Objective:To analyse the nucleotide polymorphism of dentin sialoprotein(DSP) coden region in Chinese people. Methods:The DSP segments were amplified by PCR;single-stranded conformation polymorphism(SSCP) and DNA sequencing were employed to detect the nucleotide polymorphisms in DSP coden region. Results:Three single nucleotide polymorphisms(SNP) were found in DSP coden region,two were same sense SNPs resulting in no change of amino acid product,and one was missense SNP resulting in change of asparagine and aspartic acid. Conclusion:There are some SNPs existing in DSP coden region in Chinese people.

Objective:To express human dentin sialoprotein (hDSP) gene in COS-7 cells. Methods:hDSP gene was subcloned into mammalian expression vector pcDNA3. The recombined plasmids were transfected into COS-7 cells using lipofectamune PLUS TM kit for transient expression. Western blot analysis and immunohistochemical staining were used to examine the gene products. Results:The constructed vectors were confirmed by digestion with restriction enzyme. An immuno-reaction positive band with relative molecular mass of 60 000 was found by Western blot analysis in culture supernatant and cytoplasms of COS-7 cells. Immunohistochemical staining showed strong positive particles in the cytoplasms. Conclution:hDSP gene can be expressed in COS-7 cells.

Objective To investigate the relationship between the serum ghrelin level and β-cell function after treatment of patients with newly diagnosed type 2 diabetes mellitus.Methods 34 patients with newly diagnosed type 2 diabetes mellitus were divided into three groups of pre-treatment(DM0),two weeks after treatment(DM1)and six months after treatment(DM2).Intravenous glucose tolerance tests(IVGTT)were performed and blood glucose,HbA1C,ghrelin,insulin were measured during the study.Insulin sensitivity was assessed by homeostasis model assessment(HOMA).Results(1)Compared with DM0 group,beta-cell function and the mean AUC of insulin were dramatically improved in groups of DM1 and DM2(all P<0.05).(2)Compared with DM1 and DM2,ghrelin levels and the mean AUC of ghrelin in DM0 groups were lower(all P<0.05).(3)Only positive correlation between ghrelin and HOMA-β(r=0.446,P<0.05),and negative correlations of ghrelin with glucose(r=-0.274,P<0.05),insulin(r=-0.35,P<0.05)and BMI(r=-0.276,P<0.05)were found after six months of treatment.Conclusions Ghrelin might play an important role in the pathogenesis and progress of type 2 diabetes mellitus.

Objective:To construct a suppression subtractive library of suppression-related genes from c serotype Streptococcus mutans (S.mutans). Methods:After being isolated from virulent and avirulent strain of S. mutans respectively, the intact and high-pure genomic DNAs were digested with restriction enzyme AluⅠ. The digested DNA of the avirulent strain ligated with an adaptor was used as tester DNA, and that of the virulent strain as driver DNA. Then the suppression subtractive hybridization(SSH) was carried out, the efficiency of ligation and subtraction were detected respectively. The subtracted fragments were inserted into vector pCR2.1 using T/A cloning kit and transformed into E. coli TOP10F' competent cells. The white colonies were selected to construct the suppression subtractive library. Results: Through electrophoresis of AluⅠ-digested DNAs, a smear ranged from 0.1 to 2.0 kb was observed. The ligation efficiency of tester DNA with adaptor was at least higher than 25 percent. The subtraction efficiency confirmed the success in enrichment of differential genes between virulent and avirulent strain of S. mutans. In the subtracted group, the appearance time of the 23S rRNA gene in both tester and driver DNA was later than that in the unsubtracted group by twelve cycles. It suggested that suppression subtractive hybridization happened indeed. Then the subtracted fragments were cloned and the suppression-related gene library between virulent and avirulent strain of S. mutans was constructed. Conclusions:The suppression subtracted library of suppression-related genes has been constructed.

Objective:To explore avirulent strain-specific new genes or new functions of already known genes from Streptococcus mutans of serotype c.Methods:Twenty-six DNA fragments unique to avirulent strain of Streptococcus mutans were sequenced.The sequences of these presumptive avirulence DNA fragments were subjected to homology search through BLASTn and BLASTx in public database,and their putative biological functions were analyzed.Results:The size of the DNA fragments ranged from 113 bp to 776 bp.The average G+C content was 38.27%,similar to that of the gene-codingsequences in Streptococcus mutans strain UA159 whose genome sequences were just completed.Seven clones were picked repeatedly.Of the nineteen DNA fragments,eight potentially represented new DNA fragments were registered in GenBank.The remaining DNA fragments showed high homology to known genes of Streptococcus mutans strain UA159.Conclusions: The gene analysis and identification supply the groundwork for further study of gene functions of the avirulent strain of Streptococcus mutans serotype c.

AIM: To investigate the physiological function of the novel serine protease SNC19 protein and its possible role in cancer invasion and metastasis. METHODS: Monoclonal antibodies directly against SNC19 extracellular domain was prepared. The protein and SNC19 mRNA expression were determined in different kinds of cell lines respectively by Western blot and Northern blot analysis. Cellular migration and adhesion abilities were assayed by monoclonal antibody blocking method. RESULTS: Western blot analysis showed there were two bands of SNC19 protein in BCAP37, COLO205, SW480 cells at about 120 kD and 60 kD while only one band in SW620 cells at 60 kD; Northern blots showed a approximate 3.4-kilobase fragment appearing in most epithelial-derived cell lines with this only form and high levels but no detection was obtained in OV, TCA8113, KB and SGC7901 cells. In antibody blocking experiments, the migration of SW480 cells was significantly inhibited compared with the control and the abilities of (SW480/SW480), SW480/NIH3T3 adhesion increased at the beginning of the experiments, but the difference reduced (along) with the time passed.CONCLUSION: SNC19 protein is closely related with cellular homogeneous and heterogeneous adhesion as well as cellular motility. As a novel serine protease, it may participate both in physiological and pathological processes, such as cell migration, tissue remodeling and cancer invasion and metastasis.

Objective To investigate the relationship between inducible nitric oxide synthase(iNOS)mRNA expression of pancreatic tissues and intestinal permeability,and the effect of salviae miltiorrhizae on the change in gut permeability in rats with severe acute pancreatitis(SAP).Methods All rats were randomly divided into three groups:Sham operation(group C,n=15),SAP group(group SAP,n=15),and salviae miltiorrhizae therapeutic group(group T,n=15).At 24h After operation,the iNOSmRNA expressions of pancreas were determined by in situ hybridization(ISH),and blood amylase(AMY),nitric oxide(NO) and lipopolysaccharide(LPS) were examined.The histopathologic changes of pancreas and ileum were observed.Intestinal permeability was measured by means of albumin clearance(AC)of 125I-labeled rat serum albumin.Results The iNOSmRNA expressions of pancreas in group SAP were markedly increased in comparison with group T(P

In recent years, preoperative therapy has become the standard procedure to improve radical resection rate and local control for advanced rectal cancer. Tumor responses to chemoradiotherapy, however, vary considerably, thus increasing the demand for both functional and morphologic radiologic evaluation of response to chemoradiotherapy to distinguish responders from nonresponders. MR imaging is considered the most accurate tool for the primary staging of tumor extent, and can be used to evaluate the efficacy of chemoradiotherapy. Functional imaging modalities including DW-MRI and PET-CT have shown promising prospect in the early evaluation of the response of rectal cancer to preoperative chemoradiotherapy. However, wide clinical application will take some time.
Chemoradiotherapy ; Humans ; Magnetic Resonance Imaging ; Neoadjuvant Therapy ; Rectal Neoplasms ; diagnosis ; drug therapy ; radiotherapy ; Tomography, X-Ray Computed ; Treatment Outcome

Objective To investigate the relationship between the serum ghrelin level and ?-cell function after treatment of patients with newly diagnosed type 2 diabetes mellitus.Methods 34 patients with newly diagnosed type 2 diabetes mellitus were divided into three groups of pre-treatment(DM0),two weeks after treatment(DM1)and six months after treatment(DM2).Intravenous glucose tolerance tests(IVGTT)were performed and blood glucose,HbA1C,ghrelin,insulin were measured during the study.Insulin sensitivity was assessed by homeostasis model assessment(HOMA).Results(1)Compared with DM0 group,beta-cell function and the mean AUC of insulin were dramatically improved in groups of DM1 and DM2(all P

Objective To explore the effect of using pedicle skin flaps for treatment of skin and soft tissue defects.Methods From January 2004 to September 2010,87 patients with skin and soft tissue injury which had been treated by using pedicle skin flaps without vessel anastomosis were retrospectively analyzed,including 61 males and 26 females with an average age of 40 years (range,14-61).The pedicle skin flaps selected were known vessel skin flaps or unknown vessel random skin flaps.The reconstruction treatment increased multiple selections of skin flaps for patients.Skin and soft tissue defects were located at the hand in 21 cases,at the lower extremities in 66.The kinds of skin flaps included adjacent transposition flaps in 50 cases,distal skin flaps in 37.The areas of the skin flaps were 2.0 cm×1.5 cm-18 cm×12 cm.Donor sites were as follows:lower leg in 61 cases,abdomen in 18,and hand in 8.Results Three patients appeared skin flaps hematoma after operation,which were removed immediately with no effect on flaps survival.Pedicle skin flap was compressed due to unstable fixation in 1 case,which made distal flap tissue necrosis approximately 2 cm in width.After dressing change for 6 weeks,the flaps got survival.Thirty seven patients underwent the second stage operation for breaking the perdicle 3 weeks after the first operation.All 87 patients were obtained follow-up for an average of 13 months (range,3-60).The total of 90.80％ patients treated with pedicle skin flaps were satisfied with the results.Conclusion Pedicle skin flap surgery can be performed more easily with less limitations of surgical designs.The survival rate of the flap is high,and the treatment of pedicle skin flaps could be well applied for soft tissue defects.