BACKGROUND Total hip arthroplasty(THA)is an effective treatment for advanced osteonecrosis of the femoral head,which can significantly relieve pain and improve patients'quality of life.Robotic-assisted THA enhance...BACKGROUND Total hip arthroplasty(THA)is an effective treatment for advanced osteonecrosis of the femoral head,which can significantly relieve pain and improve patients'quality of life.Robotic-assisted THA enhances the accuracy and stability of THA surgery and achieves better clinical outcomes than manual THA.CASE SUMMARY We report the clinical outcomes of robotic-assisted THA and manual THA in the same patient with osteonecrosis of the femoral head.A 49-year-old male patient attended our hospital due to more than 3 years of pain in both hip joints.The left hip was treated with robotic-assisted THA.The patient underwent manual THA of the right hip 3 mo after robotic-assisted THA.We obtained postoperative radiograph parameters,Harris hip score and forgotten joint score of the patient 1 year after surgery.CONCLUSION Compared with manual THA,the patient’s left hip felt better 1 year after roboticassisted THA.Robotic-assisted THA resulted in a better Harris hip score and forgotten joint score than manual THA in the same patient with osteonecrosis of the femoral head.展开更多
Objective To evaluate the role of sclerostin in bone loss of postmenopausal Chinese women with type 2 diabetes mellitus.Methods The postmenopausal patients suffering from type 2 diabetes mellitus and age,body mass ind...Objective To evaluate the role of sclerostin in bone loss of postmenopausal Chinese women with type 2 diabetes mellitus.Methods The postmenopausal patients suffering from type 2 diabetes mellitus and age,body mass index,and duration of menopause matched healthy controls were enrolled into this cross-sectional study according to criteria of inclusion and exclusion.The serum sclerostin level and bone mineral density of the anterior-posterior lumbar spine(L1-L4),femoral neck,and total hip were determined by using a quantitative sandwich ELISA kit and dual X-ray absorptiometry,respectively.Meanwhile,the clinical and laboratory indexes of bone mineral metabolism were analyzed.Associations between serum sclerostin level and bone mineral density as well as bone turnover markers were evaluated by linear regression analysis.Results Finally,265 postmenopausal women with type 2 diabetes and 225 non-diabetic women were recruited in the diabetic group and control group,respectively.Serum sclerostin level of the diabetic group was significantly higher than that of the control group(48.2±19.4 vs.37.2±18.6 pmol/L,P<0.001)and was increased with age in both groups(diabetic group,r=0.374,P<0.001;control group,r=0.312,P<0.001).In type 2 diabetes patients,serum sclerostin concentration was positively correlated with hemoglobin A1c level(r=0.237;P=0.021).Biochemical bone turnover markers,intact parathyroid hormone and bone-specific alkaline phosphatase,were negatively associated with serum sclerostin level(r=0.138,P=0.078 and r=0.265,P<0.001).Conversely,the positive correlation between sclerostin and C-terminal cross-linking telopeptide of type I collagen was found in diabetic patients(r=0.354,P<0.001).Serum sclerostin levels of the diabetic group were positively correlated with bone mineral density of the lumbar spine,femoral neck,and total hip(r=0.324,0.367,and 0.416,respectively;all P<0.001).Conclusions Sclerostin might participate in the pathogenesis of bone loss of type 2 diabetes.The high sclerostin level might serve as a marker of increased osteocyte activity in postmenopausal patients with type 2 diabetes mellitus.展开更多
Objective To explore the mechanisms involved in Staphylococcus aureus(S.aureus) invading human monocytic U937 cells.Methods S.aureus were added to U937 cells at multiplicity of infections(MOI) of 20:1 for 0,15,30,60,a...Objective To explore the mechanisms involved in Staphylococcus aureus(S.aureus) invading human monocytic U937 cells.Methods S.aureus were added to U937 cells at multiplicity of infections(MOI) of 20:1 for 0,15,30,60,and 90 minutes,respectively.Cell apoptosis was analyzed with Hoechst 33258 staining and Annexin V-fluorescein isothiocyanate(FITC)/propidium iodide(PI) flow cytometry analysis.Akt and nuclear factor-κB(NF-κB) activities were detected by Western blotting.Results Infection of U937 cells with S.aureus induced rapid cell death in a time-dependent manner,and the cells displayed characteristic features of apoptosis.S.aureus-induced apoptosis was associated with a prominent downregulation of activated(phosphorylated) Akt and NF-κB.The inhibition of phosphorylated Akt by LY294002 led to the inhibition of NF-κB in a dose-dependent manner.Inhibition of Akt with LY294002 caused further increase in apoptosis of U937 cells.Conclusions S.aureus can stimulate the apoptosis of U937 cells.S.aureus induces apoptosis of U937 cells by inhibiting Akt-regulated NF-κB.展开更多
AIM:To study the morphology and ontogeny of dendritic cells of Peyer's patches in rats at different development periods. METHODS:The morphometric and flow cytometric analyses were performed to detect all the param...AIM:To study the morphology and ontogeny of dendritic cells of Peyer's patches in rats at different development periods. METHODS:The morphometric and flow cytometric analyses were performed to detect all the parameters of villous-crypts axis and the number of OX62+DC, OX62+CD4+SIRP+DC,and OX62+CD4-SIRP-DC in the small intestine in different groups of rats.The relationship between the parameters of villous-axis and the number of DC and DC subtype were analyzed. RESULTS:All morphometric parameters changed significantly with the development of pups in the different age groups(F=10.751,12.374,16.527, 5.291,3.486;P=0.000,0.000,0.000,0.001,0.015). Villous height levels were unstable and increased from 115.24μm to 140.43μm as early as 3 wk postpartum. Villous area increased significantly between 5 and 7 wk postpartum,peeked up to 13 817.60μm2 at 7 wk postpartum.Villous height and crypt depth ratios were relatively stable and increased significantly from 2.80±1.01 to 4.54±1.56,9-11 wk postpartum.The expression of OX62+DC increased from 33.30%± 5.80%to 80%±17.30%,3-11 wk postpartum(F=5.536,P=0.0013).OX62+CD4+SIRP+DC subset levels detected in single-cell suspensions of rat total Peyer's patch dendritic cells(PP-DCs)increased significantly from 30.73%±5.16%to 35.50%±4.08%,5-7 wk postpartum and from 34.20%±1.35%to 43.60%± 2.07%9-11 wk postpartum(F=7.216,P=0.005). CONCLUSION:This study confirms the agerelated changes in villous-crypt axis differentiation in the small intestine.Simultaneously,there are also development and maturation in rat PP-DCs phenotypic expression.Furthermore,the morphological changes of intestinal mucosa and the development of immune cells(especially DC)peaked at 9-11 wk postpartum, indicating that the intestinal mucosae reached a relatively mature state at 11 wk postpartum.展开更多
Objective To elucidate GPR40/FFA1 and its downstream signaling pathways in regulating insulin secretion. Methods GPR40/FFA1 expression was detected by immunofluorescence imaging. We employed linoleic acid (LA), a free...Objective To elucidate GPR40/FFA1 and its downstream signaling pathways in regulating insulin secretion. Methods GPR40/FFA1 expression was detected by immunofluorescence imaging. We employed linoleic acid (LA), a free fatty acid that has a high affinity to the rat GPR40, and examined its effect on cytosolic free calcium concentration ([Ca2+]i) in primary rat β-cells by Fluo-3 intensity under confocal microscopy recording. Downregulation of GPR40/FFA1 expression by antisense oligonucleotides was performed in pancreatic β-cells, and insulin secretion was assessed by enzyme-linked immunosorbent assay. Results LA acutely stimulated insulin secretion from primary cultured rat pancreatic islets. LA induced significant increase of [Ca2+]i in the presence of 5.6 mmol/L and 11.1 mmol/L glucose, which was reflected by increased Fluo-3 intensity under confocal microscopy recording. LA-stimulated increase in [Ca2+]i and insulin secretion were blocked by inhibition of GPR40/FFA1 expression in β-cells after GPR40/FFA1-specific antisense treatment. In addition, the inhibition of phospholipase C (PLC) activity by U73122, PLC inhibitor, also markedly inhibited the LA-induced [Ca2+]i increase. Conclusion LA activates GPR40/FFA1 and PLC to stimulate Ca2+ release, resulting in an increase in [Ca2+]i and insulin secretion in rat islet β-cells.展开更多
Objective To investigate the effects of hepatocyte growth factor(HGF)on vascular endothelial cells apoptosis induced by advanced glycation end products(AGEs)and its possible mechanism. Methods Human umbilical vein end...Objective To investigate the effects of hepatocyte growth factor(HGF)on vascular endothelial cells apoptosis induced by advanced glycation end products(AGEs)and its possible mechanism. Methods Human umbilical vein endothelial cells(HUVECs)were cultured in vitro and intervened by different concentrations of AGEs and HGF.The cell inhibitory rates of each group with different culture time(12, 24, 48, and 72 hours)were measured by methyl thiazolyl tetrazolium(MTT)assay. The early stage apoptosis was detected by flow cytometry with Annexin V-FITC/PI double staining, morphology of cell apoptosis was observed by hoechst 33258 fluorescence staining, and the expression of apoptosis-associated genes Bax and Bcl-2 were determined by Western blotting.The activity of caspase-3 was detected by enzyme-linked immunosorbent assay (ELISA).Results Morphological observation indicated that high concentration of AGEs induced characteristic apoptotic changes in HUVECs.Within a certain concentration range, HUVECs apoptosis inducing rates by AGEs were in both dose- and time-dependent manners.HGF significantly inhibited the apoptosis of HUVECs induced by AGEs (P< 0.05).AGEs significantly promoted expression of Bax protein, but not Bcl-2.Whereas HGF significantly promoted the expression of Bcl-2(P<0.01)and decreased the activity of caspase-3(P<0.05)without affecting Bax level.Conclusions AGEs can induce the apoptosis of endothelial cells in vitro.HGF may effectively attenuate AGEs-induced endothelial cells apoptosis through upregulating Bcl-2 gene expression and inhibiting caspase-3 activation.展开更多
Objective To investigate the effects of stimulant for nucleotide-binding oligomerization domain 1 (NOD1) on secretion of proinflammatory chemokine/cytokines and insulin-dependent glucose uptake in human differentiated...Objective To investigate the effects of stimulant for nucleotide-binding oligomerization domain 1 (NOD1) on secretion of proinflammatory chemokine/cytokines and insulin-dependent glucose uptake in human differentiated adipocytes. Methods Adipose tissues were obtained from patients undergoing liposuction. Stromal vascular cells were extracted and differentiated into adipocytes. A specific ligand for NOD1, was administered to human adipocytes in culture. Nuclear factor-κB transcriptional activity and proinflammatory chemokine/cytokines production were determined by reporter plasmid assay and enzyme-linked immunosorbent assay, respectively. Insulin-stimulated glucose uptake was measured by 2-deoxy-D-[ 3 H] glucose uptake assay. Furthermore, chemokine/cytokine secretion and glucose uptake in adipocytes transfected with small interfering RNA (siRNA) targeting NOD1展开更多
Objective To investigate whether the effect of E.coli on U937 cell lines apoptosis is mediated via p38 mitogen-activated protein kinase (MAPK) activation. Methods The U937 cell lines were treated with E.coli at differ...Objective To investigate whether the effect of E.coli on U937 cell lines apoptosis is mediated via p38 mitogen-activated protein kinase (MAPK) activation. Methods The U937 cell lines were treated with E.coli at different time or together with SB203580, an inhibitor for p38. Cell apoptosis was analyzed by flow cytometry. p38 activities were detected by Western blotting. Results E.coli induced apoptosis in cultured U937 cell lines in a time-dependent manner. The phosphorylation of p38 was induced after 10 minutes infection, reached the peak after 20 minutes, and started to decline after 30 minutes. In contrast, the level of total p38 protein was not changed in whole experimental period. Inhibition of p38 with SB203580 significantly inhibited E.coli induced apoptosis in U937 cells. Conclusion The activation of the p38 MAPK in U937 cell lines by E.coli is a major pathway to mediate the apoptosis.展开更多
Objective To investigate the potential role of nucleotide-binding oligomerization domain 1(NOD1),a component of the innate immune system,in mediating lipid-induced insulin resistance in adipocytes.Methods Adipocytes f...Objective To investigate the potential role of nucleotide-binding oligomerization domain 1(NOD1),a component of the innate immune system,in mediating lipid-induced insulin resistance in adipocytes.Methods Adipocytes from Toll-like receptor 4 deficiency mice were used for stimulation experiments.The effect of oleate/palmitate mixture on nuclear factor-κB(NF-κB)activation was analyzed by reporter plasmid assay.The release of proinflammatory chemokine/cytokines production was determined by using real-time PCR.Insulin-stimulated glucose uptake was measured by 2-deoxy-D-[3H]glucose uptake assay.Chemokine/cytokine expression and glucose uptake in adipocytes transfected with small interfering RNA(siRNA)targeting NOD1 upon fatty acids treatment were analyzed.Results Oleate/palmitate mixture activated the NF-κB pathway and induced interleukin-6,tumor necrosis factor-α,and monocyte chemoattractant protein-1 mRNA expressions in adipocytes from mice deficient in Toll-like receptor 4,and these effects were blocked by siRNA targeting NOD1.Furthermore,saturated fatty acids decreased the ability of insulin-stimulated glucose uptake.Importantly,siRNA targeting NOD1 partially reversed saturated fatty acid-induced suppression of insulin-induced glucose uptake.Conclusion NOD1 might play an important role in saturated fatty acid-induced insulin resistance in adipocytes,suggesting a mechanism by which reduced NOD1 activity confers beneficial effects on insulin action.展开更多
Objective To investigate the inhibitory effect of Tripterygium wilfordii polycoride(TWP) towards the pro-inflammatory factors(TNF-α and IL-1β) on the inflammatory reaction in macrophages induced by LPS and its r...Objective To investigate the inhibitory effect of Tripterygium wilfordii polycoride(TWP) towards the pro-inflammatory factors(TNF-α and IL-1β) on the inflammatory reaction in macrophages induced by LPS and its regulatory effect and influence on the inflammation via TLR4/NF-k B. Methods The MTT method was adopted to test the effect of drugs, TWP, dexamethasone(DXM) and azathioprine(AZA) on cell growth and to select the appropriate concentration. LPS was used to induce the inflammatory reaction in RAW264.7 cell line of mice. Elisa kit was adopted to test the levels of TNF-α and IL-1β. Western blotting was adopted to test the protein expression of TNF-α and IL-1β. RT-PCR was adopted to test the expression of TLR4 and NF-κB. Results The inhibiting effect of TWP on the release of TNF-α and IL-1β in a dose dependent manner. The inhibitory effect of three different TWP dose groups is weaker than that in DXM group. However, TWP in high dose is better than AZA on TNF-α and is as strong as AZA on IL-1β. The dose dependent manner also exits in the effect on the expression of TLR4 and NF-κB, the effect is not weaker, but even stronger than that of DXM and AZA. Conclusion The research shows that down regulation of TLR4 and NF-k B p65 may be one of the mechanisms about the TWP inhibitory effect on TNF-α and IL-1β.展开更多
Central vein catheter (CVC) placement, which is widely utilized in clinical departments, is also highly important in preoperative preparations for radiofrequency catheter ablation (RFCA). The internal jugular vein...Central vein catheter (CVC) placement, which is widely utilized in clinical departments, is also highly important in preoperative preparations for radiofrequency catheter ablation (RFCA). The internal jugular vein or subclavian vein is the routine target. Arterial injury is not uncommon during the procedure but is potentially devastating in spite of the safety and advantages of the Seldinger technique. Compressing of the airway by a hematoma, as well as hemothorax, pseudoaneurysm, arteriovenous fistula, stroke, and even death, has been well-described.展开更多
Objective To study the protective effect of Tripterygium wilfordii polycoride(TWP) against TNBS/ethanol-induced ulcerative colitis(UC) rat model. Methods TNBS and ethanol enema were adopted to build TNBS/ethanol-i...Objective To study the protective effect of Tripterygium wilfordii polycoride(TWP) against TNBS/ethanol-induced ulcerative colitis(UC) rat model. Methods TNBS and ethanol enema were adopted to build TNBS/ethanol-induced UC rat model. Ninety male Wistar rats were divided into six groups: normal, model, low-, medium-, high-dose TWP and azathioprine(AZA) groups, each for 15 rats. All rats were administered by corresponding medicine for 14 d. After 14 d, corresponding colon tissues underwent general and microscopic evaluation. Blood samples were taken from heart and serum was separated by centrifugation. MDA, SOD, GSH, IL-1β and TNF-α levels in serum were tested by ELISA. Colonic samples underwent RT-PCR and Western blotting analysis. Results DAI, general and microscopic evaluation all showed that TWP could promote colonic mucosa healing and such effect was equal to AZA. ELISA results about lipid peroxidation indicated that TWP could decrease MDA level and increase SOD and GSH levels in a dose-dependent manner. TWP with high dose could strongly decrease the MDA level and increase the SOD and GSH levels(P 〈 0.01). ELISA results about inflammatory cytokines indicated that TWP could inhibit the expression of IL-1β and TNF-α in a dose-dependent manner. Western blotting analysis and RT-PCR all indicated that no matter in mR NA level or protein level, TWP could inhibit the expression of NF-κB, TNF-α, IL-1β, and IFN-γ in a dose-dependent manner. The inhibitory effect of AZA towards NF-κB was slightly weaker than TWP with high dose(P 〉 0.05), whereas slightly stronger towards terminal inflammatory cytokines(P 〉 0.05). Conclusion TWP could significantly lower the infiltration of inflammatory cells under microscope, eventually led to mucosa healing, the mechanism of which was to inhibit lipid peroxidation, then further inhibit NF-κB activation, eventually lower the expression of inflammatory meditors locally and systemically.展开更多
Radiofrequency catheter ablation (RFCA),albeit an effective therapy for drug-refractory atrial fibrillation (AF),can be associated with complications in 3.9-22.0% of cases, of which cerebrovascular embolization,ca...Radiofrequency catheter ablation (RFCA),albeit an effective therapy for drug-refractory atrial fibrillation (AF),can be associated with complications in 3.9-22.0% of cases, of which cerebrovascular embolization,cardiac tamponade,and pulmonary vein stenosis are more common,and atrial hematoma is more rare.Here,we presented a case describing an elderly woman diagnosed with an intramural left atrial hematoma after AF ablation.展开更多
基金Supported by the Hangzhou Municipal Health Commission,No.20220119。
文摘BACKGROUND Total hip arthroplasty(THA)is an effective treatment for advanced osteonecrosis of the femoral head,which can significantly relieve pain and improve patients'quality of life.Robotic-assisted THA enhances the accuracy and stability of THA surgery and achieves better clinical outcomes than manual THA.CASE SUMMARY We report the clinical outcomes of robotic-assisted THA and manual THA in the same patient with osteonecrosis of the femoral head.A 49-year-old male patient attended our hospital due to more than 3 years of pain in both hip joints.The left hip was treated with robotic-assisted THA.The patient underwent manual THA of the right hip 3 mo after robotic-assisted THA.We obtained postoperative radiograph parameters,Harris hip score and forgotten joint score of the patient 1 year after surgery.CONCLUSION Compared with manual THA,the patient’s left hip felt better 1 year after roboticassisted THA.Robotic-assisted THA resulted in a better Harris hip score and forgotten joint score than manual THA in the same patient with osteonecrosis of the femoral head.
文摘Objective To evaluate the role of sclerostin in bone loss of postmenopausal Chinese women with type 2 diabetes mellitus.Methods The postmenopausal patients suffering from type 2 diabetes mellitus and age,body mass index,and duration of menopause matched healthy controls were enrolled into this cross-sectional study according to criteria of inclusion and exclusion.The serum sclerostin level and bone mineral density of the anterior-posterior lumbar spine(L1-L4),femoral neck,and total hip were determined by using a quantitative sandwich ELISA kit and dual X-ray absorptiometry,respectively.Meanwhile,the clinical and laboratory indexes of bone mineral metabolism were analyzed.Associations between serum sclerostin level and bone mineral density as well as bone turnover markers were evaluated by linear regression analysis.Results Finally,265 postmenopausal women with type 2 diabetes and 225 non-diabetic women were recruited in the diabetic group and control group,respectively.Serum sclerostin level of the diabetic group was significantly higher than that of the control group(48.2±19.4 vs.37.2±18.6 pmol/L,P<0.001)and was increased with age in both groups(diabetic group,r=0.374,P<0.001;control group,r=0.312,P<0.001).In type 2 diabetes patients,serum sclerostin concentration was positively correlated with hemoglobin A1c level(r=0.237;P=0.021).Biochemical bone turnover markers,intact parathyroid hormone and bone-specific alkaline phosphatase,were negatively associated with serum sclerostin level(r=0.138,P=0.078 and r=0.265,P<0.001).Conversely,the positive correlation between sclerostin and C-terminal cross-linking telopeptide of type I collagen was found in diabetic patients(r=0.354,P<0.001).Serum sclerostin levels of the diabetic group were positively correlated with bone mineral density of the lumbar spine,femoral neck,and total hip(r=0.324,0.367,and 0.416,respectively;all P<0.001).Conclusions Sclerostin might participate in the pathogenesis of bone loss of type 2 diabetes.The high sclerostin level might serve as a marker of increased osteocyte activity in postmenopausal patients with type 2 diabetes mellitus.
基金Supported by the Doctor Research Start-up Fund of Liaoning province (20081055)a grant from the Education Department of Liaoning province (2009A737)
文摘Objective To explore the mechanisms involved in Staphylococcus aureus(S.aureus) invading human monocytic U937 cells.Methods S.aureus were added to U937 cells at multiplicity of infections(MOI) of 20:1 for 0,15,30,60,and 90 minutes,respectively.Cell apoptosis was analyzed with Hoechst 33258 staining and Annexin V-fluorescein isothiocyanate(FITC)/propidium iodide(PI) flow cytometry analysis.Akt and nuclear factor-κB(NF-κB) activities were detected by Western blotting.Results Infection of U937 cells with S.aureus induced rapid cell death in a time-dependent manner,and the cells displayed characteristic features of apoptosis.S.aureus-induced apoptosis was associated with a prominent downregulation of activated(phosphorylated) Akt and NF-κB.The inhibition of phosphorylated Akt by LY294002 led to the inhibition of NF-κB in a dose-dependent manner.Inhibition of Akt with LY294002 caused further increase in apoptosis of U937 cells.Conclusions S.aureus can stimulate the apoptosis of U937 cells.S.aureus induces apoptosis of U937 cells by inhibiting Akt-regulated NF-κB.
基金Supported by Grants from the National Natural Science Foundation of China,No.30571979
文摘AIM:To study the morphology and ontogeny of dendritic cells of Peyer's patches in rats at different development periods. METHODS:The morphometric and flow cytometric analyses were performed to detect all the parameters of villous-crypts axis and the number of OX62+DC, OX62+CD4+SIRP+DC,and OX62+CD4-SIRP-DC in the small intestine in different groups of rats.The relationship between the parameters of villous-axis and the number of DC and DC subtype were analyzed. RESULTS:All morphometric parameters changed significantly with the development of pups in the different age groups(F=10.751,12.374,16.527, 5.291,3.486;P=0.000,0.000,0.000,0.001,0.015). Villous height levels were unstable and increased from 115.24μm to 140.43μm as early as 3 wk postpartum. Villous area increased significantly between 5 and 7 wk postpartum,peeked up to 13 817.60μm2 at 7 wk postpartum.Villous height and crypt depth ratios were relatively stable and increased significantly from 2.80±1.01 to 4.54±1.56,9-11 wk postpartum.The expression of OX62+DC increased from 33.30%± 5.80%to 80%±17.30%,3-11 wk postpartum(F=5.536,P=0.0013).OX62+CD4+SIRP+DC subset levels detected in single-cell suspensions of rat total Peyer's patch dendritic cells(PP-DCs)increased significantly from 30.73%±5.16%to 35.50%±4.08%,5-7 wk postpartum and from 34.20%±1.35%to 43.60%± 2.07%9-11 wk postpartum(F=7.216,P=0.005). CONCLUSION:This study confirms the agerelated changes in villous-crypt axis differentiation in the small intestine.Simultaneously,there are also development and maturation in rat PP-DCs phenotypic expression.Furthermore,the morphological changes of intestinal mucosa and the development of immune cells(especially DC)peaked at 9-11 wk postpartum, indicating that the intestinal mucosae reached a relatively mature state at 11 wk postpartum.
基金Supported by Grant from Department of Education of Liaoning Province (2008810)
文摘Objective To elucidate GPR40/FFA1 and its downstream signaling pathways in regulating insulin secretion. Methods GPR40/FFA1 expression was detected by immunofluorescence imaging. We employed linoleic acid (LA), a free fatty acid that has a high affinity to the rat GPR40, and examined its effect on cytosolic free calcium concentration ([Ca2+]i) in primary rat β-cells by Fluo-3 intensity under confocal microscopy recording. Downregulation of GPR40/FFA1 expression by antisense oligonucleotides was performed in pancreatic β-cells, and insulin secretion was assessed by enzyme-linked immunosorbent assay. Results LA acutely stimulated insulin secretion from primary cultured rat pancreatic islets. LA induced significant increase of [Ca2+]i in the presence of 5.6 mmol/L and 11.1 mmol/L glucose, which was reflected by increased Fluo-3 intensity under confocal microscopy recording. LA-stimulated increase in [Ca2+]i and insulin secretion were blocked by inhibition of GPR40/FFA1 expression in β-cells after GPR40/FFA1-specific antisense treatment. In addition, the inhibition of phospholipase C (PLC) activity by U73122, PLC inhibitor, also markedly inhibited the LA-induced [Ca2+]i increase. Conclusion LA activates GPR40/FFA1 and PLC to stimulate Ca2+ release, resulting in an increase in [Ca2+]i and insulin secretion in rat islet β-cells.
文摘Objective To investigate the effects of hepatocyte growth factor(HGF)on vascular endothelial cells apoptosis induced by advanced glycation end products(AGEs)and its possible mechanism. Methods Human umbilical vein endothelial cells(HUVECs)were cultured in vitro and intervened by different concentrations of AGEs and HGF.The cell inhibitory rates of each group with different culture time(12, 24, 48, and 72 hours)were measured by methyl thiazolyl tetrazolium(MTT)assay. The early stage apoptosis was detected by flow cytometry with Annexin V-FITC/PI double staining, morphology of cell apoptosis was observed by hoechst 33258 fluorescence staining, and the expression of apoptosis-associated genes Bax and Bcl-2 were determined by Western blotting.The activity of caspase-3 was detected by enzyme-linked immunosorbent assay (ELISA).Results Morphological observation indicated that high concentration of AGEs induced characteristic apoptotic changes in HUVECs.Within a certain concentration range, HUVECs apoptosis inducing rates by AGEs were in both dose- and time-dependent manners.HGF significantly inhibited the apoptosis of HUVECs induced by AGEs (P< 0.05).AGEs significantly promoted expression of Bax protein, but not Bcl-2.Whereas HGF significantly promoted the expression of Bcl-2(P<0.01)and decreased the activity of caspase-3(P<0.05)without affecting Bax level.Conclusions AGEs can induce the apoptosis of endothelial cells in vitro.HGF may effectively attenuate AGEs-induced endothelial cells apoptosis through upregulating Bcl-2 gene expression and inhibiting caspase-3 activation.
基金Supported by Grant from Department of Education of Liaoning Province(2008810)
文摘Objective To investigate the effects of stimulant for nucleotide-binding oligomerization domain 1 (NOD1) on secretion of proinflammatory chemokine/cytokines and insulin-dependent glucose uptake in human differentiated adipocytes. Methods Adipose tissues were obtained from patients undergoing liposuction. Stromal vascular cells were extracted and differentiated into adipocytes. A specific ligand for NOD1, was administered to human adipocytes in culture. Nuclear factor-κB transcriptional activity and proinflammatory chemokine/cytokines production were determined by reporter plasmid assay and enzyme-linked immunosorbent assay, respectively. Insulin-stimulated glucose uptake was measured by 2-deoxy-D-[ 3 H] glucose uptake assay. Furthermore, chemokine/cytokine secretion and glucose uptake in adipocytes transfected with small interfering RNA (siRNA) targeting NOD1
文摘Objective To investigate whether the effect of E.coli on U937 cell lines apoptosis is mediated via p38 mitogen-activated protein kinase (MAPK) activation. Methods The U937 cell lines were treated with E.coli at different time or together with SB203580, an inhibitor for p38. Cell apoptosis was analyzed by flow cytometry. p38 activities were detected by Western blotting. Results E.coli induced apoptosis in cultured U937 cell lines in a time-dependent manner. The phosphorylation of p38 was induced after 10 minutes infection, reached the peak after 20 minutes, and started to decline after 30 minutes. In contrast, the level of total p38 protein was not changed in whole experimental period. Inhibition of p38 with SB203580 significantly inhibited E.coli induced apoptosis in U937 cells. Conclusion The activation of the p38 MAPK in U937 cell lines by E.coli is a major pathway to mediate the apoptosis.
基金Supported by the Grant from the Educational Department of Liaoning Province(2008810)
文摘Objective To investigate the potential role of nucleotide-binding oligomerization domain 1(NOD1),a component of the innate immune system,in mediating lipid-induced insulin resistance in adipocytes.Methods Adipocytes from Toll-like receptor 4 deficiency mice were used for stimulation experiments.The effect of oleate/palmitate mixture on nuclear factor-κB(NF-κB)activation was analyzed by reporter plasmid assay.The release of proinflammatory chemokine/cytokines production was determined by using real-time PCR.Insulin-stimulated glucose uptake was measured by 2-deoxy-D-[3H]glucose uptake assay.Chemokine/cytokine expression and glucose uptake in adipocytes transfected with small interfering RNA(siRNA)targeting NOD1 upon fatty acids treatment were analyzed.Results Oleate/palmitate mixture activated the NF-κB pathway and induced interleukin-6,tumor necrosis factor-α,and monocyte chemoattractant protein-1 mRNA expressions in adipocytes from mice deficient in Toll-like receptor 4,and these effects were blocked by siRNA targeting NOD1.Furthermore,saturated fatty acids decreased the ability of insulin-stimulated glucose uptake.Importantly,siRNA targeting NOD1 partially reversed saturated fatty acid-induced suppression of insulin-induced glucose uptake.Conclusion NOD1 might play an important role in saturated fatty acid-induced insulin resistance in adipocytes,suggesting a mechanism by which reduced NOD1 activity confers beneficial effects on insulin action.
基金National Natural Science Foundation of China Granted Project(81273903)
文摘Objective To investigate the inhibitory effect of Tripterygium wilfordii polycoride(TWP) towards the pro-inflammatory factors(TNF-α and IL-1β) on the inflammatory reaction in macrophages induced by LPS and its regulatory effect and influence on the inflammation via TLR4/NF-k B. Methods The MTT method was adopted to test the effect of drugs, TWP, dexamethasone(DXM) and azathioprine(AZA) on cell growth and to select the appropriate concentration. LPS was used to induce the inflammatory reaction in RAW264.7 cell line of mice. Elisa kit was adopted to test the levels of TNF-α and IL-1β. Western blotting was adopted to test the protein expression of TNF-α and IL-1β. RT-PCR was adopted to test the expression of TLR4 and NF-κB. Results The inhibiting effect of TWP on the release of TNF-α and IL-1β in a dose dependent manner. The inhibitory effect of three different TWP dose groups is weaker than that in DXM group. However, TWP in high dose is better than AZA on TNF-α and is as strong as AZA on IL-1β. The dose dependent manner also exits in the effect on the expression of TLR4 and NF-κB, the effect is not weaker, but even stronger than that of DXM and AZA. Conclusion The research shows that down regulation of TLR4 and NF-k B p65 may be one of the mechanisms about the TWP inhibitory effect on TNF-α and IL-1β.
文摘Central vein catheter (CVC) placement, which is widely utilized in clinical departments, is also highly important in preoperative preparations for radiofrequency catheter ablation (RFCA). The internal jugular vein or subclavian vein is the routine target. Arterial injury is not uncommon during the procedure but is potentially devastating in spite of the safety and advantages of the Seldinger technique. Compressing of the airway by a hematoma, as well as hemothorax, pseudoaneurysm, arteriovenous fistula, stroke, and even death, has been well-described.
基金National Natural Science Foundation of China Granted Project(No.81273903 and No.81673798)
文摘Objective To study the protective effect of Tripterygium wilfordii polycoride(TWP) against TNBS/ethanol-induced ulcerative colitis(UC) rat model. Methods TNBS and ethanol enema were adopted to build TNBS/ethanol-induced UC rat model. Ninety male Wistar rats were divided into six groups: normal, model, low-, medium-, high-dose TWP and azathioprine(AZA) groups, each for 15 rats. All rats were administered by corresponding medicine for 14 d. After 14 d, corresponding colon tissues underwent general and microscopic evaluation. Blood samples were taken from heart and serum was separated by centrifugation. MDA, SOD, GSH, IL-1β and TNF-α levels in serum were tested by ELISA. Colonic samples underwent RT-PCR and Western blotting analysis. Results DAI, general and microscopic evaluation all showed that TWP could promote colonic mucosa healing and such effect was equal to AZA. ELISA results about lipid peroxidation indicated that TWP could decrease MDA level and increase SOD and GSH levels in a dose-dependent manner. TWP with high dose could strongly decrease the MDA level and increase the SOD and GSH levels(P 〈 0.01). ELISA results about inflammatory cytokines indicated that TWP could inhibit the expression of IL-1β and TNF-α in a dose-dependent manner. Western blotting analysis and RT-PCR all indicated that no matter in mR NA level or protein level, TWP could inhibit the expression of NF-κB, TNF-α, IL-1β, and IFN-γ in a dose-dependent manner. The inhibitory effect of AZA towards NF-κB was slightly weaker than TWP with high dose(P 〉 0.05), whereas slightly stronger towards terminal inflammatory cytokines(P 〉 0.05). Conclusion TWP could significantly lower the infiltration of inflammatory cells under microscope, eventually led to mucosa healing, the mechanism of which was to inhibit lipid peroxidation, then further inhibit NF-κB activation, eventually lower the expression of inflammatory meditors locally and systemically.
文摘Radiofrequency catheter ablation (RFCA),albeit an effective therapy for drug-refractory atrial fibrillation (AF),can be associated with complications in 3.9-22.0% of cases, of which cerebrovascular embolization,cardiac tamponade,and pulmonary vein stenosis are more common,and atrial hematoma is more rare.Here,we presented a case describing an elderly woman diagnosed with an intramural left atrial hematoma after AF ablation.