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1.
Neuropsychiatr Dis Treat ; 18: 1917-1925, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36065387

RESUMO

Purpose: Post-stroke depression (PSD) is the most common psychiatric sequelae of stroke. Numerous studies revealed that event-related potentials (ERP) can reflect depression severity to a certain extent, while there is almost no research on depression after hemorrhagic stroke. Therefore, we employed a prospective cross-sectional study to explore the relationship between ERP and depression after hemorrhagic stroke. Methods: A total of 74 patients with intracranial hemorrhage were included in this study. Neurological deficits were evaluated using the National Institutes of Health Stroke Scale (NIHSS) on admission. Depression severity and cognitive impairment were measured using the 17-item Hamilton Depression Scale (HAMD-17) and the Chinese version of the Montreal Cognitive Assessment (MoCA) after two weeks of treatment. All patients were conducted auditory Oddball paradigm for event-related potential mismatch negativity (MMN) and P300. Results: In total, 36 patients were diagnosed with PSD at the two weeks of treatment, for a percentage of 48.6%. Depression severity of ICH patients correlated positively with both the latency of MMN (r = 0.376, P = 0.001) and P300 (r = 0.325, P = 0.005), and correlated negatively with both the amplitude of MMN (r=-0.385, P = 0.001) and P300 (r=-0.311, P = 0.007). Depression severity was negatively correlated with cognitive function after hemorrhagic stroke (r=-0.347, P = 0.002). Conclusion: The latency and amplitude of MMN and P300 can well reflect the degree of depression after hemorrhagic stroke, which may help in the early diagnosis and effective treatment of PSD.

2.
Psychol Med ; 52(13): 2651-2660, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-33298220

RESUMO

BACKGROUND: This study aimed to explore the impacts of COVID-19 outbreak on mental health status in general population in different affected areas in China. METHODS: This was a comparative study including two groups of participants: (1) general population in an online survey in Ya'an and Jingzhou cities during the COVID-19 outbreak from 10-20 February 2020; and (2) matching general population selected from the mental health survey in Ya'an in 2019 (from January to May 2019). General Health Questionnaire (GHQ-12), Self-rating Anxiety Scale (SAS), and Self-rating Depression Scale (SDS) were used. RESULTS: There were 1775 participants (Ya'an in 2019 and 2020: 537 respectively; Jingzhou in 2020: 701). Participants in Ya'an had a significantly higher rate of general health problems (GHQ scores ⩾3) in 2020 (14.7%) than in 2019 (5.2%) (p < 0.001). Compared with Ya'an (8.0%), participants in Jingzhou in 2020 had a significantly higher rate of anxiety (SAS scores ⩾50, 24.1%) (p < 0.001). Participants in Ya'an in 2020 had a significantly higher rate of depression (SDS scores ⩾53, 55.3%) than in Jingzhou (16.3%) (p < 0.001). The risk factors of anxiety symptoms included female, number of family members (⩾6 persons), and frequent outdoor activities. The risk factors of depression symptoms included participants in Ya'an and uptake self-protective measures. CONCLUSIONS: The prevalence of psychological symptoms has increased sharply in general population during the COVID-19 outbreak. People in COVID-19 severely affected areas may have higher scores of GHQ and anxiety symptoms. Culture-specific and individual-based psychosocial interventions should be developed for those in need during the COVID-19 outbreak.


Assuntos
COVID-19 , Humanos , Feminino , Saúde Mental , SARS-CoV-2 , Depressão/epidemiologia , Ansiedade/psicologia , Surtos de Doenças , Inquéritos e Questionários , China/epidemiologia
3.
Medicine (Baltimore) ; 100(21): e26050, 2021 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-34032732

RESUMO

ABSTRACT: To investigate the clinicopathological features and outcomes of primary IgA nephropathy with nephrotic-range proteinuria in Chinese children. Patients with biopsy-proven IgA nephropathy and nephrotic-range proteinuria between January 2011 and December 2017 were included, and their proteinuria and renal function were followed up. A total of 90 patients were enrolled, and 21.1% (19/90) of them had decreased renal function at diagnosis. Complete remission, partial remission, and no response of proteinuria occurred in 88.6% (70/79), 10.1% (8/79), and 1.3% (1/79), respectively, of the 79 patients who were followed up for 6 to 104 months. 73.7% (14/19) of the patients with decreased renal function at diagnosis recovered to normal level while 26.3% (5/19) of them did not recover or progressed to end-stage renal disease. Two patients with normal renal function at diagnosis progressed to renal insufficiency during follow-up period. By multivariate analysis, the risk for renal function deterioration was significantly higher in the partial remission and no response groups than in the complete remission group. Remission of proteinuria was important for improving renal prognosis in children with IgA nephropathy and nephrotic-range proteinuria. The outcomes for pediatric patients appeared to be better than that reported in adults.


Assuntos
Glomerulonefrite por IGA/tratamento farmacológico , Glucocorticoides/uso terapêutico , Imunossupressores/uso terapêutico , Falência Renal Crônica/epidemiologia , Proteinúria/epidemiologia , Adolescente , Biópsia , Criança , Pré-Escolar , China/epidemiologia , Quimioterapia Combinada/métodos , Feminino , Seguimentos , Mesângio Glomerular/patologia , Glomerulonefrite por IGA/complicações , Glomerulonefrite por IGA/imunologia , Glomerulonefrite por IGA/patologia , Humanos , Falência Renal Crônica/diagnóstico , Falência Renal Crônica/imunologia , Falência Renal Crônica/urina , Masculino , Proteinúria/diagnóstico , Proteinúria/imunologia , Proteinúria/urina , Indução de Remissão/métodos , Estudos Retrospectivos , Resultado do Tratamento
4.
Ann Transl Med ; 8(18): 1158, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-33241007

RESUMO

BACKGROUND: To evaluate the role of high-resolution computed tomography (HRCT) in the diagnosis of 2019 novel coronavirus (2019-nCoV) pneumonia and to provide experience in the early detection and diagnosis of 2019-nCoV pneumonia. METHODS: Seventy-two patients confirmed to be infected with 2019-nCoV from multiple medical centers in western China were retrospectively analyzed, including epidemiologic characteristics, clinical manifestations, laboratory findings and HRCT chest features. RESULTS: All patients had lung parenchymal abnormalities on HRCT scans, which were mostly multifocal in both lungs and asymmetric in all patients, and were mostly in the peripheral or subpleural lung regions in 52 patients (72.22%), in the central lung regions in 16 patients (22.22%), and in both lungs with "white lung" manifestations in 4 patients (5.56%). Subpleural multifocal consolidation was a predominant abnormality in 38 patients (52.78%). Ground-glass opacity was seen in 34 patients (47.22%). Interlobular septal thickening was found in 18 patients, 8 of whom had only generally mild thickening with no zonal predominance. Reticulation was seen in 8 patients (11.11%), and was mild and randomly distributed. In addition, both lungs of 28 patients had 2 or 3 CT imaging features. Out of these 72 patients, 36 were diagnosed as early stage, 32 patients as progressive stage, and 4 patient as severe stage pneumonia. Moreover, the diagnostic accuracy of HRCT features combined with epidemiological history was not significantly different from the detection of viral nucleic acid (all P >0.05). CONCLUSIONS: The HRCT features of 2019-nCoV pneumonia are characteristic to a certain degree, which when combined with epidemiological history yield high clinical value in the early detection and diagnosis of 2019-nCoV pneumonia.

5.
Aging (Albany NY) ; 12(18): 17902-17920, 2020 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-32976115

RESUMO

Non-small cell lung cancer (NSCLC) is a type of refractory malignant lung cancer with a high rate of metastasis and mortality. Currently, long non-coding RNA (lncRNA) SBF2 Antisense RNA 1 (SBF2-AS1) is considered as a biomarker for a variety of tumors. However, the function of SBF2-AS1 in the growth and metastasis of NSCLC needs to be further studied. In this study, we revealed that SBF2-AS1 was overexpressed in NSCLC tissues compared with that in normal tissues. SBF2-AS1 silencing restrained the growth and aggressive phenotypes of NSCLC cell in vitro. Consistently, SBF2-AS1 knockdown hindered the growth of NSCLC cell in nude mice. The following luciferase reporter gene assay and RNA immunoprecipitation (RIP) assay suggested the relationship between miR-338-3p and SBF2-AS1. The rescue experiments showed that miR-338-3p inhibitor abolished SBF2-AS1 silencing caused inhibition on the growth, migration and invasiveness of NSCLC cell. The luciferase reporter assay and immunoblotting assay validated that A Disintegrin and Metalloprotease 17 (ADAM17) was a target of miR-338-3p. In addition, SBF2-AS1 positively regulated the level of ADAM17 through sponging for miR-338-3p. Finally, we revealed that SBF2-AS1 contributed to the proliferation and metastatic phenotypes of NSCLC cell via regulating miR-338-3p/ADAM17 axis.

6.
J Nanosci Nanotechnol ; 10(7): 4667-73, 2010 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-21128476

RESUMO

This paper presents an adaptive neuro-fuzzy approach based on first order function of fuzzy model for establishing the relationship between control factors and thin films properties of TiN/ZrN coatings on Si(100) wafer substrates. A statistical model was designed to explore the space of the processes by an orthogonal array scheme. Eight control factors of closed unbalance magnetron sputtering system were selected for modeling the process, such as interlayer material, argon and nitrogen flow rate, titanium and zirconium target current, rotation speed, work distance, and bias voltage. Analysis of variance (ANOVA) was carried out for determining the influence of control factors. In this study, with the application of ANOVA, the smallest effect of control factors was eliminated. The adaptive neuro-fuzzy inference system (ANFIS) was applied as a tool to model the deposited process with five significant control factors. The experimental results show that ANFIS demonstrates better accuracy than additive model for the film hardness. The root mean square error between prediction values and experimental values were archived to 0.04.

7.
J Zhejiang Univ Sci B ; 10(7): 512-21, 2009 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-19585669

RESUMO

The dense granule protein 4 (GRA4) is a granular protein from Toxoplasma gondii, and is a candidate for vaccination against this parasite. In this study, the plasmid pcDNA3.1-GRA4 (pGRA4), encoding for the GRA4 antigen, was incorporated by the dehydration-rehydration method into liposomes composed of 16 mmol/L egg phosphatidylcholine (PC), 8 mmol/L dioleoyl phosphatidylethanolamine (DOPE), and 4 mmol/L 1,2-diodeoyl-3-(trimethylammonium) propane (DOTAP). C57BL/6 mice and BALB/c mice were immunized intramuscularly three times with liposome-encapsulated pGRA4 to determine whether DNA immunization could elicit a protective immune response to T. gondii. Enzyme-linked immunosorbent assay (ELISA) of sera from immunized mice showed that liposome-encapsulated pGRA4 generated high levels of IgG antibodies to GRA4. Production of primary interferon (IFN)-gamma and interleukin (IL)-2 in GRA4-stimulated splenocytes from vaccinated mice suggested a modulated Th1-type response. 72.7% of C57BL/6 mice immunized with liposome-encapsulated pGRA4 survived the challenge with 80 tissue cysts of ME49 strain, whereas C57BL/6 mice immunized with pGRA4 had only a survival rate of 54.5%. When immunized BALB/c mice were intraperitoneally challenged with 10(3) tachyzoites of the highly virulent RH strain, the survival time of mice immunized with liposome-encapsulated pGRA4 was markedly longer than that of other groups. Our observations show that liposome-encapsulated pGRA4 enhanced the protective effect against infection of T. gondii.


Assuntos
Lipossomos/química , Proteínas de Protozoários/genética , Proteínas de Protozoários/imunologia , Toxoplasmose/imunologia , Toxoplasmose/prevenção & controle , Vacinas de DNA/administração & dosagem , Vacinas de DNA/química , Animais , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Resultado do Tratamento
8.
Cell Transplant ; 16(6): 579-85, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-17912949

RESUMO

Human mesenchymal stem cells (MSCs) are multipotential and are detected in bone marrow (BM), adipose tissue, placenta, and umbilical cord blood (UCB). In this study, we examined the ability of UCB-derived MSCs (UCB-MSCs) to support ex vivo expansion of hematopoietic stem/progenitor cells (HSPCs) from UCB and the engraftment of expanded HSPCs in NOD/SCID mice. The result showed that UCB-MSCs supported the proliferation and differentiation of CD34+ cells in vitro. The number of expanded total nucleated cells (TNCs) in MSC-based culture was twofold higher than cultures without MSC (control cultures). UCB-MSCs increased the expansion capabilities of CD34+ cells, long-term culture-initiating cells (LTC-ICs), granulocyte-macrophage colony-forming cells (GM-CFCs), and high proliferative potential colony-forming cells (HPP-CFCs) compared to control cultures. The expanded HSPCs were transplanted into lethally irradiated NOD/SCID mice to assess the effects of expanded cells on hematopoietic recovery. The number of white blood cells (WBCs) in the peripheral blood of mice transplanted with expanded cells from both the MSC-based and control cultures returned to pretreatment levels at day 25 posttransplant and then decreased. The WBC levels returned to pretreatment levels again at days 45-55 posttransplant. The level of human CD45+ cell engraftment in primary recipients transplanted with expanded cells from the MSC-based cultures was significantly higher than recipients transplanted with cells from the control cultures. Serial transplantation demonstrated that the expanded cells could establish long-term engraftment of hematopoietic cells. UCB-MSCs similar to those derived from adult bone marrow may provide novel targets for cellular and gene therapy.


Assuntos
Antígenos CD34/análise , Sangue Fetal/citologia , Sobrevivência de Enxerto , Transplante de Células-Tronco Hematopoéticas , Células-Tronco Hematopoéticas/citologia , Células-Tronco Mesenquimais/citologia , Adulto , Animais , Proliferação de Células , Técnicas de Cocultura , Transplante de Células-Tronco de Sangue do Cordão Umbilical , Feminino , Fatores de Crescimento de Células Hematopoéticas/farmacologia , Células-Tronco Hematopoéticas/fisiologia , Humanos , Separação Imunomagnética/classificação , Imunofenotipagem , Contagem de Leucócitos , Masculino , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/fisiologia , Camundongos , Camundongos Endogâmicos NOD , Camundongos SCID , Gravidez , Quimera por Radiação , Transplante Heterólogo
9.
Cell Physiol Biochem ; 20(1-4): 205-12, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-17595529

RESUMO

UNLABELLED: This study was supported by grants of New Ideas Capability for Backbone Teachers in Universities of Heilongjiang and of Scientific Research foundation in Qiqihar Medical College. BACKGROUND/AIMS: Ulcer recurrence and poor healing may be critically important to the development of serious gastrointestinal complications in patients with long-term non-steroid anti-inflammatory drugs (NSAIDs). The present study is to investigate the effects of aspirin on ulcer recurrence and healing quality and to explore the mechanism. METHODS: Gastric ulcers were induced with acetic acid in rats; aspirin was administrated by gavage from day 25 to day 54 after ulcer induction. The gastric juice volume, pH, gastric mucus, gastric mucosal blood flow (GMBF) and prostaglandin E(2) (PGE(2)) were measured. The mRNA transcription of cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) were analyzed with RT-PCR and protein expression with Western blot. RESULTS: The gastric juice volume was significantly increased in aspirin group compared with those of fasting or saline control groups (P<0.01); while the pH, mucus, GMBF and PGE(2) were significantly decreased in aspirin treated rats compared with those of other two groups (P<0.01). COX-2, evaluated with mRNA and protein expression, was significantly augmented in aspirin group compared with others. The quality of ulcer healing (QOUH) in Aspirin group was poorer than that of fasting or saline control groups. CONCLUSIONS: Aspirin enhance the recurrence of gastric ulcer. The inhibition of cycloxygenase, mucus secretion and mucosal blood flow may be involved. Aspirin also impair the quality of ulcer healing.


Assuntos
Ácido Acético/toxicidade , Anti-Inflamatórios não Esteroides/toxicidade , Aspirina/toxicidade , Úlcera Gástrica/induzido quimicamente , Animais , Sequência de Bases , Ciclo-Oxigenase 1/genética , Ciclo-Oxigenase 2/genética , Primers do DNA/genética , Dinoprostona/metabolismo , Suco Gástrico/metabolismo , Mucosa Gástrica/irrigação sanguínea , Mucosa Gástrica/efeitos dos fármacos , Mucosa Gástrica/patologia , Concentração de Íons de Hidrogênio , Masculino , Proteínas de Membrana/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Recidiva , Fluxo Sanguíneo Regional/efeitos dos fármacos , Úlcera Gástrica/genética , Úlcera Gástrica/patologia , Úlcera Gástrica/fisiopatologia
10.
J Zhejiang Univ Sci B ; 8(2): 136-46, 2007 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-17266190

RESUMO

This study is aimed at investigating the potentials of ex vivo expansion and pluri-differentiation of cryopreservation of adult human bone marrow mesenchymal stem cells (hMSCs) into chondrocytes, adipocytes and neurocytes. Cryopreserved hMSCs were resuscitated and cultured for 15 passages, and then induced into chondrocytes, adipocytes and neurocytes with corresponding induction medium. The induced cells were observed for morphological properties and detected for expressions of type II collagen, triglyceride or neuron-specific enolase and nestin. The result showed that the resuscitated cells could differentiate into chondrocytes after exposure to transforming growth factor beta(1) (TGF-beta(1)), insulin-like growth factor I (IGF-I) and vitamin C (V(C)), and uniformly changed morphologically from a spindle-like fibroblastic appearance to a polygonal shape in three weeks. The induced cells were heterochromatic to safranin O and expressed cartilage matrix-procollagenal (II) mRNA. The resuscitated cells cultured in induction medium consisting of dexamethasone, 3-isobutyl-1-methylxanthine, indomethacin and IGF-I showed adipogenesis, and lipid vacuoles accumulation was detectable after 21 d. The resuscitated hMSCs were also induced into neurocytes and expressed nestin and neuron specific endolase (NSE) that were special surface markers associated with neural cells at different stage. This study suggested that the resuscitated hMSCs should be still a population of pluripotential cells and that it could be used for establishing an abundant hMSC reservoir for further experiment and treatment of various clinical diseases.


Assuntos
Células da Medula Óssea/citologia , Técnicas de Cultura de Células/métodos , Criopreservação/métodos , Células-Tronco Mesenquimais/citologia , Células-Tronco Pluripotentes/citologia , Engenharia Tecidual/métodos , Diferenciação Celular , Células Cultivadas , Humanos
11.
Zhonghua Wei Chang Wai Ke Za Zhi ; 9(6): 477-82, 2006 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-17143789

RESUMO

OBJECTIVE: To evaluate the clinical significance of detection on lymphatic microvessel, lymphatic microvessel density (LMVD) and vascular endothelial growth factor-C (VEGF-C) in patients with colorectal carcinoma. METHODS: Eighty tissue specimens of the colorectal carcinoma and the peritumoral tissue and thirty of adjacent normal bowel tissue were collected. The lymphatic microvessel and LMVD were determined by 5'-nucleotidase histochemical staining. The expression of VEGF-C protein and VEGF-C mRNA in specimens of colorectal carcinoma and normal colorectal tissues were studied by RT-PCR and immunohistochemical methods utilizing strept-avidin-biotin complex. Clinicopathological data and survival of each patient were obtained and analyzed. RESULTS: (1) The brown or filemot stained lymphatic microvessels were observed in specimens from the colorectal carcinoma, the peritumoral tissue and the normal bowel. Collapsed, nonfunctional lymphatic vessels were observed in the intratumoral tissue, and plenty of lymphatic vessels with large lumen referred as functional lymphatic vessels were observed in the peritumoral tissue. (2) The mean value of LMVD in the peritumoral tissue was significantly higher than that in the normal bowel tissue (9.76+/-2.85 vs. 5.49+/-1.43, t=8.220, P<0.01) and tumor tissue (9.76+/-2.85 vs. 2.13+/-0.96, t=15.118, P<0.001). (3) The positive rate (48.8% vs. 0, P<0.01) and mean value (1.09+/-1.20 vs. 0, P<0.01) of the VEGF-C protein expression in colorectal carcinoma specimens were significantly higher than that of the normal bowel tissue. The expression of VEGF-C protein was consistent with the expression of VEGF-C mRNA. The VEGF-C expression in intratumoral tissue demonstrated significant correlation with LMVD in the peritumoral tissue of colorectal carcinoma. (4) Both LMVD in the peritumoral tissue and the expression of VEGF-C in the intratumoral tissue correlated significantly with Dukes' stage (P<0.0001 and P=0.0234), lymph node metastasis (P<0.0001 and P=0.0059), and survival (P<0.0001 and P<0.0001), but not with age, sex, location and dimension of lesion, gross and histological type. Also, there was a positive significant correlation of LMVD in the peritumoral tissue with degree of differentiation (P=0.0168) and metastasis to the liver or the lung (P=0.0088). CONCLUSIONS: Lymphatic microvessels in the peritumoral tissue are functional. The functional lymphatic microvessels, increased LMVD in the peritumoral tissue and the expression of VEGF-C in the intratumoral tissue may act as the morphological features and the molecular phenotype of lymphangiogenesis in colorectal carcinoma, and also as important predictive markers for evaluating lymphatic metastasis and prognosis in patients with colorectal carcinoma.


Assuntos
Neoplasias Colorretais/metabolismo , Neoplasias Colorretais/patologia , Vasos Linfáticos/patologia , Fator C de Crescimento do Endotélio Vascular/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Linfangiogênese , Metástase Linfática , Masculino , Microvasos , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Prognóstico , Adulto Jovem
12.
Zhonghua Liu Xing Bing Xue Za Zhi ; 27(3): 200-3, 2006 Mar.
Artigo em Chinês | MEDLINE | ID: mdl-16792883

RESUMO

OBJECTIVE: To investigate the prevalence of domestic violence (DV) in Hunan. METHODS: Using a multi-stage sampling strategy, 9451 households involving 32 720 persons in urban, rural and industrial areas in Hunan, China were studied. Multiform clue investigation and face-to-face interviews were combined to investigate the prevalence of DV. RESULTS: A lifetime prevalence of DV was reported by 1533 households (16.2%). A total of 1098 households (11.6%) reported at least one incident of DV in the previous year. Both lifetime and 12-month prevalence of DV varied significantly by geographic setting (P < 0.01). The lifetime prevalence abuse rates were: spousal 10.2%, child abuse 7.8%, and elder 1.5%. With regard to household structure, the lifetime prevalence of DV was highest among those remarried families (21.0%), followed by married couples with one child and extended families with several generations living together (20.1% and 20.0%, respectively). The highest rate of spousal abuse was found among remarried families (14.7%), while child and elder abuse was most prevalent among extended families (12.4% and 4.1%, respectively). CONCLUSIONS: The findings suggested that although the prevalence of DV in Hunan was modest compared to Western countries, it remained a serious public health problem affecting over 1 in 10 households. Furthermore, the prevalence of various types of DV varied by geographic setting and family structure, suggesting that diverse geographic setting and family constellations carried different risk and protective features.


Assuntos
Maus-Tratos Infantis/estatística & dados numéricos , Abuso de Idosos/estatística & dados numéricos , Características da Família , Maus-Tratos Conjugais/estatística & dados numéricos , Idoso , Criança , China/epidemiologia , Estudos Epidemiológicos , Família , Feminino , Humanos , Masculino , Casamento/estatística & dados numéricos , Filho Único , Prevalência
13.
World J Gastroenterol ; 12(3): 393-402, 2006 Jan 21.
Artigo em Inglês | MEDLINE | ID: mdl-16489638

RESUMO

AIM: To establish a novel coculture system for ex vivo expansion of umbilical cord blood(UCB) hematopoietic progenitors using thrombopoietin (TPO)/Flt-3 ligand (FL)-transduced human marrow-derived mesenchymal stem cells (tfhMSCs) as feeder. METHODS: UCB CD34+ cells were isolated and cultured using four culture systems in serum-containing or serum-free medium. Suitable aliquots of cultured cells were used to monitor cell production, clonogenic activity, and long-term culture-initiating culture (LTC-IC) output. Finally, the severe-combined immunodeficient (SCID) mouse-repopulating cell (SRC) assay was performed to confirm ability of the cultured cells to reconstitute long-term hematopoiesis. RESULTS: There were no significant differences in the number of total nucleated cells among different culture systems in serum-containing medium during 21-d culture. However, on d 14, the outputs of CD34+ cells, CFU-C and CFU-GEMM in tfhMSCs coculture system were significantly enhanced. LTC-IC assay demonstrated that the tfhMSCs coculture system had the most powerful activity. The severe-combined immunodeficient (SCID) mouse repopulating cell (SRC) assay confirmed extensive ability of the expanded cells to reconstitute long-term hematopoiesis. Furthermore, PCR analysis demonstrated the presence of human hematopoietic cells in the bone marrow and peripheral blood cells of NOD/SCID mice. CONCLUSION: The TPO/FL-transduced hMSCs, in combination with additive cytokines, can effectively expand hematopoietic progenitors from UCB in vitro and the tfhMSCs coculture system may be a suitable system for ex vivo manipulation of primitive progenitor cells under contact culture conditions.


Assuntos
Antígenos CD34/metabolismo , Técnicas de Cultura de Células , Técnicas de Cocultura , Sangue Fetal/citologia , Células-Tronco Mesenquimais/metabolismo , Animais , Células Cultivadas , Ensaio de Unidades Formadoras de Colônias , Meios de Cultura Livres de Soro , Citocinas/metabolismo , Humanos , Imunofenotipagem , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Células-Tronco Mesenquimais/citologia , Camundongos , Camundongos SCID , Trombopoetina/genética , Trombopoetina/metabolismo
14.
Zhongguo Zhong Xi Yi Jie He Za Zhi ; 25(2): 154-9, 2005 Feb.
Artigo em Chinês | MEDLINE | ID: mdl-15768882

RESUMO

OBJECTIVE: To explore the molecular mechanism of wenban humai granule (WHG) in stabilizing atheromatous plaque, by observing its effect on the collagen degradation and synthesis imbalance manner in the fibrous cap of the plaque. METHODS: Atherosclerosis (AS) rabbit model established by feeding high fat diet. The changes of protein and mRNA expression of macrophage CD68, metalloproteinase-1 (MMP-1), alpha-smooth muscle actin (alpha-SMA) and collagen I (C-I) in model rabbits' neo-genesic intima were determined by immunohistochemical stain and in situ hybridization methods before and after treatment as well as before and after modeling. RESULTS: After being fed with high fat diet for 7 weeks, the protein and mRNA expression of macrophage CD68, MMP-1 in neo-genesic intima of aorta in the model rabbits significantly increased, these changes could be significantly restored after 8 weeks treatment with WHG or simvastatin. At the same time, the expressions of alpha-SMA protein and C-I protein and mRNA slightly increased due to the immigration of SMC in aortic media to neo-genesic intima, these expressions could be further increased after WHG treatment but showed a reducing trend after simvastatin treatment (P < 0.05 and P < 0.01). In the whole course, positive correlation was shown between protein expressions of CD68 and MMP-1 (r = 0.952, P < 0.01) and also between these of alpha-SMA and C-I (r = 0.793, P < 0.01). CONCLUSION: WHG affects the collagen degradation and synthesis imbalance in the fibrous cap of the plaque to stabilize plaque through bi-directional regulation, up-regulating synthesis thesis factors and down-regulating degradation factors, while simvastatin perform its action on plaque stability by down-regulating degradation factors alone.


Assuntos
Arteriosclerose/metabolismo , Arteriosclerose/patologia , Medicamentos de Ervas Chinesas/farmacologia , Actinas/metabolismo , Animais , Antígenos CD/metabolismo , Antígenos de Diferenciação Mielomonocítica/metabolismo , Aorta/patologia , Arteriosclerose/tratamento farmacológico , Colágeno/metabolismo , Medicamentos de Ervas Chinesas/uso terapêutico , Macrófagos/metabolismo , Metaloproteinase 1 da Matriz/metabolismo , RNA Mensageiro/metabolismo , Coelhos , Distribuição Aleatória
15.
Fish Shellfish Immunol ; 17(4): 353-66, 2004 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-15312662

RESUMO

The double-stranded-RNA-dependent protein kinase (PKR) is an important component in an antiviral defence pathway that is mediated by interferon (IFN) in vertebrates. Previously, some important IFN system genes had been identified from an IFN-producing CAB (crucian carp Carassius auratus blastulae embryonic) cells after treatment with UV-inactivated GCHV (grass carp haemorrhage virus). Here, a fish PKR-like gene, named CaPKR-like, is cloned and sequenced from the same virally infected CAB cells. It has 2192 base pairs in length with a largest open reading frame (ORF) encoding a protein of 513 amino acid residues. BLAST search reveals that the putative CaPKR-like protein is most homologous to human PKR and also has a high-level homology with all members of a family of eIF2alpha kinases. Structurally, CaPKR-like possesses a conserved C-terminal catalytic domain of eIF2alpha kinase family and the most similarity to mammalian PKRs. Within its N-terminus, there are no dsRNA-binding domains conserved in mammalian PKRs instead of two putative Z-DNA binding domains (Zalpha). Like mammalian PKRs, CaPKR-like had a very low level of constitutive expression in normal CAB cells but was up-regulated in response to active GCHV, UV-inactivated GCHV and CAB IFN, implying that the transcriptional activation of CaPKR-like by viral infection is mediated possibly by newly produced CAB IFN, which was further supported by using cycloheximide, a potent inhibitor of protein synthesis. The results together suggested that CaPKR-like was the first identified fish gene most similar to mammalian PKRs.


Assuntos
Carpas/genética , Filogenia , Reoviridae , eIF-2 Quinase/biossíntese , eIF-2 Quinase/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Blástula/citologia , Blástula/metabolismo , Células Cultivadas , Clonagem Molecular , Análise por Conglomerados , Cicloeximida , Primers do DNA , Indução Enzimática/efeitos dos fármacos , Indução Enzimática/fisiologia , Interferons/metabolismo , Interferons/farmacologia , Dados de Sequência Molecular , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Alinhamento de Sequência , Análise de Sequência de DNA , Raios Ultravioleta , Inativação de Vírus
16.
Fish Shellfish Immunol ; 15(5): 453-66, 2003 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-14550671

RESUMO

Interferon (IFN) can induce an antiviral state via interferon-regulatory transcription factors (IRFs), which bind to and control genes directed by the interferon-stimulated response element (ISRE). Here we describe a fish IRF, termed CaIRF7, cloned from a subtractive cDNA library which is constructed with mRNAs obtained from crucian carp (Carassius auratus L.) blastulae embryonic (CAB) cells infected by UV-inactivated GCHV and mock-infected cells. CaIRF7 cDNA was found to be 1816 bp in length, with a 42 bp 5'UTR and a 508 bp 3'UTR. The open reading frame translates into 421 amino acids in which a DNA-binding domain (DBD) containing the repeated tryptophan motif and IRFs association domain have been identified. Like chicken GgIRF3, CaIRF7 was most similar to mammalian IRF7 with 27 to 30% identity overall and some 37% identity in their DBDs. A single transcript of 1.9 kb was detected in virally induced CAB cells by virtual Northern blotting. RT-PCR analysis revealed a wide tissue distribution of CaIRF7 constitutive expression, with detectable transcript in non-infected CAB cells and various tissues of healthy crucian carp. In addition, CaIRF7 expression was differentially increased by stimulation of the CAB cells with active GCHV, UV-inactivated GCHV or CAB IFN, indicating that the activation of CaIRF7 was directly regulated by IFN.


Assuntos
Clonagem Molecular , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/imunologia , Carpa Dourada/imunologia , Animais , Sequência de Bases , Blástula/imunologia , Northern Blotting , Análise por Conglomerados , Primers do DNA , DNA Complementar/genética , Perfilação da Expressão Gênica , Carpa Dourada/genética , Fator Regulador 7 de Interferon , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Alinhamento de Sequência
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