Type 1 diabetes mellitus (TlDM) is an autoimmune disease characterized by the destruction of pancreatic β cells, which sequentially leading to the reduction of insulin secretion. Numerous susceptibilities, including genetic and environmental factors, may be involved in triggering the T1DM. The ideal therapy for T1DM should focus on both preventing the immune system from attacking β cells, and also the stimulation of β cell neogenesis. In vitro and in vivo studies both demonstrated that GLP-1 analogues were able to promote the synthesis and secretion of insulin, stimulate neogenesis of β cells, increase β cells replication, and reduce β cell apoptosis. Hence, GLP-1 analogues have been shown to be potential treatment for T1DM. This work reviewed recent publications regarding to GLP-1 application in TlDM and pancreatic islet transplantations and prospected the future research directions.

Histone acetylation is a crucial part of histone modifications in epigenetics.Histone acetylation is involved in the onset of diabetes and diabetic complications,through the mechanism of inducing hyperglycemia by means of metabolic memory effect,interfering islet development and regulating the expression of inflammatory factors and pathogenic genes.Genome-wide association studies are gradually unveilling the pathogenesis of diabetes and preclinical studies are rapidly elucidating that histone deacetylase should be considered as a new target for the treatment of diabetes.

BACKGROUND: Jaw defects are common clinically. It is desirable to find ideal seed cells combined with scaffolds to construct tissue engineered jaws for curing these diseases. OBJECTIVE: To investigate the growth characteristics of bone marrow mesenchymal stem cells (BMSCs) transfected with basic fibroblast growth factor (bFGF) gene after seeded on coral scaffold in vitro. DESIGN, TIME AND SETTING: An experimental study of bone tissue engineering was performed in the Research Institute of Stomatology, Sun Yat-sen University between March 2006 and June 2008. MATERIALS: Natural coral from China Hainan bench was made into pieces of 8 mm×8 mm×2 mm. METHODS: BMSCs were isolated from New England rabbits by density gradient centrifugation and then purified by adherent separation. bFGF-pcDNA3 gene was transfected into BMSCs using Lipofectamine TM 2000. bFGF gene-transfected (transfected group) or untransfected (untransfected group)BMSCs were seeded on different coral scaffolds. In addition, bFGF gene-transfected BMSCs were simply cultured but not on the coral scaffold for control (simple culture group). MAIN OUTCOME MEASURES: BMSC proliferation was detected by methyl thiazolyl tetrazolium (MTT) assay and BMSC growth on coral scaffold was observed under the scanning electron microscope. RESULTS: MTT assay showed that the BMSC proliferation rate was significantly higher in the transfected group than in the untransfected group (P < 0.05) and that there was no significant difference in BMSC proliferation between the transfected and simple culture groups (P > 0.05). Scanning electron microscope results displayed that BMSCs adhered to and spread over the coral scaffold, exhibiting various appearances, with some cells had grown into scaffold micropores or spanned micropore surface, and some extracellular matrix secreted by BMSCs were found. CONCLUSION: The transfected group exhibited better growth of BMSCs transfected by bFGF gene than the untransfected group. These findings indicate that coral skeleton does not influence BMSC proliferation and can be used as a scaffold of BMSCs to construct tissue-engineered bone.

Objective To study the role of glucagon-like peptide-1 (GLP-1) analogue liraglutide played in the proliferation of CD4+CD25 T cells in normal people and newly-onset type 1 diabetic patients,and to evaluate the possible immune regulatory role of liraglutide in the therapy of type 1 diabetes.Methods CD4+ CD25-T cells of 10 normal people and 10 newly-onset type 1 diabetic patients were separated from peripheral blood by MACS immunomagnetic beads and stimulated by Human T-Activator CD3/CD28 Dynabeads to proliferate.CFSE labeling technique was used to evaluate the proliferation of CD4+ CD25-T cells by flow cytometry.Liraglutide of different concentrations(0,25,50,and 100 nmol/ml) was added to the proliferation system,then the proliferation of CD4+CD25-T cell was measured.Results (1) Liraglutide suppressed the proliferation of CD4+ CD25-T cells from either normal people or type 1 diahetic patients with dose-dependent manner (P ＜ 0.05).(2) Under the different concentrationsofliraglutide,the proliferation ofCD4+CD25 T cells from diabetic patients was mueh more robust than that of normal people (P＜0.01).(3) The inhibitory effects of liraglutide on CD4+ CD25-T cells proliferation in normal people and diabetic patients were similar (P＞0.05).Conclusion The proliferation of CD4+ CD25 T cells in type 1 diabetic patients was more robust than normal people,which indicated cellular immune dysfunction in type 1diabetes.Liraglutide inhibits the proliferation of CD4+ CD25-T cells of type 1 diabetic patients in vitro.The immunosuppression effect of liraglutide may have potential value in the treatment of type 1 diabetes.

Objective To decrease the p53 gene expression at cellular and animal levels in marmoset using RNA interference technique.Methods The shRNA interference sequences were designed and inserted into the adeno-associated virus vector plasmid after bioinformatics analysis.The plasmids were transfected into African green monkey kidney cos-7 cells.The suppression of p53 mRNA was detected by real-time PCR, and the changes of p53 protein expression were detected by Western bolt.The adeno-associated virus-8 was injected through the hind leg vein.The changes of p53 protein expression in the liver tissue was detected by Western blot and immunohistochemistry.Results We screened two RNA interference effective arget sequences.The expression of p53 mRNA was suppressed ( 82.7 ±8.1 )% and ( 80.7 ± 7.5)%, respectively (P<0.05), and the expression of p53 protein was decreased (77.3 ±11.5)% and (73.7 ± 10.7)%, respectively (P<0.05).The two marmosets after virus infection showed that there were virus distributions in the liver, testes, and neck detected by in vivo fluorescence imaging.The expression of p53 in the marmoset liver was detected by western blot, immunohistochemistry analysis showing no obvious changes.Conclusions In the present study, the decrease of P53 gene expression at cellular level is achieved, however, the liver P53 protein in the marmoset liver is not significantly changes.Further optimization of the way of infection is needed in the future.

Objective: To investigate the effects of buccal acupuncture on analgesia, immune indicators, and expression levels of Survivin and Livin proteins in patients with advanced-stage primary liver cancer. Methods: Eighty patients with advanced-stage primary liver cancer were selected and divided into control and treatment groups according to the difference in treatment modalities, with 40 patients in each group. The control group received transcatheter arterial chemoembolization (TACE), and the treatment group received buccal acupuncture in addition to TACE. The recent efficacy, analgesic effect, liver function, serum tumor markers, Survivin and Livin protein expression levels in liver cancer tissue, and immune indexes were analyzed and compared between the two groups. Results: The objective response rate (ORR) and disease control rate (DCR) of the treatment group were 37.5％ and 77.5％, respectively, which were significantly higher than those of the control group (22.5％ and 52.5％), and the recent efficacy of the treatment group was significantly better than that of the control group (P<0.05). The onset of analgesia in the treatment group was significantly faster than that in the control group (P<0.05), the duration of analgesia was significantly longer than that in the control group (P<0.05), and the numeric rating scale (NRS) score of pain after treatment was significantly lower than that in the control group (P<0.05). In the treatment group, the aspartate aminotransferase (AST), alanine aminotransferase (ALT), and albumin/globulin (A/G) were significantly lower than those in the control group (P<0.05), and the serum levels of alpha-fetoprotein (AFP), alpha-L-fucosidase (AFU), and carcinoembryonic antigen (CEA) were significantly lower than those in the control group (P<0.05), and the expression levels of Survivin and Livin in liver cancer tissue were significantly lower than those in the control group (P<0.05); CD4+ and CD4+/CD8+ in the treatment group were significantly higher than those in the control group, and CD8+ was significantly lower than that in the control group after treatment (P<0.05). Conclusion: Buccal acupuncture can reduce the degree of pain and liver function damage in patients with advanced- stage primary liver cancer and lower the serum tumor marker levels, and its mechanism of action may be related to the down-regulation of Survivin and Livin protein expression levels in the liver cancer tissue and the regulation of the immune function.

Objective To establish an indirect immunofluorescence assay for detection of murine norovirus ( MNV) .Methods Mouse leukaemic monocyte macrophage cell line RAW 264.7 cells were infected with MNV-1 and cultured for 36 hours to collect the virus and uninfected cells , and to make antigen glass slides .BALB/c mice were gavaged with MNV-1 (107 TCID50) and infected sera were collected as positive control .The serum was 1:10 diluted and used for measuring MNV antibody by immunofluorescence assay ( IFA ) .80 serum samples were tested using the two methods , IFA and ELISA, and the discrepant samples were validated by Western blotting .Results RAW264.7 cells were infected with MNV-1 for 36-48 h, showing an infection rate of 60% of the cells, and the cells infected for 36 h were preferred.IFA method was used to detect the serum with MNV-1 infection and showed that the antibody content was gradually increased at one week after infection , reaching a maximum antibody concentration at 4 weeks after infection , and maintained a stable level later .The mouse serum at four weeks after MNV-1infection was used as positive quality control . Among the 80 serum samples , 27 positive and 53 negative cases were detected by IFA method , and 32 positive and 48 negative cases were detected by ELISA .The five discrepant samples were verified by Western blotting , resulted in 3 positive and 2 negative cases . The coincidence rate of IFA was 96.0% and that of ELISA methods was 97.5%. Conclusions Basically, immunofluorescence assay can be used to detect the MNV-1 infection in mice, although false negative result may occur occasionally .IFA and ELISA detection can be selected as initial screening measures , and use Western blot assay to verify the discrepant samples .

Objective In order to establish a rhesus monkey model of p53 gene silencing, firstly we screened and determined the effective silencing targets of p53 gene at the cellular level in rhesus monkey.Methods The expression of p53 gene was detected in COS-7 cells ( derived from the kidney of the African Green Monkey, Cercopithecus aethiops).Three small hairpin RNA ( shRNA) sequences targeting rhesus monkey p53 gene were designed, analysed by bioinformatics, and inserted into lentivirus-based gene silencing constructs FUGW-TDT.The plasmids of p53-RNAi and control vector were transfected into the COS-7 cells, respectively.The suppression of p53 mRNA was detected by real-time PCR, and the changes of p53 protein expression were detected by Western blot assay.Results p53 gene expression was detected in COS-7 cells.Bioinformatics analysis showed that three gene-silencing sequences were screened which lied in the open reading frame ( ORF) region and targeted 238 -258bp, 681 -701bp, 169 -189bp of the rhesus monkey p53 mRNA.At 48 hrs after transfection of the three silencing constructs, p53 mRNA was suppressed by(87.17 ±4.03)%, ( 72.62 ±4.11)% and(76.22 ±0.98 )%, and p53 protein was suppressed by ( 84.44 ±2.18 )%, ( 71.04 ±1.18)% and ( 74.17 ±0.95 )%, respectively. Conclusions We obtained three effective target sequences showing high efficiency in p53silencing, which can be used in further studies on gene silencing in rhesus monkey.

Objective To explore the clinical applicating and efficacy of free fibula osteomyocutane- ous flap in mandible defect reconstruction in osteoradionecrosis patients.Methods The mandible defects were reconstructed by free fibula flaps with or without muscle cuff.The soft tissue defects were repaired by skin paddles.Status of osteotomy in fibula and flap survival was recorded.The complication in recipient site and donor site,as well as mouth opening and occlusion were reviewed.Facial contour and chewing function after reconstruction were evaluated.Results Patients were followed up 3-16 months.4 free fibula flaps with muscle cuff and 5 without muscle cuff survived well.The size of mandible defects covered from 6cm to 17cm. And the harvested fibula flaps with length of 8.6-17cm were cut into 3 segments in 2 cases,and 2 segments in 5 cases.Fibula flap was divided into 2 segments and overlapped in 2 cases.No serious complication was oh- served in recipient site and donor site.Satisfying esthetic result and normal occlusiong of heath mandible were obtained in all cases.The degree of mouth opening was 2.5-3.3cm.Fair chewing function was revealed in re- constructive region after prosthesia repaired.Conclusion Free fibula osteomyocutaneous flap is relatively ideal reconstruction material of mandible defect in osteoradionecrosis patients for its high survival rate and well esthetic results.

Objectives To explore the effect of transient continuous subcutaneous insulin infusion (CSII) on β cell function, insulin resistance and vascular endothelial injury in newly diagnosed type 2 diabetic patients and its potential mechanism. Methods Ten patients with newly diagnosed type 2 diabetes mellitus (T2DM) accepted CSII for two weeks. Intravenous glucose tolerance test (IVGTT) and hyperinsulinemia euglycemia clamp test were performed before and after CSII. Serum soluble E-selectin (sE-selectin) was used to evaluate the injury of vascular endothelial cell, while serum high sensitivity C reactive protein (hsCRP) and soluble CD_(14) (sCD_(14)) were both used to assess inflammatory condition. Results (1) Compared with those before treatment, the blood glucose levels of IVGTT, the area under the curve of the blood glucose, glycosylated hemoglobin, TC and LDL-C in the patients were decreased after CSII (P < 0. 05 or 0. 01). (2) Compared with those before treatment, the insulin levels of IVGTT (except the fasting insulin), the area under the curve of insulin and acute insulin response were all increased after CSII(P < 0.05 or 0.01). (3) Compared with that before treatment, the glucose infusion ratio in the clamp test [(3.46±1.66)mg·kg~(-1)·min~(-1) increased to (7.14±2.37)mg·kg~(-1)·min~(-1)]and HOMA-β elevated, while HOMA-IR declined (P <0. 05 or 0. 01 in all). (4) Compared with those before treatment, the levels of serum sE-selectin, sCD_(14) and hsCRP were decreased (P < 0. 01, except for hsCRP) . Conclusion Transient intensive insulin therapy in patients with newly diagnosed T2DM is useful to restore 13 cell function, attenuate insulin resistance, repair vascular endothelial injury and improve the disorder of blood sugar and lipid. The mechanism may be related with the inhibition of inflammation in patients.