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1.
Heliyon ; 10(1): e23758, 2024 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-38226234

RESUMO

Objective: In this study, we aimed to investigate whether age first had sexual intercourse (AFSI) and lifetime number of sexual partners (LNSP) have a direct causal effect on cervical cancer by Mendelian randomization (MR) analysis. Methods: Four approaches were used for MR Analysis, including MR-Egger, weighted method, weighted median, and inverse variance weighted (IVW). MR Pleiotropy RESidual Sum and Outlier (MR-PRESSO) as well as MR-Egger regression analysis were conducted to detect whether there was pleiotropy between IVs and outcome, and the outlier SNPs can be detected by MR-PRESSO. The presence or absence of heterogeneity among IVs was suggested according to Cochran's Q statistic. Leave-one-out sensitivity analysis was performed to identify and remove SNPs which could independently change the results. We corrected the results using Bonferroni correction. Results: From the results of IVW, AFSI had a negative effect on cervical cancer (OR = 0.996, 95 % CI: 0.995, 0.998 P = 1.70E-07), which still persisted after Bonferroni correction. However, no causal effect of LNSP on cervical cancer was found according to the IVW results (OR = 1.003, 95 % CI: 1.000, 1.007, P = 0.071). From the results of MR-PRESSO and MR-Egger, no SNP with horizontal pleiotropy between cervical cancer was detected and no SNP was identified as an outlier SNP. Cochran's Q statistic suggested that no heterogeneity existed among IVs of AFSI and LNSP. According to Leave-one-out analysis, the results of MR did not change after excluding any single IV. Conclusion: This MR study reveals that early AFSI has a causal effect on cervical cancer.

2.
Psychiatry Clin Psychopharmacol ; 32(3): 205-214, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-38766673

RESUMO

Background: It has been shown that autoimmune diseases are associated with psychiatric disorders in epidemiological studies. The acute psychiatric disorder patients have higher frequency of autoantibodies in the blood, including antinuclear antibodies, anti-thyroid peroxidase, and thyroglobulin [thyroid antibody carriers]. However, large clinical studies with more relevant control groups in China are few. Methods: This was a retrospective study. A total of 1669 sera were tested for autoantibodies in the clinical laboratory of the Fourth Affiliated Hospital, Zhejiang University School of Medicine from October 2016 to March 2021. All data available during this time period were analyzed. Only the first entry for each patient from inpatient care units was used for analysis. The clinical information and laboratory data of patients were retrospectively collected and analyzed. Results: A significantly lower prevalence of antinuclear antibodies was observed in the healthy control group than in the patient group (21.7% vs 28.8%, P < .05). There was a significant difference in the prevalence of antinuclear antibodies between thyroglobulin-antibody carriers and thyroid peroxidase-antibody- and thyroglobulin-antibody-seronegative individuals in the unipolar depressive disorder group (P < .05). A positive anti-thyroid peroxidase test was significantly associated with patients having nonaffective psychoses (P < .05). Conclusion: The results showed that psychiatric disorders were associated with antinuclear antibodies and thyroid autoantibodies in our large sample of patients admitted to acute psychiatric hospitalization, and autoimmune autoantibodies were potential biomarkers of psychotic disorders. The results might lead to new research directions for the study of psychiatric disorders in the future.

3.
Nat Commun ; 12(1): 3533, 2021 Jun 10.
Artigo em Inglês | MEDLINE | ID: mdl-34112798

RESUMO

In subduction zones, materials on Earth's surface can be transported to the deep crust or mantle, but the exact mechanisms and the nature of the recycled materials are not fully understood. Here, we report a set of migmatites from western Yangtze Block, China. These migmatites have similar bulk compositions as forearc sediments. Zircon age distributions and Hf-O isotopes indicate that the precursors of the sediments were predominantly derived from juvenile arc crust itself. Using phase equilibria modeling, we show that the sediments experienced high temperature-to-pressure ratio metamorphism and were most likely transported to deep arc crust by intracrustal thrust faults. By dating the magmatic zircon cores and overgrowth rims, we find that the entire rock cycle, from arc magmatism, to weathering at the surface, then to burial and remelting in the deep crust, took place within ~10 Myr. Our findings highlight thrust faults as an efficient recycling channel in compressional arcs and endogenic recycling as an important mechanism driving internal redistribution and differentiation of arc crust.

4.
BMC Infect Dis ; 20(1): 329, 2020 May 07.
Artigo em Inglês | MEDLINE | ID: mdl-32381073

RESUMO

BACKGROUND: Although people of all ages are susceptible to the novel coronavirus infection, which is presently named "Coronavirus Disease 2019" (COVID-19), there has been relatively few cases reported among children. Therefore, it is necessary to understand the clinical characteristics of COVID-19 in children and the differences from adults. CASE PRESENTATION: We report one pediatric case of COVID-19. A 14-month-old boy was admitted to the hospital with a symptom of fever, and was diagnosed with a mild form of COVID-19. The child's mother and grandmother also tested positive for SARS-CoV-2 RNA. However, the lymphocyte counts were normal. The chest computed tomography (CT) revealed scattered ground glass opacities in the right lower lobe close to the pleura and resorption after the treatment. The patient continued to test positive for SARS-CoV-2 RNA in the nasopharyngeal swabs and stool at 17 days after the disappearance of symptoms. CONCLUSION: The present pediatric case of COVID-19 was acquired through household transmission, and the symptoms were mild. Lymphocyte counts did not significantly decrease. The RNA of SARS-CoV-2 in stool and nasopharyngeal swabs remained positive for an extended period of time after the disappearance of symptoms. This suggests that attention should be given to the potential contagiousness of pediatric COVID-19 cases after clinical recovery.


Assuntos
Infecções por Coronavirus/diagnóstico , Coronavirus , Fezes/virologia , Febre/etiologia , Pulmão/diagnóstico por imagem , Nasofaringe/virologia , Pneumonia Viral/diagnóstico por imagem , Adulto , Betacoronavirus , COVID-19 , Teste para COVID-19 , Técnicas de Laboratório Clínico , Coronavirus/genética , Coronavirus/isolamento & purificação , Infecções por Coronavirus/diagnóstico por imagem , Infecções por Coronavirus/epidemiologia , Características da Família , Humanos , Lactente , Contagem de Linfócitos , Masculino , Pandemias , Pneumonia Viral/epidemiologia , Reação em Cadeia da Polimerase , SARS-CoV-2 , Síndrome Respiratória Aguda Grave/transmissão , Tomografia Computadorizada por Raios X
5.
Rapid Commun Mass Spectrom ; 33(4): 361-370, 2019 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-30450611

RESUMO

RATIONALE: In situ trace element analysis of melt inclusions by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) provides important geochemistry information. However, the precision and accuracy of this technique are affected by many factors, such as matrix effect, laser conditions, and calibration method. In addition, many previous LA-ICP-MS studies ablated entire melt inclusions along with their host minerals and obtained trace element composition by deconvoluting the mixed ablation signal, which may induce much uncertainty. METHODS: A 193 nm ArF laser ablation system combined with inductively coupled plasma sector field mass spectrometry (ICP-SF-MS) was used to investigate matrix effect, laser conditions, choice of external calibration standards, and data reduction strategy for in situ analysis of 36 major and trace elements in six common silicate reference glasses. The validity of the protocol presented here was demonstrated by measuring trace elements in olivine-hosted melt inclusions. Instead of ablating entire melt inclusions along with their host minerals, melt inclusions were polished to the surface to avoid laser ablating the mineral host. RESULTS: The calibration lines calculated from the calibration standards should cross the coordinate origin, especially for low-concentration elements (<10 ppm). As the laser crater size increased from 17 to 33 µm, the precision was improved from <20% to <8% (2RSD), and accuracy was improved from ±20% to better than ±10%. Most measured trace elements in Dali melt inclusions are consistent with those in their host rocks. For mobile elements (Ba, Sr, Pb), melt inclusions display much smaller variations than their host rocks. CONCLUSIONS: A simple but accurate approach for in situ analysis of trace elements in melt inclusions by LA-ICP-SF-MS has been established, which should greatly facilitate the wider application of in situ trace element geochemistry to melt inclusion studies.

6.
Int J Ophthalmol ; 11(12): 1978-1983, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30588433

RESUMO

AIM: To investigate the associations between demographic and clinical factors with the rate of visual field mean derivation (MD) decline in retinitis pigmentosa (RP) patients. METHODS: Correlations of MDs with the visual acuity and retinal pigmentation were analyzed in 928 RP patients. MD decreasing rate in 10y and potential influences of gender, age, family history and retinal pigmentation on the rate were explored in 201 RP patients. RESULTS: In the 928 patients, average MD and visual acuity were -14.44±8.61 dB and 0.79±0.35 respectively and when MD was lower than -9.18 dB the visual acuity would be below 1.0 (20/20). The average MD medium between eyes with or without retinal pigmentation was -14.82 dB. In 123 non-pigmented eyes, the average MD were lower than the medium but in 153 pigmented eyes it was higher than that. In the 201 patients, the average decreasing value of MD in 10 years' period was -8.01±3.66 dB and the value were correlated to retinal pigmentation but not to gender, age or RP family history. CONCLUSION: The rate of MD decline in RP eyes is significantly related to retinal pigmentation. Our study demonstrates the quantitative rate of MD decline in RP patients and the value of MD could well reflect the severity of RP.

7.
Zhongguo Zhong Yao Za Zhi ; 41(23): 4428-4435, 2016 Dec.
Artigo em Chinês | MEDLINE | ID: mdl-28933123

RESUMO

To systematically review the clinical efficacy and safety of Yinzhihuang oral liquid in the treatment of intrahepatic cholestasis of pregnancy(ICP). Literatures published by June 2016 were searched in databases, such as Medline, Pubmed, Cochrane Library, China National Knowledge Infrastructure(CNKI), Chinese Scientific Journals Full-text Database(VIP), Chinese biomedical literature database(CBM), and Wanfang Database. Randomized controlled trials(RCT) of Yinzhihuang oral liquid were collected according to the inclusion criteria, and the methodological quality of selected literatures was evaluated. The Meta-analysis was conducted by using RevMan 5.3 software. A total of 7 RCTs involving 711 patients were included. The results of Meta-analysis showed that, compared with control group, Yinzhihuang oral liquid significantly alleviated pruritus symptoms[MD=-0.68, 95%CI(-0.95,-041), P<0.000 01], reduced blood biochemical indexes including TBA[MD=-7.23, 95%CI (-10.88,-3.58), P=0.000 1], TB[MD=-1.90, 95%CI(-3.09,-0.70), P=0.002], ALT[MD=-39.08, 95%CI (-56.46,-21.70), P<0.000 1], and CG [MD=-0.71, 95%CI(-0.89,-0.52), P<0.000 01]. In the respect of pregnancy outcome, Yinzhihuang oral liquid can distinctly improve birth weight [MD=430.03, 95%CI (219.28, 640.78), P<0.000 1]. However, there was no significant difference in cesarean section rate [OR=0.93, 95%CI (0.36, 2.36), P=0.87], preterm birth rates [OR=0.63, 95%CI (0.28, 1.42), P=0.26], and neonatal asphyxia rate [OR=0.50, 95%CI (0.18, 1.43), P=0.20]. Yinzhihuang oral liquid showed better efficiency and slighter adverse reaction. However, more rigorously designed, double-blind, randomized controlled trials with large sample size and high quality are required to provide further evidences.


Assuntos
Colestase Intra-Hepática/tratamento farmacológico , Medicamentos de Ervas Chinesas/uso terapêutico , Complicações na Gravidez/tratamento farmacológico , China , Feminino , Humanos , Gravidez , Ensaios Clínicos Controlados Aleatórios como Assunto
10.
Hypertens Res ; 35(12): 1129-34, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22914556

RESUMO

A number of studies have investigated the association between the methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism and the risk of pre-eclampsia (PE) in various populations and have delivered inconsistent results. Therefore, this meta-analysis of 36 case-control studies, comprising 4253 PE cases and 4950 controls, were assessed to evaluate a possible association. The pooled results showed that the MTHFR C677T polymorphism was significantly associated with PE (P=0.03, odds ratio (OR)=1.25, 95% confidence interval (CI)=1.02-1.54, for the additive comparison; P=0.04, OR=1.14, 95% CI=1.01-1.29, for the dominant genetic model). The results of the subgroup analysis showed that MTHFR 677T had the effect of increasing the PE risk for the recessive genetic model (P<0.0001, OR=1.76, 95% CI=1.33-2.33, P(heterogeneity)=0.28), the additive comparison (P=0.002, OR=2.09, 95% CI=1.31-3.31, P(heterogeneity)=0.08) and allele contrasts (P=0.03, OR=1.42, 95% CI=1.04-1.95, P(heterogeneity)=0.0001) in the Asians, while no evidence of an association between MTHFR C677T polymorphisms and PE was observed in the Caucasians. This meta-analysis suggests that the MTHFR C677T polymorphism is capable of causing PE susceptibility in the Asians but not in the Caucasians.


Assuntos
Predisposição Genética para Doença , Metilenotetra-Hidrofolato Redutase (NADPH2)/genética , Polimorfismo Genético , Pré-Eclâmpsia/genética , Feminino , Humanos , Pré-Eclâmpsia/etiologia , Gravidez , Viés de Publicação
11.
Guang Pu Xue Yu Guang Pu Fen Xi ; 29(4): 1104-7, 2009 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-19626912

RESUMO

Deficiency of micronutrients, especially iron and zinc, has been a serious malnutrition problem worldwide in human health. Increasing Fe and Zn concentrations in grains by means of plant breeding is a sustainable, effective and important way to improve human mineral nutrition and health. However, little information on grain Fe and Zn concentrations in Chinese wheat genotypes is available. Therefore, to determine the nutrients status especially these of micronutrients in wheat grain is necessary and very useful. Two hundred sixty two genotypes were selected from the wheat mini-core collections, which contained 23090 wheat genotypes in China and represented 72.2% of total genetic variation. All 262 genotypes were grown in soils of similar geographical and climate location in order to minimize the environmental effect. After harvesting, the grains were washed with deionized water and dried (around 70 degrees C), then digested in HNO3 solution using a microwave accelerating reaction system (MARS). Nutrient concentrations in stock solution were analyzed by inductively coupled plasma atomic emission spectrometry (ICP-AES). Remarkable genetic variations among grain nutrient concentrations (Fe, Mn, Cu, Zn, Mg, Ca, K and P ) in the tested genotypes were detected. The concentrations of Fe, Zn, Mn, Cu, Ca, Mg, K and P in wheat grain were in the ranges of 34.2-61.2, 26.3-76.0, 20.9-56.7, 3.4-9.8, 290-976, 1129-2210 mg x kg(-1); 0.34%-0.85% and 0.296%-0.580%, respectively. The corresponding average values were 45.1, 50.2, 37.9, 6.5, 515, 1772 mg x kg(-1), 0.55% and 0.451%, respectively. Significant positive correlations between micronutrients (Fe, Mn, Zn, and Cu) in wheat grains were detected, and the correlation coefficients were 0.395** (Fe and Mn), 0.424** (Fe and Zn), 0.574** (Fe and Cu), and 0.474** (Mn and Cu), respectively. However, no significant difference was found in grain nutrient concentrations between spring-wheat and winter-wheat genotypes. This study provides valuable and important information for breeding wheat genotypes which are enriched with minerals in grains, especially Fe and Zn


Assuntos
Extratos Vegetais/análise , Espectrofotometria Atômica/métodos , Oligoelementos/análise , Triticum/química , China , Valor Nutritivo
12.
Zhejiang Da Xue Xue Bao Yi Xue Ban ; 38(3): 276-82, 2009 05.
Artigo em Chinês | MEDLINE | ID: mdl-19504637

RESUMO

OBJECTIVE: To construct prokaryotic expression systems of TCS genes comD/comE/comC of Streptococcus pneumoniae, and to determine the correlation of ComD and ComC with the drug resistance. METHODS: The entire comD, comE and comC genes were amplified by PCR and their prokaryotic expression systems were established by routine genetic engineering technique. SDS-PAGE and Bio-Rad Agarose Image Analyzor was applied to measure the outputs of target recombinant proteins rComD, rComE and rComC. Rabbits were immunized with these recombinant proteins to prepare antisera. The resistance of S.pneumoniae strains to penicillin and cefotaxime was examined after ComD and ComC were blocked by antisera. RESULT: Compared with the reported sequences, similarities of nucleotide and amino acid sequences of the cloned comD, comE and comC genes were 98.4% approximately 99.3% and 99.1% approximately 100%, respectively. The constructed engineering bacteria E.coli BL21DE3(pET42a-comD), E.coli BL21DE3(pET42a-comE) and E.coli BL21DE3(pET42a-comC) were able to efficiently express the target recombinant proteins and the outputs of rComD, rComE and rComC were 28%, 25% and 35% of the total bacterial proteins, respectively. The double immunodiffusion titers of rabbit antisera against rComD, rComE or rComC were 1:4, 1:4 and 1:8, respectively. After the ComD and/or ComC were blocked by the antisera, the cefotaxime-sensitive S. pneumoniae strains became to resistant to antibiotics but there were no changes for cefotaxime-resistant strains and resistance to penicillin for all tested strains. CONCLUSION: The prokaryotic expression systems of S.pneumoniae comD/come/comC genes have been successfully constructed, and the study also indicates that both the ComD and ComC are involved in the drug resistance of S. pneumoniae to cefotaxime.


Assuntos
Proteínas de Bactérias/genética , Streptococcus pneumoniae/efeitos dos fármacos , Streptococcus pneumoniae/genética , Resistência beta-Lactâmica/genética , Animais , Proteínas de Bactérias/biossíntese , Cefotaxima/farmacologia , Escherichia coli/genética , Escherichia coli/metabolismo , Coelhos , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/genética , Recombinação Genética , Transdução de Sinais
13.
Zhejiang Da Xue Xue Bao Yi Xue Ban ; 37(6): 605-11, 2008 11.
Artigo em Chinês | MEDLINE | ID: mdl-19084959

RESUMO

OBJECTIVE: To construct prokaryotic expression systems of Streptococcus pneumoniae ciaH and ciaR genes,and to determine their correlation with drug resistance. METHODS: The total length of ciaH and ciaR genes was amplified by PCR and their prokaryotic expression systems were established by using routine genetic engineering technique. SDS-PAGE was applied to measure the outputs of target recombinant proteins rCiaH and rCiaR. Rabbits antisera and IgGs against rCiaH and rCiaR were prepared. The resistance to penicillin and cefotaxime of S.pneumoniae strains was examined after CiaH and CiaR were extracellularly and intracellularly blocked by the IgGs. RESULT: The homogeneity of nucleotide and amino acid sequences of the cloned ciaH and ciaR genes with the reported sequences was 99.9-100% and 100%, respectively. The recombinant bacteria E.coli BL21DE3pET42a-ciaH and E.coli BL21DE3pET42a-ciaR were able to express the target recombinant proteins rCiaH and rCiaR with efficiency. The outputs of rCiaH and rCiaR were 33% and 45% of the total bacterial proteins, respectively. The double immunodiffusion titers of rCiaH antiserum,rCiaR antiserum,rCiaH-IgG and rCiaR-IgG were 1:4,1:4,1:1 and 1:1, respectively. After CiaH was extracellularly or intracellularly blocked by CiaH-IgG, and CiaR was intracellularly blocked by CiaR-IgG, the penicillin-sensitive or cefotaxime-sensitive strains developed resistance to the two antibiotics; but the blocks did not change that of penicillin-resisting or cefotaxime-resisting strains. CONCLUSION: The prokaryotic expression systems of S. pneumoniae ciaH/ciaR genes have been successfully constructed in this study. Both the CiaH and CiaR may be involved in penicillin and cefotaxime resistance of the bacterium.


Assuntos
Proteínas de Bactérias/metabolismo , Proteínas Quinases/metabolismo , Streptococcus pneumoniae/efeitos dos fármacos , Streptococcus pneumoniae/genética , Resistência beta-Lactâmica/genética , Animais , Proteínas de Bactérias/genética , Cefotaxima/farmacologia , Escherichia coli/genética , Escherichia coli/metabolismo , Regulação Bacteriana da Expressão Gênica , Vetores Genéticos , Resistência às Penicilinas/genética , Proteínas Quinases/genética , Coelhos , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Transdução de Sinais , Streptococcus pneumoniae/enzimologia
14.
Vaccine ; 26(51): 6639-44, 2008 Dec 02.
Artigo em Inglês | MEDLINE | ID: mdl-18834913

RESUMO

The Vi capsular polysaccharide vaccine is one of two vaccines against typhoid recommended worldwide and is the vaccine generally used in China. However, in recent years a Salmonella paratyphi A strain that is naturally devoid of capsule has caused frequent outbreaks of typhoid fever in Southern China, leading to the need for identification of additional antigens that could be incorporated into new vaccines. SpaO acts as a major invasion factor of Salmonella enterica spp. and H1a is the unique flagellin subunit ofS. paratyphi A. In this study, the two prokaryotic recombinant antigens, rSpaO and rH1a, were expressed and their immunogenicity was demonstrated by the slide agglutination test and Western blot assays. Using PCR and sequencing analysis as well as ELISA, we find that the spaO and h1a genes are widely distributed in 196 S. paratyphi A isolates (97.5 and 100%, respectively), with high expression frequencies for the SpaO (98.0%) and H1a (100%) antigens. The two genes also show high sequence conservation (similarities from 99.31 to 99.88% for both genes). In sera from 172 paratyphoid A patients, anti-SpaO and anti-H1a IgGs were detectable by ELISA, in 94.8 and 98.8% of patients, respectively. Furthermore, 41.7-66.7% of mice immunized with rSpaO or rH1a alone were protected against subsequent infection, and the protection rate rose to 75.0-91.7% in mice co-immunized with the two antigens. As the spaO and h1a genes of S. paratyphi A are sequence conserved, extensively distributed and highly expressed, the rSpaO and rH1a immunogens should be considered in the development of novel vaccines to prevent S. paratyphi A-caused typhoid fever.


Assuntos
Proteínas de Bactérias/imunologia , Proteínas de Membrana/imunologia , Febre Paratifoide/prevenção & controle , Salmonella paratyphi A/imunologia , Vacinas Tíficas-Paratíficas/imunologia , Animais , Anticorpos Antibacterianos/sangue , Anticorpos Antibacterianos/imunologia , Antígenos de Bactérias/genética , Antígenos de Bactérias/imunologia , Proteínas de Bactérias/genética , DNA Bacteriano/genética , Humanos , Proteínas de Membrana/genética , Camundongos , Camundongos Endogâmicos BALB C , Febre Paratifoide/imunologia , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Salmonella paratyphi A/genética , Análise de Sequência de DNA
15.
Zhonghua Liu Xing Bing Xue Za Zhi ; 26(2): 120-3, 2005 Feb.
Artigo em Chinês | MEDLINE | ID: mdl-15921614

RESUMO

OBJECTIVE: To establish a 16S rDNA multiplex polymerase chain reaction (PCR) for simultaneously detecting P. gingivalis, A. actinomycetemcomitans and T. denticola in clinical specimens of chronic periodontitis and to study the correlation between different modes of infection and severity of the disease. METHODS: Periodontal pocket specimens from 152 patients with mild, moderate or advanced chronic periodontitis and gingival sulcus specimens from 30 periodontally healthy individuals were collected and placed in 200 microl lysis buffer. The specimens were incubated in 100 degrees C for 10 min and 10 microl of the supernatant was directly used as PCR template. DNAs from P. gingivalis strain ATCC33277, A. actinomycetemcomitans strain Y4, T. denticola strain FM and E. coli strain DH5alpha were used as positive and negative controls in PCR with all of which were prepared by routing phenol-chloroform method. A multiplex PCR assay, using three sets of primers specific to 16S rDNA genes of the three anaerobes, was developed to detect the specimens. The target amplification fragments from 3 cases of PCR products positive for all the three anaerobes were sequenced after T-A cloning. Chi-square test was applied to analyze the correlation between different coinfection of the three anaerobes and severity of the disease. RESULTS: The established 16S rDNA multiplex PCR assay was able to detect P. gingivalis, A. actinomycetemcomitans and T. denticola at a minimum of 10, 20 and 20 cells, respectively. In comparison with the reported corresponding sequences, similarities of the nucleotide sequences from the three anaerobes 16S rDNA amplification fragments were as high as 99.45%, 97.08% and 96.59%, respectively. Of the 30 periodontally healthy gingival sulcus specimens, only one (3.3%) positive for P. gingivalis and two (6.7%) for A. actinomycetemcomitans could be identified but all of the rest were negative. In the 152 CP periodontal pocket specimens, 147 cases (96.7%) were positive for P. gingivalis, A. actinomycetemcomitans and/or T. denticola, respectively, and 5 cases (3.3%) were negative for all the three anaerobes. The positive rate of P. gingivalis detection (91.5%, 139/152) was significantly higher than those of A. actinomycetemcomitans (72.4%, 110/152) and T. denticola (80.9%, 123/152) (chi(2) = 7.07, 18.67; P < 0.01). 89.8% of the specimens from patients showed coinfections with two (26.5%) or three anaerobes (63.3%), and the coinfection rates in the specimens from moderate and advanced CP were remarkably higher than that from mild CP (chi(2) = 10.43, P < 0.01). CONCLUSION: The 16S rDNA multiplex PCR established in this study showed high sensitivity and specificity, which could be used to detect P. gingivalis, A. actinomycetemcomitans and T. denticola in clinical specimens. CP was an disease caused by multiple pathogenic microbes while the synergistic pathopoiesis of the three microbes was closely related to the severity of the disease.


Assuntos
Aggregatibacter actinomycetemcomitans/isolamento & purificação , Periodontite/microbiologia , Porphyromonas gingivalis/isolamento & purificação , Treponema denticola/isolamento & purificação , Infecções por Actinobacillus/epidemiologia , Infecções por Actinobacillus/microbiologia , Adulto , Idoso , Infecções por Bacteroidaceae/epidemiologia , Infecções por Bacteroidaceae/microbiologia , China/epidemiologia , Doença Crônica , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase/métodos , RNA Ribossômico 16S , Infecções por Treponema/microbiologia
16.
Zhejiang Da Xue Xue Bao Yi Xue Ban ; 32(1): 17-20, 2003 02.
Artigo em Chinês | MEDLINE | ID: mdl-12640703

RESUMO

OBJECTIVE: To clone the LTB gene of E.coli and the CTB gene of V.cholerae, and to construct expression vectors of these genes. METHODS: The LTB gene from E.coli strain 44815 and the CTB gene from V.cholerae strain eastern 74 were amplified by high fidelity PCR. The nucleotide sequences of the two target DNA amplification fragments were sequenced after T-A cloning. pET32a expression vectors with inserted LTB and CTB genes were constructed. The LTB and CTB fusion proteins were expressed in E.coli strain BL21DE3 inducted by IPTG at different dosages. The two expression products were identified by SDS-PAGE and G(M1)-ELISA. RESULTS: In comparison with the reported LTB and CTB sequences, the nucleotide sequence homologies of the cloned LTB gene and CTB gene were from 99.12% approximate, equals 99.71% and 98.54% approximate, equals 99.42%, while their putative amino acid sequence homologies were as high as 97.58% approximate, equals 99.19% and 96.77% approximate, equals 99.19%. The expression outputs of LTB and CTB fusion proteins in pET32a LTB BL21DE3 and pET32a-CTB-BL21DE3 systems were approximately 30% and 10% of the total bacterial proteins, respectively. The LTB and CTB fusion proteins were able to combine with bovine G(M1) confirmed by ELISA. CONCLUSION: The expression systems of LTB and CTB genes have been successfully established. Both the expressed LTB and CTB fusion proteins possess mucosal adjuvant immunoactivity.


Assuntos
Adjuvantes Imunológicos/farmacologia , Toxinas Bacterianas/genética , Toxina da Cólera/genética , Enterotoxinas/genética , Proteínas de Escherichia coli , Escherichia coli/genética , Vibrio cholerae/genética , Animais , Toxinas Bacterianas/farmacologia , Sequência de Bases , Toxina da Cólera/farmacologia , Clonagem Molecular , Enterotoxinas/farmacologia , Imunidade nas Mucosas , Coelhos
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