RESUMO

PURPOSE: CD44v6 plays a controversial role in tumor progression and patient outcome in colorectal cancer by plenty of conflicting reports. The purpose of this study was to profile the intratumoral heterogeneity of CD44v6 in rectal cancer and investigate its role in lymph node metastasis. METHODS: Sixty patients were included in this study. Immunohistochemistry for CD44v6 was performed in normal mucosa, primary tumor, and lymph node metastasis with whole tissue sections. The staining intensity in tumor center and invasive front was separately measured. Sampling bias was evaluated by quantitative real-time PCR with 15 pairs of frozen tissues from different sites of the primary tumor. RESULTS: CD44v6 expression increased from normal mucosa to primary tumor to lymph node metastasis. Multiple intratumoral staining patterns was observed in primary tumor, and CD44v6 expression in invasive front was significantly higher than that in tumor center. In addition, mRNA expression levels differed across different geographical regions of the tumor. No association between CD44v6 expression and lymph node metastasis was revealed. CONCLUSIONS: Substantial intratumoral heterogeneity of CD44v6 exists in rectal cancer that impacts the outcome of individual studies. CD44v6 expression should be assessed in a more precise way with a specified staining pattern and in a designated location.

Assuntos

Adenocarcinoma/secundário , Biomarcadores Tumorais/metabolismo , Receptores de Hialuronatos/metabolismo , Mucosa/metabolismo , Neoplasias Retais/patologia , Adenocarcinoma/genética , Adenocarcinoma/metabolismo , Biomarcadores Tumorais/genética , Feminino , Seguimentos , Humanos , Receptores de Hialuronatos/genética , Técnicas Imunoenzimáticas , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Mucosa/patologia , Gradação de Tumores , Invasividade Neoplásica , Prognóstico , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Neoplasias Retais/genética , Neoplasias Retais/metabolismo , Estudos Retrospectivos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Taxa de Sobrevida

RESUMO

Helicobacter pylori employs unique methods to colonize the stomach, which induces chronic inflammation. It is also able to avoid eradication by macrophages and other immune cells. Leukocyte cell-derived chemotaxin 2 (LECT2), a multi-functional cytokine involved in many pathological conditions, has recently been shown to activate macrophages via the CD209a receptor. Therefore, we aimed to investigate the effects of LECT2 on H. pylori-infected macrophages. Macrophages were treated with recombinant LECT2, and both their ability to kill H. pylori and produce nitric oxide were analyzed. Western blot was performed to determine nuclear translocation and protein phosphorylation of p65, a subunit of nuclear factor (NF)-κB. Transfection experiments were performed to analyze the signaling pathway of LECT2 in macrophages. We found that treatment with LECT2 enhanced H. pylori killing and nitric oxide production in macrophages. In addition, DNA-binding activity and nuclear translocation of p65 were up-regulated by LECT2 treatment. Furthermore, we found that NF-κB activation by LECT2 was mediated by Raf-1 in macrophages, and Raf-1 phosphorylation was specifically altered in response to LECT2. Moreover, LECT2 induced Ser28 phosphorylation in the intracellular domain of CD209a. CD209a Ser28 phosphorylation was required for LECT2-induced Raf-1 and NF-κB activation in RAW264.7 macrophages. Our study showed that the effects of LECT2 on H. pylori killing and nitric oxide production were dependent on CD209a phosphorylation, Raf-1, and NF-κB activation. Together, these results demonstrate for the first time that exposure to LECT2 can modulate specific intracellular mechanisms downstream of CD209a to enhance H. pylori killing and nitric oxide production in macrophages.

Assuntos

Infecções por Helicobacter/genética , Peptídeos e Proteínas de Sinalização Intercelular/genética , Proteínas Recombinantes/genética , Fator de Transcrição RelA/biossíntese , Animais , Moléculas de Adesão Celular/genética , Moléculas de Adesão Celular/imunologia , Infecções por Helicobacter/imunologia , Infecções por Helicobacter/microbiologia , Helicobacter pylori/imunologia , Helicobacter pylori/patogenicidade , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/farmacologia , Lectinas Tipo C/genética , Lectinas Tipo C/imunologia , Macrófagos/imunologia , Camundongos , Óxido Nítrico/biossíntese , Óxido Nítrico/genética , Fosforilação , Células RAW 264.7 , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/imunologia , Proteínas Recombinantes/farmacologia , Transdução de Sinais/genética , Estômago/imunologia , Estômago/microbiologia , Fator de Transcrição RelA/genética , Ativação Transcricional/genética , Transfecção

RESUMO

Genetic relationships of 17 Rhododendron cultivars, China, were assessed using inter-simple sequence repeat (ISSR) and amplified fragment length polymorphism (AFLP) markers. A total of 133 bands were obtained using nine selected ISSR primers, 129 (96.99%) of which were polymorphic; 267 bands were amplified by four AFLP primer pairs, 251 (94.01%) of which exhibited polymorphism. Based on these polymorphic products, a cluster analysis revealed similarities between the results of the ISSR and AFLP. All of the cultivars were clustered into two major branches; one branch contained the same four cultivars, and the other cultivars were separated into different groups in the other branch. The cluster results showed that the genetic relationships of the 17 cultivars were partly related to their morphological characteristics, particularly the flowering phase. Therefore, the results of this study support the classification of Rhododendron cultivars according to flowering phase. In addition, the cluster results can be used to select suitable parents for breeding.

Assuntos

Flores/genética , Rhododendron/genética , Flores/crescimento & desenvolvimento , Estudos de Associação Genética , Repetições de Microssatélites , Filogenia , Polimorfismo de Fragmento de Restrição , Rhododendron/crescimento & desenvolvimento

RESUMO

Fluoride, which is often added to toothpaste or mouthwash in order to protect teeth from decay, may be a novel therapeutic approach for acceleration of periodontal regeneration. Therefore, we investigated the effects of fluoride on proliferation and mineralization in human periodontal ligament cells in vitro. The periodontal ligament cells were stimulated with various concentrations of NaF added into osteogenic inductive medium. Immunohistochemistry of cell identification, cell proliferation, alkaline phosphatase (ALP) activity assay, Alizarin red S staining and quantitative real-time-polymerase chain reaction (RT-PCR) were performed. Moderate concentrations of NaF (50-500 µmol/L) had pro-proliferation effects, while 500 µmol/L had the best effects. ALP activity and calcium content were significantly enhanced by 10 µmol/L NaF with osteogenic inductive medium. Quantitative RT-PCR data varied in genes as a result of different NaF concentrations and treatment periods. We conclude that moderate concentrations of NaF can stimulate proliferation and mineralization in periodontal ligament cells. These in vitro findings may provide a novel therapeutic approach for acceleration of periodontal regeneration by addition of suitable concentrations of NaF into the medication for periodontitis treatment, i.e., into periodontal packs and tissue patches.

Assuntos

Proliferação de Células/efeitos dos fármacos , Ligamento Periodontal/efeitos dos fármacos , Fluoreto de Sódio/farmacologia , Adolescente , Adulto , Fosfatase Alcalina/efeitos dos fármacos , Fosfatase Alcalina/metabolismo , Cálcio/metabolismo , Células Cultivadas/efeitos dos fármacos , Criança , Humanos , Ligamento Periodontal/citologia , Reação em Cadeia da Polimerase em Tempo Real/métodos , Adulto Jovem

RESUMO

This study reports a case of a gonadotropin-releasing hormone agonist trigger in a young female with myelodysplastic syndrome (MDS) who underwent fertility preservation using random-start controlled ovarian stimulation. This method involves the stimulation of the ovary regardless of a patient's menstrual-cycle phase. A review of the related literature is also provided. A 17-year-old patient was diagnosed with MDS and required initiation of peripheral blood stem cell transplantation within a maximum of 3 weeks and was in the luteal phase of the menstrual cycle when the possibility of attempting preservation of fertility was presented to her. She opted for a random-start controlled ovarian stimulation with gonadotropins. With successful hemorrhagic prophylaxis, 17 oocytes were retrieved including 10 mature and 7 immature oocytes. Of the immature oocytes, 3 were successfully matured in vitro and a vitrification protocol was used to freeze the 13 mature oocytes.

Assuntos

Preservação da Fertilidade/métodos , Síndromes Mielodisplásicas/fisiopatologia , Indução da Ovulação/métodos , Adolescente , Criopreservação/métodos , Feminino , Humanos , Ciclo Menstrual/fisiologia , Recuperação de Oócitos/métodos , Oócitos/fisiologia , Reprodutibilidade dos Testes , Resultado do Tratamento

RESUMO

We investigated the risk factors for pulmonary hypertension (PH) in patients receiving maintenance peritoneal dialysis (MPD). A group of 180 end-stage renal disease patients (124 men and 56 women; mean age: 56.43±8.36) were enrolled in our study, which was conducted between January 2009 and June 2014. All of the patients received MPD treatment in the Dialysis Center of the Second Affiliated Hospital of Soochow University. Clinical data, laboratory indices, and echocardiographic data from these patients were collected, and follow-ups were scheduled bi-monthly. The incidence and relevant risk factors of PH were analyzed. The differences in measurement data were compared by t-test and enumeration data were compared with the χ2 test. Among the 180 patients receiving MPD, 60 were diagnosed with PH. The remaining 120 were regarded as the non-PH group. Significant differences were observed in the clinical data, laboratory indices, and echocardiographic data between the PH and non-PH patients (all P<0.05). Furthermore, hypertensive nephropathy patients on MPD showed a significantly higher incidence of PH compared with non-hypertensive nephropathy patients (P<0.05). Logistic regression analysis showed that the proportion of internal arteriovenous fistula, C-reactive protein levels, and ejection fraction were the highest risk factors for PH in patients receiving MPD. Our study shows that there is a high incidence of PH in patients receiving MPD and hypertensive nephropathy patients have an increased susceptibility to PH.

Assuntos

Humanos , Masculino , Feminino , Pessoa de Meia-Idade , Idoso , Fístula Arteriovenosa/complicações , Hipertensão Pulmonar/etiologia , Diálise Peritoneal/efeitos adversos , Proteína C-Reativa/análise , China/epidemiologia , Hipertensão Pulmonar/epidemiologia , Incidência , Falência Renal Crônica/complicações , Falência Renal Crônica/terapia , Peptídeo Natriurético Encefálico/sangue , Fósforo/sangue , Estudos Prospectivos , Fatores de Risco

RESUMO

This study evaluated the effect of muscle satellite cells (MSCs) overexpressing myogenin (MyoG) on denervated muscle atrophy. Rat MSCs were isolated and transfected with the MyoG-EGFP plasmid vector GV143. MyoG-transfected MSCs (MTMs) were transplanted into rat gastrocnemius muscles at 1 week after surgical denervation. Controls included injections of untransfected MSCs or the vehicle only. Muscles were harvested and analyzed at 2, 4, and 24 weeks post-transplantation. Immunofluorescence confirmed MyoG overexpression in MTMs. The muscle wet weight ratio was significantly reduced at 2 weeks after MTM injection (67.17±6.79) compared with muscles injected with MSCs (58.83±5.31) or the vehicle (53.00±7.67; t=2.37, P=0.04 and t=3.39, P=0.007, respectively). The muscle fiber cross-sectional area was also larger at 2 weeks after MTM injection (2.63×10³±0.39×10³) compared with MSC injection (1.99×10³±0.58×10³) or the vehicle only (1.57×10³±0.47×10³; t=2.24, P=0.049 and t=4.22, P=0.002, respectively). At 4 and 24 weeks post-injection, the muscle mass and fiber cross-sectional area were similar across all three experimental groups. Immunohistochemistry showed that the MTM group had larger MyoG-positive fibers. The MTM group (3.18±1.13) also had higher expression of MyoG mRNA than other groups (1.41±0.65 and 1.03±0.19) at 2 weeks after injection (t=2.72, P=0.04). Transplanted MTMs delayed short-term atrophy of denervated muscles. This approach can be optimized as a novel stand-alone therapy or as a bridge to surgical re-innervation of damaged muscles.

Assuntos

Transplante de Células , Denervação Muscular/reabilitação , Músculo Esquelético/inervação , Atrofia Muscular/reabilitação , Miogenina/metabolismo , Células Satélites de Músculo Esquelético/transplante , Animais , Imunofluorescência , Expressão Gênica , Masculino , Atrofia Muscular/etiologia , Miogenina/genética , Tamanho do Órgão/genética , Plasmídeos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo Real , Células Satélites de Músculo Esquelético/citologia , Células Satélites de Músculo Esquelético/metabolismo , Neuropatia Ciática/reabilitação , Transfecção

RESUMO

We investigated the risk factors for pulmonary hypertension (PH) in patients receiving maintenance peritoneal dialysis (MPD). A group of 180 end-stage renal disease patients (124 men and 56 women; mean age: 56.43±8.36) were enrolled in our study, which was conducted between January 2009 and June 2014. All of the patients received MPD treatment in the Dialysis Center of the Second Affiliated Hospital of Soochow University. Clinical data, laboratory indices, and echocardiographic data from these patients were collected, and follow-ups were scheduled bi-monthly. The incidence and relevant risk factors of PH were analyzed. The differences in measurement data were compared by t-test and enumeration data were compared with the χ2 test. Among the 180 patients receiving MPD, 60 were diagnosed with PH. The remaining 120 were regarded as the non-PH group. Significant differences were observed in the clinical data, laboratory indices, and echocardiographic data between the PH and non-PH patients (all P<0.05). Furthermore, hypertensive nephropathy patients on MPD showed a significantly higher incidence of PH compared with non-hypertensive nephropathy patients (P<0.05). Logistic regression analysis showed that the proportion of internal arteriovenous fistula, C-reactive protein levels, and ejection fraction were the highest risk factors for PH in patients receiving MPD. Our study shows that there is a high incidence of PH in patients receiving MPD and hypertensive nephropathy patients have an increased susceptibility to PH.

Assuntos

Fístula Arteriovenosa/complicações , Hipertensão Pulmonar/etiologia , Diálise Peritoneal/efeitos adversos , Idoso , Proteína C-Reativa/análise , China/epidemiologia , Feminino , Humanos , Hipertensão Pulmonar/epidemiologia , Incidência , Falência Renal Crônica/complicações , Falência Renal Crônica/terapia , Masculino , Pessoa de Meia-Idade , Peptídeo Natriurético Encefálico/sangue , Fósforo/sangue , Estudos Prospectivos , Fatores de Risco

RESUMO

OBJECTIVE: This study aims to observe the clinical efficacy of low-dose rituximab in patients with cryoglobulinaemia secondary to connective tissue diseases. METHODS: Rituximab (100 mg) was infused in patients once a week for four weeks, combined with prednisone (20 mg) once a day, and reduced regularly. Treatment effect was observed regularly. RESULTS: Joint pain, fever, rash and fatigue symptoms in patients eased. The serology (globulin, erythrocyte sedimentation rate, C-reactive protein, lactate dehydrogenase and others) parameters returned to normal. CONCLUSION: Low-dose rituximab therapy for cryoglobulinaemia secondary to connective tissue diseases was safe and effective, and can be used as a treatment option in this condition.

RESUMO

This study evaluated the effect of muscle satellite cells (MSCs) overexpressing myogenin (MyoG) on denervated muscle atrophy. Rat MSCs were isolated and transfected with the MyoG-EGFP plasmid vector GV143. MyoG-transfected MSCs (MTMs) were transplanted into rat gastrocnemius muscles at 1 week after surgical denervation. Controls included injections of untransfected MSCs or the vehicle only. Muscles were harvested and analyzed at 2, 4, and 24 weeks post-transplantation. Immunofluorescence confirmed MyoG overexpression in MTMs. The muscle wet weight ratio was significantly reduced at 2 weeks after MTM injection (67.17±6.79) compared with muscles injected with MSCs (58.83±5.31) or the vehicle (53.00±7.67; t=2.37, P=0.04 and t=3.39, P=0.007, respectively). The muscle fiber cross-sectional area was also larger at 2 weeks after MTM injection (2.63×103±0.39×103) compared with MSC injection (1.99×103±0.58×103) or the vehicle only (1.57×103±0.47×103; t=2.24, P=0.049 and t=4.22, P=0.002, respectively). At 4 and 24 weeks post-injection, the muscle mass and fiber cross-sectional area were similar across all three experimental groups. Immunohistochemistry showed that the MTM group had larger MyoG-positive fibers. The MTM group (3.18±1.13) also had higher expression of MyoG mRNA than other groups (1.41±0.65 and 1.03±0.19) at 2 weeks after injection (t=2.72, P=0.04). Transplanted MTMs delayed short-term atrophy of denervated muscles. This approach can be optimized as a novel stand-alone therapy or as a bridge to surgical re-innervation of damaged muscles.

Assuntos

Animais , Masculino , Atrofia Muscular/reabilitação , Miogenina/metabolismo , Transplante de Células , Músculo Esquelético/inervação , Células Satélites de Músculo Esquelético/transplante , Denervação Muscular/reabilitação , Tamanho do Órgão/genética , Plasmídeos , Atrofia Muscular/etiologia , Transfecção , Expressão Gênica , Imunofluorescência , Ratos Sprague-Dawley , Miogenina/genética , Células Satélites de Músculo Esquelético/citologia , Reação em Cadeia da Polimerase em Tempo Real

RESUMO

Fluoride, which is often added to toothpaste or mouthwash in order to protect teeth from decay, may be a novel therapeutic approach for acceleration of periodontal regeneration. Therefore, we investigated the effects of fluoride on proliferation and mineralization in human periodontal ligament cells in vitro. The periodontal ligament cells were stimulated with various concentrations of NaF added into osteogenic inductive medium. Immunohistochemistry of cell identification, cell proliferation, alkaline phosphatase (ALP) activity assay, Alizarin red S staining and quantitative real-time-polymerase chain reaction (RT-PCR) were performed. Moderate concentrations of NaF (50-500 μmol/L) had pro-proliferation effects, while 500 μmol/L had the best effects. ALP activity and calcium content were significantly enhanced by 10 μmol/L NaF with osteogenic inductive medium. Quantitative RT-PCR data varied in genes as a result of different NaF concentrations and treatment periods. We conclude that moderate concentrations of NaF can stimulate proliferation and mineralization in periodontal ligament cells. These in vitro findings may provide a novel therapeutic approach for acceleration of periodontal regeneration by addition of suitable concentrations of NaF into the medication for periodontitis treatment, i.e., into periodontal packs and tissue patches.

Assuntos

Humanos , Criança , Adolescente , Adulto , Adulto Jovem , Proliferação de Células/efeitos dos fármacos , Ligamento Periodontal/efeitos dos fármacos , Fluoreto de Sódio/farmacologia , Fosfatase Alcalina/efeitos dos fármacos , Fosfatase Alcalina/metabolismo , Cálcio/metabolismo , Células Cultivadas/efeitos dos fármacos , Ligamento Periodontal/citologia , Reação em Cadeia da Polimerase em Tempo Real/métodos

RESUMO

This study reports a case of a gonadotropin-releasing hormone agonist trigger in a young female with myelodysplastic syndrome (MDS) who underwent fertility preservation using random-start controlled ovarian stimulation. This method involves the stimulation of the ovary regardless of a patient's menstrual-cycle phase. A review of the related literature is also provided. A 17-year-old patient was diagnosed with MDS and required initiation of peripheral blood stem cell transplantation within a maximum of 3 weeks and was in the luteal phase of the menstrual cycle when the possibility of attempting preservation of fertility was presented to her. She opted for a random-start controlled ovarian stimulation with gonadotropins. With successful hemorrhagic prophylaxis, 17 oocytes were retrieved including 10 mature and 7 immature oocytes. Of the immature oocytes, 3 were successfully matured in vitro and a vitrification protocol was used to freeze the 13 mature oocytes.

Assuntos

Humanos , Feminino , Adolescente , Preservação da Fertilidade/métodos , Síndromes Mielodisplásicas/fisiopatologia , Indução da Ovulação/métodos , Criopreservação/métodos , Ciclo Menstrual/fisiologia , Recuperação de Oócitos/métodos , Oócitos/fisiologia , Reprodutibilidade dos Testes , Resultado do Tratamento

RESUMO

Internal transcribed spacer 1 (ITS1) sequences from wild-type Exopalaemon carinicauda (N = 124) from the East China Sea were amplified and sequenced. Sequences were polymorphic and ranged from 388 to 583 bp in length. The average content of GC in sequences was significantly higher than that of AT. Altogether, 604 mutant sites with 123 haplotypes were detected; 46.7% were polymorphic sites. The genetic diversity index of population Y was highest, and the lowest was population X. Eight microsatellite sequences were detected; the most-repeated sequences were (GA)n, (AG)n, (GT)n, (TG)n, (TC)n, and (CT)n. Analysis of molecular variance revealed that genetic differentiation among the four populations were very weak, or modest. A molecular evolutionary tree was constructed using the neighbor-joining method and MEGA 6.0, and the phyletic evolutionary relationships among several Palaemonidae species examined. The phylogenetic tree showed that individuals of the same species, as well as the species of the same genus, clustered together, consistent with morphological classifications.

Assuntos

DNA Intergênico/genética , Palaemonidae/genética , Penaeidae/classificação , Penaeidae/genética , Animais , China , Genética Populacional , Filogenia , Alinhamento de Sequência

RESUMO

The role of microRNAs (miRNAs) in the regulation of mammalian reproduction has been demonstrated previously. However, only a few studies have assessed the role of miRNAs in the reproduction processes of sheep. The elucidation of miRNA expression profiles in the ovaries of different sheep breeds representing fecundity extremes will be useful in understanding the roles of miRNAs in sheep reproduction. In this study, two small RNA libraries were constructed from ovary tissue taken from Qira black sheep and Hetian sheep during the estrous period and then sequenced using the Solexa sequencing method. We obtained 9,565,212 and 9,563,426 high-quality reads from Qira black sheep and Hetian sheep, respectively. In total, 531 miRNAs, including 98 putative miRNAs, were identified. Among the conserved miRNAs, 125 known miRNAs were significantly differentially expressed in the Qira black sheep and Hetian sheep libraries, with 24 upregulated and 101 downregulated in the Hetian sheep compared to the Qira black sheep. Four differentially expressed miRNAs were analyzed using real-time quantitative PCR to validate the reliability of the Solexa sequencing results. These results provide a foundation for future research on the regulation of miRNAs in sheep fertility and enrich the sheep miRNA databases.

Assuntos

MicroRNAs/biossíntese , MicroRNAs/genética , Ovário/metabolismo , Ovinos/genética , Animais , Feminino , Sequenciamento de Nucleotídeos em Larga Escala/veterinária , MicroRNAs/análise , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Análise de Sequência de RNA/veterinária , Transcriptoma

RESUMO

Several previous studies have demonstrated that elevated levels of fibroblast growth factor-23 (FGF-23) may be involved in atherosclerosis and contribute to the high mortality rate of peritoneal dialysis (PD) patients. The aim of this study was to determine the precise role of FGF-23 in the pathogenesis of atherosclerosis in PD patients. Between April 2009 and January 2012, 62 PD patients and 25 control subjects were included in the study. An enzyme-linked immunosorbent assay was conducted to test for plasma FGF-23 levels. Carotid artery intima-media thickness (CIMT), left ventricular mass index (LVMI), and myocardial performance index (MPI) were determined by ultrasonography. Plasma Ca(2+), P(3+), calcium-phosphorus product, parathyroid hormone, N-terminal pro-brain natriuretic peptide, and cardiac troponin I were also detected. Plasma FGF-23 levels in PD patients were significantly higher than those in control subjects. PD patients with CIMT > 1.0 mm showed the highest levels of FGF-23. Plasma P(3+), calcium-phosphorous product, plasma parathyroid hormone, CIMT, LVMI, and MPI levels were positively associated with plasma FGF-23 levels. Multiple-stepwise regression analyses revealed that plasma P(3+), plasma parathyroid hormone, CIMT, LVMI, and MPI levels were strongly associated with plasma FGF-23 levels. However, no correlations were observed in plasma N-terminal pro-brain natriuretic hormone and cardiac troponin I levels. Plasma FGF- 23 levels may play an important role in the initiation and progression of atherosclerosis. Thus, detecting and defining plasma FGF-23 levels may be a promising biomarker for the early detection of atherosclerosis in PD patients.

Assuntos

Aterosclerose/genética , Biomarcadores/sangue , Fatores de Crescimento de Fibroblastos/sangue , Diálise Peritoneal , Adulto , Idoso , Aterosclerose/sangue , Aterosclerose/patologia , Feminino , Fator de Crescimento de Fibroblastos 23 , Humanos , Masculino , Pessoa de Meia-Idade , Peptídeo Natriurético Encefálico/sangue , Hormônio Paratireóideo/sangue , Fatores de Risco , Troponina I/sangue

RESUMO

MicroRNAs (miRNAs) are small RNA molecules that modulate gene expression implicated in cancer, which play crucial roles in diverse biological processes, such as development, differentiation, apoptosis, and proliferation. The aim of this study was to investigate whether miR-30c mediated the resistance of breast cancer cells to the chemotherapeutic agent doxorubicin (ADR) by targeting tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta (YWHAZ). miR-30c was downregulated in the doxorubicin-resistant human breast cancer cell lines MCF-7/ADR and MDA-MB-231/ADR compared with their parental MCF-7 and MDA-MB-231 cell lines, respectively. Furthermore, we observed that transfection of an miR-30c mimic significantly suppressed the ability of MCF-7/ADR to resist doxorubicin. Moreover, the anti-apoptotic gene YWHAZ was confirmed as a target of miR-30c by luciferase reporter assay, and further studies indicated that the mechanism for miR-30c on the sensitivity of breast cancer cells involved YWHAZ and its downstream p38 mitogen-activated protein kinase (p38MAPK) pathway. Together, our findings provided evidence that miR-30c was one of the important miRNAs in doxorubicin resistance by regulating YWHAZ in the breast cancer cell line MCF-7/ADR.

Assuntos

Antibióticos Antineoplásicos/farmacologia , Doxorrubicina/farmacologia , Resistencia a Medicamentos Antineoplásicos , MicroRNAs/fisiologia , Animais , Resistencia a Medicamentos Antineoplásicos/genética , Ativação Enzimática/efeitos dos fármacos , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Concentração Inibidora 50 , Células MCF-7 , Camundongos , MicroRNAs/genética , Triptofano Hidroxilase/efeitos dos fármacos , Tirosina 3-Mono-Oxigenase/efeitos dos fármacos

RESUMO

In response to salinity or drought stress, many plants accumulate glycine betaine, which is a regulator of osmosis. In plants, the last step in betaine synthesis is catalyzed by betaine aldehyde dehydrogenase (BADH), a nuclear-encoded chloroplastic enzyme. Based on the conserved oligo amino acid residues of the published BADH genes from other higher plant species, a cDNA sequence, designated CtBADH, was isolated from safflower (Carthamus tinctorius L.) using a polymerase chain reaction approach. The clones were 1.7 kb on average, and contained an open reading frame predicting a polypeptide of 503 amino acids with 84.5% identity to that of Helianthus annuus. The deduced amino acid sequence showed a decapeptide, Val-Thr-Leu-Geu-Leu-Gly-Gly-Lys-Ser-Pro and Cys, which is essential for proper functioning of BADH. Phylogenetic analysis indicated that CtBADH grouped with other dicotyledonous plant BADH genes, and subgrouped in the composite family. Prediction of secondary structure and subcellular localization suggested that the protein encoded by CtBADH contains 33 coils, 15 alpha helixes, and 21 beta strands, and most likely targets the chloroplast or mitochondria.

Assuntos

Betaína-Aldeído Desidrogenase/genética , Carthamus tinctorius/enzimologia , Genes de Plantas , Proteínas de Plantas/genética , Sequência de Aminoácidos , Sequência de Bases , Betaína-Aldeído Desidrogenase/química , Betaína-Aldeído Desidrogenase/metabolismo , Carthamus tinctorius/química , Carthamus tinctorius/genética , Clonagem Molecular , Dados de Sequência Molecular , Filogenia , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Estrutura Terciária de Proteína , Transporte Proteico

RESUMO

MicroRNAs (miRNAs) are small RNA molecules that modulate gene expression implicated in cancer, which play crucial roles in diverse biological processes, such as development, differentiation, apoptosis, and proliferation. The aim of this study was to investigate whether miR-30c mediated the resistance of breast cancer cells to the chemotherapeutic agent doxorubicin (ADR) by targeting tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta (YWHAZ). miR-30c was downregulated in the doxorubicin-resistant human breast cancer cell lines MCF-7/ADR and MDA-MB-231/ADR compared with their parental MCF-7 and MDA-MB-231 cell lines, respectively. Furthermore, we observed that transfection of an miR-30c mimic significantly suppressed the ability of MCF-7/ADR to resist doxorubicin. Moreover, the anti-apoptotic gene YWHAZ was confirmed as a target of miR-30c by luciferase reporter assay, and further studies indicated that the mechanism for miR-30c on the sensitivity of breast cancer cells involved YWHAZ and its downstream p38 mitogen-activated protein kinase (p38MAPK) pathway. Together, our findings provided evidence that miR-30c was one of the important miRNAs in doxorubicin resistance by regulating YWHAZ in the breast cancer cell line MCF-7/ADR.

Assuntos

Animais , Feminino , Humanos , Camundongos , Antibióticos Antineoplásicos/farmacologia , Resistencia a Medicamentos Antineoplásicos , Doxorrubicina/farmacologia , MicroRNAs/fisiologia , Resistencia a Medicamentos Antineoplásicos/genética , Ativação Enzimática/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , MicroRNAs/genética , Triptofano Hidroxilase/efeitos dos fármacos , /efeitos dos fármacos

RESUMO

PURPOSE: Emerging evidences indicate that dysregulated microRNAs are implicated in cancer tumorigenesis and progression. MicroRNA-9 (miR-9) has various expression patterns in diverse human cancers. However, its clinical significance in human non-small cell lung cancer has not yet been elucidated. In the present study, we detected the expression of miR-9 in non-small cell lung cancer and adjacent noncancerous tissues and explored its relationships with clinicopathological characteristics and prognosis. METHODS: Expression levels of miR-9 in 116 pairs of non-small cell lung cancer and adjacent normal tissues were detected by real-time quantitative RT-PCR assay. To determine its prognostic value, overall survival (OS) and progression-free survival (PFS) were evaluated using the Kaplan-Meier method. Univariate and multivariate analysis were performed using the Cox proportional hazard analysis. RESULTS: MiR-9 expression in non-small cell lung cancer tissues was significantly higher than that in adjacent normal tissues (p = 0.001), and its up-regulation was significantly correlated to advanced tumor-node-metastasis (TNM) stage (p < 0.001), tumor size (p = 0.013), and lymph node metastasis (p = 0.001). Furthermore, Kaplan-Meier analysis demonstrated that high miR-9 expression clearly predicted poorer PFS (p < 0.001) and OS (p < 0.001). In the multivariate analysis, increased miR-9 expression was an independent prognostic factor for both PFS (p = 0.002) and OS (p = 0.013). CONCLUSIONS: MiR-9 was up-regulated in non-small cell lung cancer tissues and correlated with adverse clinical features and unfavorable survival, indicating that miR-9 might be involved in non-small lung cancer progression and could serve as a promising biomarker for further risk stratification in the treatment of this cancer.

Assuntos

Biomarcadores Tumorais/genética , Carcinoma Pulmonar de Células não Pequenas/genética , Regulação Neoplásica da Expressão Gênica , Neoplasias Pulmonares/genética , MicroRNAs/genética , Regulação para Cima/genética , Idoso , Carcinoma Pulmonar de Células não Pequenas/mortalidade , Feminino , Humanos , Neoplasias Pulmonares/mortalidade , Masculino , Pessoa de Meia-Idade , Taxa de Sobrevida

RESUMO

OBJECTIVE: Changes in soluble intercellular adhesion molecule-1 (sICAM-1) and E-selectin levels as well as leukocyte count were examined in this study to explore the relationship between leukopenia and ICAMs in Graves' disease (GD). METHODS: Fasting blood samples were obtained from 37 GD patients with normal leukocytes and 32 GD patients with leukopenia. Enzyme-linked immunosorbent assay (ELISA) was performed to determine serum sICAM-1 and E-selectin levels for comparison. The same analyses were repeated for the GD patients with leukopenia after glucocorticoid treatment (15 mg/day to 30 mg/day prednisone). RESULTS: The ELISA results showed that E-selectin levels were higher in GD patients with leukopenia than those with normal leukocytes (p < 0.05), but these levels decreased after glucocorticoid (prednisone) treatment (p < 0.05). No significant change in sICAM-1 levels was observed (p = 0.12). Correlation analysis showed that leukocyte count and E-selectin were negatively correlated (r = -0.778; p < 0.05). CONCLUSION: E-selectin may have an important function in GD with leukopenia, and glucocorticoids (prednisone) could decrease E-selectin level, which may be a new therapy target for GD with leukopenia.