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.

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

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

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 investigate the effects of chemokine (C-X-C motif) ligand 8 (CXCL8) gene silencing on cell proliferation and invasion of human colon cancer cells, and to investigate its relationship with phosphatidylinositol 3-kinase/protein kinase B/nuclear factor-kappa B (PI3K/Akt/NF-κB) signaling pathway. Methods: The expression levels of CXCL8 mRNA and protein in four human colon cancer HT-29, WiDr, CaCo-2 and CoLo320 cells were detected by reverse transcription-PCR and Western blotting, respectively. The four colon cancer cells were transfected with the small interfering RNA targeting CXCL8 gene (CXCL8 siRNA) by LipofectAMINE 2000, then the expression of CXCL8 protein was detected by Western blotting. The proliferation and invasion abilities of colon cancer cells after CXCL8 gene silencing were detected by WST-1 method and Transwell invasion assay, respectively. The phosphorylation levels of PI3K, Akt and NF-κB proteins in HT-29 cells with CXCL8 gene silencing were detected by Western blotting. Results: CXCL8 mRNA and protein were highly expressed in four colon cancer cell lines. After CXCL8 siRNAs were transfected into the colon cancer cells, the expression of CXCL8 protein was significantly inhibited (all P < 0.01), while the proliferation and invasion abilities of colon cancer cells were significantly decreased (all P < 0.01). CXCL8 gene silencing resulted in blockage of PI3K, Akt and NF-κB protein phosphorylation induced by CXCL8 in colon cancer HT-29 cells (all P < 0.01). Conclusion: CXCL8 gene silencing significantly inhibits the proliferation and invasion of colon carcinoma cells. It may be related to down-regulation of protein activity in PI3K/Akt/NF-κB signaling pathway.

End-stage renal disease (ESRD) can lead to various serious neurological complications, acute cerebrovascular disease and cognitive impairment are the most common ones. In recent years, a variety of new MRI techniques have been applied to elucidate the underlying neuropathological mechanisms of brain injuries in ESRD, and some progresses has been made, which is of great significance for early diagnosis and treatment of this disease. The progresses of mechanisms of brain injuries of ESRD based on MRI were reviewed in this article.

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.

Objective To identify high affinity ligand MULT1-transgenic mice,analyze its elementary phenotype,and to perform primary functional study in MULT1.Methods We identified the genotype of transgenic mice by PCR and detected transcription level of MULT1 in tails by real-time PCR.The flowcytometry and immunohisto-chemistry were used to analyze the expression of MULT1 in various tissues.Results 25 3'S-strain and 13 4'S-strain MULT1-transgenic mice were obtained and which contain high level of MULT1 transcripts.MULT1 protein is mainly expressed in intestinal intraepithelial lymphocytes (iIELs) and thymocytes but not on splenocytes.Moreover,the levels of MULT1 expression in iIELs and thymocytes are associated with age.Within iIELs,the percentage of CD8~+ Tcells decreased,while CD4~+ CD25~+ Treg cells increased,when compared with wild-type mouse.Conclu-sion We successfully produced MULT1-transgenic mice.MULT1 is possibly associated with thymic development and aging,as well as immunoregulation in mice.