Circ_0000105 promotes liver cancer by regulating miR-498/PIK3R1.

BACKGROUND: A growing number of studies demonstrate that circular RNA (circRNA) has a prominent role in the regulation of numerous biological behaviors of tumor cells. The present study aimed to investigate the expression, function and molecular mechanisms of circ_0000105 in liver cancer. METHODS: A quantitative real-time polymerase chain reaction (qRT-PCR) was utilized to obtain the expressions of circ_0000105 and miR-498 in liver cancer tissues and cells. The association between the expression level of circ_0000105 and the clinicopathological characteristics of liver cancer patients was analyzed. qRT-PCR and western blotting were utilized to investigate the expression of phosphoinositide-3-kinase regulatory subunit 1 (PIK3R1) in cells. Cell counting kit-8, bromodeoxyuridine and flow cytometry assays were utilized to determine liver cancer cell proliferation and apoptosis, respectively. Bioinformatics, a luciferase reporter gene experiment and a RNA immunoprecipitation experiment were conducted to predict and confirm the targeting correlation of circ_0000105 and miR-498, as well as miR-498 and PIK3R1. RESULTS: Circ_0000105 was highly expressed in liver cancer tissues and cell lines. Moreover, its high expression was associated with an increase in the T stage of liver cancer patients and a low degree of tumor differentiation. Circ_0000105 overexpression promoted liver cancer cell proliferation and inhibited apoptosis, whereas knocking down circ_0000105 triggered the opposite effect. miR-498 was a downstream target of circ_0000105, inhibiting liver cancer cell proliferation and promoting apoptosis. Mechanistically, circ_0000105 indirectly up-regulated the expression of PIK3R1 by adsorbing miR-498. CONCLUSIONS: Circ_0000105 plays a cancer-promoting role in liver cancer by regulating the miR-498/PIK3R1 axis.

SOX18 Affects Cell Viability, Migration, Invasiveness, and Apoptosis in Hepatocellular Carcinoma (HCC) Cells by Participating in Epithelial-to-Mesenchymal Transition (EMT) Progression and Adenosine Monophosphate Activated Protein Kinase (AMPK)/Mammalian Target of Rapamycin (mTOR).

BACKGROUND Hepatocellular carcinoma (HCC) is one of the most common malignancies around the world. It has been verified that the expression of SOX18 is correlated to poor clinical prognosis in patients with ovarian cancer, non-small cell lung cancer, or breast invasive ductal carcinoma. However, the expression pattern and biological function of SOX18 in HCC tissues remains unclear. In this study, we set out to investigate the associated biological function and potential molecular mechanism of the SOX18 gene in HCC cells. MATERIAL AND METHODS The mRNA and protein expression levels of experimental related genes were detected by real-time polymerase chain reaction and western blotting assay, respectively. The MTT method was used to assess cell viability, and cell apoptosis analysis was performed by means of FACScan flow cytometry. Wound-healing assay and Transwell analysis were performed to evaluate the ability of cell migration and invasiveness, respectively. RESULTS SOX18 was highly expressed in various HCC cell lines. In addition, SOX18 promoted cell viability, migration, and invasion and simultaneously induce cell apoptosis. SOX18 promoted epithelial-to-mesenchymal transition (EMT) progression, and SOX18 downregulation activated the autophagy signaling pathway AMPK/mTOR in HCC cells. CONCLUSIONS SOX18 downregulation in HCC cells suppressed cell viability and metastasis, induced cell apoptosis and hindered the occurrence and progression of tumor cells by participating in the EMT process and regulating the autophagy signaling pathway AMPK/mTOR.

Relationship Between Interleukin-10 Gene C-819T Polymorphism and Gastric Cancer Risk: Insights From a Meta-Analysis.

BACKGROUND As a pleiotropic cytokine, interleukin-10 (IL-10) plays a regulatory role in carcinogenesis and tumor growth. The aim of this meta-analysis was to assess the susceptibility of the IL-10 gene C-819T polymorphism to gastric cancer. MATERIAL AND METHODS Study identification and data extraction were independently completed by 2 authors. Odds ratios (OR) and 95% confidence intervals (95% CI) were calculated and summarized. RESULTS In total, 11 articles including 1960 gastric cancer patients and 3705 controls were qualified. Overall analyses revealed a 13% reduced risk of gastric cancer conferred by the -819T allele relative to the -819C allele (OR=0.87; 95% CI: 0.77-0.97; P=0.016), without heterogeneity (I2=35.1%). In subgroup analyses, a significant difference was identified in East Asian populations (OR=0.85; 95% CI: 0.73-0.98; P=0.029, I2=43.6%), for gastric adenocarcinoma (OR=0.80; 95% CI: 0.66-0.96; P=0.017, I2=0.0%), and in population-based studies (OR=0.81; 95% CI: 0.70-0.93; P=0.003, I2=0.0%). The visual funnel plots and Egger's tests suggested no evidence of publication bias. CONCLUSIONS Extending previous findings, we demonstrate a protective role of the IL-10 gene -819T allele in susceptibility to gastric cancer, and this role was more evident for gastric adenocarcinoma.

Tumor Necrosis Factor-Alpha and 8-Hydroxy-2'-Deoxyguanosine are Associated with Elevated Urinary Angiopoietin-2 Level in Type 2 Diabetic Patients with Albuminuria.

BACKGROUND/AIMS: We previously showed that urine and serum Angiopoietin-2 (Ang-2) levels increased significantly with the degree of albuminuria in diabetes patients, but the reasons remain unclear. Consequently we aimed to determine whether there was an association between Ang-2, inflammatory cytokines (TNF-α and IL-18) and reactive oxygen species (8-OHdG and SOD) in type 2 diabetes patients with albuminuria. METHODS: This retrospective study evaluated 113 patients with type 2 diabetes and normoalbuminuria, microalbuminuria, or macroalbuminuria and 30 healthy controls. Serum and urine TNF-α, IL-18 and 8-OHdG levels were measured by ELISA. Superoxide dismutase (SOD) activity was determined by spectrophotometry. RESULTS: Serum and urine TNF-α, IL-18 and 8-OHdG levels increased significantly with the degree of albuminuria, and were positively correlated with increased Ang-2. In contrast, SOD activity decreased with the degree of albuminuria and was negatively correlated with Ang-2. Multivariable linear regression analysis revealed that serum Ang-2 level was independently associated with serum levels of TNF-α (P<0.001), 8-OHdG (P=0.001), and IL-18 (P=0.003). Urinary Ang-2 level was independently associated with urinary TNF-α (P<0.001) and 8-OHdG (P=0.004) levels. CONCLUSION: TNF-α and 8-OHdG are associated with elevated urinary Angiopoietin-2 levels in type 2 diabetic patients with albuminuria.

Hydroxysafflor yellow A suppresses oleic acid-induced acute lung injury via protein kinase A.

Inflammation response and oxidative stress play important roles in acute lung injury (ALI). Activation of the cAMP/protein kinase A (PKA) signaling pathway may attenuate ALI by suppressing immune responses and inhibiting the generation of reactive oxygen species (ROS). Hydroxysafflor yellow A (HSYA) is a natural flavonoid compound that reduces oxidative stress and inflammatory cytokine-mediated damage. In this study, we examined whether HSYA could protect the lungs from oleic acid (OA)-induced injury, which was used to mimic ALI, and determined the role of the cAMP/PKA signaling pathway in this process. Arterial oxygen tension (PaO2), carbon dioxide tension, pH, and the PaO2/fraction of inspired oxygen ratio in the blood were detected using a blood gas analyzer. We measured wet/dry lung weight ratio and evaluated tissue morphology. The protein and inflammatory cytokine levels in the bronchoalveolar lavage fluid and serum were determined using enzyme-linked immunoassay. The activities of superoxide dismutase, glutathione peroxidase, PKA, and nicotinamide adenine dinucleotide phosphate oxidase, and the concentrations of cAMP and malondialdehyde in the lung tissue were detected using assay kits. Bcl-2, Bax, caspase 3, and p22(phox) levels in the lung tissue were analyzed using Western blotting. OA increased the inflammatory cytokine and ROS levels and caused lung dysfunction by decreasing cAMP synthesis, inhibiting PKA activity, stimulating caspase 3, and reducing the Bcl-2/Bax ratio. H-89 increased these effects. HSYA significantly increased the activities of antioxidant enzymes, inhibited the inflammatory response via cAMP/PKA pathway activation, and attenuated OA-induced lung injury. Our results show that the cAMP/PKA signaling pathway is required for the protective effect of HSYA against ALI.

SIRT6 regulates the proliferation and apoptosis of hepatocellular carcinoma via the ERK1/2 signaling pathway.

Hepatocellular carcinoma (HCC) is the most common type of liver cancer, and exhibits a high mortality rate. Sirtuin (SIRT)6 is a member of the sirtuin family, which may be useful targets in the treatment of tumors. The present study aimed to explore the expression of SIRT6 in numerous HCC cell lines and investigate the role of SIRT6 in the proliferation and apoptosis of the HCC cells, and the underlying mechanisms. Overexpression and silencing of SIRT6 were performed by transfection of Huh­7 cells with synthetic overexpression and small interfering RNA (siRNA) plasmids. Cell proliferation was evaluated using a Cell Counting Kit­8 assay. The apoptosis rate was measured via flow cytometry. Cloning efficiency was assessed using plate clone formation assays. The expression of mRNAs and proteins were determined via reverse transcription­quantitative PCR and western blot analyses, respectively. SIRT6 was overexpressed in Hep3B, Huh­7, MHCC­97H, MHCC­97L, MHCC­LM6, MHCC­LM3, YY­8103 and SK­hep­1 cell lines, compared with MIHA and HL­7702 normal liver cell lines. Overexpression of SIRT6 increased the proliferation of Huh­7 cells, upregulated the expression of Bcl­2 and phosphorylation of extracellular­signal regulated protein kinase (ERK), and decreased the expression of cleaved­caspase­3 and Bcl­2­associated X protein (Bax) in Huh­7 cells. siRNA­mediated silencing of SIRT6 decreased the proliferation and increased the apoptosis of Huh­7 cells, downregulated the expression of Bcl­2 and phosphorylated­ERK, and promoted the expression of cleaved­caspase­3 and Bax. The proliferation of Huh­7 cells was decreased using the ERK1/2 inhibitor U0126. The results suggested that SIRT6 affected the proliferation and apoptosis of HCC cells via the regulation of the ERK1/2 pathway, altering the activation of the intrinsic apoptosis pathway. SIRT6 may be a potential target for the treatment of HCC; however, its role requires further investigation.

Dehydrocostus lactone inhibits in vitro gastrinoma cancer cell growth through apoptosis induction, sub-G1 cell cycle arrest, DNA damage and loss of mitochondrial membrane potential.

INTRODUCTION: The purpose of the present study was to evaluate the antiproliferative activity of dehydrocostus lactone against human BON-1 cancer cell lines and to explore the possible underlying mechanism. MATERIAL AND METHODS: MTT cell viability assay was used to determine cytotoxic effects of dehydrocostus lactone in BON-1 cells. Fluorescence and transmission electron microscopic (TEM) techniques were used to study the effect of the compound on cellular morphology and apoptosis. Flow cytometry was used to assess the effect on cell cycle phase distribution. Effects of the drug on cell apoptosis and mitochondrial membrane potential were analyzed by flow cytometry using annexin v and rhodamine-123 as fluorescent probes. RESULTS: The results of the present study indicated that dehydrocostus lactone significantly (p < 0.01) inhibited the growth of BON-1 cancer cells. These growth inhibitory effects of dehydrocostus lactone on BON-1 were found to be time and concentration-dependent. The IC50 of dehydrocostus lactone were found to be 71.9 µM and 52.3 µM at 24 and 48 h time intervals respectively. The growth inhibitory effects of dehydrocostus lactone were found to be due to loss of mitochondrial membrane potential, the induction of apoptosis and sub-G1 cell cycle arrest. CONCLUSIONS: Dehydrocostus inhibits in vitro gastrinoma cancer cell growth and therefore may prove beneficial in the management of gastrinoma cancer.

Correlation between PPAR-α methylation level in peripheral blood and inflammatory factors of NAFLD patients with DM.

The correlation between the methylation levels of peroxisome proliferator-activated receptor-α (PPAR-α) in the peripheral blood and the inflammatory factors associated with non-alcoholic fatty liver disease (NAFLD) patients with diabetes mellitus (DM) was investigated. Thirty-two samples of normal liver tissues (group N) and 35 samples of liver tissues from NAFLD patients with DM (group M) were used for the present study. The levels of alanine transaminase (ALT) and aspartate transaminase (AST) were measured using commercially available kits. The accumulation of lipid droplets and glycogen in the two groups was determined through Oil Red O staining and Sudan III staining. mRNA expression of tumor necrosis factor-α (TNF-α), interleukin-lß (IL-1ß) and IL-6 in liver tissues of groups N and M were detected using reverse transcription-polymerase chain reaction (RT-PCR). In addition, western blotting was used to detect the protein expression of PPAR-α in liver tissues of both groups. The Statistical Product and Service Solutions (SPSS) 17.0 statistical software was used to analyze the expression difference of PPAR-α in liver tissues in the groups. The high levels of ALT and AST indicated severe liver injury in group M. Oil Red O staining and Sudan III staining showed a large number of lipid droplets and glycogen accumulation in the liver of group M patients. RT-PCR showed that the expression of inflammatory factors was extremely high and that the inflammatory injury was severe in the liver of group M patients. Western blotting showed that the expression of PPAR-α in group N was significantly higher than that in group M. ANOVA results showed that the expression of PPAR-α in liver tissues of groups N and M patients were statistically significantly different (P<0.01). Therefore, the abnormal expression of PPAR-α is closely associated with the occurrence and development of NAFLD complicated with DM, and that the abnormal expression of PPAR-α is closely related to inflammatory factors. Results from the present study suggest PPAR-α has important value in the study on NAFLD complicated with DM. The expression of PPAR-α can be used as a new basis for the diagnosis and treatment of NAFLD complicated with DM.

Correlation between PPAR-α methylation level in peripheral blood and atherosclerosis of NAFLD patients with DM.

We investigated the correlation between the methylation levels of peroxisome proliferator-activated receptor-α (PPAR-α) in the peripheral blood and atherosclerosis in patients with nonalcoholic fatty liver disease (NAFLD) with diabetes mellitus (DM). A total of 50 normal subjects (group N) and 50 NAFLD patients with DM (group M) were selected at Qilu Hospital of Shandong University. The levels of total cholesterol (TC), triglyceride (TG), high-density lipoprotein (HDL) and low-density lipoprotein (LDL) in groups N and M were detected. The mRNA and protein expressions of PPAR-α in groups N and M were detected using reverse transcription polymerase chain reaction (PCR) and immunofluorescence. The differences in expression of PPAR-α in group N and M and the correlation between PPAR-α methylation level and atherosclerosis were analyzed using SPSS17.0 statistical software. TC, TG, HDL and LDL in groups N and M were significantly different (P<0.01). Hematoxylin and eosin staining showed that the histopathological damage was severe in group M. PCR and immunofluorescence showed that PPAR-α was significantly higher in N than in group M (P<0.01). The abnormal expression of PPAR-α is closely related to atherosclerosis, indicating that the correlation between PPAR-α methylation levels in peripheral blood and atherosclerosis of NAFLD patients with DM can provide a new direction of diagnosis and treatment.

Promoter hypermethylation of death-associated protein kinase gene in cholangiocarcinoma.

BACKGROUND: Death-associated protein kinase (DAPK) is a Ca2+/calmodulin-regulated Ser/Thr kinase which is involved in apoptosis. The aberrant methylation of its promoter region CpG islands may be one of the important mechanisms of carcinogenesis. We studied the relationship of methylation status and expression of the DAPK gene with the clinical findings in cholangiocarcinoma. METHODS: Target DNA was modified by sodium bisulfite, coverting all unmethylated, but not methylated, cytosines to uracil, and subsequently detected by methylation-specific PCR. Moreover, mRNA expression of the DAPK gene was assessed by RT-PCR. RESULTS: Aberrant methylation of the DAPK gene was detected in 11 (30.6%) of 36 tissue specimens of cholangiocarcinoma, and in 2 (5.6%) of 36 specimens of adjacent normal tissues. DAPK mRNA was not expressed in tumor and adjacent tissues with hypermethylation of the DAPK promoter. There were no statistical differences in the extent of differentiation and invasion, lymph node metastasis or pathologic type between the methylated and unmethylated tissues. CONCLUSIONS: The frequency of DAPK gene methylation in cholangiocarcinoma is high and it may offer an effective means for earlier auxiliary diagnosis of the malignancy. The DAPK gene is probably suppressed by methylation, and it could become resistant to apoptosis and immunological surveillance. The DAPK gene epigenetically affected by methylation may be associated with the carcinogenesis of cholangiocarcinoma.

Thymoquinone induces cytotoxicity and reprogramming of EMT in gastric cancer cells by targeting PI3K/Akt/mTOR pathway.

Gastric cancer is one of the lethal causes of cancer-related deaths worldwide. The incidence and mortality rates of this disease is comparatively higher in China. In the current study, we evaluated the anticancer effects of Thymoquinone (TQ) against gastric cancer cells (MGC80-3 and SGC-7901) and normal noncancerous GES-1 cells and attempted to investigate the underlying mechanism. Our results indicated that TQ exhibited significant growth inhibitory effects on gastric cancer cells (MGC80-3 and SGC-7901). However, lower cytotoxicity was observed against normal GES-1 cells. Moreover, TQ could inhibit the colony formation potential of MGC80-3 and SGC-7901 cells in a dose-dependent manner. TQ also inhibited cell migration ability of the gastric cancer cells and down-regulated the expression of the mesenchymal genes such as N-cadherin, Vimentin, and TWIST. However, the epithelial markers such as E-cadherin and cytokeratin-19 were distinctly up-regulated in TQ-treated gastric cancer cells. Since PI3K/Akt/ mTOR plays an important role in progression and tumorigenesis, we also investigated the effect of TQ on PI3K/Akt/mTOR signalling pathway in gastric cancer cells. It was observed that TQ down-regulated the expression of some of the key proteins of this pathway. Taken together, we conclude that TQ may prove lead molecule for the treatment of gastric cancer.

Genetically lowered concentrations of circulating sRAGE might cause an increased risk of cancer: Meta-analysis using Mendelian randomization.

OBJECTIVES: To undertake a systematic meta-analysis of all variants in the gene encoding receptor for advanced glycation end products (RAGE) to summarize their associations with cancer risk and changes in the levels of circulating soluble RAGE (sRAGE), with the aim of determining possible causality between circulating sRAGE and cancer risk. METHODS: Articles written in English were retrieved from MEDLINE® and EMBASE® databases. Two researchers independently identified eligible articles and extracted the data (analysed using STATA® software version 12.0). RESULTS: Fifteen articles qualified for inclusion in the meta-analysis of the RAGE-cancer association and three examined the RAGE-sRAGE relationship. The 82Ser/82Ser genotype was significantly associated with overall cancer risk compared with the 82Gly/Gly genotype (odds ratio 1.75, 95% confidence interval [CI] 1.46, 2.10). Carriers of the 82Ser/82Ser genotype had significantly reduced circulating sRAGE concentrations compared with the 82Gly/82Gly genotype. Mendelian randomization analysis demonstrated that a reduction of 100, 200 and 300 pg/ml in circulating sRAGE concentrations was associated with a 1.11-fold (95% CI 1.06, 1.25), 1.24-fold (95% CI 1.11, 1.57) and 1.38-fold (95% CI 1.18, 1.96) increased risk of developing cancer, respectively. CONCLUSIONS: Genetically lowered concentrations of circulating sRAGE might cause an increased risk of cancer.

Clinical characteristics and surgical prognosis of hepatocellular carcinoma with bile duct invasion.

Objectives. Bile duct invasion (BDI) is a rare event in hepatocellular carcinoma (HCC). The present study aimed at investigating clinical characteristics and surgical outcome of HCC patients with bile duct invasion. Methods. 413 patients with HCC undergoing curative surgery were divided into two groups with (B(+)) and without BDI (B(-)). BDI was further classified as central type (B1) and peripheral type (B2). Survival was compared, and risk factors affecting prognosis were identified. Results. 35 (8.5%) patients were diagnosed BDI. Total bilirubin was significantly higher in B(+) group than in B(-) group (P < 0.001). Multiple lesions and large nodules (>5 cm) were predominantly identified in B(+) group (P < 0.01, resp.). Portal vein invasion was more frequently observed in B(+) than in B(-) group (P = 0.003). Univariate and multivariate analyses identified central BDI as a significant factor affecting prognosis of HCC patients (risk 1.3, 95% CI 1.1-2.2, P = 0.015). The gross overall survival of patients in B(+) was significantly worse than in B(-) (P = 0.001), which, however, was not different between B2 and B(-) (P > 0.05). Conclusions. Central but not peripheral BDI was associated with poorer prognosis of HCC patients. Curative surgical resection of tumors and invaded bile duct supplies the only hope for long-term survival of patients.

Hydroxysafflor yellow A of Carthamus tinctorius attenuates lung injury of aged rats exposed to gasoline engine exhaust by down-regulating platelet activation.

Long-term inhalation of gasoline engine exhaust (GEE) increases the risk of respiratory disease. Studies have suggested involvement of platelets in the development of some lung diseases. Hydroxysafflor yellow A (HSYA), a flavonoid compound, prevents hemostasis. Therefore, we investigated its effects on GEE-induced lung injury, and role of platelets in injury. Sixty-week-old male Sprague-Dawley rats were exposed to GEE for 4h/day for 6 weeks, and then grouped as follows: control, GEE, GEE+HSYA, GEE+HSYA+GW9662, and GEE+GW9662. Arterial oxygen tension (PaO2), carbon dioxide tension (PaCO2), pH, and the PaO2/fraction of inspired oxygen ratio (PaO2/FiO2) in the blood were detected using a blood gas analyzer. Wet/dry lung weight ratio, total protein in bronchoalveolar lavage fluid (BALF), and cytokine concentrations in serum and BALF were determined. Furthermore, cyclic adenosine monophosphate (cAMP) level and expression levels of target proteins were analyzed. Platelets were counted and their state was evaluated. HSYA attenuated GEE-mediated decreases in PaO2, PaO2/FiO2, platelet cAMP level, protein kinase A (PKA) activity, and peroxisome proliferator-activated receptor γ (PPARγ) expression. HSYA also attenuated GEE-mediated increases in lung permeability, cytokine levels in serum and BALF, plasma platelet count, and ADP-mediated platelet aggregation. Moreover, it suppressed GEE-induced increases in the expression of adhesion molecules and proinflammatory cytokines in platelets and lung tissue. Therefore, HSYA is therapeutically effective for GEE-mediated lung injury and acts by enhancing PKA activity and inhibiting platelet activation.

Epidemiological characteristics of thyroid nodules and risk factors for malignant nodules: a retrospective study from 6,304 surgical cases.

BACKGROUND: The prevalence of thyroid nodules (TN) is increasing rapidly. This study analyzed the epidemiological and clinical characteristics of TN in surgically treated patients and identified the risk factors for malignant nodules (MN) to provide more understanding of the differential diagnosis of TN. METHODS: A total of 6 304 TN cases who underwent thyroid surgery were included in this retrospective study. The clinical data were collected to evaluate the clinical and epidemiological characteristics and related risk factors for MN. The nature of TN (benign nodules (BN) or MN), medical records, laboratory data, and imaging data were analyzed. The risk factors for MN were screened using Spearman's rank correlation analysis and nonconditional binary Logistic regression analysis. RESULTS: The number of surgically treated TN cases increased yearly. A total of 34.33% of cases were MN and 65.67% were BN. Up to 56.74% of these cases underwent unnecessary surgery. Among the MN cases, papillary thyroid carcinoma accounted for 94%, in which 46.71% coexisted with benign thyroid disease and 32.28% with multiple foci. Single-related factor analysis showed that age, employment, disease duration, history of breast nodules and/or hypertension, the levels of serum thyroid-stimulating hormone (TSH), thyroglobulin antibody (TgAb), and thyroid peroxidase antibody (TPoAb), and ultrasound features of TN were related to MN. Stepwise nonconditional binary Logistic regression analysis showed that 13 factors may be the independent risk factors for MN, including <40 years old, previous history of breast nodules and/or hypertension, disease duration <1 month, employment, hypoechoic nodule, irregular nodules, nodule calcification, solid echo nodule, fuzzy boundary, rich blood flow within nodules, abnormal lymph nodes around the neck, nodule diameter <1 cm, and abnormally high TgAb. CONCLUSIONS: Our results demonstrate a rapid increase in surgically treated TN cases and ratio of MN and indicate unnecessary surgeries in some cases. This study also suggest that age, duration of thyroid disease, history of breast disease and/or hypertension, the levels of serum TSH, TgAb, and TPoAb, and ultrasound features of TN are related to MN, and some of these factors may be the risk factors for MN.

Effects of glucose and insulin on the H9c2 (2-1) cell proliferation may be mediated through regulating glucose transporter 4 expression.

BACKGROUND: The change of glucose transporter 4 (GLUT4) expression could influence glucose uptake in the myocardial cells and then effect myocardial metabolism, which maybe one of the factor for the diabetes cardiovascular disease. This study aimed to explore the influence of glucose and insulin at different concentrations on H9c2 (2-1) cell proliferation and its GLUT4 expression in vitro, and evaluate the correlation between myocardial cells proliferation and GLUT4 expression. This might be helpful for understanding the relationship between glucose metabolism and cardiovascular disease. METHODS: According to glucose concentrations in culture medium, cultured H9c2 rat myocardial cells were divided into five groups: control group (NC, glucose concentration 5.0 mmol/L), low glucose group (LG, glucose concentration 0.1 mmol/L), high glucose group 1 (HG1, glucose concentration 10 mmol/L), high glucose group 2 (HG2, glucose concentration 15 mmol/L), high glucose group 3 (HG3, glucose concentration 20 mmol/L). Then according to different insulin concentrations in culture medium, each group was further divided into two subgroups: normal insulin subgroup (INSc, insulin concentration 3.8 mU/L), high insulin subgroup (INSh, insulin concentration 7.6 mU/L). H9c2 (2-1) cells were cultured for 1, 2, 3 days, the proliferation of cells were assayed by cell counting Kit-8 assay, the expressions of GLUT4 mRNA and protein were detected with RT-PCR and Western Blotting technique, and the relation between myocardial cells proliferation and GLUT4 expression was evaluated. RESULTS: Compared with NC group, cell proliferation (OD value) was lower in LG, HG2, HG3 group but higher in HG1 group on the second and the third day (P < 0.05). There was a negative correlation between OD value and the glucose level in HG1, HG2, HG3 groups (P < 0.05). OD value in INSc subgroups was lower than that in INSh subgroups (P < 0.05). GLUT4 mRNA was lower in LG, HG2, HG3 groups than that in NC group (P < 0.05). Compared with NC group, GLUT4 mRNA level in HG1 group was higher on the first day but lower on the second and third day (P < 0.05). In HG1, HG2 and HG3 groups, GLUT4 mRNA level had a negative correlation with the level of glucose (P < 0.05). GLUT4 mRNA in INSc subgroups was lower than that in INSh subgroups (P < 0.05). The expression of GLUT4 protein was similar to that of GLUT4 mRNA. There was a positive correlation between H9c2 cell proliferation and GLUT4 expression (P < 0.02). CONCLUSIONS: Glucose levels could regulate glucose uptake in myocardial cells through influencing GLUT4 expression, and thus affected the cell proliferation and cell function. Insulin levels could affect the myocardial cell function by regulating GLUT4 expression. Effects of glucose and insulin on the myocardial cells proliferation might be mediated through regulating GLUT4 expression. There may be a mechanism of hyperglycemia pre-accommodation (HGPA) in myocardial cells mediated through regulation of GLUT4 expression.

Basal insulin therapy strategy is superior to premixed insulin therapy in the perioperative period blood glucose management.

BACKGROUND: The probability and risk of operations increase in patients with type 2 diabetes mellitus. For diabetic patients, blood glucose control is a key factor to improving the prognosis of surgery. During perioperative period, insulin therapy is usually advised to be used for surgical patients with type 2 diabetes. However, the insulin regimen which one is better remains controversial. In this study, we estimated the efficacy, safety and advantage of different insulin therapy strategy during perioperative period. METHODS: A total of 1086 cases of surgical patients with type 2 diabetes mellitus enrolled in the present study. According to the glucose level at admission, all patients were divided into relatively high glucose group (group A, fasting blood glucose (FBG) ≤13.9 mmol/L) and higher glucose group (group B, FBG >13.9 mmol/L). Patients in group A randomly accepted premixed insulin twice a day, or basal insulin plus oral medications, and were divided into group A1 and A2 respectively. Patients in group B randomly received premixed insulin twice daily, basal insulin plus oral hypoglycemic agents, or basal insulin plus preprandial insulin, and were divided into group B1, B2 and B3 respectively. The data of the preoperative preparation time, the daily doses of insulin used in different periods, postoperative incision healed installments, hypoglycemic events, the total hospitalization time, postoperative complications were all collected and statistically analyzed. RESULTS: Compared the main outcome measures in groups treated by premixed insulin therapy, both in preoperative preparation and postoperative period, the daily insulin dosage and the frequency of hypoglycemic events were decreased in groups treated by basal insulin therapy (P < 0.05). The preoperative preparation time and the total hospitalization time in groups with basal insulin therapy were shorter than that in groups with premixed insulin therapy (P < 0.05). The incision healing rate of stage I, II and III among different therapy protocols were significantly different (P < 0.05). CONCLUSIONS: Basal insulin therapy could be used in diabetic patients undergoing elective major and medium surgery during whole perioperative period. Basal insulin therapy strategy, including a single injection of basal insulin and basal insulin plus preprandial insulin injection subcutaneously, is superior to premixed insulin therapy in the perioperative blood glucose management, and it could be viewed as the best choice in glucose control during perioperative period.

Chemerin and apelin are positively correlated with inflammation in obese type 2 diabetic patients.

BACKGROUND: As two novel adipocytokines, chemerin and apelin play a key role in the pathological process of insulin resistance (IR), glucose metabolism and obesity, researchers have found that the levels of chemerin and apelin changed significantly in type 2 diabetic patients with obesity, however, the underlying mechanism involved remains unclear. The aim of this study was to investigate whether chemerin and apelin play an important role in the pathophysiologic proceeding of diabetes. METHODS: This study enrolled 81 newly diagnosed obese type 2 diabetes mellitus (T2DM) patients (T2DM group, n = 81). All the patients were randomly assigned to DM1 group treated with metformin (n = 41) and DM2 group treated with pioglitazone (n = 40). After hypoglycemic agents treatment, patients under better blood glucose control were chosen to be given antioxidant treatment. Another 79 subjects without T2DM were recruited as normal control group (NC group), including 40 subjects (NC1 group) with normal body mass index (BMI) and 39 obese subjects (NC2 group). Levels of chemerin, apelin, BMI, tumor necrosis factor-α (TNF-α), homeostasis model assessment of IR (HOMA-IR) and 8-isoprotaglandim F2α (8-iso-PGF2α) were examined at baseline and post-treatment. The relationship between chemerin, apelin and BMI, TNF-α, HOMA-IR, 8-iso-PGF2α was analyzed. RESULTS: The baseline levels of chemerin, apelin, TNF-α, HOMA-IR and 8-iso-PGF2α in T2DM group were significantly higher than normal control group (P < 0.001). All indices mentioned above were significantly decreased after treatment (P < 0.05). In T2DM patients treated with pioglitazone, indices mentioned above except for HOMA-IR, were decreased significantly compared with patients treated with metformin (P < 0.05). After antioxidant treatment using lipoic acid, levels of chemerin, apelin, TNF-α and 8-iso-PGF2α were further significantly decreased (P < 0.05). Correlation analysis showed that the levels of chemerin and apelin correlated positively with BMI, TNF-α, HOMA-IR and 8-iso-PGF2α before and after treatment with hypoglycemic agents (P < 0.01). The levels of chemerin and apelin also had positive correlation with TNF-α and 8-iso-PGF2α after antioxidant treatment (P < 0.05). CONCLUSIONS: The levels of chemerin and apelin in obese T2DM patients are closely related to IR. The increased levels may be a result of compensatory response to IR, and also may be the causative factor of IR. The levels of chemerin and apelin correlate closely with oxidative stress and inflammation. The two adipokines may be inflammatory factors playing important roles in the initiation and development of obese T2DM. Chemerin and apelin are related to the pathophysiology of IR, oxidative stress and inflammation.

[Inhibitory effects of high mobility group box 1 antisense nucleotide on invasion of human pancreatic cancer cell line PCNA-1].

BACKGROUND & OBJECTIVE: High mobility group box 1 (HMGB1) is abundantly expressed in most of immature, and malignant cells, and is closely related to cell proliferation, invasion, and migration. We have previously proved HMGB1 over-expressed in pancreatic cancer tissues. This study was to evaluate the in vitro effects of HMGB1 antisense nucleotide on human pancreatic cancer cells invasion, and explore the underlying mechanism. METHODS: An eukaryotic expression vector containing antisense-HMGB1 was transfected into human pancreatic cancer cell line PCNA-1. The expression of HMGB1, matrix metalloproteinase-2 (MMP-2),and MMP-9 mRNA before and after transfection was detected by reverse transcriptase-polymerase chain reaction(RT-PCR); HMGB1 protein expression was examined by Western blot. The activities of MMP-2, and MMP-9 were analyzed by gelatin zymography. The in vitro invasive ability of PCNA-1 cells was determined by Boyden chambers method. RESULTS: Antisense-HMGB1 eukaryotic expression vector was successfully constructed. HMGB1 mRNA expression in PCNA-1 cells was effectively inhibited by antisense-HMGB1. HMGB1 protein expression was also suppressed upon transfecting (P< 0.05). Antisense-HMGB1 transfected cells showed lower MMP-2 and MMP-9 mRNA expression (P< 0.01). MMP-2 and MMP-9 activities were also inhibited by antisens-HMGB1 (P< 0.01). Cells migrating through the Boyden chamber membrane was significantly reduced as compared with the control (P< 0.01). CONCLUSIONS: HMGB1 plays a crucial role in the invasion of pancreatic cancer, and blocking HMGB1 may be a potential strategy in preventing the migration of pancreatic cancer.