RESUMO
Objective: Based on the genetic diagnosis and follow-up study on pediatric neurofibromatosis 1 (NF1) patients, interrogating the genotype-phenotype correlations of patients with NF1 mutations. Methods: 32 Patients from age of 2 months to 5 years old (17 male and 15 female) suspected for neurofibromatosis 1 were recruited during September 2016 to January 2018 in Shanghai Children's Medical Center retrospectively. Genetic diagnosis was applied to detect pathogenic variants. Long-term follow-up study were conducted to reveal progress of the disease and genotype-phenotype correlations. Results: 27 patients were detected with pathogenic NF1 variants, among them three were not reported. 3 patients inherited pathogenic variants from their NF1 diagnosed parents, all the other variants were de novo. Progressive development of phenotypes wasn't observed in most patients during the follow-up (14/27). Some patients were diagnosed with short stature, pulmonary artery stenosis and developmental delay during the follow-up(7/27). Short stature and pulmonary artery stenosis may be associated with missense mutation and severe truncation mutation of NF1 gene, respectively. Conclusions: Genetic diagnosis is required in young patients of NF1.Follow-up plan of pediatric patients should be adjusted based on genetic findings. Early follow-up of cardiovascular abnormalities should be noted in patients with missense mutation. Height development in patients with severe truncating variants are needed.
Assuntos
Neurofibromatose 1 , Criança , China , Feminino , Seguimentos , Humanos , Masculino , Neurofibromatose 1/diagnóstico , Neurofibromatose 1/genética , Neurofibromina 1 , Estudos RetrospectivosRESUMO
Objective: To explore the application and discovery of genotyping, gene sequencing, and gene expression analysis in the determination of ABO blood group subtypes and antigen expression abnormalities in hematological malignancies patients. Methods: From June 2019 to May 2020, three clinical cases were found with forward and reverse ABO typing discrepancy or atypical serologic agglutination pattern in the laboratory and blood transfusion department of Hebei Yanda Ludaopei Hospital were selected. Sequence-specific primer PCR (PCR-SSP) and Sanger sequencing of ABO gene coding regions were performed to determine the ABO genotypes, and whole transcriptome sequencing was used to analyze ABO and FUT1 gene expression levels. Results: A 12-year-old female acute lymphoblastic leukemia patient was determined as O.01.02 and BA.04 sub-genotype, corresponding to the serological B(A) subtype, and her ABO gene expression was normal (354.80). A 41-year-old female acute myeloid leukemia patient was determined as A1.02 and B.01 genotype, corresponding to the serological A(1)B phenotype, and her ABO gene expression was significantly reduced (45.70). A 42-year-old male with myelodysplastic syndrome and myelofibrosis was determined as A1.02 and A2.05 sub-genotype, corresponding to the serological A(1) and A(2) phenotype, respectively, and his ABO expression was negative. FUT1 expression was in the normal range in all three cases. The clinical blood product infusion strategy was formulated according to the genotype and the corresponding immunological subtype, and no significant transfusion-related adverse reactions occurred. Conclusion: Blood group sub-genotypes or aberrant gene expression can lead to ambiguities in serological blood group determination in hematological malignancies patients. ABO genotyping and gene expression analysis can help in this scenario and escort blood product infusion safety.
Assuntos
Tipagem e Reações Cruzadas Sanguíneas , Neoplasias Hematológicas , Sistema ABO de Grupos Sanguíneos/genética , Adulto , Alelos , Criança , Genótipo , Neoplasias Hematológicas/genética , Humanos , Masculino , FenótipoRESUMO
BACKGROUND: Diet and exercise during pregnancy have been used to prevent gestational diabetes mellitus (GDM) with some success. OBJECTIVE: To examine the effectiveness of lifestyle intervention on GDM prevention and to identify key effectiveness moderators to improve the prevention strategy. SEARCH STRATEGY: Pubmed, Scopus, Cochrane, and cross-references were searched. SELECTION CRITERIA: Randomised controlled trials (RCTs) evaluating lifestyle interventions during pregnancy for GDM prevention. DATA COLLECTION AND ANALYSIS: Two independent reviewers extracted data. A random-effects model was used to analyse the relative risk (RR) and 95% confidence interval (95% CI). Meta-regressions and subgroup analyses were used to investigate important moderators of effectiveness. MAIN RESULTS: Forty-seven RCTs involving 15 745 participants showed that diet and exercise during pregnancy were preventive of GDM (RR 0.77, 95% CI 0.69-0.87). Four key aspects were identified to improve the preventive effect: targeting the high-risk population; an early initiation of the intervention; the correct intensity and frequency of exercise; and gestational weight gain management. Although 24 RCTs targeted women who were overweight or obese, body mass index (BMI) failed to predict the effectiveness of an intervention. Instead, interventions are most effective in high-incidence populations rather than simply in women who are overweight or obese. Furthermore, exercise of moderate intensity for 50-60 minutes twice a week could lead to an approximately 24% reduction in GDM. CONCLUSION: The best strategy to prevent GDM is to target the high-risk population predicted by risk evaluation models and to control the gestational weight gain of women through intensified diet and exercise modifications early in their pregnancy. TWEETABLE ABSTRACT: Four key effectiveness moderators of lifestyle interventions for GDM prevention.
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Diabetes Gestacional/prevenção & controle , Dietoterapia , Terapia por Exercício , Obesidade/terapia , Complicações na Gravidez/terapia , Feminino , Humanos , Sobrepeso/terapia , Gravidez , Análise de RegressãoRESUMO
Objective: To establish the method of detecting the concentrations of diphenylamine in air of workplace with high performance liquid chromatographic (HPLC) . Methods: According to standards of methods for determining the chemical substances in workplace air, diphenylamine in the air was collected by glass fiber filter treated with sulfuric acid, then dissolved by methanol and determined by high performance liquid chromatography with UV-detector. Results: There was a linear relationship within the range of 0-30.0 µg/ml, and regression equation was y=8425.6x-150.7, correlation coefficient was 0.999 9, the detection limit was 0.045 µg/ml. The lowest detected concentration was 0.030 mg/m(3) (sampling volume 15 L) . The within-run precision was 2.41 â -3.02%, the between-run precision was 3.11%-4.45%. The desorption efficiencies was 97.8â and the sampling efficiencies were 100%. The samples in glass fiber filter could be stored for 7 d at room temperature. Conclusion: The present method could meet with the requirements of Guide for establishing occupational health standards-Part 4 Determination methods of air chemicals in workplace and be feasible for determination of diphenylamine in workplace air.
Assuntos
Poluentes Ocupacionais do Ar/análise , Cromatografia Líquida de Alta Pressão/métodos , Difenilamina/análise , Local de Trabalho , Ar , Monitoramento Ambiental/métodos , Humanos , Limite de DetecçãoRESUMO
OBJECTIVE: To analyze the role of acetylsalicylic acid (ASA) in immunomodulation of mesenchymal stem cells derived from gingiva (GMSCs), and to explore the role of ASA in enhancing the immumomodulation of GMSCs and the capacity of GMSCs to treat immune disorders and the underlying mechanism. METHODS: Flow cytometry analysis were used to analyze the role of ASA in the expression of stem cells surface markers CD146, CD105, CD90, CD34 and CD45 in GMSCs,and the GMSCs proliferation was analyzed by 5-bromo-2-deoxyuridine (BrdU) staining and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The GMSCs and T cells co-culture system was established to analyze the role of ASA in immunomodulation of GMSCs by measuring T cell apoptosis using flow cytometry analysis and inflammatory cytokines using enzyme linked immunosorbent assay (ELISA). Further more, the dextran sulfate sodium (DSS) induced colitis mouse model was established and the mouse body weight, disease activity score, histological index and pathological change of colons were analyzed after GMSC infusion. RESULTS: The proliferation of GMSCs and the expressions of CD105, CD146 in GMSCs were increased after ASA treatment. In the GMSCs and T cells co-culture system, GMSCs induced T cells apoptosis and inhibited interferon γ (IFN-γ) and tumor necrosis factor α (TNF-α) secretion by T cells, which were enhanced by ASA treatment. In vivo, GMSCs infusion could ameliorate DSS-induced colitis, including inhibited DSS-induced mouse body weight loss, decreased disease activity score and histological index, and decreased inflammation cells infiltration in colons, as shown by hematoxylin-eosin (HE) staining. Moreover, the therapeutic effects of GMSC infusion on DSS-induced colitis could be enhanced by ASA treatment. Mechanically, ASA treatment increased FasL expression of Fas/FasL death pathway in GMSCs to induce T cells apoptosis. CONCLUSION: ASA enhanced immunomodulation of GMSCs and increased the capacity of GMSCs to ameliorate DSS-induced colitis in mice.
Assuntos
Anti-Inflamatórios não Esteroides , Aspirina , Colite , Gengiva , Imunomodulação , Células-Tronco Mesenquimais , Animais , Anti-Inflamatórios não Esteroides/farmacologia , Apoptose , Aspirina/farmacologia , Colite/imunologia , Sulfato de Dextrana , Modelos Animais de Doenças , Gengiva/citologia , Gengiva/imunologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/imunologia , Camundongos , Camundongos Endogâmicos C57BLRESUMO
Objective: To evaluate the effect of endoplasmic reticulum stress in trophocytes, in patients with intrahepatic cholestasis of pregnancy (ICP). Methods: Sixty-one pregnant women who were hospitalized in Women's Hospital, School of Medicine, Zhejiang University from January to December 2015 were recruited. Thirty-one women who were diagnosed as ICP were defined as the ICP group and 30 healthy pregnant women were defined as the control group. The localization and expression intensity of glucose regulated protein 78 (GRP-78) in placental tissues were detected by immunohistochemistry technique. Electronic microscope was used to observe ultra-microstructure change of the endoplasmic reticulum in trophocytes and cell line Swan71. Reverse transcription (RT)-PCR and western blot were used to investigate the expression of GRP-78 mRNA and protein in Swan 71 cell. Results: (1) GRP-78 protein was mainly expressed in the cytoplasm of cytotrophoblasts and syncytiotrophoblasts. The protein expression of GRP-78 in placentas of the ICP group (13.2±2.4) was significantly higher than that in the control group (7.8±1.3, P<0.01). (2) The volume of endoplasmie reticulum did not increase and the microvilli developed well, with no swelling and no expansion of endoplasmic reticulum in the control group.In the ICP group, microvilli injury, endoplasmic reticulum edema were found; the volume of endoplasmic reticulum increased, with dilation, vacuolation and significant degranulation. After treated with 100 µmol/L cholyglycine for 24 hours, universal dilatation of the endoplasmic reticulum were seen in the Swan71 cells. (3) In Swan71 cells, cholylglycine displayed a concentration-dependent up-regulation on the expression of GRP-78. The expressions of GRP-78 mRNA in 0, 25, 50, 100 µmol/L cholylglycine experimental group were 1.01±0.17, 2.17±0.16, 5.47±0.36, 5.65±0.82, respectively. The expression of GRP-78 protein in 0, 25, 50, 100 µmol/L cholylglycine experimental group were 1.01±0.04, 1.17±0.15, 1.33±0.13, 1.73±0.13, respectively. The expression of GRP-78 mRNA and protein in 100 and 50 µmol/L cholylglycine experimental group were significantly higher than 0 µmol/L (all P<0.01). Conclusion: The obvious expansion of endoplasmic reticulum and the increased expression of GRP-78 in trophocytes indicated that endoplasmic reticulum stress of trophocytes may be involved in the pathogenesis of ICP.
Assuntos
Colestase Intra-Hepática/patologia , Estresse do Retículo Endoplasmático , Retículo Endoplasmático/metabolismo , Proteínas de Choque Térmico/genética , Placenta/metabolismo , Complicações na Gravidez/patologia , Animais , Western Blotting , Estudos de Casos e Controles , Chaperona BiP do Retículo Endoplasmático , Feminino , Ácido Glicocólico , Proteínas de Choque Térmico/metabolismo , Humanos , Gravidez , Terceiro Trimestre da Gravidez , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Trofoblastos , Regulação para CimaRESUMO
OBJECTIVE: To explore possible mechanism of ERBB2 gene expression silencing mediating mitogen-activated protein kinase 1/mitogen-activated protein kinase 3 (MAPK1/MAPK3) signaling pathway on proliferation, migration, and invasion of ovarian cancer cells. PATIENTS AND METHODS: A total of 240 cancer specimens were collected in patients with epithelial ovarian cancer intraoperatively in our hospital from January 2015 to January 2018. Expressions of ERBB2, MAPK1, and MAPK3 in tissues were detected by immunohistochemistry. Following the culture of ovarian cancer cell lines, target cell line with high expression of ERBB2 was screened by qRT-PCR. Cell grouping was performed with four groups after transfection, including Blank group, negative control (NC) group, ERBB2 shRNA group, and ERBB2 overexpression group (shorted as ERBB2 group). The expression levels of ERBB2, MAPK1, MAPK3, vascular endothelial growth factor (VEGF), metalloproteases-2 (MMP-2), and tissue inhibitor of metalloproteases-2 (TIMP-2) were detected by qRT-PCR in different transfection groups, followed by the detection of protein expressions with Western blot. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to test the proliferation activity of each group after transfection, while transwell assay and scratch test explored cell invasion and migration in each group, respectively. RESULTS: Immunohistochemistry showed that the positive rates of ERBB2, MAPK1, and MAPK3 in ovarian cancer tissues were significantly increased than those in adjacent normal epithelial tissues. In the cell experiment, ERBB2 gene was highly expressed in SKOV3 ovarian cancer cell line. There was no significant difference in each index between Blank group and NC group (p > 0.05). Compared with Blank group and NC group, the expression levels of ERBB2, MAPK1, MAPK3, VEGF, and MMP-2 in ERBB2 shRNA group decreased significantly, TIMP-2 increased markedly, and proliferation, invasion, and migration abilities of cells decreased markedly after transfection, showing statistically significant differences (All p < 0.05). By contrast, the expression levels of ERBB2, MAPK1, MAPK3, VEGF, and MMP-2 increased remarkably in ERBB2 group, while TIMP-2 decreased significantly, and cell proliferation, invasion, and migration ability increased evidently after transfection, with statistically significant differences (All p < 0.05). CONCLUSIONS: Silencing ERBB2 gene expression may inhibit the activation of MAPK1/MAPK3 signaling pathway and thus suppress the proliferation, invasion, and migration of ovarian cancer cells. Overexpression of ERBB2 gene can reverse those trends, which in turn support the role of ERBB2 gene expression silencing in molecular targeted therapy of ovarian cancer.
Assuntos
Movimento Celular , Inativação Gênica , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Neoplasias Ovarianas/metabolismo , Neoplasias Ovarianas/patologia , Receptor ErbB-2/genética , Transdução de Sinais , Células Cultivadas , Feminino , Humanos , Proteína Quinase 1 Ativada por Mitógeno/genética , Proteína Quinase 3 Ativada por Mitógeno/genética , Invasividade Neoplásica , Receptor ErbB-2/metabolismo , Transdução de Sinais/genéticaRESUMO
OBJECTIVE: This study aims to clarify the potential function of ATAD2 (ATPase family, AAA domain containing 2) in regulating proliferative and apoptotic abilities of colorectal carcinoma (CRC). PATIENTS AND METHODS: ATAD2 levels in CRC specimens and cell lines were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Overall survival in CRC patients with high or low level of ATAD2 was assessed by Kaplan-Meier method. The correlation between ATAD2 level and clinical characteristics of CRC patients was analyzed by χ2 test. Univariable and multivariable Cox regression models were generated to illustrate potential risk factors for the overall survival of CRC. After knockdown of ATAD2 in SW620 cells, relative levels of Cyclin D1, ppRb, pRb, E2F1, Cyclin E and cleaved Caspase 3 were detected by Western blot. Regulatory effects of ATAD2 on viability, clonality, cell cycle distribution, and apoptosis in SW620 and HCT15 cells were examined by a series of functional experiments. RESULTS: Upregulated ATAD2 in CRC was correlated to tumor size, tumor node metastasis (TNM) staging, and histological classification of CRC. High level of ATAD2 predicted poor prognosis in CRC patients. Cox regression test suggested that ATAD2 level, tumor size, TNM staging and histological classification were independent factors influencing overall survival in CRC. Knockdown of ATAD2 reduced viability and clonality in SW620 and HCT15 cells. In addition, cell cycle was arrested in G1 phase and apoptosis was stimulated in CRC cells with ATAD2 knockdown. In SW620 cells transfected with ATAD2 shRNA, protein levels of Cyclin D1, ppRb, E2F1 and Cyclin E were downregulated, and cleaved Caspase 3 was upregulated. CONCLUSIONS: ATAD2 is upregulated in CRC tissues and correlated to poor prognosis of CRC patients. It exerts an anti-proliferation role in CRC by arresting cell cycle in G1/S phase and triggering apoptosis via the Rb-E2F1 signaling.
Assuntos
ATPases Associadas a Diversas Atividades Celulares/genética , Proliferação de Células , Neoplasias Colorretais/genética , Neoplasias Colorretais/metabolismo , Proteínas de Ligação a DNA/genética , Fator de Transcrição E2F1/metabolismo , Transdução de Sinais , ATPases Associadas a Diversas Atividades Celulares/metabolismo , Apoptose , Células Cultivadas , Neoplasias Colorretais/patologia , Proteínas de Ligação a DNA/metabolismo , Fator de Transcrição E2F1/genética , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Transdução de Sinais/genéticaRESUMO
OBJECTIVE: To study the role of HGF (stem cell growth factor) in renal interstitial fibrosis and to explore its underlying mechanism. MATERIALS AND METHODS: A unilateral ureteral obstruction (UUO) mouse model was first constructed, and kidney samples of mice were then collected. Fibrosis-related indicators in UUO mice kidney were detected by Western blot. The mRNA and protein levels of HGF in UUO mice were detected by quantitative Real-time-polymerase chain reaction (qRT-PCR) and Western blot, respectively. The HGF overexpression mouse model was established by using UUO mice. For in vitro experiments, fibrosis-related indicators and the expression of HGF were detected in transforming growth factor-ß1 (TGF-ß1)-induced NRK-52E cells. Finally, a p-SMAD3 knockdown mouse model was established to confirm whether p-SMAD3 was involved in HGF-regulated renal interstitial fibrosis. RESULTS: The expression levels of HGF and α-SMA (α-smooth muscle actin) were both significantly increased in UUO mice, while E-cadherin expression was significantly decreased, which were consistent with results of in vitro experiments. Overexpression of HGF remarkably decreased the protein and mRNA levels of α-SMA in fibrotic NRK-52E cells. After overexpression of HGF in UUO mice, α-SMA was remarkably downregulated, whereas E-cadherin was significantly upregulated. Further, results also demonstrated that HGF was upregulated and α-SMA was downregulated after p-SMAD3 knockdown in UUO mice. CONCLUSIONS: HGF is highly expressed during renal interstitial fibrosis, which may suppress renal interstitial fibrosis by inhibiting the TGF-ß1/SMAD signaling pathway.
Assuntos
Fator de Crescimento de Hepatócito/uso terapêutico , Nefrite Intersticial/tratamento farmacológico , Transdução de Sinais/efeitos dos fármacos , Proteína Smad3/antagonistas & inibidores , Fator de Crescimento Transformador beta1/antagonistas & inibidores , Animais , Caderinas/biossíntese , Linhagem Celular , Fibrose/tratamento farmacológico , Fibrose/patologia , Técnicas de Silenciamento de Genes , Masculino , Camundongos , Nefrite Intersticial/patologia , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Proteína Smad3/biossíntese , Proteína Smad3/genética , Obstrução Ureteral/complicaçõesRESUMO
Objective: To investigate the association between human papillomavirus (HPV) and lung cancer. Methods: We examined a series of 83 lung cancer patients with HPV DNA in both lung tumor specimens and adjacent normal specimens from Fujian province. Twenty-one of the most clinically relevant HPV types from the highly conserved L1 region of the viral genome were analyzed, using the PCR amplification and were followed by reverse hybridization with specific probes. Chi-square test of paired design was used to test the difference of HPV positive rates between lung cancer specimens and adjacent normal specimens. Chi-square test and Fisher's exact test were used to analyze the differences of HPV positive rate of tumor specimens on factors as gender, age, histological subtype, clinical stage, smoking status and alcohol consumption. Results: HPV was detected in 7 of the 83 tumor specimens and in 6 of the paired normal lung tissues. There was no significant correlation between HPV and lung cancer (P>0.999). Neither demographic characteristics nor clinical features were found with significant differences on HPV in lung cancer tissues (P>0.05). Conclusion: Our data showed that HPV was not significantly associated with the risk of lung cancer in Fujian province.
Assuntos
Neoplasias Pulmonares , Infecções por Papillomavirus , Humanos , Papillomaviridae , Reação em Cadeia da Polimerase , FumarRESUMO
OBJECTIVES: Glutamine is an important fuel for intestinal mucosal epithelial cells, and it promotes intestinal mucosal cell differentiation and proliferation. Most liver transplantation (LT) patients suffer from intestinal barrier dysfunction. Whether enteral glutamine supplementation has beneficial effects on intestinal barrier function following LT is not known. We investigated the effect of glutamine (Gln) supplementation on NF-κB and on the intestinal barrier in rats after an allogenic LT with concomitant immunosuppressive therapy. MATERIALS AND METHODS: Inbred Sprague-Dawley rats (n=40) receiving allogenic LT were randomly divided into Gln and control groups (n=20, each). Gln group rats were administered Gln (0.4 g/kg·day) by gastric infusion for 6 days, while control rats received saline. Ten rats from each group were sampled for basal parameters on the 3rd day, prior to LT. The remaining 10 from each group were sampled after receiving LT. Twenty inbred Sprague-Dawley rats were selected as donors. The 20 recipients underwent orthotopic LT after 3 days of treatment and were given immunosuppressive therapy for 6 days post-operation. They were euthanized for sample collection on the 7th day. NF-κB protein in the intestinal mucosa, portal plasma Gln, endotoxin and TNF-α levels, ileocecal sIgA content, bacterial translocation and mucosal ultrastructure were assessed. RESULTS: On the postoperative day 6, the Gln group had increased plasma Gln and ileocecal sIgA (secretory IgA). Gln group also showed improvement in mucosal microvilli structure and had reduced levels of intestinal mucosal NF-κB, portal endotoxin and TNF-α and decreased bacterial translocation as compared to the control group. CONCLUSIONS: Parenteral supplementation of glutamine ameliorated mucosal injury during allogenic LT, and improved intestinal barrier function. These findings suggest that glutamine supplementation may be an effective therapy to ensure successful recovery from liver transplantation.
Assuntos
Glutamina/farmacologia , Mucosa Intestinal/efeitos dos fármacos , Intestinos/efeitos dos fármacos , Transplante de Fígado/métodos , Animais , Suplementos Nutricionais , Glutamina/sangue , Imunossupressores/farmacologia , Mucosa Intestinal/fisiologia , Intestinos/fisiologia , Masculino , Distribuição Aleatória , Ratos , Ratos Sprague-DawleyRESUMO
AIM: L-arginine (L-Arg) is an aminoacid that has immunomodulating and antitumor effects. It is possible that antitumor effects of L-Arg are due to induction of apoptosis in tumor cells. The present study assessed antiproliferating and proapoptotic effects of L-Arg in human gastric cancer cell line SGC-7901. METHODS: Cell proliferation was quantified by MTT assay. Apoptosis was assessed using flow cytometry and FITC-Annexin-V/propidium iodide staining. Expression and activation of proteins pertinent to apoptosis (Bcl-2, surviving, p53, and XIAP) were studied using PCR, Western blot, and activity assays. RESULTS: L-Arg significantly inhibited growth of SCG-7901 gastric cancer cells and downregulated expression of antiapoptotic gene Bcl-2 and survivin. By contrast, expression of p53 was upregulated by L-Arg. CONCLUSION: Regulation of apoptosis by L-Arg via downregulation of antiapoptotic proteins Bcl-2 and surviving, and upregulation of proapoptotic protein p53 may represent the mechanism behind antitumor effects of L-Arg.
Assuntos
Apoptose , Arginina/química , Neoplasias Gástricas/tratamento farmacológico , Neoplasias Gástricas/patologia , Linhagem Celular Tumoral , Proliferação de Células , Regulação Neoplásica da Expressão Gênica , Humanos , Proteína Supressora de Tumor p53/metabolismoRESUMO
Aim: L-arginine (LArg) is an amino acid that has immunomodulating and anti-tumor effects. It is possible that anti-tumor effects of L-Arg are due to induction of apoptosis in tumor cells. The present study assessed anti-proliferating and pro-apoptotic effects of L-Arg in human gastric cancer cell line SGC7901. Methods: Cell proliferation was quantified by MTT assay. Apoptosis was assessed using flow cytometry and FITCAnnexinV/ propidium iodide staining. Expression and activation of proteins pertinent to apoptosis (Bcl2, surviving, p53, and XIAP) were studied using PCR, Western blot, and activity assays. Results: L-Arg significantly inhibited growth of SCG-7901 gastric cancer cells and down-regulated expression of anti-apoptotic gene Bcl-2 and survivin. By contrast, expression of p53 was upregulated by L-Arg. Conclusion: Regulation of apoptosis by L-Arg via down-regulation of anti-apoptotic proteins Bcl-2 and surviving, and up-regulation of pro-apoptotic protein p53 may represent the mechanism behind antitumor effects of L-Arg.
RESUMO
We present a biosensor design based on capturing the two-dimensional (2D) phase image of surface plasmon resonance (SPR). This 2D SPR imaging technique may enable parallel label-free detection of multiple analytes and is compatible with the microarray chip platform. This system uses our previously reported differential phase measurement approach, in which 2D phase maps obtained from the signal (P) and reference (S) polarizations are compared pixel by pixel. This technique greatly improves detection resolution as the subtraction step can eliminate measurement fluctuations caused by external disturbances as they essentially appear in both channels. Unlike conventional angular SPR systems, in which illumination from a range of angles must be used, phase measurement requires illumination from only one angle, thus making it well suited for 2D measurement. Also, phase-stepping introduced from a moving mirror provides the necessary modulation for accurate detection of the phase. In light of the rapidly increasing need for fast real-time detection, quantification, and identification of a range of proteins for various biomedical applications, our 2D SPR phase imaging technique should hold a promising future in the medical device market.
Assuntos
Biopolímeros/análise , Técnicas Biossensoriais/instrumentação , Análise de Sequência com Séries de Oligonucleotídeos/instrumentação , Óptica e Fotônica/instrumentação , Processamento de Sinais Assistido por Computador/instrumentação , Ressonância de Plasmônio de Superfície/instrumentação , Técnicas Biossensoriais/métodos , Desenho de Equipamento , Análise de Falha de Equipamento , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Ressonância de Plasmônio de Superfície/métodosRESUMO
Creeping bentgrass (Agrostis palustris Huds.) is a cool season grass widely used on putting greens in golf courses. Transformation of creeping bentgrass has been conducted using microprojectile bombardment and protoplast electroporation. The objective of our study is to develop an alternative and more efficient approach in transforming the grass using Agrobacterium (strain EHA 101). This technique was effective in transforming 40-day old calli derived from mature seeds cultured on MS medium supplemented with 2,4-D, kinetin, and sucrose. Dozens of transgenic plants have been produced from two independent transformed calli. Presence of functional green fluorescence protein (GFP) was detected in leaves, stems, and roots of transgenic seedlings. Four putative transgenic plants and two control plants were randomly chosen and analyzed by Southern blot analysis. Bands corresponding to the GFP gene were clearly shown in transgenic plants. These results indicated that Agrobacterium transformation can successfully be applied to creeping bentgrass.
Assuntos
Adenina/análogos & derivados , Genes Reporter , Proteínas Luminescentes/genética , Plantas Geneticamente Modificadas , Poaceae/genética , Rhizobium/genética , Transformação Genética , Adenina/farmacologia , Southern Blotting , Eletroporação/métodos , Proteínas de Fluorescência Verde , Cinetina , Modelos Genéticos , Regiões Promotoras Genéticas , Sacarose/farmacologia , Fatores de TempoRESUMO
From September 1990 to February 1991, 15 cases of myocardial revascularization with the right gastroepiploic artery (RGEA) combined with arterial or venous grafts were performed at VGH-Taipei. All were males, ranging in age from 49 to 64 years (mean age 61.2 +/- 4.6 years). The mean number of distal anastmoses including vein grafts was 3.6 +/- 0.7 and the mean number of graft was 3.1 +/- 0.3 per patient. The mean aortic clamp time was 123.6 +/- 24.0 minutes and the mean cardiopulmonary bypass time was 176 +/- 35 minutes. There was one mortality (6.7%). The other 14 patients are alive without angina. Studied within 3 postoperative months, graft early patency was 100% (6/6) in GEA graft. GEA graft should be a third available arterial conduit for coronary artery bypass.
Assuntos
Ponte de Artéria Coronária/métodos , Idoso , Ponte de Artéria Coronária/efeitos adversos , Humanos , Masculino , Pessoa de Meia-Idade , Estômago/irrigação sanguínea , Grau de Desobstrução VascularRESUMO
In addition to beta-sitosterol and alpha-amyrin detected in all the investigated species, the extract of the aerial parts of Artemisia giraldii var. giraldii gave stigmasterol, daucosterol, sesamine, luteolin, eupafolin, hispidulin, eupatilin, belamcanidin, pinitol, artemin, ridentin, and a new antifungal monoterpene (named santolinylol) while that of the aerial parts of A. mongolica afforded sesamine, eupafolin, eupatilin, matricarin, and a new germacranolide (3-oxo-11 alpha H-germacra-1(10)E,4Z-dien-12,6 alpha-olide), and that of the aerial parts of A. vestita yielded stigmasterol, daucosterol, umbelliferone, scopolin, scoparone, and isoscopoletin-O-glucoside. Pinitol, first reisolated from Artemisia genus, was shown to inhibit the growth of the human pathogenic fungi Candida albicans, Aspergillus flavus, A. niger, Geotrichun candidum, Trichophyton rubrum, and Epidermophyton floccosum. Umbelliferone was also active against Candida tropicalis, A. flavus, G. candidum, T. rubrum, and E. floccosum. The flavones hispidulin and belamcanidin were almost equally inhibitory to the growth of A. flavus, G. candidum, T. rubrum, and E. floccosum, and santolinylol to C. albicans, A. flavus, A. niger, G. candidum, T. rubrum, and E. floccosum. In addition, ridentin was active against the growth of the plant pathogenic fungus Cladosporium cucumerinum.