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Ultra-high performance liquid chromatography-quadrupole-time of flight tandem mass spectrometry(UHPLC-Q-TOF-MS) was employed in this study to observe the effect of Huaihua Powder on the serum metabolites of mice with ulcerative colitis and reveal the mechanism of Huaihua Powder in the treatment of ulcerative colitis. The mouse model of ulcerative colitis was established by dextran sodium sulfate salt(DSS). The therapeutic effect of Huaihua Powder on ulcerative colitis was preliminarily evaluated based on the disease activity index(DAI), colon appearance, colon tissue morphology, and the content of inflammatory cytokines such as tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), and interleukin-1ß(IL-1ß). UHPLC-Q-TOF-MS was employed to profile the endogenous metabolites of serum samples in blank control group, model group, and low-, medium-, and high-dose Huaihua Powder groups. Multivariate analyses such as principal component analysis(PCA), partial least squares discriminant analysis(PLS-DA), and orthogonal partial least squares discriminant analysis(OPLS-DA) were performed for pattern recognition. Potential biomarkers were screened by Mass Profiler Professional(MPP) B.14.00 with the thresholds of fold change≥2 and P<0.05. The metabolic pathways were enriched by MetaboAnalyst 5.0. The results showed that Huaihua Powder significantly improved the general state and colon tissue morphology of mice with ulcerative colitis, reduced DAI, and lowered the levels of TNF-α, IL-6, and IL-1ß in serum. A total of 38 potential biomarkers were predicted to be related to the regulatory effect of Huaihua Powder, which were mainly involved in glycerophospholipid metabolism, glycine, serine, and threonine metabolism, mutual transformation of glucuronic acid, and glutathione metabolism. This study employed metabolomics to analyze the mechanism of Huaihua Powder in the treatment of ulcerative colitis, laying a foundation for the further research.
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Colitis Ulcerosa , Ratones , Animales , Colitis Ulcerosa/inducido químicamente , Colitis Ulcerosa/tratamiento farmacológico , Colitis Ulcerosa/metabolismo , Polvos , Factor de Necrosis Tumoral alfa/metabolismo , Interleucina-6/metabolismo , Metabolómica , Colon , Modelos Animales de Enfermedad , Biomarcadores , Sulfato de Dextran/metabolismo , Sulfato de Dextran/farmacología , Sulfato de Dextran/uso terapéuticoRESUMEN
Ovarian cancer is one of the most malignant gynecological tumors in the world, with a high fatality rate and resistance to chemotherapy. Much basic research has revealed that oxidative stress is involved in tumor occurrence and development processes and is associated with tumor progression as well as metastasis. In recent years, oxidative stress has been proven to have a close connection with malignant biological behavior in ovarian cancer, and this topic has garnered increasing attention. This article reviews the research aimed toward elucidating the relationship between oxidative stress and ovarian cancer.
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Resistencia a Antineoplásicos , Factor 2 Relacionado con NF-E2/metabolismo , Neoplasias Ováricas/tratamiento farmacológico , Neoplasias Ováricas/metabolismo , Elementos de Respuesta Antioxidante , Femenino , Humanos , Proteína 1 Asociada A ECH Tipo Kelch/metabolismo , Neoplasias Ováricas/genética , Neoplasias Ováricas/patología , Proteínas Proto-Oncogénicas p21(ras)/genética , Proteínas Proto-Oncogénicas p21(ras)/metabolismo , Receptor ErbB-2/genética , Receptor ErbB-2/metabolismo , Transducción de SeñalRESUMEN
Early cancer metastases often occur in cervical cancer (CC) patients, resulting in poor prognosis and poor therapeutic outcome after resection of primary cancer. Hence, there is a compelling requirement for elucidating the molecular mechanisms underlying the CC cell invasiveness. Recently, the role of microRNAs (miRNAs) and pituitary tumor-transforming gene 1 (Pttg1) in the carcinogenesis of CC has been reported. Nevertheless, the relationship between miRNAs and Pttg1 remains ill-defined. Here, we showed that the levels of miR-3666 were significantly decreased and the levels of zinc finger E-box binding homeobox 1 (ZEB1) and Pttg1 were significantly increased in the CC specimens from patients, compared to the paired non-tumor tissue. Moreover, the levels of miR-3666 and ZEB1 inversely correlated. Bioinformatics analyses showed that miR-3666 targeted the 3'-untranslated region (3'-UTR) of ZEB1 messenger RNA (mRNA) to inhibit its translation, which was confirmed by luciferase reporter assay. Moreover, Pttg1 overexpression inhibited miR-3666 and subsequently increased ZEB1 and cell invasion, while Pttg1 depletion increased miR-3666 and subsequently decreased ZEB1 and cell invasion. Together, our data suggest that Pttg1 may increase CC cell metastasis, possibly through miR-3666-regulated ZEB1 levels.
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OBJECTIVE: Macrophage apoptosis plays a key role in atherosclerotic plaque rupture. This study investigated the effects of recombinant human brain natriuretic peptide (BNP) on oxidised low-density lipoprotein (ox-LDL)-induced macrophage apoptosis and explored the underlying mechanism. METHODS: A model of ox-LDL-induced macrophage injury was established to evaluate the role of BNP. Flow cytometry was employed to detect apoptosis and changes in mitochondrial membrane potential (x0394;x03A8;m), and confocal microscopy was used to determine cellular reactive oxygen species (ROS) levels. Additionally, reverse transcription-polymerase chain reaction and colourimetry were used to detect the mRNA expression and activity, respectively, of superoxide dismutase (SOD) and malondialdehyde (MDA). RESULTS: Ox-LDL induced macrophage apoptosis in a concentration-dependent manner, and maximum apoptosis occurred at 100 µg/ml ox-LDL (45.62 ± 2.76 vs. 6.84 ± 1.94%; p < 0.05). Conversely, BNP suppressed macrophage apoptosis, with a maximal effect at 10-9 mol/l (18.56 ± 1.79%; p < 0.05). Compared with the control group, intracellular ROS levels increased, x0394;x03A8;m decreased, SOD mRNA expression and activity decreased and MDA mRNA expression and content increased in the 100-µg/ml ox-LDL group (527.30 ± 36.20 vs. 100.00 ± 0.00%, 3.01 ± 0.52 vs. 9.67 ± 0.51%, 0.53 ± 0.18 vs. 1.00 ± 0.00, 256.6 ± 8.20 vs. 355.8 ± 9.58 U/ml, 1.59 ± 0.23 vs. 1.00 ± 0.00 and 29.4 ± 1.68 vs. 5.94 ± 0.51 nmol/ml; p < 0.05); these effects were significantly counteracted by 10-9 mol/l BNP (237.30 ± 30.62%, 6.55 ± 1.57%, 0.90 ± 0.07, 310.4 ± 2.97 U/ml, 1.14 ± 0.10, 20.54 ± 1.55 nmol/ml; p < 0.05). CONCLUSION: BNP attenuates ox-LDL-induced macrophage apoptosis by suppressing oxidative stress and preventing x0394;x03A8;m loss.
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Apoptosis/efectos de los fármacos , Lipoproteínas LDL/farmacología , Macrófagos/patología , Péptido Natriurético Encefálico/farmacología , Estrés Oxidativo/efectos de los fármacos , Técnicas de Cultivo de Célula , Línea Celular , Supervivencia Celular/efectos de los fármacos , Humanos , Malondialdehído/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Proteínas Recombinantes/farmacología , Superóxido Dismutasa/metabolismoRESUMEN
Neuronal insulin signaling abnormalities have been associated with Alzheimer's disease (AD). However, the specificity of this association and its underlying mechanisms have been unclear. This study investigated the expression of abnormal serine phosphorylation of insulin receptor substrate 1 (IRS1) in 157 human brain autopsy cases that included AD, tauopathies, α-synucleinopathies, TDP-43 proteinopathies, and normal aging. IRS1-pS(616), IRS1-pS(312) and downstream target Akt-pS(473) measures were most elevated in AD but were also significantly increased in the tauopathies: Pick's disease, corticobasal degeneration and progressive supranuclear palsy. Double immunofluorescence labeling showed frequent co-expression of IRS1-pS(616) with pathologic tau in neurons and dystrophic neurites. To further investigate an association between tau and abnormal serine phosphorylation of IRS1, we examined the presence of abnormal IRS1-pS(616) expression in pathological tau-expressing transgenic mice and demonstrated that abnormal IRS1-pS(616) frequently co-localizes in tangle-bearing neurons. Conversely, we observed increased levels of hyperphosphorylated tau in the high-fat diet-fed mouse, a model of insulin resistance. These results provide confirmation and specificity that abnormal phosphorylation of IRS1 is a pathological feature of AD and other tauopathies, and provide support for an association between insulin resistance and abnormal tau as well as amyloid-ß.
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Enfermedad de Alzheimer/patología , Encéfalo/metabolismo , Proteínas Sustrato del Receptor de Insulina/metabolismo , Serina/metabolismo , Tauopatías/patología , Factores de Edad , Anciano , Anciano de 80 o más Años , Análisis de Varianza , Animales , Proteínas de Unión al ADN/metabolismo , Dieta Alta en Grasa/efectos adversos , Femenino , Humanos , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Persona de Mediana Edad , Fosforilación/genética , Proteinopatías TDP-43/patología , alfa-Sinucleína/metabolismo , Proteínas tau/genética , Proteínas tau/metabolismoRESUMEN
In the title salt, [Cd(C6H15NO3)2](C8H4O4), the Cd(2+) cation is coordinated by six O atoms and two N atoms from two tetra-dentate 2-[bis-(2-hy-droxy-eth-yl)amino]-ethanol ligands, displaying a distorted square-anti-prismatic coordination. The terephthalate dianion does not coordinate to the cation but is connected through Oâ¯H-O hydrogen bonds of medium strength to the complex cations, leading to a layered structure extending parallel to (100).
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Two 4,4'-[1,3-phenyl-enebis(-oxy)]dibenzoate anions bridge two 1,10-phenanthroline-chelated Zn(II) cations about a center of inversion to generate the dinuclear title compound, [Zn2(C20H12O6)2(C12H8N2)2]·2H2O. The geometry about the Zn(II) atom is a distorted octa-hedron. In the crystal, the mol-ecules are connected by classical O-Hâ¯O hydrogen bonds, weak C-Hâ¯O hydrogen bonds and C-Hâ¯π inter-actions, forming a three dimensional network. π-π stacking is also observed between aromatic rings of adjacent mol-ecules, centroid-centroid distances are 3.753â (2), 3.5429â (16) and 3.5695â (17)â Å.
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OBJECTIVE: To explore the clinical efficacy and safety of flumatinib mesylate produced in China in patients with newly diagnosed chronic myeloid leukemia in chronic phase (CML-CP). METHODS: 32 newly diagnosed CML-CP patients admitted to the Hematology Department of the Affiliated Hospital of Southwest Medical University from March 1, 2020 to March 31, 2022, who had never received any tyrosine kinase inhibitor (TKI) were included in the study. The patients were treated by flumatinib mesylate 600mg once daily. The hematologic, cytogenetic and molecular responses were assessed at 3-, 6- and 12-month, and adverse effects of the drug were evaluated. RESULTS: 31 patients were treated with flumatinib for≥3 months, of which 24 patients were treated for ≥6 months and 14 patients were treated for≥12 months. At 3rd month of treatment, 30 out of 31 patients achieved complete hematologic response (CHR); 24 patients underwent cytogenetic testing and 22 cases achieved major cytogenetic response(MCyR), of which 21 cases achieved complete cytogenetic response (CCyR); Among 25 patients who underwent molecular testing, 22 patients had BCR-ABLIS≤10%, including 10 patients with BCR-ABLIS≤0.1%, and 6 patients with BCR-ABLIS≤0.01%. At 6th month of treatment, 23 out of 24 patients achieved CHR; 17 patients underwent cytogenetic testing and all achieved CCyR; Among 23 patients who underwent molecular testing, 20 patients had BCR-ABLIS≤1%, including 16 patients with BCR-ABLIS≤0.1% and 12 patients with BCR-ABLIS≤0.01%. At 12nd month of treatment, all 14 patients achieved CHR and CCyR; Among them, 10 patients had BCR-ABLIS≤0.1%, including 9 patients with BCR-ABLIS≤0.01%. The grade â ¢/â £ leukopenia, thrombocytopenia and anemia rates in the patients were 13.3%, 20.0% and 3.3%, respectively. One patient stopped flumatinib therapy due to severe and persistent hematologic toxicity. The major non-hematologic adverse events were abnormal liver function (20%), diarrhea (10%), bone/joint pain (10%), muscle spasm (10%), rash (6.7%), acute kidney injury (6.7%) and nausea(3.3%), most of which were grade I-II. No patient experienced grade â £ non-hematologic adverse events. No drug toxicity-related death occurred. CONCLUSION: Flumatinib mesglate, as the first-line treatment for newly diagnosed CML-CP, can enable the patients to achieve early and deep molecular and cytogenetic responses, and shows good safety.
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Anemia , Antineoplásicos , Leucemia Mielógena Crónica BCR-ABL Positiva , Trombocitopenia , Humanos , Mesilato de Imatinib/uso terapéutico , Pirimidinas/farmacología , Proteínas de Fusión bcr-abl , Leucemia Mielógena Crónica BCR-ABL Positiva/tratamiento farmacológico , Benzamidas/uso terapéutico , Inhibidores de Proteínas Quinasas/uso terapéutico , Resultado del Tratamiento , Respuesta Patológica Completa , Mesilatos/uso terapéutico , Antineoplásicos/uso terapéuticoRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Colorectal cancer (CRC) belongs to the category of intestinal wind, anal ulcer, abdominal mass and other diseases in traditional Chinese medicine (TCM). Floris Sophorae Powder (F.S), is a classical prescription is recorded in Puji Benshi Fang for the treatment of intestinal carbuncle. It has been incorporated into the prescriptions for the treatment of intestinal diseases and achieved remarkable results in modern medicine. However, the mechanism of F.S in the treatment of colorectal cancer remains unclear and requires further study. AIM OF THE STUDY: To investigate F.S in treating CRC and clarify the underlying mechanism. MATERIALS AND METHODS: This study was based on Dextran Sulfate Sodium Salt (DSS) combined with Azoxymethane (AOM) induced CRC mouse model to clarify the pharmacological effects of F.S. The serum metabolomics was used to study the mechanism of action, and the chemical composition of F.S was found by UPLC-Q-TOF-MS. The rationality of serm metabolomics results was verified through the clinical target database of network pharmacology, and the upstream and downstream targets of related pathways were found. The mechanism pathway was verified by Western blot to clarify its mechanism of action. RESULTS: In vivo pharmacological experiments showed that F.S inhibited tumor growth and improved hematochezia. The vital signs of mice in the high-dose F.S group approached to those in the control group. A total of 43 differential metabolites were found to be significantly changed by serum metabolomics. F.S could modulate and recover most of the differential metabolites, which proved to be closely related to the KRAS/MEK-ERK signaling pathway. A total of 46 compounds in F.S were identified, and the rationality of serm metabolic pathway was verified by network pharmacology. Western blot results also verified that the expression of KRAS, E2F1, p-MEK and p-ERK were significantly decreased after F.S treatment. CONCLUSION: Classical prescription Floris Sophorae Powder treat colorectal cancer by regulating KRAS/MEK-ERK signaling pathway.
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Neoplasias Colorrectales , Medicamentos Herbarios Chinos , Animales , Ratones , Polvos/uso terapéutico , Proteínas Proto-Oncogénicas p21(ras)/metabolismo , Moduladores Selectivos de los Receptores de Estrógeno/uso terapéutico , Transducción de Señal , Quinasas de Proteína Quinasa Activadas por Mitógenos/metabolismo , Neoplasias Colorrectales/metabolismo , Medicamentos Herbarios Chinos/farmacología , Medicamentos Herbarios Chinos/uso terapéuticoRESUMEN
OBJECTIVE: To explore the alteration of collagen ultrastructure and content in uterine ligaments and paraurethral tissue and explore whether the alteration may contribute to stress urinary incontinence (SUI) and pelvic organ prolapse (POP). METHODS: The cardinal ligament, uterosacral ligament and paraurethral tissue samples were obtained from 90 subjects undergoing hysterectomy. Collagen ultrastructure was examined with transmission electron microscopy. And collagen content and expression of vasoactive intestinal peptide (VIP) were examined with immunohistochemistry. RESULTS: The smooth muscle fascicles were thinner in the patients of SUI and POP. Arrangement of smooth muscle fascicles was disorderly. Fibroblast was metabolically active. The mean collagen fibril diameters in the SUI and POP groups were larger than that in the control group (P < 0.01). The mean contents of collagen I and III in the SUI and POP groups were lower than that in the control group (P < 0.01). The expression of VIP was lower (P < 0.05). CONCLUSION: Predominance of collagen degradation during tissue repair may contribute to and promote POP and SUI. The decrease of VIP might be related with nerve damage or degeneration to cause or accelerate the progress of pelvic organ prolapse.
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Colágeno/ultraestructura , Prolapso de Órgano Pélvico/patología , Incontinencia Urinaria de Esfuerzo/patología , Péptido Intestinal Vasoactivo/metabolismo , Colágeno/metabolismo , Femenino , Humanos , Persona de Mediana Edad , Diafragma Pélvico/patología , Prolapso de Órgano Pélvico/metabolismo , Incontinencia Urinaria de Esfuerzo/metabolismoRESUMEN
OBJECTIVE: To explore the effect of recombinant human thrombopoietin (rhTPO) on hematopoietic reconstruction in allogeneic hematopoietic stem cell transplantation (allo-HSCT) model. METHODS: The C57BL/6 mice were employed as the donors, and BALB/c mice as recipients. The bone marrow mononuclear cells of the donor mice were extracted and pretreated, which then were injected with 5×106 per mouse through the tail vein of the recipient to establish an allo-HSCT model. The implantation of hematopoietic stem cells in the recipient mice was detected by flow cytometry on the 28th day after transplantation. Next, the successfully modeled recipient mice were randomly divided into experimental group and control group. The rhTPO was injected into mice in the experimental group on the first day after transplantation, while the saline was injected into mice in the control group. Both groups were injected for 14 consecutive days. The peripheral blood and bone marrow hematopoiesis of the two groups were observed on day 1, 3, 7, 14, and 21 after transplantation. RESULTS: The expression rate of H-2Kb in the bone marrow of recipient mice was 43.85% (>20%) on the 28th day after transplantation, which indicated that the recipient mice were successfully chimerized. Meanwhile, counts of PLTs on the day 3, 7, 14, and 21 after transplantation in the experimental group were higher than those in the control group with statistical significances (P<0.05). In addition, hematopoietic function of bone marrow was suppressed in both groups on day 1, 3 and 7 after transplantation, but hematopoietic bone marrow hyperplasia was better in the experimental group than in the control group. On day 14 and 21 after transplantation, the hematopoietic function of bone marrow in the two groups was recovered, and the experimental group showed more obvious than the control group. CONCLUSION: rhTPO can effectively stimulate the production of PLTs and facilitate the recovery of white blood cells and hemoglobin after allo-HSCT, and promote hematopoietic recovery and reconstitution of bone marrow.
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Trasplante de Células Madre Hematopoyéticas , Trombopoyetina , Humanos , Animales , Ratones , Ratones Endogámicos C57BL , Células Madre Hematopoyéticas , Médula Ósea , Proteínas Recombinantes , Ratones Endogámicos BALB CRESUMEN
OBJECTIVE: To explore the feasibility and safety of mdr1 gene transferred into placenta derived mesenchymal stem cells (P-MSCs) by reconstructed retroviral vector. METHODS: Human P-MSCs were isolated and expanded by Percoll density gradient and then transduced repeatedly by reconstructed retroviral vector containing mdr1 gene. The transfection and expression of mdr1 gene was detected by reverse transcription-polymerase chain reaction (RT-PCR) and fluorescence-activated cell sorting (FACS). Meanwhile, the biological features of mdr1-MSCs were identified and analyzed. RESULTS: The expression of mdr1mRNA was found in transfected cells. The expression of P-glycoprotein (P-gp) encoded by mdr1 gene was (27.6 ± 5.1)% in the transfected P-MSCs cells versus (0.4 ± 0.1)% in the non-transfected P-MSCs cells (t = 14.291, P < 0.01). The percent of P-MSCs at quiescent phase (G0/G1 phase) was around 95.40% and it was in accord with the characterization of stem cells. The mdr1-MSCs exhibited typical ultrastructures of low-differentiated stem cells. Moreover, they still retained the potency of adipogenic and osteogenic differentiation in the presence of appropriate conditioned media. CONCLUSION: A stable expression of P-gp may be obtained by reconstructed retroviral-mediated transfection in vitro. And transfected MSCs retain the characteristics of stem cells.
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Resistencia a Múltiples Medicamentos/genética , Genes MDR , Células Madre Mesenquimatosas/citología , Placenta/citología , Transfección , Miembro 1 de la Subfamilia B de Casetes de Unión a ATP/genética , Células Cultivadas , Femenino , Vectores Genéticos , Humanos , EmbarazoRESUMEN
RATIONALE AND OBJECTIVES: The purpose of this study was to explore conventional MRI features that can accurately differentiate central nervous system embryonal tumor, not otherwise specified (CNS ETNOS) from glioblastoma (GBM) in adults. MATERIALS AND METHODS: Preoperative conventional MRI images of 30 CNS ETNOS and 98 GBMs were analyzed by neuroradiologists retrospectively to identify valuable MRI features. Five blinded neuroradiologists independently reviewed all these MRI images, and scored MRI features on a five-point scale. Kendall's coefficient of concordance was used to measure inter-rater agreement. Diagnostic value was assessed by the area under the curve (AUC) of receiver operating curve, and sensitivity and specificity were also calculated. RESULTS: Seven MRI features, including isointensity on T1WI, T2WI, and FLAIR, ill-defined margin, severe peritumoral edema, ring enhancement, and broad-based attachment sign, were helpful for the differential diagnosis of these two entities. Among these features, ring enhancement showed the highest inter-rater concordance (0.80). Ring enhancement showed the highest AUC value (0.79), followed by severe peritumoral edema (0.67). The combination of seven features showed the highest AUC value (0.86), followed by that of three features (ill-defined margin, severe peritumoral edema, and ring enhancement) (0.83). CONCLUSION: Enhancement pattern, peritumoral edema, and margin are valuable for the discrimination between CNS ETNOS and GBM in adults.
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Neoplasias Encefálicas , Glioblastoma , Adulto , Neoplasias Encefálicas/diagnóstico por imagen , Neoplasias Encefálicas/patología , Sistema Nervioso Central/patología , Glioblastoma/diagnóstico por imagen , Glioblastoma/patología , Humanos , Imagen por Resonancia Magnética/métodos , Márgenes de Escisión , Estudios RetrospectivosRESUMEN
OBJECTIVE: Olfactory dysfunction is common in Alzheimer disease (AD) and other neurodegenerative diseases. Paired helical filament (PHF)-tau, alpha-synuclein, and amyloid-beta lesions occur early and severely in cerebral regions of the olfactory system, and they have also been observed in olfactory epithelium (OE). However, their frequency, abundance, and disease specificity, and the relationships of OE pathology to brain pathology have not been established. METHODS: We investigated the pathological expression of amyloid-beta, PHFtau, alpha-synuclein, and TDP-43 in postmortem OE of 79 cases with AD, 63 cases with various other neurodegenerative diseases, and 45 neuropathologically normal cases. RESULTS: Amyloid-beta was present as punctate and small patchy aggregates in 71% of AD cases, compared with 22% of normal cases and 14% of cases with other diseases, and in greater amounts in AD than in either of the other 2 diagnostic categories. PHFtau was evident in dystrophic neurites in 55% of cases with AD, 34% with normal brains, and 39% with other neurodegenerative diseases, also at higher densities in AD. alpha-Synuclein was present in dystrophic neurites in 7 cases, 6 of which also had cerebral Lewy bodies. Pathological TDP-43 inclusions were not observed in the OE in any cases. Amyloid-beta and to a lesser degree, PHFtau ratings in OE significantly correlated with cortical Abeta and PHFtau lesion ratings in the brain. INTERPRETATION: These data demonstrate that AD pathology in the OE is present in the majority of cases with pathologically verified AD and correlates with brain pathology. Future work may assess the utility of amyloid-beta and PHFtau measurement in OE as a biomarker for AD.
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Enfermedad de Alzheimer/patología , Péptidos beta-Amiloides/metabolismo , Ovillos Neurofibrilares/metabolismo , Mucosa Olfatoria/metabolismo , Mucosa Olfatoria/patología , Proteínas tau/metabolismo , Anciano , Anciano de 80 o más Años , Análisis de Varianza , Proteínas de Unión al ADN/metabolismo , Femenino , Humanos , Cuerpos de Lewy/metabolismo , Cuerpos de Lewy/patología , Masculino , Ovillos Neurofibrilares/patología , Estadística como Asunto , alfa-Sinucleína/metabolismoRESUMEN
Animal models provide compelling evidence that chronic stress is associated with biochemical and morphological changes in the brain, many of which are mediated by corticosterone, a principal glucocorticoid synthesized in the rodent adrenal cortex and secreted in response to stress. To better characterize the effects of chronic corticosterone at the synaptic and subsynaptic level, we implanted three-month-old male C57B/6 mice with 2 × 5 mg corticosterone pellets (CORT group, n = 14), 21 day release formulation (20 mg/kg/day dose) or placebo pellets (Placebo group, n = 14), 21-day release formulation. After 20 days, brains were removed. One hemisphere was frozen for biochemical analysis by synaptosomal fractionation with Western blotting, and the other hemisphere was fixed for immunohistochemistry. Localization and expression levels for PSD-95, NR1, and synaptopodin proteins were assessed. Biochemical analysis revealed lower protein levels of PSD-95 (32% decrease, P < 0.001), NR1 (47%, P = 0.01), and synaptopodin (65%, P < 0.001) in the postsynaptic density subsynaptic fraction of the CORT group. Optical densitometry in immunohistochemically labeled sections also found lower levels of PSD-95 in synaptic fields of the dentate gyrus (PSD-95, 33% decrease, P < 0.001; NR1, 31%, P < 0.001; synaptopodin, 40%, P < 0.001) and the CA3 stratum lucidum (36%, P < 0.001, 40%, P < 0.001, and 35%, P < 0.001) of the CORT group. While mechanistic relationships for these changes are not yet known, we speculate that synaptopodin, which is involved in regulation of spine calcium kinetics and posttranslational modification and transport of locally synthesized proteins, may play an important role in the changes of PSD-95 and NR1 protein levels and other synaptic alterations.
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Encéfalo/metabolismo , Corticosterona/metabolismo , Guanilato-Quinasas/metabolismo , Proteínas de la Membrana/metabolismo , Proteínas de Microfilamentos/metabolismo , Densidad Postsináptica/metabolismo , Receptores de N-Metil-D-Aspartato/metabolismo , Animales , Western Blotting , Homólogo 4 de la Proteína Discs Large , Inmunohistoquímica , Masculino , Ratones , Ratones Endogámicos C57BL , Densidad Postsináptica/químicaRESUMEN
In the title compound, {[Zn(C(8)H(5)N(2)O(2)S)(2)]·3H(2)O}(n), the Zn(II) atom, lying on a twofold rotation axis, is four-coordinated by two S atoms and two O atoms from four 2-sulfido-1H-benzimidazol-3-ium-5-carboxyl-ate (H(2)mbidc) ligands in a distorted tetra-hedral geometry. Two H(2)mbidc ligands bridge two Zn(II) atoms, generating a double-chain along [[Formula: see text]01]. Adjacent chains are linked by N-Hâ¯O and O-Hâ¯O hydrogen bonds, forming a three-dimensional supra-molecular network. One of the two water molecules also lies on a twofold rotation axis.
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Progressive synaptic degeneration and neuron loss are major structural correlates of cognitive impairment in Alzheimer's disease (AD). The mechanisms by which synaptic degeneration in AD occurs have not been established. The activation of proteins within the caspase family has been implicated in AD-associated neurodegeneration, and synaptically localized caspase activity could play a role in the synaptic degeneration and loss found in AD. We used synaptosomal fractionation with Western blotting and immunohistochemistry to examine the anatomical, subcellular, and subsynaptic expression patterns of caspase 3 in both the anterior cingulate cortex and hippocampus of control and AD patients. In both control and AD cases, there was a selective enrichment of caspase- 3 at synapses, particularly in the postsynaptic density (PSD) fractions. Compared with controls, AD patients exhibited significant increases in synaptic procaspase- 3 and active caspase-3 expression levels that were most evident in the PSD fractions. These data demonstrate for the first time the preferential localization and increase of caspase-3 in the PSD fractions in AD and suggest an important role for caspase 3 in synapse degeneration during disease progression.
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Enfermedad de Alzheimer/enzimología , Caspasa 3/metabolismo , Sinapsis/enzimología , Anciano , Enfermedad de Alzheimer/patología , Western Blotting , Núcleo Celular/enzimología , Núcleo Celular/patología , Citosol/enzimología , Citosol/patología , Activación Enzimática , Femenino , Hipocampo/enzimología , Hipocampo/patología , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana Edad , Transporte de Proteínas , Sinaptosomas/enzimología , Sinaptosomas/patología , Extractos de TejidosRESUMEN
OBJECTIVE: To transfer multidrug resistance gene (mdr1) into human placental mesenchymal stem cells (P-MSCs) by retroviral vector and assess the effects of mdr1 gene transduction upon biological features of P-MSCs. METHODS: Human P-MSCs were isolated from trypsin-digested term placentas and then transduced by reconstructed retroviral vector containing mdr1 gene and green fluorescent protein (GFP) reporter gene. Flow cytometric analysis was employed to determine the immunophenotypes of transfected P-MSCs. And the proliferation and cell cycle were detected by methyl thiazolyl tetrazolium and propidium iodide staining. Ultrastructures of transfected P-MSCs were observed and different induction conditions used to direct the cells to differentiate into adipocytes and osteoblasts. RESULTS: The transfected P-MSCs still expressed stem cell markers such as CD29, CD44 and CD73. The mean cumulative time of population doubling was 23.9 hours. The cellular cycle retained the proliferative characterization of stem cells. Ultrastructural features of transfected P-MSCs included increased surface microvilli, abundant mitochondria and slightly swollen rough endoplasmic reticulum. Furthermore these transfected cells demonstrated osteogenic and adipogenic differentiation potentials under appropriate conditions. CONCLUSION: The mdr1 gene transduction by retroviral vector in vitro has no significant effect upon biological characteristics of P-MSCs. It might provide experimental references for the application of P-MSCs in high-dose tumor chemotherapy.
Asunto(s)
Miembro 1 de la Subfamilia B de Casetes de Unión a ATP/genética , Resistencia a Múltiples Medicamentos/genética , Células Madre Mesenquimatosas/citología , Placenta/citología , Transfección , Diferenciación Celular , Resistencia a Antineoplásicos/genética , Femenino , Técnicas de Transferencia de Gen , Vectores Genéticos , Humanos , EmbarazoRESUMEN
OBJECTIVE: To investigate the effects of ovarian carcinoma cells on the differentiation, maturation, and function of the dendritic cells (DC). METHODS: Human epithelial ovarian carcinoma cells were isolated from specimens of ovarian carcinoma from 12 patients obtained during operation, and cultured. Peripheral blood samples were collected from these patients. DC were isolated and co-cultured with recombinant human tumor necrosis factor (TNF)-alpha to promote their maturation. Immunostaining and flow cytometry (FC) were used to detect the DC specific marker CD1a, maturation marker CD83, co-stimulating factors CD86 and human leucocyte antigen (HLA)-DR. FITC-labeled glucan was added and FC was used to detect the phagocytic function. T cells were co-cultured with immature and mature DC of different concentrations respectively for 96 h, 18 h before the end of culture 3H-TdR was added, liquid scintillation counter was used to measure the level of count per minute (cpm) Ovarian carcinoma cells and peripheral mononuclear cells were put into Transwell to contact each other directly or indirectly, TNF-alpha was add to promote the maturation, and then FC was used to detect the CD1a, CD83, CD86, and HLA-DR levels, and the glucan uptake level. ELISA was used to detect the levels of IL-12, IL-10, and interferon (IFN)-gamma, and Western blotting was used to detect the p-38 protein and phosphorylated p-38 protein in the supernatant. RESULTS: After stimulation of TNF-alpha the expression; levels of CD1a, CD86, CD83, and HLA-DR of the DCs were remarkably increased. Immature DCs showed a strong ability at phagocytosis of glucan, and this ability was decreased by the stimulation of TNF-alpha. The proliferative function of mature DCs on allogenic T lymphocytes was significantly increased dose-dependently (P<0.05). Co-culture with ovarian carcinoma cells decreased the expression levels of CD1a, CD86, and HLA-DR, increased the expression level of CD83, and decreased the ability in up taking glucan by 43.05%. The proliferative function of the DC cultured in direct contact with ovarian carcinoma cells was decreased by 56.35%, and such inhibitory function of the DCs cultured not in direct contact with ovarian carcinoma cells was weaker, and the levels of CD1a, CD86, CD83, and HLA-DR expressed by the DC cultured not in direct contact with ovarian carcinoma cells was between those of the DC cultured in direct contact with ovarian carcinoma cells and those of the DC undergoing routine culture. The levels of IL-12 and phosphorylated p-38 protein were decreased in the supernatant of the DC co-cultured with ovarian carcinoma cells. CONCLUSION: Ovarian carcinoma cells suppress the differentiation of monocyte into DC, negatively regulates the phagocytotic and antaean managing functions of DC, thus the DC can not effectively stimulate the proliferation of T lymphocytes and inhibit the activity of cytotoxic T lymphocytes. That may be one of the underlying mechanisms of immune escape of tumor and immune tolerance of T lymphocytes.
Asunto(s)
Diferenciación Celular , Células Dendríticas/inmunología , Tolerancia Inmunológica , Línea Celular Tumoral , Técnicas de Cocultivo , Células Dendríticas/citología , Femenino , Humanos , Escape del TumorRESUMEN
In the title compound, [Cd(C(16)H(10)O(4))(C(14)H(8)N(4))(H(2)O)]·0.5C(3)H(7)NO, the Cd(II) atom is six-coordinated by two N atoms from one pyrazino[2,3-f][1,10]phenanthroline ligand, three carboxyl-ate O atoms from two different 4,4'-ethyl-enedibenzoate ligands, and one water mol-ecule in a distorted octa-hedral environment. The two 4,4'-ethyl-enedibenzoate dianions are located on inversion centres bridging two neighboring Cd(II) centres. O-Hâ¯O hydrogen-bonding inter-actions further stabilize the crystal structure. The DMF molecule is equally disordered about a center of inversion.