Objective To investigate the dynamic changes of IL-2, IFN-γ, TNF-α, IL-4, IL-5 and IL-10 in mice immunized with Eg14-3-3 and then challenged by Echinococcus granulosus protoscoleces.Methods ICR mice were subcutaneously immunized every two weeks for altogether three times with rEg14-3-3, followed by the challenge with Echinococcus granulosus protoscoleces intraperitoneally four weeks after the third immunization.The PBS control group mice were operated in the same way.We collected mouse serum from tail vessel at different time points after immunization and challenge.The serum levels of IL-2, IFN-γ, TNF-α, IL-4, IL-5 and IL-10 were examined with enzyme-linked immunosorbent assay.Results The levels of the six cytokines were higher after immunization and challenge infection in rEg14-3-3 group than in PBS control group.The immunized mice generated a great deal of Th1 cells, namely, IL-2, IFN-γ and TNF-α, after immunization and showed a peak at week 10 (acute infection phase), then the level decreased rapidly at week 30 (chronic infection phase), and maintained a high level for a long time.In contrast, Th2 cells like IL-4, IL-5 and IL-10 kept a low level until week 18, peaked at week 30, and then decreased gradually but maintained a relatively high level for a long time.For PBS control group, IL-2, IFN-γ and TNF-α did not increase obviously before infection, but increased rapidly after challenge infection and peaked at week 18;then the level decreased gradually and maintained a high level for a long time.On the other hand, IL-4, IL-5 and IL-10 kept a low level, but increased gradually after challenge infection.IL-4 peaked at week 18 while IL-5 and IL-10 peaked at week 30, and then they all decreased slowly but maintained a relatively high level for a long time.Conclusion Eg14-3-3 can induce significant antibody response by Th2 cells and cell-mediated immunity response by Th1 cells.Both Th1 and Th2 cells participate in the anti-echinococcusis protective immunity.

Splenic radiofrequency ablation (RFA) is a safe, effective and minimally-invasive approach for the management of hypersplenism due to portal hypertension. After RFA, the remnant volume of the spleen will be decreased, the hypersplenism can be corrected, and the hepatic artery blood flow can be significantly increased with resultant marked improvement of liver function; in addition, hypertrophy and regeneration of the liver can be induced. However, many factors can affect the therapeutic results of RFA,therefore, further studies are necessary to clarify the underlying mechanisms.

Objective To analyze the differentially expressed proteomics on T lymphocyte in mice immunized with 14‐3‐3 recom‐bined vaccine and injected with phosphate buffer solution (PBS) .Methods Mice immunized with 14‐3‐3 recombined vaccine were separated in the experiment group and those injected with PBS were selected in the control group .Two weeks after immunization with 14‐3‐3 recombined vaccine ,total protein of T lymphocyte isolated from the two groups was collected and separated by using the two dimensional electrophoresis(2‐DE) .The differentially expressed proteomics were compared .Results The 2‐DE map shown 11 differentially expressed protein plots ,including 7 protein plots spotted in the 2‐DE map of T lymphocyte from 14‐3‐3 recombined vaccine immunized‐mice and 4 protein plots spotted in the 2‐DE map of T lymphocyte from PBS injected‐mice .Conclusion There are differentially expressed proteomics of T lymphocyte in PBS injected‐mice and 14‐3‐3 recombined vaccine immunized‐mice ,which could contribute to clarifing the mechanism on protective immunity of 14‐3‐3 recombined vaccine and optimizing vaccine strategy for improving vaccine effect .

Objective To screen and clone the genes related to stilbene glucosides biosynthesis in Polygonum multiflorum Thunb. Methods The differentially expressed genes in the root, stem and leaf of Polygonum multiflorum which have different contents of stilbene glucosides were screened by differential display reverse transcription polymerase chain reaction (DDRT-PCR). After pMD19-T carrier was inserted into the obtained differential genes for sequencing and comparison, the gene function was analyzed. Results Fifty-one differentially expressed cDNA fragments were found. Of them, 9 were used for the identification by semi-quantitative PCR. The identification results presented 3 positive fragments, one fragment was specifically expressed in the stem and leaf of Polygon-um multiflorum Thunb., sharing high homology with glycine dehydrogenase, and 2 were specifically expressed in the root of Polygonum multiflorum Thunb., having high homology with enoyl-CoA hydratase and aminopeptidase N, respectively. Conclusion Three homologous gene sequences obtained through DDRT-PCR provide a basis for the further study of biosynthetic pathway of stilbene glucoside from Polygonum multiflorum Thunb..

OBJECTIVETo establish and optimize the proteomic analysis of protoscoleces-specific antigens from Echinococcus granulosus. To provide a foundation for identifying specific antigens in the soluble proteins of E. granulosus protoscoleces for further research.

METHODSBrood capsules were collected aseptically from fertile E. granulosus cysts from the livers of an infected patient. The fertile E. granulosus cysts were fractured, and protoscoleces were collected by centrifugation. The soluble proteins of protoscoleces were acquired using the 2D Quant kit according to the manufacturer's instructions. We employed two-dimensional electrophoresis (2-DE) combined with immunoblot assay (Western blot) to analyze the soluble components of E. granulosus protoscoleces antigens. The 2-DE and immunoblot maps obtained were analyzed with PDQuest 8.0 image analysis software.

RESULTSAbout 233 soluble protein spots were identified with Coomassie-stained gels. Most of the proteins had a molecular weight of 16,000 Da to 117,000 Da, and an isoelectric point value of 3.0 to 10.0. 2-DE immunoblot was conducted and 57 specific antigen spots were observed, among which 23 spots were identified.

CONCLUSION2-DE combined with Western blot is the key to successful proteomic analysis and presents a new possibility for searching the specific E. granulosus protoscoleces antigens.

Animals ; Antigens, Helminth ; chemistry ; metabolism ; Blotting, Western ; methods ; Echinococcus granulosus ; classification ; metabolism ; Electrophoresis, Gel, Two-Dimensional ; Gene Expression Regulation ; Helminth Proteins ; isolation & purification ; Proteomics ; methods

Adult ; Aged ; Electrocardiography ; Humans ; Male ; Middle Aged ; Radiography, Thoracic ; Silicosis ; complications ; diagnostic imaging

Animal models of pancreatic cancer are important for the experiments of pancreatic cancer research.An ideal an- imal model of pancreatic cancer provides effective tool for exploring the tumorigenesis of pancreatic cancer.This paper summari- zes the methods for establishing mouse model of pancreatic ductal adenocarcinoma and discusses interpretd the advantages and disadvantages of different models.

Air Pollutants, Occupational ; analysis ; Cyanamide ; analysis ; Spectrophotometry ; Workplace

OBJECTIVE:To establish a HPLC method for simultaneous determination of chlorogenic acid and cefazolin in rat plasma.METHODS: Chromatographic determination was performed on spherisorb C18 column with methyl alcohol- 0. 1mol/L potassium ethydrogen phosphate(30: 70v/v, pH 3.0)as the mobile phase and a flow rate of 1.2ml/min.Detection wavelengthes were UV 275nm for chlorogenic acid and 326nm for cefazolin.The concentrations of chlorogenic acid and cefazolin in rat plasma were detected.RESULTS: The calibration curves were linear with the correlation coefficients of 0.997 and 0.998, repectively.The RSDs for the between- day and within- day were both lower than 4% for chlorogenic acid and 6% for cefazolin, respective.The mean recoveries were 93.30%, 93.35% and 96.40% for chlorogenic acid, and 93.41%, 96.60% and 92.36% for cefazolin at large, middle and low dosages, respectively.The concentrations of cefazolin in rat plasma were decreased in coadministration with chlorogenic acid.CONCLUSION: The study provides a simple method for simultaneous determination of the plasma levels of chlorogenic acid and cefazolin.

Objective To investigate the effects of exogenous nitric oxide (NO) on gaseous transmitter molecule of the hippocampus in rats with global cerebral ischemic-reperfusion(I-R) injury.Methods Twenty-four Wistar male rates were randomly divided into 4 groups of sham operation (group S),I-R control(group I-R),I-R plus sodium nitroproside(SNP) 2 mg/kg(LSNP) or plus SNP 4 mg/kg(group HSNP) with 6 rats each.Global cerebral I-R model was established by 4-vessel occlusion.SNP was injected at 30 min before clamping the bilateral carotid arteries in groups of LSNP and HSNP.The carotid arteries were clasped for 20 min.The rats were killed after cerebral reperfusion for 6 h.The concentrations of hydrogen sulfide(H_2S),NO and CO,the activity of cystathionine beta synthase(CBS),HO and inducible nitric oxide synthase(iNOS),as well as the expression levels of CBS mRNA,iNOS mRNA and hemeoxygenase-1 (HO-1) mRNA in the hippocampus were measured.Results Compared to group S,the concentrations of H_2S,NO and CO,the activities of CBS,HO and iNOS and the expressions of CBS mRNA,iNOS mRNA and HO-1 mRNA in group I-R were increased(P<0.05 or P<0.01) ,which were all higher in groups of LSNP and HSNP than those in group I-R(P<0.05 or P<0.01).Conclusion Exogenous NO can induce the expressions of CBS mRNA and HO-1 mRNA,activate CBS and HO in rats with global cerebral I-R injury,indicating that NO has a modulation effect on the CBS/ H_2S and iNOS/NO systemes.