%. The overall intercomparison results were acceptable. Conclusion The laboratories in this intercomparison show generally good results.

OBJECTIVE：To excavate the safety warning signals induced by azole antifungal agents ，including fluconazole ， ketoconazole，itraconazole and voriconazole after marketing ，and to provide references for rational drug use in the clinic. METHODS：Reporting odds ratio （ROR）data mining algorithm was used to investigate signals of adverse drug event （ADE）for fluconazole，ketoconazole，itraconazole and voriconazole from FDA Adverse Event Reporting System （FAERS）during January 1st，2004 to March 30th，2019. ROR data mining method was used to excavate the ADR signals of the drugs ，and main ADR involved in the safety information of azole antifungal agents instructions were analyzed. RESULTS ：A total of 27 831，5 712， 5 381 and 11 333 reports were picked out for fluconazole ，ketoconazole，itraconazole and voriconazole ，respectively. All of these drugs had exhibited high-risk signals detection by ROR ，including medical examination ，blood and lymphatic system disorders ， renal and urinary disorders ，endocrine diseases ，hepatobiliary disorders. The hepatotoxic-related ADR signals were mainly concentrated in fluconazole and voriconazole （fluconazole ROR ＝6.51，voriconazole ROR ＝14.65）；ADR detection results of Cushing’s-like syndrome （ROR＝24.86） and adrenal suppression （ROR＝44.06） by itraconazole showed high-risk signals ； ketoconazole and itraconazole had showed a strong ADR signal in adrenocortical dysfunction （ketoconazole ROR ＝15.64， itraconazole ROR ＝23.26），and the signal intensity of ketoconazole （ROR＝2.81）in skin and subcutaneous tissue disorders was significantly higher than that of other drugs . In addition ，hemorrhagic cystitis caused by fluconazole，itraconazole and voriconazole were not included in the drug instructions （fluconazole ROR ＝17.73，itraconazole ROR ＝31.43，voriconazole ROR ＝17.06）； netted green spot caused by fluconazole （ROR＝10.50）were not included in the drug instructions . CONCLUSIONS：Clinical staff should pay more attention to the differences in serious ADR related to fluconazole ，ketoconazole，itraconazole and voriconazole ； particularly some ADRs not mentioned in the drug instructions but have high incidence such as hemorrhagic cystitis caused by fluconazole，itraconazole，voriconazole and netted green spot caused by fluconazole ，as well as ADRs mentioned in the drug instructions but have abnormally high signal ，such as Cushing ’s-like syndrome and adrenal suppression caused by itraconazole .