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Objective To investigate the effect of high glycolipid on mouse cardiac microvascular endothelial cells (MCMECs),clarify the role of Tom70 (translocase of the outer mitochondrial membrane 70,Tom70) in it,and explore the related mechanism.Methods MCMECs cultured with normal glucose medium were divided into normal glucose group (NG,5.5mmol/L),high glucose group (HG,25mmol/L) and HG combined with high fat group (HG+HF,glucose concentration 25mmol/L,500μmol/L,16h).Then,the expression of Tom70 in MCMECs was knocked down by siRNA,and the HG+HF group was further divided into vehicle group,Scramble siRNA group and Tom70-siRNA group.To further confirm the specific mechanism of Tom70 in MCMEC injury induced by high glycolipid,Tom70-siRNA group was subgrouped into N-acetylcysteine (NAC)-free group and NAG-containing group.Accordingly,the apoptosis levels were measured by flow cytometer,the generation of nitric oxide (NO) was detected by ELISA kit,the expressions of Tom70 were determined by immunofluorescence and qRT-PCR,and the levels of reactive oxygen species (ROS) by DHE staining and ELISA kit.Results The apoptosis level increased after exposure to HG,and the generation of NO decreased (P＜0.05),while merging HF could aggravate these injuries (P＜0.05).Moreover,HG inhibited the expressions of Tom70 and promoted the production of ROS in MCMECs (P＜0.05).Compared with HG alone,and combination with HF significantly inhibited the expression of Tom70,and significantly increased the production of ROS (P＜0.05).Most importantly,compared with the vehicle group and Scramble siRNA group,the intracellular ROS content and apoptosis rate increased in the Tom70-siRNA group,while generation of NO was significantly decreased (P＜0.01).In contrast,these damage effects mentioned above were partially reversed by the application of antioxidants NAC (P＜0.05).Conclusions High fat can further aggravate the damage on diabetic MCMECs and Tom70 could exert its effect against cardiac microvascular endothelial injury induced by diabetes via inhibiting oxidative stress.
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Objective To investigate the effects and mechanisms of perilipin-5 (Plin5) on the apoptosis of mouse cardiac microvascular endothelial cells induced by high fat and high glucose.Methods The mouse cardiac microvascular endothelial cells (MCMECs) cultured with high glucose medium were respectively given 0,100,300 and 500μmol/L palmitic acid for 24 hours.In order to explore the effects and mechanisms of Plin5 on MCMECs injuries induced by high fat and high glucose,MCMECs exposed to 300μmol/L palmitic acid for 24 hours were divided into control group,Scra siRNA group and Plin5 siRNA group.The control group was only treated with transfection reagent,the Scra siRNA group was given treatment of transfection reagent and garbled RNA,the Plin5 siRNA group was given treatment of transfection reagent and Plin5 specific siRNA.In order to further confirm the specific mechanism of Plin5 in high fat/glucose inducing MCMECs injury,MCMECs in Plin5 siRNA group were divided into vehicle group and N-acetyl cysteine (NAC) group,and given the same intervention of high fat.The apoptotic rate was detected by flow cytometry,qRT-PCR and Western blotting were respectively used to detect the mRNA and protein expression of Plin5,and the intracellular reactive oxygen species (ROS) level was tested by DHE staining and ELISA kit.Results The apoptotic rate of MCMECs was increased in a fat concentration-dependent manner (P＜0.05).Compared with 0μmol/L palmitic acid group,the intracellular ROS content and the expression of Plin5 increased significantly in 300μmol/L palmitic acid group (P＜0.05).Compared with the control group and the Scra siRNA group,the intracellular ROS content and apoptotic rate increased significantly in Plin5 siRNA group under the action of 300μmol/L palmitic acid (P＜0.05).Compared with the vehicle group,the intracellular ROS content and apoptotic rate decreased remarkably in NAC group (P＜0.05).Conclusion With inhibition of oxidative stress,Plin5 may reduce the apoptosis of MCMECs induced by high fat and high glucose.
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Objective To compare the tracing effects of radionuclide and barium sulfate on lactulose hydrogen breath test (LHBT), and to explore the value of LHBT combined with radionuclide imaging in the diagnosis of small intestinal bacterial overgrowth (SIBO) in patients with irritable bowel syndrome (IBS).Methods From November 2010 to November 2012, 89 patients (47 males, 42 females;mean age (45.7±12.9) years) with IBS and 13 healthy volunteers (9 males, 4 females;mean age (43.3±8.6) years) were enrolled in this prospective study.All the subjects underwent LHBT combined with radionuclide imaging.Recording the time when the increment of H2 value >0.005‰ and the OCTT of the radionuclide.Four healthy volunteers also underwent LHBT combined with barium sulfate 1 week after radionuclide imaging.The location of barium sulfate was recorded when H2 value increment >0.020‰.Patients with SIBO received rifaximin treatment, and the effect was observed.χ2 test, Pearson correlation analysis and Wilcoxon rank sum test were used to analyze the data.Results (1)In LHBT combined with barium sulfate test, barium sulfate was found still stagnating in small intestine by abdominal X-ray when H2 value increment >0.020‰ in 4 healthy volunteers, and barium sulfate didn′t reach the colon in delayed imaging in 1 patient.(2) The rates of SIBO detected by LHBT in IBS patients and healthy volunteers were significantly different (43.8%(39/89) vs 5/13;χ2=0.133, P=0.716), and those detected by LHBT combined with radionuclide imaging were also significantly different (39.3%(35/89) vs 1/13;χ2=4.970, P=0.026).(3)The time of H2 value increased >0.005‰ correlated well with OCTT in 13 healthy volunteers ((73±31) and (50±19) min;r=0.871, P<0.001) and 54 IBS patients without SIBO ((83±34) and (66±28) min;r=0.735, P<0.001), but there was no correlation in 35 IBS patients with SIBO ((36±30) and (75±30) min;r=0.304, P=0.076).(4)A total of 34 SIBO-positive patients received a rifaximin treatment, with a significant improvement in the frequency of abdominal pain and abdominal distension after the treatment according to Rome Ⅲ diagnostic criteria: 5(4, 6) vs 4(3, 5), 4(1, 6) vs 0(0, 4)(z values:-4.842 and-5.388, both P<0.001).Conclusion LHBT alone is not a valid test for SIBO, and LHBT combined with radionuclide imaging is a good candidate for SIBO diagnosis.
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The treatment effect of primary and metastatic hepatic carcinoma after local intervention?al therapy is closely related to the prognosis of patients. Traditional imaging modalities such as CT, MRI and ultrasound can only provide anatomical information in monitoring treatment response. In recent years, PET/CT has been widely used in monitoring treatment response for tumors. Many studies have compared the effi?cacy of PET/CT with that of traditional imaging modalities in monitoring the response of primary and meta?static hepatic carcinoma after interventional therapy. This review summarizes recent progress in this field.
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<p><b>OBJECTIVE</b>To study the responses of different pancreatic cancer cells to stimulations by nerve growth factor (NGF) and explore the role of Trk-A in such responses.</p><p><b>METHODS</b>Five pancreatic cancer cell lines (MIA-PaCa-2, PANC-1, SW-1990, AsPC-1, and BxPC-3) were exposed to different concentrations of NGF (0, 4, 20, 100, and 500 ng/ml). MTT and Matrigel invasion method were used to observe the changes in the cell proliferation and invasion ability. Trk-A expression in these cells was detected by PCR and Western blotting, and the relations of Trk-A expression to the cell proliferative and invasive abilities following NGF treatment were analyzed.</p><p><b>RESULTS</b>NGF at 100 ng/ml most obviously stimulated the cell proliferation, and PANC-1 cells showed the highest while AsPC-1 cells showed the least sensitivity to 100 ng/ml NGF stimulation. Matrigel invasion test showed that NGF enhanced the invasiveness of PANC-1 and MIA-PaCa-2 cells but produced only limited effect on AsPC-1 cells; the effect of NGF was completely inhibited by the Trk-A inhibitor CEP701. The expression levels of Trk-A mRNA and protein were the highest in PANC-1 cells and the lowest in AsPC-1 cells.</p><p><b>CONCLUSION</b>NGF can enhance the proliferation and invasiveness of pancreatic cancer cells, and this effect is possibly mediated by Trk-A protein.</p>
Subject(s)
Humans , Cell Line, Tumor , Cell Movement , Cell Proliferation , Neoplasm Invasiveness , Nerve Growth Factor , Pharmacology , Pancreatic Neoplasms , Metabolism , Pathology , Receptor, trkA , Genetics , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the expression of cyclooxygenase-2 (cox-2) in the testes and epididymides of adult male rats and its significance.</p><p><b>METHODS</b>Immunohistochemical staining was used to detect the expression and localization of cox-2 in the testicle and epididymal tissues of 40 adult male SD rats.</p><p><b>RESULTS</b>Strong cox-2 immunoreactivity was detected in the epididymides and testes of the rats. In the caput epididymides, cox-2 expressed mainly in the epithelial nuclei and partly in the cytoplasm. Cox-2 was also found positive in the testis nuclei and cytoplasm.</p><p><b>CONCLUSION</b>Immunohistochemical staining is a fairly sensitive method for detecting cox-2 expression in the testes and epididymides of adult male rats.</p>
Subject(s)
Animals , Male , Rats , Cyclooxygenase 2 , Epididymis , Immunohistochemistry , Rats, Sprague-Dawley , Sensitivity and Specificity , TestisABSTRACT
Objective To certify whether 99mTc-HL91 can be trapped in ischemic myocardium prominently for clinical imaging. Methods The model of myocardial ischemia in rabbits was prepared and the distribution of 99mTc-HL91 was observed. The distribution of 99mTc-HL91 in the whole body was also observed. Results The myocardium of the left ventricle supplied by LCX was ischemic according to the change of electrical physiology and the results of NBT dying. The radioactivity in the myocardium of left ventricle supplied by LCX was higher than that supplied by LAD. LCX/LAD: 120 min, 2.57; 180 min, 3.45. 99mTc-HL91 was mostly distributed in the liver and kidney, next, the stomach and intestine. It was excreted through urination and defecation. There was little radioactivity in the heart and lungs and it eliminated quickly. 99mTc-HL91 was eliminated quickly from blood, too. Conclusion Ischemic myocardium can trap 99mTc-HL91 more significantly and relatively prominently at 180 minutes of post-injection. 99mTc-HL91 can be used in myocardial hypoxia imaging.