Objective To analyze the safety of high dose methotrexate (HD-MTX) in the treatment of children with acute lymphoblastic leukemia (ALL) after 36 hours and 42 hours using leucovorin (CF) rescue. Methods A total of 137 childhood with acute lymploblastic leukemia(ALL) in our hospital from September 2012 to December 2015 were analysed in this study, 137 children with ALL were randomly divided into group A (n=69) and group B (n=68) according to the serial number (odd number and odd number). The total number of treatment times was 454 times. Among them, the rescue time of group A was 36 hours, there were 224 cases, and the rescue time of group B was 42 hours, 230 times.The plasma drug concentration, delayed excretion rate, the total dose of leucovorin, the total dose of methotrexate and the incidence of side effects were observed in group A and group B at 24, 48 and 96 hours. Results There was no significant difference in plasma concentration, delay of excretion and incidence of side effects between 2 groups of methotrexate, 24, 48 and 96 hours. The total dose of methotrexate/methotrexate in 2 groups was statistically significant (P＜0.05). Conclusion When the rescue time of leucovorin was 42h, the treatment of acute lymphoblastic leukemia with high-dose methotrexate was the best.

OBJECTIVE:To establish the RP-HPLC method for determination of ginsenoside Rg1 in Aidi injection. METHODS:The determination was performed on Lichrospher C18(250 mm?4.6 mm,5 ?m) with column temperature kept at 35 ℃. The mobile phase consisted of water -acetonitrile (gradient elution) at a flow rate of 1.0 mL?min-1.The detection wavelength was set at 203 nm. RESULTS:The linear range of ginsenoside Rg1 was 5.0～250.0 ?g?mL-1(r=0.999 5),and the average recovery was 97.43%(RSD=1.81%,n=6). CONCLUSION:The method is simple,rapid and accurate,and it is suitable for the quality control of Aidi injection.

Objective:To evaluate the efficacy and pharmacoeconomics of rhIL-11(Ⅰ) and rhTPO in the treatment of thrombocy-topenia caused by gemcitabine chemotherapy in lung cancer patients. Methods:A retrospective analysis was used. Totally 58 hospital-ized lung cancer patients who suffered thrombocytopenia caused by gemcitabine chemotherapy and treated with rhIL-11(Ⅰ) or rhTPO from June 2011 to June 2014 were involved in the study, and the efficacy and pharmacoeconomics of rhIL-11(Ⅰ) and rhTPO were e-valuated and compared. Results:The lowest platelet value after the chemotherapy in rhIL-11(Ⅰ) group was higher than that in rhTPO group (P<0. 01), and the platelet decrease time in rhIL-11(Ⅰ) group was shorter than that in rhTPO group (P<0. 01), while the platelet qualified rate of the two groups showed no statistically significant difference (P>0. 05). The results of cost-minimization anal-ysis showed that the average cost of rhIL-11(Ⅰ) group was lower than that of rhTPO group(P<0. 01), furthermore, the average cost of the patients with GP, GC or the other gemcitabine chemotherapy regimens in rhIL-11 (Ⅰ) group was lower than that in rhTPO group. Conclusion:The effect of rhIL-11 (Ⅰ) in the treatment of thrombocytopenia caused by gemcitabine based-chemotherapy in lung cancer patients is not inferior to that of rhTPO, and shows certain advantages in economic cost.

OBJECTIVETo develop a HPLC method for simultaneous determination of caffeic acid, rosmarinic acid, oleanolic acid and ursolic acid in Spica Prunellae.

METHODAfter ultrasonic extraction with 75% ethanol solution containing 1% formic acid, the ethanol-extract of Spica Prunellae was analyzed on an Elite SinoChrom ODS-AP column using gradient elution of 0.01% phosphoric acid (A) and acetonitrile (B) at a flow rate of 0.9-1.0 mL x min(-1). A wavelength switch program was used for detection at 330 nm (0-33 min) and 203 nm (33-40 min). The column temperature was set at 20 degrees C and injection volume was 50 microL.

RESULTThe calibration curves of all analytes were linear. The average recoveries were 93.7%-105.2% with RSDs not more than 4.5%. The contents of caffeic acid, rosmarinic acid, ursolic acid and oleanolic acid in Spica Prunellae were 0.0401-0.0968, 0.99-2.57, 0.243-0.556, 4.06-8.13 mg x g(-1), respectively.

CONCLUSIONThe described method is sensitive, convenient and accurate, and is suitable for the simultaneous determination of caffeic acid, rosmarinic acid, oleanolic acid and ursolic acid in Spica Prunellae.

Caffeic Acids ; analysis ; Calibration ; Chromatography, High Pressure Liquid ; methods ; Cinnamates ; analysis ; Depsides ; analysis ; Oleanolic Acid ; analysis ; Prunella ; chemistry ; Triterpenes ; analysis

The mollusicidal effect and toxicity to fish with META-Li in field were tested in mountain areas in Zhejiang Province. The snail control test, which had 3 groups including a META-Li group, niclosamide group and control group, were performed by the spraying method. The adjusted mortalities of snails in the META-Li group on the 3rd, 7th and 15th day were 89.76% , 87.98% and 94.10% , respectively, in the niclosamide group were 89.70% , 83.22% and 94.72% , respectively .and there was no significant difference between the two groups ( P > 0.05), but they were decreased obviously compared with the control group, which were 9.24% , 9.50% and 15.21% , respectively(P <0.05). The toxicity test to fish with META-Li included 3 groups, namely a high concentration META-Li group, low concentration META-Li group and control group, and none fish was dead. It is suggested META-Li has low toxicity to fish, and is suitable to snail control in aquiculture areas.

ObjectiveTo investigate the clinical characteristics,epidemiology of patients with severe fever with thrombocytopenia syndrome bunyavirus (SFTSV) infection and genetic sequences of SFTSV.MethodsClinical data of five cases of severe fever with thrombocytopenia syndrome (SFTS)from Zhoushan Hospital during May 2011 to July 2011 were retrospectively analyzed.SFTSV gene was amplified by polymerase chain reaction (PCR).CD3+ CD4+ and CD3+ CD8+T lymphocytes were detected by flow cytometry (FCM).The sequences of isolated SFTSV strains were compared with those in GenBank. ResultsThe symptoms of continuous high fever,sore muscles,enlarged superficial lymph nodes,abdominal pain,diarrhea with gastrointestinal hemorrhage were observed.The white blood cells,platelets and CD3+ CD4+ T lymphocytes were progressive decreased in acute phase with the minimum of (0.97-2.00) × 109/L,(12-42) × 109/L and 7.52％-20.39％,respectively.The SFTSV was isolated from the sera of two patients.The sequences were compared with SFTSV sequences in GenBank.The homology of RNA-dependent RNA polymerase gene was 96％ compared with BX-2010,L-WWG,LN3,JS4,SD4,HN6 and AH12; the glycoprotein gene was 94％ ; N protein gene was 95％ compared with JS4,SD4 and LN4.The homology of the above three genes between two isolates was 99％.ConclusionsOur results suggest that SFTSV is sporadic in Zhejiang Province which is probably from native epidemic focus.SFTS is progressive and severe with acute onset.Multiple organ dysfunction is common in severe eases.

OBJECTIVE:To study the inhibitory effect and mechanism of Inula helenium ethyl acetate extract(IHE)on prolif-eration of human pancreatic cancer Capan-2 cells. METHODS:MTT was used to determine the cell proliferation inhibition rate af-ter treated by 0,0.5,1,2,4,8 μg/mL IHE;clone formation test was used to observe the effects of 0,1,2 μg/mL IHE treating for 1 week on cell clone formation;Hoechest 33342 staining was used to observe the changes of nuclear morphology after treated by 0,2,4 μg/mL IHE for 48 h;flow cytometry was used to detect the cell apoptosis rate after treated by 0,4,8,16 μg/mL IHE for 48 h;JC-1 staining was used to observe the changes of intracellular mitochondrial membrane potential after treated by 0,4,8, 16 μg/mL IHE for 24 h;Western blot was used to detect the expressions of mitochondrial apoptosis-related proteins Bcl-2,Bax, Mcl-1,p53 up-regulated modulator of apoptosis (PUMA),and polymerase (PARP) after treated by 0,4,8,16 μg/mL IHE for 48 h. RESULTS:2,4,8 μg/mL IHE had obvious inhibitory effect on cell proliferation,showing concentration-dependent relation-ship,with IC50 of 6.6 μg/mL;1,2 μg/mL IHE can obviously inhibit the clone formation of cells;4 μg/mL IHE can obviously cause cell nuclear condensation;8,16 μg/mL IHE can obviously promote the cell apoptosis,and the cell apoptosis rate reached 45.53% after treated by 16 μg/mL IHE for 48 h;16 μg/mL IHE treating for 24 h can cause the decrease of 82.47% cells'mito-chondrial membrane potential;8 μg/mL IHE can obviously down-regulate the protein expressions of Bcl-2,Mcl-1,PUMA and PARP,and 16 μg/mL IHE can obviously down-regulate the expressions of Mcl-1 and PUMA. CONCLUSIONS:IHE may show its inhibitory effect on proliferation of human pancreatic cancer Capan-2 cells by causing the decrease of mitochondrial mem-brane potential in cells and down-regulating the protein expres-sions of Mcl-1 and PUMA to cause cell apoptosis.

OBJECTIVE：To realize refined management of tablets i n the inpatient pharmacy ，and to ensure the medication safety of patients. METHODS ：Based on intelligent pharmacy ，the dispensing and packaging process under the automated drug dispensing and packaging system （ADDPS）mode was optimized and modified ；PDA barcode scanning technology was applied in all links of taking ，dismantling and adding ，so as to realize the real-time tracking of batch number and inventory ，and improve the drug closed-loop management of tablets. The error rate ，staff consumption time and pharmacist/nurse satisfaction were compared before and after the process improvement. RESULTS ：After the process improvement ，the dispensing error rate was decreased from 1.637‰ before improvement to 0.082‰（P＜0.01）；the staff consumption time decreased from （7.52±0.33）h to （5.11±0.24）h （P＜0.01）；the pharmacist/nurse satisfaction increased from 66.5％ to 93.4％（P＜0.01）. CONCLUSIONS ：Based on ADDPS ，the application of PDA barcode scanning technology standardizes the tablets management of inpatient pharmacy ，supplements and improves the drug closed-loop information ，realizes batch number tracking and inventory management ，reduces the occurrence of tablet dispensing errors ，improves the work efficiency and satisfaction of pharmacists ，and ensures the safety of clinical medication.

OBJECTIVE： To investigate the inhibitory effect and potential mechanism of Brucein D （BD） combined with Taxol on the proliferation of human pancreatic cancer Capan-2 cells. METHODS： Using Capan-2 cells as object， the proliferations after treated with BD （5， 10， 15， 20 μmol/L）， Taxol （10， 20， 30， 40 nmol/L） and BD+Taxol （5 μmol/L+10 nmol/L， 10 μmol/L+20 nmol/L， 15 μmol/L+30 nmol/L， 20 μmol/L+40 nmol/L） for 48 h were determined by sulfonyl rhodamine B method. Survival rate of cells and combination index （CI） were calculated. The clone formation assay was performed to detect the formation of clonal colonies after treated with BD （20 μmol/L，hereinafter）， Taxol （40 nmol/L，hereinafter）、BD+Taxol （20 μmol/L+40 nmol/L，hereinafter） for 24 h. The rate of clone formation was calculated. DAPI method was used to observe the apoptosis of cells after treated with BD， Taxol and BD+Taxol for 24 h. Western blotting was used to detect the expression of apoptosis-related protein （Bcl-2， PARP， Caspase-3， Cleaved-caspase-3） after treated by BD， Taxol， BD+Taxol for 48 h and the expression of JNK and p-JNK after treated by BD， Taxol， BD+Taxol for 4， 6， 12 h. RESULTS： After treated with 10， 15 and 20 μmol/L BD， 20， 30 and 40 nmol/L Taxol or two-drug combination for 48 h， survival rates of cells were decreased significantly； the survival rate of drug combination group was significantly lower than the same dose of BD group and Taxol group （P＜0.05）. CI values of drug combination groups （BD 5 μmol/L+Taxol 10 nmol/L， BD 10 μmol/L+Taxol 20 nmol/L， BD 15 μmol/L+Taxol 30 nmol/L， BD 20 μmol/L+Taxol 40 nmol/L） were 0.63±0.04， 0.68±0.08， 0.89±0.12 and 0.84±0.05. After treated with 20 μmol/L BD， 40 nmol/L Taxol and two-drug combination， the formation of clonal colonies was decreased with different degrees of chromatin concentration and nuclear shrinkage； the rate of clone formation （24 h）， the expression of Bcl-2 （48 h）， PARP （48 h）， Caspase-3 （48 h） and JNK （4， 6 h， except for Taxol group） were decreased significantly， while the relative expression of Cleaved-caspase-3 （48 h） and p-JNK （4， 6， 12 h） were increased significantly. Those of BD+Taxol group were significantly better than those of BD group and Taxol group [except for JNK （4， 6， 12 h）， p-JNK （4 h）] （P＜0.05 or P＜0.01）. CONCLUSIONS： Both BD and Taxol can inhibit the proliferation and promote apoptosis of human pancreatic cancer Capan-2 cells， and the combination have a certain synergistic effect， which is better than any single drug. It may be associated with activating Caspase pathway and JNK phosphorylation.

OBJECTIVE： To observe the inhibitory effects and possible mechanism of new small molecular kinase inhibitors Ibr-7 [Irutinil（Ibr） derivatives] on human pancreatic cancer Capan-2 cells. METHODS： Taking Capan-2 cells as objects， CCK-8 method was used to determine the proliferation of cells after treated with 1， 2， 4， 8 μmol/L Ibr/Ibr-7 for 48 h. The survival rates of cells were calculated. Sensitization effects of 1 μmol/L Ibr/Ibr-7 on different doses of gemcitabine/paclitaxel （0.062 5， 0.125， 0.25， 0.5， 1 μmol/L） were detected. Clone formation test was used to detect the situation of cell clone formation after treated with 1， 2， 4 μmol/L Ibr/Ibr-7 for 48 h. The number of cell colony formation was recorded. Flow cytometry or JC-1 method was used to detect the apoptosis of cells after treated with 2， 4， 8 μmol/L Ibr-7 for 24 or 16 h and the changes of mitochondrial transmembrane potential； total apoptotic rate and the percentage of mitochondrial membrane potential decrease were calculated. Western blotting was used to detect the expression of related apoptotic protein （PARP， Noxa， Bcl-2， Bax， Mcl-1， Bcl-xL）. RESULTS： After treated with 1， 2， 4， 8 μmol/L Ibr/Ibr-7 for 48 h， the survival rates of cells were decreased significantly； those of Ibr-7 groups were significantly lower than those of same-dose Ibr groups； IC50 of Ibr-7 was significantly lower than that of Ibr （P＜0.05 or P＜0.01）. After combined with Ibr/Ibr-7， the survival rate of cells was significantly lower than that of same-dose gemcitabine/paclitaxel alone group， and the Ibr-7 combination group was significantly lower than same-dose Ibr combination group （P＜0.05 or P＜0.01）. After treated with 2， 4 μmol/L Ibr and 1， 2， 4 μmol/L Ibr-7 for 48 h， the number of cell clone formation was decreased significantly， while Ibr-7 groups were significantly lower than same-dose Ibr groups （P＜0.01）. After treated with different doses of Ibr-7 for 24 or 16 h， total apoptosis rate of cells （2， 4， 8 μmol/L）， the proportion of cell mitochondrial membrane potential decrease （8 μmol/L）， the relative protein expression of Noxa （2， 4， 8 μmol/L） and Bax （8 μmol/L） were increased significantly， while the protein expression of PARP （8 μmol/L）， Bcl-2 （4 μmol/L）， Mcl-1 （2， 4， 8 μmol/L） were decreased significantly； above indexes （except for relative expression of PARP and Bcl-2） of 8 μmol/L Ibr-7 group were significantly better than same-dose Ibr group （P＜0.05 or P＜0.01）. There was no statistical significance in protein expression of Bcl-xL among those groups （P＞0.05）. CONCLUSIONS： Compared with Ibr， Ibr-7 has better inhibitory and apoptotic effects on human pancreatic cancer Capan-2 cells in vitro， and has stronger chemotherapeutic drug sensitization activity， the mechanism of which may be associated with reducing mitochondrial transmembrane potential， down-regulating the protein expression of PARP， Bcl-2 and Mcl-1 and up-regulating the protein expression of Noxa and Bax.