0.05). The inducing action of glucose to the apoptsis of MC3T3-E1 osteoblast was in concentration and time dependent manner(P

Objective To investigate the effects of miR-335-5p on the proliferation and apoptosis of osteoblasts which were exposed to high glucose condition, and explore its possible molecular mechanisms. Methods MC3T3-E1 osteoblasts were divided into four groups:control group(5. 5 mmol/L glucose), high glucose group(HG group, 22. 0 mmol/L glucose), agomir-335-5p group(transfected with agomir-335-5p and exposed to 22. 0 mmol/L glucose) , and agomir negative control group( agomir NC group, transfected with agomir negative control and exposed to 22. 0 mmol/L glucose), cultured for 7 days. Cell proliferaton, cell apoptosis, expressions of miR-335-5p and dickkopfhomolog1(DKK1)mRNA,proteinlevelsofDKK1andcysteinylaspartate-specificproteinase-3(caspase-3) were detected using MTT, flow cytometry, quantitative realtime PCR and western blot, respectively. Results Compared with control group, the expression of miR-335-5p mRNA and cell proliferation in HG group were significantly decreased(P<0. 05), while cell apoptosis and the protein levels of DKK1 and caspase-3 were increased significantly(P<0. 05), the expression of DKK1 mRNA did not change (P>0. 05). The miR-335-5p mRNA expression and cell proliferation in agomir-335-5p group were higher than those in HG group and agomir NC group(P<0. 05). However, Cell apoptosis and the protein levels of DKK1 and caspase-3 in agomir-335-5p group were lower than those in HG group(P<0. 05). Conclusion High glucose inhibits the proliferation and induce the apoptosis of MC3T3-E1 osteoblast through decreasing the expression of miR-335-5p and subsequently increasing the DKK1 expression.

Objective To investigate the changes of bone mineral density (BMD) and its causing factors in menopausal female patients with type Ⅱ diabetes mellitus (DM2). Methods Dual energy X ray absorptimetry was used to measure the BMD of lumbar spines (L 2-4 ) and proximal femur in 82 female patients with type Ⅱ diabetes and 46 normal controls. According to BMD, DM2 patients were divided into two groups (DM A and DM B). Fasting plasma glucose (FPG), fasting plasma insulin (FINS), fasting plasma C peptide (FCP), glycated hemoglobin (HbAc1), urine albumin excretion rate (UAER), and urine ? 2 microglobin (? 2 MG) were measured and compared between the two groups. Results BMD of lumbar spines (L 2-4 ), femoral neck and Ward's triangle in diabetic patients were significantly lower than that of the controls ( P

Objective To examine the role of p38MAPK in high glucose-induced apoptosis of osteoblast MC3T3-E1 cell line, and to investigate its effect on the expressions of apoptosis-related molecules including caspase3, bax, and bcl-2. Methods The lentiviral vector containing short hairpin RNA targeting p38MAPK was constructed. The cultured osteoblast MC3T3-E1 cell were divided into 5 groups:normal control group(A group), high glucose group(B group), p38MAPK-shRNA transfection group(C group), signal transduction inhibitor group(D group), and transfection with negative control siRNA group(E group). RT-PCR was used to determine the p38MAPK mRNA expression levels in MC3T3-E1 cells. Flow cytometry(FCM)was employed to detect the cell apoptotic percentage. The protein levels of apoptosis-related molecules p38MAPK, p-p38MAPK, caspase-3, bax, and bcl-2 were assayed by Western blot. Ultrastructural alternation of MC3T3-E1 cell was observed under transmission electron microscopy(TEM). Results The lentiviral vector containing short hairp in RNA targeting p38MAPK was successfully constructed and transfected into MC3T3-E1 cells. RT-PCR result suggested that the siRNA targeting p38MAPK could effectively reduce the p38MAPK mRNA expression level induced by high glucose in MC3T3-E1 cell line. FCM showed siRNA significantly decreased high glucose-induced apoptosis percentage of MC3T3-E1 cells(P＜0.01). Meanwhile, we also found the siRNA significantly attenuated the proteins levels of p38MAPK, p-p38MAPK, caspase-3, and gene bax induced by high glucose in MC3T3-E1 cells, whereas the protein level of gene bcl-2 was enhanced remarkably when compared with high glucose group and negative control siRNA group(P＜0.01, P＜0.05).Conclusion The iRNA targeting p38MAPK suppressed high glucose-induced MC3T3-E1 cell apoptosis via inhibiting the activation of p38MAPK signaling pathway, thereby reducing the expressions levels of p-p38MAPK, caspase-3 and gene bax, and up-regulating the level of gene bcl-2.

SD rats were injected with streptozotocin and fed with high fat diet,used as type 2 diabetic model. Transcription factor Fox01 [in nucleus (15.00±1. 15 vs 6.45±0. 62) %, P<0.05], Caspase-3 [(23.73 ±1.48 vs 6.30±2.20)% ,P<0.01] expressions in pancreatic istet β cells and the positive rate of islet β cells apoptosis[(22.29±1.84 vs 6.25±2.42) %, P<0.01] in diabetic rats were higher than those of control rats. The islet cells which highly expresed Fox01 (in nucleus)and caspase-3 were just the apoptotic islet cells. Therefore,Fox01 may be involved in regulating apoptosis of islet β cells in type 2 diabetes.

Objective To investigate the relationship between angiotensin Ⅱ and pancreatic islet β cell secretion function under different glucose tolerance statuses. Method Forty-two patients with newly diagnosed type 2diabetes mellitus ( DM group), 38 subjects with impaired fasting glucose/impaired glucose tolerance ( IFG/IGTgroup) ,and 40 normal control subjects (NGT group) underwent intravenous glucose tolerance test. Fasting plasma angiotensin Ⅱ ( Ang Ⅱ ) and adiponectin were assayed by ELISA. Acute insulin response from 3 to 10 min( AIR3-10 ),the area under the curve( AUCⅠ ) and the peak concentration of the first-phase ( 0-10 min) insulin secretion, the area under the curve of the second-phase( 10-120 min) insulin secretion( AUCⅡ), homeostasis model assessment for β cell function index(HOMA-β) and homeostasis model assessment for insulin resistance index(HOMA-IR) were calculated to explore the relationship with Ang Ⅱ. Result ( 1 ) The levels of Ang Ⅱ in DM group and IFG/IGT group were significantly higher than that in NGT group( P＜0.05 ). The AIR3-10, AUCⅠ and peak concentration, AUCⅡ ,adiponectin in DM group and IFG/IGT group were significantly lower than those in the NGT group ( P＜0. 05), and these results were more significantly reduced in DM group compared with those in IFG/IGT group. (2) Ang Ⅱ was negatively correlated with AIR3-10, AUCⅠ and the peak concentration, AUCⅡ, adiponectin, HOMA-β ( P＜0. 01 ), and positively correlated with fasting blood glucose,2 h blood glucose after glucose loading, fasting insulin, HOMA-IR (P＜0. 05 ). (3)Multiple stepwise regression analysis showed that Ang Ⅱ was independently associated with AUCⅠ and AUCⅡ.Conclusion Ang Ⅱ was an independent factor that affected the insulin secretion function of pancreatic islet βcells. Ruling out the effect of blood pressure, body position, drugs, and other factors, high levels of Ang Ⅱ could predict the dysfunction of pancreatic islet β cell as well as insulin resistance in patients with type 2 diabetes.

The relationship between oxidative stress and the first-phase of pancreatic β cell insulin secretion in subjects with different statuses of glucose tolerance was explored. Fasting adiponectin, 8-hydroxydeoxyguanosin ( 8-OHdG), malondialdehyde ( MDA ), superoxide dismutase ( SOD ), insulin area under the curve ( AUC ) from 0 to 10 min, and AIR3-5 were measured. The levels of oxidative stress-related markers were elevated, the activities of superoxide dismutase,adiponectin, and first-phase of insulin secretion were reduced with the disease progression. MDA and 8-OHdG were negatively correlated with adiponectin, homeostasis model assessment β cell function index ( HOMA-β ),AUC ,and AIR3-5. SOD was positively correlated with adiponectin, HOMA-β, AUC, AIR3-5. The plasma 8-OHdG and SOD were independently associated with AIR3-5. Oxidative stress exerts a significant effect on the first phase of pancreatic β cell insulin secretion, which may contribute to the pathogenesis of type 2 diabetes.

Objective To explore the relationship between the women menopausal period and metabolic syndrome (MS) .Meth-ods The female residents of Chongqing urban areas above 40 years were selected as the investigation group ,all the subjects were performed the questionnaire survey and the physical examination ,at the same time the biochemical indexes were detected .Finally , 1402 women of natural menopause were included in this study .The study subjects were divided into different groups according to the menopausal period of <5 years ,5- <10 years ,10- <15 years and ≥15 years .The Logistic regression analysis was adopted to analyze the relationship between the menopausal period with MS and its components .Results The MS prevalence in this group was 40 .87% ,and the menopausal period of the women with MS was significantly higher than that without MS (P<0 .05) .The MS prevalences of postmenopausal women in the menopause period of <5 years ,5 - <10 years ,10 - <15 years and ≥15 years were 29 .37% ,34 .29% ,45 .30% and 49 .13% respectively (P<0 .05) .After adjustment for age and BMI (except central obesity ) ,the MS risk in the menopausal period of 10- <15 years and was 1 .54 times of that in the menopausal period of <5 years .The Logis-tic regression analysis showed that BMI and menopausal period were the influence factors of MS .Conclusion Post-menopausal women are the high-risk group of MS and the menopausal period is correlated to MS .

Objective To test the RET-proto-oncogene in a multiple endocrine neoplasia type 2( MEN2) family for confirming the diagnosis and classification , guiding treatment and prevention , and improving the prog-nosis.Methods There were 2 patients of MEN2 with clinical diagnosis and 1 asymptomatic first-degree relative in the pedigree .PCR and direct gene sequencing of PCR produces were used to scan the entire 21 exons of RET-proto-oncogene in the 3 members of the pedigree and 3 normal controls .Results The 2 patients and 1 asympto-matic first-degree relative in the pedigree all had a mutation of the codon 634 in exon 11.It was a heterozygous missense mutation C634R(TGC → CGC).The 3 normal controls showed no abnormalities .Conclusion The gene test of RET proto-oncogene helps to confirm the diagnosis of the pedigree as MEN 2, which can guide the treatment and help identify one asymptomatic mutation carrier .

Objective To evaluate the efficacy of transarterial chemoembolization (TACE) and percutaneous cryosurgery sequential therapy for unresectable hepatocellular carcinoma (HCC).Methods Four hundred and twenty patients with unresectable HCC were divided into sequential TACE-cryosurgery sequential (sequential) group (n=290) and cryosurgery alone (cryoalone) group (n = 130). TACE was performed with the routine operation; the percutaneous cryosurgery was conducted 2 to 4 weeks after TACE. The patients were followed up at the first month and once every 2 to 3 month later. Liver ultrasound or both computer tomography and alpha fetal protein were examined during follow-up. Results During a mean follow-up of (42±17) months (range from 24 to 70 months), the local recurrence rate of ablated lesion was 17% for all the patients, 11% and 24% for patients in sequential group and cryoalone groups respectively (P=0. 001). The overall 1-, 2-, 3-, 4-and 5-year survival rate was 72%, 57%, 47%, 39% and 31%, respectively. The 1- and 2-year survival rates (71% and 61 % ) in sequential group were similar to those (73 % and 54 % ) in cryo-alone group (P=0.69 and 0. 147), while the 4- and 5-year survival rates were higher in sequential group (49 % and 39 % ) than those (29 % and 23 % ) in cryo-alone group (P= 0.001). Eighteen patients with large HCC (＞5 cm in diameter) in sequential group survived for more than 5 years while no one in cryo-alone group. Complication rate was 24% in all patients, 21% and 26% for the sequential and cryo-alone groups respectively (P=0. 06). The incidence of hepatic bleeding was higher in cryo-alone group than in sequential group (P=0. 02). Liver crack occurred in two patients of the cryoalone group. Conclusions Pre-cryosurgical TACE increased the cryoablation efficacy and decrease its complications, especially hepatic bleeding. TACE and cryosurgery sequential therapy may be a better treatment for unresectable HCC, especially for large HCC.