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.

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

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 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 .

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.

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.

Adolescent ; Adult ; Anemia, Hemolytic, Autoimmune ; drug therapy ; Female ; Humans ; Immunosuppressive Agents ; therapeutic use ; Male ; Middle Aged ; Sirolimus ; therapeutic use ; Treatment Outcome ; Young Adult

Objective To investigate the effect of celecoxib on the cell cycle and cell autophagy of HepG2 cell line and its possible molecular mechanism. Methods HepG2 cells were treated with celecoxib at different concentrations (0, 50, 100, 200, 500μM), then MTT was used to detect the cell proliferation, cell cycle and apoptosis were detected by flow cytometry, the cell autophagy was observed by transmission electron microscopy, and the expressions of cyclin and autophagy protein were determined using Western blot. Results Celecoxib inhibited the proliferation of HepG2 cells in a concentration dependent manner (0, 50, 100, 200 and 500 μM) and time-dependent manner (24,48h)(P<0.05). There was no significant change in necrocytosis and apoptosis after ttreated by different concentrations of (0, 50, 100, 200 and 500 μM). The celecoxib inhibited cell cycle arrest at G1 phase, the cell rate of G1 phase increased, while the cell rate of S phase decreased in a concentration dependent manner (0, 50, 100, 200 and 500 μM) and time-dependent manner (0, 8, 16, 24h)(P<0.05). The protein expressions of cyclin D1,cyclin D3, cyclin E2, CDK2 and CDK4 significantly decreased by 500 μM celecoxib (P<0.05). The celecoxib induced autophagy in HepG2 cells, and transformed autophagy protein LC3-Ⅰto LC3-Ⅱ in a concentration dependent manner. Conclusion Celecoxib can effectively inhibit the proliferation of human malignant hepatocellular carcinoma cell line HepG2, which provides a new idea for the clinical application of celecoxib.