BACKGROUND: Research has proved that nifedipine can improve the functions of vascular endothelial cells. However, it still needs further study on its mechanism on dilating vessels.OBJECTIVE: To observe the effects of controlled-release nifedipine to nitrogen monoxide(NO) and induced nitric oxide synthase(iNOS) in spontaneously hypertensive rats(SHR).DESIGN: Comparative observational study based on experimental animals.SETTING: Experimental animal centre in a medical college.MATERIALS: This study was completed in Experimental Animal Centre of Shandong University during April to May 2002. Twenty-one recombinant inbred rats of clean class with body mass of(300 ± 2) grams were provided by Experimental Animal Centre of Shandong University. They were randomly divided into three groups: control group, normal group and low dose group,each with 7 rats.METHODS: Saline(10 mL/kg), 10 mL/kg and 3 mL/kg controlled-release nifedipine solution(0. 3 g/L) were poured into stomach to rats in control group, normal dose group and low dose group once everyday for 15 days respectively. Eyeball was removed to collect blood and heart and lung were removed at the same time to test the content of NO and iNOS in serum after last administration.MAIN OUTCOME MEASURES: Content of NO, comparison of iNOS activity between groups.RESULTS: There was significant difference on NO content between normal dose group and control group, low dose group after 15 days' administration ( P < 0. 01 ) . Compared with control group and low dose group, there was significant difference on the iNOS activity in heart and lung of normal dose group(P <0.01, P <0.05).CONCLUSION: While nifedipine reduces the blood pressure, it can also improve the content of NO and resist the increasing activity of iNOS casued by increased blood pressure.

Objective To investigate the changes of TLR4 and nuclear factor kappa B(NF-κB) levels in peripheral blood mononuclear cells(PBMC) of the patients with ulcerative colitis(UC) and its possible pathogenesis mechanism.Methods In our hospital from January 2014 to January 2015,30 cases of UC were selected as the observation group,and other 10 individuals undergoing healthy physical examination in this hospital were selected as the control group.Flow cytometry was used to detect the positive expression rate of TLR4 on surface of peripheral blood CD14+ mononuclear cells.The levels of TLR4,MyD88,NF-κB(P65)mRNA expression were detected by RT-PCR.Western blot was adopted to detect the levels of TLR4,MyD88,NF-κB(P65) protein.Results The positive expression rate of TLR4 on the surface of peripheral blood CD14+ mononuclear cells in the observation group was significantly higher than that in the control group(P＜0.05);the levels of TLR4,MyD88 and NF-κB(P65) B mRNA and protein in the control group were significantly lower than those in the observation group(P＜0.05);the levels of TLR4,MyD88 and NF-κB(P65) protein were positively correlated with the severity of UC.Conclusion The levels of TLR4 and NF-κB in PBMC of the patients with UC are significantly increased,clinic can judge the UC development degree by detecting TLR4 and NF-κB levels.

Objective To study the anti-injury effects of grape seed proanthocyanidin (GSP) on the umbilical vein endothelial cell (HUVEC)injured by hydrogen peroxide(H2O2) and its mechanism. Method Oxidative damage model of endothelial cell was induced by H2O2,and the cells were divided into five groups,including normal groups,oxidative damage group and three (100,50,10 ?g/ml) GSP pretreated groups. MTT assay was used to detect the protection of GSP,Reverse transcription PCR (RT-PCR) was used to detect the protein expression in mRNA level of monocyte chemoattractant protein (MCP-1) and flow cytometry(FCM) was used to detect apoptosis rate and quantitate the expression of intercellular adhesion molecule(ICAM-1)and vascular cell adhesion molecule ( VCAM-1). Results The cell optical density in oxidation damaged group was lower than that in normal control. When pretreated by GSP,the OD value was increased,and was insignificantly different between high GSP group and normal control. In H2O2 damage group,the expression of MCP-1 mRNA and the apoptosis rate were increased,and the numbers of positive cells which express ICAM-1 or VCAM-1 were obviously more than those in pretreated groups respectively.Futhermore,these effects were dose dependent. Conclusion GSP has the protective effect on oxidative damage of HUVEC. The mechanism may be related to stabilization of cell wall and inhibition of inflammatory reaction and apoptosis induced by ICAM-1,VCAM-1 and MCP-1.

Adhesion molecules(AMs) is one kind of glycoproteins expressed on cell surface that can induce interactions between cells or cell and ECM. AMs are indicated to be involved in many chronic complications of diabetes mellitus. Thus, drugs that can decrease levels of AMs are potential new prevention and therapy of diabetic angiopathy.-

Objective 4-Hydroxyisoleucine (4-HIL) is the active component isolated from the seeds of Trigonella foenum-graecum. It exerts broad pharmacological effects such as anti-hyperglycemia,blood lipid lowing,improvement of insulin resistance,etc. As the Chinese traditional medicine,it can be used clinically in treating many diseases such as type 2 diabetes mellitus,dyslipidemia,metabolic syndrome,fatty liver and other diseases. Here the current research status and application prospect on 4-Hydroxyisoleucine were summarized.

Objective To investigate the cell cycle of airway smooth muscle cells(ASMC) in asthmatic rats with airway remodeling and to explore a new method for the prevention and treatment of asthma.Methods The asthmatic rat models were established.The pathologic changes of inner airway wall and ASMC were determined by observing and analyzing the lung tissue sections stained with HE,and the cell cycle distribution of ASMC were analyzed by flow cytometry.Results The inner airway wall and smooth muscle layer were much thicker in rats of asthma group than those of control group(P

Aim To make ethanol from bistortae(EFB)from traditional Chinese medicine bistortae,and investigate the protection on the injury of the human umbiliar vein endothelial cell(ECV304)induced by hydrogen peroxide(H2O2).Methods ECV304 cells were cultured and divided by five experimental groups including normal,injury model induced by H2O2 and three pretreated groups with EFB;flow cytometry(FCM)methods Annexin V/PI staining and PI staining were used to analyze the effect of the medicine on the apoptosis and cell cycle;reverse transcription PCR(RT-PCR)was used to detect the mRNA expression level of apoptosis associated protein caspase-3 in all experiment-al groups.Results FCM Annexin V/PI staining result showed early and advanced stage apoptosis rate was lower when predisposed than that of the damaged group,FCM PI staining showed there were more cells in G0/G1 stage in oxidation damage group,but less in S stage,and hypodiploid peak could be observed,RT-PCR showed that the expression of caspase-3 was weakened when predisposed by EFB than without this disposal(P

BACKGROUND: It has been proved that platelet activation is involved in the development of diabetic angiopathy. Glycoprotein (GP) Ib/IX/V complex is one of the main platelet membrane glycoproteins, and the receptor of both von Willebrand Factor and thrombin. It plays a key role in the process of platelet activation.OBJECTIVE: To observe the expression changes of GP Ib/IX/V complex and its component GP Ibα in patients with type 2 diabetes mellitus. DESIGN: Case-control study. SETTING: Department of Integrated Chinese and Western Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology.PARTICIPANTS: A total of 51 type 2 diabetic outpatients who visited Union Hospital were enrolled from December 2005 to January 2007. The diagnosis was based on the independent criteria from WHO in 1999. Of all the 51 patients, there were 23 females and 28 males, with a peripheral platelet count of over 50×109 L-1. All the subjects had no history of administrating drugs two weeks before the examination, which would potentially influence platelet count. According to disease controlling condition, the patients were assigned to well controlled patient (WCP) group (n = 25) and poorly controlled patient (PCP) group (n =26); and according to whether angiopathy was accompanied, diabetic patients were divided into vascular disease (VD) group (n =27) and non-vascular disease (NVD) group (n =24). Meanwhile 23 healthy subjects were enrolled as normal control group. Informed consents were obtained from subjects and their relatives. The experiments were approved by the ethical committee of Union Hospital.METHODS: Fasting venous blood was harvested from all the subjects' elbow on the early morning of visit day.①Biochemical analysis: Fasting plasma glucose (FPG), glycosylated hemoglobin (HbA1c) and fasting plasma insulin (FINS) was measured by the Clinical Chemistry Department in Union Hospital.② Measurement of platelet membrane GP Ib/IX/V complex and its component GP Ibα: First, 3 mL cubital fasting blood was drawn from each subject and was anti-coagulated with 38 g/L natrium citricum. After that, all samples were fixed with 10 g/L paraform for 45 minutes. Then 50 μL well-fixed blood was added into the polystyrene tube, meanwhile 20 μL monoclonal antibody, such as CD42a-b-c-d and PE-labeled CD42b, was respectively mixed gently with the blood sample and incubated at room temperature in dark for 30 minutes. Next, 20 μL FITC-labeled rat IgG was mixed with the sample containing CD42a-b-c-d and incubated equally. In the end all blood samples were analyzed by FACS420 flow cytometry and the results were expressed as mean fluorescence intensity (MFI). ③ Platelet maximun aggregation rate (MAR) was detected according to reference.MAIN OUTCOME MEASURES: ①Biochemical indicators;②The expressions of GP Ib/IX/V complex and GP Ibα;③Platelet MAR.RESULTS: Fifty-one patients with type 2 diabetes mellitus and twenty-three healthy subjects were all involved in the result analysis.①There were significant differences in FINS in WCP group and PCP group compared with normal controls (P ＜ 0.01). FPG and HbA1c were significantly higher in PCP group compared with normal control group and WCP group (P ＜ 0.01).②Expressions of GP Ib/IX/V complex and GP Ibα were significantly lower in WCP group and PCP group compared with normal control group (P ＜ 0.01), significantly lower in PCP group than in WCP group (P ＜ 0.05), and also significantly lower in VD group and NVD group compared with normal control group (P ＜ 0.01). Moreover the expression of GP Ibα in VD group and NVD group was significantly lower than that of normal control group (P ＜ 0.05), and it also significantly decreased in VD group compared with NVD group (P ＜ 0.05). MFI of GP Ib/IX/V complex had an obvious negative correlation with FBG, HbA1c and FINS (r =-0.634, -0.573, -0.649, P ＜ 0.05), and GP Ibα MFI was obviously negatively correlated with FBG and HbA1c (r =-0.602, -0.543, P ＜ 0.05).③Platelet MAR of diabetic patients were remarkably higher than in healthy subjects (t =-3.852, P ＜ 0.01). Platelet MAR in PCP and VD groups were respectively higher than those in WCP and NVD groups (P ＜ 0.05). CONCLUSION: Platelet activation exists in the early stage of type 2 diabetes mellitus without diabetic angiopathy, and is more obvious after diabetic angiopathy. There is a positive correlation between platelet activation and blood glucose. As a receptor of thrombin and von Willebrand Factor, GP Ib/IX/V complex may be involved in the development of diabetic angiopathy.

AIM: To determine the expression level of Fas molecules on the T cells isolated from systemic lupus erythematosus (SLE) patients. METHODS: The expression of Fas on T cells and the apoptotic rate of 36 patients with SLE in active phase and 18 normal people were determined with flow cytometry. The patients were treated with impact therapy of prednisone, methylprednisolone or cyclophosphamide, respectively, according to their conditions. Within 3 days after admission, immediately before discharging (the average days in hospital is 22.6 d), and 4 weeks after discharging, the amount of Fas molecules expressed on T cell surface was determined respectively with flow cytometry in each patient. RESULTS: There was a positive correlation between SLEDAI, apoptotic rate of T cells and Fas molecules on T cells in SLE patients of active phase (P

Objective To investigate the genotypic and allele frequency differences of melatonin receptor 1B (MTNR1B)-rs4753426 between gestational diabetes mellitus (GDM) pregnancies and normal pregnancies , and to explore the association between single nucleotide polymorphism ( SNP ) of rs4753426 and gestational diabetes mellitus.Methods Totally 93 GDM pregnancies and 165 normal pregnancies were recruited from the Affiliated Hospital of Qingdao University.The age, gestational weeks, height, early pregnant weight , and the levels of fasting plasma glucose ( FPG) , fasting insulin ( FIN) were determined in every participants.By using PCR and DNA sequencing , we detected the distribution of the rs 4753426 genotypes and alleles in all individuals.The homeostasis model assessment-insulin resistance ( HOMA-IR) and the homeostasis model assessment-βcell function ( HOMA-β) were calculated.The allele and genotype frequencies and the FPG , FIN, body mass index ( BMI) , HOMA-IR, HOMA-βlevels between GDM group and control group were compared.Results (1) The genotype frequencies in the GDM group and the control group of rs4753426-CC, CT, TT were 72.0% (67/93), 21.5% (20/93), 6.5% (6/93), and 53.9%(89/165), 40.0% (66/165), 6.1% (10/165) respectively.The allele frequencies in the GDM group and the control group of T and C were 17.2% ( 32/186 ) , 82.8% ( 154/186 ) and 26.1% ( 86/330 ) , 73.9% ( 244/330 ) respectively.There were statistical differences in genotype frequencies and allele frequencies between two groups ( all P<0.05 ).( 2 ) The levels of FPG , FIN and HOMA-IR in the GDM group were obviously higher than those in the control group (P<0.05).The level of HOMA-βwas lower in the GDM group than that of the control group (P<0.05).(3)The FPG of CC and CT genotypes was higher than that of TT genotype in the GDM group (P<0.05), while the level of HOMA-βwas lower than that of TT genotype (P<0.05).Conclusions The MTNR1B-rs4753426 SNP is associated with the pathogenesis of GDM, and rs4753426 is the predisposing locus of GDM.The C-allele is the susceptibility allele of GDM.