OBJECTIVE:To study in vitro effects of tanshinoneⅡA to reverse multiple drug resistance. METHODS:MCF-7/ADM cells and A549/DDP cells were cultured with 0 [20 mg/L doxorubicin(ADM)or cisplatin(DDP),negative control],5 mg/ml tanshinone ⅡA(combined with 20 mg/L ADM or DDP)for 24 and 48 h. Then MTT method was used to determine cell viability, and polymerase chain reaction(PCR)was adopted to detect mRNA expressions of cell cycle control protein CDC25A and cell cy-clin dependent kinase (CKD2). RESULTS:Compared to the negative control,after MCF-7/ADM cells and A549/DDP cells were cultured with tanshinone ⅡA for 24 and 48 h,cell viability was weaker,also were mRNA expressions of CDC25A and CKD2. There were statistical differences(P<0.01). CONCLUSIONS:Tanshinone ⅡA combined with ADM or DDP can inhibit the viabili-ty of cell line MCF-7/ADM and cell line A549/DDP,decrease expression of CDC25A,CKD2 mRNA in cells and reverse multiple drug resistance in malignant tumors.

Objective To develop an in vitro culture system for Cryptosporidium parvum in Madin-Darby canine kidney(MDCK) cell and observe its life cycle(from desquamate to oocyst).Methods Oocysts of C.parvum were co-cultured with MDCK cells in vitro.Culture condition was optimized and the life cycle of C.parvum investigated.Results The optimal culture conditions for C.parvum in MDCK cells were 2.0?105 cells cultured for 12 h, and infected by 1.0?105 oocysts in the Dulbecco′s Modified Eagle Medium with 5% FBS.Following 72 h co-culture, desquamate, sporozoites, trophozoites, meronts, microgametocytes, macrogametocytes, zygote, thin-wall oocyst, and thick-wall oocyst appeared orderly.Between the 60th and 72th hour, many oocysts emerged.Inoculated by the C.parvum-infected cell culture supernatant at the 48th hour, the immunosuppressed mice became infected.Conclusion The culture system provides a model for propagation of the parasites and demonstrates a complete in vitro life cycle of C.parvum.

Objective To explore an applicable method for isolation and purification of Cryptosporidium parvum oocysts with high purity, recovery and vigor from mouse feces. Methods Four techniques were used for isolating and purifying C.parvum oocysts from mouse feces: modified saturated saline flotation, percoll gradient centrifugation, CsCl gradient centrifugation and the classical discontinuous sucrose gradient centrifugation. Oocysts received from the methods were used respectively to infect in vitro bovine fallopian tube epithelial cells (BFTE) and the development of the oocysts was examined under microscope after 48 h and 72 h cultivation. Results The number of oocysts received by the classical discontinuous sucrose gradient centrifugation [(2.86?0.08)?107] was significantly higher than that of percoll gradient centrifugation [(1.52?0.08)?107] (P0.05). Oocysts received from CsCl gradient centrifugation showed higher purity than those by discontinuous sucrose gradient centrifugation. Conclusion In comparison to the classical discontinuous sucrose gradient centrifugation, operation of the modified saturated saline flotation is easier and faster, and the purity of oocysts isolated by CsCl gradient centrifugation is higher.

Purpose:To study the characteristics of biological distribution in vivo and therapeutic efficacy of 125 I-anti-alpha-fetoprotein(AFP) antibody when administrated by the hepatic artery , and discussion of multi-modal therapy by transcatheter arterial chemoembolization(TACE) and immunization therapy. Methods:21 patients with moderately and advanced primary liver cancer (PLC) were treated by 125 I-anti -alpha-fetoprotein which was administered via hepatic artery, together with TACE and CD3AK cell by intravenous infusion to,detect the pharmacokinetic parameters and metabolism for 125 I-anti-AFP in vivo, and observe the efficacy of multi-modal treatment. Results:The radioactivity half-life time of ?phase (T 1/2 ?) and ? phase ( T 1/2 ?) for 125 I-anti-AFP antibodies was 1.85?1.79 and 156.46?65 hr,respectively; half-excretion time from urine was 94 hr, radiation intensity measured at body surface of organ suggested that the accumulated radioactivity was stronger and longer in the liver was than other organ.TPECT for tumour:liver ratio was 2.1?0.6. The efficacy for the treated group and the control group was respectively 61.9%(13/21) and 25.0%(5/20),the one-year cumulative survival rate of the treated group and the control group was also 61.9% and 25.0%.Conclusions: 125 I-anti-AFP via hepatic artery was able to concentrate electively in the tumour ,thereby bringing about a continuous internal irradiation for the tumour.This combined treatment is an effective method for PLC.

Objective To study the change of DNA polymerase beta and XRCC1's expression during malignant celI differentiation.Methods The Eca-109 cells were divided inm 2 groups:differentiation group which cultured with 8-Br-cAMP and control group.The 2 groups cells were cultured 48h simultaneously.The immunocytochemistry was performed to detect the expression of DNA polymerase beta and XRCC1.Results Compared with control group,the expression of DNA polymerase beta and XRCC1 was decreased simultaneously(P＜0.05).Conclusion The differentiation agent can down-regulate the expression of DNA polymerase beta and XRCC1,suggesting that overexpressed DNA polymerase beta and XRCC1 maybe result in mutator phenotype.

Objective: To explore the effect of different selenium sources on the function of humoral immunity and antioxidant capacity of rabbits. Method: Thirty-five rabbits were randomly divided into seven groups and vaccinated with rabbit haemorrhagic disease (RHD) dead vaccine. At the same time, rabbits were injected respectively with sodium selenite (0.1 mg/kg bw and 0.3 mg/kg bw), Kappa-selenocanageenan (0.1 mg/kgbw and 0.3 mg/kg bw), DL-selenomethionine (0.1 mg/kg bw and 0.3 mg/kg bw) and physiological saline as control. Antibody against RHD, activity of GSH-Px and content of MDA in rabbit serum were detected on 0, 10, 20, 30d after inoculation. Results: Sodium selenite (0.1 mg/kg bw), Kappa- selenocanageenan (0.3 mg/kg), and DL-selenomethionine (0.3mg/kg bw) could significantly increase the level of RHD antibody. Sodium selenite (0.3 mg/kg bw) and Kappa-selenocanageenan (0.3mg/kg bw) improved the activity of GSH-Px. All selenium groups could decrease serum MDA, but Kappa-selenocanageenan (0.3 mg/kg bw) showed the best effect. Conclusion: Kappa-selenocanageenan (0.3 mg/kg bw) was better than the lower dosage and other selenium sources in the effects on the function in humoral immunity and antioxidant capacity of rabbits.

Fecal samples of 2,056 dairy cattle from 14 farms were collected in three geographical regions of China and stained using a modified acid-fast staining technique to identify Cryptosporidium oocysts. A total of 387 (18.82%) positive samples were identified and further analyzed by polymerase chain reaction (PCR) using primers designed to amplify DNA fragments from the small subunit ribosomal RNA. The PCR products were sequenced and the sequences were deposited in the GenBank database under accession numbers EU369377-84 and GU070730-33. Phylogenetic analysis was performed and a distances matrix generated from these sequences confirmed the existence of Cryptosporidium (C.) parvum 'mouse' genotype, C. bovis, C. andersoni, C. hominis, and C. serpentis in cattle. These results represent the first report on the prevalence and genetic identification of Cryptosporidium species, and may contribute to a better understanding of the epidemiology of Cryptosporidium in cattle in China.
Animals ; Base Sequence ; Cattle ; Cattle Diseases/epidemiology/*parasitology ; Chi-Square Distribution ; China/epidemiology ; Cryptosporidiosis/epidemiology/parasitology/*veterinary ; Cryptosporidium/genetics/*isolation & purification ; DNA, Protozoan/chemistry/genetics ; Feces/parasitology ; Female ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction/veterinary ; Prevalence ; RNA, Ribosomal, 18S/chemistry/genetics ; Sequence Alignment ; Sequence Analysis, DNA

The purpose of the study is to construct recombinant goat pox virus (GPV) expressing Peste des petits ruminants virus (PPRV) H protein, and to evaluate the immunization effect. Recombinant GPV containing PPRV H gene (rGPV-PPRV-H) was selected and purified by gpt and eGFP utilizing plaque purification, and the final selected recombinant GPV was proved to be purified by PCR. Immunofluorescence and Western blotting showed that the recombinant virus could express H protein of PPRV while infecting lamb testis cells. Six goats were immunized with 2 x 10(6) PFU rGPV-PPRV-H through intradermal injection, and were immunized for the second time at 28 days with the same dose recombinant virus after first immunization. Serum was collected after immunization, and was analyzed for the neutralization antibodies. 21 days after first immunization, the neutralization antibodies of GPV were 40, 80, > or = 80, > or = 80, 40, > or = 80 in turn, and neutralization antibodies of PPRV were 80, 80, 80, 80, 40, 40, 10 in turn; 14 days after second immunization, the neutralization antibodies of GPV were all > or = 80, and the neutralization antibodies of PPRV were > 80, 80, > 80, 80, 80 and 40 in turn. This study established a foundation for the industrialization of the PPRV recombinant GPV vaccine.
Animals ; Capripoxvirus ; genetics ; immunology ; Goat Diseases ; immunology ; prevention & control ; virology ; Goats ; Hemagglutinins, Viral ; genetics ; immunology ; metabolism ; Peste-des-Petits-Ruminants ; immunology ; prevention & control ; Peste-des-petits-ruminants virus ; genetics ; immunology ; Recombinant Proteins ; genetics ; immunology ; metabolism ; Vaccines, Combined ; immunology ; Vaccines, Synthetic ; immunology ; Viral Vaccines ; immunology