Hyperglycemia has been identified as one of the important factors involved in the microvascular complications of diabetes, and has been related to increased cardiovascular mortality. Endothelial damage and dysfunction result from diabetes; therefore, the aim of this study was to determine the response of endothelial cells to stressful stimuli, modelled in normal and high glucose concentrations in vitro. Eahy 926 endothelial cells were cultured in 5 mmol/L or 30 mmol/L glucose conditions for a 24 hour period and oxidative stress was induced by exposure to hydrogen peroxide (H2O2) or tumour necrosis factor- α (TNF- α ), following which the protective effect of the glucocorticoid dexamethasone was assessed. Apoptosis, necrosis and cell viability were determined using an ELISA for DNA fragmentation, an enzymatic lactate dehydrogenase assay and an MTT assay, respectively. High glucose significantly increased the susceptibility of Eahy 926 cells to apoptosis in the presence of 500 μmol/L H2O2, above that induced in normal glucose (P<0.02). A reduction of H2O2- and TNF- α -induced apoptosis occurred in both high and low glucose after treatment with dexametha-sone (P<0.05). Conclusion high glucose is effective in significantly augmenting stress caused by H2O2, but not in causing stress alone. These findings suggest a mechanism by which short term hyperglycemia may facilitate and augment endothelial damage.

Objective To study the protective effect of fluvastatin,one of the HMG-CoA reductase inhibitors (statins),against oxygen radical-induced oxidative damages in human aortic endothelial cell,and the role of Bcl-2 in this protection.Methods Human aortic endothelial cells with or without Bcl-2 siRNA transfection were subjected to 1-100 nM of fluvastatin and 100 la hydrogen peroxide for 24 hours.Bcl-2 mRNA and protein expression were measured by Taqman quantitative PCR and Western blotting.Cell apoptosis was measured by normal and fluorescent microscopy and Cell Death Detection ELISA.Results In the Bcl-2-expressed cells,fluvastatin significantly reversed hydrogen peroxide-induced microscopic apoptosis and apoptotic DNA fragmentation,which were accompanied by a markedly upregulation of Bcl-2 expression by fluvastatin.However,the endothelial protection by fluvastatin was completely lost in Bcl-2 siRNA transfected cells.Conclusion Fluvastatin protects human endothelial cells against oxygen radical-induced cell apoptosis in vitro,and this protection seemed to be mediated in a Bcl-2 dependent pathway.(J Geriatr Cardil 12008;5:33-38)

Objective The aim of this study was to determine if isoflavone genistien has protective effects against high glucose-induced cell apoptosis in human aortic endlthelial cells,and investigate the possible mechanism for this protection.Methods Human aortic endothelial cells subjected to normal (5mmol/L) or high glucose (25mmol/L) were treated with genistein at 0,50,100nmol/L.Parallel experiments were performed with 100nM 17b-estradiol,and also in the presence and absence of the pure anti-estrogen ICI-182,780 (100nmol/L).The effects on cell apoptotic DNA fragmentation were determined using cell death ELISA,and the effects on cellular proliferation were determined using tritiated thymidine incorporation assay.Estrogen receptor expression was detected by Taqman quantitative PCR.Results Genistein at 100nmol/L significantly reduced high glucose-induced DNA fragmentation,and reversed cell DNA synthesis inhibition (P＜0.001) after 24 hours' incubation.The effect of genistein was completely blocked by ICI-182,780administration.Estrogen receptor beta,but not alpha was found to be expressed in these cells.Conclusion Isoflavone genistein shows protection against high glucose-induced cell damage through estrogen receptor beta,reducing apoptotic DNA damage and protecting from the inhibition of cell proliferation.

The occurrence of malignant tunors is increasing annually around the world.In every year,8.2 million patients died of malignant tumors in which about 90％ patients died of malignant tumors associated multiple organs metastases.Inhibiting tumor metastasis is the key event to prolong the survival time of patients.With the further research,tumor-vessel associated extravascular tumor metastasis is playing an increasingly important role in the clinical application.This paper summarizes the new developments of tumor-vessel associated extravascular tumor metastasis to provide possible ideas for the therapy of malignant tumors.

Objective To investigate the anatomic structure of the Chinese people,develop the procedure of minimally invasive total hip arthroplasty through the anterolateral intermuscular approach and investigate its clinical outcome.Methods Three fresh adult cadavers(6 hips)were used for study of the anatomic construction of the anterolateral intermuscular approach in Chinese people.Sixteen patients were treated with minimally invasive total hip arthroplasty through anterolateral intermuscular approach.The clinical results and operation technique were recorded.Results The anterolateral intermuscular approach was a triangle muscular interval slightly parallel to the femur.The medial-superior angle of the triangle muscular interval consists of the anterior border of gluteus medius and tensor fascia lata muscle with juncture of muscles,where the inferior branch of superior gluteal nerve entered into tensor fascia lata muscle.The average incision length was 8.8 cm(7-10 cm),with mean blood loss of 350 ml(250-550 ml).The patients took out-of-bed activity 3-5 days after operations.During operations,anterior border injury in deep portion of the gluteus medius muscle was observed in seven patients and the injured muscles were trimed or repaired.All patients were followed up for 18-39 months(averaged 27.7 months).Most of the patients had excellent location of the phantoms,except that one acetabulum had a little pitch angle and two acetabulums had a little abduction angle.No complication was observed.The mean Harris scores of hip for all patients was increased from preoperative(39.1±6.7)points to(80.6±11.3)points on six month,(88.7±9.6)points on 12 month and(91.4±13.5)points on 24 months(11 patients).No patient suffered from gluteus medius muscle weakness during the follow-up.ConclusionAnterolateral intermuscular approach has the advantages of simple anatomic construction,small incision,little operative injury,muscle sparing and fast recovery without separate muscle or tendon and is suitable for the Chinese patients.Exact incision and special operative instruments should be emphasized to avoid the increase of acetabular pitch angle and abduction angle.

Objective To provide immunological evidence for clinical transfer of chemical extracted acellular nerve allografL Methods One hundred and twenty-eight BALB/C mice were randomly divided into 4 groups of equal size according to their different treatments:negative contrast group(NC),fresh autograft group(AG),fresh allogeneic nerve group(FN)and chemical extracted aceflular allogeneic nerve group(CEN).Then we implanted various kinds of nerve grafts into the thigh muscle of BALB/C mice in corresponding groups.At 3,7,14,28 days postoperatively,8 mice from each group were killed each time to harvest their spleens,from which T lymphocytes were collected.Theu monoclonal antibodies(CD3,CD4 CD8 CD25,IL-2,IFN-γ, TNF-α)were added into the suspension.Then fluorescence.activated cell sorting(FACS)was used to determine the positive rates of cells combined with the above monoclonal antibodies. Results There were no statistically significant differences between CEN group,NC group,and AG group,but indexes of FN group were significantly higher than those of the other 3 groups at corresponding time points. Conclusion There is no obvious immune reiection of chemical extracted acellular nerve allograft when compared with fresh nerve autograft.

Secondary immune response is an important endogenous mechanism of neurological injury after ischemic stroke.Spleen and interferon-γγplay an important role in it.Monitoring the spleen size and the level of interferon-γγhave an important reference significance for the severity of stroke and outcome assessment.

Objective To study the impact of different insulin levels on the conversion from impaired glucose tolerance (IGT) to type 2 diabetes mellitus (T2DM),through analysis of different glycometabolism condition among quinquagenarian population.Methods Subjects enrolled were Beijing habitants who received annual physical examination [ including oral glucose tolerance test (OGTI) ] in the Chinese PLA General Hospital from 2005-2007.According to the OGTT results,the subjects were divided into three groups,including normal glucose tolerance-non-hyperinsulinemia group (NGT-NHIns),IGT-hyperinsulinemia group (IGT-Hins) and IGT-non-hyperinsulinemia group (IGT-NHINS).The prognosis between the year 2009 and 2010 of the three groups was observed.Hyperinsulinemia was diagnosed with fasting serum insulin ≥ 15 mU/L and/or 2-hour serum insulin ≥ 80 mU/L after glucose loading.Results The rate of case number of conversion to T2DM in IGT-NHIns group (42/133) was higher than that in IGT-Hins group (24/154) or NGT-NHIns group (12/126).The HOMA insulin resistance index (HOMAIR) of individuals with IGT-NHIns was lower than that of IGT-Hins [ 0.96 (0.40,3.53 ) vs 2.04 (0.59,23.20),P ＜ 0.05 ],while whole body insulin sensitivity index (WBISI) was higher than that of IGT-Hins [ 7.48 (3.20,31.35 ) vs 3.28 ( 0.86,7.67 ),P ＜ 0.05 ].Modified β-cell function index ( MBCI ) and insulin secretion index (ISI) in IGT-NHIns was poorer than that of IGT-Hins respectively [ 2.57 (0.58,10.98) vs5.17(1.04,65.09); 7.66 (0.99,28.40) vs 17.56 (4.18,96.46),allPvalues ＜0.01].Conclusions The risk of IGT-NHIns progressing into T2DM is higher than that of IGT-Hins. For the prevention of T2DM,individuals with IGT-NHIns should be paid more attention than keeping an eye on IGT-Hins patients.Early control of risk factors could protect β cell function and prevent the progression to T2DM.

This study inquired into the mechanisms of co-immobilized cytokines and free cytokines-induced apoptosis on HeLa cells. With the use of photochemical fixed method, TNF-alpha/IFN-gamma were co-immobilized on a 24-well polystyrene culture plate. HeLa cells were stained with fluorescent probe JC-1 to detect the changes of mitochondrial membrane potential (deltapsim), and then were examined by flow cytometry. The results showed that co-immobilized cytokines could induce the apoptosis of HeLa cells in a dose-independent manner. When treated with low-dose of co-immobilized cytokines (20ng/ml), the mitochondrial membrane potential (deltapsim) of HeLa cells continually decreased in 6 days. These indicate that low dose co-immobilized cytokines have a long-term of apoptosis-inducing effect on HeLa cells. We assume that there is close relationship between the mitochondrial membrane potential decrease and the apoptosis of HeLa cells.
Apoptosis ; drug effects ; Dose-Response Relationship, Drug ; HeLa Cells ; Humans ; Immobilized Proteins ; pharmacology ; Interferon-gamma ; pharmacology ; Membrane Potential, Mitochondrial ; drug effects ; Mitochondrial Membranes ; drug effects ; physiology ; Tumor Necrosis Factor-alpha ; pharmacology

ObjectiveTo investigate the clinical features and gene mutation characteristics of neonatal intrahepatic cholestasis caused citrin deficiency (NICCD) in northern China. MethodsA total of 23 pediatric patients in northern China who were diagnosed with NICCD by blood tandem mass spectrometry and/or gene detection in Department of Gastroenterology, Children’s Hospital Affiliated to Capital Institute of Pediatrics, from January 2015 to December 2018 were enrolled as NICCD group, and 36 pediatric patients with idiopathic neonatal cholestasis (INC) who had unclarified etiology after a series of examinations during the same period of time were enrolled as INC group. A retrospective analysis was performed for the clinical manifestation, laboratory examination, pathology, blood/urine metabolic screening, and gene sequencing results of the pediatric patients in the NICCD group, and follow-up was performed to observe their outcome; biochemical parameters were compared between the two groups. The independent samples t-test was used for comparison of normally distributed continuous data, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data; the chi-square test was used for comparison of categorical data between groups. ResultsAmong the 23 patients in the NICCD group, 10 had hypoglycemia, 13 had hypoalbuminemia, 17 had hyperammonemia, and 15 had hyperlactacidemia; 15 had an increase in low-density lipoprotein, 6 had an increase in cholesterol, and 7 had an increase in triglyceride; 17 had prolonged prothrombin time, and 16 had prolonged activated partial thromboplastin time (APTT). Compared with the INC group, the NICCD group had significantly higher gamma-glutamyl transpeptidase (GGT), total bile acid (TBA), and APTT and a significantly lower albumin (Alb) level (Z=-2.487, Z=-3.528, t=3.532, t=-2.24, all P＜0.05). For the patients with NICCD, blood tandem mass spectrometry showed that the most common abnormalities were the increased levels of arginine, citrulline, methionine, free carnitine, and long-chain acylcarnitine, while urinary gas chromatography showed the increased levels of 4-hydroxyphenyllactic acid, galactose, galactitol, and galactonic acid. Gene detection was performed for all 23 patients and identified 16 pathogenic mutations, among which 7 were newly discovered, namely ivs14-9a＞G, c1640 G＞A, c.762T＞A, c.736delG, c.1098 T del, c.851G＞A, and c.550G＞A. Except for the 2 patients who were lost to follow-up, the levels of aminotransferases and bilirubin gradually returned to normal in 21 patients after 2-6 months of treatment; none of them showed delayed growth and development after being followed up to the age of 1 year, and 2 of them developed dietary preference (they liked fish and meat and did not like staple food). ConclusionAbnormalities of blood GGT, TBA, Alb, and APTT may provide ideas for the differential diagnosis of NICCD and INC. NICCD gene mutations in northern China are heterogeneous and most patients tend to have a good prognosis.