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 .

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 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.

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..

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.

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

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

Objective:To investigate the effects of LPS on the IL-6 expression in osteoblasts.Methods:Rat calvarial osteoblasts were primarily cultured. LPS from Escherichia coli with different concentration were administrated. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to identify the IL-6 mRNA expression of the cells. The intensities of electrophoresis bands were measured. Enzyme-linked immunosorbent assay (ELISA) was used to detect IL-6 protein production. The data were analyzed with SPSS 10.0 software package.Results:E. coli LPS with concentrations of 1?10-5 g/L-1?10-3 g/L induced IL-6 mRNA and IL-6 protein expression in osteoblasts. Statistically significant differences were detected at concentrations 1?10-4,1?10-3 g/L E.coli LPS.Conclusion:LPS can increase the expression of IL-6 in osteoblasts in the gene and protein levels. IL-6 may play an important role in the alveolar bone metabolism.

Objective To investigate the expression and significance of Toll-like receptor 4(TLR4)in the uvea of endotoxin in- duced uveitis.Design Experimental study.Participants 12 Wistar rats were divided into two groups at random,a group(n=6)of endo- toxin induced uveitis(EIU)and a control group(n=6).Methods Cholera vibrio was injected into hind footpad and abdominal cavity in the EIU group.In the control group,cholera vibrio was replaced by PBS.Anterior segment of the rats eyes were observed by slit lamp microscope at 4h,10h,16h,24h after injection.Immunohistochemical staining was performed using polyclonal antibodies to TLR4 on the frozen sections of the eyes and uveal wholemounts at 24 hours after LPS injection.Main Outcome Measures The inflammation of anterior segment and the number of TLR4 positive cells in uveal wholemounts.Results The inflammation began at 4h after LPS injec- tion.Cellulose exudates were observed at 16h after injection.The positive-stained protein located in cellular membrane or cytoplasm. The number of TLR4 positive cells in EIU group is evidently higher than that in the control group.TLR4 positive cells were hardly de- tected in the frozen sections of eyes.Conclusion TLR4 expression increases in the EIU,which suggests that TLR4 probably participates in the formation and development of acute anterior uveitis.(Ophthalmol CHN,2008,17:48-51)