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Objective To investigate the levels of 5-hydroxymethylcytosine (5-hmC) and teneleven-translocation proteins (TET1/2/3) in diagnosis and prognosis prediction of hepatocellular carcinoma (HCC).Methods The expression of 5-hmC in 130 cases of HCC tissues were detected by immunohistochemical staining.Kaplan-Meier test was used for survival analysis.TET family plays critical role in the conversion of 5-methylcytosine (5-mC) to 5-hmC.The TET levels were detected by using immunohistochemical staining and RT-PCR,the correlation between 5-hmC and TET was analyzed.Results The level of 5-hmC decreased in HCC tissues,as compared with non-tumor tissues,the expression of TET1 was downregulated in HCC.There was significant difference in the expression between low and high grades of HCC tissues (x2 =10.611,P =0.001).Kaplan-Meier survival analysis showed that there was significant difference between the 5-hmC expression level and the survival rate of HCC patients (x2 =4.412,P =0.036).Conclusions In HCC tissues the expression of 5-hmC was specifically downregulated.Low 5-hmC level is significantly correlated with poor differentiation of the tumor and worse overall survival.Decreased expression of TET1 is likely one of the mechanisms underlying 5-hmC loss in HCC.
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Objective@#To establish to paraquat poisoning acute lung injury animal model to study the therapeutic effect of Salvia polyphenols acid salt of paraquat-induced acute lung injury.@*Methods@#Adult male Wister rats 120, were randomly divided into three groups: the paraquat exposure group, the start of the experiment to give a one-time 20% paraquat dope orally 50 mg/kg body weight of rats; salvianolate treatment group, the start of the experiment paraquat to give a one-time 20% the stock solution orally 50 mg/kg body weight of rats, and then given daily intraperitoneal injection of 200 mg/kg body weight of rats salvianolate; blank control group was given the same amount normal saline. The exposure group, the treatment group and control group rats were sacrificed after anesthesia in the 3rd, 7th, 14th, 21st day from the beginning of the experiment respectively, and taken out and preserved venous blood specimens and lung tissue to be tested. Venous detection heme oxygenase-1 (HO-1) , the lung tissue detection heme oxygenase-1 (HO-1) , hydroxyproline (HYP) . And do biopsy specimens from some of the lung tissue, HE and Masson staining observed by optical microscope.@*Results@#Compared with control group, model group 7, 14, 21 days had elevated levels of serum and lung tissue HO-1 (all P<0.05) ; Treatment group 3, 7, 14, 21 days increased (all P<0.05) , and 3, 7, 14 days is higher than the model group (compared with model group, P<0.05) . Compared with control group, treatment group 3 days and model group, 14, 21 days HYP content in lung tissue increased significantly (all P<0.05) ; 21 days, compared with model group, HYP content of treatment group reduce obviously, the difference was statistically significant (P<0.05) . Optical microscope observation, lung tissue damage and aggravated with the experimental increase in the number of days, inflammatory cell infiltration, alveolar septal fibrosis gradually formed. The treatment group experimental animal lung tissue to reduce inflammation, lung fibrosis relief.@*Conclusion@#Paraquat (50 mg/kg body weight) to fill the stomach can be induced model of acute lung injury in the rats. The serum HO-1 expression and HO-1, HYP content in lung tissue increased obviously in model rats. Salvia miltiorrhiza polyphenols acid salt to a certain degree and stage influenced the expression of HO-1 and HYP, relieve acute lung injury and pulmonary fibrosis, has certain curative effect in the treatment of paraquat poisoning.
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Introducción: la hiperplasia suprarrenal congénita engloba todos los trastornos hereditarios de la esteroidogenia suprarrenal del cortisol, trasmitido por las mutaciones con carácter autosómico recesivo. El déficit de enzimático 21 hidroxilasa es la forma más frecuente de esta enfermedad, constituyendo del 90 al 95% de los casos. Objetivo: describir la correlación fenotipo-genotipo en pacientes con hiperplasia suprarrenal congénita diagnosticados por tamizaje neonatal. Material y método: se realizó un estudio descriptivo, retrospectivo, a 8 neonatos con hiperplasia suprarrenal congénita por déficit de 21 hidrixilasa diagnosticado por tamizaje. Se les pidió consentimiento informado, para realizar el examen físico y la extracción de sangre para cuantificación de 17 hidroxiprogesterona y estudio molecular. Este último se efectuó en el Centro Nacional de Genética; se buscaron las mutaciones (P30L, Intrón 2, deleción de 8 pb y G318X). Resultados: el 62,5% de los pacientes presentaron síntomas clínicos, los dos pacientes en los que se presentó la forma clásica perdedora de sal, que es la forma neonatal grave, exhiben varias mutaciones en el seudogen y en el gen activo la mutación del intrón 2, el 50% de las madres fueron homocigóticas o heterocigóticas a estas mutaciones. Conclusiones: la mutación más frecuente encontrada fue la del Intrón 2. Entre las características clínicas, prevaleció la macrogenitosomía y virilización simple. Se logró realizar la correlación del fenotipo y el genotipo a la mayoría de los afectados.
Introduction: congenital adrenal hyperplasia encompasses all inherited disorders of adrenal steroidogenesis cortisol, transmitted by autosomal recessive mutations. The enzyme 21-hydroxylase deficiency is the most frequent form of this disease, constituting 90 to 95% of cases. Objective: To correlate the phenotypic characteristics with genotype in patients suffering from congenital adrenal hyperplasia diagnosed by neonatal screening in Holguin province, Cuba. Material and method: a retrospective descriptive study was performed in 8 neonates with congenital adrenal hyperplasia per 21-hydroxylase deficiency, diagnosed by screening. They were asked for informed consent to perform physical examination and blood collection for the quantification of 17-hydroxyprogesterone and a molecular study that was performed at the Genetic National Center; mutations (P30L, Intron 2, 8 bp deletion and G318X) were sought. Results: 62.5% of the patients presented clinical symptoms, the two patients who presented the classic salt loss form, which is the severe neonatal disorder, exhibit various mutations in the pseudogene and the active gene mutation Intron 2, where 50% of mothers were homozygous or heterozygous for these mutations. Conclusions: the most frequent mutation found was Intron 2. Among the clinical features, macrogenitosomia prevailed and simple virilization. The correlation of phenotype-genotype was accomplished to the most affected.
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BACKGROUND: Reperfusion in ischemia is believed to generate cytotoxic oxidative stress, which mediates reperfusion injury. These stress conditions can initiate lipid peroxidation and damage to proteins, as well as promote DNA strand breaks. As biliverdin and bilirubin produced by heme oxygenase isoform 1 (HO-1) have antioxidant properties, the production of both antioxidants by HO-1 may help increase the resistance of the ischemic brain to oxidative stress. In the present study, the survival effect of HO-1 was confirmed using hemin. METHODS: To confirm the roles of HO-1, carbon monoxide, and cyclic guanosine monophosphate further in the antioxidant effect of HO-1 and bilirubin, cells were treated with cycloheximide, desferoxamine, and zinc deuteroporphyrin IX 2,4 bis glycol, respectively. RESULTS: HO-1 itself acted as an antioxidant. Furthermore, iron, rather than carbon monoxide, was involved in the HO-1-mediated survival effect. HO-1 activity was also important in providing bilirubin as an antioxidant. CONCLUSION: Our results suggested that HO-1 helped to increase the resistance of the ischemic brain to oxidative stress.
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Animals , Rats , Antioxidants , Bilirubin , Biliverdine , Brain , Carbon Monoxide , Cycloheximide , DNA , Guanosine Monophosphate , Heme , Heme Oxygenase (Decyclizing) , Hemin , Iron , Ischemia , Lipid Peroxidation , Microvessels , Oxidative Stress , Oxygen , Oxygenases , Reperfusion , Reperfusion Injury , ZincABSTRACT
Objective To investigate the expression and distribution of COX-2 in benign and malignant breast disease,and the relationship among COX-2 expression and the clinicopathologic factors,hormone receptors.Methods Immunohistochemistry Was used to investigate the expression and distribution of COX-2 in 8 accessory.breasts,31 BPBDs(15 in adnosis,16 in fibroadnoma),70 IDCs(35 of them accompany with DCIS);The relationships between the expression of COX-2 and hormone receptor(ER and PR),C-erbB-2.and the clinicopathologic factors were studied respectively.Results①The expression of COX-2 in various kinds mammary diseases:8 accessory breasts were all negative;the positive rate of COX-2 in BPBD was 96.5%(30/31),among these,the positive rate of COX-2 in adnosis was 93.3%(14/15),and in fibroadnonm was 100%(16/16).The positive rate of COX-2 in DCIS was 85.7%(30/35).The intensity of COX-2 expression in DCIS of periph-IDC was higher than that in corresponding carcinoma.The positive rate of COX-2 in IDC Was 84.3%(59/70),the expression of COX-2 Was different among non-cancer gland.②COX-2 expression in BPBD was obviously higher compared with COX-2 in IDC,the difference was significant in statistics(χ2=9.39,P＜0.025).③The expression of COX-2 in DCIS was correlative with low histological differentiation(χ2=10.98,P＜0.005),PR negative(P=0.019,Fisher exact probability),and C-erbB-2 positive(P=0.0008,Fisher exact probability).The expression of COX-2 in IDC was correlative with lymph node metastasis(χ2=4.09,P＜0.05),tissue poorly differentiated(P=0.004,Fisher exact probability),PR negative(χ2=6.91,P＜0.01)and C-erbB-2 positive(χ2=5.94,P＜0.025).Conclusion COX-2 does not express in normal mammary gland.It is high-expressed in BPBD,DCIS and IDC;These indicate the up-regulation of COX-2 expression not only participates in breast carcinogenesis,but also promotes its metastasis and progression,and suggests COX-2 may down-regulate reactivity of endocrine thempy by participating in therapy resistance of PR negative breast cancer.
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Objective To investigate the influence of uremic serum on the monocyte chemoattractant protein 1 (MCP-1) expression of human umbilical vascular endothelial cells (HUVECs) in vitro and the effect of up-regulation of berne oxygenase 1 (HO-1) on the synthesis of MCP-1 of HUVECs in uremic milieu. Methods HUVECs were incubated to confluence and then preineubated with heroin and/or protoporphyrin zinc IX (ZnPP)for 6 hours.The cultures were subsequently incubated with M199 cell medium containing 10% serum of healthy people or with medium containing 10% serum of maintenance hemodialysis (MHD) patients.HO-1 protein and mRNA expression was detected by immunohistochemistry and semi-quantitative RT-PCR.MCP-1 mRNA expression was measured by semi-quantitative RT-PCR,and MCP-1 protein was quantified by ELISA. Results Up-regulated expression of MCP-1 mRNA and protein was detected in HUVECs incubated with medium containing 10% serum of MHD patients.The protein synthesis was 2.95 folds of the control.Heroin induced expression of HO-1 mRNA and protein,and concurrently inhibited the up-regulated MCP-1 expression induced by uremic serum.Such effects of heroin could be blocked by ZnPP. Conclusions Uremic serum induces the expression of MCP-1 in HUVECs.Up-regulated expresson of endothelial HO-1 induced by heroin inhibits the enhancement of MCP-1 synthesis.HO-1 may be beneficial to the alleviation of endothelial cell injury in uremic milieu.
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Objective To investigate the effect of cytochrome P_ 450 2A6 (CYP2A6) genetic polymorphisms on serum concentration of sodium valproate. Methods A total of 98 epileptic patients receiving sodium valproate after a period of more than 5 half-time were recruited. The genotypes of CYP2A6 of the patients were detected by nested-primer polymerase chain reaction (PCR) to examine the alleles CYP2A6*1 and CYP2A6*4. Fluorescence polarization immunoassay (FPIA) was used to measure the serum concentration of sodium valproate. Results Of the 98 cases, 73 (74.5%) were wild genotypes, 24 (24.5%) were CYP2A6*1/*4 genotypes and 1(1.0%) was CYP2A6*4/*4 genotype. According to the genotypes of CYP2A6 the patients were divided into two groups,one was group A (CYP2A6*1/*1) and the other was group B (CYP2A6*1/*4 or *4/*4). The mean value of the serum concentration of sodium valproate of the patients in group A(4.1393?0.2793) was higher than that in group B(3.3486?0.3919) with a statistical significance (P
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Objective To investigate the protective effect of heme oxygenase-1 for xeno-heart transplantation and the possible mechanism.Methods Cobalt protoporphyrin (CoPP), HO-1 inducer, and zinc protoporphyrin (ZnPP), HO-1 inhibitor, were used to intervene donors and recipients on the model of NIH-Wistar cardiac transplants respectively and simultaneously some of recipients were treated with immunesuppressor cyclosporine A (CsA), and control group and CsA treated group were set up respectively. The expression of HO-1 protein, HO-1 mRNA, caspase-3 and STAT-3 in transplanted heart tissue was detected by immunohistochemistry, RT-PCR and Western Blot. At the same time, HO-1 enzymatic activity was examined in heart tissue, and the cardiac cell apoptosis was examined by means of TUNEL. The differences among various factors were compared.Results The survival time of cardiac transplants in CoPP group was longer than that in control and ZnPP groups (P
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Objective To determine if hemin pretreatment has protective effects on the lungs in a rat model of septic shock.Methods Twenty healthy SD rats of both sexes aged 2.0-2.5 months, weighing 160-185 g were randomly divided into 2 groups (n=10): septic shock group (S) and hemin pretreatment group (H) . Septic shock was induced by intravenous LPS 10 mg?kg-1 in both groups. 24 h before induction of septic shock group H received hemin 100 mg?kg-1 intraperitoneally while group S received normal saline instead of hemin. Common carotid artery and external jugular vein were cannulated and hooked up to Hellige monitor. Evans blue 30 mg?kg-1 was given i.v. 5 min before LPS administration. The animals were sacrificed 6 h after intravenous LPS. The lungs were immediately removed for the determination of lung water content, SOD activity. Evans blue and malondialdehyde (MDA) contents, expression of HO-1 mRNA, HO-2 mRMA, and HO-1 protein, HO-2 protein levels. Results The lung water ratio (wet-dry lung weight/wet lung weight). Evans blue and MDA contents were significantly lower while SOD activity was significantly higher in group H than in group S (P
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Objective To investigate the expression of cyclooxygenase-2 in hepatocelluar carcinoma cell lines. Methods RT-PCR and immunocytochemistry were use d to investigate the expression of cyclooxygenase-2 in 6 hepatocellular carcino ma cell lines. Result COX-2 mRNA expression was detected in five of six cell lines, and all six cell lines w ere positive for COX-2 protein expression. Conclusion COX-2 is expressed in hepatocellular carcinoma cell li nes, providing basis for the chemoprevention of hepatocelluar carcinoma.
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Objective To explore the role of heme oxygenase-1 (HO-1) gene expression on murine experimental abdominal aortic aneurysm (AAA).Methods Wistar rats were divided into hemin (experimental group) and saline (control group) group randomly, and experimental AAA model was established by elastase perfusion. The specimen was obtained at postoperative day 7, and the dilatation rate was calculated. In situ hybridization was applied to detect the expression of HO-1 mRNA in aortic wall, while immunohistochemical staining was used to detect the protein expression of ICAM-1 and HO-1. Results In experimental group, the aorta dilation was inhibited and aneurysm was not observed. In experimental group, HO-1 mRNA and protein expression was strengthened (P