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1.
Int J Oral Maxillofac Surg ; 52(8): 839-846, 2023 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-36653273

RESUMO

A protuberant shape and sufficient volume are the most important parameters for total tongue reconstruction. The conventional pectoralis major myocutaneous (PMMC) flap undergoes collapse due to atrophy of the denervated muscle. In a new technique, this flap was rolled up like sushi to reshape the neotongue. This study explored the feasibility and effect of the 'sushi roll' technique for precise total functional reconstruction of the tongue using a PMMC flap. Thirty patients scheduled for total glossectomy and PMMC flap reconstruction were recruited. The sushi roll technique was performed in 15 patients and the conventional repair in 15 patients. Outcomes were compared between the two groups. The flap survived in all 30 patients. The sushi roll group showed superior results to the conventional group in terms of time to oral alimentation (P = 0.012) and decannulation (P = 0.041), as well as swallowing function (P = 0.032), speech intelligibility (P < 0.001), shape (P < 0.001), and quality of life score (P < 0.001) at 12 months. The innovative sushi roll technique uses a folding method that utilizes the length rather than the thickness and width of the flap to maintain the volume and protuberance of the neotongue, which results in acceptable function and improved quality of life.


Assuntos
Retalho Miocutâneo , Neoplasias da Língua , Humanos , Músculos Peitorais/transplante , Qualidade de Vida , Neoplasias da Língua/cirurgia , Língua/cirurgia , Glossectomia/métodos
2.
Zhonghua Yu Fang Yi Xue Za Zhi ; 56(4): 427-432, 2022 Apr 06.
Artigo em Chinês | MEDLINE | ID: mdl-35488538

RESUMO

Objective: To identify and analyze two strains of C. diphtheriae in Guangdong Province by combining whole genome sequencing with traditional detection methods. Methods: The C. diphtheriae was isolated from Guangzhou in 2010 and Zhuhai in 2020 respectively. Isolates were identified by API Coryne strips and MALDI-TOF-MS. Genomic DNA was sequenced by using Illumina. The assembly was performed for each strain using CLC software. J Species WS online tool was used for average nucleoside homology identification, then narKGHIJ and tox gene were detected by NCBI online analysis tool BLSATN. MEGA-X was used to build a wgSNP phylogenetic tree. Results: GD-Guangzhou-2010 was Belfanti and GD-Zuhai-2020 was Gravis. ANIb between GD-Guangzhou-2010 and C. belfantii was 99.61%. ANI between GD-Zhuhai-2020 and C. diphtheriae was 97.64%. BLASTN results showed that the nitrate reduction gene narKGHIJ and tox gene of GD-Guangzhou-2010 was negative, while GD-Zhuhai-2020 nitrate reduction gene narKGHIJ was positive. There were two obvious clades in wgSNP phylogenetic tree. The first clades included all Mitis and Gravis types strains as well as GD-Zhuhai-2020. The second clades contained all isolates of C.belfantii, C.diphtheriae subsp. lausannense and GD-guangzhou-2010. Conclusion: Two non-toxic C. diphtheriae strains are successfully isolated and identified. The phylogenetic tree suggests that GD-Guangzhou-2010 and GD-Zhuhai-2020 are located in two different evolutionary branches.


Assuntos
Corynebacterium diphtheriae , Difteria , China/epidemiologia , Corynebacterium , Corynebacterium diphtheriae/genética , Difteria/epidemiologia , Difteria/microbiologia , Humanos , Nitratos , Filogenia
3.
Zhonghua Gan Zang Bing Za Zhi ; 29(11): 1124-1127, 2021 Nov 20.
Artigo em Chinês | MEDLINE | ID: mdl-34933438

RESUMO

Histones are important structural proteins of chromatin in the nucleus, which can regulate gene transcription, and can be released from the nucleus to the outside of the cell under injury and inflammatory stimulations, thereby causing cytotoxicity and immune stimulation, and aggravating tissue damage. Extracellular histones are involved in the occurrence and development of many diseases, including sepsis, autoimmune diseases, liver injury, and acute lung injury. Therefore, its application not only can be used as a body's biomarker of inflammation, but also it is expected to become a molecular target for the treatment of diseases. This article reviews the role of extracellular histones in the inflammatory process of liver injury.


Assuntos
Doenças Autoimunes , Histonas , Humanos , Inflamação , Fígado
4.
Zhonghua Kou Qiang Yi Xue Za Zhi ; 56(11): 1074-1079, 2021 Nov 09.
Artigo em Chinês | MEDLINE | ID: mdl-34763401

RESUMO

Objective: To observe the plaque accumulation at the fitting surface and oral hygiene status in patients with full-arch implant-supported fixed prostheses, and explore the possible influencing factors. Methods: Twenty-eight patients [17 males and 11 females, (63.0± 10.8) years old] with 40 full-arch implant-supported fixed prostheses (18 maxillary and 22 mandibular) were collected from January 2012 to September 2020 in Department of Implantology, Peking University School and Hospital of Stomatology. Plaque accumulation at the fitting surfaces were evaluated during the follow-up visit after 6 months following definitive prostheses delivery, by analyzing the digital photographs recorded by ImageJ. Meanwhile, the cleanliness of the fitting surface of prostheses and oral hygiene status were recorded. The oral hygiene habits and the patients' satisfaction with the prostheses were investigated by questionnaire, and the difference of plaque accumulation between different cleaning habits of dentures were compared. Results: The debris index of the fitting surface of the 40 full-arch implant-supported fixed prostheses were 3.28±0.75, and the percentage of area covered with plaque was (51.6±19.0)%. The debris index and plaque accumulation of the mandibular prostheses were significantly higher than that of the maxillary prostheses (P<0.05). In most mandibular prostheses (16/22), calculus was attached to the lingual side of the anterior tooth area. The reserved cleaning space of the restoration showed more plaque accumulation than in other parts. There was no significant difference in the percentage of area covered with plaque between groups with different cleaning habits. The satisfaction survey results indicated the "clean" project had a lowest score. Conclusions: The cleanliness of patients with full-arch implant-supported fixed prostheses was poor, and the hygiene status of the mandibular prostheses was worse than that of maxillary, especially in the anterior tooth area of mandibular prostheses. The influence of different cleaning habits on plaque accumulation was not observed.


Assuntos
Implantes Dentários , Placa Dentária , Idoso , Estudos Transversais , Prótese Dentária Fixada por Implante , Feminino , Seguimentos , Humanos , Masculino , Mandíbula , Pessoa de Meia-Idade
5.
Zhonghua Gan Zang Bing Za Zhi ; 27(5): 397-400, 2019 May 20.
Artigo em Chinês | MEDLINE | ID: mdl-31177669

RESUMO

Liver failure is a familiar severe disease, with no good clinical early diagnostic indicators and treatment methods. Studies have shown that non-encoding RNA (ncRNA) characterized by microRNA (miRNA) and long non-coding RNA (lncRNA) can be used not only as an early diagnostic indicator of liver failure, but also play a key regulatory role in an inflammatory response to liver failure, hepatocyte death and hepatocyte regeneration. Simultaneously, the epigenetic regulation of ncRNA also participates in the initiation and progression of liver failure. This article reviews the relationship between miRNA, lncRNA, and liver failure to find new targets for the diagnosis and treatment of liver failure.


Assuntos
Falência Hepática/genética , MicroRNAs , RNA Longo não Codificante , Epigênese Genética , Humanos , RNA não Traduzido
6.
J Viral Hepat ; 25(9): 1017-1025, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-29624802

RESUMO

The kinetics of serum hepatitis B surface antigen (HBsAg) during the natural history of hepatitis B virus (HBV) infection has been studied, but the factors affecting them remain unclear. We aimed to investigate the factors affecting HBsAg titres, using data from multicentre, large-sized clinical trials in China. The baseline data of 1795 patients in 3 multicentre trials were studied, and the patients were classified into 3 groups: hepatitis B early antigen (HBeAg)-positive chronic HBV infection (n = 588), HBeAg-positive chronic hepatitis B (n = 596), and HBeAg-negative chronic hepatitis B (n = 611). HBsAg titres in the different phases were compared, and multiple linear progression analyses were performed to investigate the implicated factors. HBsAg titres varied significantly in different phases (P = .000), with the highest (4.60 log10 IU/mL [10%-90% confidence interval: 3.52 log10 IU/mL-4.99 log10 IU/mL]) in patients with HBeAg-positive chronic HBV infection. In all phases, age and HBV DNA were correlated with serum HBsAg level. In HBeAg-positive chronic hepatitis B patients, a negative correlation between HBsAg titres and fibrosis stage was observed. Alanine amonitransferase or necroinflammatory activity was also correlated with HBsAg titres in HBeAg-negative chronic hepatitis B patients. In conclusion, decreased HBsAg titres may be associated with advancing fibrosis in HBeAg-positive chronic hepatitis B patients or increased necroinflammation in those with HBeAg-negative chronic hepatitis B. Our findings may help clinicians better understand the kinetics of HBsAg and provide useful insights into the management of this disease.


Assuntos
Antígenos de Superfície da Hepatite B/sangue , Hepatite B Crônica/complicações , Hepatite B Crônica/patologia , Cirrose Hepática/patologia , Soro/química , Adulto , Alanina Transaminase/sangue , China , DNA Viral/sangue , Feminino , Antígenos E da Hepatite B/sangue , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem
7.
Zhonghua Gan Zang Bing Za Zhi ; 25(9): 695-700, 2017 Sep 20.
Artigo em Chinês | MEDLINE | ID: mdl-29108192

RESUMO

Objective: To investigate the protective effect of ACY1215 (Rocilinostat), a histone deacetylase inhibitor, against brain edema in mice with acute liver failure. Methods: Lipopolysaccharide combined with D-galactosamine was used to establish a mouse model of acute liver failure, and ACY1215 was used for intervention. The effect of ACY1215 on histopathological changes of the liver was observed after 24 hours, as well as the changes in alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood ammonia, tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), brain water content, blood-brain barrier structure, NF-κB-p65, histone, acetylated histone, and TNF-α mRNA in brain tissue. Results: The mice with acute liver failure had marked pathological damage in liver tissue, as well as significant increases in the levels of ALT, AST, blood ammonia, TNF-α, and IFN-γ (t≥5.367, all P < 0.05). ACY1215 significantly improved the pathological damage in liver tissue and reduced the serum levels of ALT, AST, blood ammonia, TNF-α, and IFN-γ (t≤-3.515, all P < 0.05). ACY1215 also significantly reduced the expression of NF-κB-p65 (t = -5.871, P = 0.004) and the mRNA expression of TNF-α (t = -11.913, P < 0.01) in brain tissue and brain water content (t = -2.355, P < 0.01). According to the results of electron microscopy, the model group had an abnormal blood-brain barrier structure, and the ACY1215 group had slighter damage than the model group. Compared with the normal group, the model group had significant increases in the acetylation level of histone H3 and H4 in brain tissue (t≥3.009, both P < 0.05), while ACY1215 further upregulated the acetylation levels of histone H3 and H4 (t≥6.682, both P < 0.05). Conclusion: ACY1215 exerts a protective effect against brain edema in mice with acute liver failure, possibly by regulating histone acetylation and inhibiting inflammation.


Assuntos
Edema Encefálico , Inibidores de Histona Desacetilases/farmacologia , Animais , Aspartato Aminotransferases/sangue , Galactosamina , Fígado/patologia , Falência Hepática Aguda , Camundongos , NF-kappa B , Fator de Necrose Tumoral alfa/sangue
8.
Artigo em Chinês | MEDLINE | ID: mdl-29081098

RESUMO

Objective: To observe the therapeutic efficacy of alanyl glutamine injection on patients with gastrointestinal function obstacle caused by severe phorate poisoning. Methods: A total of 80 eligible patients with gastrointestinal function obstacle caused by severe phorate poisoning were randomly divided into the control group (n=40) and treatment group (n=40) . The control group was treated with the conventional therapy, which included forbidden diet, atropine, pralidoxime iodide, anti-inflammatory, albumin infusion, ω-3 fish oil fat emulsion, protection of organs function, blood perfusion, and Fat Emulsion, Amino Acids (17) and Glucose Injection. The treatment group was treated with alanyl glutamine injection plus the conventional therapy. To observe the time of recovering to normal of gastrointestinal function between the two groups, compared the AChE activity and changes of prealbumin, albumin and total protein of the two groups respectively. Furthermore, the total atropine dosage, the total pralidoxime iodide dosage and ICU stay time between the two groups were also compared. Results: The gastrointestinal function recovery time of patients in the treatment group was less than the control group, the difference was statistically significant (P<0.05) . From the third day of treatment, the serum cholinesterase activity of the treatment group was higher than the control group, the difference was statistically significant (P<0.05) . On the 5th day and 10th day of the treatment, the prealbumin, albumin and total protein of the treatment group were significantly higher than these indexes of the control group in the same period, the difference were statistically significant (P<0.05) . The total atropine dosage, the total pralidoxime iodide dosage and ICU stay time in the treatment group were lower than the control group, the difference were statistically significant (P<0.05) . Conclusion: Alanyl glutamine injection has a great therapeutic effect for gastrointestinal function obstacle patients caused by severe phorate poisoning.


Assuntos
Atropina/administração & dosagem , Glutamina/administração & dosagem , Inseticidas/toxicidade , Obstrução Intestinal/tratamento farmacológico , Intoxicação por Organofosfatos/tratamento farmacológico , Forato/toxicidade , Glutamina/uso terapêutico , Humanos , Índice de Gravidade de Doença , Resultado do Tratamento
9.
Zhonghua Jie He He Hu Xi Za Zhi ; 39(12): 916-923, 2016 Dec 12.
Artigo em Chinês | MEDLINE | ID: mdl-27938540

RESUMO

Objective: To study the protective effect of hydrogen inhalation on the lungs of sanitation workers exposed to haze. Methods: In this randomized, double-blind, placebo controlled clinical trial, 96 sanitation workers living in Shijiazhuang urban area were recruited during January to February, 2016. All enrolled participants were randomized to 2 groups; the treatment group inhaled H2∶O2 mixture (66.67%∶33.33%) 1 hour per day for 30 days, while the control group inhaled N2∶O2 mixture (66.67%∶33.33%) 1 hour per day for 30 days. Respiratory symptoms were evaluated and fractional exhaled nitric oxide(FeNO), biochemical indexes, lung function were measured at baseline(the 0th day) and during treatment (the 8th day, 15th day, and 30th day). Results: (1)The FeNO levels of the treatment group (16±5)×109 were lower than those of the control group(21±14)×109 on 8th day of treatment, with significant difference(F=6.94, P<0.05). (2)The levels of FEV1 were significantly higher in participants from the treatment group as compared to the control group on both 8th [(96±13)% vs(94±14)%(F=3.96, P<0.05)] and 30th day [(97±14)% vs (95±12)%(F=8.5, P<0.05)] of treatment, while PEF was also increased on 15th day [(73±15)% vs(67±18)%(F=8.68, P<0.05)]. (3)The sputum levels of MMP-12 and SOD3 were consistently lower in the treatment group as compared to the control group at each time point, and the levels of IL-10 were higher in the treatment group as compared to the control group on the 15th and 30th day. MDA and IL-2 levels were lower in the treatment group than in the control group on the 30th day(P<0.05). The sputum levels of CRP and TGF-ß1 at each time point were not different between the 2 groups (P>0.05). (4)The serum levels of IL-2 and SOD3 were lower in the treatment group as compared to the control group while IL-10 was higher than in the control group at each time point, and MMP-12 was lower in the treatment group than that in the control group on the 30th day(P<0.05). The relative ratios of CRP, TGF-ß1 and MDA in serum at each time point between the 2 groups were not significantly different (P>0.05). (5)Hydrogen inhalation improved respiratory symptoms such as cough. Conclusions: Inhalation of hydrogen gas could alleviate airway inflammation and oxidative stress of sanitation workers exposed to air pollution. There was even a significant inhibitory effect on the level of systemic inflammatory response. Importantly, inhalation of hydrogen could improve respiratory symptoms such as cough.


Assuntos
Hidrogênio/administração & dosagem , Pulmão/efeitos dos fármacos , Óxido Nítrico/administração & dosagem , Exposição Ocupacional , Estresse Oxidativo/efeitos dos fármacos , Administração por Inalação , Poluição do Ar , Asma , Testes Respiratórios/métodos , Estudos de Casos e Controles , Tosse , Método Duplo-Cego , Expiração , Feminino , Humanos , Hidrogênio/farmacologia , Interleucina-10 , Pulmão/metabolismo , Masculino , Óxido Nítrico/farmacologia , Saneamento , Escarro , Fator de Crescimento Transformador beta1
10.
Zhonghua Yi Xue Za Zhi ; 96(38): 3047-3052, 2016 Oct 18.
Artigo em Chinês | MEDLINE | ID: mdl-27784443

RESUMO

Objective: To detect the expression of heparanase (HPSE) in oral squamous cell carcinoma (OSCC) with different metastatic potentials and investigate its clinical significance. Methods: Transcriptional and translational status of HPSE in OSCC cell lines with different metastatic capacities, primary OSCC samples and their paired metastatic cancer tissues were determined using real-time polymerase chain reaction (PCR) and Western blotting analysis. Immunohistochemistry was used to evaluate HPSE expression in 131 OSCC samples. The correlation between HPSE expression pattern and clinicopathological parameters and clinical outcome in patients with OSCC were analyzed. HPSE level was reduced using HPSE-siRNAs in OSCC cell lines and its impact on cell migration and invasion was measured by scratch assay and matrigel invasion assay. Results: The mRNA and protein levels of HPSE were remarkably up-regulated in OSCC cell lines with highly metastatic capacity (P<0.000 1) and metastatic OSCC tissues (P<0.000 1). The protein levels of HPSE were strongly associated with lymph node metastasis (P<0.000 1) and tumor node metastasis stage (P=0.012). Survival analyses revealed that high HPSE expression was associated with worse overall survival (P=0.000 3). Multivariate Cox proportional analyses indicated that HPSE expression was strongly associated with clinical outcome in patients with OSCC (HR=2.203, 95% CI: 1.203-3.988, P=0.009). The siRNA-mediated silencing of HPSE could suppress the migration and invasion (P=0.008) of HN12 cells in vitro. Conclusions: The up-regulation of HPSE contributes to invasion and metastasis of OSCC. HPSE may serve as a useful biomarker for patients with OSCC.


Assuntos
Carcinoma de Células Escamosas , Movimento Celular , Glucuronidase , Humanos , Imuno-Histoquímica , Metástase Linfática , Metástase Neoplásica , RNA Mensageiro , RNA Interferente Pequeno , Reação em Cadeia da Polimerase em Tempo Real , Análise de Sobrevida , Regulação para Cima
11.
Theriogenology ; 78(4): 858-66, 2012 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-22704394

RESUMO

Domesticated animals cloned by somatic cell nuclear transfer (SCNT) generally have poor developmental competency, with many developmental abnormalities attributed to incomplete reprogramming of the nuclear genome and abnormal expression of genes important for regulation of growth and development. To investigate the molecular mechanism leading to the abnormalities of cloned animals, pathologic and histologic analyses were conducted on seven cloned cattle that were oversized at birth and had cardiac and pulmonary abnormalities. Quantitative reverse transcription (RT)-polymerase chain reaction (PCR) analysis of four imprinted genes IGF2, IGF2R, H19, and GRB10, as well as genes from related regulatory networks, were performed in liver, lung, kidney, and muscle to investigate disruption of expression. Expression of IGFBP2 was not detected in morphologically normal cloned cattle, but was detected in the liver, lung, kidney, and thymus of abnormal calves. Expression levels of IGF1 and imprinted genes IGF2 and H19 were substantially higher in these organs of abnormal cattle. In contrast, expression levels of GRB10, CTSD, and TRPV2 were substantially lower in abnormal cattle. Transcript abundance of IGFBP6 was higher in kidney, but lower in liver and lung. In conclusion, we inferred that altered expression of imprinted and related genes may be closely related to increased birth weight and pathologic changes in abnormal cloned cattle.


Assuntos
Bovinos , Clonagem de Organismos , Regulação da Expressão Gênica , Redes Reguladoras de Genes/genética , Impressão Genômica/fisiologia , Estruturas Animais/citologia , Estruturas Animais/metabolismo , Estruturas Animais/ultraestrutura , Animais , Animais Domésticos , Bovinos/embriologia , Bovinos/genética , Bovinos/fisiologia , Células Cultivadas , Clonagem de Organismos/efeitos adversos , Técnicas de Cultura Embrionária , Feminino , Impressão Genômica/genética , Proteína 2 de Ligação a Fator de Crescimento Semelhante à Insulina/genética , Proteína 2 de Ligação a Fator de Crescimento Semelhante à Insulina/metabolismo , Fator de Crescimento Insulin-Like I/genética , Fator de Crescimento Insulin-Like I/metabolismo , Técnicas de Transferência Nuclear/efeitos adversos , Gravidez
12.
Insect Mol Biol ; 18(3): 405-17, 2009 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-19523072

RESUMO

A full-length cDNA encoding a general odorant binding protein 2 (GOBP2) was cloned from the antennae of the rice striped stem borer, Chilo suppressalis (Lepidoptera: Pyralidae), by the combination of reverse transcription PCR (RT-PCR) and rapid amplification of cDNA ends PCR (RACE-PCR). The cDNA contains a 489 bp open reading frame, which encodes a 162 amino acid protein, termed as Ch. suppressalis GOBP2 (CsupGOBP2). CsupGOBP2 is similar in the number of amino acids and protein sequence to GOBP2s in other species of Lepidoptera. RT-PCR results showed that CsupGOBP2 mRNA was highly expressed in the adult antennae of both females and males, as was CsupGOBP2 protein as revealed by Western blot analysis. CsupGOBP2 expressed in Escherichia coli was purified by affinity chromatography, refolding and gel filtration from the inclusion body. Fluorescence emission spectra and competitive binding assays by using N-phenyl-1-naphthylamine as first binding ligand and odorants as potential competitors revealed that the CsupGOBP2 protein has significant affinity to cis-11-hexadecenal (Z11-16:Ald), the main component of Ch. suppressalis pheromone and to laurinaldehyd and benzaldehyde, two general plant volatile aldehydes.


Assuntos
Proteínas de Insetos/metabolismo , Mariposas/metabolismo , Receptores Odorantes/metabolismo , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , Escherichia coli , Feminino , Expressão Gênica , Proteínas de Insetos/genética , Proteínas de Insetos/isolamento & purificação , Ligantes , Masculino , Dados de Sequência Molecular , Mariposas/química , Mariposas/genética , Receptores Odorantes/genética , Receptores Odorantes/isolamento & purificação , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos
13.
J Viral Hepat ; 7(2): 104-14, 2000 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-10760040

RESUMO

Previously, we have determined that human annexin V (hAV), a Ca2+-dependent phospholipid-binding protein, and not rat AV, binds specifically to small hepatitis B surface antigen (SHBsAg), and that transfection of a rat hepatoma cell line with a construct containing the hAV gene led to hAV expression and conferred susceptibility to hepatitis B virus (HBV) infection. In this work, we have examined the effect of administration of hAV on in vitro binding of SHBsAg to human and to rat hepatocytes and on in vitro HBV infection. The results showed that hAV could bind to human as well as to rat hepatocytes. Because of this property, excess hAV was unable to prevent HBV infection in primary cultures of human hepatocytes. On the other hand, it enabled rat hepatocytes to specifically bind SHBsAg and conferred susceptibility to HBV infection. After infection of primary cultures of rat hepatocytes in the presence of hAV, HBV mRNA, covalently closed circular (ccc) DNA, replicative intermediates, hepatitis B surface antigen (HBsAg), hepatitis B core antigen (HBcAg) and secreted HBV DNA were detected. After infection in the absence of hAV, no markers of HBV replication were detected. Hence, from the present study we conclude that hAV is involved in facilitating HBV entry, leading to successful HBV infection in primary cultures of rat hepatocytes, while it is not effective in preventing HBV infection in primary cultures of human hepatocytes.


Assuntos
Anexina A5/farmacologia , Antígenos de Superfície da Hepatite B/metabolismo , Vírus da Hepatite B/fisiologia , Fígado/metabolismo , Animais , Células Cultivadas , Criopreservação , Suscetibilidade a Doenças , Hepatite B/metabolismo , Hepatite B/prevenção & controle , Vírus da Hepatite B/efeitos dos fármacos , Humanos , Fígado/citologia , Fígado/efeitos dos fármacos , Fígado/virologia , Ligação Proteica/efeitos dos fármacos , Ratos , Ratos Wistar
15.
J Viral Hepat ; 6(3): 229-36, 1999 May.
Artigo em Inglês | MEDLINE | ID: mdl-10607235

RESUMO

Mycophenolate mofetil (MMF), the morpholinoethyl ester of mycophenolic acid (MPA), is currently used as an immunosuppressive agent in kidney transplant recipients. After oral administration, MMF is hydrolysed to MPA, the active compound, which is a potent inhibitor of inosine monophosphate dehydrogenase (IMP-DH). Inhibition of this enzyme results in a depletion of the intracellular GTP and dGTP pools. MPA has been shown to inhibit the replication of a number of viruses, including arena viruses (Junin and Tacaribe), yellow fever virus, reovirus-1, parainfluenza-3 virus, Coxsackie B4 virus, Epstein-Barr virus and human immunodeficiency virus. To examine whether MPA also has an inhibitory effect on HBV replication, experiments were performed using cultures of primary human hepatocytes and HBV-transfected, HepG2 2.2.15 cells. After in vitro infection with HBV in human hepatocytes, HBV covalently-closed-circular (ccc) DNA and HBV mRNAs were detectable in the cells during the 10 days following infection. HBV DNA and hepatitis B surface antigen (HBsAg) were also secreted into the culture medium. In the presence of 10 microg ml-1 MPA (the therapeutic serum level of MPA as an immunosuppressive agent) in culture medium, HBV ccc DNA and HBV mRNAs became undetectable 5 days after treatment was started. The secretion of HBV DNA and HBsAg into the medium was also markedly reduced. No cytotoxic effect of the drug was noted during the experiments. The effect of MPA on HBV replication was abolished by the presence of guanosine (50 microg ml-1). In HepG2 2.2.15 cells (which contain an integrated tandem dimer of the HBV genome), MPA treatment had no significant inhibitory effect on the secretion of HBV DNA and HBsAg into the culture medium. HBV ccc DNA and HBV mRNAs in HepG2 2.2.15 cells were also not affected. The observed effect of MPA on HBV replication in primary human hepatocyte cultures may involve only episomal replication and may have clinical implications, especially before integration of HBV DNA into the host genome.


Assuntos
Antivirais/farmacologia , Vírus da Hepatite B/efeitos dos fármacos , Fígado/virologia , Ácido Micofenólico/farmacologia , Células Cultivadas , DNA Viral/metabolismo , Inibidores Enzimáticos/farmacologia , Guanosina/farmacologia , Antígenos de Superfície da Hepatite B/metabolismo , Vírus da Hepatite B/fisiologia , Humanos , Imunossupressores/farmacologia , Fígado/citologia , RNA Mensageiro/metabolismo , RNA Viral/metabolismo , Células Tumorais Cultivadas , Replicação Viral/efeitos dos fármacos
16.
Hepatology ; 29(2): 576-84, 1999 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-9918938

RESUMO

Previously, we have found that human liver annexin V (hA-V; in earlier reports referred as Endonexin II) is a specific hepatitis B surface antigen (HBsAg) binding protein. In this study, we demonstrate that transfection of rat hepatoma FTO 2B cells, a cell line that is not infectable by hepatitis B virus (HBV) and does not express hA-V, with a construct containing the hA-V gene, resulted in hA-V expressing cells susceptible to HBV infection. After in vitro infection, transfected FTO cells (assigned as FTO 9.1 cells) expressing hA-V in cultures were shown to contain HBV-precore/core, X mRNAs, and covalently closed circular (ccc) DNA as detected by polymerase chain reaction (PCR). The presence of HBV ccc and replicative intermediate DNA was also demonstrated by Southern blot hybridization assay. HBV DNA secreted in the culture medium was also evident as determined by quantitative branched DNA (bDNA) assay. HBsAg and hepatitis B core antigen (HBcAg) could also be detected by an immunocytochemical method in 10% to 15% of the cells at day 3 and day 5 after infection. Infectivity of in vitro-propagated HBV was demonstrated by infection of the naive FTO 9.1 cells with the culture supernatant from HBV-carrier cultures. In contrast to primary cultures of human hepatocytes and FTO 9.1 cells, primary rat and mouse hepatocytes, as well as rat hepatoma cell lines that do not express hA-V, are not susceptible to HBV infection. These findings suggest that hA-V plays a key role in the initial step of HBV infection and that the species-specific susceptibility to HBV infection and replication in hepatocytes is associated with the expression of hA-V.


Assuntos
Anexina A5/genética , Suscetibilidade a Doenças , Hepatite B , Neoplasias Hepáticas Experimentais/metabolismo , Fígado/química , Transfecção , Animais , DNA Circular/análise , DNA Viral/análise , Hepatite B/metabolismo , Hepatite B/virologia , Antígenos do Núcleo do Vírus da Hepatite B/análise , Antígenos do Núcleo do Vírus da Hepatite B/genética , Antígenos de Superfície da Hepatite B/análise , Vírus da Hepatite B/genética , Vírus da Hepatite B/fisiologia , Humanos , RNA Mensageiro/análise , RNA Viral/análise , Ratos , Ratos Wistar , Transativadores/genética , Células Tumorais Cultivadas , Proteínas Virais Reguladoras e Acessórias , Replicação Viral
17.
J Viral Hepat ; 5(6): 377-87, 1998 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-9857347

RESUMO

Studies on the pathobiology of chronic (long-term) hepatitis B virus (HBV) infection and in vitro drug testing have been hampered by the lack of appropriate systems for culturing susceptible cells chronically infected with HBV. Most of the in vitro studies of HBV replication have been performed with HBV genome-transduced cell lines. In this system, viral production is mainly the result of chromosomal replication. In an in vitro infection system, owing to medium refreshment (which leads to the removal of infectious particles necessary for the perpetuation of infection) and to trypsinization for cell passages, it is difficult, if not impossible, to maintain chronic HBV infection, despite the use of susceptible cells. To circumvent these unfavourable factors for chronic HBV infection in vitro, we cultured microcarrier-attached immortalized human hepatocytes, infected with HBV, in molecularporous (MW 12,000-14,000) membrane (dialysis) bags for a duration of 2 months. HBV covalently-closed-circular (ccc) DNA, HBV precore/core and X mRNAs were detected in the cells cultured in this system following infection until the end of the experiment (day 58), while in classical culture conditions (monolayer), markers of HBV replication were also detected. Production of hepatitis B surface antigen (HBsAg) and HBV DNA was detected and their levels in culture medium (collected at the end of experiments from the molecularporous membrane bags) were increased 2.86- and 3.28-fold respectively. Using Southern blot analysis, HBV replicative intermediates could also be demonstrated throughout the experiments. However, integrated HBV DNA was not present. In contrast, HBV ccc DNA, HBV precore/core and X mRNAs, and replicative intermediates were not demonstrable in FTO 2B rat hepatoma cells infected in the same manner in parallel experiments. This in vitro infection system, using susceptible, immortalized human hepatocytes, therefore provides a new tool for studying the long-term effect of HBV infection, mainly involving episomal replication in hepatocytes, and for drug testing.


Assuntos
Vírus da Hepatite B/fisiologia , Hepatite B Crônica/fisiopatologia , Fígado/citologia , Replicação Viral , Southern Blotting , Cápsulas , Células Cultivadas , Células Imobilizadas , Meios de Cultura , DNA Circular/análise , DNA Viral/análise , Antígenos do Núcleo do Vírus da Hepatite B/biossíntese , Antígenos de Superfície da Hepatite B/biossíntese , Humanos , Imuno-Histoquímica , Fígado/metabolismo , RNA Mensageiro/análise , RNA Viral/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Virologia/métodos
18.
J Viral Hepat ; 4(3): 145-53, 1997 May.
Artigo em Inglês | MEDLINE | ID: mdl-9181523

RESUMO

Hepatitis B virus (HBV) infection is still a major public health problem worldwide. Although much information about the molecular biology of HBV has been gained in the last decades, little is known about the mechanism of attachment and penetration of the HBV particle into human hepatocytes. The HBV envelope proteins are important for the interaction between the HBV particle and the hepatocyte plasma membrane. Although initially it was suggested that the preS2 domain could act, via polymerized human serum albumin, as an attachment site to human hepatocytes, in recent years other observations showed that the preS1 domain is probably the most important attachment site to human hepatocytes. However, controversial findings on cellular proteins for binding to the preS1 domain has been described, namely the IgA-, the IL6-, the asialoglycoprotein receptor and GAPD. Although the preS1 attachment site may be important, apo H has been shown to bind specifically to small HBsAg. Recently, we have identified human liver Annexin V as a specific small HBsAg-binding protein. In a preliminary report, the direct involvement of human Annexin V in the initial step of HBV infection has been demonstrated. A rat hepatoma cell line, which does not express human Annexin V and which is not infectable by HBV, gained the ability to become infected by HBV after transfection with human Annexin V. This result may facilitate the progress of HBV receptor research and elucidate the molecular mechanism of the initial step of HBV infection.


Assuntos
Vírus da Hepatite B/patogenicidade , Hepatite B/virologia , Fígado/virologia , Animais , Genoma Viral , Vírus da Hepatite B/genética , Vírus da Hepatite B/ultraestrutura , Humanos , Fígado/citologia , Especificidade de Órgãos , Ratos , Especificidade da Espécie , Transcrição Gênica
19.
Zhonghua Wai Ke Za Zhi ; 32(11): 662-3, 1994 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-7774403

RESUMO

26 patients with gastric stump cancer were treated by surgery plus chemotherapy or radiotherapy in our hospital from 1970 to 1992. 20 patients were qubiected to surgery plus chemotherapy 12 patients were operable six patients had simple radiotherapy. These patients were followed up to the end of 1992. Using Kaplan Meier method and log-rank test (P < 0.05), we analysed the factors effecting the survival rate and drew a conclusion that surgical treatment plus adjuvant chemotherapy is the treatment of choice, and if no-resection, we would treat it with chemotherapy rather than radiotherapy.


Assuntos
Coto Gástrico , Recidiva Local de Neoplasia/cirurgia , Neoplasias Gástricas/cirurgia , Quimioterapia Adjuvante , Seguimentos , Gastrectomia , Humanos , Neoplasia Residual , Reoperação , Neoplasias Gástricas/tratamento farmacológico
20.
Zhonghua Bing Li Xue Za Zhi ; 22(2): 72-3, 1993 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-7691424

RESUMO

Acid dyestuff, Ponceacis and Gomori method with modified silver concentration were used in demonstrating both the reticulin and collagen fibers in a satisfactory result first, and the section was then counterstained with alkalized Victoria blue, which might give a good differentiation among these different elements. It is indicated that these three staining methods might combine together and differentiate the collagen as well as reticulin fibers from fibrin in the same section simultaneously. The staining was known to be rather stable.


Assuntos
Colágeno/análise , Tecido Conjuntivo/química , Fibrina/análise , Reticulina/análise , Animais , Humanos , Coloração e Rotulagem
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