Objective:To clone and express mouse fibroblast growth factor eight(FGF 8) and investigate the function of it Methods:According to the published sequence of mouse fibroblast growth factor 8, a pair of special primers was designed Then the full length cDNA of FGF 8b (a predominant isoform of FGF 8) was isolated from the total RNA of the mouse embryo by means of reverse transcription PCR and subcloned into the yeast expression vector of pYEX4T 1 in correct direction It was identified with sequencing The recombinant plasmid was transformed into yeasts The fusion protein expressed was verified by using Western blot The activity of the protein was examined by MTT and 3H TdR incorporation Results:The cDNA fragment was about 600 bps, and proved to be "b" isoform of FGF 8 exactly This fusion protein, with molecular weight about 55 kD, was highly expressed in yeast system The data of MTT and 3H TdR incorporation in NIH3T3 cells were increased obviously by the protein, and decreased by the antagonist Conclusion:Fibroblast growth factor eight was cloned and expressed.The protein was found to be capable of stimulating the proliferation of NIH3T3 cells Moreover, the recombinant protein of the extracellular fragment of FGFR1 can antagonize the response of NIH3T3 to the recombinant protein

Objective: To compare the differences in gene expression between murine intestine intraepithelial lymphocytes (iIELs) and splenic T lymphocytes, establishing the cDNA subtractive library of iIELs, analyzing the iIELs special genes. Methods: The iIELs and splenic T lymphocytes were isolated by density gradient in Percoll and nylon column respectively. cDNA subtractive library of iIELs was established by improved subtractive hybridization. The cDNA was ligated with a different adaptor. After 2 rounds of hybridization and suppression PCR amplification, the PCR products were directly inserted into the plasmid vectors. The ESTs special for iIELs were screened by reverse Northern blot. Results: Fifty ESTs with obvious difference in the cDNA subtractive library of iIELs were screened. These ESTs were squenced and submitted to Genebank. The ESTs were analyzed through BLAST, and 14 genes special for iIELs were obtained. These genes were associated with the pattern recognition of pathogen, cytokine signal transduction and proteinase inhibitors. Conclusion: iIELs are important components of intestine natural immunity, it can recognize the conservative antigens of bacteria and can be affected by cytokines and proteinase inhibitors, leading to unique cellular behavior--more resting and less activation.

16S rDNA sequences of 30 Bacillus strains originally identified as Bacillus licheniformis from China Center of Industrial Culture Collection (CICC) were determined and analyzed. The results indicated that 24 strains are affiliated to Bacillus licheniformis;3 strains are affiliated to Bacillus cereus and 1 strain is affiliated to Bacillus subitilis;the similarity levels of 16S rDNA among the rest of 2 strains and other strains of Bacillus licheniformis,range from 96.4% to 97.4%,further tests are needed to clarify their position. Also we testified that 5' terminal 500bp of 16S rDNA is available to differentiate the strains of Bacillus licheniformis、Bacillus subtilis and Bacillus cereus.

Objective To investigate the subchronic toxicity of a Chinese medicine Danhong injection in dogs . Methods Twenty-four healthy Beagle dogs (body weigh 7-9 kg, male:female=1:1) were used in this study.The test dogs were administered with Danhong injection by intravenous injection once every day for consecutive 6 days/week for thirteen weeks.The Danhong injection was used in 3 doses:494,247,and 124 mg· kg-1, repectively.The control dogs received normal saline injection in the same volume .The body weight, blood biochemistry, hematology, viscera relative weight and histopathology were determined for the overall toxicity assessment .Results The dogs administered with 494 g· kg-1 Danhong injection showed significantly reduced body weight gain , abnormal increase of urea nitrogen ( BUN ) , creatinine (CR) levels, and histopathological changes in the liver .Conclusions Danhong injection (494mg· kg-1 ) administered Intravenously has obvious toxicity on the liver and kidney function .

Objective To observe the relations of sleep structure changes and cognitive behavior abnormalities in children with idiopathic epilepsy.Methods All night polysomnographies, day attention test and Achenbach child behavior checklist were done on 64 children with idiopathic epilepsy and 20 healthy controls the requirement. Spearman correlations were made to evaluate the correlations between the parameters of sleep structure and the results of attention and cognitive behavior abnormalities.Results All children with epilepsy had longer stage Ⅰ sleep percentage and latency of rapid eye movement (REM) sleep compared with controls (all P

Apoptosis ; Cell Proliferation ; Cell Transformation, Neoplastic ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Humans ; MicroRNAs ; classification ; genetics ; metabolism ; physiology ; Neoplasms ; genetics ; metabolism ; pathology ; Signal Transduction

Pancreatic neuroendoerine tumors (PNET) is a complicated and heterogeneous group of neoplasia,and the incidence of PNET is growing rapidly recently.The current study on PNET lags behind what is known of other pancreatic tumors.Therefore,understanding of the molecular pathology is important for improving the early diagnosis,treatment and prognosis.With the development of molecular biology,there are already several potential targets proved to have application role in treatment of PNET.Here,the authors present the current knowledge about the molecular alterations of PNET.

Objective To study the relationship between the expression of NKG2D ligands and oxidative stress,and to analyze the effect of oxidative stress on the function of NK cells. Methods Tumor cells were cultured and exposed to hydrogen peroxide to develope an oxidative stress model. Then to detect the expression of NKG2D ligands in cells by Real-time PCR and Flow Cytometry. The cytotoxicity of NK cells to tumor cells was detected and compared by CCK-8 kit before and after oxidative stress. Results The expression of NKG2D ligands was induced by oxidative stress,however the NKG2D ligands induced was variable. The up-regulation of NKG2D ligands increased the cytotoxicity of NK cells efficiently,and this effect was blocked by anti-NKG2D antibody. Conclusion The expression of NKG2D ligands can be selectively induced by oxidative stress on tumor cells,and the improvement of the cytotoxicity of NK cells may enhance the immune responses accordingly.

Objective To express functional haemegglutinin(HA)protein in two different bacularvirus expression systems.Methods The whole open reading frame of A/Sichuan/1/2009(H1N1)HA was obtained by synthesis,and the HA protein were expressed in insect cells by two different bacularvius expression systems:BaculoGold system and Bac-to-Bac system. Soluble HA protein was identified by Western blot and haemegglutination test. Results The correct full length of HA gene was obtained and cloned into pAcGP67B and pFAST Bacl vectors,respectively.After 3 rounds of virus amplifyjng by re-infection of Sf9 cells,the HA protein was detected in supematant of BaculoGold system and in intracellular of Bac-to-Bac system which is much better than the former.Purified HA protein was positive not only identified by Western blot,but also detected by haemegglutinin test. Conclusion Functional HA protein was successfully expressed in two distinct bacularvirus expression systems,of which the Bac-to-Bac bacularvirus expression system is more suitable for expression of A/Sichuan/1/2009(H1N1)HA protein.

AimThe aim of this study was to establish a rodent model with similar characters of human metabolic syndrome(MS).Methods Three species mice and Wistar rats were fed with high energy chows(HEC)for 6 to 23 weeks.Animals were weighted every week.Fasting blood glucose(FBG)together with total cholesterol(TC)and low density lipoprotein-cholesterol(LDL-C)were investigated by oxidase test every two week.And fasting blood insulin(FINS)was determined by radioimmunoassay.Homeostasis model assessment of insulin resistance(HOMA-IR)was calculated as FBG×FINS/22.5.At the end of the experiment,oral glucose tolerance test(OGTT)was performed.Then animals were decapitated,and coel-fat and orchio-fat were collected and weighted to calculate the visceral fat coefficient(VFC).Results FBG,serum TC and LDL-C significantly increased(P<0.01)after 6 weeks feed of HEC in KM mice.The mice also formed abdominal obesity and insulin resistant together with impairment of glucose tolerance(P<0.05 or P<0.01).Though similar to the KM mice,C57BL/6 and BALB/c mice couldn't form abdominal obesity while the latter had increased body weight(P<0.05 or P<0.01).Wistar rats formed hyperlipidemia from 1 to 10 week and hyperglycemia from 10 to 23 week together with insulin resistance and impaired glucose tolerance(P<0.05 or P<0.01).Conclusion KM mice feed with HEC for 6 weeks could successfully establish metabolic syndrome mice model which might be suitable for drug-screening,the major characters includes the formation of abdominal obesity(increase of VFC),the increase of serum TC,LDL-C,FBG and HOMA-IR,and the decrease of OGTT.