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Qing Duan,
Hong Zhu,
Yi Yang,
Weihua Li,
Yusen Zhou,
Jun He,
Kun He,
Haojie Zhang,
Tao Zhou,
Lihua Song,
Yonghua Gan,
Hua Tan,
Baofeng Jin,
Huiyan Li,
Tingting Zuo,
Dehui Chen,
Xuemin Zhang
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ABSTRACT: Beijing has been severely affected by SARS, and SARS-associated coronavirus has been confirmed as its cause. However, clinical
and experimental evidence implicates the possibility of co-infection. In this report, reovirus was isolated from throat swabs
of SARS patients, including the first case in Beijing and her mother. Identification with the electron microscopy revealed
the characteristic features of reovirus. 24 of 38 samples from other SARS cases were found to have serologic responses to
the reovirus. Primers designed for reovirus have amplified several fragments of DNA, one of which was sequenced (S2 gene fragment),
which indicates it as a unique reovirus (orthoreovirus). Preliminary animal experiment showed that inoculation of the reovirus
in mice caused death with atypical pneumonia. Nevertheless, the association of reovirus with SARS outbreak requires to be
further investigated.
Keywordsreovirus-coronavirus-SARS-electron microscopy
Chinese Science Bulletin 04/2012; 48(13):1293-1296. · 1.32 Impact Factor
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ABSTRACT: To define the sequence type (ST) isolates of Bacillus anthracis by multilocus sequence typing (MLST).
Fragments of seven housekeeping genes (glpF, gmk, ilvD, pta, pur, pycA, and tpi) were amplified by PCR using the standard primers as described on the website for MLST of Bacillus and the sequences were compared with existing allele sequences on the MLST website.
Two novel allele combinations of the seven loci were found in two isolates 17003-14 and 17003-32.
Two novel ST isolates of B. anthracis were identified by this study and confirmed by the MLST website, and the pubMLST ids were id-1053 and id-1054.
ACTA MICROBIOLOGICA SINICA 01/2012; 52(1):120-3.
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ABSTRACT: The main aim of this study was to develop and optimize a questionnaire-based zygosity determination method in Chinese adolescent twins. Participants were 471 pairs of same-sex twins (345 monozygotic, 126 dizygotic) with a mean age of 14.56 years (SD=2.62). A second sample was recruited for cross-validation, including 382 pairs of same-sex twins (261 monozygotic, 121 dizygotic) with a mean age of 12.53 years (SD=2.22). The questionnaire consisted of 12 questions dealing with co-twin similarity or frequency of confusion. Two means were put forward to improve the predictive accuracy of the questionnaire--adding parent-reports to the analysis, and using a 2-point rather than 3-point response format. DNA genotyping was performed on nine short tandem repeat loci, with an estimated zygosity classification accuracy very close to 100%. The validity of all questionnaires was assessed by being compared to the results of DNA analysis. Results of stepwise logistic regression analysis showed that the predictive accuracy of the 3-point self-reported questionnaire is 83.8%. Using parent-reports and 2-point scale led to 3.9% and 4.6% increase in predictive accuracy, respectively. When using the parent-reports and children's self-reports jointly, the predictive accuracy was enhanced to 90.6%. For the cross-validation, the equations and cut-offs derived from the first sample led to an acceptable accuracy (91.3%) in the second sample. In conclusion, the method we developed can be used in future studies among Chinese adolescent twins. Multiple-rater and 2-point response format were suggested for all twin studies for enhancing the predictive accuracy of questionnaires.
Twin Research and Human Genetics 04/2010; 13(2):194-200. · 1.70 Impact Factor
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ABSTRACT: Adolescents' romantic relationships have been associated with higher levels of depression, although their links with externalizing behavioral problems remain unclear. The present study examined the impact of adolescent romantic relationships on depression and externalizing behaviors in a large sample of 10,509 Chinese secondary school students (ages 12-19, 54.5% female). The results showed that romantic involvement in adolescence, especially in early adolescence, was associated with more depressive symptoms and behavior problems. Breakups in romantic relationships were an important factor in producing the negative emotional and behavioral consequences. Romantically involved girls experienced higher levels of depressive symptoms, while romantically involved boys had higher levels of externalizing behaviors, compared to their non-dating peers. The results also indicated that the adverse impact was stronger for those involved in romantic relationships at younger ages.
Journal of Youth and Adolescence 11/2009; 38(10):1282-93. · 2.72 Impact Factor
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Clinical Infectious Diseases 09/2009; 49(3):480. · 9.15 Impact Factor
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ABSTRACT: We previously described isolation of a potentially new mammalian reovirus, designated BYD1, which can cause clinical symptoms similar to that of severe acute respiratory syndrome (SARS) in guinea pigs and macaques, from throat swabs of one SARS patient of Beijing, in 2003. For this study, we determined the genome sequences of BYD1 and the S1 gene sequences of other five mammalian reovirus isolates (BLD, JP, and BYL were isolated from different SARS patients during the outbreak, 302I and 302II were isolated from fecal specimens of two children of Beijing in 1982) to allow molecular comparison with other previously reported mammalian reoviruses (MRVs). Comparative analyses of the BYD1 genome with those of three prototype mammalian reovirus strains demonstrated that BYD1 is a novel reassortant virus, with its S1 gene segment coming from a previously unidentified serotype 2 isolate and other nine segments coming from ancestors of homologous T1L and T3D segments, which supports the hypothesis that mammalian reovirus gene segments reassort in nature. Further analyses of the S1 segments of the six isolates showed that all the isolates are novel serotype 2 MRVs based on their S1 gene sequences, which are markedly different from those of all previously reported, and the S1 genes of the four new isolates share more than 99% identity with each other, proving that they diverged from a common ancestor most recently, and the S1 genes of the four new isolates share about 65% identity with those of the two strains isolated in 1982.
Virus Genes 10/2008; 37(3):392-9. · 1.85 Impact Factor
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Hong-Bo Jing,
Jing Yuan,
Jie Wang,
Yuan Yuan,
Li Zhu,
Xian-Kai Liu,
Yu-Ling Zheng,
Kai-Hua Wei,
Xue-Min Zhang,
Hong-Ran Geng, Qing Duan,
Shu-Zhang Feng,
Rui-Fu Yang,
Wu-Chun Cao,
Heng-Liang Wang,
Yong-Qiang Jiang
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ABSTRACT: Outbreaks in humans, caused by Streptococcus suis serotype 2 (SS2), were reported in 1998 and 2005 in China. However, the mechanism of SS2-associated infection remains unclear. For the first time, a 2-D gel approach combined with MS was used to establish a comprehensive 2-D reference map for aiding our understanding of the pathogenicity of SS2. The identification of 694 out of 834 processed spots revealed 373 proteins. Most of the identified proteins were located in the cytoplasm and were involved in energy metabolism, protein synthesis, and cellular processes. Proteins that were abundant in the 2-DE gels could be linked mainly to housekeeping functions in carbohydrate metabolism, protein quality control and translation. 2-DE of secretory proteins was performed using IPG strips of pH 4-7. Among the 102 protein spots processed, 87 spots representing 77 proteins were successfully identified. Some virulence-associated proteins of SS2 were found, including arginine deiminase, ornithine carbamoyl-transferase, carbamate kinase, muramidase-released protein precursor, extracellular factor, and suilysin. Enolase and endopeptidase have been proposed as putative virulence-associated factors in this study. The 2-D reference map might provide a powerful tool for analyzing the virulence factor and the regulatory network involved in the pathogenicity of this microorganism.
PROTEOMICS 02/2008; 8(2):333-49. · 4.51 Impact Factor
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ABSTRACT: To study a newly isolated domestic mammalian reovirus, BYD1, its ability to induce apoptosis analyze the three-dimensional structure of its major membrane penetration protein to predict its function in inducing apoptosis.
HeLa cells infected with BYD1 reovirus were metered with flow cytometer (FCM) to quantify the ratio of apoptotic cells. The data were analyzed with Student's t-test to judge the ability of BYD1 strain to induce apoptosis. The primary sequence ranged from 582 to 675 per microliter protein of BYD1, T1L, T2J and T3D were aligned and compared. The three-dimensional comparative protein structure model of microliter protein was generated by homology-modeling pipeline SWISS MODEL was applied to annotate its secondary and tertiary structure.
BYD1 strain was verified with the ability to induce the apoptosis of HeLa cells. The 643-675 segment composing an alpha-helix showed major difference compared with prototype T2J.
The newly isolated reovirus BYD1 is an apoptosis inducing strain. The alpha-helix (residues 643 to 675) of microliter protein of BYD1 may play a key role to induce the proapoptotic activity of infected cells.
Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang bingduxue zazhi = Chinese journal of experimental and clinical virology 10/2007; 21(3):223-5.
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ABSTRACT: The objective of this study was to investigate the pathogenicity and associated lesions of a new reovirus (ReoV) isolated from patients with Severe Acute Respiratory Syndrome (SARS) in China. Twenty-five four-week-old BALB/c female mice inoculated intranasally with either ReoV (strain BYD1) alone, or ReoV combined with SARS-CoV (strain BJF) displayed ejecting fur and loss of body weight compared with control animals. ReoV and SARS-CoV were isolated from most postmortem tissues. The histopathological features of ReoV infected animals consisted of diffuse alveolar damage, with scattered hemorrhage, hyaline membrane formation and interstitial pneumonia. A typical type II pneumocyte hyperplasia and fibrogranulomatous tissue formation in the alveolar septae were observed both in the animals inoculated simultaneously with these two viruses and in the animals inoculated firstly with SARS-CoV, followed by ReoV. The animals inoculated firstly with ReoV, followed with SARS-CoV displayed scattered hemorrhage in the alveolar septa. Furthermore, other lesions in above two combination groups included depletion of lymphocytes in the germinal center of lymph nodes in the lung hilus and the spleen, hemorrhagic necrosis in white pulp of spleen, hydroid degeneration, and fatty degeneration in the liver and kidney. Mice induced with SARS-CoV alone did not display clinical signs, characteristically hyaline membrane formation, hemorrhage and early pulmonary fibrosis in lung tissue. This study demonstrated that the newly isolated ReoV might be a virulent pathogen for BALB/c mice. Mice infected firstly with SARS-CoV, followed with ReoV developed a typical diffuse alveolar lesion.
Experimental Animals 11/2006; 55(5):439-47. · 0.92 Impact Factor
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ABSTRACT: Streptococcus suis serotype 2 (SS2) is a major pathogen frequently associated with infections in pigs. There are presently 35 serotypes of S.suis (serotype 1 to 34 and serotype 1/2) recognized on the basis of capsular antigens. Few people were reported to infect with SS2 in the past years. However, an accidental case happened in Sichuan province of China in 2005. Some people got ill and died, and all of them were closely contacted with sick pigs. Based on clinical features and epidemiologic data, this case could be caused by SS2 infection. Liver, spleen, kidney, lung and serum samples were collected and used for pathogen isolation and identification in laboratory, three strain bacteria were isolated. The three strains of SS2 showed typical morphology of SS2 on blood agar and under microscope with Gram stain. They were also agglutinated with standard serum of SS2. Biochemical characteristics of the three bacteria were tested using API 20 strep and analyzed by API software (version 3.3), results showed they were SS2. Four pairs of primer were designed, which were exactly matched the extracellular factor gene, muraminidase released protein gene, capsular polysaccharides gene and 16S rRNA gene respectively. These primers were used on polymerase chain reaction (PCR), and the PCR products were 626bp, 885bp, 487bp and 297bp on agarose gel, respectively. Drug sensitivity test were also done and results showed that they were sensitive to cefazolin, clindamycin, erythromycin, levofloxacin, nitrofurantoin, penicillin-G, and vancomycin and resistive to tetracycline. Balb/c mice infected with the isolated SS2 strain showed swelling in stomach and intestine, cyanochroia at mouth and suggillation under skin, which were similar to the clinical features of patients. Streptococcus suis serotype 2 were also found on lung sheeting sample under microscope with Gram stain. Rabbits infected with the isolated SS2 showed the similar clinical features with mice.
ACTA MICROBIOLOGICA SINICA 09/2006; 46(4):635-8.
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Fang Zhang,
Wei Liu,
May C Chu,
Jun He, Qing Duan,
Xiao-Ming Wu,
Pan-He Zhang,
Qiu-Min Zhao,
Hong Yang,
Zhong-Tao Xin,
Wu-Chun Cao
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ABSTRACT: A total of 420 rodents in China were examined for Francisella tularensis by polymerase chain reaction. The infection rates were 4.76% in total, and 11.65%, 10.00%, 6.56%, 1.77%, and 0% in Jilin, Xinjiang, Heilongjiang, Inner Mongolia, and Zhejiang, respectively. Sequence analysis showed that all the detected agents belonged to F. tularensis subsp. holarctica.
Emerging infectious diseases 07/2006; 12(6):994-6. · 6.17 Impact Factor
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ABSTRACT: During June to July 2005, a few farmers in Chengde county of Hebei province were got ill after eating beef of sick cattle. The cattle could be infected with Bacillus anthracis. One beef sample and one soil sample contaminated with cattle blood were collected and used for pathogen isolation and identification in laboratory. Two bacteria strains were isolated from beef and soil sample, respectively, and showed typical morphology of Bacillus anthracis on blood agar and under microscope with Gram stain. The two bacteria strains were also positive to standard positive serum of Bacillus anthracis by slide agglutination test. Biochemical characteristics of the two bacteria were tested using API CHB/E strip and analyzed by API software (version 3.3), result showed that the two isolated bacteria were Bacillus anthracis. Polymerase chain reaction (PCR) was used to further characterize the two isolated bacteria strains. Three pairs of primer were designed and used for PCR, and these primers exactly matched the protective antigen gene, edema factor gene and capsule gene, respectively. By analyzed on agarose gel, PCR products were 423bp, 494bp and 397bp, respectively, and this result showed that the two isolated bacteria contained two plasmids, pX01 and pX02, which encoded anthrax toxin and capsule, respectively. Anthrax toxin and capsule were very important virulent factors for Bacillus anthracis. PCR products were purified and then cloned to T vector, positive clone was chose and sequenced. By BLAST with GenBank, sequence of the three genes of the two bacteria strains had a similarity of 99% with Bacillus anthracis A2012 strain, Ames Ancestor strain and A16R strain. Based on results of colonial morphology, serum test and biochemistry characterization, the two bacteria strains are Bacillus anthracis. They can encode anthrax toxin and capsule, and are virulent to animal and human.
ACTA MICROBIOLOGICA SINICA 07/2006; 46(3):460-2.
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ABSTRACT: To develop a method of immunochromatography assay (ICA) with sensitive, specific, rapid, simple and suitable for the detection of Yersinia pestis antigen at the local laboratories.
Colloidal gold labeled with the anti-F1 antibody of Yersinia pestis, was connnected with the anti-F1 antibody of Yersinia pestis to pyroxylin membrane and assembled them to the dipstick of ICA.
Results showed that the rates of sensitivity for F1 antigen and Yersinia pestis were 1 ng/ml and 1.56 x 10(5) CFU/ml respectively. However, Yersinia pseudotuberculosis et al could not be detected by dipstick of ICA.
The method of ICA appeared to be consistent to those of r-IHA with better specificity and sensitivity but was simple and rapid for the detection of Yersinia pestis and F1 antigen.
Zhonghua liu xing bing xue za zhi = Zhonghua liuxingbingxue zazhi 04/2006; 27(3):253-5.
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ABSTRACT: Experimental studies were performed to determine the role of a newly isolated reovirus (ReoV) from a severe acute respiratory syndrome (SARS) patient in the etiology of this newly described serious respiratory syndrome. Four cynomologus macaques were inoculated with this reovirus (BYD1) in an attempt to replicate the infection and pathology observed in SARS. The body temperature of the infected monkeys was monitored three times a day, and blood and fecal samples were periodically collected for specific immunology determinations. On days 7 and 33 after inoculation, necropsies for pathological accessment and pathogen isolation were performed. The four infected macaques developed a fever on days 3 and 4 after inoculation, and maintainted a febrile state for 4-6 days. The highest temperature in the animals recorded was 40.4 degrees C. After a recovery phase, the macaques developed a second febrile condition. Antibody titers against the reovirus injected by the intravenous route occurred in higher number than those in the nasal cavity. Four macaque monkeys demonstrated diffuse alveolar damage, characterized by hemorrhagic pneumonia, serosanguineous exudates, formation of hyaline membranes, and type II pneumocyte hyperplasia, which were similar to those that have been noted in SARS patients. Lymphocytes decreased in the cortex of the lymph node and in the white pulp of the spleen. ReoV was detected in pneumonic tissue by virus isolation and RT-PCR. The macaques infected with the newly isolated reovirus developed a fever, diffuse alveolar damage and pulmonary interstitial inflammation similar to that noted in SARS patients. This evidence demonstrates that ReoV might have a primary role in the etiology of SARS.
DNA and Cell Biology 09/2005; 24(8):491-5. · 2.07 Impact Factor
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ABSTRACT: Guinea pigs were inoculated with a reovirus (ReoV) and coronavirus (SARS-CoV) isolated from SARS patients to determine their potential role in the etiology of SARS. Animals infected with ReoV died between day 22 and day 30 postinoculation (PI) while 70% of the animals inoculated with ReoV and SARS-CoV died between day 4 to day 7 PI. The titer of neutralizing antibodies against ReoV and SARS-CoV ranged from 80 to 160 when the animals were inoculated with the two viruses, respectively, while the titer of the antibodies was just below 10 in coinfections. The animal inoculated with ReoV developed diffuse alveolar damage similar to the exudative and leakage inflammation found in SARS patients, and was characterized by diffuse hemorrhage, fibroid exudation, hyaline membrane formation, and type II pneumocytes hyperplasia in alveolar interstitia. The pulmonary epithelial necrosis, excoriation, and early fibrosis of pulmonary tissue were only observed in ReoV-SARS-CoV groups and in SARS-CoV/ReoV groups. Other typical pathological changes included hemorrhagic necrosis in lymph nodes and spleen and hydropic degeneration in the liver. On the contrary, guinea pigs infected with SARS-CoV only developed interstitial pneumonitis. Our experiment demonstrate that ReoV might be one of the primary causes of SARS, since simultaneous coinfection can duplicate the typical pathological changes similar to that of SARS patients. This guinea pig model may provide a useful animal model for SARS.
DNA and Cell Biology 09/2005; 24(8):485-90. · 2.07 Impact Factor
