OBJECTIVE:To observe the clinical efficacy and safety of saxagliptin combined with metformin in the treatment of type 2 diabetes. METHODS:A total of 95 patients with type 2 diabetes were randomly divided into observation group(47 cases) and control group(48 cases). All of the patients received Metformin hydrochloride sustained-release tablets 0.5 g for continuous 4 weeks,orally,3 times a day;based on the treatment,control group was continuously given Metformin hydrochloride sustained-re-lease tablets 0.5 g for continuous 24 weeks,orally,3 times a day;observation group was given Saxagliptin tablets 5 mg continu-ous 24 weeks based on the treatment of control group,orally,once a day;the treatment course was 28 weeks. The clinic data was observed,including clinical efficacy,FPG,2 h PG,bed time glucose,HbA1c,BMI before and after treatment,incidence of hypoglyce-mia,severe hypoglycemia and adverse reactions. RESULTS:The total effective rate in observation group was higher than control group,the incidence of hypoglycemia in observation group was lower than control group(P<0.05). After treatment,the glucose in-dexes in 2 groups was significantly lower than before,and FPG and HbA1c in observation group were lower than control group(P<0.05). There were no significant differences in the BMI before and after treatment,incidence of severe hypoglycemia and adverse reactions between 2 groups(P>0.05). CONCLUSIONS:Saxagliptin combined with metformin has better efficacy and safety than metformin alone in the treatment of type 2 diabetes,and can significantly reduce the incidence of hypoglycemia.

Adolescent ; Child ; Child, Preschool ; Core Binding Factor Alpha 3 Subunit ; genetics ; metabolism ; DNA Methylation ; Female ; Humans ; Infant ; Lymphoma ; genetics ; metabolism ; Male ; Promoter Regions, Genetic

Objective To investigate the influence of CD4+CD25+ regulatory T cells (Tregs) on the disease progression in MRL/lps mice. Methods Tregs were separated by using magnetic beads from splenic cells of MRL/lps mice and BALB/c mice, and concentrated. Twenty-four MRLAps mice were equally divided into 3 groups, test group 1 injected with Tregs from MRL/lps mice, test group 2 injected with Tregs from BALB/c mice, and control group injected with physiological sodium chloride solution. Three weeks later, the levels of urine protein as well as serum anti-dsDNA antibody were determined; subsequently, the mice were sacrificed followed by histopathological and immunopathological examination of renal tissue. Results A significant decline was observed in the test group 1 compared with the test group 2 and control group in the urine protein score (10.63 ± 4.17 vs. 20.00 ± 5.35 and 18.75 ± 8.34, both P < 0.05), serum anti-dsDNA antibody level (5.36 ± 2.40 pg/ml vs. 9.57 ± 1.97 pg/ml and 10.75 ± 3.98 pg/ml, both P < 0.05), glomerular sclerosis index [(32.00 ± 12.09)% vs. (45.50 ± 13.68)% and (47.50 ± 10.78)%, both P< 0.05], and immunofluorescence intensity of IgG immune complex in renal tissue (1.88 ± 0.99 vs. 2.88 ± 0.64 and 2.75 ± 0.71, both P< 0.05). No significant difference was noted in renal tubule interstitial impairment index between the 3 groups (4.63 ± 1.92, 6.00 ± 1.07 and 5.75 ± 1.28, all P> 0.05). There was no statistical difference between the test group 2 and control group in terms of any of the above parameters (all P > 0.05). Conclusions Injection of Tregs from homologous mice could significantly down-regulate proteinuria degree, serum anti-dsDNA antibody level, glomerular sclerosis index and IgG immune complex level in renal tissue, and thereby decelerate the progression of renal impairment in MRL/lps mice.

ObjectiveTo investigate the mental health of freshmen of university.MethodsSymptom Checklist 90 (SCL-90) was used to self evaluate 7763 freshmen of university in Ningbo.Results and ConclusionThe overall level of the mental health of this group was higher than that of the Chinese module, and the prevalence of psychological symptoms was 13.2%. The average level of mental health of female were inferior to male. The difference between the students from cities or towns and from the countryside were distinct. Students who were satisfied with their majority get noticeably higher average score than the ones who were not satisfied.

OBJECTIVETo investigate the methylation status of zonula occludens-1 (ZO-1) gene promoter and its clinical significance in children with stage IV non-Hodgkin lymphoma (NHL) and to provide a basis for further etiological study and early diagnosis of this disease.

METHODSFifty-five children with a confirmed diagnosis of stage IV NHL (40 cases of T-NHL and 15 cases of B-NHL) were selected as the case group, and 20 children with diseases other than hematologic malignancies were selected as the control group. Bone marrow samples were collected from these subjects. Methylation-specific PCR (MS-PCR) was applied to evaluate the methylation status of ZO-1 gene promoter, and the integrated optical density (IOD) was determined. RT-PCR was used to measure the mRNA expression of ZO-1.

RESULTSMS-PCR showed that the methylated bands of ZO-1 gene promoter were found in 39 (70.9%) of 55 patients in the case group before treatment, while no ZO-1 gene promoter methylation was detected in the control group. With close tracking of 47 cases in the study group, consisting of 32 cases of T-NHL and 15 cases of B-NHL, the rates of ZO-1 gene promoter methylation prior to treatment were 72% and 67%, respectively, (P>0.572). The cases of T-NHL and B-NHL showed no significant changes in methylation rate in the early and middle phases of chemotherapy (P>0.05), but they showed significant changes in methylation rate in the late phase of chemotherapy (P<0.05). RT-PCR showed that the NHL cases carrying methylated ZO-1 gene had no mRNA expression of ZO-1, while all children in the control group had mRNA expression of ZO-1. There was no linear relationship between the total number of peripheral blood leukocytes and ZO-1 gene IOD (r=0.093, P=0.575); a positive correlation was found between the number of malignant cells in bone marrow and ZO-1 gene IOD (r=0.669, P<0.001).

CONCLUSIONSZO-1 gene shows a hypermethylation status in children with NHL, and the methylation level is positively correlated with the number of malignant cells in bone marrow. ZO-1 may be used as a novel molecular marker in early diagnosis, outcome assessment, prognostic evaluation, and detection of minimal residual disease.

Adolescent ; Child ; Child, Preschool ; DNA Methylation ; Female ; Humans ; Infant ; Lymphoma, Non-Hodgkin ; genetics ; Male ; Promoter Regions, Genetic ; Zonula Occludens-1 Protein ; genetics

BACKGROUND:The high level of matrix metalloproteinase 9 (MMP9) is thought to slow down the healing of diabetic foot ulcers. Whether it can influence the biological behaviors of skin fibroblasts and affect wound healing is stillunclear. The present study aimed to observe changes in the biological behaviors of rat dermal fibroblasts induced by high expression of MMP9 and to clarify the possible mechanisms of wound healing for diabetic foot. METHODS:A cellmodel of skin fibroblast with high expression of MMP9 was established by co-culture of high glucose (22.0 mmol/L) and homocysteine (100 μmol/L). A control group was incubated with normal glucose (5.5 mmol/L). Realtime PCR, ELISA and gelatin zymography were used to detect the MMP9 mRNA, protein expression and activity of MMP9. Flow cytometry, CCK-8, ELISA assay, scratch test and transwellwere used to detect cellproliferation, viability, collagen (hydroxyproline) secretion, horizontal migration and vertical migration of cells. The data were expressed as mean±SD. P value less than 0.05 was considered statistically significant. RESULTS:The expression of MMP9 mRNA, protein levels and the activity of MMP9 were much higher in the high MMP9 group than in the control group (7.05±1.02 vs. 1.00±0.00, 206.9±33.6 pg/mL vs. 40.4±5.9 pg/mL, and 1.47±0.13 vs. 0.57±0.12, respectively,P<0.01). The proportion of S-phase cells, proliferation index, cellviability, collagen (hydroxyproline) secretion, horizontal migration rate and the number of vertical migration cells were lower in the high MMP9 group than in the control group (P<0.01). CONCLUSION:Fibroblasts with a high expression of MMP9 decreased proliferation, activity, secretion and migration of collagens, suggesting that MMP9 may inhibit the biological behaviors of fibroblasts.

OBJECTIVETo explore the correlation between the HLA-DR13, basic core promoter (BCP), changes of T lymphocyte subset and clinical Chinese medical syndromes of chronic hepatitis B (CHB).

METHODSTotally 102 CHB patients were syndrome typed as Gan depression Pi deficiency syndrome (GDPDS), Pi-Shen yang deficiency syndrome (PSYDS), Gan-gallbladder dampness heat syndrome (GGDHS), Gan-Shen yin deficiency syndrome (GSYDS), and static blood blocking collaterals syndrome (SBBCS). Besides, 30 healthy subjects were recruited as the normal control group. The blood HBV-DNA level and HLA-DR13 gene were detected with real time fluorescent PCR. The expression of CD4+ and CD8+ in T lymphocytes was detected using flow cytometry. The mutation of serum A1762T/G1764A was detected using PCR sequencing. Hepatitis Be antigen (HBeAg) was detected with ELISA, and correlation between various Chinese medical syndrome types and objective indicators were analyzed.

RESULTSThere was no statistical difference in HBV-DNA quantitative results among various syndrome types (P > 0.05). HBeAg positive rate was higher in GDPDS than in other syndrome types (P < 0.05). It was sequenced as GDPDS > GSYDS > SBBCS > GGDHS > PSYDS. Compared with the normal control group, percentages of CD3+ and CD3+ CD4+ were lower in PSYDS (P < 0.05). The ratio of CD3+ CD4+/CD3+ CD8 was lower in GGDHS and PSYDS than in the normal control group (P < 0.05). There was no statistical difference in the CD3+ CD8+ percentage among various syndrome types (P > 0.05). The quantitation of HLA-DR13 gene was lower in GDPDS and GSYDS than in the normal control group (P < 0.05). The positive rate of BCP mutation was higher in GSYDS than in other syndrome types (P < 0.05).

CONCLUSIONCo-detection results of HLA-DR13 and BCP could be used as reference indices of Chinese medical syndrome typing of CHB.

HLA-DR Serological Subtypes ; genetics ; metabolism ; Hepatitis B, Chronic ; classification ; diagnosis ; genetics ; Humans ; Medicine, Chinese Traditional ; Promoter Regions, Genetic ; Syndrome ; T-Lymphocyte Subsets ; metabolism ; Yang Deficiency ; Yin Deficiency

OBJECTIVETo investigate the effect of Oviductus Ranae (OR) on the expressions of CyclinD1, CDK6 and P15 in the liver of aged male rats.

METHODSEighteen male SD rats were randomly divided into 3 equal groups, namely the OR group, VE group and ageing model group. The rats received subcutaneous injection of D-galactose for 6 weeks to establish the aging models, and another 6 rats were injected daily with normal saline (NS) to serve as the normal control group. From the third week of the experiment, the rats were given oral OR or Vitamin E (VE) accordingly till the sixth week. After completion of the drug administration, all the rats were sacrificed for detecting the expressions of CyclinD1, CDK6 and P15 in the liver tissue by Western blotting.

RESULTSThe relative expression levels of CyclinD1, CDK6 and P15 in the liver of the rats in the OR group were 41.73-/+0.54, 23.29-/+0.30 and 1.49-/+0.30, respectively, significantly up-regulated as compared with those in the ageing model group (P<0.01). The expressions of the proteins were obviously down-regulated in the model group in comparison with those in the normal control group.

CONCLUSIONSOR treatment can lower the expressions of Cyclin D1 and CDK6 in the liver to enhance the liver cell proliferation in aged male rats. OR also promotes the expression of P15 through a feedback mechanism to prevent excessive proliferation of the cells. The effect of OR against ageing is mediated possibly by up-regulation of the proteins associated with the cell proliferation in the liver, a mechanism different from that of VE.

Aging ; metabolism ; Animals ; Cyclin D1 ; metabolism ; Cyclin-Dependent Kinase 6 ; metabolism ; Cyclin-Dependent Kinase Inhibitor p15 ; metabolism ; Liver ; metabolism ; Male ; Materia Medica ; pharmacology ; Rats

Objective To study and discuss what part does methylated Id4 gene participate in malignant lymphoma stage Ⅳ by detecting the extent of how much Id4 gene has been methylated in afflicted children who suffer from malignant lymphoma.Methods Forty-two patients who had diagnosed with malignant lymphoma [Hodgkin's disease (HD),Non-Hodgkin's lymphoma(NHL)] were selected as study group.Their chemotherapy stages of pre-treatment,early-treatment,mid-treatment,post-treatment and clinical remissions or relapse throughout the entire treatment had been traced.At each stage the expression of methylated Id4 gene mRNA was detected by methylation-specific polymerase chain reaction (MS-PCR) and compared with the control group.The control group consisted of 20 non-neoplastic hematologic disorder affected children as sample donors.Results MS-PCR detection:in pre-treatment stage,there were 27 patients who were found methylated or partially methylated Id4 genes.Methylated ratio was thus at 64.3％ (27 patients out of a total of 42 patients).Those 27 patients were actively traced down along with different stages of treatment (21 NHL patients,6 HD patients).Before NHL there was 55.6％ methylated Id4 gene (15 cases out of 27 NHL patients).During chemotherapy treatment,there was 51.9％ of methylated Id4 gene positive (14 cases out of 27 patients).In post chemotherapy treatment,there was 48.1％ of methylated Id4 gene positive (13 cases out of 27 patients).Totally there were 9 patients showed clinical recovery after chemotherapy.There was 44.4％ of traceable methylated Id4 gene after recovered chemotherapy patients (4 recovered patients still carrying positive reading of methylated Id4 gene out of totally 9 recovered patients).There were 2 patients relapsed,with traceable methylated Id4 gene re-appeared in them afterwards.Throughout different treatment stages,there was no significant correlation in the treatment result and the appearance of methylated Id4 gene in early treatment stages (all P ＞ 0.05).But in latter treatment of chemotherapy,the correlation started to emerge (P ＜ 0.05) ; The overall statistics on NHL and HD share the same statistical pattern on different stages (all P ＞ 0.05).The control group had 20 patients,none of them had methylated Id4 gene.The study group showed noticeable difference in methylated Id4 gene before and after the experiment (P ＜ 0.001).RT-PCR result showed that before chemotherapy treatment,all of those who carried methylated Id4 gene had no expression of mRNA,by comparison to the control1 group which all had expression of mRNA.Conclusions Methylated Id4 gene is closely related in affected children who suffer from malignant lymphoma and its complications.The expression of Id4 gene is depressed when it has been methylated.The state of methylated Id4 gene is changed as patient's condition changed,so the methylated Id4 gene is thus a possible indicator of early diagnostic tool for children lymphoma.

OBJECTIVETo investigate the methods and solve the technical bottlenecks in the preparation of recombinant human protein hZP3 using the baculovirus expression system and pave the technical ground for the production and application of recombinant hZP3.

METHODSThe recombinant vector pFASTBAC HTa-hZP3 was constructed and transferred to competent E. coli cells carrying bacmid to produce recombinant bacmid by homologous recombination. Sf9 cells were transfected with the recombinant bacmid to produce recombinant baculovirus. Full-length recombinant hZP3 (amino acids 1-424) and truncated recombinant hZP3 (amino acids 23-348) were expressed in the sf9 cells by infection with the recombinant baculovirus. The expression time of hZP3 was determined by Western blot and its purification was explored.

RESULTSThe recombinant bacmid and baculovirus were successfully constructed for expressing both the full-length and truncated hZP3. The maximal expression of recombinant hZP3 in the sf9 cells was achieved at 72-96 hours after baculovirus infection. Some of the recombinant hZP3 with His-tag could bind affinity matrix and got purified but most of the solubilized hZP3 passed through and the reasons remained unknown. Purified recombinant hZP3 labeled with Dylight Dye488 was able to bind human sperm.

CONCLUSIONIt is feasible to express recombinant hZP3 in insect cells using the baculovirus system though the yield of hZP3 needs to be optimized. The methods for efficient enrichment and purification of recombinant hZP3 require further exploration.

Baculoviridae ; genetics ; metabolism ; Blotting, Western ; Egg Proteins ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Humans ; Membrane Glycoproteins ; genetics ; metabolism ; Receptors, Cell Surface ; genetics ; metabolism ; Recombinant Proteins ; genetics ; metabolism ; Transfection ; methods ; Zona Pellucida Glycoproteins