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1.
Zhonghua Kou Qiang Yi Xue Za Zhi ; 59(8): 824-833, 2024 Jul 22.
Artigo em Chinês | MEDLINE | ID: mdl-39036915

RESUMO

Objective: To investigate the expression of histone demethylase, Jumonji domain-containing protein 3 (JMJD3), in inflammatory periodontal tissues and its potential mechanism for the regulation of periodontitis. Methods: The results of single-cell sequencing of periodontal tissues published in the Gene Expression Omnibus (GEO) database in 2022 were analyzed. Nine gingival samples each from healthy and inflamed periodontal patients were collected during periodontal surgery or tooth extractions for immunohistochemical staining and real-time fluorescence quantitative PCR (RT-qPCR). Mice periodontitis models were constructed, and the experimental groups were: healthy control+saline group, silk ligation+saline group, silk ligation+GSK-J4(inhibitor of JMJD3) group. Lipopolysaccharide (LPS) derived from Porphyromonas gingivalis (Pg) (Pg-LPS) was used to mimic the periodontal inflammatory microenvironment. The macrophages were treated with small interfering RNA (siRNA) targeting Jmjd3 and the JMJD3 inhibitor GSK-J4. siRNA transfection experiments were grouped into the following: the NC group (negative control sequence transfection group), the siRNA-Jmjd3 group, the NC+LPS group, siRNA-Jmjd3+LPS group. Inhibitor experiments were grouped as dimethyl sulfoxide (DMSO) group, GSK-J4 group, DMSO+LPS group, GSK-J4+LPS group. Western blotting and immunofluorescence staining were used to explore the effects of JMJD3 on macrophage polarization and periodontal inflammation in the in vivo and in vitro settings. Results: RT-qPCR results showed that JMJD3 expression in gingival tissues of periodontitis patients (1.97±0.91) was significantly higher than that in healthy gingival tissues (1.00±0.33) (t=2.45, P=0.048). RT-qPCR results of in vitro experiments showed that either siRNA knockdown of JMJD3 or inhibition of JMJD3 using GSK-J4 promoted M1 polarization and inhibited M2 polarization in macrophages under inflammatory environment: the expression of arginase I (Arg 1) in the NC+LPS group (0.90±0.06) was significantly higher than that in the siRNA-Jmjd3+LPS group (0.61±0.11) (P<0.01); the expression of interleukin (Il)-6, Il-1ß, and tumor necrosis factor alpha (Tnf-α) in the NC+LPS group (8.50±0.16, 5.56±0.20, 3.44±0.16) were significantly lower than those in the siRNA-Jmjd3+LPS group (14.63±0.48, 8.55±0.10, 11.72±0.16) (P<0.01). The expression of Arg-1, Ym1, Il-10 in the DMSO+LPS group (0.82±0.01, 0.35±0.16, 1.47±0.11) were significantly higher (P<0.01) than the GSK-J4+LPS group (0.55±0.03, 0.22±0.21, 0.51±0.11); the expression of Il-6, Il-1ß, and Tnf-α in the DMSO+LPS group (2.03±0.13, 3.63±0.14, 4.06±0.03) were significantly lower than the GSK-J4+LPS group (2.69±0.16, 15.04±1.15, 4.36±0.10) (P<0.01). The results of the in vivo experiments revealed that inhibition of JMJD3 exacerbated bone loss in experimental periodontitis mice, increased macrophage M1 polarization, and decreased M2 polarization in inflamed periodontal tissues. The buccal cemento-enamel junction (CEJ)-alveolar bone crest (ABC), palatal CEJ-ABC, as well as the ratio of M1/M2 type macrophages were significantly lower in the silk ligation+saline group [(0.26±0.03), (0.24±0.01) mm, 0.35±0.10) than in the silk ligation+GSK-J4 group [(0.34±0.04), (0.30±0.05) mm, 2.50±0.58) (t=3.65, P=0.006; t=2.67, P=0.049; t=7.31, P=0.004; respectively). Conclusions: Single-cell sequencing as well as the in vitro and in vivo experiments verified that JMJD3 expression was upregulated in periodontitis periodontal tissues. JMJD3 may exert a protective role in periodontitis by regulating macrophage polarization, thereby inhibiting alveolar bone destruction associated with the periodontitis.

2.
Zhonghua Liu Xing Bing Xue Za Zhi ; 44(10): 1567-1574, 2023 Oct 10.
Artigo em Chinês | MEDLINE | ID: mdl-37875443

RESUMO

Objective: To evaluate the level of psychological distress among Chinese children and adolescents and analyze its lifestyle influencing factors. Methods: Data were obtained from the 2019 Chinese National Survey on Students' Constitution and Health. A lotal of 120 285 Han Chinese children and adolescents aged 9-18 years with complete information on the psychological distress scale and lifestyle factors were selected, including 58 432 boys and 61 853 girls. The Kessler Psychological Distress Scale (K10) measured psychological distress, and lifestyles such as physical activity, sedentary behavior, diet, and sleep were also investigated. K10 scores of different genders were compared using the t-test, and the levels of psychological distress were compared using the χ2 test. Logistic regression was used to analyze lifestyle risk factors associated with high psychological distress, and multiple linear regression was used to find the relationship between K10 scores and lifestyle scores. Results: The average K10 score for Han Chinese children and adolescents aged 9-18 years was 21.25±7.35, with girls (21.43±7.35) scoring higher than boys (21.06±7.36), the difference was statistically significant (t=8.72, P<0.001). The rate of high psychological distress was 29.81%, with girls (31.08%) reporting higher rates than boys (28.46%), the difference was statistically significant (χ2=98.54,P<0.001). 56.10% of children and adolescents have unhealthy lifestyles, with girls (58.77%) reporting higher rates than boys (53.27%), the difference was statistically significant (χ2=368.53,P<0.001). Except for insufficient outdoor activities for girls (P=0.128), lifestyles such as insufficient physical activity, insufficient muscle-and-bone exercises, long screen time, not eating breakfast, eggs and dairy products every day, drinking sugary beverages once or more per day, and not having enough sleep are all risk factors for high psychological distress (all P<0.001). For every additional healthy lifestyle score, the K10 score decreased by 0.98 [ß=-0.98 (95%CI: -1.01- -0.95)] points (P<0.001). K10 scores in each region negatively correlate with lifestyle scores (all P<0.001). Among them, the K10 score in the eastern region showed the slightest decrease as the lifestyle score increased, while the western region showed the most decrease. Conclusions: The prevalence of psychological distress and unhealthy lifestyle in Chinese children and adolescents are high and interrelated. Compared those with healthy lifestyles, children and adolescents with unhealthy lifestyles are at greater risk of high psychological distress. Therefore, promoting healthy lifestyles for children and adolescents may be one of the important ways to improve their mental health.


Assuntos
Estilo de Vida , Sono , Humanos , Masculino , Feminino , Adolescente , Criança , Exercício Físico , Saúde Mental , Estilo de Vida Saudável
4.
Lin Chuang Er Bi Yan Hou Tou Jing Wai Ke Za Zhi ; 31(24): 1892-1895, 2017 Dec 20.
Artigo em Chinês | MEDLINE | ID: mdl-29798311

RESUMO

Objective:To discuss similarities and differences in clinical features and laboratory indexes between patients with flat descending type sudden hearing loss and those with total hearing loss. Method:The clinical data of 123 patients with full frequencies hearing loss were retrospectively analyzed. The differences in clinical features and laboratory tests(platelet, coagulation series, D-dimer, blood lipids, hemorheology) between patients with flat descending hearing loss and those with total hearing loss were analyzed by gender, age and ear side, treatment time, concomitant symptom (tinnitus, dizziness), original underlying diseases (hypertension, diabetes), etc. Result:In the clinical features,among 51 flat descending cases, the ratio of male and female was 2.401:1; among 72 total hearing loss cases, the ratio of men and women ratio was 1.058:1 (P<0.05). Among two groups of patients,the majority received treatment within 7 days, among whom 66.7% were flat descending population, and 83.3% were total hearing loss population (P<0.05). Flat descending population with dizziness only accounted for 35.3% while this figure was up to 70.8% when it came to total hearing loss patients (P<0.01). Two groups showed no differences in age, ear side, tinnitus, the original underlying diseases (hypertension, diabetes). In the laboratory tests, the total hearing loss population overtopped the plat descending population in PLT and PCT (P<0.05), while falling below the plat descending population in APTT (P<0.01). Two groups showed no differences in other indicators of platelet and coagulation series and laboratory data of D-dimer, blood lipids, hemorheology. Conclusion:Compared with flat descending sudden hearing loss, sudden total hearing loss more frequently happened to females who also were accompanied by dizziness. The treatment rate within 7 days was high and the patients with hypercoagulable state accounted for a higher proportion.


Assuntos
Surdez/diagnóstico , Perda Auditiva Neurossensorial/diagnóstico , Perda Auditiva Súbita/diagnóstico , Surdez/complicações , Feminino , Perda Auditiva Neurossensorial/complicações , Perda Auditiva Súbita/complicações , Testes Auditivos , Humanos , Masculino , Zumbido/etiologia , Vertigem/etiologia
5.
Poult Sci ; 94(11): 2686-94, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26500268

RESUMO

The objective of the present study was to investigate the effects of dietary supplemental Zinc (Zn) source and level on antioxidant ability and fat metabolism-related enzymes of broilers. Dietary treatments included the Zn-unsupplemented corn-soybean meal basal diet (control) and basal diets supplemented with 60, 120, or 180 mg Zn/kg as Zn sulfate, Zn amino acid chelate with a weak chelation strength of 6.5 quotient of formation (Qf) (11.93% Zn) (Zn-AA W), Zn proteinate with a moderate chelation strength of 30.7 Qf (13.27% Zn) (Zn-Pro M), or Zn proteinate with an extremely strong chelation strength of 944.0 Qf (18.61% Zn) (Zn-Pro S). The results showed that dietary supplemental Zn increased (P < 0.01) Zn contents in the liver, breast, and thigh muscles of broilers, and up-regulated mRNA expressions of copper and Zn containing superoxide dismutase (CuZnSOD) and metallothioneins (MT) in the liver (P < 0.01) and thigh muscle (P < 0.05), and also enhanced (P < 0.05) CuZnSOD activities in the breast and thigh muscles, which exerted antioxidant ability and a decreased malondialdehyde (MDA) level in the liver (P < 0.01) and breast and thigh muscles (P < 0.05) of broilers. Furthermore, supplemental Zn increased activities of malate dehydrogenase (MDH) and lipoprotein lipase (LPL) in the abdominal fat (P < 0.05), and fatty acid synthetase (FAS) and LPL in the liver (P < 0.01), which were accompanied with up-regulation (P < 0.01) of the mRNA expressions levels of these enzymes in the abdominal fat and liver of broilers. Dietary Zn source, and an interaction between Zn source and level, had no effects on any measurements. It is concluded that dietary Zn supplementation improved Zn status and resulted in promoting antioxidant ability and activities and gene expressions of fat metabolism-related enzymes of broilers regardless of Zn source and level, and the addition of 60 mg Zn/kg to the corn-soybean meal basal diet (a total dietary Zn of approximately 90 mg/kg) was appropriate for improving the above aspects of broilers.


Assuntos
Antioxidantes/metabolismo , Galinhas/metabolismo , Suplementos Nutricionais , Enzimas/metabolismo , Gorduras/metabolismo , Zinco/metabolismo , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Animais , Dieta/veterinária , Relação Dose-Resposta a Droga , Masculino , Distribuição Aleatória , Reação em Cadeia da Polimerase em Tempo Real/veterinária
6.
Braz. j. med. biol. res ; 48(7): 654-664, 07/2015. graf
Artigo em Inglês | LILACS | ID: lil-751344

RESUMO

Recent evidence indicates that a deficiency of 1,25-dihydroxyvitamin D3 (1,25[OH]2D3) may influence asthma pathogenesis; however, its roles in regulating specific molecular transcription mechanisms remain unclear. We aimed to investigate the effect of 1,25(OH)2D3 on the expression and enzyme activity of histone deacetylase 2 (HDAC2) and its synergistic effects with dexamethasone (Dx) in the inhibition of inflammatory cytokine secretion in a rat asthma model. Healthy Wistar rats were randomly divided into 6 groups: control, asthma, 1,25(OH)2D3 pretreatment, 1,25(OH)2D3 treatment, Dx treatment, and Dx and 1,25(OH)2D3 treatment. Pulmonary inflammation was induced by ovalbumin (OVA) sensitization and challenge (OVA/OVA). Inflammatory cells and cytokines in the bronchoalveolar lavage (BAL) fluid and histological changes in lung tissue were examined. Nuclear factor kappa B (NF-κB) p65 and HDAC2 expression levels were assessed with Western blot analyses and quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR). Enzyme activity measurements and immunohistochemical detection of HDAC2 were also performed. Our data demonstrated that 1,25(OH)2D3 reduced the airway inflammatory response and the level of inflammatory cytokines in BAL. Although NF-κB p65 expression was attenuated in the pretreatment and treatment groups, the expression and enzyme activity of HDAC2 were increased. In addition, 1,25(OH)2D3 and Dx had synergistic effects on the suppression of total cell infusion, cytokine release, and NF-κB p65 expression, and they also increased HDAC2 expression and activity in OVA/OVA rats. Collectively, our results indicated that 1,25(OH)2D3 might be useful as a novel HDAC2 activator in the treatment of asthma.


Assuntos
Animais , Masculino , Asma/tratamento farmacológico , Calcitriol/farmacologia , /efeitos dos fármacos , NF-kappa B/efeitos dos fármacos , Vitaminas/farmacologia , Asma/induzido quimicamente , Western Blotting , Líquido da Lavagem Broncoalveolar/química , Contagem de Células , Calcitriol/uso terapêutico , Citocinas/análise , Citocinas/efeitos dos fármacos , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , /metabolismo , Imuno-Histoquímica , Pulmão/química , Pulmão/efeitos dos fármacos , NF-kappa B/análise , Ovalbumina , Ratos Wistar , Reação em Cadeia da Polimerase em Tempo Real , Reprodutibilidade dos Testes , Resultado do Tratamento , Vitaminas/uso terapêutico
7.
Eur Rev Med Pharmacol Sci ; 17(3): 349-55, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23426538

RESUMO

BACKGROUND: Severe acute pancreatitis (SAP) is a serious systemic disease with a sustained high mortality rate. Extensive evidence has shown that gut barrier dysfunction plays a critical role in the pathophysiology of SAP. AIM: Investigating the role of intestinal mucosa oxidative stress in gut barrier dysfunction of SAP. MATERIALS AND METHODS: Twenty-four BALB/c mice were randomly divided into two groups with twelve mice each group. The SAP group mice received six intraperitoneal injections of cerulein (50 µg/kg) at 1-hour intervals, then given one intraperitoneal injection of 10 mg/kg lipopolysaccharide (LPS from E. coli) for inducing SAP. Normal saline was given to the mice of control group. The animals of each group were averaged to two batches. Four and eight hours after the final injection, respectively, mice were anesthetized and blood and tissue samples were harvested for examination. The pathological changes of pancreas and gut were observed and scored. The serum levels of diamine oxidase (DAO), amylase and tumor necrosis factor-alpha (TNF-α) were measured. The contents of malondialdehyde (MDA) and reduced glutathione (GSH) and activity of superoxide dismutase (SOD) and xanthine oxidase (XO) in gut mucosa were detected. In gut mucosa, the caspase-3 activity was measured and the cell apoptosis and apoptosis index (AI) were determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. The data were analyzed by ANOVA and t-test. RESULTS: At four and eight hours after SAP induction, the SAP group mice had significantly higher pancreatic and gut pathological scores (p < 0.01) and increased serum levels of amylase (p < 0.05), DAO and TNF-α (p < 0.01) and increased MDA contents and XO activity of gut mucosa (p < 0.01) compared with those of control mice. There were significantly lower GSH contents (p < 0.05) and SOD activity (p < 0.01) of gut mucosa in the SAP mice. It was also observed that the gut mucosa cells of SAP mice had significantly higher caspase-3 activity and apoptosis index (p < 0.01). CONCLUSIONS: In SAP, waterfall-style release of inflammatory factors such as TNF-α led to ischemia-reperfusion injury of gut mucosa which resulted in serious oxidative stress and activation of caspase-3 pathway and severe apoptosis of gut mucosa. Therefore, intestinal mucosal oxidative stress may play an important role in the mechanism of gut barrier dysfunction.


Assuntos
Mucosa Intestinal/fisiopatologia , Estresse Oxidativo , Pancreatite/fisiopatologia , Traumatismo por Reperfusão/fisiopatologia , Doença Aguda , Análise de Variância , Animais , Apoptose , Caspase 3/metabolismo , Ceruletídeo/toxicidade , Modelos Animais de Doenças , Marcação In Situ das Extremidades Cortadas , Mediadores da Inflamação/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Distribuição Aleatória , Índice de Gravidade de Doença , Fator de Necrose Tumoral alfa/metabolismo
8.
Cancer Gene Ther ; 15(3): 173-82, 2008 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-18157145

RESUMO

Oncolytic adenoviruses, also called conditionally replicating adenoviruses (CRADs), have been widely applied in cancer gene therapy. However, the construction of CRADs is still time-consuming. In this study, we attempted to establish a simplified method of generating CRADs based on AdEasy system. A novel plasmid pTE-TPE-GM was constructed, containing sequentially positioned promoter of telomerase reverse transcriptase (TERTp), coding sequence of E1A gene, promoter of E1B gene, granulocyte-macrophage colony-stimulating factor (GM-CSF) gene, internal ribosome entry site sequence and coding sequence of E1B55K gene. The CRAD-generating system reported here include three plasmids: pTE-TPE-GM, pShuttle-CMV and AdEasy-1, one Escherichia coli strain BJ5183, and the packaging cell line 293. Using this system, an oncolytic adenovirus carrying B7-1 (CD80) and GM-CSF genes was successfully constructed and designated as Ad-CD80-TPE-GM. The expression of GM-CSF increased more than 9000 times in tumor cell lines infected by Ad-CD80-TPE-GM at a multiplicity of infection (MOI) of 5, compared with the cells infected by replication-defective control virus. Similarly, the expression of CD80 also increased 9-140 times. Ad-CD80-TPE-GM selectively replicates in TERT-positive tumor cells, and the progeny viruses can reach up to 375 infection units (IU) per cell. In vitro study showed that the Ad-CD80-TPE-GM induced an obvious oncolytic effect at MOI of 0.1, and killed about 80% TERT-positive tumor cells within 7 days at an MOI of 1. The antitumor effect of this vector was also investigated in Hep2 xenograft model of nude mice, and the tumor inhibition rate reached 74% at day 30 after the administration with a total dose of 1 x 10(9) IU Ad-CD80-TPE-GM. Intratumoral injection of Ad-CD80-TPE-GM slightly induced neutralizing antibody against the oncolytic adenovirus in nude mice, which might contribute to the virus clearance in vivo. In conclusion, we successfully constructed an oncolytic CRAD carrying GM-CSF and CD80 gene. More importantly, this system can be modified to generate novel transcriptionally regulated CRADs with different tissue-specific promoters or transgenes.


Assuntos
Adenoviridae/genética , Vetores Genéticos/genética , Vírus Oncolíticos/genética , Transgenes/genética , Ensaios Antitumorais Modelo de Xenoenxerto , Animais , Antígeno B7-1/genética , Linhagem Celular , Linhagem Celular Tumoral , Sobrevivência Celular/genética , Sobrevivência Celular/fisiologia , Terapia Genética/métodos , Fator Estimulador de Colônias de Granulócitos e Macrófagos/genética , Células HT29 , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Terapia Viral Oncolítica/métodos , Vírus Oncolíticos/fisiologia , Replicação Viral/genética , Replicação Viral/fisiologia
9.
Se Pu ; 18(6): 574-6, 2000 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-12541756

RESUMO

The experimental data were analyzed by regression analytical method and the relation between the quantity of impurities and the boiling range of benzene was demonstrated. It was proved that when the quantity of hydrocarbon impurities was within certain limits, the quality of benzene fractions can be determined by analyzing the hydrocarbon impurities in it. So the distillation method for analyzing benzene boiling range may be replaced by capillary chromatography for analyzing the hydrocarbon impurities. The practical applications support the above suggestion.

10.
Acta andin ; 4(2): 143-8, 1995. tab
Artigo em Inglês | LILACS | ID: lil-187064

RESUMO

En China, las altiplanicies de Qinghai en el Tibet son las más altas del mundo, y en ellas residen los tibetianos nativos y los inmigrantes chinos (Han). Este artículo se ocupa de las enfermedades producidas por la altura. Existen tres problemas serios producidos por la altura. Primero, el edema pulmonar de altura (HAPE); se observa una alta incidencia de HAPE en los recién llegados a la altura, y por el contrario una menor incidencia en los nativos de altura que reascienden a ella, lo que difiere con los reportes en Norte América y en los Andes. Segundo, la enfermedad cardiaca de altura (HAHD). Los niños y los infantes son los que se encuentran especialmente en riesgo. La mayoría de infantes afectados por esta enfermedad son de origen Han, y presentan insuficiencia cardiaca congestiva severa debido a hipertension pulmonar dentro de los pocos meses de nacido o de arribo a la altura. Las autopsias muestran una hipertrofia ventricular derecha y un engrosamiento de la muscular de las arterias pulmonares periféricas. La mortalidad en promedio fue de 15 por ciento, tal que la HAHD infantil es una emfermedad fatal. Tercero, la enfermedad de Monge o mal de Montaña Crónico (CMS). En los indígenas tibetianos se han observado 15 casos de CMS entre 1991-1993. Los datos epidemiológicos y clínicos muestran que la CMS existe en la altiplanicie de Qinghai-Tibet.


Assuntos
Humanos , Altitude , Doença da Altitude/etnologia , Insuficiência Cardíaca/etiologia , Doença da Altitude/epidemiologia , Doença da Altitude/etiologia , Doença da Altitude/fisiopatologia , Insuficiência Cardíaca/diagnóstico , Edema Pulmonar/epidemiologia , Edema Pulmonar/etiologia , Edema Pulmonar/patologia
11.
Carcinogenesis ; 15(12): 2859-66, 1994 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-8001247

RESUMO

The tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1- butanone (NNK) is a potential human carcinogen that is known to be metabolized to DNA-reactive intermediates by the cytochromes P450. We have examined the nature of NNK's DNA damaging effects in a mammalian cell system expressing a specific human cytochrome P450 (2A6) and containing a target gene for mutagenesis. Human CYP2A6, which is known to activate NNK to a mutagen, was lipofected via a retroviral vector into the Chinese hamster ovary AS52 cell line, which contains the bacterial gpt gene and can be mutated to 6-thioguanine resistance. AS52 cells expressed negligible CYP2A6-specific coumarin 7-hydroxylase activity (0.7 pmol/mg protein/min), while a CYP2A6 transfected clone (AS52-E8) expressed 30 pmol/mg protein/min. Both cell lines were equally sensitive to the cytotoxic and mutagenic effects of the direct-acting mutagen ethylmethanesulfonate; however, only the AS52-E8 cells exhibited a dose-dependent increase in cytotoxicity and mutant frequency upon treatment with NNK. At the highest NNK dose (1200 micrograms/ml), the mutant frequency in AS52-E8 cells was 14-fold (339 x 10(-6)) greater than the spontaneous frequency of 24 x 10(-6). Ninty-eight mutant clones were isolated following NNK treatment. Based on PCR analysis, 21 clones contained deletions/rearrangements and 77 were putative point mutants. Sequencing potential point mutants showed that 81% contained G:C to A:T transitions. Four of six G:C to A:T hotspots were at the second G of the GGT motif, which is the motif and major mutation found in codon 12 of Ki-ras from NNK-induced lung tumors in strain A mice. Since NNK may be metabolized via different pathways to pyridyloxobutylate or methylate DNA, the data suggest that methylation damage causes the major mutagenic events in AS52-E8 cells when NNK is activated by human CYP2A6.


Assuntos
Hidrocarboneto de Aril Hidroxilases , Sistema Enzimático do Citocromo P-450/metabolismo , Oxigenases de Função Mista/metabolismo , Nitrosaminas/metabolismo , Proteínas , Sequência de Aminoácidos , Animais , Proteínas de Bactérias/genética , Sequência de Bases , Biotransformação , Células CHO , Linhagem Celular , Cricetinae , Citocromo P-450 CYP2A6 , Sistema Enzimático do Citocromo P-450/genética , Proteínas de Escherichia coli , Genes Reporter/efeitos dos fármacos , Genes ras , Hipoxantina Fosforribosiltransferase/genética , Oxigenases de Função Mista/genética , Dados de Sequência Molecular , Testes de Mutagenicidade , Nitrosaminas/toxicidade , Pentosiltransferases , Reação em Cadeia da Polimerase , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Tetra-Hidrofolato Desidrogenase/genética
12.
Carcinogenesis ; 14(7): 1421-7, 1993 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-8330360

RESUMO

In order to develop more efficient in vitro systems for the study of pro-mutagenic or pro-carcinogenic chemicals, we have produced transgenic C3H/10T1/2 cell lines expressing human cytochrome P450 (CYP) 2A6. A retroviral vector containing the cDNA was packaged in psi-2 cells, and used to infect C3H/10T1/2 cells. From 100 G418-resistant clones initially isolated, three cell lines were chosen for further study based upon their morphologies, growth rates and CYP2A6-dependent coumarin 7-hydroxylase activities. Infected clone 10T1/2-04, like the 10T1/2 cells, had no detectable CYP2A6 enzyme activity, while clones 10T1/2-10 and 10T1/2-29 had microsomal CYP2A6 enzyme activities within the range found in human liver microsomes. CYP2A6 protein levels were in agreement with the observed enzyme activities. Southern blots revealed that cells from clone 10T1/2-04 contained a vector lacking the CYP2A6 cDNA, while cells from clones 10T1/2-10 and 10T1/2-29 contained multiple full-length inserts. Southern analysis also indicated the presence of an endogenous CYP2A6 ortholog in the four cell lines. All cell lines exhibited about equal sensitivity to induction of cytotoxicity and conversion to ouabain resistance by the direct acting mutagen N-methyl-N'-nitro-N-nitrosoguanidine. The four lines were also about equally sensitive to transformation by benzo[a]pyrene, a chemical requiring metabolic activation. However, only clones 10T1/2-10 and 10T1/2-29, which express CYP2A6 activity, were mutated and morphologically transformed by the tobacco specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone.


Assuntos
Hidrocarboneto de Aril Hidroxilases , Transformação Celular Neoplásica , Sistema Enzimático do Citocromo P-450/metabolismo , Oxigenases de Função Mista/metabolismo , Nitrosaminas/toxicidade , Retroviridae/genética , Animais , Biotransformação , Southern Blotting , Western Blotting , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Transformação Celular Neoplásica/genética , Clonagem Molecular , Citocromo P-450 CYP2A6 , Sistema Enzimático do Citocromo P-450/genética , Vetores Genéticos , Humanos , Camundongos , Camundongos Endogâmicos C3H , Oxigenases de Função Mista/genética , Nitrosaminas/metabolismo , Nitrosaminas/farmacocinética
13.
Zhonghua Zhong Liu Za Zhi ; 15(4): 300-2, 1993 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-8174469

RESUMO

From August 1974 to January 1987, 650 cases of moderately and advanced esophageal carcinoma were treated with a combination of chemotherapy and Rabdosia rubescens or Rabdosia rubescens and/or tradition chinese medicinal prescription. After treatment, 40 patients survived for over 5 years (5-year survival rate 6.15%): 32 for over 6 years, 23 for more than 10 years, 5 for more than 15 years and 20 die of tumors (16 cases) or other diseases (4 cases). There were 20 patients who are still living and some of them have been living for more than 18 years. Analyzing the data, it is believed that the age, the state of activity, the length of illness, the effectiveness of primary treatment, the multi-course extensive therapy, long-term maintenance treatment, etc, are all important factors affecting the results of drug treatment.


Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Carcinoma de Células Escamosas/tratamento farmacológico , Medicamentos de Ervas Chinesas/uso terapêutico , Neoplasias Esofágicas/tratamento farmacológico , Adulto , Idoso , Carcinoma de Células Escamosas/mortalidade , Neoplasias Esofágicas/mortalidade , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Taxa de Sobrevida
14.
J Formos Med Assoc ; 91(3): 247-51, 1992 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-1354683

RESUMO

The hallmark of chronic myelogenous leukemia (CML) is the Philadelphia chromosome (Ph1) which is caused by a translocation of the c-abl gene from chromosome 9 to the breakpoint cluster region (bcr) on chromosome 22. Polymerase chain reaction (PCR) can be used to detect the chimeric bcr/c-abl mRNA as evidence of the translocation. We applied a very simple and quick method of isolating cytoplasmic RNA, as well as reverse transcription and PCR in detecting bcr/c-abl mRNA of seven CML patients in various stages. Our results showed that only a small amount of either peripheral blood or bone marrow material was required for the expression of the bcr/c-abl mRNA. This is a very fast and time-saving method since a one-step method of cytoplasmic RNA isolation is used instead of the several steps in total RNA isolation as published in other literature.


Assuntos
Proteínas de Fusão bcr-abl/genética , Leucemia Mielogênica Crônica BCR-ABL Positiva/genética , Reação em Cadeia da Polimerase , RNA Mensageiro/análise , Translocação Genética , Sequência de Bases , Humanos , Dados de Sequência Molecular
15.
Am J Obstet Gynecol ; 166(1 Pt 1): 35-40, 1992 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-1310201

RESUMO

OBJECTIVE: The objective of this study was to examine the possibility of intrauterine human papillomavirus infection of fetuses by transplacental transmission of human papillomavirus before delivery. STUDY DESIGN: Specimens of cervicovaginal cells and peripheral blood mononuclear cells were obtained from 52 consecutive pregnant women in the third trimester of pregnancy. Cord blood specimens were also obtained from the neonates born to these mothers. Presence of human papillomavirus types 16 and 18 deoxyribonucleic acid was analyzed by an in vitro enzymatic deoxyribonucleic acid amplification method. RESULTS: Human papillomavirus type 16 deoxyribonucleic acid was found in 6 (11.5%) cervicovaginal and in 9 (17.3%) peripheral blood mononuclear cell specimens. Seven cord blood specimens from neonates born to mothers who were positive for peripheral blood mononuclear cell human papillomavirus type 16 deoxyribonucleic acid were found to contain human papillomavirus type 16 deoxyribonucleic acid. One cervicovaginal and two peripheral blood mononuclear cell specimens contained human papillomavirus type 18 deoxyribonucleic acid, but none of the cord blood specimens contained human papillomavirus type 18 deoxyribonucleic acid. CONCLUSION: These results seem to suggest possible transplacental transmission of the virus and the potential association of such transmission with the status of human papillomavirus in peripheral blood mononuclear cells.


Assuntos
Papillomaviridae , Complicações Infecciosas na Gravidez/microbiologia , Infecções Tumorais por Vírus/transmissão , Sequência de Bases , Colo do Útero/microbiologia , DNA Viral/análise , Feminino , Sangue Fetal/microbiologia , Humanos , Leucócitos Mononucleares/microbiologia , Dados de Sequência Molecular , Papillomaviridae/genética , Reação em Cadeia da Polimerase , Gravidez , Terceiro Trimestre da Gravidez , Infecções Tumorais por Vírus/microbiologia , Vagina/microbiologia
16.
Zhonghua Bing Li Xue Za Zhi ; 19(2): 90-3, 1990 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-2390796

RESUMO

97 cases of thyroid carcinoma originated from follicular epithelium were investigated by using histological and immunohistochemical techniques with special reference to lectin distribution. According to the WHO histological typing of thyroid tumours, these cases were divided into three categories as follows: papillary carcinoma of thyroid (PCT) 56, follicular carcinoma of thyroid (FCT) 31 and undifferentiated carcinoma of thyroid (UCT) 10. Results showed that three different kinds of thyroid carcinoma presented various hormone function and distribution of lectins. The positive rate of Tg immunoreactivity was significantly different between these three kinds of tumour, i.e. PCT greater than FCT greater than UCT. Additionally, the positive rate of T4 and T3 immunoreactivity was lower than that of Tg. Some Gastrin, SS and calcitonin positive cells were also recognized in carcinoma of thyroid. Lectin--binding rate of WGA, PNA, SBA and UEA to 97 cases of thyroid carcinoma and 9 cases of normal thyroid tissue revealed that different lectin had a selective binding activity to various types of thyroid carcinoma and normal thyroid cells. From the data obtained, it seemed that the morphological differentiation of thyroid carcinoma was in correspondence with difference of function, and the extent of cell differentiation may be closely related to the biological behavior of the tumour.


Assuntos
Carcinoma Papilar/análise , Lectinas/análise , Tireoglobulina/análise , Neoplasias da Glândula Tireoide/análise , Adenocarcinoma/análise , Carcinoma/análise , Humanos , Imuno-Histoquímica , Tiroxina/análise , Tri-Iodotironina/análise
17.
Contraception ; 37(2): 191-5, 1988 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-3370992

RESUMO

Gossypol acetic acid has been shown to have the effect of increasing the SCE frequencies in the spermatogonial cells of mice. This study further confirmed the genotoxic effects of gossypol acetic acid by investigating the dose-response relationships of gossypol acetic acid-induced SCEs in spermatogonial cells and bone marrow cells of mice. Twelve or over 12 mg/kg body weight of the dosage per day could increase the SCE frequencies of both bone marrow cells and spermatogonial cells. The increase of SCE frequency appeared to be directly correlated with the doses used at significant levels (p less than 0.001). Regression lines indicated that, in spite of cell type variation, the SCE induced was proportional to the dose of gossypol acetic acid.


Assuntos
Medula Óssea/efeitos dos fármacos , Anticoncepcionais Masculinos/farmacologia , Gossipol/análogos & derivados , Troca de Cromátide Irmã/efeitos dos fármacos , Espermicidas/farmacologia , Espermatogônias/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Animais , Relação Dose-Resposta a Droga , Gossipol/farmacologia , Masculino , Camundongos
18.
Zhonghua Zhong Liu Za Zhi ; 8(4): 297-9, 1986 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-2428570

RESUMO

One hundred and fifteen patients with inoperable esophageal carcinoma were treated by either chemotherapy alone or chemotherapy plus Rabdosia rubescens. In group A, out of 31 patients treated with pingyangmycin (P) and nitrocaphane (N), 10 (32.3%) responded to the treatment. Among them, 2 showed partial response (greater than 50% tumor regression) and 8 minimal response (greater than 50% tumor regression). In group B, out of 84 patients treated with PN plus Rabdosia rubescens, 59 (70.2%) responded. Of them, 10 showed complete response (100% tumor regression), 16 partial response and 33 minimal response. the one-year survival rates of group A and B were 13.6% and 41.3%. Statistical significance was present in these two groups both in the response rate and one-year survival rate. As regards the drug toxicity, there was no significant difference between these two groups. Alopecia, anorexia, nausea and hyperpyrexia occurred in more than 30% of patients. Mild leukopenia and thrombocytopenia and interstitial pneumonia were noted in some patients, and two patients died of toxicity in the lungs.


Assuntos
Carcinoma de Células Escamosas/tratamento farmacológico , Neoplasias Esofágicas/tratamento farmacológico , Medicina Tradicional Chinesa , Medicina Tradicional do Leste Asiático , Plantas Medicinais , Adulto , Idoso , Antibióticos Antineoplásicos/uso terapêutico , Antineoplásicos/uso terapêutico , Bleomicina/uso terapêutico , Sinergismo Farmacológico , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Compostos de Mostarda Nitrogenada/uso terapêutico , Extratos Vegetais/uso terapêutico
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