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ObjectiveTo observe the effect of Hedysari Radix polysaccharide （HRP） on the Janus kinase 2 （JAK2）/signal transducer and activator of transcription protein 3 （STAT3） signaling pathway in diabetic nephropathy db/db mice. MethodFifty db/db mice were randomly divided into model group， irbesartan group （irbesartan suspension， 22.75 mg·kg-1）， and high-， medium-， and low-dose HRP groups （HRP suspension， 200， 100， 50 mg·kg-1） according to the body weight， with 10 mice in each group. Another 10 C57BL/6 mice were assigned to the normal group. The mice were treated with corresponding drugs by gavage， while those in the normal group and the model group received distilled water at 5 mL·kg-1. The mice in the six groups were administered once a day by gavage for 12 consecutive weeks. The uric acid （UA）， triglycerides （TG）， and total cholesterol （TC） were detected. Periodic acid-Schiff （PAS） staining and Masson staining were used to observe the pathological changes in kidney tissues. Western blot and Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） were used to detect the protein and mRNA expression levels of JAK2， STAT3， suppressor of cytokine signaling 3 （SOCS3）， and tumor necrosis factor-α （TNF-α） in the kidney. ResultAfter 12 weeks of treatment， compared with the normal group， the model group showed significant pathological ultrastructural changes in kidney tissues and increased UA， TG， and TC levels （P<0.01）. Compared with the model group， the high- and medium-dose HRP groups and the irbesartan group showed improvement in pathological ultrastructure of kidney tissues and reduced UA， TG， and TC levels （P<0.05， P<0.01）. Compared with the normal group， the model group showed a decrease in SOCS3 protein and mRNA expression levels and an increase in JAK2， STAT3， and TNF-α protein and mRNA expression levels （P<0.01）. Compared with the model group， the high- and medium-dose HRP groups and the irbesartan group showed an increase in SOCS3 protein and mRNA expression levels and a decrease in JAK2， STAT3， and TNF-α protein and mRNA expression levels （P<0.05， P<0.01）. ConclusionHRP can alleviate renal damage in diabetic nephropathy to a certain extent， and its mechanism may be related to the inhibition of the activation of the JAK2/STAT3 signaling pathway.
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Objective:To investigate the mechanism by which chronic psychological stress aggravates intestinal barrier damage and promotes the development of enteritis through inhibiting Wnt/β-catenin pathway, so as to provide a new therapeutic strategy for the clinical diagnosis and treatment of inflammatory bowel disease (IBD).Methods:A comorbidity model of chronic psychological stress and enteritis was established using C57BL/6J mice. HE staining was used to analyze the effects of chronic psychological stress on the intestinal pathological damage in mice with enteritis. ELISA was used to detect the expression of proinflammatory cytokines. The ultrastructural changes of colonic cells and the state of intestinal mucus layer were observed under transmission electron microscope and scanning electron microscope. The secretion of mucoprotein 2 (MUC2) and the expression of cell proliferation marker Ki67 were detected by immunofluo rescence staining. The numbers of goblet cells were detected by Alcian blue-periodic acid-Schiff (AB-PAS) staining. Western blot was performed to analyze the expression of tight junction protein between intestinal epithelial cells, β-catenin which was a key protein of Wnt/β-catenin pathway maintaining crypt proliferation, and downstream protein c-myc.Results:The sugar water consumption ratio decreased, but tail suspension immobility time, the swimming immobility time and the expression of corticotropin releasing hormone (CRH) in hypothalamus increased (all P<0.05) in the stress group as compared with those in the control group. Chronic psychological stress promoted weight loss and colonic shortening in mice with enteritis, exacerbated pathological damage and enhanced the release of pro-inflammatory factors. Moreover, increased disappearance of intestinal epithelial microvilli and severe cellular ultrastructural damage were also observed in the stress+ dextran sulfate sodium salt (DSS) group. There was no pathological damage in the control and stress groups. Chronic psychological stress aggravated intestinal barrier injury and inhibited intestinal barrier repair by inhibiting Wnt/β-catenin pathway. Conclusions:In the mouse model of DSS-induced enteritis, chronic psychological stress preconditioning inhibited the Wnt/β-catenin pathway, weakened the repair ability of intestinal epithelium, aggravated the loss of mucus layer of intestinal barrier and the damage of tight junction structure, and promoted the development of enteritis. In the absence of enteritis, chronic psychological stress had no significant effects on the Wnt/β-catenin pathway and the intestinal barrier.
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Objective:To investigate the association of XPD rs13181 (codon751A/C, Lys751Gln), rs238406 (codon156C/A, Arg156Arg), XPC rs2279017 (i11C/A), and XRCC4 rs3734091 (codon247T/C, Ala247Ser) polymorphisms with colorectal cancer (CRC) susceptibility. Methods:A total of 338 patients with CRC who were treated at the Beijing Cancer Hospital from April 2013 to January 2016 (case group) and 315 healthy controls (control group) were genotyped using TaqMan technology. Results:The genotype GT and G alleles of XPD rs13181 increased the risk of CRC (GT>TT, adjusted OR=1.69, 95%CI=1.15-2.47, P=0.007;G>T, adjusted OR=1.77, 95%CI=1.19-2.64, P=0.005). The genotype GT and T alleles of XRCC4 rs3734091 increased the susceptibility of CRC (GT>GG, adjusted OR=9.02, 95%CI=5.61-14.50, PG, adjusted OR=4.06, 95%CI=2.49-6.61, P<0.001). Analyses of XPD rs13181 and rs238406 indicated that the haplotype GT significantly decreased the risk of CRC (adjusted OR=0.39, 95%CI=0.18-0.85, P=0.018). Moreover, the combinations of the XPC rs2279017 G allele and the XRCC4 rs3734091 T allele (adjusted OR=28.43, 95%CI=6.85-117.95, P<0.001) and the XPD rs13181 G allele and the XRCC4 rs3734091 T allele (adjusted OR=10.24, 95%CI=4.69-22.35, P<0.001) exhibited significantly increased CRC risk. Conclusion:The polymorphisms of XPD rs13181 and XRCC4 rs3734091 increased the risk of CRC.
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Objective To observe the clinical efficacy of acupuncture and miuci therapy combined with Qinggan Tongqiao Decoction for sudden sensor-neural hearing loss (SSHL) and the effects on level of plasma endothelin (ET) and serum nitrate oxide (NO). Methods Totally 110 patients with SSHL were randomly divided into treatment group and control group, with 55 patients in each group. The control group was given Qinggan Tongqiao Decoction combined with acupuncture routine treatment, and treatment group was given miuci therapy on the basis of the control group, two weeks as a course of two groups. Level changes of plasma ET and serum NO, and pure tone listening threshold changes before and after treatment were observed. Tinnitus and hearing efficacy was evaluated. Results The total effective rate for tinnitus efficacy was 94.54% (52/55) in the treatment group and 85.45% (47/55) in the control group, with statistical significance (u=2.038, P=0.040). The total effective rate for hearing efficacy was 96.46%(53/55) in the treatment group and 81.82% (45/55) in the control group, with statistical significance (u=2.913, P=0.040). Compared with before treatment, the level of plasma ET decreased and the level of serum NO increased after the treatment (P<0.05); After the treatment, the level of plasma ET in the treatment was lower than the control group, and the level of serum NO in the treatment group was higher than the control group (P<0.05). Pure tone threshold scores in the two groups on treatment 3, 7, 14 d were significantly improved (P<0.05), and pure tone threshold scores in the treatment group were significantly lower than the control group (P<0.05). Conclusion Acupuncture and miuci therapy combined with Qinggan Tongqiao Decoction can significantly reduce clinical symptoms and enhance the hearing level of SSHL patients. The mechanism mainly lies in regulating levels of plasma ET and serum NO, and improving the inner ear micro-circulation.
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Parathyroid hormone ( PTH) is an important hormone secreted by parathyroid cells , and regulates the metabolism of calcium and phosphorus in the body .In recent years , the toxic effect of PTH on myocardium has been re-ported.Calcium-sensing receptor (CaSR), a member of G protein-coupled receptor family, can feel the subtle change of extracellular calcium concentration and regulate intracellular calcium concentration through multifarious ways in order to control the secretion of PTH .The expression of CaSR is observed in parathyroid cells , renal tubular epithelial cells , myo-cardial cells, etc.Intracellular calcium, as a second messenger, participates in various cell functions , such as excitation-contraction coupling , fertilization and so on .The injury of myocardial cells is intimately linked with high concentrations of PTH and intracellular calcium , and high expression of CaSR .
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AIM: To study the effect of calcium sensing receptor (CaSR) on icariin (ICA) induced mouse embryonic stem cells ( mESCs) to differentiate into cardiomyocytes in vitro.METHODS:mESCs were cultured to embry-oid bodies ( EBs) by direct suspension method and the differentiation of EBs into cardiomyocytes was induced by ICA.The expression of cardiac specific proteinsα-actinin and cardiac troponin-I ( cTnI) was analyzed by Western blot and immuno-fluorescence.The differentiation rate was determined by flow cytometry.The ultrastructure of the derived cardiomyocytes was further characterized by transmission electron microscopy.The expression of cardiac-specific transcription factors Nkx2.5 and GATA-4,as well as CaSR was detected by Western blot.RESULTS: After induction with ICA, the positive characteristics of myocardial cells appeared in the EBs cultured for 2 d.The expression of cardiac-specific sarcomeric pro-tein actinin (α-actinin) and cTnI showed an overall upward trend by Western blot in different phases of ICA induced differ-entiation.The expression of CaSR, Nkx2.5 and GATA-4 was the highest at an early stage of ICA-induced differentiation. Neomycin (an activator of CaSR) up-regulated CaSR, NKx2.5 and GATA-4 expression in the EBs at early stage of ICA-in-duced differentiation, all of which were reversed by NPS2390 ( an inhibitor of CaSR) .CONCLUSION:CaSR is function-ally expressed in mESC-derived cardiomyocytes, and activation of CaSR is involved in the differentiation of mESCs into car-diomyocytes by facilitating the expression of NKx2.5 and GATA-4.
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AIM: Ischemic post-conditioning ( PC) plays an important role in cardioprotection from ischemia /reperfusion ( I/R) injury in the young heart but not in the aging hearts .The physiological and pathological roles of hydrogen sulfide ( H2 S) in the regulation of cardio-vascular functions have been recognized .Whether H2 S is involved in the recovery of PC-induced cardioprotection in the aging hearts is unclear.METHOD:The male Wistar young rats (3-month-old), the aging rats (24-month-old), primary cultured cardiomyocytes and the aging cardiomyocytes induced by D-galactose suffered from I/R (or H/R) and PC.RESULTS:I/R (or H/R) decreased H2S production rate and cystathionine γ-lyase (CSE) expression, aggravated cardiomyocyte damage , apoptosis, myocardial infarct size and oxidative stress, reduced cardiac function, increased the levels of Bcl-2, cleaved caspase-3 and caspase-9 mRNA and proteins, promo-ted the release of cytochrome c and mPTP opening, down-regulated the phosphorylation of ERK 1/2, PI3K, Akt and GSK-3βand mito-chondrial membrane potential , up-regulated the phosphorylation of IκBα, NF-κB, JNK2 and STAT3, and inhibited PKC-ε transloca-tion and mitochondrial ATP-sensitive K channels (mitoKATP) in isolated young and aging hearts as well as normal and aging cardiomyo-cytes.PC suppressed myocardial I/R injury in the young heart but not in the aging hearts .Supply of NaHS not only increased PC-in-duced cardioprotection in the young hearts and cardiomyocytes , but also attenuated I/R injury and significantly recovered the cardiopro-tective role of PC in the aging hearts and cardiomyocytes .CONCLUSION:The exogenous H 2 S restores PC-induced cardioprotection through inhibiting oxidative stress via the down-regulation of NF-κB and JAK2-STAT3 pathways and mPTP opening by the up-regulation of ERK1/2-GSK-3β, PI3K-Akt-GSK-3βand PKC-ε-mitoKATP pathways in the aging hearts and cardiomyocytes .These findings provide a novel potential target for the treatment of aging ischemic cardiomyopathy .
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AIM:To investigate the relationship between polyamine metabolism and hypoxia /ischemia ( H/I)-induced cell apoptosis and to determine the mechanisms by which exogenous spermine protects cell apoptosis against AMI in rats .METHOD:The left anterior de-scending coronary artery ( LAD) of the Wistar rats were ligated , and neonatal rat cardiomyocytes were placed under hypoxic conditions for 24 h to establish the model of AMI (or H/I).Exogenous spermine was administered by intraperitoneal injection (2.5 mg/kg daily for 7 days) in vitro and subjected to the cell medium at 5μmol/L as a pre-treatment therapy.RESULTS:AMI (or H/I) induced an increase in polyamine catabolized enzyme SSAT and a decrease in polyamine biosynthesis enzyme ODC , which result in endogenous spermine and spermidine decrease and putrescine increase .At the same time, AMI ( or H/I) lowered cardiac function , increased cTnI and CK-MB concentrations , aggravated myocardial infarct size , cardiomyocyte damage and apoptosis , raised ROS generation , increased the expression of cleaved caspase-3, cleaved caspase-9 and endoplasmic reticulum stress (ERS)-related proteins, promoted the release of cytochrome C and mPTP opening , down-regulated Bcl-2 expression and the phosphorylation of ERK 1/2, PI3K, Akt and GSK-3β, and activated PERK and eIF 2αphosphorylation .Spermine pre-treatment reversed the above-motioned changes .CONCLUSION:AMI ( or H/I ) could induce cardiomyocyte apoptosis and polyamine metabolism disorder .Exogenous spermine attenuates cardiac injury through scavenging the ROS and inhibiting mPTP opening and ERS injury .These findings provide a novel target for the prevention of apoptosis in the setting of AMI .
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Huntington's disease (HD) is an autosomal dominant neurodegenerative disease characterized by a triad of progressive motor dysfunction, cognitive decline and psychiatric disturbances. The hallmark of HD is the distinctive choreiform movement disorder that typically has a subtle, insidious onset in the fourth to fifth decade of life and gradually worsens over 10 to 20 years until death. Notably, two-thirds of HD patients present with chorea and one third with mental changes. The prevalence of psychiatric symptoms is significantly higher than in the general population, and is estimated to be around 66–73%. Here, we report a unique case of subsequent onset of HD in a patient previously treated for schizophrenia and complicated by the extrapyramidal side effects to antipsychotics.
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Humans , Antipsychotic Agents , Chorea , Huntington Disease , Neurodegenerative Diseases , Prevalence , Psychotic Disorders , SchizophreniaABSTRACT
<p><b>OBJECTIVE</b>The aim of this study was to investigate the expression of microRNA-100 (miR-100) and its relation with prognosis in colorectal cancer (CRC).</p><p><b>METHODS</b>The expression of miR-100 was analyzed by quantitative real-time PCR (qRT-PCR) in 172 CRC tissue samples. The relation of miR-100 expression patterns with clinical pathological significance in CRC was analyzed. The effects of alterations of miR-100 expression and its consequences on CRC cell proliferation, apoptosis and migration were demonstrated in cells cultured in vitro.</p><p><b>RESULTS</b>The relative expression of miR-100 in CRC tissues and peritumoral tissues were -6.185 ± 1.921 and -3.698 ± 1.786, respectively, with a significant difference between the two groups (P<0.01). There was a significant difference between the relative expression of miR-100 in CRC with lymph node metastasis (-5.706 ± 1.809) and without lymph node metastasis (-6.775 ± 1.902, P<0.01). The relative expression of miR-100 in tumors of different TNM stages were -7.267 ± 1.888 in stage I, -6.443 ± 1.859 in stage II, -5.923 ± 1.796 in stage III, and -4.639 ± 1.516 in stage IV, with a significant difference among them (P<0.01). Different differentiation grades showed different expression of miR-100, i.e. -7.389 ± 1.828 in well differentiated tumors, -6.095 ± 1.843 in moderately differentiated tumors, and -5.476 ± 2.088 in poorly differentiated tumors (P<0.01). There was no significant correlation between miR-100 expression and overall survival rates of the CRC patients (P=0.179). Overexpression of miR-100 in the CRC cell line HCT-8 inhibited cell proliferation, but promoted cell apoptosis and migration.</p><p><b>CONCLUSIONS</b>The expression of miR-100 is correlated with lymph node metastasis, TNM stage and differentiation grade, and may be a potential biomarker indicating the development of CRC.</p>
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Humans , Apoptosis , Cell Differentiation , Cell Proliferation , Colorectal Neoplasms , Metabolism , Mortality , Pathology , Gene Expression Regulation, Neoplastic , Lymphatic Metastasis , MicroRNAs , Metabolism , Neoplasm Staging , Prognosis , Real-Time Polymerase Chain ReactionABSTRACT
Aim To study the effects of difluoromethy-lornithine (DFMO)on cardiomyocytes hypertrophy and apoptosis induced by isoproterenol (ISO).Methods The cardiomyocytes were divided into four groups ran-domly:Control group,ISO group,ISO+DFMO group and (ISO +DFMO +Putrescine)group.The hyper-trophic model of cardiomyocytes was induced by ISO, the cardiomyocytes were pretreated with 0.5 mmol · L-1 DFMO and 0.5 mmol·L-1 putrine,the gene ex-pression of ANP,the surface area of cardiomyocytes and the contents of LDH and MDA were measured. Apoptotic value of cardiomyocytes was observed by An-nexin V/PI,the gene and protein expressions of Bcl-2 and Bax were detected by real-time PCR and Western blot.Results Compared with ISO group,the cell sur-face area,the gene level of ANP,the apoptosis value of cardiomyocytes and the contents of LDH and MDA were decreased in ISO +DFMO group (P <0.05 ). Meanwhile,DFMO pretreatment upregulated the gene and protein expressions of Bcl-2 (P <0.05 ), in-creased the ratio of Bcl-2/Bax (P<0.05 ),downregu-lated the gene and protein levels of Bax (P<0.05 ). Conclusion The cardiomyocyte hypertrophy and ap-optosis induced by ISO can be protected by DFMO pre-treatment,which may be associated with the expression of apoptotic gene Bcl-2 and Bax.
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AIM:To observe the functional expression of calcium-sensing receptor (CaSR) in the mouse em-bryonic stem cells (mESCs).METHODS:The expression and distribution of CaSR were detected by Western blotting and immunofluorescence observation in 129 mouse ES-D3 cells.The intracellular concentration of free calcium ([Ca2+]i) was determined by confocal laser scanning microscopy .The cell viability was analyzed by MTT assay and flow cytometry .RE-SULTS:CaSR protein was expressed in mESCs .Extracellular calcium or neomycin significantly increased the expression of CaSR and [Ca2+]i.Neomycin increased the cell viability , up-regulated the protein expression of p-ERK2.These effects of neomycin were inhibited by NPS2390.CONCLUSION:CaSR is expressed in mESCs .The activation of CaSR is involved in the proliferation of mESCs .
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Objective To explore the diagnostic consistency and correlation between MR discography (MRD) and CT discography (CTD) in diagnosing chronic low back pain. Methods Guided by C - arm fluoroscopy the mixed solution of gadoterate meglumine (GD-DOTA) and Iohexol (GD-DOTA at a dilution of 1 ∶ 400 with Iohexol) was injected into 96 lumbar intervertebral discs of the 36 patients. CT scanning was performed at 15 minutes after the injection of contrast, and axial together with sagittal SE T1WI MR scanning was carried out one hour after the injection. CTD and MRD images were randomly numbered and were independently evaluated by two experienced radiologists according to Dallas discogram scale in order to assess the diagnostic consistency and correlation between (MRD) and (CTD). In addition the diagnostic value of MRD was evaluated. Results The results revealed that in determining disc degeneration grade CTD and MRD were highly consistent with each other(Kappa = 0.836, P < 0.01), and the diagnostic results judged by the two reviewers were essentially in agreement (ICC = 1.00, P < 0.01; r = 0.997, P < 0.01). Higher consistency (Kappa = 0.836, P < 0.01) and correlation(ICC = 0.90, P < 0.01; r = 0.869, P < 0.01; Kappa =0.836, P < 0.01) in determining annulus rupture extent were also obtained. Conclusion MRD is an accurate diagnostic method for the determination of disc degeneration and the severity of annulus rupture, and this technique has greater consistency and correlation with CTD in diagnosing chronic low back pain.
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<p><b>OBJECTIVE</b>The aim of this study was to identify six miRNAs expressed in plasma of patients with pancreatic cancer (PCa) and analyze their value as a diagnostic index of pancreatic cancer.</p><p><b>METHODS</b>Plasma total RNAs were extracted from 30 PCa patients and 26 normal controls, and the abundance of six microRNAs was measured using real-time PCR. The possibility to combine them with CA19-9 as diagnostic biomarkers was analyzed.</p><p><b>RESULTS</b>The expression level of miR-21, miR-210, miR-155, miR-20a, miR-25 and miR-196a in plasma of patients with pancreatic cancer were 1.65×10(6), 5.98×10(4), 2.83×10(3), 3.47×10(6), 2.76×10(6), and 1.03×10(3) (copies/µl), while the normal controls were 4.08×10(5), 2.54×10(4), 8.55×10(2), 1.79×10(6), 9.32×10(5), and 4.67×10(2) (copies/µl), respectively, with a significant difference between the two groups (P < 0.05). The areas under the ROC curve of miR-21, miR-210, miR-155, miR-20a, miR-25 and miR-196a were 0.893, 0.810, 0.820, 0.766, 0.816 and 0.729, respectively. MiR-21 had the highest diagnostic value when it was used as diagnostic marker alone. The combination of miR-155 and miR-25 was more effective to distinguish PCa from normal than to be used alone, and the area under the ROC curve was 0.913 (95%CI 0.838-0.988) .When CA199 associated with miR -210 and miR-25, respectively, the areas under the ROC curves were 0.96 (95%CI was 0-1.0) and 0.942 (95% CI was 0.876-1.0), which were higher than CA199 alone (0.862, 95%CI was 0.748-0.975). There was a high improvement in diagnostic sensitivity and accuracy when miR-210 and miR-25 were combined with CA19-9, respectively.</p><p><b>CONCLUSIONS</b>Plasma miR-21, miR-155, miR-25, miR-210 have diagnostic value for pancreatic cancer, and deserve further study.</p>
Subject(s)
Aged , Female , Humans , Male , Middle Aged , Biomarkers, Tumor , Blood , Genetics , CA-19-9 Antigen , Blood , Case-Control Studies , MicroRNAs , Blood , Pancreatic Neoplasms , Blood , Diagnosis , Genetics , ROC Curve , Real-Time Polymerase Chain ReactionABSTRACT
<p><b>OBJECTIVE</b>To investigate the controlling effects of bionic glue on Paratrioza sinica.</p><p><b>METHOD</b>P. sinica and bionic glue were chosen as materials to investigate the adhesive rate, mortality rate, and study the effects of behavior of P. sinica and growth of leaves sprayed with bionic glue.</p><p><b>RESULT</b>Spraying of the bionic glue can significantly increase the adhesive rate of P. sinica with no obviously repellency, and it can be used to control the adults of P sinaca in field with no significant effect on the growth of wolfberry leaves.</p><p><b>CONCLUSION</b>Bionic glue has significant controlling effects on adults of P. sinica, it can be used as an eliminator of adults of P. sinica in field at the beginning of the vegetation season to suppress of the development of P. sinica population.</p>
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Animals , Adhesives , Pharmacology , Hemiptera , Insect Control , Insecticides , Pharmacology , Lycium , Parasitology , Plant Diseases , Plant Leaves , ParasitologyABSTRACT
Calcium sensing receptors (CaSR) is a member of super-family of G-protein coupling receptors. This review first introduced the concept, construction features, distribution, functions, decision methods, moderators, genetic locus of CaSR and its relationship with some diseases concisely. Then this article described the investigation progress of CaSR in cardiovascular system intensively, including the expression pattern, role and signal pathways of CaSR in rat myocardium in normal, ischemia-reperfusion injury, apoptosis and cardiac hypertrophy;the role and mechanism of CaSR in calcium homostasis regulation of rat myocardium, endoplasmic reticulum (ER) stress and cardiac ischemic preconditioning and postconditioning. The metabolism rule, physiological significance and pathological action of polyamine in cardiac cells;the increase of CaSR expression in cardiac tissue of artherosclerosic rat and its effect on sensitivity to acute myocardial infarction are also discussed. In the end, the research perspective of CaSR in cardiovascular system was anticipated.
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AIM: To explore the effects and possible mechanism of exogenous spermine on the apoptosis of primary cultured neonatal cardiomyocytes induced by simulated ischemia-reperfusion (I/R) injury. METHODS: To establish a model of simulated I/R, the primary cultured neonatal rat cardiomyocytes were incubated in ischemia-mimetic solution (under the conditions of hypoxia plus serum deprivation) for 2 h, and re-incubated the cells in normal culture medium for 24 h. The apoptotic cell death was assayed by flow cytometry. The morphological alterations of the cells were observed under transmission electron microscope. The transcription and expression of Fas and FasL were determined by the methods of RT-PCR, Western blotting and immunofluorescence. RESULTS: The cells exposed to I/R underwent significant apoptosis, and the percentage of apoptotic cells was 27.4%±1.8%, much higher than that in normal group (5.7%±0.3%). In I/R group the evident histopathological changes were observed and the myocardial transcription and expression of Fas and FasL were significantly upregulated. Compared to normal group, mRNA expression of Fas and FasL increased 2.2 folds and 2.4 folds, respectively, and their proteins increased 1.7 folds and 1.9 folds at 24 h of reperfusion respectively (P<0.01). Pretreatment with 10 μmol/L spermine significantly inhibited apoptosis of I/R injured cells, and the percentage of apoptotic cells was 21.7%±1.3% (P<0.01, as compared to I/P group). Spermine also suppressed the expression of Fas and FasL significantly (P<0.05 or P<0.01, as compared to I/P group). CONCLUSION: Spermine plays anti-apoptotic effect on the cultured neonatal myocardial cells under the condition of I/R injury by suppressing the expression of Fas/FasL.
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In order to explore the environmental pest management method of Dorysthenes hydropicus, three strains of entomopathogenic nematodes, viz. Heterorhabditis bacteriphora (H06), Steinernema scapterisci (SS), S. carpocapsae (All) were used on larvae of Dorysthenes hydropicus, with treatments of 0, 5 000 and 10 000 nematodes each larva. The result showed that these three strains viz. All, H06 and SS had high lethal effects on the larvae. Lethal rates had dose-effects relationship with inoculation amounts. High dose treatments resulted in high mortalities and led to quick death, especially in the treatment of H06. Treatment of H06 with 10 000 nematodes per larva resulted in 100% mortality after inoculated 4 days. Different strains of these nematodes had various lethal characters, H06 with only one peak mortality, the larvae died quickly after inoculated, while All and SS with 2 peak mortalities, there was a stable stage with low mortality between the 2 peak mortalities. Entomopathogenic nematodes could be used as a hopeful method for controlling of Dorysthenes hydropicus in fields.
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Animals , Larva , Parasitology , Moths , Parasitology , Physiology , Nematoda , Physiology , Pest Control, Biological , MethodsABSTRACT
AIM: To explore the role and possible mechanism of polyamine in L - arginine inhibiting cardiac hypertrophy induced by isoproterenol (ISO). METHODS: Hypertrophic model of rats was established using ISO. Pretrea-ted with L - arginine, hypertrophy status of rats was determined by hypertrophy coefficient, collagen content and the expression of ANP mRNA. High performance liquid chromatography ( HPLC) was used to measure the concentrations of polyamines. Western blotting was performed to detect the expressions of ornithine decarboxylase ( ODC) and spermidine/ spermine Nl - acetyltransferase (SSAT). The activity and levels of NOS and NO in serum were also observed. RESULTS : Hypertrophy coefficient and expression of ANP mRNA increased significantly after injection of ISO for 7 d. Moreover, cardiac muscle fibres became thick and disorganized. Pretreated with L - arginine, the above index decreased. Meanwhile , the concentration of polyamine was decreased and plasma NO content and NOS activity were increased, the expression of ODC was downregulated and the expression of SSAT was upregulated. CONCLUSION: Exogenous L - arginine inhibits cardiac hypertrophy through downregulating L - arginine/polyamine pathway and upregulating L - arginine/NO pathway.
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AIM: To explore the protective role of ornithine decarboxylase (ODC)/polyamines system in the myocardium induced by ischemic preconditioning in rats.METHODS: The experiment model of simulating myocardial ischemia-reperfusion was replicated by Langendorff perfused rat heart. The hearts were randomly divided into six groups: control group, ischemic-reperfusion group (IR), weak ischemic preconditioning group (IPCw), strong ischemic preconditioning groups (IPCs) and inhibitor groups (DF-EG-IPCw and DF-EG-IPCs). The expression of ODC was quantified by Western blotting analysis. The contents of polyamines (putrescine, spermidine, spermine) in cardiac tissue were detected with high performance liquid chromatography. The hemodynamics was obtained using the PowerLab 8/SP TM data acquisition system. The infarct size was measured using triphenyltetrazolium chloride (TTC) staining and the apoptosis cardiomyocytes were observed under optic microscope after TUNEL method treatment. RESULTS: In contrast with control group, in IR group the putrescine contents increased, the expression of ODC was down-regulated, the contents of spermine and the total polyamine pool decreased (P<0.05). At the same time, the cardiac function declined, with an increase in myocardium infarct size and the apoptosis rate of cardiomyocytes (P<0.05). When compared with IR group in terms of LVDP, HR and CF, both IPCw and IPCs groups had significant improvements in cardiac functions (P<0.05). These two groups also had smaller myocardium infarct size (P<0.01) and apoptosis rate of cardiomyocytes (P<0.01). When compared with IR group, the expression of ODC, the contents of spermine and the total polyamine pool increased in both IPCw (P<0.05) and IPCs groups (P<0.01), but the putrescine contents declined. In the respective inhibitor groups of the weak and ischemic preconditioning, the expression of ODC and the levels of putrescine, spermidine, spermine and the total polyamine pool decreased remarkably (DF-EG-IPCw vs IPCw, P<0.05; DF-EG-IPCs vs IPCs, P<0.01), while the myocardium infarct size and apoptosis rate of cardiomyocyte were relatively bigger in both inhibitor groups (P<0.05). Also, the heart function decreased significantly in terms of LVDP, HR and CF compared to their matched ischemic preconditioning group (P<0.05).CONCLUSION: Ischemic preconditioning significantly up-regulates the ODC / polyamines system in Langendorff perfused rat hearts and provides protective effects on myocardium with ischemia/reperfusion injury. Inhibition of bio-synthesis of polyamine abolishes the cardiac function improvement and the decreases the infarct size and apoptosis rate induced by ischemic preconditioning. It reveals that the ornithine decarboxylase (ODC) /polyamines system may be involved in the protection of myocardium induced by IPC in rats.