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Objective:To study the effect of Biejiajian Wan on the epithelial-mesenchymal transition （EMT） of rat hepatic oval cells induced by transforming growth factor- β1（TGF-β1）， in order to explore its mechanism in reversing EMT. Method:WB-F344 cells were divided into five groups： blank group， TGF-β1 model group （10 μg·L-1TGF-β1）， low-dose group （10 μg·L-1TGF-β1+0.55 g·kg-1Biejiajian Wan）， medium-dose group （10 μg·L-1TGF-β1+1.1 g·kg-1Biejiajian Wan）， high-dose group （10 μg·L-1TGF-β1+2.2 g·kg-1Biejiajian Wan）. Except blank group， TGF-β1 was used to induce WB-F344 cells in all of the remaining groups to construct an EMT model. After the cells were treated with low， medium and high doses of Biejiajian Wan serum， the changes of migration ability of WB-F344 cells were detected by cell scratching test. The expressions of E-cadherin， N-cadherin and Vimentin were detected by Western blot. Real-time PCR was used to detect the changes in the expression of β-catenin mRNA. The expression of β-catenin was detected by cell immunofluorescence assay. Result:Compared with normal WB-F344 cells， the intercellular space of WB-F344 cells became loose from tight， and the morphology changed from cobblestone to fibroblast after TGF-β1 induced WB-F344 cells for 4 days， and the expression of E-cadherin protein decreased， while the expression of N-cadherin protein increased （P<0.01）， indicating that the EMT model of WB-F344 cells was successfully built. Compared with the blank group， the migration ability of WB-F344 cells in TGF-β1 model group was enhanced （P<0.01）， compared with TGF-β1 model group， Biejiajian Wan could significantly inhibit the migration ability of WB-F344 cells； specifically， the low-dose group had no statistical significance， and the medium and high-dose groups had statistical significance （P<0.05）. Western blot results showed that compared with the blank group， the expression of E-cadherin decreased， whereas those of N-cadherin and Vimentin increased in the TGF-β1 model group （P<0.01）， compared with TGF-β1 model group， E-cadherin protein expression was increased in the low， medium and high-dose groups， while the expressions of N-cadherin and Vimentin was decreased； specifically， the low-dose groups had no statistical significance， and the medium and high dose groups had statistical significance （P<0.05，P<0.01）. Real-time PCR results showed that compared with the blank group， the mRNA expression of β-catenin in the TGF-β1 model group was increased （P<0.05）， whereas compared with TGF-β1 model group， the mRNA expression of β-catenin in the low， medium and high-dose groups of Biejiajian Wan was reduced （P<0.01）. The results of cellular immunofluorescence showed that compared with the blank group， the fluorescence expression of β-catenin in the cell nucleus was enhanced in the TGF-β1 model group； and compared with the TGF-β1 model group， the expression of β -catenin in the cell nucleus of the low， medium and high-dose groups of Biejiajian Wan decreased， and the inhibitory effect of Biejiajian Wan on β-catenin in the cell nucleus was positively correlated with its concentration. Conclusion:Biejiajian Wan may reverse the EMT process that TGF-β1 induced WB-F344 cells， and inhibit the migration of WB-F344 cells by inhibiting Wnt/β-catenin signaling pathway.
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The present study observed the effect of Guanxin Zhitong Capsules(GXZT) on the lipotoxicity of vascular endothelial cells and investigated the mechanism of GXZT in atherosclerosis treatment. The lipotoxicity model in human umbilical vein endothelial cells(HUVECs) was induced by palmitic acid(PA) stimulation. These cells were divided into a normal control group(NC, 15% normal serum), a model group(PA, 0.6 mmol·L~(-1) PA+15% normal serum), a high-dose GXZT group(GXZT-H, 0.6 mmol·L~(-1) PA+15% GXZT-medicated serum), a medium-dose GXZT group(GXZT-M, 0.6 mmol·L~(-1) PA+10% GXZT-medicated serum+5% normal serum) and a low-dose GXZT group(GXZT-L, 0.6 mmol·L~(-1) PA+5% GXZT-medicated serum+10% normal serum). HUVECs were detected for cell viability by cell counting kit-8(CCK-8) assay, apoptosis by flow cytometry, mitochondrial membrane potential(MMP) by JC-1 labeled laser scanning confocal microscopy, and total and phosphorylated proteins of p38, ERK1/2, and JNK1/2 in the mitogen-activated protein kinases(MAPK) signaling pathway by Western blot. The phosphorylated level was calcula-ted. Compared with the NC group, the PA group showed decreased cell viability and MMP(P<0.01, P<0.01), elevated apoptosis(P<0.01), and up-regulated phosphorylated levels of p38, ERK1/2, and JNK1/2(P<0.01, P<0.01, P<0.01). Compared with the PA group, the GXZT-H, GXZT-M, and GXZT-L groups showed increased cell viability and MMP(P<0.01, P<0.01, P<0.01), reduced apoptosis(P<0.01), and down-regulated protein expression and phosphorylated levels of p38, ERK1/2 and JNK1/2 in the MAPK signaling pathway(P<0.01, P<0.01, P<0.01). In conclusion, the results suggest that GXZT functions via blocking MAPK signaling pathway to relieve the damage of HUVECs induced by PA.
Subject(s)
Humans , Apoptosis , Capsules , Human Umbilical Vein Endothelial Cells/metabolism , MAP Kinase Signaling System , Palmitic Acid/toxicity , Signal Transduction , p38 Mitogen-Activated Protein Kinases/metabolismABSTRACT
@#In most mammalian central nervous system diseases, axons are damaged.Due to the limited ability of damaged neurons to promote axonal regeneration, the formation of glial scar and the release of inhibitory nutrients, it is difficult to regenerate axons of damaged neurons. The purpose of this study was to investigate the effect of cerebroprotein hydrolysate for injection (II) (CBL) on neuritogenesis and its underlying mechanism. Immunofluorescence staining was used to detect the axon length of mouse neuroma cells (Neuro-2a) and mouse primary cortical neuronal cells. Western blotting was used to detect the expression of phosphorylated TrkB protein in Neuro-2a cells and mouse primary cortical neuronal cells. The results showed that CBL could increase the axon length of Neuro-2a cells or mouse primary cortical neuronal cells, and that the phosphorylation level of TrkB in neuronal cells was significantly increased when 5 μg/mL CBL was applied to neuronal cells for 1 h. In conclusion, CBL can promote neuritogenesis, and increase the expression of phosphorylated TrkB, which may be related to the activation of TrkB signaling pathway.
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Objective:To study the effect of Biejia Jianwan on expressions of signal molecules and target genes of transforming growth factor-β (TGF-β)/Smad pathway in diethylnitrosamine (DEN)-induced rat hepatocellular carcinoma, and explore the mechanisms of Biejia Jianwan suppressing the invasion of hepatocellular carcinoma. Method:The rats were divided into three group, namely normal group, model group and Biejia Jianwan group (2.2 g·kg-1·d-1). Rats in Biejia Jianwan group and model group received intraperitoneal injections of DEN to induce sequential chronic inflammation, cirrhosis and hepatocellular carcinoma. At the sign of cirrhosis, rats in Biejia Jianwan group began taking Biejia Jianwan by gavage for 6 weeks. Rat blood was collected to measure serum levels of biochemical markers of liver function tests, including alanine aminotransferase(ALT), aspartate aminotransferase(AST), total bilirubin(TBIL), albumin(Alb), γ-glutamyl transpeptadase(GGT), alkaline phosphatase(ALP). Rat livers were fixed in formalin and stained with hematoxylin-eosin (HE)staining， quantitative real-time PCR was used to test the mRNA expressions of TGF-β1， and Western blot was used to test protein expressions of TGF-β1, Smad2/3, p-Smad2/3, N-cadherin, E-cadherin and Vimentin. Result:All of the levels of biochemical markers showed no difference in Biejia Jianwan group and model group. Biejia Jianwan could improve the pathological changes of balloon-like degeneration, edema, and necrosis in liver cancer tissues. Importantly, the treatment dramatically decreased the mRNA expression of TGF-β1(P<0.01), and the protein expressions of TGF-β1, p-Smad2(P<0.01). Besides, the protein expression of N-cadherin and Vimentin were decreased significantly (P<0.01). Conclusion:Biejia Jianwan can inhibit epithelial-mesenchymal transition (EMT) in hepatocellular carcinoma cells activated via TGF-β/Smad pathway by reducing TGF-β1 expression, so as to suppress the metastasis and invasion of hepatocellular carcinoma.
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Objective@#To understand demand for emergency education among college students and to analyze the influencing factors, to provide the evidence for college to make prevention and controlling measures.@*Methods@#A survey was conducted among college students who were selected by stratified random sampling from 4 colleges in Liaoning Province, and data were analyzed using general descriptive analysis, chi-square test and multivariate Logistic regression analysis.@*Results@#There were 90.7% of college students who had urgent needs for emergency education. Students who were female(93.9%), seniors(94.4%), learning in medical colleges(97.6%), having mothers with higher education levels(92.6%), and living in urban areas(94.4%) had higher educational needs. Multivariate analysis showed that gender (OR=5.00), school category (OR=3.87), emergency attitude (OR=8.02), active learning behavior (OR=3.91), emergency knowledge self-assessment (OR=6.64) were influencing factors(P<0.05).@*Conclusion@#The emergency knowledge and preparation of college students were insufficient and emergency education was needed. The government and schools should strengthen their attention and input, develop more effective ways to disseminate emergency knowledge among students so as to improve their response ability.
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Aim To screen the differential microRNA (miRNA) expression profiles of different metastatic po-tential liver cancer cell lines,and predict miRNAs-reg-ulated target genes and their functions. Methods To-tal RNA was extracted and the miRNA expression pro-files were obtained by miRNA microarray chip hybrid-ization. The miRNAs whose expression had significant difference were selected by analyzing the miRNA difference expression profiles of the two different meta-static potential liver cancer cell lines, namely MHCC-97H(high-metastasis) and Hep3B(non-metastasis), which were compared with normal hepatocytes L02 re-spectively. Moreover, we analyzed the miRNA differ-ential expression profile between liver cancer cell lines MHCC-97H and Hep3B. The miRNAs were verified by qPCR and target genes were predicted by four softwares (TargetScan, miRanda, miRWalk, miRDB). To un-derstand the biological functions of predicted target genes, bioinformatics analysis was performed. Results The miRNA microarray results showed that the ex-pression of miR-192-5p and miR-215-5p significantly increased in liver cancer cell lines (MHCC-97H, Hep3B) when compared with normal hepatocytes L02, while miR-130a-3p and miR-196a-5p were significantly reduced; compared with Hep3B, the expression of miR-224-5p markedly increased in liver cancer cell line MHCC-97H, while miR-146a-5p, miR-483-3p and miR-200b-3p were significantly reduced. The re-sults of qRT-PCR were consistent with chip results. Conclusion There are differences of miRNA expres-sion profiles in different metastatic potential liver canc-er cell lines MHCC-97H, Hep3B, and they may par-ticipate in regulating the development and invasion of hepatocellular carcinoma.
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Invasion and metastasis are key factors contributing to the high mortality rate of patients with hepatocellular carcinoma (HCC) involving a complex mechanism. In the invasion and metastasis of HCC, miRNAs can serve as either oncogenes or tumor suppressor genes to regulate the differentiation and proliferation of tumor cells being and play important roles in tumorigenesis, angiogenesis, invasion and metastasis. This review summarizes the recent progress in research of the molecular mechanisms by which miRNAs targeting GSK-3β regulate HCC invasion and metastasis and examines the roles of miRNAs in hepatocellular carcinoma cell proliferation, apoptosis, invasion, metastasis, and GSK-3β regulation.
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Dimethyl sulfide (DMS) has been historically recognized as a metabolite of the marine microorganism or a disgusting component for the smell of halitosis patients. In our recent study, DMS has been identified as a cytoprotectant that protects against oxidative-stress induced cell death and aging. We found that at near- physiological concentrations, DMS reduced reactive oxygen species (ROS) in cultured PC12 cells and alleviated oxidative stress. The radical-scavenging capacity of DMS at near-physiological concentration was equivalent to endogenous methionine(Met)-centered antioxidant defense. Methionine sulfoxidereductase A (MsrA), the key antioxidant enzyme in Met-centered defense, bound to DMS and promoted its antioxidant capacity via facilitating the reaction of DMS with ROS through a sulfonium intermediate at residues Cys72, Tyr103, Glu115, followed by the release of dimethyl sulfoxide (DMSO). MTT assay and trypan blue test indicated that supplement of DMS exhibited cytopro?tection against 6-hydroxydopamine and MPP + induced cell apoptosis. Furthermore, MsrA knockdown abolished the cytoprotective effect of DMS at near- physiological concentrations. The present study reveals new insight into the potential therapeutic value of DMS in Parkinson disease.
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<p><b>OBJECTIVE</b>To observe the clinical efficacy of Jiakangling Capsule (JC) combined with reduction of 1311 in treatment of Graves hyperthyroidism.</p><p><b>METHODS</b>Totally 387 Graves hyperthyroidism patients were randomly assigned to the treatment group (200 cases) and the control group (187 cases). Patients in the treatment group took JC combined with reduction of 131I. The 131I dosage per gram of thyroid tissue was 50-80 microCi. They additionally took JC one week after taking 1311 for one consecutive month. Patients in the control group took 131 routinely as one disposable treatment. The 131I dosage per gram of thyroid tissue was 70-120 microCi, without using JC or other anti-thyroid drugs. All patients were reexamined after 24-month treatment. Whether hyperthyroidism was cured, incurred, or permanent was observed. Efficacies of thyroglobulin antibody (TGAb) and thyroid microsome antibody (TMAb) were compared between the two groups.</p><p><b>RESULTS</b>Compared with the control group, the incurred ratio increased in the treatment group [3.2% (6/187) vs. 16.0% (32/200), P < 0.01], the incurred ratio of strong positive TGAb and TMAb patients increased [3.5% (2/57) vs. 27.1% (16/59), P < 0.01], the permanent hypothyroidism ratio decreased [21.1% (12/57) vs. 3.4% (2/59), P < 0.05 ].</p><p><b>CONCLUSION</b>JC combined with reduction of 1311 was superior in treating Graves hyperthyroidism induced permanent hypothyroidism than routine 1311 treatment, especially for strong positive TGAb and TMAb patients.</p>
Subject(s)
Humans , Autoantibodies , Capsules , Drugs, Chinese Herbal , Therapeutic Uses , Graves Disease , Drug Therapy , Hyperthyroidism , Drug Therapy , Hypothyroidism , Iodine RadioisotopesABSTRACT
Objective To study the regulatory effects of long non-coding RNA HIF1A-AS1 on the myocardial ischemia reperfusion (I/R) injury and the related mechanism. Methods Myocardial I/R injury model was established with SD rats, and hypoxia reoxygenation (H/R) model was established with rat cardiac myocytes. si-HIF1A-AS1 was used to inhibit HIF1A-AS1 expression in the cardiac myoctyes. Then the mRNA expression of HIF1A-AS1 was detected by real-time PCR, the growth vitality of cardiac myocytes was investigated by MTT assay, the concentration of lactate dehydrogenase (LDH) in the culture media was detected by ELISA, and the autophagy-associated protein Beclin-1 expression was observed by Western blotting analysis. Results HIF1A-AS1 expression was increased in cardiac muscle of rat I/R model and rat cardiac myocytes of H/R model. Inhibition of HIF1A-AS1 by siRNA protected the cardiomyocytes against H/R injuries, reversing the decreased growth vitality of cardiac myoctyes, increased LDH level in the culture media, and increased expression of autophagy-related protein Beclin-1 induced by H/R stimulation. Conclusion Inhibition of long non coding RNA HIF1A AS1 might play a protective role in I/R injury of cardiac myoctyes by inhibiting the excessive autophagy of cardiomyocytes.
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Objective To study the regulatory effect of urocortin (UCN) on ischemia/reperfusion (I/R)-induced myocardial autophagy, so as to explore the myocardial protection mechanism of UCN. Methods Cardiac I/R model was established with rats and hypoxia/reoxygenation(H/R) model was also established with neonatal rat cardiomyocytes. The injury was created by ischemic/hypoxia for 1 h plus reperfusion/reoxygenation for 2 h, and UCN pretreatment was given 1 h before ischemia/hypoxia. The I/R or H/R-induced myocardial injury, myocardial autophagy and autophagy-related gene expression were observed 2 h after reperfusion/reoxygenation. Results UCN pretreatment greatly reduced I/R-induced myocardial damage by decreasing the infarct size, serum creatine kinase (CK) and lactate dehydrogenase (LDH) concentration, increasing the vitality of H/R cardiomyocytes in vitro, and reducing LDH level in the culture supernatant. Moreover, UCN pretreatment also inhibited H/R-induced myocardial autophagy by reducing the ratio of LC3BII/LC3B I and inhibiting expression of autophagy-related genes (Beclin1 and Bnip3). Conclusion UCN can inhibit I/R-induced myocardial autophagy, which may play an important role in the protection against I/R injury.
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The expression profile in the mouse hepatitis B virus X (HBx)-transfected model was investigated in order to lay a foundation for further study on the implication of cytokines expression in hepatitis B virus (HBV) infection. Hydrodynamic injection method via the tail vein was used to establish the animal HBx-transfected model. By using microassay, the differential expression of gene in each group was analyzed, which was further confirmed by using real-time PCR and semi-quantitative PCR. Most of chemokine genes such as Ccl2, Ccl5, Ccl9, MIG and IP-10 were up-regulated in the HBx-transfected mouse model versus the control mice, which was coincided with the microarray results. Western blotting and immunohistochemistry were applied to detect the expression of MIG and IP-10 in the liver tissues. Simultaneously, ELISA was adopted to measure the content of IFN-γ in the liver tissues. DNA microassay revealed that the expression of 611 genes changed in HBx-transfected mice as compared with that in pCMV-tag2B-transfected mice, and most of the screened chemokines were up-regulated (including MIG and IP-10). Additionally, IFN-γ protein levels were increased by 20.7% (P<0.05) in pCMV-tag2B-HBx-transfected mice as compared with the untreated mice. IFN-γ protein levels were reduced by 53.9% (P<0.05) in pCMV-tag2B-transfected mice as compared with the untreated mice, which was consistent with the up-regulation of MIG and IP-10. It was suggested HBx transfection could induce the expression of MIG and IP-10 in the liver tissues, which might play the roles in HBV-related liver immunity and cytokines-mediated antiviral effect.
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The expression profile in the mouse hepatitis B virus X (HBx)-transfected model was investigated in order to lay a foundation for further study on the implication of cytokines expression in hepatitis B virus (HBV) infection. Hydrodynamic injection method via the tail vein was used to establish the animal HBx-transfected model. By using microassay, the differential expression of gene in each group was analyzed, which was further confirmed by using real-time PCR and semi-quantitative PCR. Most of chemokine genes such as Ccl2, Ccl5, Ccl9, MIG and IP-10 were up-regulated in the HBx-transfected mouse model versus the control mice, which was coincided with the microarray results. Western blotting and immunohistochemistry were applied to detect the expression of MIG and IP-10 in the liver tissues. Simultaneously, ELISA was adopted to measure the content of IFN-γ in the liver tissues. DNA microassay revealed that the expression of 611 genes changed in HBx-transfected mice as compared with that in pCMV-tag2B-transfected mice, and most of the screened chemokines were up-regulated (including MIG and IP-10). Additionally, IFN-γ protein levels were increased by 20.7% (P<0.05) in pCMV-tag2B-HBx-transfected mice as compared with the untreated mice. IFN-γ protein levels were reduced by 53.9% (P<0.05) in pCMV-tag2B-transfected mice as compared with the untreated mice, which was consistent with the up-regulation of MIG and IP-10. It was suggested HBx transfection could induce the expression of MIG and IP-10 in the liver tissues, which might play the roles in HBV-related liver immunity and cytokines-mediated antiviral effect.
Subject(s)
Animals , Male , Mice , Chemokine CXCL10 , Allergy and Immunology , Chemokine CXCL9 , Allergy and Immunology , Cytokines , Allergy and Immunology , DNA, Viral , Genetics , Hepatitis B , Genetics , Allergy and Immunology , Virology , Hepatitis B virus , Genetics , Allergy and Immunology , Mice, Inbred C57BL , Mice, Transgenic , Trans-Activators , Genetics , Transfection , MethodsABSTRACT
<p><b>OBJECTIVES</b>To explore the feature of the edge-to-edge technique and its effect for mitral regurgitation due to myxomatous degeneration.</p><p><b>METHODS</b>The in-patient data and follow-up outcomes of 58 patients after the edge-to-edge technique for mitral regurgitation due to myxomatous degeneration from January 2000 to January 2009 were analyzed retrospectively. Of the 58 patients, 32 patients were male and 26 patients were female, and the age range was from 43 years to 65 years with a mean of (56 ± 6) years, and moderate mitral regurgitation was observed in 18 patients and severe regurgitation in 40 patients, and the prolapse of the anterior leaflet was observed in 50 patients and the prolapse of the bileaflet in 8 patients. The edge-to-edge technique was performed in all patients and the annuloplasty was performed in 44 patients.</p><p><b>RESULTS</b>There was no perioperative death and serious complication. Postoperative transthoracic echocardiography of all the survivors indicated that the dimensions of left atrial and left ventricular were obviously decreased (P < 0.05) and mitral insufficiency was obviously improved (no regurgitation was observed in 9 patients and trace regurgitation in 30 patients and mild regurgitation in 19 patients) and there was no mitral stenosis. Totally 58 patients were followed up from 24 months to 95 months with a mean of (58 ± 20) months. During the follow-up, there were 2 deaths for noncardiac factors. Freedom from recurrent moderate or severe mitral regurgitation at 5 years after operations was 91.9%. According to undergoing combined annuloplasty or not, 58 patients were divided into the edge-to-edge technique group (14 cases) and the edge-to-edge technique + annuloplasty group (44 cases), and the survival analysis shows there was significant difference on freedom from long-term recurrent moderate or severe mitral regurgitation after operations between two groups (χ(2) = 4.034, P = 0.045) and long-term effect of the latter group was better.</p><p><b>CONCLUSIONS</b>The edge-to-edge technique can be conveniently used and bring about satisfactory perioperative and long-term effects for mitral regurgitation due to myxomatous degeneration. The combination of the edge-to-edge technique and the annuloplasty can improve the long-term effect significantly.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Follow-Up Studies , Mitral Valve , General Surgery , Mitral Valve Insufficiency , General Surgery , Mitral Valve Prolapse , General Surgery , Retrospective Studies , Suture Techniques , Treatment OutcomeABSTRACT
Objective: To evaluate the safety and efficacy of radiofrequecy ablation for atrial fibrillation during minimally invasive mitral valve surgery via right thoracotomy. Methods: From Jan. 2008 to Dec. 2011, 30 patients underwent radiofrequecy Maze EI procedure for atrial fibrillation during mini-invasive mitral valve surgery (study group). Another 30 patients with atrial fibrillation undergoing mitral valve surgery through median sternotomy without Maze procedure during the same period were taken as controls. The pre-treatment data of the patients were matchable between the two groups. The study group received mitral valve repair/replacement and radiofrequecy Maze HI procedure for atrial fibrillation. The operative outcome, postoperative complication and elimination rate of atrial fibrillation were compared between the two groups. Results: No patient in the study group was transferred to median sternotomy during operation, and there was no reoperation, prolonged incubation, failure of important organs, hemoglobinuria or death. Compared with the control group, the study group had significantly longer mean circulation arrest time and cardiopulmonary bypass time, significantly reduced chest drainage and blood transfusion volume, and significantly shortened hospital study (P<0. 05). The elimination rates of atrial fibrillation at immediately after operation,discharge and 6 months after operation were 96. 7%, 66. 7% and 73. 3% in the study group, and 50%, 23. 3% and 16. 7% in the control group, respectively, with significant difference found between the two groups (P< 0.01). Compared with the control group, better heart function recovery was achieved in the study group at 6 month after operation. Conclusion: Radiofrequecy ablation for atrial fibrillation during minimally invasive mitral valve surgery via right thoracotomy is safe and effective. Importantly, it does not increase risks and complications of surgery. The early and middle term effects are satisfactory.
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Objective: To investigate the inhibitory effects of RNA silencing via adenovirus-mediated vascular endothelial growth factor receptor (VEGFR) shRNA on proliferation of lung adenocarcinoma cells in vitro and in vivo. Methods: Ad-VEGFRshRNA adenovirus containing enhanced green fluorescent protein (EGFP) gene and VEGFRshRNA was constructed and was used to infect A549 cells; fluorescent microscopy was used to observe the infection efficiency. Western blotting assay was used to examine the expression of VEGFR protein in A549 cells. MTT method was used to examine the cell viability and the cell growth curve was drawn. The inhibition of cell growth was examined by cell cycle and colony-forming test. Meanwhile, nude mice were transplanted with A549 cells to establish tumor-bearing model, and the long term growth of tumor was observed. Results: Western blotting revealed that the expression of VEGFR was obviously decreased in the RNA interference group. The cell growth curve indicated that the cell growth was obviously inhibited after RNA interference. Cell cycle and colony-forming test indicated that the tumor growth was obviously inhibited after RNA interference. In vivo study with nude mice also indicated that RNA interference obviously inhibited tumor growth. Conclusion: The constructed VEGFR-targeted shRNA can effectively inhibit VEGFR expression in A549 cells and can suppress the growth of A549 cells.
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<p><b>BACKGROUND</b>Most patients with acute myelogenous leukemia (AML) suffer from disordered hemostasis. We have previously shown that annexin II (Ann II), a high-affinity co-receptor for plasminogen/tissue plasminogen activator, plays a central role in primary hyperfibrinolysis in patients with acute promyelocytic leukemia (APL). The expression of Ann II in cells from patients with major subtypes of AML and the effect of arsenic trioxide (As2O3) on Ann II expression in AML cells were investigated to determine whether As2O3-mediated downregulation of Ann II could restore hemostatic stability.</p><p><b>METHODS</b>A total of 103 patients (48 females and 55 males; age, 19 - 58 years) were included. Plasma samples were collected before and after treatment as well as after complete remission. Ann II and plasminogen activation were measured in leukemic cells during treatment with 1 micromol/L As2O3.</p><p><b>RESULTS</b>Before As2O3 treatment, Ann II mRNA expression (real-time PCR) was the highest in M3 cells (P < 0.05), higher in M5 cells than that in M1, M2, M4, and M6 cells (P < 0.001), and positively correlated with Ann II protein expression (flow cytometry) (r = 0.752, P < 0.01). Exposure for up to 120 hours to As2O3 (1 micromol/L) had no significant effect on Ann II protein in M1 and M2 leukemic cells, but decreased Ann II protein expression twofold within 48 hours of exposure in M3 cells (P < 0.05) and twofold within 96 hours in M5 cells (P < 0.05). The rate of plasmin generation was higher in APL, M5, and M4 cells than in M1, M2, and M6 cells.</p><p><b>CONCLUSIONS</b>As2O3 may reduce hyperfibrinolysis in AML by downregulation of Ann II. Furthermore, As2O3 affects more than one form of AML (APL, M4 and M5), suggesting its potential role in their management.</p>
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Adult , Female , Humans , Male , Middle Aged , Young Adult , Annexin A2 , Metabolism , Arsenicals , Pharmacology , Bone Marrow Cells , Cell Biology , Metabolism , Cell Survival , Cells, Cultured , Down-Regulation , Leukemia, Promyelocytic, Acute , Metabolism , Oxides , Pharmacology , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
<p><b>OBJECTIVE</b>To trace magnetically labeled MSCs transplanted into the rat livers by magnetic resonance imaging (MRI).</p><p><b>METHODS</b>Feridex and DAPI labeled rat mesenchymal stem cells (MSCs) were injected via portal veins into carbon tetrachloride treated rats. MRI was performed with a clinical 1.5 T MRI machine immediately before the MSCs injection and at h 1, d 3, d 7, and d 14 after the injection, and then the signal-to-noise ratio (SNR) was measured. MRI findings were compared with the liver histopathologies after the slides were stained with fluorescence dye and Prussian blue.</p><p><b>RESULTS</b>The SNR for liver was 1.10+/-0.26 at hour 1, 8.18+/-1.55 at day 3, 11.08+/-1.30 at day 7, and 14.15+/-1.02 at day 14 respectively. Within 7 days after the MSCs transplantation, the SNRs of the livers were significantly lower than those before the transplantation (P less than 0.05). Histologically, the blue fluorescent particles under the fluorescence microscopy matched in distribution with the iron particles on the Prussian blue stained slides.</p><p><b>CONCLUSION</b>The magnetically labeled MSCs transplanted into livers give rise to an obvious signal decrease, and can be tracked with a 1.5 T clinical MRI machine for up to 7 days after MSCs transplantation.</p>
Subject(s)
Animals , Male , Rats , Image Enhancement , Methods , Liver , Pathology , Magnetic Resonance Imaging , Mesenchymal Stem Cell Transplantation , Radioactive Tracers , Rats, WistarABSTRACT
<p><b>OBJECTIVE</b>Mechanical ventilation support is a very important method for the salvage of serious patients. However, it can result in the formation of an adherent matrix of bacteria on the surfaces of implanted materials which is termed "biofilm". Biofilm is dense bacterial communities attached to a solid surface and surrounded by an exopolysaccharide matrix. One of the most important features of bacterial biofilm is their resistance to antimicrobial agents and host immune system components. As a consequence, diseases involving biofilm are generally chronic and difficult to treat. The present study was conducted to explore the relationship between ETT-biofilm and the lower respiratory infection by observing microbial colonization and associated biofilm accumulation on the surface of endotracheal tubes (ETTs) removed from neonates treated with intubated mechanical ventilation.</p><p><b>METHODS</b>Twenty neonates undergoing mechanical ventilation (from January to June in 2005) were recruited into this study. Clinical data about lower respiratory infection for each case were collected. ETTs were collected at the first time of extubation. A sterile control tube was also processed. For each ETT, a 1-cm-long cross-sectional segment was divided into two portions for both scanning electron microscopy (SEM) and aerobic/anaerobic cultures. The presence of biofilm on the surface of ETTs were examined by SEM, meanwhile, bacteria harvested from the surface of ETTs and the secretions of lower respiratory tract were isolated, identified and assessed on antimicrobial susceptibility, respectively.</p><p><b>RESULTS</b>The diagnosis on admission of the twenty cases included: neonatal respiratory distress syndrome (10), meconium aspirate syndrome (2), severe asphyxia (2), pneumatothorax (2), severe pneumonia (1), scleredema neonatorum (1), inborn pulmonary hypoplasia (1) and recurrent apnea (1). Thirteen cases did not present symptoms and signs of lower respiratory infection before mechanical ventilation. However, during the mechanical ventilation process, symptoms and signs of lower respiratory infection presented and lasted until extubation. Nine of the above mentioned thirteen cases (70%) had the same duration of tube use as mechanical ventilation duration (mean: 3.6 days). Observation by SEM showed that colonization was time dependent and the incidence of microbial colonization increased when the duration of tube use exceeded one days (12/20). There were no obvious bacterial colonies except that some amorphous material was noted in 5 of 20 ETTs as early as one day of tube use. Up to 2 days of tube use (4/20), attached bacterial colonization was seen embedded in amorphous material (3/4). Up to 3 days (7/20), a layer of biofilm formation presented on ETTs (5/7). Furthermore, biofilm architecture became more mature and complex if the duration exceeded 3 days. Neither bacteria nor biofilm formation was seen on the control ETT. The results of aerobic/anaerobic cultures showed that there were 14 cultures from ETTs (normal flora grew in 4) and 7 pathogens were isolated; 13 cultures from the secretions of lower respiratory tract (normal flora grew in 1) and 10 pathogens were isolated. Seven samples had the same pathogen both on the surface of ETTs and in the secretions of lower respiratory tract, which accounted for 50% of the positive cultures from ETTs, including Xanthomonas maltophilia (2), Klebsiella pneumoniae (2), Acinetobacter lwoffii (1), Acinetobacter baumannii (1) and normal flora (1). The gram-negative bacteria isolated from the surface of ETTs and the secretions of lower respiratory tract presented multi-resistance to antibiotics.</p><p><b>CONCLUSIONS</b>The ETT-biofilm develops into mature and complex form with the duration of tube use increase. This study provides evidence that there is correlation between microbial colonization, biofilm formation on the surface of ETTs and the lower respiratory infection in neonates who were intubated and ventilated for a prolonged period. ETT-Biofilm could also be a possible source of the recurrent infection. Increased attention must be paid to modification of the ETT to prevent or substantially reduce biofilm formation.</p>
Subject(s)
Female , Humans , Infant , Infant, Newborn , Male , Acinetobacter baumannii , Anti-Bacterial Agents , Therapeutic Uses , Biofilms , Colony Count, Microbial , Equipment Contamination , Gram-Negative Bacteria , Intubation, Intratracheal , Microscopy, Electron, Scanning , Methods , Pediatrics , Pneumonia , Drug Therapy , Respiration, Artificial , Respiratory Tract Infections , Drug Therapy , Trachea , MicrobiologyABSTRACT
The aim of this study was to investigate the gene expression profiles of acute promyelocytic leukemia (APL) cell line NB4 treated with arsenic trioxide (As2O3) by using cDNA microarray. cDNA probes were prepared through reverse transcription from mRNA of NB4 cells treated with or without arsenic trioxide. The probes were labeled with Cy3 and Cy5 fluorescence dyes individually, hybridized with cDNA microarray representing 201 different human genes, and their fluorescent intensities were scanned. The genes were screened through the analysis of the difference in the gene expression profile. The results showed that after the treatment of arsenic trioxide (2 micromol/L), 6 genes were up-regulated, and 12 genes related to apoptosis and signal transduction were down-regulated. The p21, survivin, cdc2 and Wee1Hu genes may be related to the differentiation and/or apoptosis of NB4 cells induced by As2O3. It is concluded that p21, survivin, cdc2 and Wee1Hu may play an important role in the mechanism underling arsenic trioxide-mediated NB4 cell apoptosis.