Effect of oxymatrine on liver gluconeogenesis is associated with the regulation of PEPCK and G6Pase expression and AKT phosphorylation.

An increase in liver gluconeogenesis is an important pathological phenomenon in type 2 diabetes mellitus (T2DM) and oxymatrine is an effective natural drug used for T2DM treatment. The present study aimed to explore the effect of oxymatrine on gluconeogenesis and elucidate the underlying mechanism. Male Sprague-Dawley rats were treated with a high-fat diet and streptozotocin for 4 weeks to induce T2DM, and HepG2 cells were treated with 55 mM glucose to simulate T2DM in vitro. T2DM rats were treated with oxymatrine (10 or 20 mg/kg weight) or metformin for 4 weeks, and HepG2 cells were treated with oxymatrine (0.1 or 1 µM), metformin (0.1 µM), or oxymatrine combined with MK-2206 (AKT inhibitor) for 24 h. Fasting blood glucose and insulin sensitivity of rats were measured to evaluate insulin resistance. Glucose production and uptake ability were measured to evaluate gluconeogenesis in HepG2 cells, and the expression of related genes was detected to explore the molecular mechanism. Additionally, the body weight, liver weight and liver index were measured and hematoxylin and eosin staining was performed to evaluate the effects of the disease. The fasting glucose levels of T2DM rats was 16.5 mmol/l, whereas in the control rats, it was 6.1 mmol/l. Decreased insulin sensitivity (K-value, 0.2), body weight loss (weight, 300 g), liver weight gain, liver index increase (value, 48) and morphological changes were observed in T2DM rats, accompanied by reduced AKT phosphorylation, and upregulated expression of phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6Pase). High-glucose treatment significantly increased glucose production and decreased glucose uptake in HepG2 cells, concomitant with a decrease in AKT phosphorylation and increase of PEPCK and G6Pase expression. In vivo, oxymatrine dose-dependently increased the sensitivity of T2DM rats to insulin, increased AKT phosphorylation and decreased PEPCK and G6Pase expression in the liver, and reversed the liver morphological changes. In vitro, oxymatrine dose-dependently increased AKT phosphorylation and glucose uptake of HepG2 cells subjected to high-glucose treatment, which was accompanied by inhibition of the expression of the gluconeogenesis-related genes, PEPCK and G6Pase. MK-2206 significantly inhibited the protective effects of oxymatrine in high-glucose-treated cells. These data indicated that oxymatrine can effectively prevent insulin resistance and gluconeogenesis, and its mechanism may be at least partly associated with the regulation of PEPCK and G6Pase expression and AKT phosphorylation in the liver.

Low expression of miR­532­3p contributes to cerebral ischemia/reperfusion oxidative stress injury by directly targeting NOX2.

NADPH oxidase 2 (NOX2) is a major subtype of NOX and is responsible for the generation of reactive oxygen species (ROS) in brain tissues. MicroRNAs (miRNAs/miRs) are important epigenetic regulators of NOX2. The present study aimed to identify the role of NOX2 miRNA­targets in ischemic stroke (IS). A rat cerebral ischemia/reperfusion (CI/R) injury model and a SH­SY5Y cell hypoxia/reoxygenation (H/R) model were used to simulate IS. Gene expression levels, ROS production and apoptosis in tissue or cells were determined, and bioinformatic analysis was conducted for target prediction of miRNA. In vitro experiments, including function­gain and luciferase activity assays, were also performed to assess the roles of miRNAs. The results indicated that NOX2 was significantly increased in brain tissues subjected to I/R and in SH­SY5Y cells subjected to H/R, while the expression of miR­532­3p (putative target of NOX2) was significantly decreased in brain tissues and plasma. Overexpression of miR­532­3p significantly suppressed NOX2 expression and ROS generation in SH­SY5Y cells subjected to H/R, as well as reduced the relative luciferase activity of cells transfected with a reporter gene plasmid. Collectively, these data indicated that miR­532­3p may be a target of NOX2 and a biomarker for CI/R injury. Thus, the present study may provide a novel target for drug development and IS therapy.

Right ventricular outflow tract septal pacing versus apical pacing: A prospective, randomized, single-blind 5-years follow-up study of ventricular lead performance and safety / 华中科技大学学报（医学）（英德文版）

Lead placement for ventricular pacing variably impacts the physiological benefit of the patient. This study evaluated the ventricular lead performance and safety of right ventricular outflow tract septal pacing in patients with bradyarrhythmia in South China over 60-month follow-up. Totally, 192 patients (108 males, and 84 females, 63±21 years old) with bradyarrhythmia were randomly divided into two groups. The right ventricular outflow tract septum (RVOTs) group had lead placement near the septum (n=97), while the right ventricular apex (RVA) group had a traditional apical placement (n=95). RV septal lead positioning was achieved with a specialized stylet and confirmed using fluoroscopic projection. All patients were followed up for 60 months. Follow-up assessment included stimulation threshold, R-wave sensing, lead impedance and lead complications. The time of electrode implantation in both the ROVTs and RVA groups were significantly different (4.29±0.61 vs. 2.16±0.22 min; P=0.009). No differences were identified in threshold, impedance or R-wave sensing between the two groups at 1st, 12th, 36th and 60th month during the follow-up period. No occurrence of electrode displacement, increased pacing threshold or inadequate sensing was found. The long-term active fixation ventricular electrode performance in RVOTs group was similar to that in RVA group. RVOTs pacing near the septum using active fixation electrodes may provide stability during long-term follow-up period.

Correlation of angiotensin-converting enzyme 2 gene polymorphisms to essential hypertension and ischemic stroke / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the relationship between angiotensin-converting enzyme 2 (ACE2) gene polymorphisms and the risk factor for essential hypertension (EH) with concurrent ischemic stroke in southern Chinese population.</p><p><b>METHODS</b>The G9570A polymorphism in ACE2 gene were detected in 139 patients with EH and stroke using polymerase chain reaction-restriction fragment length polymorphism. Detailed clinical and biochemistrical data of the patients, including the pulse pressure, high sensitivity C-reactive protein (hsCRP), intima-media thickness (IMT), high-density lipoprotein cholesterol (HDL-C) and uric acid levels, were collected to study the relationship between ACE2 gene and the risk factor of EH and stroke.</p><p><b>RESULTS</b>The levels of hsCRP (OR=1.022), uric acid (OR=1.224), IMT and pulse pressure was positively correlated to the incidence of EH and stroke. The pulse pressure, hsCRP, IMT, and HDL-C levels in male stroke patients carrying A allele was significantly higher than those in patients carrying G allele (P<0.05). In female stroke patients, the pulse pressure, hsCRP, IMT, and HDL-C levels were also significantly different with regard to the genotype of ACE2 gene (P<0.05).</p><p><b>CONCLUSIONS</b>The patients with EH and ischemic stroke carrying the A/AA allele of ACE2 gene have higher risks than those carrying other allele, and can be also more vulnerable to stroke recurrence.</p>

Relationship of angiotensin-converting enzyme 2 gene polymorphism with the prognosis of hypertensive stroke patients / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the relationship between angiotensin-converting enzyme 2 (ACE2) gene G9570A polymorphisms and the clinical outcome of stroke patients with essential hypertension (EH) in South China Han population.</p><p><b>METHOD</b>The ACE2 gene polymorphisms were detected in 141 stroke patients with EH and 156 patients with EH using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The genetic marker was tested for its association with the baseline measurements and clinical outcomes of the patients over a median follow-up period of 22 months. As the ACE2 gene is X-linked, analyses were performed for male and female patients separately.</p><p><b>RESULTS</b>The A allele frequency in the stroke patients was significantly different from that in the EH patients, and the AA allele frequency in the female patients was significantly different between the two groups (P<0.01). Kaplan-Meier model analysis showed that ACE2 gene polymorphism was not associated with the the patients' prognosis (P>0.05). Multivariate Cox's proportional hazard regression model identified age (RR=1.057, 95%CI: 1.020, 1.095), blood glucose (RR=1.575, 95%CI: 1.178, 2.104), hypertriglyceridemia (RR=1.947, 95%CI: 1.503, 2.780), blood creatinine (RR=1.034, 95%CI: 1.001, 1.068), and blood uric acid (RR=1.056, 95%CI: 1.002, 1.097) as the risk factors associated with the mortality.</p><p><b>CONCLUSION</b>Stroke occurs more likely in hypertensive patients carrying the A/AA allele than those carrying other alleles. The ACE2 gene G9570A polymorphisms may be associated with the occurrence of stroke in EH patients in South China, but may not have a strong correlation to the prognosis.</p>

Laparoscopy-assisted D2 total gastrectomy in advanced gastric cancer / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the efficacy and advantages of laparoscopy-assisted total gastrectomy (LATG) with D2 dissection of lymph nodes versus conventional open D2 total gastrectomy (OTG) in advanced gastric cancer.</p><p><b>METHODS</b>One hundred and twenty-five patients with advanced gastric cancer in the middle or upper third of the stomach were operated on from July 2005 to March 2007. Of the patients, 59 cases received LATG and 66 OTG with D2 lymph nodes dissection. Clinical data were recorded and compared between the two groups.</p><p><b>RESULTS</b>No patient in the LATG group converted to conventional operation with laparotomy. No operation mortality and no severe morbidity occurred in LATG group. As compared with OTG group, in LATG group operation time was longer [(330 +/- 71) min vs. (261 +/- 54) min, P =0.005] in LATG group, but with similar number of lymph node retrieval (36 +/- 13 vs. 34 +/- 16, P =0.450), less operation blood loss [(175 +/- 101) ml vs. (359 +/- 210) ml, P =0.003], earlier recovery of bowel activity (P = 0.015), and a shorter duration of fever after operation (P = 0.024).</p><p><b>CONCLUSIONS</b>LATG with D2 lymph node dissection in advanced gastric cancer is safe and technically feasible with better operative access and visual field, less operation blood loss and earlier recovery.</p>

Construction of rat bowel transforming growth factor beta1 recombinant adenovirus vector / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To construct a recombinant adenovirus vector which regulates the expression of rat transforming growth factor beta (TGF-beta1).</p><p><b>METHODS</b>Total RNA was extracted from F344/N rat small intestine pre-treated with Con A to clone TGF-beta1. pTRE-shuttle vector was used as mediator to ligate TGF-beta1 gene and backbone of replication-incompetent adenoviral vector. The constructed recombinant adenovirus contained tetracycline-responsive element which could regulate the expression of inserted genes. After identification, the desired recombinant adenovirus was packaged in HEK 293 cells. Supernatant of high titer adenovirus was collected to detect the TGF-beta1 gene expression by green fluorescent protein(GFP).</p><p><b>RESULTS</b>The constructed recombinant adenovirus was identified by restriction endonucleases cutting, sequencing, PCR and GFP examination.</p><p><b>CONCLUSION</b>Rat TGF-beta1 recombinant adenovirus is established successfully, which provides material and evidence for further research of dendritic cell (DC) modified by TGF-beta1 to induce immune tolerance in rat heterotopic small bowel transplantation.</p>

Relationship Between the Angiotensin-Converting Enzyme 2 Genotype and Effect of Irbesartan on Left Ventricular Structure and Function in Hypertensive Patients / 中华高血压杂志

Background Experimental data have shown tnat polymorpnisms in tne angiotensm-converting en- zyme 2(ACE2)gene are related to echocardiographically determined parameters of left ventricular mass,structure or function in the general population whether ACE2 genotype influences the effect of angi0tensin Ⅱ receptor blocker which improve left ventricular remodeling and function is unknown.Objective To investigate the association be- tween ACE2 gene G9570A polymorphism and the effect of irbesartan on left ventricular structure and function in hy- pertensive patients.Methods Two hundred and five male patients and 190 female patients who were preliminaryly diagnosised with mild and moderate essential hypertension were treated with irbesartan for 48 weeks with initial dose of 150 mg/d and titrated to 300 mg/d to reach the targed BP.Gene polymorphisms of ACE2 G9570A were detected by PCR-RFLP methods.The association between changes in the SBP,DBP,parameters of left ventricular struc- ture and function and genotypes of the ACE2 gene locus were analyzed.Results Irbesartan reducted in blood pres- sure in all patients(P

Expression of mucosal addressin cell adhesion molecule-1 during small bowel graft rejection in rat / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the expression of mucosal addressin cell adhesion molecule-1(MAdCAM-1) during small bowel graft rejection and the effects of MAdCAM-1 on the development of acute rejection.</p><p><b>METHODS</b>Rat heterotopic small bowel transplantation (SBT) was performed in F344/N rats with syngeneic and allogeneic (BN-F344/N) grafts. Bowel and gut-associated lymphoid tissue(GALT) samples were collected from small bowel transplants on postoperative day(POD) 1, 3, 5 and 7. Histopathology assessment of the grafts was conducted to identify the rejection. MAdCAM-1 was detected by immunohistochemistry and Western blot.</p><p><b>RESULTS</b>During acute rejection, MAdCAM-1 was highly-expressed on gut lamina propia and GALTs, particularly on vascular endothelial cells in the gut lamina propia. There were no change of MAdCAM-1 expression in syngeneic grafts from POD1 to POD7. In allogeneic grafts, MAdCAM-1 expression in mesenteric lymph nodes was down-regulated, while up-regulated on the vascular endothelial cells in the lamina propria during acute rejection.</p><p><b>CONCLUSION</b>Alteration of MAdCAM-1 expression may be associated with the development of SBT graft rejection.</p>