Objective: To investigate whether insulin stimulates the translocation of glucose transporter-4 (GLUT4) and glucose uptak e in ischemic myocardium. Methods: Plasma concentration of gluc ose, lactate, free fatty acid and insulin were determined by autoanalyser, and G LUT4 was studied by Western blotting analysis. Results: Insulin increased GLUT4 significantly in sarcolemma of ischemic myocardium ［(25±4)% vs (40±6)%］, and GLUT4 content in intracellular membrane decreased proporti onally. The glucose uptake increased significantly in insulin-ischemic myocardi um. The uptake of insulin-ischemic myocardium was almost 2 times that of ischem ic myocardium. Conclusion: Insulin stimulation results in GLUT4 translocation and increases glucose uptake in ischemic myocardium. When myocardi al ischemia occurs, insulin is helpful in increasing myocardial glucose uptake a nd utilization.

OBJECTIVETo investigate the relationship between the polymorphism of SG13S114 A/T in the 5-lipoxygenase-activating protein (ALOX5AP) gene and the stability of carotid atherosclerosis.

METHODSPolymorphism of SG13S114 A/T in the ALOX5AP gene was analyzed in 152 cases of acute infarction with stable plaque, and 132 cases of acute infarction with vulnerable plaques, by using polymerase chain reaction and restriction fragment length polymorphism. Carotid artery plaque was analyzed by carotid artery color ultrasound.

RESULTSThe frequencies of SG13S114 AA genotype and the A allele in the vulnerable plaque group were higher than that in the stable plaque group (P< 0.01).

CONCLUSIONThe polymorphism of SG13S114 A/T in the ALOX5AP gene may be associated with the instability of atherosclerosis. And the SG13S114 A allele may be a risk factor of vulnerable plaques.

5-Lipoxygenase-Activating Proteins ; Aged ; Aged, 80 and over ; Carotid Artery Diseases ; genetics ; Carrier Proteins ; genetics ; Case-Control Studies ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Male ; Membrane Proteins ; genetics ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide

Objective To investigate the clinical effect of sitagliptin therapy in patients with type 2 diabetes and its effect on glucose trans-porter-4 ( GLUT4).Methods Sixty four patients with type 2 diabetes were randomly divided into two groups , and patients in the observation group (n=32) were given sitagliptin 100 mg qd orally, and those in the control group ( n=32 ) were given acarbose 50 mg tid within meals.The treatment lasted for 12 weeks.The data of the clinical efficacy of the two groups were compared before and after treatment.While 30 healthy controls were selected , only given health education.Data of the expre-ssion of GLUT4 in serum of three groups were detected.Results In ob-servation group , the data of fasting blood glucose ( FBG) , 2 h postprandi-al blood glucose(2 h PG)and glycosylated hemoglobin(HbA1c) were ob-viously better than those in the control group [(6.32 ±0.44 ),(8.76 ± 0.93),(6.85 ±0.37)mmol· L-1 vs(6.44 ±0.46),(9.15 ±0.94), (6.98 ±0.28)mmol· L-1],(P<0.05).The islet beta cell function index in observation group was improved significantly compared with the control group [(2.59 ±0.22),(66 ±18) vs(2.78 ±0.39),(62 ±13)], (P<0.05).The data of serum levels of GLUT4 in the observation group was significantly higher than that in control group [(6.07 ±0.59)vs(2.58 ±0.45)μg· L-1](P <0.05).Conclusion Sitagliptin can effectively control the level of blood sugar , improve islet beta cell function , and the effect may be related to the up -regulation of GLUT4 ex-pression.

OBJECTIVETo observe the injury on micro-skin induced by a self designed micro-skin machine.

METHODSMicro-skin was produced either with the machine or by hand. Cells at the edge of micro-skin were observed by transmission electron microscope. succinic dehydrogenase activity in supernatant of cultivated cells was analyzed, and the cell proliferation of micro-skin was assessed by (3)H-TdR. Twenty patients were enrolled in the study for the observation of the wound healing time between the two groups of micro-skin after being grafted.

RESULTSTransmission electron microscope examination revealed that the cellular injury at the edge of the micro-skin in machine-made group was mild compared with that in man-made group. (3)H-TdR rate was elevated but the activity of succinic dehydrogenase in the supernatant of cultured cells decreased in supernatant of cultured cells of machine produced micro-skin. Wound healing time was shortened in machine made group. (P < 0.05).

CONCLUSIONThe cellular injury at the edge of micro-skin in the machine made group was mild when compared with that in the man-made group with cell proliferation accelerated and wound healing time shortened.

Burns ; surgery ; Cell Division ; Epithelium ; pathology ; Humans ; Microscopy, Electron ; Skin ; ultrastructure ; Skin Transplantation ; methods ; Wound Healing

OBJECTIVETo study the quantity and function of bone marrow (BM) Th17 cells in the blood cytopenia patients with positive BMMNC-Coombs test (IRP) and to explore the role of Th17 cells in the pathogenesis of the disease.

METHODSForty-three untreated IRP patients, 34 recovered IRP patients and 13 healthy donors were enrolled in this study. The ratio of IL-23R(+)CD4(+)/CD4(+)cells in BM were examined by flow cytometry(FCM), the levels of IL-6, IL-23, IL-17 by ELISA, and the expressions of RORγt mRNA, STAT3 mRNA in BMMNC by semiquantitive RT-PCR.

RESULTSThe ratio of IL-23R(+)CD4(+)/CD4(+)cells [(3.6 ± 3.1)%], the levels of IL-6[(26.21 ± 14.55) µg/L], IL-23[(2.23 ± 0.99) µg/L], IL-17[(2.54 ± 1.33) µg/L] and the expressions of RORγt mRNA (0.25 ± 0.08) and STAT3 mRNA (1.10 ± 0.16) in BMMNC of untreated IRP patients were significantly higher than those of recovered IRP patients[(2.0 ± 1.0)%, (9.08 ± 6.36) µg/L, (0.91 ± 0.76) µg/L, (1.28 ± 0.18) µg/L, 0.12 ± 0.08, 0.97 ± 0.12 respectively] (P < 0.05); there was no significiant difference between those of recovered IRP patients and normal controls [(1.9 ± 1.4)%, (14.63 ± 7.66) µg/L, (1.19 ± 0.98) µg/L, (1.50 ± 0.28) µg/L, 0.07 ± 0.05, 0.95 ± 0.13, respectively] (P > 0.05). In IRP group, there were significantly positive correlations between the ratios of IL-23R(+)CD4(+)/CD4(+)cells and CD5(+)CD19(+)/CD19(+) (P < 0.05), there were significantly positive correlations between the levels of IL-17 and CD5(+)CD19(+)/CD19(+), the quantity of BMMNC-antibody (r = 0.494 and 0.377, respectively) (P < 0.05); and so did between the expressions of RORγt mRNA and the ratio of CD5(+)CD19(+)/CD19(+), the quantity of BMMNC-antibody (r = 0.741 and 0.541, respectively) (P < 0.05), and between the ratio of IL-23R(+)CD4(+)/CD4(+) and the level of IL-17, the expression of STAT3 mRNA (r = 0.438 and 0.448, respectively) (P < 0.05).

CONCLUSIONSThere exists increased qunantity and hyperfunction of Th17 cells in the IRP patients which induce B cells hyperfunction and production of autoantibodies against the BM hematopoietic cells. Th17 cells might be a potential new therapeutic target of IRP.

Coombs Test ; Humans ; Interleukin-17 ; Interleukin-6 ; Pancytopenia ; immunology ; Th17 Cells

OBJECTIVETo investigate the effect of vacuum-assisted closure (V. A. C.) technology in the treatment of infected wound of skin and soft tissue as a result of explosion injury in pig.

METHODSSixteen explosion wounds were established by electric detonators on the shoulders and hips on both sides of 4 small white domestic pigs ,and they were divided into A group [(without treatment and infection occurred on 1-2 post burn day (PBD), then treated with vaseline gauze on 3 (PBD)], and B group (with the same treatment as in A group, except for treatment of vacuum assisted closure (V. A. C) with pressure of - 15 kPa after 3 PBD). The data of wound depth, wound area, wound healing time were collected and analyzed at 3 PAD and 1, 3, 6, 9, 14, 19, 24 days after treatment. Specimens from wounds were collected for histopathology observation, including also cell proliferation index, the number of vascular endothelial cells, the activity of myeloperoxidase (MPO) and the number of bacteria.

RESULTSCompared with those in A group on land 3 days after treatment, wound area, wound depth were not enlarged or deepened in B group, while the number of inflammatory cells, vascular endothelial cells, proliferative cells were increased, the activity of MPO was enhanced and the number of bacteria was decreased. There were obvious differences between two groups in following indices: wound area ,wound depth, the number of vascular endothelial cells and bacteria during 1 to 19 days after treatment (P < 0.01)), the number of cell proliferation from 1 - 9 days after treatment (P < 0.01)), and the activity of MPO on 3, 6 days after treatment (P < 0. 01). The wound healing time was (32.8 +/- 1.6) d in A group, which was longer than that in B group (25.8 +/- 1.0 d, P < 0.01).

CONCLUSIONCompared with conventional dressing change, V. A. C can decrease bacteria load, lessen secondary necrosis, prompt the inflammatory response, accelerate the formation of granulation tissue, shorten wound healing time in infectious wound of porcine skin and soft tissue resulted from explosion injury.

Animals ; Blast Injuries ; microbiology ; therapy ; Blood Cell Count ; Female ; Male ; Negative-Pressure Wound Therapy ; Skin ; microbiology ; pathology ; Soft Tissue Injuries ; microbiology ; therapy ; Swine ; Wound Healing ; Wound Infection ; prevention & control

OBJECTIVETo investigate the influence of vacuum-assisted closure (VAC) technique on the growth of capillaries in the wound of the pig produced by explosion.

METHODSFour small white pigs were inflicted with 16 explosion wounds [(7.3 +/- 1.0) cm2 in area] on both sides of the buttocks, shoulders and hips by detonation of a specific type of explosive, and the wounds were randomly divided into 2 groups, i. e, control (C, with conventional treatment from 2 post-injury day (PID) on and treatment (T, with VAC treatment after debridement from 2 PID on) groups, with 8 wounds in each group. Wound tissues of 2mm x 2mm x 2mm in size were harvested for pathological examination before treatment and on 1 and 3 post-treatment day (PTU). The differentiation of adventitial cells were examined with light microscope, and the pixel value of desmin positive particles and the luminal area of newly formed capillaries were assessed with Image C software.

RESULTSMost of vessels in the wound of both groups were in elliptic shape when observed in longitudinal section. In C group, few newly formed capillaries vessels with lack of pericytes were observed before treatment and on 1, 3 PTD, then the number began to increase on 6 PTD. In T group, the number of newly formed capillaries with pericytes was increased on 1 PTD, and it continued to increase thereafter. The pixel values of desmin positive particles in C group on 1, 3, and 6 PTD were (91 +/- 54), (199 +/- 85), and (1552 +/- 298), respectively, which were obviously higher than those in T group [(2569 +/- 330), (3984 +/- 377), (9611 +/- 960), P < 0.01]. The area of vessel lumen in C group was (59 +/- 36), (250 +/- 70), and (938 +/- 287) microm2, respectively on 1, 3, and 6 PTD, which was also smaller than those in T group [(818 +/- 234), (4518 +/- 1080), and (9058 +/- 1656) microm2, P < 0.01].

CONCLUSIONCompared with conventional therapy, VAC can not only accelerate the formation of new capillaries, but also enhance the differentiation of pericytes and the process of enwrapping them around the vessels, and increase the luminal area of newly formed capillaries.

Animals ; Blast Injuries ; therapy ; Capillaries ; cytology ; growth & development ; Female ; Male ; Negative-Pressure Wound Therapy ; Neovascularization, Physiologic ; Swine ; Wound Healing

AIM:To explore the role of ginsenoside Rg1 in the growth of degenerative human lumbar nucleus pulposus cells(HNPCs).METHODS:Cultured HNPCs were subjected to oxygen-glucose deprivation(OGD)to mimic the micro-environment of degenerative HNPCs.The morphological changes of the cells in control group and OGD group were observed under optical microscope.The cells were treated with ginsenoside Rg 1 at concentrations of 25,50 and 100 μmol/L.The expression of collagen II and aggrecan at mRNA and protein levels was determined by real -time PCR and Western blot analysis.The cell viability was measured by CCK-8 assay.The mRNA level of Ki67 was detected by real-time PCR.The apoptosis was analyzed by flow cytometry.The activity of caspase-3 was measured by a caspase-3 kit.The ex-pression of Wnt/β-catenin pathway-related proteins was determined by Western blot.Furthermore,the expression of Wnt/β-catenin pathway-related proteins,the cell viability and apoptosis,and the expression of extracellular matrix synthesis pro-teins were assessed after the cells were co-treated with LiCl and 100 μmol/L ginsenoside Rg1.RESULTS:Normal HNPCs attached on the cell culture plate faster, and were almost round with rich cytoplasm.However, the cell adherence was slower,and the cells were long fusiform with decreased cytoplasm after OGD treatment,indicating that the model of degen-erative HNPCs was successfully established.Compared with normal HNPCs,the expression of collagen II and aggrecan at mRNA and protein levels was decreased in OGD group(P<0.05),which was then increased after the cells were treated with ginsenoside Rg1 at 25,50 and 100 μmol/L(P<0.05).Compared with normal HNPCs,the cell viability and Ki67 expression were decreased in OGD group(P<0.05), which were increased after treatment with ginsenoside Rg 1(P<0.05).Meanwhile,the apoptotic rate and caspase-3 activity were significantly increased in OGD-treated cells(P<0.05), which were decreased after treatment with ginsenoside Rg 1(P<0.05).In addition,the activation of Wnt/β-catenin path-way was also inhibited by ginsenoside Rg 1 treatment at dose of 100 μmol/L(P<0.05).LiCl,a Wnt/β-catenin pathway agonist,obviously decreased the protective effects of ginenoside Rg 1 on OGD-induced cells(P<0.05),indicating that the Wnt/β-catenin pathway was involved in the protective effects of ginenoside Rg 1 on degenerative HNPCs.CONCLUSION:Ginsenoside Rg1 promotes growth and extracellular matrix synthesis of degenerative HNPCs through inhibiting Wnt /β-cate-nin pathway.This study will provide a new idea for prevention and treatment of degenerative HNPCs.

OBJECTIVETo explore the infarct sites in patients with inferior wall acute myocardial infarction (AMI) concomitant with ST segment elevation in leads V1-V3 and leads V3R-V5R.

METHODSFive patients diagnosed as inferior, right ventricular, and anteroseptal walls AMI at admission were enrolled. Electrocardiographic data and results of isotope 99mTc-methoxyisobutylisonitrile (MIBI) myocardial perfusion imaging and coronary angiography (CAG) were analyzed.

RESULTSElectrocardiogram showed that ST segment significantly elevated in standard leads II, III, aVF, and leads V1-V3, V3R-V5R in all five patients. The magnitude of ST segment elevation was maximal in lead V1 and decreased gradually from lead V1 to V3 and from lead V1 to V3R-V5R. There was isotope 99mTc-MIBI myocardial perfusion imaging defect in inferior and basal inferior-septal walls. CAG showed that right coronary artery was infarct-related artery.

CONCLUSIONSThe diagnostic criteria for basal inferior-septal wall AMI can be formulated as follows: (1) ST segment elevates > or = 2 mm in lead V1 in the clinical setting of inferior wall AMI; (2) the magnitude of ST segment elevation is the tallest in lead V1 and decreases gradually from lead V1 to V3 and from lead V1 to V3R-V5R. With two conditions above, the basal inferior-septal wall AMI should be diagnosed.

Aged ; Coronary Angiography ; Diagnostic Errors ; Female ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; diagnosis ; diagnostic imaging ; physiopathology ; Radionuclide Imaging

OBJECTIVETo investigate the expressions of STAT5 phosphorylation in CD34(+)CD38(-)CD123(+) bone marrow cells of the patients with myelodysplastic syndromes (MDS), and then evaluate the level of activation of STAT5 associated with cell proliferation in MDS clone cells.

METHODSThe bone marrow mononuclear cells (BMMNC) were extracted from 36 MDS patients and 14 normal controls. The mean fluorescence intensities (MFI) of phosphorylated STAT5(P-STAT5) in CD34(+)CD38(-)CD123(+) and CD34(+)CD38(-)CD123(-)cells, with or without the stimulation of 10 U/ml EPO, were examined by flow cytometry (FCM).

RESULTSWithout stimulation, the P-STAT5 MFI in CD34(+)CD38(-)CD123(+) cells of low/high risk MDS patients was 113.71 ± 67.22/173.05 ± 102.78, which was significantly higher than that of CD34(+)CD38(-)CD123(-) cells (58.84 ± 27.51/68.99 ± 50.42, P < 0.01, P < 0.05) and the normal controls CD34(+)CD38(-)CD123(-) cells (63.06 ± 21.06, P < 0.05), there was no significant difference between the CD34(+)CD38(-)CD123(-) cells of MDS patients and the normal control CD34(+)CD38(-)CD123(-) cells; With the EPO stimulation, the P-STAT5 MFI in CD34(+)CD38(-)CD123(+) cells of low/high risk MDS patients was 144.04 ± 58.11/239.45 ± 152.05, which was significantly higher than that of CD34(+)CD38(-)CD123(-) cells (68.41 ± 25, 10/64.21 ± 23.43, P < 0.01) and the normal controls CD34(+)CD38(-)CD123(-) cells (75.21 ± 27.02, P < 0.01), there was no significant difference between the CD34(+)CD38(-)CD123(-) cells of MDS patients and the normal control CD34(+)CD38(-)CD123(-) cells; The P-STAT5 MFI in the CD34(+)CD38(-)CD123(+) cells of low/high risk MDS patients with or without EPO stimulation were 21.80/28.86, which was significantly higher than that of CD34(+)CD38(-)CD123(-) cells (7.42/5.50, P < 0.01, P < 0.05) and the normal controls CD34(+)CD38(-)CD123(-) cells (6.39, P < 0.05), there was no significant difference between the CD34(+)CD38(-)CD123(-) cells of MDS patients and the normal controls CD34(+)CD38(-)CD123(-) cells; There was no significant difference of P-STAT5 MFI with or without EPO stimulation and the increased P-STAT5 MFI between the CD34(+)CD38(-)CD123(+) cells of low and high risk MDS.

CONCLUSIONSTAT5 associated with cell proliferation was activated in CD34(+)CD38(-)CD123(+) bone marrow cells in MDS, which had more significant reactions to EPO than CD34(+)CD38(-)CD123(-) cells, indicating that CD34(+)CD38(-)CD123(+) bone marrow cells might be the real malignant MDS clone cells in MDS.

Adult ; Aged ; Aged, 80 and over ; Antigens, CD34 ; metabolism ; Bone Marrow Cells ; cytology ; metabolism ; Cell Proliferation ; Cells, Cultured ; Female ; Flow Cytometry ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; metabolism ; pathology ; Phosphorylation ; STAT5 Transcription Factor ; metabolism