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2.
Curr Med Sci ; 40(5): 817-821, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-33123896

RESUMO

In the period of regular epidemic prevention and control of Coronavirus disease 2019 (COVID-19) in our country, work resumption has been fully advanced. But there are still new sporadic local cases and imported cases across the country. In this situation, whether kindergartens reopening will increase the risk of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spread still remains uncertain. We reviewed two pediatric patients with moderate COVID-19, collected the epidemiologic information and monitored the cycle threshold value of rectal specimen and the viral loads, and discussed the transmission of SARS-CoV-2 in pediatric patients and the virulence of feces in children with moderate COVID-19, in order to analyze the risk of kindergartens reopening.


Assuntos
Betacoronavirus/isolamento & purificação , Infecções por Coronavirus/prevenção & controle , Fezes/virologia , Pandemias/prevenção & controle , Pneumonia Viral/prevenção & controle , Betacoronavirus/genética , Betacoronavirus/patogenicidade , COVID-19 , Pré-Escolar , China/epidemiologia , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/transmissão , Feminino , Humanos , Pandemias/estatística & dados numéricos , Pais , Pneumonia Viral/epidemiologia , Pneumonia Viral/transmissão , SARS-CoV-2 , Instituições Acadêmicas , Carga Viral , Eliminação de Partículas Virais
3.
World J Pediatr ; 16(3): 232-239, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32333248

RESUMO

In the early February, 2020, we called up an experts' committee with more than 30 Chinese experts from 11 national medical academic organizations to formulate the first edition of consensus statement on diagnosis, treatment and prevention of coronavirus disease 2019 (COVID-19) in children, which has been published in this journal. With accumulated experiences in the diagnosis and treatment of COVID-19 in children, we have updated the consensus statement and released the second edition recently. The current version in English is a condensed version of the second edition of consensus statement on diagnosis, treatment and prevention of COVID-19 in children. In the current version, diagnosis and treatement criteria have been optimized, and early identification of severe and critical cases is highlighted. The early warning indicators for severe pediatric cases have been summarized which is utmost important for clinical practice. This version of experts consensus will be valuable for better prevention, diagnosis and treatment of COVID-19 in children worldwide.


Assuntos
Infecções por Coronavirus , Coronavirus , Pandemias , Pneumonia Viral/epidemiologia , Betacoronavirus , COVID-19 , Criança , Consenso , Humanos , SARS-CoV-2
4.
Curr Med Sci ; 39(6): 899-905, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31845220

RESUMO

Absent in melanoma 2 (AIM2) inflammasome is a crucial link bridging the innate host defense and the subsequent adaptive immunity when activated by exogenous double stranded DNA (dsDNA). Through establishing models of disseminated murine cytomegalovirus (MCMV) infection in BALB/c and C57BL/6 mice, we evaluated dynamic expression of AIM2 inflammasome components and its relationship with pathological damage and viral replication, trying to figure out whether AIM2 inflammasome is related to the chronic mechanism of MCMV. BALB/c and C57BL/6 mice were sacrificed on day 0, 1, 3, 7, 14 and 28 post infection. Expression levels of AIM2, pro-caspase-1, caspase-1 p20, pro-IL1ß and mature IL1ß in primary peritoneal macrophages (PMs) and spleens were detected by Western blotting. Contents of IL18 in the serum were detected by ELISA. Pathological examinations of livers were performed, and mRNA levels of MCMV glycoprotein B (gB) in salivary glands also assessed. Results showed that expression levels of AIM2 in PMs and spleens of C57BL/6 mice increased on day 3, even continued to day 28; caspase-1 p20 and mature IL1ß increased on day 7, 14 and 28; the persistently high expression of IL18 in the serum started on day 1, showing a double peak curve. As for BALB/c mice, expression of AIM2 in PMs increased on day 1 and day 7, while contents of AIM2 in spleens increased on day 1 and day 3; caspase-1 p20 and mature IL1ß merely increased 7 days fter infection. Thereafter, expression levels of AIM2, caspase-1 p20, mature IL1ß and IL18 were limited; the duration of AIM2 inflammasome activation in BALB/c mice was much shorter than that in C57BL/6 mice. The severer pathological damage and more viral replications in BALB/c mice further proved the deficient antiviral immunity to MCMV. In conclusion, the activation of AIM2 inflammasome in BALB/c mice was short-lived, which is quite possibly related to the chronicity of MCMV infection.


Assuntos
Proteínas de Ligação a DNA/metabolismo , Infecções por Herpesviridae/imunologia , Inflamassomos/metabolismo , Muromegalovirus/patogenicidade , Animais , Caspase 1/metabolismo , Células Cultivadas , Modelos Animais de Doenças , Feminino , Regulação da Expressão Gênica , Interleucina-1beta/metabolismo , Macrófagos Peritoneais/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Muromegalovirus/genética , Muromegalovirus/imunologia , Baço/imunologia , Proteínas do Envelope Viral/genética
5.
Medicine (Baltimore) ; 98(39): e17305, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31574858

RESUMO

Until now, the recognition of sodium taurocholate cotransporting polypeptide (NTCP) deficiency has been mainly based on sporadic case reports. It was previously believed to be mildly symptomatic and resulting in mild liver dysfunction. However, to our knowledge, there have been no reports about the histopathologic and ultrastructural pathologic characteristics of the disease. The aim of the study was to analyze the clinical, histopathologic and ultrastructural pathologic characteristics of NTCP deficiency in 13 pediatric patients.From August 2012 to October 2018, this retrospective study conducted in the Department of Pediatrics of Tongji Hospital, China analyzed the data of 13 NTCP deficient patients with an SLC10A1 gene mutation. Except for NTCP deficiency, no other liver diseases were present in the patients, which was determined by both a genetic testing panel for jaundice and by reviewing medical records. The laboratory results, imaging, histopathologic, and ultrastructural pathologic information were recorded for analysis.The serum level of total bile acid was high in all 13 patients. All patients had adequate growth and development. Eight of the patients (8/13) presented with visible jaundice and 12 (12/13) were found to have hyperbilirubinemia. A needle liver biopsy was performed in 11 cases, which revealed slightly chronic inflammation in all 11 patients. One of the patients (1/13) was found to be suffering from gallstones.The data showed that although NTCP deficiency was often asymptomatic, some of the patients showed obvious clinical expressions, such as jaundice. Among the 13 pediatric patients with NTCP deficiency, both the biochemical and histopathologic features were similar to those of mild hepatocellular jaundice. In addition, it was determined that the clinical features in the patient with gallstones may have been caused by NTCP deficiency.


Assuntos
Ácidos e Sais Biliares/sangue , Icterícia , Hepatopatias , Fígado , Transportadores de Ânions Orgânicos Dependentes de Sódio , Simportadores , Desenvolvimento Infantil , Pré-Escolar , China/epidemiologia , Testes Genéticos/métodos , Humanos , Biópsia Guiada por Imagem/métodos , Lactente , Icterícia/diagnóstico , Icterícia/etiologia , Fígado/metabolismo , Fígado/patologia , Hepatopatias/diagnóstico , Hepatopatias/genética , Hepatopatias/fisiopatologia , Hepatopatias/terapia , Testes de Função Hepática/métodos , Glicoproteínas de Membrana/metabolismo , Mutação , Transportadores de Ânions Orgânicos Dependentes de Sódio/deficiência , Transportadores de Ânions Orgânicos Dependentes de Sódio/genética , Pediatria/métodos , Estudos Retrospectivos , Simportadores/deficiência , Simportadores/genética
6.
Curr Med Sci ; 38(4): 632-639, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30128872

RESUMO

Increasing evidence has revealed that maternal cytomegalovirus (CMV) infection may be associated with neurodevelopmental disorders in offspring. Potential relevance between the placental inflammation and CMV-related autism has been reported by clinical observation. Meanwhile, abnormal expression of Toll-like receptor 2 (TLR2) and TLR4 in placenta of patients with chorioamnionitis was observed in multiple studies. IL-6 and IL-10 are two important maternal inflammatory mediators involved in neurodevelopmental disorders. To investigate whether murine CMV (MCMV) infection causes alterations in placental IL-6/10 and TLR2/4 levels, we analyzed the dynamic changes in gene expression of TLR2/4 and IL-6/10 in placentas following acute MCMV infection. Mouse model of acute MCMV infection during pregnancy was created, and pre-pregnant MCMV infected, lipopolysaccharide (LPS)-treated and uninfected mice were used as controls. At E13.5, E14.5 and E18.5, placentas and fetal brains were harvested and mRNA expression levels of placental TLR2/4 and IL-6/10 were analyzed. The results showed that after acute MCMV infection, the expression levels of placental TLR2/4 and IL-6 were elevated at E13.5, accompanied by obvious placental inflammation and reduction of placenta and fetal brain weights. However, LPS 50 µg/kg could decrease the EL-6 expression at E13.5 and E14.5. This suggests that acute MCMV infection during pregnancy could up-regulate the gene expression of TLR2/4 in placental trophoblasts and activate them to produce more proinflammatory cytokine IL-6. High dose of LPS stimulation (50 µg/kg) during pregnancy can lead to down-regulation of IL-6 levels in the late stage. Imbalance of IL-6 expression in placenta might be associated with the neurodevelopmental disorders in progeny.


Assuntos
Infecções por Herpesviridae/metabolismo , Placenta/metabolismo , Complicações Infecciosas na Gravidez/metabolismo , Receptor 2 Toll-Like/metabolismo , Receptor 4 Toll-Like/metabolismo , Animais , Feminino , Infecções por Herpesviridae/genética , Interleucina-10/genética , Interleucina-10/metabolismo , Interleucina-6/genética , Interleucina-6/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Gravidez , Complicações Infecciosas na Gravidez/genética , Receptor 2 Toll-Like/genética , Receptor 4 Toll-Like/genética , Regulação para Cima
7.
World J Gastroenterol ; 23(48): 8570-8581, 2017 Dec 28.
Artigo em Inglês | MEDLINE | ID: mdl-29358865

RESUMO

AIM: To investigate the impact of fecal microbiota transplantation (FMT) treatment on allergic colitis (AC) and gut microbiota (GM). METHODS: We selected a total of 19 AC infants, who suffered from severe diarrhea/hematochezia, did not relieve completely after routine therapy or cannot adhere to the therapy, and were free from organ congenital malformations and other contraindications for FMT. Qualified donor-derived stools were collected and injected to the AC infants via a rectal tube. Clinical outcomes and follow-up observations were noted. Stools were collected from ten AC infants before and after FMT, and GM composition was assessed for infants and donors using 16S rDNA sequencing analysis. RESULTS: After FMT treatment, AC symptoms in 17 infants were relieved within 2 d, and no relapse was observed in the next 15 mo. Clinical improvement was also detected in the other two AC infants who were lost to follow-up. During follow-up, one AC infant suffered from mild eczema and recovered shortly after hormone therapy. Based on the 16S rDNA analysis in ten AC infants, most of them (n = 6) had greater GM diversity after FMT. As a result, Proteobacteria decreased (n = 6) and Firmicutes increased (n = 10) in post-FMT AC infants. Moreover, Firmicutes accounted for the greatest proportion of GM in the patients. At the genus level, Bacteroides (n = 6), Escherichia (n = 8), and Lactobacillus (n = 4) were enriched in some AC infants after FMT treatment, but the relative abundances of Clostridium (n = 5), Veillonella (n = 7), Streptococcus (n = 6), and Klebsiella (n = 8) decreased dramatically. CONCLUSION: FMT is a safe and effective method for treating pediatric patients with AC and restoring GM balance.


Assuntos
Colite/terapia , Transplante de Microbiota Fecal , Fezes/microbiologia , Microbioma Gastrointestinal/imunologia , Colite/imunologia , Colite/microbiologia , Diarreia/imunologia , Diarreia/microbiologia , Diarreia/terapia , Feminino , Seguimentos , Hemorragia Gastrointestinal/imunologia , Hemorragia Gastrointestinal/microbiologia , Hemorragia Gastrointestinal/terapia , Humanos , Lactente , Masculino , Recidiva , Resultado do Tratamento
8.
Zhonghua Er Ke Za Zhi ; 51(4): 260-4, 2013 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-23927798

RESUMO

OBJECTIVE: To investigate the genetic polymorphism of human cytomegalovirus (HCMV) glycoprotein H (gH) from infantile clinical isolates, to analyze the genotypic distribution of gH in different diseases of HCMV infection and try to find the correlations between the diseases and genotypes. METHOD: Fresh urine specimens were collected from the hospitalized children with different diseases whose blood HCMV-IgM and HCMV-IgG were positive. Virus was isolated from these specimens. Glycoprotein H of harvest clinical isolates was genotyped by nested-PCR combined with restriction fragment length polymorphism (RFLP), the purified PCR products were digested by restriction endonuclease HhaI. The digested products were genotyped by polyacrylamide gel electrophoresis and silver staining. Classification and results of sequencing were compared. RESULT: Totally 102 HCMV clinical isolates were obtained. Glycoprotein H gene of these clinical isolates (43 cases had infantile hepatitis syndrome, 38 cases had anicteric hepatitis, 13 pneumonia, 7 thrombocytopenic purpura, and 1 congenital CMV infection) were positive by nested-PCR, whose positive rate was 100%. The results showed that 62 strains were gH1 genotypes (60.8%), while 40 strains were gH2 (39.2%), mixed type or new genotype was not observed. In infantile hepatitis syndrome (26 clinical isolates were gH1 genotypes, 17 clinical isolates were gH2 genotypes), anicteric hepatitis (25 were gH1, 13 were gH2) and pneumonia (9 were gH1, 4 were gH2), the distribution of HCMV gH genotypes of infantile clinical isolates was consistent with the overall trend (χ(2) = 0.357, P > 0.05). However , the gH2 was more common than gH1 in the clinical isolates of patients with thrombocytopenic purpura (6 were gH2, 1 were gH2, χ(2) = 6.083, P < 0.05). CONCLUSION: Genotype 1 was the dominant genotype of glycoprotein H in HCMV clinical isolates from our hospital infants. There was no significant difference between the distribution of gH genotypes in infantile hepatitis syndrome, anicteric hepatitis and pneumonia. However, gH2 was the dominant genotype in thrombocytopenic purpura. These findings suggested that there may be a certain relevance between gH genotype and different clinical manifestations.


Assuntos
Infecções por Citomegalovirus/virologia , Citomegalovirus/classificação , Citomegalovirus/genética , Hepatite/virologia , Proteínas do Envelope Viral/genética , Sequência de Aminoácidos , Sequência de Bases , Pré-Escolar , Citomegalovirus/isolamento & purificação , Primers do DNA , DNA Viral/genética , Feminino , Genótipo , Humanos , Lactente , Recém-Nascido , Masculino , Pneumonia Viral/virologia , Reação em Cadeia da Polimerase , Polimorfismo Genético , Polimorfismo de Fragmento de Restrição , Urina/virologia
9.
Mol Med Rep ; 7(4): 1343-9, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23426791

RESUMO

Human cytomegalovirus (HCMV) has been associated with a wide spectrum of diseases. There is currently no effective treatment for eliminating the virus. Garlic bulb extract has been reported to possess anti-viral efficacy. This study aimed to investigate the expression of the immediate­early (IE; ul122 and ul123), early (E; ul54) and late (L; ul83) genes of HCMV as well as the inhibitory effect of allitridin on the transcription levels of these genes. The results indicated that a HCMV gene expression cascade occurred, and that the deletion of IE72 had no influence on the transcription of the ul122 gene, while it led to significant reductions of ul54 and ul83 mRNA expression levels. Additionally, allitridin effectively suppressed the transcription of the HCMV IE, E and L genes; the inhibition rates of the transcription of the ul122 and ul123 genes were higher compared with those of ul54 and ul83 mRNA expression, while the expression of the IE genes was not significantly reduced by ganciclovir (GCV). Our results indicate that the HCMV IE72 deletion mutant strain affects the transcription of the virus downstream gene, allitridin inhibits HCMV infection in vitro, and that the IE genes may be the key target of allitridin in its action against HCMV.


Assuntos
Compostos Alílicos/farmacologia , Citomegalovirus/efeitos dos fármacos , Sulfetos/farmacologia , Replicação Viral/efeitos dos fármacos , Compostos Alílicos/química , Citomegalovirus/genética , Citomegalovirus/patogenicidade , DNA Polimerase Dirigida por DNA/biossíntese , DNA Polimerase Dirigida por DNA/genética , Alho/química , Regulação Viral da Expressão Gênica/efeitos dos fármacos , Humanos , Proteínas Imediatamente Precoces/biossíntese , Proteínas Imediatamente Precoces/genética , Técnicas In Vitro , Fosfoproteínas/biossíntese , Fosfoproteínas/genética , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Sulfetos/química , Transativadores/biossíntese , Transativadores/genética , Proteínas da Matriz Viral/biossíntese , Proteínas da Matriz Viral/genética , Proteínas Virais/biossíntese , Proteínas Virais/genética
10.
J Med Virol ; 85(3): 493-500, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23341371

RESUMO

This study investigated the effects of allitridin compound on murine cytomegalovirus (MCMV)-induced regulatory T cell (Treg; CD4(+) CD25(+) Foxp3(+) ) amplification in vivo and in vitro. One hundred twenty MCMV-infected mice were allocated at random into two groups for treatment with allitridin or placebo. Another 120 mock-infected mice were randomly allocated as controls for the allitridin treatment and placebo treatment groups. The mice were euthanized at various time points after infection (out to 120 days) to evaluate the effects of treatment on Treg presence and function, as well as MCMV infective load. Co-culture with mouse embryo fibroblasts (MEF) and MCMV was performed to evaluate allitridin-mediated Treg and anti-CMV effects. The maximum tolerance concentration (MTC) of allitridin was used to treat cells for 3 days. Changes in Foxp3 mRNA and protein levels, percentages of T cell subsets, and Treg-related cytokines (IL-10 and TGF-ß) were measured. Allitridin treatment did not influence Foxp3 expression and Treg proportion in uninfected mice, but did down-regulate each in infected mice during the chronic infection period. Additionally, allitridin treatment reduced the MCMV load in salivary glands. MTC allitridin treatment of co-cultures partially blocked MCMV induction of Foxp3 mRNA and protein expression. In vitro treatment with allitridin also increased significantly the percentages of Tc1, Tc2, and Th1, reduced the secreted levels of IL-10 and TGF-ß1, and significantly suppressed viral loads. In conclusion, allitridin can promote MCMV-induced Treg expansion and Treg-mediated anti-MCMV immunosuppression. Therefore, allitridin may be useful as a therapeutic agent to enhance the specific cellular immune responses against CMV.


Assuntos
Compostos Alílicos/administração & dosagem , Infecções por Herpesviridae/imunologia , Fatores Imunológicos/administração & dosagem , Muromegalovirus/imunologia , Sulfetos/administração & dosagem , Linfócitos T Reguladores/efeitos dos fármacos , Compostos Alílicos/isolamento & purificação , Animais , Citocinas/metabolismo , Modelos Animais de Doenças , Feminino , Fatores de Transcrição Forkhead/biossíntese , Alho/química , Perfilação da Expressão Gênica , Fatores Imunológicos/isolamento & purificação , Camundongos , Camundongos Endogâmicos BALB C , Placebos/administração & dosagem , Sulfetos/isolamento & purificação , Subpopulações de Linfócitos T/imunologia , Carga Viral
12.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 28(6): 564-7, 2012 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-22691343

RESUMO

AIM: To explore the immunopathological mechanism of spleen damage in acute disseminated infection of murine cytomegalovirus (MCMV) in vivo by observing the association of virus titers and the expressions of caspase-1 and pro-inflammatory factors (IL-1ß and IL-18) with the degree of pathological damage of the spleen. METHODS: BALB/c mice (n=24) were randomly divided into 2 groups (n=12 per group), experimental group and control group. The control mice were sham-infected. The experimental mice were infected with MCMV Smith for establishing the models of acute disseminated MCMV infection. Three mice of each group were randomly chosen to be killed on day 3, 7, 14 and 28 after infection, respectively. Viral titers in the spleen tissues were determined using a standard plaque assay; the expression of caspase-1 in the splenocytes was detected by Western blot; the expressions of IL-1ß and IL-18 in the spleen tissues were observed by immunohistochemical staining; the degree of spleen histological damage was observed by HE staining and graded by a semiquantitative method. RESULTS: Viral titers in the spleen peaked on day 3, but quickly diminished on day 7 and virus was not detected in the spleen on day 14 after infection. Compared with the normal control group, the protein levels of caspase-1 in MCMV-infected mice were markedly elevated on day 3 (P<0.01), and then dropped slowly; the expressions of IL-1ß and IL-18 in the spleen tissues gradually increased to the climax on day 7, then decreased on day 14 and turned to the normal on day 28. However, the pathological condition of the spleen in infected mice deteriorated gradually until day 14, and then showed obviously the signs of recovery on day 28. The spleen damage was aggravated following the elevation of IL-1ß and IL-18 expressions and alleviated after they declined. CONCLUSION: MCMV infection would stimulate the increase of caspase-1 expression and the production of pro-inflammatory factors (IL-1ß and IL-18). IL-1ß and IL-18 not only exert antiviral effect, but also might be involved in the immunopathological injury of spleen tissue during the acute disseminated MCMV infection.


Assuntos
Infecções por Citomegalovirus/imunologia , Infecções por Citomegalovirus/patologia , Muromegalovirus/imunologia , Baço/imunologia , Baço/patologia , Animais , Caspase 1/metabolismo , Infecções por Citomegalovirus/virologia , Interleucina-18/metabolismo , Interleucina-1beta/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Baço/virologia , Carga Viral
13.
Arch Virol ; 156(10): 1841-6, 2011 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-21604182

RESUMO

This study investigated the effects of allitridin on acute and chronic mouse cytomegalovirus (MCMV) infections in vivo. The results demonstrated that allitridin reduced the titers of MCMV in salivary glands, and reductions in viral loads were confirmed by determining viral DNA and RNA levels in susceptible organs during the acute infection phase. Although allitridin did not eliminate MCMV, treatment reduced viral levels and facilitated healing of pathologic lesions in organs, particularly during the chronic infection phase. The results presented in this report suggest that allitridin could act as an effective agent against MCMV infections in vivo.


Assuntos
Compostos Alílicos/farmacologia , Antivirais/farmacologia , Infecções por Citomegalovirus/virologia , Infecções por Herpesviridae/virologia , Muromegalovirus/efeitos dos fármacos , Sulfetos/farmacologia , Doença Aguda/terapia , Animais , Doença Crônica/terapia , Citomegalovirus/efeitos dos fármacos , Citomegalovirus/fisiologia , Infecções por Citomegalovirus/tratamento farmacológico , Modelos Animais de Doenças , Feminino , Infecções por Herpesviridae/tratamento farmacológico , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Muromegalovirus/fisiologia , Glândulas Salivares/virologia , Carga Viral/efeitos dos fármacos
14.
Zhonghua Er Ke Za Zhi ; 49(10): 788-92, 2011 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-22321188

RESUMO

OBJECTIVES: To analyze the clinical features of acute invasive pulmonary aspergillosis in younger children, in order to improve the levels of early recognition, diagnosis and management of this disease. METHOD: Clinical data of 8 patients aged below 15 months who were diagnosed as acute invasive pulmonary aspergillosis from August 2010 to February 2011 in general pediatric wards in our hospital were retrospectively analyzed for the high-risk factors of the hosts, clinical manifestations, laboratory findings and lung CT imaging, the processes of diagnosis and treatment, and the outcomes. RESULT: Five cases were tested for serum GM test absorbent index (GMI) ranged from 1.92 to 3.27; in 2 cases sputum culture was positive for Aspergillus fumigatus for twice, and 1 infant was serum GMI 2.85 and a sputum culture was positive for Aspergillus fumigatus positive, all these findings were accordant with the clinical diagnosis. Seven cases had a history of receiving intravenously broad-spectrum antibiotics or plus corticosteroids (6 hospitalized, 1 out-patient), and one was only 1 month old, whose parents had severe tinea pedis. 4 patients of high-fever type had sustained high temperature, severe changes of lungs without obvious respiratory symptoms and signs in early phase, and significant increase of the rod granulocyte rate (0.25 - 0.68), which was apparently discordant with the normal WBC count and high sensitivity C-reactive protein (hs-CRP) value. Another 4 cases of non-high-fever type were present with normal WBC count, hs-CRP value and the percentage of rod granulocyte. Among them, 3 infants had low-grade fever, with serious respiratory symptoms and signs and changes of lungs CT. Another 1-month-old case only showed lower vigor and response. Lung CT imaging often showed multiple irregular large nodules, patches and streaks of density (6 cases) and unilateral lobar consolidation (1 case), with some involving the pleura; one appeared severe peri-main bronchus lesions with stenoses of bilateral main bronchi. The first case died of multiple organ failure because of severe sepsis complication. Another 7 cases were treated with voriconazole promptly after clinical or suspected diagnosis, and the state of patients relieved rapidly within 1 - 3 d. CONCLUSION: The abuse of broad-spectrum antibiotics and corticosteroids may increase the risk of invasive pulmonary aspergillosis in younger children. There may be the risk of nosocomial infection and spread of aspergillus in general pediatric wards. Cases of high-fever type in early period of disease had two inconsistency: few symptoms and signs, while severe changes of lungs CT; apparent increase of peripheral rod granulocyte, while normal WBC count and hs-CRP value. Preemptive voriconazole therapy could obtain significant effect and reduce the mortality rate.


Assuntos
Aspergilose Pulmonar Invasiva , Doença Aguda , Corticosteroides/efeitos adversos , Antibacterianos/efeitos adversos , Aspergillus fumigatus/isolamento & purificação , Feminino , Humanos , Lactente , Aspergilose Pulmonar Invasiva/diagnóstico , Aspergilose Pulmonar Invasiva/terapia , Masculino , Estudos Retrospectivos , Fatores de Risco
15.
Chin Med J (Engl) ; 124(21): 3532-6, 2011 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-22340173

RESUMO

BACKGROUND: Murine cytomegalovirus (MCMV) early protein M112-113 is involved in viral DNA replication and believed to play a crucial role in the viral pathogenesis. To investigate the biological function of M112-113 protein in the pathogenesis of the brain disorders caused by cytomegalovirus (CMV), a screening for proteins interacting with M112-113 was performed by a yeast two-hybrid system. METHODS: Bait plasmid pGBKT7-M112-113 was constructed and transformed into AH109 yeast. After confirmation of the expression of MCMV M112-113 in yeast, the bait yeast was mated with a prey yeast containing mouse brain cDNA library plasmid to screen the proteins interacting with M112-113. Interactions between M112-113 and the obtained proteins were verified by yeast two-hybrid assay and chemiluminescent co-immunoprecipitaion. RESULTS: Two proteins interacting with M112-113 were identified, including metastasis-associated 1 (MTA1) and zinc finger, CCHC domain containing 18 (ZCCHC18). M112-113 protein could interact with MTA1 or ZCCHC18 in yeast and mammalian cells. CONCLUSION: The interactions of M112-113 with MTA1 or ZCCHC18 may be related to the pathogenesis of MCMV-associated disease in central nervous system.


Assuntos
Encéfalo/metabolismo , Muromegalovirus/metabolismo , Proteínas Virais/metabolismo , Animais , Linhagem Celular , Humanos , Imunoprecipitação , Camundongos , Plasmídeos , Ligação Proteica , Técnicas do Sistema de Duplo-Híbrido
17.
Zhonghua Yi Xue Za Zhi ; 88(42): 2999-3002, 2008 Nov 18.
Artigo em Chinês | MEDLINE | ID: mdl-19080080

RESUMO

OBJECTIVE: To explore the effects of acute and chronic murine cytomegalovirus (MCMV) infections on the regulatory T cells (Treg) ratio and protein expression of the Th1/Th2 transcription factors T-bet/GATA-3. METHODS: 120 BALB/c mice were randomly divided into 2 equal groups: MCMV-infected group undergoing infra-peritoneal injection of homogenate of salivary gland containing MCMV, and mock infection group undergoing infra-peritoneal injection of normal homogenate of salivary gland 1, 3, 7, 14, 28, 45, 60, 75, 90, and 120 days after infection 6 mice from each group were killed to examine the viral load of the heart, lung, liver, and kidney by plaque assay to access the status of MCMV infection. Suspension of splenocytes was prepared. The proportion of CD4+CD25+Foxp3+Treg in the splenocytes was measured by flow cytometry. Western blotting was used to detect the protein expression of T-bet/GATA-3. RESULTS: The cutoff point between acute and chronic points was the 28th day. The CD4+CD25+Foxp3+Treg proportion in splenocytes significantly decreased during the acute infection stage and to the lowest level of (1.46+/-0.27)% at day 28, significantly lower than that of the mock infection group [(2.78+/-0.29)%, P<0.05]; then obviously increased in the chronic infection stage, increased to (4.51+/-0.24)% at day 60, significantly higher than that of the mock infection group [(2.69+/-0.12)%, P<0.05], and continued to increase still. The protein level (K value) of T-bet of the MCMV infection group peaked to the level of (0.618+/-0.053) on day 3, obviously higher than that of the mock infected group [(0.205+/-0.026)], then decreased to the level similar to that of the mock infection group on day 28, and was obviously lower than that of the mock infection group on day 75. Whereas the protein level of GATA-3 of the MCMV group increased to (0.836+/-0.061) on day 3, markedly higher than that of the mock infection group (0.398+/-0.022), peaked on day 7, then gradually decreased, and remained at the levels similar to those of the mock infection group from day 75 to day 120. CONCLUSION: In the acute infection stage, MCMV up-regulates the T-bet and GATA-3 protein expression. But during the chronic infection stage, MCMV induces a marked proliferation and activation of Treg cells which further inhibit the Th1 and Th2 reactions, especially Th1 response. Treg proliferation may be an important mechanism of chronic and persistent CMV infection in the host.


Assuntos
Fator de Transcrição GATA3/metabolismo , Infecções por Herpesviridae/metabolismo , Proteínas com Domínio T/metabolismo , Linfócitos T Reguladores/metabolismo , Doença Aguda , Animais , Diferenciação Celular , Doença Crônica , Modelos Animais de Doenças , Feminino , Citometria de Fluxo , Infecções por Herpesviridae/genética , Camundongos , Camundongos Endogâmicos BALB C , Muromegalovirus/patogenicidade , Baço/citologia , Linfócitos T Reguladores/citologia , Células Th1/metabolismo , Células Th2/metabolismo
18.
Antiviral Res ; 72(1): 68-74, 2006 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-16844239

RESUMO

Garlic (Allium sativum) extraction has been reported having anti-HCMV efficacy. This study was aimed to investigate the effect of allitridin (diallyl trisulfide, a compound from A. sativum extraction) on the replication of HCMV and the expression of viral immediate-early genes. In HCMV plaque-reduction assay, allitridin appeared a dose-dependent inhibitory ability with EC(50) value of 4.2 microg/ml (selective index, SI=16.7). Time-of-addition and time-of-removal studies showed that allitridin inhibited HCMV replication in earlier period of viral cycle before viral DNA synthesis. Both immediate early gene (ie1) transcription and IEA (IE(1)72 and IE(2)86) expression was suppressed by allitridin, but not by GCV in a single HCMV cycle format. In addition, allitridin appeared stronger inhibition on IE(2)86 than on IE(1)72. Decrease of viral DNA load in infected cells was also detected under allitridin treatment, probably due to an indirect consequence of the reduction in ie gene transcription. In summary, this study indicated that allitridin has anti-HCMV activity and the mechanism is associated with suppression of ie gene transcription.


Assuntos
Compostos Alílicos/farmacologia , Antivirais/farmacologia , Citomegalovirus/efeitos dos fármacos , Sulfetos/farmacologia , Linhagem Celular , Citomegalovirus/genética , Citomegalovirus/fisiologia , Relação Dose-Resposta a Droga , Expressão Gênica/efeitos dos fármacos , Genes Precoces/efeitos dos fármacos , Genes Virais/efeitos dos fármacos , Humanos , Proteínas Imediatamente Precoces/genética , Transativadores/genética , Transcrição Genética/efeitos dos fármacos , Proteínas Virais/genética , Replicação Viral/efeitos dos fármacos
19.
Chin Med J (Engl) ; 118(23): 1994-9, 2005 Dec 05.
Artigo em Inglês | MEDLINE | ID: mdl-16336836

RESUMO

BACKGROUND: The production of neural stem cells (NSCs) derived from embryonic stem (ES) cells was usually very low according to previous studies, which was a major obstacle for meeting the needs of clinical application. This study aimed at investigating whether astrocytes could promote production of NSCs derived from ES cells in vitro. METHODS: Mouse ES cells line-D3 was used to differentiate into NSCs with astrocytes as inducing stromal cells by means of three-stage differentiation procedure. Another group without astrocytes served as control. The totipotency of ES cells was identified by observation of cells' morphology and formation of teratoma in severe combined immunodeficiency disease (SCID) mice. The quantity and purity of NSCs derived from ES cells were analyzed using clonogenic assay, immunohistochemical staining and flow cytometry assay. The plasticity of NSCs was detected by differentiating test. Octamer-binding transcription factor 4 (Oct-4) and nestin, the specific marker genes of ES cells and NSCs respectively, were detected continuously using reverse transcription-polymerase chain reaction (RT-PCR) method to monitor the process of cell differentiation. RESULTS: The ES cells of D3 line could maintain the ability of differentiating into cellular derivations of all three primary germ layers after continuous passage culture. At the end of two-stage of inducing process, 23.2 +/- 3.5 neurospheres per plate formed in astrocyte-induced group and only 0.8 +/- 0.3 per plate in the control group (clonogenic assay, P < 0.01), and the ratio of nestin positive cells was (50.2 +/- 2.8)% in astrocyte-induced group and only (1.4 +/- 0.5)% in the control group (flow cytometry, P < 0.01). With the induction undergoing, the expression of Oct-4 gradually decreased and then disappeared, while the expression of nestin was increased step by step, and the ratio of nestin positive cells was up to 91.4% by the three-stage differentiation. The nestin positive cells could be further induced into neurons, astrocytes, and oligodendrocytes in differentiating medium supplemented with fetal calf serum. The results of differentiating test showed that the ratio of NF-200 and NSE positive cells was (42.7 +/- 2.6)% in astrocyte-induced group and only (11.2 +/- 1.8)% in the control group (P < 0.01). CONCLUSIONS: Astrocytes can not only increase the production of NSCs derived from ES cells but also promote the differentiation of NSCs toward neuronal lineage.


Assuntos
Astrócitos/fisiologia , Diferenciação Celular , Embrião de Mamíferos/citologia , Neurônios/citologia , Células-Tronco/citologia , Animais , Linhagem da Célula , Células Cultivadas , Camundongos
20.
World J Gastroenterol ; 10(19): 2818-22, 2004 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-15334677

RESUMO

AIM: To investigate the different effects of mesenchymal stem cells (MSCs) and hematopoietic stem cells (HSCs) on hepatic differentiation. METHODS: MSCs from rat bone marrow were isolated and cultured by standard methods. HSCs from rat bone marrow were isolated and purified by magnetic activated cell sorting. Both cell subsets were induced. Morphology, RT-PCR and immunocytochemistry were used to identify the hepatic differentiation grade. RESULTS: MSCs exhibited round in shape after differentiation, instead of fibroblast-like morphology before differentiation. Albumin mRNA and protein were expressed positively in MSCs, without detection of alpha-fetoprotein (AFP). HSCs were polygonal in shape after differentiation. The expression of albumin signal decreased and AFP signal increased. The expression of CK18 was continuous in MSCs and HSCs both before and after induction. CONCLUSION: Both MSCs and HSCs have hepatic differentiation capabilities. However, their capabilities are not the same. MSCs can differentiate into mature hepatocyte-like cells, never expressing early hepatic specific genes, while Thy-1.1(+) cells are inclined to differentiate into hepatic stem cell-like cells, with an increasing AFP expression and a decreasing albumin signal. CK18 mRNA is positive in Thy-1.1(+) cells and MSCs, negative in Thy-1.1(-) cells. It seems that CK18 has some relationship with Thy-1.1 antigen, and CK18 may be a predictive marker of hepatic differentiation capability.


Assuntos
Diferenciação Celular/fisiologia , Células-Tronco Hematopoéticas/citologia , Mesoderma/citologia , Células-Tronco/citologia , Animais , Sequência de Bases , Biomarcadores/análise , Células da Medula Óssea/citologia , Primers do DNA , Citometria de Fluxo , Separação Imunomagnética , Masculino , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , alfa-Fetoproteínas/análise
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