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This article reported a case of neonatal-onset autoinflammation with infantile enterocolitis (AIFEC) caused by NLRC4 gene mutation. The boy developed the disease in the neonatal period, presenting with recurrent fever, rash, hepatosplenomegaly and enterocolitis. Laboratory tests showed some indicators including ferritin and C-reactive protein were elevated. His condition was complicated by macrophage activation syndrome and anti-infective treatment was ineffective. High-throughput whole exome sequencing revealed a de novo heterozygous mutation of c.1021G>C (p.Val341Leu) in the NLRC4 gene and AIFEC was confirmed. AIFEC is a rare disease with no effective treatment at present, which can be developed in the neonatal period and diagnosed by whole exome sequencing.
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SUMMARY OBJECTIVE Analyze the over expression of neural precursor cell expressed developmentally down-regulated protein 9 (NEDD-9) deregulated associated with a poor prognosis in various carcinomas. Our objective was to investigate the relationship between the levels of NEDD-9, CA 15-3, and CEA and PET (SUVmax, MTV40, TLG40) with the clinical parameters of patients with breast cancer (BC). METHODS One hundred and eleven patients (82 BC patients who underwent 18F-FDG PET/CT and 29 healthy controls) were evaluated. SUVmax, MTV, and TLG of the primary tumor were compared with the molecular and histopathological subtypes. 18F-FDG, MTV, and TLG were evaluated based on the clinical data, i.e., nodal involvement, distant metastasis, ER and PR status, Ki-67, serum levels of NEDD-9, CA15-3, and CEA. We compared the NEDD-9 in the BC and healthy control groups. RESULTS The mean ± SD of SUVmax in the 82 patients was 13.0 ± 8.6. A statistically significant relationship (p = 0.022) was found between the molecular subtypes and 18F-FDG uptake. The relationship between 18F-FDG uptake and TLG measured in patients <50 years, ER-PR negativity, and HER2 positivity were statistically significant (p=0.015, 0.007, 0.046, and 0.001, respectively). MTV40, TLG40, and CA 15-3 in metastatic patients were statistically significant (p=0.004, 0.005, and 0.003, respectively). NEDD-9 in the BC group was significantly higher than in the healthy group (p=0.017). There was a positive correlation between SUVmax and Ki67 and CA 15-3; MTV40 and CEA; CA 15-3, CEA, SUVmax, and MTV40; a negative correlation was found between CEA, TLG40, and age. CONCLUSION The use of SUVmax, MTV40, and TLG40 parameters with NEDD-9 and tumor markers has been shown to provide a high diagnostic, predictive, and prognostic value for the management of BC. This is considered to be the basis of interventions focused on the treatment objectives related to NEDD-9.
RESUMO OBJETIVO Analisar a associação da superrexpressão das células NEDD-9 ao prognóstico negativo em vários tipos de carcinoma. Nosso objetivo foi investigar a relação entre os níveis de NEDD-9, CA 15-3 e CEA e PET (SUVmax, MTV40, TLG) e os parâmetros clínicos em pacientes com câncer de mama (CM). MÉTODOS Cento e onze pacientes (82 pacientes de CM submetidos a 18F-FDG PET/TC e 29 controles saudáveis) foram avaliados. SUVmax, MTV, e TLG do tumor primário foram comparados nos subtipos molecular e histopatológico. A captação de 18F-FDG, MTV, e TLG foi avaliada com base em dados clínicos (envolvimento nodal, metástase distante, status de ER e PR, Ki-67, níveis séricos de NEDD-9, CA15-3 e CEA). Foi comparada a NEDD-9 do grupo de CM e o controle saudável. RESULTADOS A média ± DP de SUVmax de 82 pacientes foi de 13,0 ± 8,6. Uma relação estatisticamente significativa (p=0,022) foi encontrada entre subtipos moleculares e captação de 18F-FDG. A relação entre captação de 18F-FDG e TLG medida em pacientes com idade <50 anos, ER-PR negativo e HER2 positivo foi estatisticamente significativa (p=0,015; 0,007; 0,046; e 0,001, respectivamente). MTV40, TLG40 e CA 15-3 em pacientes metastáticos foram estatisticamente significantes (p=0,004, 0,005 e 0,003, respectivamente). NEDD-9 no grupo BC foi significativamente maior do que no grupo saudável (p=0,017). Uma correlação positiva foi encontrada entre SUVmax e Ki67 e CA 15-3; MTV40 e CEA; CA 15-3, CEA, SUVmax e MTV40; uma correlação negativa foi encontrada entre CEA, TLG40 e idade. CONCLUSÃO O uso dos parâmetros SUVmax, MTV40 e TLG40 com NEDD-9 e marcadores tumorais demonstrou um alto valor diagnóstico, preditivo e prognóstico para o manejo do CM. Isso é considerado a base para intervenções focadas nos objetivos de tratamento relacionados às NEDD9.
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Humans , Breast Neoplasms/blood , Positron Emission Tomography Computed Tomography , Prognosis , Carcinoembryonic Antigen/blood , Tomography, X-Ray Computed , Retrospective Studies , Mucin-1/blood , Radiopharmaceuticals , Fluorodeoxyglucose F18 , Positron-Emission Tomography , Microtubule-Associated Proteins/bloodABSTRACT
Malignant tumors, whose occurrence and development are related to a variety of RNA transporter proteins, seriously affect human health and quality of life. Under normal circumstances, RNA transport proteins help RNA shuttle between nucleus and cytoplasm and their precise localization, effectively coupling the life activities in the nucleus and cytoplasm. During the process of tumorigenesis and progression, the expression and localization of some RNA transporters are abnormal or dysfunctional, which can change the subcellular localization, expression level, transport efficiency of downstream key RNA molecules, and the decay rate of cytoplasmic mRNA, and affect the proliferation, invasion and metastasis of tumors. This paper mainly reviews RNA transport proteins and their expression changes and regulation in tumors.
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Malignant tumors, whose occurrence and development are related to a variety of RNA transporter proteins, seriously affect human health and quality of life. Under normal circumstances, RNA transport proteins help RNA shuttle between nucleus and cytoplasm and their precise localization, effectively coupling the life activities in the nucleus and cytoplasm. During the process of tumorigenesis and progression, the expression and localization of some RNA transporters are abnormal or dysfunctional, which can change the subcellular localization, expression level, transport efficiency of downstream key RNA molecules, and the decay rate of cytoplasmic mRNA, and affect the proliferation, invasion and metastasis of tumors. This paper mainly reviews RNA transport proteins and their expression changes and regulation in tumors.
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Objective To investigate the effects of cisplatin on the expression of nucleotide － binding oligomerization domain－like receptor(NOD) 1 and 2 in human osteosarcoma SaOS－2 cell line,and to explore the mechanism of cisplatin in the treatment of human osteosarcoma. Methods CCK－8 assay,real－time quantitative reverse transcription polymerase chain reaction ( qRT － PCR ) and immumofluorescence methods were used to determine the growth survival rate and expression levels of NOD1 and NOD2 in osteosarcoma SaOS －2 cell line treated with cisplatin (0,5,10,20 μmol/L,named group S0,group S5,group S10,group S20,respectively) for 24, 48,72 hours. Results After treatment with cisplatin for 48 h or 72 h,the growth survival rates of SaOS－2 cells were significantly decreased in group S5 than those in group S0(65. 53% vs. 100. 00%;46. 43% vs. 100. 00%,χ2=8. 64,73. 97,all P＜0. 01). Moreover,after treatment with cisplatin for 24 h,48 h or 72 h,the growth survival rates of SaOS－2 cells were significantly decreased in group S10 or group S20 than those in group S0(80. 60% vs. 100. 00% , 42. 94 vs. 100. 00% ,27. 90% vs. 100. 00% ;62. 54% vs. 100. 00% ,33. 09% vs. 100. 00% ,22. 95% vs. 100. 00% , χ2=20. 99,79. 72,112. 50;45. 40,67. 56,125. 20,all P＜0. 01),and the growth survival rates were significantly lower in group S20 than those in group S5(62. 54% vs. 93. 78% ,33. 09% vs. 65. 53% ,22. 95% vs. 46. 43% ,χ2= 28. 47,21. 78,11. 71,all P ＜0. 01). The expression levels of NOD1 mRNA and NOD2 mRNA in group S5 were significantly increased at 48 h or 72 h than those at 24 h,and were higher than group S0 when treated with 5μmol/L cisplatin[(3. 64 ± 0. 44) vs. (4. 47 ± 1. 22) vs. (1. 79 ± 0. 44) vs (1. 00 ± 0. 00);(6. 88 ± 2. 79) vs. (6. 86 ± 2. 40) vs (2. 29 ± 0. 70) vs. (1. 00 ± 0. 00),F=29. 12,24. 11,all P＜0. 01]. And the expression levels of NOD1 protein had an increased tendency after 48 h or 72 h treatment with 5μmol/L cisplatin. Furthermore,the expression level of NOD1 mRNA was positively correlated with NOD2 mRNA(n=36,r=0. 92,P＜0. 01). Conclusion Cisplatin can elevate the function of osteosarcoma cell in a dose － and time － dependent manners, cisplatin may act as a efficient drug to cure osteosarcoma desease,which may be related to NOD1 and NOD2 signal pathway.
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Stimulator of interferon genes (STING) is a newly discovered adaptor protein in the innate immune system and plays an important role in innate immune response mediated by cytoplasmic DNA. Double-stranded DNA recognition receptors in cells are mediated by STING protein to produce type I interferon and other cytokines. Inadequate innate immune response and anti-hepatitis B virus (HBV) specific immune response are important causes of chronic HBV infection. This article introduces the discovery of STING and the latest research advances in its structure, briefly elaborates on the mechanism of activation of the STING signaling pathway, summarizes the research advances in the interaction between the STING signaling pathway and HBV, and points out the potential value of STING in clinical treatment.
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Objective To investigate the expression of Bcl-2 associated athanogene 3 (BAG3) in cervical cancer tissues and cells and its role in epithelial mesenchymal transition (EMT) of cervical cancer.Methods (1) Cervical cancer samples were collected from September 2015 to March 2017 in the Qilu Hospital of Shandong University and Shangdong Provincial Hospital.While,50 normal tissues were collected from August 2015 to March 2017 in the Dezhou Municiple Hospital,which were obtained from patients with uterine mnyoma underwent hysterectomy and patients with cervical biopsy.Reverse transcription (RT)-PCR and western blot were used to detect the expression of BAG3 mRNA and protein,and their clinical significances were analyzed.(2) The expression of BAG3 mRNA and protein was detected using RT-PCR and western blot method in HeLa and SiHa cell lines and normal cervical epithelial cells.The experiment was divided into two groups,BAG3 small interfering RNA transfected group (st-BAG3) and the control group transfected with small interfering RNA (siRNA).Cell counting kit 8 (CCK-8) analysis was used to detect cell proliferation of two groups.Wound-healing and transwell assay were used to detect the migration and invasion ability of HeLa and SiHa cells.The xenograft model of cervical cancer in nude mice was used to observe the effect of BAG3 on tumor xenografts and the tumor-related biomarkers were tested by western blot.Results (1) The expression levels of BAG3 mRNA and protein in cervical carcinoma tissues were 1.20±0.15 and 1.10±0.16,which were significantly higher than that in normal cervical tissue,0.23± 0.04 and 0.29 ± 0.03 (both P＜0.01).The results showed that the expression levels of BAG3 mRNA and protein were significantly correlated with cervical carcinoma staging and lymph node metastasis (P＜0.05).However,its expression was not conrelated with the patient's age,pathological grade,and diameter of tumor (all P＞0.05).(2) Compared with normal cervical epithelial cells,the expression of BAG3 mRNA and protein levels in HeLa and SiHa cells were significantly increased (P＜0.01),the expression levels of BAG3 mRNA and protein in HeLa and SiHa cells transfected with si-BAG3 were significantly lower than that in control group (all P＜0.01).After post-transfected 72 hours,A value of HeLa and SiHa with transfection were significantly lower than those in control group [(0.88±0.08) vs (1.22±0.13),(0.92±0.09) vs (1.35±0.12);both P＜0.01].After post-transfected 24 hours,the migration level of HeLa and SiHa cells with transfection were significantly lower than those in the control group [(20.1±2.1)％ vs (58.6±5.6)％,and (21.1±2.1)％ vs (61.7± 5.4)％;both P＜0.01].The transmembrane cell number in HeLa and SiHa cells with transfection were 76± 11 and 71±8,which were significantly less than those in control group (131± 12 and 129± 14;both P＜0.01).After the inoculation into nude mice,tumor formation time of HeLa and SiHa cells with transfection were (9.5±0.5) and (10.5 ± 1.3) days,respectively,which were significantly longer than those in control group [(4.5±0.5) and (5.2± 1.1) days;both P＜0.05].Compared with those in the control group,the expression level of Slug,N-cadherin and matrix metalloproteinase-2 (MMP-2) protein in HeLa and SiHa cells with transfected in tumor tissues were significantly decreased (all P＜0.01),while the expression level of E-cadberin protein was significantly increased (P＜0.01).Conclusion BAG3 could be involved in the proliferation,migration and invasion of cervical cancer cells by affecting cervical cancer EMT,and BAG3 may be an effective target for the treatment of cervical cancer.
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Objective To evaluate the changes in the expression of adaptor protein containing pleck-strin homobgy domain, phosphotyrosine-binding domain and a leucine zipper motif 1(APPL1)during renal fibrosis in a mouse model of renal ischemia-reperfusion(I∕R)injury. Methods Twenty-four male C57BL∕6 mice, aged 8 weeks, weighing 20-25 g, were divided into 2 groups(n=12 each)using a random number table: sham operation group(S group)and renal I∕R group.The model of renal I∕R injury was established by clipping the bilateral renal pedicles for 30 min followed by reperfusion in group I∕R.Six mice were selected at 2 days of reperfusion, and venous blood samples were collected for determination of serum concentrations of blood urea nitrogen and creatinine.The animals were then sacrificed, the renal specimens were obtained for microscopic examination of tubular necrosis with a light microscope, and the damage to the renal tubules was scored using a semi-quantitative method.Six mice were sacrificed at 14 days of reperfusion, and the renal specimens were obtained for assessment of the degree of renal fibrosis(using picric acid-sirius red staining) and for determination of the expression of collagen type 1, fibronectin and α-smooth muscle actin in renal tis-sues(by Western blot or immunofluorescence method). At 2 and 14 days of reperfusion, the expression of APPL1 in renal tissues was detected by Western blot and the expression of APPL1 mRNA in renal tissues by real-time polymerase chain reaction. Results Compared with group S, the serum concentrations of blood u-rea nitrogen and creatinine, scores of renal tubular damage and degree of renal fibrosis were significantly in-creased at 2 days of reperfusion, the expression of collagen type 1, fibronectin and α-smooth muscle actin in renal tissues was up-regulated at 14 days of reperfusion, and the expression of APPL1 protein and mRNA was up-regulated at 2 and 14 days of reperfusion in group I∕R(P<0.05). Conclusion Up-regulated expression of APPL1 may be involved in the process of renal fibrosis in a mouse model of renal I∕R injury.
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Objective To evaluate the relationship between placental expression of Gab1 and neonatal birth weight in mothers with gestational diabetes mellitus (GDM).Methods From the singleton and full-term cesarean delivered women in Shengjing Hospital Affiliated to China Medical University between October 2014 and May 2015,30 macrosomia babies with maternal GDM were selected as GDM macrosomia group,30 cases of GDM with normal neonatal birth weight as GDM normal group,30 cases without GDM but with macrosomia as normal macrosomia group,and 30 cases without GDM and with normal neonatal birth weight as the normal control group.Gab1 protein and mRNA expression in placentas were detected using immunohistochemistry,Western blot and real-time quantitative-polymerase chain reaction.Analysis of variance,LSD,Dunnett's T3,Chi-square test and Pearson's correlation analysis were used for statistical analysis.Results (1) Gab 1 protein location and positive expression rate:Gab 1 protein expression in human placenta tissue was located in the nucleus.The positive epression rate of Gab 1 protein in the GDM macrosomia group was higher than in the GDM normal group and normal macrosomia group [93％(28/30),73％(22/30) vs 73％(22/30)] and those in the normal macrosomia group and GDM normal group were higher than in the normal control group[47％(14/30)](x2=4.320,4.320,4.444 and 4.444,all P＜0.05).(2) The expression levels of Gabl protein and mRNA:The expression level of Gab1 protein in the GDM macrosomia group was higher than in the GDM normal group and normal macrosomia group (1.43 ± 0.58 vs 1.05 ± 0.67 and 0.95± 0.59),and that in the normal macrosomia group and GDM normal group were higher than in the normal control group (0.64±0.38) (LSD test,all P＜0.05).The expression levels of Gab1 mRNA showed the same trend as the expression levels of Gab1 protein in the four groups.(3) Gab 1 protein expression level was positively associated with neonatal birth weight (r=0.320,P=0.320).Conclusions The expression of Gab1 in placenta is involved in the regulation of birth weight in GDM mothers.
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Phosphoinositide-3 kinase/protein kinase-B/mammalian target of rapamycin (PI3K/Akt/mTOR) signaling pathway is involved in many important biological processes such as cell metabolism, growth, proliferation, and angiogenesis. And its high-level activation is closely related to the development and progression of many malignant tumors. In this paper, mTOR and PI3K/Akt/mTOR signaling pathway are introduced, and their action mechanisms in the development and progression of hepatocellular carcinoma, cholangiocellular carcinoma, cholangiocarcinoma, and gallbladder carcinoma are expounded, and then the role of mTOR inhibitors in the treatment of malignant hepatobiliary tumors is briefly described. It is thought that the PI3K/Akt/mTOR signaling pathway provides new therapeutic targets for malignant hepatobiliary tumors in advanced stage and the constant development of new mTOR inhibitors provides some new hope for the patients with malignant hepatobiliary tumors in advanced stage.
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BACKGROUND:The linkage and synergistic effect of adaptor proteins can effectively regulate signal transduction of T cel s, which can form a limit or amplification cascade to realize the complex immune function of T cel s. C-terminal Src kinase (Csk)-binding protein (Cbp) is an adaptor protein, which mainly exert the negative feedback regulation of Src kinase activity. This negative feedback effect depends on Y317 of Cbp, which may be involved in the SH2 domain of Csk. OBJECTIVE:To explore the effects of high expression of Cbp on ultrastructure and related biological function of Jurkat cel s. METHODS:The virus particles were constructed with expressing enhanced green fluorescent protein (EGFP) only and Cbp-EGFP fusion protein to transfect Jurkat cel s. There were untransfected group (Jurkat group), negative control group (transfected with expression of EGFP virus only), and Cbp group (transfected with Cbp-EGFP virus). RESULTS AND CONCLUSION:Confocal microscope showed that cel transfection efficiency was more than 95%and Cbp was located on the cel membrane. Optical microscope showed after transfection with Cbp-EGFP virus, more Jurkat cel s shrunk, with poor size uniformity. Apoptosis detection showed that after transfection with Cbp-EGFP virus, the number of apoptotic and necrotic cel s was greatly increased. Cbp mRNA expression was increased, Csk expression was decreased obviously and lymphocyte-specific protein tyrosine kinase expression was increased. So, in Jurkat cel s, the high expression of Cbp can decrease the uniformity of cel s and increase the necrosis cel s, thus inhibiting the signal transduction.
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BACKGROUND:Bone marrow mesenchymal stem cells can differentiate into osteoblasts under inducing condition that Zouguiwan and Youguiwan coordinate inducers, but the mechanism remains to be discussed. OBJECTIVE:To observe the effects of serum containing Zuoguiwan and Youguiwan on transforming growth factorβ1 and its signal transduction protein Smad2/3 message expression during the osteogenic differentiation of bone marrow mesenchymal stem cells. METHODS:A whole bone marrow adherence method was adopted to isolate and cultivate bone marrow mesenchymal stem cells from rats. The cellcultivation was processed in five groups:bone marrow mesenchymal stem cells were respectively cultured with blank serum, serum containing Zouguiwan, serum containing Youguiwan, positive serum containing progynova+inducer (dexamethasone, vitamin C, andβ-glycerophosphate), and inducer. Western blot was applied to detect the expression of type I col agen. The immunohistochemical assay was utilized to test transforming growth factorβ1 and Smad2/3 expression in the osteoblasts. RESULTS AND CONCLUSION:It was apparently more significant for serum containing Zuoguiwan and Youguiwan on type I col agen, transforming growth factorβ1 and Smad2/3 expression, compared with blank serum group and inducer group (P<0.05);moreover, serum containing Zuoguiwan was better than serum containing Youguiwan (P<0.05). Both of serum containing Zuoguiwan and Youguiwan are able to promote osteogenic differentiation of bone marrow mesenchymal stem cells. Moreover, Zuoguiwan is much more effective indicating that this method of traditional Chinese medicine about nourishing kidneys can be better to promote osteogenic induction of bone marrow mesenchymal stem cells.
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The primary causative factors of liver failure include direct damage and immune -mediated liver injury.Increasing evidence sug-gests that immune -mediated injury plays a pivotal role in the pathogenesis of liver failure.The new concepts concerning the mechanisms of immune -mediated liver injury in liver failure are reviewed with relevant basic and clinical studies in both humans and animals.The innate and adaptive immunity,particularly the interaction of various immune cells and molecules,as well as apoptosis -related molecules,are dis-cussed in detail.
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Phosphatidylinositol 3-kinase,Akt,and the mammalian target of rapamycin (PI3K/Akt/mTOR)signaling pathway is shown to play a key role in the tumorigenesis,proliferation,metastasis,apoptosis,and angiogenesis of hepatocellular carcinoma (HCC)by regulating gene expression.The components and functions of PI3K/Akt/mTOR signaling pathway are briefly described,and the research advances in the action mechanism of PI3K/Akt/mTOR signaling pathway in the progression of HCC and related inhibitors are reviewed.It is disclosed that blocking the PI3K/Akt/mTOR signaling pathway may become a new therapy for HCC.
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BACKGROUND:The proteome is a highlight technology in medical research fields lately, and has been reported to be applied in basic research fields related to liver transplantation. However, it has not been heard that the proteome has been used in research related to reduced-size liver transplantation. OBJECTIVE: To study expression of hepatic differential proteins related to signal transduction using proteomics after reduced-size liver transplantation in rats. METHODS:On the basis of successful establishment of rat models of reduced-size liver transplantation, transplanted liver tissues were obtained at 1, 3 and 7 days after transplantation. Postoperative liver tissue and normal donor, receptor liver tissues were subjected to solid pH gradient two-dimensional gel electrophoresis. Two-dimensional gel electrophoresis patterns were set up. Differentialy expressed protein spots were identified using tandem mass spectrometry analysis and database. RESULTS AND CONCLUSION:Seventy-two differential protein stains were found taking 10 times measure. Finaly, 32 proteins with clear functions were identified. Of them, four proteins participated in signal transduction, and they distributed at 3 and 7 days after liver transplantation, accounting for 6%. Results verified that on the basis of successful and stable establishment of rat models of reduced-size liver transplantation, proteomics technology was utilized to study differential proteins involving in signal transduction after reduced-size liver transplantation, and this study provides data for further deep investigation of regulating MicroRNA of these proteins.
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Objective To investigate the relationship between polycystic ovary syndrome (PCOS) susceptibility single nucleotide polymorphisms (SNP) and metabolic phenotypes in glucose and lipid metabolism and explore the pathophysiological mechanism of the susceptibility genes.Methods Three of PCOS susceptibility locus 2p16.3 (rs13405728 of LHCGR gene),2p21 (rs13429458,rs12478601 of THADA gene) and 9q33.3 (rs2479106,rs10818854 of DENNDIA gene) were selected and the metabolic phenotypes were compared between different genotypes of SNP in PCOS patients (using dominant model).Results Low-density lipoprotein cholesterol was significantly increased in CC genotype group than in TC + TT groups at rs12478601 of THADA gene [(2.5 ± 0.8),(2.4 ± 0.8) mmol/L; P =0.01].Serum insulin level of 2 hours after 75 g glucose intake was significantly higher in GG + AG groups than that of AA group at rs2479106 of DENND1A gene[(71 ±65),(64 ±50) mU/L;P =0.05],and the prevalence of type Ⅱ diabetes in first-degree relatives of patients were also increased [9.9％ (66/666),6.9％ (52/751) ; P ＜ 0.05].No association was found between metabolic phenotypes and genotypes of rs13429458,rs10818854,and rs13405728.Conclusions Genetic factors probably have effect on the metabolic characteristics of PCOS.THADA gene is related to lipid metabolism,while DENND1A gene may be involved in insulin metabolism in patients with PCOS.
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As interações microrganismo-hospedeiro se iniciam pela detecção de padrões moleculares associados a microrganismos (MAMPs) por receptores semelhantes à Toll (TLR) e por proteínas com domínio de ligação à nucleotídeos e oligomerização (Nod) na resposta imune inata. No entanto, como a cavidade bucal saudável é continuamente colonizada por microrganismos não patogênicos que também apresentam MAMPs, deve haver um mecanismo endógeno de regulação negativa da resposta do hospedeiro para evitar uma resposta exagerada e desnecessária com consequências negativas ao hospedeiro. Os mecanismos associados à distinção de microrganismos comensais e patogênicos na mucosa bucal são ainda pouco compreendidos. As proteínas Nod foram inicialmente descritas como 'TLRs intracelulares' capazes de reconhecer MAMPs no citosol; no entanto, estudos in vitro indicam que Nod têm papel relevante na regulação da expressão de RANKL e OPG induzidas por antígenos microbianos, bem como na modulação da atividade de vias de sinalização intracelular associadas à expressão de citocinas diretamente relacionadas à regulação do turnover do tecido ósseo. Devido à escassez de informações sobre o papel das proteínas Nod na modulação das interações microrganismo-hospedeiro na mucosa oral e com base nestas informações, nossa hipótese é que as proteínas Nod tem um papel relevante na modulação da reação inflamatória e suas consequências, incluindo a reabsorção do osso alveolar. Para testar esta hipótese, os objetivos específicos propostos foram: avaliar em camundongos knockout para Nod1, Nod2 ou Rip2, através de microtomografia computadorizada e avaliações histológicas descritivas, estereométricas e imunohistoquímicas (TRAP), o papel das proteínas Nod na inflamação e reabsorção óssea associadas à doença periodontal experimental induzida por bactérias inativadas por calor. A influência das proteínas Nod nas redes de citocinas e na sinalização intracelular associadas com a doença periodontal foi determinada in vitro em culturas primárias de macrófagos através de ensaios baseados nas plataformas ELISA e PCR em tempo real. Nossos resultados mostraram que, enquanto que nos camundongos Nod1 KO a reabsorção óssea alveolar e o número de osteoclastos aumentou significantemente, a deleção de Nod2 causou efeito contrário, diminuindo a reabsorção óssea. Os resultados in vitro foram correspondentes com os dados in vivo no caso de Nod2 (qPCR array, ELISA citocinas e vias de sinalização) e distintos no caso Nod1 (ELISA citocinas e vias de sinalização). Estes resultados demonstram que as proteínas Nod tem papel relevante na modulação das interações bactéria-hospedeiro associadas à doença periodontal induzida por bactérias inativadas por calor. Enquanto Nod1 parece exercer papel protetor na inflamação e reabsorção óssea, Nod2 atua como amplificador da resposta do hospedeiro
Recognition of pathogenic bacteria by the host is initially mediated by the innate immune response through detection of microbe-associated molecular pattern (MAMPs) by Toll-like receptors (TLR) and Nucleotide-oligomerization domain (Nod) proteins. Since the oral cavity, as well as other mucosal surfaces, is continuously colonized with non-pathogenic bacteria that also present MAMPs, there has to be an endogenous negative regulatory mechanism in place to prevent an overt host response with deleterious consequences. Specifically in the oral mucosa, it is not clear how the immune system is able to quickly distinguish between commensal and pathogenic bacteria and tailor the host response. Nod proteins were initially described as 'intracellular TLRs' that recognize MAMPs associated with bacteria invading the cytosol; however these proteins have been shown to modulate the activation of various signaling pathways involved in the expression of inflammatory genes, including MAPK and NF-κB in concert with TLR stimulation. There is paucity of information on the in vivo role of Nod proteins in the modulation of host-microbe interactions in the oral mucosa. Based on this information, our hypothesis is that Nod proteins play an important role in the modulation of the inflammatory reaction associated with periodontal diseases and its consequences, including alveolar bone resorption. To test this hypothesis, we propose the following specific aims: Assess the role of Nod proteins in the inflammation and bone resorption in experimentally-induced periodontal disease. Describe the influence of Nod proteins on the cytokine and signaling networks associated with periodontal disease
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In Vitro Techniques , Nod Signaling Adaptor Proteins , Periodontal Diseases , Bone ResorptionABSTRACT
Objective To study the relationship of Dll-1/Notch signal transduction pathway with the pathological characteristics of colorectal cancer and the effect on proliferation and apoptosis of colorectal cancer cells. Methods We assessed Notchl and Dll-1 protein levels in 63 cases of colorectal cancer and adjacent normal tissue by Western blotting.SW480 cells were treated with DAPT (γ-secretase inhibitor) at different treating times.MTT assay and flow cytometry were used to measure the proliferation and apoptosis of SW480 cells,seperately.The expression of the intracellular domain of Notch (NICD),Hes-1 and Bcl-2 were measured by Western blotting.Statistical methods were used including independent samples t test,paired sample t test and single factor analysis of variance. Results Notch1 and Dll-1 protein level increased in colorectal cancer tissues compared with adjacent normal mucosa,the mean values were 1.75-fold and 2.21-fold,respectively(t =2.554,P =0.012 and t =3.565,P =0.005).Also we found that the overexpression of Notch1 and Dl1-1 was related to the differentiation( t =2.463,P =0.017 and t =2.390,P=0.019),staging(t =2.675,P =0.007 and t =2.310,P =0.021) and lymph nodes metastasis(t =2.229,P =0.021 and t =2.210,P =0.023) of colorectal cancer.Treating SW480 cell with Notch pathway inhibitor (γ-secretase inhibitor,DAPT) resulted in growth inhibition,apoptosis induction and there was downregulation of NICD and Bcl-2 expression along with the treating time. Conclusions Overexpression of Notch1 and Dll-1 is related to the pathological characteristics of colorectal cancer.Blockade of Notch1 signal pathway may inhibit cell proliferation and induce cell apoptosis of colorectal cancer,as well as inhibit the expression of Bcl-2.
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Objective To evaluate the NLRP7 gene mutations and variants and their expression of genetic approach in hydatidiform mole patients with family history.Methods Six cases of mole patients with family members of mole history and 60 healthy women, taking blood, extracting DNA, the genetic mutation on NLRP7 screening and analysis, looking for mutations and corresponding amino acids, proteins control gene mutation found NLRP7 area.Results In 6 mole patients with family history:three patients were with sister's history of mole, and 2 of them familial recurrent hydatidiform mole(from family MoCh76 and family Ch77), there are 2 loci NLRP7 gene mutation.Screening patients from family MoCh76 for mutations in NLRP7 revealed in exon 3 and exon 5, amino acids [295G ＞ T] and [1970A ＞ T], proteins [Glu99X] and [Asp657Val], in a heterozygous.Screening patients from family Ch77 for mutations in NLRP7 revealed in exon4 and exon 7, amino acids [1294C ＞ T] and [2471 + 1G ＞ A], proteins [Arg432X] and [Leu825X], in a heterozygous.Screening patients from family 105 for mutations in NLRP7 revealed no NLRP7 gene mutation.There were mother's history of mole in three patients, and they were not familial recurrent hydatidiform mole.Screening patients from family MoCh73 for mutations in NLRP7revealed in exon 4, amino acids [1137G ＞ C], proteins [Lys379Asn], in a heterozygous.Screening patients from family 106 and family 110 for mutations in NLRP7 revealed no NLRP7 gene mutation.There were not found mutations and variations in 60 cases of ethnic matched control group.Conclusion NLRP7 mutations may be lead to familial recurrent hydatidiform mole.
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suitable tumor markers and new targets for gene therapy of cervical carcinoma.