Different subspecies or strains of the same species produce varied clinical manifestations.The clarifica-tion of parasite taxonomy is useful for the researches of their biology, epidemiology and control.DNA molecular markers have the advantages of high polymorphism, non-pleiotropy, and clear identifying alleles, etc.This paper summarizes the first generation(restriction fragment length polymorphism, random amplified polymorphic DNA), second generation(sim-ple sequence repeat-anchored PCR, inter-simple sequence repeat) and third generation(single nucleotide polymorphism) molecular marker techniques, and their application in taxonomic identification of parasites.

Objective To observe the efficacy and safety of the cocktail therapy to treat patients with Parkinson's disease with dementia ( PDD) . Methods Sixty patients with PDD were randomly divided into treatment group(30 cases)and control group(30 cases). The dose 10 mg of donepezil hydrochloride was taken 1 time a day in the control group;on the basis,the dose 0. 2 g of butylphthalide soft capsules was taken pre-dinner, 3 times a day,the dose 0. 8 g of oxiracetam was taken 3 times a day and 2 Ginkgo Biloba Leaves Extract tablets was taken 3 times a day in the treatment group,period of treatment was 6 months. The curative effect was measured by using Montreal cognitive assessment scale (MoCA),Blessed-Roth dementia table,severity of clinical dementia rating scale (WMS) rating, respectively before treatment and on the 3th, 6th month after treatment,and monitored the adverse reactions. Results There were no statistically significant differences on the scores of MoCA, WMS and Blessed-Roth dementia table between treatment group and control group before treatment ( all P>0. 05 ) . The scores of MoCA, WMS in the treatment group and control group on 6 months after treatment were significantly higher than those before treatment and 3 months after treatment, and the scores of Blessed-Roth dementia table in the treatment group and control group on 6 months after treatment were significantly lower than those before treatment and 3 months after treatment ( P<0. 05-0. 01). Compared with control group,the scores of MoCA, WMS increased obviously and the scores of Blessed-Roth dementia table decreased significantly in the treatment group on 6 months after treatment (P<0. 05-0. 01). There was no statistically significant difference on the rates of abnormal liver function after treatment betweenin the treatment group (16. 67%) and the control group (13. 33%) (P>0. 05). Conclusion The curative effect of cocktail therapy is better than traditional remedies for treating patients with PDD, and it is high safety.

Objective To study the expression of VEGF and microvascular density (MVD) in pancreatic carcinoma and their prognostic value. Methods The protein expression of VEGF and MVD were determined with immunohistochemistry in 48 cases of pancreatic ductal carcinomas, which were surgically resected and pathologically confirmed. Results In 64.6% of pancreatic carcinomas the expression of VEGF was positive, and it was significantly higher than that of normal pancreatic tissues (P

Objective To study the damage effect of oxidized low density lipoprotein (OX-LDL)on the endothelial cells and its influence in the expressions of Toll like receptor 4 (TLR-4)and epithelial cell adhesion molecule (EpCAM).Methods The human endothelial cells ECV-304 were cultured in vitro and treated by different concentrations of OX-LDL for 24 h,and the cell vitality was measured by Taipan blue rejection test.The reactive oxygen species (ROS)level,mitochondrial membrane potential (MMP),cell cycle and apoptosis were detected by flow cytometry (FCM).The expression of TLR-4 and EpCAM were also analyzed by FCM.Results Compared with control group,the ROS levels in ECV-304 cells in 25,50,100,and 200 mg·L-1 OX-LDL were increased, but the cell vitalities and MMP were decreased (P < 0.05).Compared with control group,the percentages of S phase cells in 25 and 50 mg·L-1 OX-LDL groups were increased,the G0 G1 phase cells were slightly reduced,and the SubG1 phase cells (apoptotic cells)were increased with the increasing of OX-LDL concentration.Compared with control group,the TLR-4 and EpCAM expression levels in ECV-304 cells in 50 mg·L-1 OX-LDL group at 24 h were significantly increased (P < 0.05 ). Conclusion OX-LDL could increase the ROS level and induce the apoptosis in vascular endothelial cells,while increase the cell vitality and MMP.During the damage process,the expresion levels of TLR-4 and EpCAM are up-regulated.

Objective To observe the protective effects of FTY720 on the Con A-induced mouse hepatic fibrosis injury,and to find the possible mechanisms of protective effects.Methods The pathologic models of hepatic fibrosis injury in the mice caused by Con A were set up.Forty mice were randomly divided into control group, model group,high dose of FTY720 (4 mg·kg-1 )group and low dose of FTY720 (1 mg·kg-1 )dose group (n=10).The serum alanine aminotransferase (ALT)and asparate aminotransferase (AST)activities,hepatic index and pathological changes of hepatic tissue were detected .Results Compared with model group,the serum ALT and AST activities in low and high doses of FTY720 groups were decreased significantly (P < 0.05 or P < 0.01).The optical microscope results showed that there were inflammatory cells and hepatocellular necrosis in model group. The masson staining results showed that there were surrounding fiber bundle and hepatic lobule fusion in model group;compared with model group,the damage degree in low and high doses of FTY720 groups was reduced.The protective effects of FTY720 on hepatic injury showed linear relation to the drug dose.Conclusion FTY720 could decrease the levels of ALT/AST,thus FTY720 alleviate hepatic damage degree and delay the process of hepatic fibrosis.The protective effects of FTY720 on hepatic injury in experimental hepatic fibrosis mice may be related to the mechanisms mentioned above.

Objective: To investigate the effects of culture medium osmolarity on the proliferation of human hepatoma cell line HHCC. Methods:Cell proliferation was analyzed by MTT assay and cell cycle analysis was carried out by flow cytometry. Results:Hyperosmolarity decreased significantly the absorbance (A) of HHCC cells and cell number, and hyposmolarity have counter effects. Compared with control, hyperosmolarity blocked cell cycle progression at the G 1 phase, while hyposmolarity accelerated cells into S phase from G 1 phase, and the effect of hyposmolarity was weakened by chloride channel blocker NFA. Conclusion:Culture medium osmolarity influenced the proliferation of HHCC cells by mediating cell cycle.

Objective To study the apoptosis of K562 cells induced by a new immunosuppressive agent FTY720 and its mechanism,and to provide experimental basis for the treatment of leukemia in clinic.Methods The K562 cells were cultured in vitro and divided into blank control group and FTY720 treatment group.The K562 cells in FTY720 treatment group were treated with 6μmol·L-1 FTY720 for 3,6,and 12 h,or treated with different concentrations of FTY720 (2,4,6,8μmol·L-1)for 24 h.The apoptosis,level of reactive oxygen species (ROS),mitochondrial membrane potential(MMP)and cell cycle were measured by flow cytometry.The inhibitory rate of proliferation of K562 cells after treated with FTY720 was detected by WST-1 reducting assay.Results The results of flow cytometry showed that the percentages of apoptotic cells were increased after treated with 6μmol·L-1 FTY720 for 3,6,and 12 h with the prolongation of time compared with blank control group(P<0.01).The percentages of apoptotic cells were also increased after treated with different concentrations of FTY720 for 24 h compared with blank control group(P<0.01).Compared with blank control group,the ROS levels were increased with the increasing of FTY720 concentration,while the MMP was decreased(P<0.01).Compared with blank control group,the percentage of cells at G0/G1 phase was increased,while those at S and G2/M phases were decreased with the increasing of FTY720 concentration (P<0.05).The WST-1 reduction assay results indicated that the inhibitory rates of proliferation of K562 cells after treated with 2,4,6,and 8μmol·L-1 FTY720 for 72 h were (24.0±4.1)%,(46.4±3.9)%,(67.0±4.8)%,and (88.2±5.6)%,respectively,compared with blank control group.The concentration of FTY720 to result the inhibitory rate of 50% (IC50 )on K562 cells was 5.5μmol·L-1 .Conclusion FTY720 could inhibit the proliferation of K562 cells by blocking cell cycle and inducing apoptosis through provoking ROS.

AIM: To investigate the changes of the redox status and the antioxidative capability in the tissue of malignant tumors. METHODS: The carcinoma tissues collected from 42 patients with primary cancer in digestive tract (13 cases of esophageal cancer, 14 cases of gastric cancer and 15 cases of colorectal cancer),the corresponding paratumor mucosa tissues were taken as the control samples. The content of oxidized and reduced glutathion (GSSG and GSH), oxidized and reduced coenzyme II (NADP+ and NADPH) were measured, the GSH/GSSG, NADPH/NADP+ ratios, and the GSH/GSSG, NADPH/NADP+ redox potentials were calculated according to Nernst formula. RESULTS: The levels of GSH and NADPH in cancer tissues were significantly higher than those in corresponding paratumor tissues (P0.05). CONCLUSIONS: The significant increase in GSH and NADPH contents in cancer tissues indicates a notable enhancement of its antioxidative capability compared with the corresponding paratumor tissues. Based on this changes, the redox potential in the cancer tissues has only slightly reductive shift, which may suggest an apparent oxidative stress existed in the cancer tissues.

To establish an HPLC method to determine osimertinib mesylate,Agilent ZORBAX Eclipse Plus C18 column (4.6 mm × 250 mm,5 μm) was used with a mobile phase consisting of methanol-buffer solution (20 mmoL/L NaH2PO4,pH 3.0 adjusted with 85％ H3PO4) (50 ∶ 50) at the flow rate of 1.0 mL/min.The detection wavelength was 210 nm,and the column temperature was kept at 35 ℃.The calibration curve was liner over the range from 50％ to 150％ of determination concentration (0.201 1-0.603 2 mg/mL,r =0.999 9).The limit of quantitation (LOQ) and limit of detection (LOD) were 0.32 μg/mL and 0.08 μg/mL,respectively.The contents of osimertinib mesylate in samples were 100.1％,99.5％ and 99.7％.Good chromatographic separation of osimertinib mesylate and its related substances,including synthetic impurities and degradation products,were obtained.The established HPLC method is specific,accurate,simple and durable,and could be used for the determination of osimertinib mesylate.