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The meta-analysis aimed to systematically evaluate all the available pieces of evidence concerning the clinical effectiveness of Er,Cr:YSGG lasers (erbium, chromium, yttrium scandium gallium garnet laser) in the non-surgical treatment of patients with chronic periodontitis, and provide guidance for clinicians about the application of Er,Cr:YSGG lasers during the process of non-surgical periodontal treatments. The meta-analysis was conducted with data extracted from 16 randomized controlled clinical trials (RCTs) that compare Er,Cr:YSGG lasers adjunct/substitute to scaling and root planing (SRP) with SRP alone for the treatment of chronic periodontitis published in English or Chinese from January 2000 to January 2020. The weighted mean differences (WMDs) and 95% confidence intervals (CIs) were counted for probing depth (PD) reduction, clinical attachment level (CAL) gain, and visual analogue scale (VAS) score. Heterogeneity of each study was evaluated with the Q test. The publication bias was measured using Begg's adjusted rank correlation test. Sixteen RCTs with 606 patients were included in the meta-analysis. There were statistically significant differences between Er,Cr:YSGG lasers adjunct/substitute to SRP and SRP alone in the PD reduction at 1-month follow-up (WMD = 0.35, 95% CI [- 0.63, 0.07], P = 0.013), 3-month follow-up (WMD = - 0.342, 95% CI [- 0.552, - 0.132], P = 0.001), CAL gain at 3-month follow-up (WMD = - 0.17, 95% CI [- 0.31, 0.03], P = 0.017), and VAS score (WMD = - 2.395, 95% CI [- 3.327, - 1.464], P = 0.000) immediately after treatment. There were no significant differences of PD reduction and CAL change at 6-month follow-up. The present meta-analysis indicated that Er,Cr:YSGG lasers provided additional effectiveness in PD reduction and CAL gain at short-term follow-ups and there was less pain compared with SRP alone.
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Periodontite Crônica/radioterapia , Lasers de Estado Sólido/uso terapêutico , Ensaios Clínicos Controlados Aleatórios como Assunto , Humanos , Resultado do TratamentoRESUMO
Spexin (Spx), a novel neuropeptide, composed of 14 amino acid residues, is evolutionally conserved from fish to mammals. It has been suggested that Spx has pleiotropic functions in mammals. However, reports about Spx are very limited. To clarify the roles of Spx in the regulation of reproduction and food-intake in the spotted scat, the spx (ssspx) gene was cloned and analyzed. Analysis of the tissue distribution by RT-PCR showed that ssspx expression was widespread. During ovary development, expression of ssspx was found to be highest in phase II, moderate in phase III, and at its lowest level in phase IV. Ssspx expression was significantly down-regulated in the hypothalamus after treatment with E2 both in vitro and in vivo. A significant increase of ssspx was observed after 2 and 7â¯days of food deprivation. However, the ssspx transcript levels in the 7â¯day fasting group decreased significantly after refeeding 3â¯h after the scheduled feeding time. This suggests that ssSpx may be involved in the regulation of reproduction and food-intake in the spotted scat.
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Perfilação da Expressão Gênica , Hormônios Peptídicos/genética , Perciformes/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , Estradiol/farmacologia , Jejum , Feminino , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Hipotálamo/efeitos dos fármacos , Hipotálamo/metabolismo , Masculino , Ovário/efeitos dos fármacos , Ovário/embriologia , Ovário/metabolismo , Hormônios Peptídicos/química , Hormônios Peptídicos/metabolismo , Perciformes/metabolismo , Filogenia , Reprodução , Alinhamento de Sequência , Distribuição Tecidual/efeitos dos fármacosRESUMO
OBJECTIVE: To explore the clinical characteristics of pregnant uterine rupture, enhance its early diagnosis rate and improve its prognosis. METHODS: For this retrospective study, a total of 55 143 delivery cases at Tongzhou District Maternal & Child Health Hospital from January 1, 2003 to May 31, 2012 was analyzed. There were 2490 cases of scarred uterus and 68 cases had uterine rupture. And 62 of them belonged to scarred uterus rupture. And there were 52 653 cases of non-scarred uterus childbirth and 6 of them had rupture. RESULTS: Significant differences existed in the rupture of scarred versus non-scarred uterus and maternal versus primipara women. Uterus rupture usually occurred at the incomplete rupture position of original surgical scar at 39 weeks or more. The scar thickness of lower uterine segment was ≤ 3 mm on ultrasound. Three cases with laparoscopic myoma removal within 1 year had complete rupture at 36-38 weeks. Postpartum hemorrhage incidence was 42.65%. There were two cases of hysterectomy and 11 cases of neonatal asphyxia. CONCLUSION: For second pregnancy of cesarean section, pregnancy should be terminated before 39 weeks if scar thickness of lower uterine segment is ≤ 3 mm. Late rupture is to be monitored closely for those with laparoscopic myoma removal within 1 year. Meanwhile, we should strengthen the management of migrant and unemployed women.
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Ruptura Uterina , Adulto , Cesárea/efeitos adversos , Feminino , Humanos , Gravidez , Estudos Retrospectivos , Ruptura Uterina/epidemiologia , Ruptura Uterina/etiologia , Ruptura Uterina/prevenção & controle , Adulto JovemRESUMO
Tetrahydropalmatine (THP) is an active component of Corydalis yanhusuo W. T. Wang. The current study investigates the possible cardioprotective effects of tetrahydropalmatine in acute myocardial ischemia (AMI) rats. The anterior descending coronary artery of SD rats was ligated to establish an AMI model. After two weeks of gavage of THP, cardiac function was determined by echocardiography. The organ index and the infarct size were assessed after the experiment, and the histopathological myocardial tissue changes were observed. In addition, the apoptosis index of myocardial cells was detected by the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. The levels of SOD, MDA, CAT, GSH-Px, BNP, and cTn-I were measured by enzyme-linked immunosorbent assay. To determine relevant proteins, the Western blot and molecular docking were applied. Compared with the model group, THP could enhance rat cardiac ejection function to improve cardiac function, drastically lessen the infarct size, reduce myocardial cell damage and inflammatory cell infiltration. THP might also prevent ischemic myocardial damage by inhibiting myocardial cell apoptosis and efficiently reducing oxidative stress. Specifically, THP could decrease MDA, BNP, c-TnI activities, as well as the expression of Bax and Caspase-3 protein, while increasing SOD, GSH-Px, CAT activities, and Bcl-2 level. Furthermore, THP could significantly promote the phosphorylation of PI3K and Akt proteins. The involved pathways and proteins have also been verified through molecular docking. According to these findings, THP may preserve the myocardium due to its anti-oxidant and anti-apoptotic properties.
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Infarto do Miocárdio , Isquemia Miocárdica , Ratos , Animais , Simulação de Acoplamento Molecular , Ratos Sprague-Dawley , Infarto do Miocárdio/tratamento farmacológico , Infarto do Miocárdio/metabolismo , Infarto do Miocárdio/patologia , Miocárdio/patologia , Miócitos Cardíacos/patologia , Apoptose , Isquemia Miocárdica/patologia , Superóxido Dismutase/metabolismoRESUMO
Muscone is the main active compound of Moschus. In this paper, the cardioprotective effect of Muscone on acute myocardial ischemia (AMI) rats and its potential mechanisms were investigated. AMI rat models were established to evaluate the protective effect and antioxidative function of Muscone on the hearts. Moreover, Western blot analysis was conducted to quantify the phosphorylated PI3K and AKT levels in PI3K/Akt pathway for further investigating the mechanism of Muscone. Results showed that Muscone could markedly lessen the infarct size and myocardial injury, improve cardiac function, inhibit cardiomyocyte apoptosis and down-regulate serum reactive oxygen species level as indicated by the decreased MDA, BNP and c-TnI activities and the increased SOD, GSH-px, CAT activities and the expression of Bax protein. In addition, it was revealed that Muscone notably promoted the phosphorylation of PI3K and AKT. These findings denote that Muscone exerts a protective effect in heart via inhibition of oxidative stress and apoptosis, offering new insights into the treatment of CHD and the clinical application of Muscone.
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Cicloparafinas , Isquemia Miocárdica , Transdução de Sinais , Animais , Cicloparafinas/farmacologia , Isquemia Miocárdica/tratamento farmacológico , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Ratos , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/metabolismo , Proteína X Associada a bcl-2/metabolismo , Proteína X Associada a bcl-2/farmacologiaRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Genkwa flos, as a traditional herb, is the dried flower buds of Daphne genkwa Sieb.et Zucc. It is used in traditional medicine for the treatment of cough, sore throats, edema. AIM OF THE STUDY: The study aimed to explore a new mathematical method for multivariate evaluation, investigate the anti-inflammatory and anti-angiogenic activities of flavonoids in Daphne Genkwa under ex vivo conditions. MATERIALS AND METHODS: The flavonoids monomers in Daphne Genkwa were separated by preparative liquid chromatography and identified by HPLC-ESI-ITMS. An in vitro inflammatory model of macrophage RAW264.7 induced by LPS and an angiogenesis model of human umbilical vein endothelial cells induced by TNF-α were established. Flavonoids were extracted and prepared for intervention to detect the amount of secretion after drug intervention to reflect the anti-inflammatory and anti-angiogenic activities of each component. In addition, a new mathematical method, which combined principal component analysis and efficacy coefficient method, was adopted in pharmacodynamic evaluation. RESULTS: Fourteen flavonoids monomers were separated by preparative liquid chromatography and identified by HPLC-ESI-ITMS including H1 (hydroxygenkwanin-5-O-ß-D-glucoside), H2 (apigenin-7-O-ß-D-glucoside), H3 (kaempferol-3-O-ß-D-glucoside), H4 (hydroxygenkwanin-5-O-ß-D-primeveroside), H5 (apigenin-5-O-ß-D-primeveroside), H6 (apigenin-7-O-ß-D-glucuronide), H7 (luteolin-5-O-ß-D-glucopyranoside), H8 (genkwain-5-O-ß-D- glucoside), H9 (luteolin), H10 (Daphnodorin G), H11 (tiliroside), H12 (apigenin), H13 (3'- hydroxygenkwain) and H14 (genkwanin). We found that most of flavonoids down-regulated VCAM and MMP-3, while H1, H8, H9, H14 reduced VEGF and ICAM was only decreased by H14. CONCLUSION: Genkwanin may be the most active anti-rheumatoid arthritis flavonoids in Daphne genkwa. Meanwhile, the new mathematical method used in the study provided a new direction for solving the problem of multi-index pharmacodynamic evaluation.
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Inibidores da Angiogênese/farmacologia , Anti-Inflamatórios/farmacologia , Daphne , Flavonoides/farmacologia , Extratos Vegetais/farmacologia , Análise de Componente Principal/métodos , Inibidores da Angiogênese/isolamento & purificação , Animais , Anti-Inflamatórios/isolamento & purificação , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/fisiologia , Avaliação Pré-Clínica de Medicamentos/métodos , Quimioterapia Combinada , Flavonoides/isolamento & purificação , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Camundongos , Extratos Vegetais/isolamento & purificação , Células RAW 264.7 , RatosRESUMO
SheXiang XinTongNing (XTN), which is composed of six traditional Chinese herbs, is a commercially available Chinese patent medicine that has been widely used as the treatment of coronary heart disease (CHD). Its mechanisms against coronary heart disease, however, remain largely unknown. This study aimed to investigate the pharmacological mechanisms of XTN against CHD via network pharmacology and experimental evaluation. In this study, GO enrichment and KEGG pathway enrichment were firstly performed for acquiring the potentially active constituents of XTN, the candidate targets related to coronary heart disease, the drug-components-targets network as well as the protein-protein interaction network and further predicting the mechanisms of XTN against coronary heart disease. Subsequently, a series of in vitro experiments, specifically MTT assay, flow cytometry and Real-time quantitative polymerase chain reaction analysis, and a succession of in vivo experiments, including Tunel staining and immunohistochemical staining were conducted for further verification. Results showed that Bcl-2, IGF1, CASP3 were the key candidate targets which significantly associated with multiple pathways, namely PI3K-Akt signaling pathway and MAPK signaling pathway. It indicated that the potential mechanism of XTN against CHD may be predominantly associated with cell apoptosis. The in vitro experimental results showed that XTN treatment remarkably decreased the apoptotic rate and Bax/Bcl-2 ratio of H9c2 cells. Histological results confirmed that XTN not only effectively alleviated oxidative damage caused by myocardial ischemia but inhibited cell apoptosis. Given the above, through the combined utilization of virtual screening and experimental verification, the findings suggest that XTN makes a significant contribution in protecting the heart from oxidative stress via regulating apoptosis pathways, which lays the foundations and offers an innovative idea for future research.
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ETHNOPHARMACOLOGICAL RELEVANCE: Duijinsan (DJS) is a famous Chinese medicine prescription composed of Radix scutellariae (RS) and Rhei Radix (RRR), which has been mainly used for treating migraine. AIM OF THE STUDY: This study aimed to uncover the anti-migraine active compounds from DJS and preliminary predicted the pharmacological mechanism by evaluating the spectrum-effect relationship between high-performance liquid chromatography (HPLC) fingerprints and anti-migraine effects of Duijinsan (DJS) extract combined with molecular docking. MATERIALS AND METHODS: HPLC and LC-MS were applied for chemical analyses of DJS extracts in different proportions. Inhibition of DJS extracts on trigeminal nerve cell releasing calcitonin gene related peptide (CGRP) experiment was performed. The active compounds were screened by spectrum-effect relationship analysis and confirmed by molecular docking and the activities of major predicted compounds were validated in vitro. RESULTS: Twenty-six common peaks were assigned and identified from the fingerprints of different proportions DJS extracts. In vitro experimental results showed that DJS extracts inhibited inflammation and release of CGRP from trigeminal nerve cells. Five predicted active compounds, Chrysin 6-C-arabinoside 8-C-glucoside, Chrysin 6-C-glucoside 8-C-arabinoside, baicalin, Chrysin-7-O-Beta-D-glucoronide and Oroxylin A 7-O-glucuronide were sorted out according to spectrum-effect relationship analysis and molecular docking comprehensively. In vitro validation experiments showed that all the predicted compounds inhibited the CGRP releasing and the activation of TRPV1 channel. Baicalin, chrysin-7-O-ß-D-glucuronide and Oroxylin A-7-glucoronide significantly inhibited the activation of TRPV1 channel. CONCLUSION: Chrysin 6-C-arabinoside 8-C-glucoside, Chrysin 6-C-glucoside 8-C-arabinoside, baicalin, Chrysin-7-O-Beta-D-glucoronide and Oroxylin A 7-O-glucuronide which can inhibit the CGRP releasing and the activation of TRPV1 channel were screened as the anti-migraine active compounds by spectrum-effect relationship analysis and molecular docking.
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Medicamentos de Ervas Chinesas/farmacologia , Transtornos de Enxaqueca/tratamento farmacológico , Rheum/química , Scutellaria baicalensis/química , Animais , Peptídeo Relacionado com Gene de Calcitonina/metabolismo , Células Cultivadas , Cromatografia Líquida de Alta Pressão , Cromatografia Líquida , Medicamentos de Ervas Chinesas/química , Células HEK293 , Humanos , Espectrometria de Massas , Simulação de Acoplamento Molecular , Ratos , Ratos Sprague-Dawley , Nervo Trigêmeo/citologia , Nervo Trigêmeo/efeitos dos fármacos , Nervo Trigêmeo/patologiaRESUMO
OBJECTIVE: To determine the therapeutic effectiveness and safety of polyethylene glycol 4000 (forlax) in the treatment of constipation in children over 8 years old. METHODS: This study was designed as a randomized, positive medicine (lactulose) controlled multicenter trial. A total of 216 children with constipation from 8-18 years old from 7 hospitals across China who were matched with a uniform entry criteria were enrolled in this study. The 216 patients were randomized to receive either oral forlax (20 g/d, n=105) or lactulose (15 mL/d, n=111) for 2 weeks. The therapeutic effects, including bowel movement frequency, stool consistency, clinical complete remission rate of constipation and abdominal symptoms, and the safety of forlax and lactulose were evaluated at 1 and 2 weeks of treatment. RESULTS: The median weekly frequency of bowel movement in the forlax group increased by 4 and 5 times respectively after 1 and 2 weeks of treatment, and increased by 3 and 4 times in the lactulose group (P < 0.05). The stool consistency of the two groups was both improved significantly after treatment. The Bristol score of stool consistency of the forlax and lactulose groups were 3.41+/-1.11 and 3.64+/-1.33 respectively (P < 0.05) after 1 week of treatment, and were 4.26+/-0.89 and 3.63+/-1.33 respectively (P < 0.05) after 2 weeks of treatment. The clinical complete remission rate of constipation in the forlax and lactulose groups was 70% and 40% respectively (P < 0.05) by week 1 of treatment, and that was 72% and 41% respectively (P < 0.05) by week 2 of treatment. Abdominal pain disappeared in 75% of patients in the forlax group but in only 57% in the lactulose group by week 2 of treatment (P < 0.05). No serious adverse events happened and no abnormalities were found in laboratory tests and physical examinations in the two groups after medication. CONCLUSIONS: Forlax is safe and effective in the treatment of constipation in children over 8 years old.
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Catárticos/uso terapêutico , Constipação Intestinal/terapia , Polietilenoglicóis/uso terapêutico , Adolescente , Catárticos/efeitos adversos , Criança , Feminino , Humanos , Lactulose/uso terapêutico , Masculino , Polietilenoglicóis/efeitos adversosRESUMO
This study aims to explore the correlation between microRNA-143 (miR-143) expression in peripheral blood mononuclear cells (PBMCs) and disease severity in patients with psoriasis vulgaris. From March 2014 to November 2015, 194 patients with psoriasis vulgaris (102 patients in progressive stage and 92 patients in stable stage) were selected as the case group and 175 healthy people as a control group were enrolled in this study. ELISA was used to detect the levels of IL-17 and VEGF in serum. The qRT-PCR assay was performed to detect the relative expression of miR-143 in PBMCs. Disease severity in psoriasis vulgaris patients was graded with Psoriasis Lesions Area and Severity Index (PASI). The value of miR-143 expression in PBMCs for the diagnosis of psoriasis vulgaris was evaluated using receiver operating characteristic (ROC) curve. The correlation between miR-143 expression in PBMCs and PASI scores was measured using Spearman rank correlation analysis. Compared with the control group, serum levels of IL-17 and VEGF were higher and miR-143 expression in PBMCs was lower in the case group. Furthermore, miR-143 expression in PBMCs was lower in patients in progressive stage than that in patients with stable stage. The relative expression of miR-143 in PBMCs was negatively correlated with PASI scores of patients with psoriasis vulgaris. ROC curve showed that miR-143 was a reliable and accurate biomarker of psoriasis vulgaris. Our findings suggest that miR-143 expression in PBMCs is negatively correlated the disease severity in psoriasis vulgaris.
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OBJECTIVE: To compare the clinical manifestations of gastroenteritis caused by norovirus and rotavirus in infants and young children in Beijing. METHODS: Stool specimens were collected from infants and young children with acute diarrhea who visited the Affiliated Children's Hospital to Capital Institute of Pediatrics from January 2002 to December 2006. Registration form was designed for clinical data collection for each patient from whom specimen was collected. Poly-acrylamide gel electrophoresis (PAGE) and enzyme immunoassay (EIA) were used to detect rotavirus and Human norovirus, respectively. RESULTS: Among 779 stool specimens tested for rotavirus, 263 were positive (33.8%), and norovirus positive specimens were 79 out of 318 (24.8%) specimens tested. Most of the clinical manifestations of gastroenteritis caused by these two viruses were quite similar with no significant difference, except for fever. The seasonal distribution of these two viruses were different with the peak of rotavirus infection was in cold weather between October and January, as indicated by the peak of the positive rates of the virus detection. The infection of norovirus seemed no obvious peak in the year. CONCLUSION: Rotavirus is the most important pathogen for acute diarrhea among infants and young children while. Norovirus is also an important pathogen for acute gastroenteritis in infants and young children. No significant difference was found out for clinical manifestations for the gastroenteritis caused by these two viruses.
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Diarreia/virologia , Gastroenterite/virologia , Norovirus/patogenicidade , Rotavirus/patogenicidade , Antígenos Virais , Infecções por Caliciviridae/virologia , Criança , Pré-Escolar , China , Fezes/virologia , Feminino , Genoma Viral , Humanos , Lactente , Masculino , Norovirus/isolamento & purificação , Rotavirus/isolamento & purificação , Infecções por Rotavirus/virologiaRESUMO
OBJECTIVE: To study the clinical manifestations for norovirus gastroenteritis in infants and young children. METHODS: Stool specimens were collected from infants and children with acute diarrhea who visited the affiliated Children's Hospital to Capital Institute of Pediatrics from January 2002 to December 2006. Enzyme-linked immunosorbent assay (ELISA) was used to detect human norovirus antigen in stool specimens and polyacrylamide gel electrophoresis (PAGE) was performed to detect rotavirus genome. RESULTS: Out of the 318 specimens under testing, 79 showed positive for norovirus antigen, with a positive rate of 24.8% (79/318). Among those positive specimens, 48(48/79, 60.8%) were detected in October to December, suggesting the seasonal preference of the virus. Most of the positive specimens (91.2%) were from those under 2 years of age. Rotavirus genome were detected from 16 out of 79 norovirus positive specimens (16/79, 20.3%), indicating those patients were co-infected by these two viruses. There was significant difference found in the severity of fever but not in the frequencies of diarrhea between rotavirus and norovirus co-infection group and noroviral infection group. Fourteen out of 79 norovirus positive patients were admitted to hospitals under the diagnosis other than gastroenteritis but started to develop symptoms of diarrhea between 1 to 11 days after hospitalization. CONCLUSION: Norovirus seemed one of the most important pathogens for acute diarrhea among infants and young children and could cause nosocomial infectious gastroenteritis.
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Infecções por Caliciviridae/diagnóstico , Infecções por Caliciviridae/virologia , Gastroenterite/diagnóstico , Gastroenterite/virologia , Norovirus/patogenicidade , Criança , Pré-Escolar , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Feminino , Genoma Viral/genética , Humanos , Lactente , Masculino , Norovirus/classificação , Norovirus/genéticaRESUMO
OBJECTIVE: In order to find out a more convenient, rapid and efficient way in detecting human Norovirus infections in specimens collected from hospitalized patients with acute non-bacterial diarrhea in Beijing. METHODS: Two kits for enzyme immunoassay (EIA) were used to detect human Noroviruses in stool specimens collected from 69 infants and young children as well as 15 adults who were all diagnosed as acute non-bacterial diarrhea in 4 different hospitals. Reverse transcription-polymerase chain reaction(RT-PCR) was performed to evaluate the data from two kits in this study. Data were statistically analyzed by SPSS 11.5 software. chi2 test was used to test categorical variables. RESULTS: Out of 84 stool specimens collected from infants and young children or adults with acute non-bacterial diarrhea, 17 (20.2%) were Norovirus positive determined by EIA kit A and 31 (36.9%) were Norovirus positive determined by EIA kit B. chi2 test used to test categorical variables showed significant differences (P < 0.01), suggesting that the EIA kit B was superior to the EIA kit A. Among these 84 stool specimens, 20 were tested by RT-PCR simultaneously. Out of those 20 specimens, 11 (55.0%) were Norovirus positive as determined by RT-PCR, which was higher than that from 2 EIA kits. RESULTS: from 10 (50.0%) samples detected by EIA kit A were consistent with those detected by RT-PCR. Through chi2 test, the categorical variables showed significant differences with P < 0.05, suggesting that RT-PCR was superior to the EIA kit A. Results from 14 (70.0%) samples detected by EIA kit B were consistent with those detected by RT-PCR while chi2 test showed that the differences were not significant (P > 0.05), among categorical variables suggesting that EIA kit B was as sensitive as RT-PCR in detecting Norovirus. The were hospital acquired diarrhea outbreaks in these three hospitals since at least 2 Norovirus positive specimens were detected in each of the hospitals. CONCLUSION: To detect human Noroviruses infection, EIA seemed to be more convenient and time saving than RT-PCR which had been used worldwide. The EIA kit B in this study was comparable to RT-PCR for detecting Norovirus in stool specimens. Norovirus was a pathogen causing hospital acquired diarrhea outbreaks in these three hospitals.
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Infecções por Caliciviridae/diagnóstico , Diarreia/virologia , Norovirus/genética , Adulto , Criança , Pré-Escolar , China , Infecção Hospitalar/diagnóstico , Fezes/virologia , Hospitais , Humanos , Técnicas Imunoenzimáticas , Lactente , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
OBJECTIVES: To develop techniques which can be used to detect genetic material of measles virus from clinical samples to make diagnosis and differentiate atypical measles from other exanthematous infections early in clinical course. METHODS: A nested reverse transcriptase-polymerase chain reaction (RT-PCR) was developed to amplify gene fragment with the size of 301 bps from N gene of measles virus in throat swabs and urine samples collected from infants and children who were suspected measles cases. Before the test was used for clinical samples, preliminary tests were performed to determine the sensitivity and specificity of the test. The sensitivity of the test was determined by plaque assay using measles virus strain Edmonton and the specificity of the test was determined by cross-reaction with rubella virus, respiratory syncytial virus, influenza A and B viruses, enterovirus, adenovirus, human cytomegalovirus (hCMV), EB virus, and herpes simplex virus I. Serum specific IgM antibody against measles virus was also tested by ELISA. RESULTS: Measles virus with the titer of 0.53 pfu could be detected by using the nested RT-PCR developed in this study. No amplification was found with the nested RT-PCR when rubella virus, respiratory syncytial virus, influenza A and B virus, enterovirus, adenovirus, hCMV, EB virus, and herpes simplex virus I were used as templates. Out of 116 throat swabs collected from suspected measles cases, 70 (60.3%) were measles RNA positive. For urine samples, 48 out of 74 (64.9%) were positive. Both throat swab and urine samples were collected simultaneously from 73 patients. Among those, 71 (97.3%) showed consistent results. Serum specimens were collected from 110 suspected patients. Among those, 65 (59.1%) and 61 (55.5%) were measles virus specific IgM antibody positive detected with ELISA kits from two different sources, respectively. Out of 110 sera samples, 106 (96.4%) showed consistent results. The consistency of the gene amplification and specific IgM antibody detection was 80.8% as shown by 84 out of 104 patients from whom throat swab and sera were collected at the same time. CONCLUSION: The data indicate that the nested RT-PCR developed in this study is sensitive and specific for detection of gene fragment of measles virus from clinical samples. The test is superior to the commonly used specific IgM antibody detection because of identifying gene material in early clinical stage, and even single clinical sample can be tested.