Postweaning multisystemic wasting syndrome (PMWS) is an important swine disease that is closely associated with porcine circovirus type 2 (PCV2). The capsid protein (Cap protein) is a major structural protein that has...Postweaning multisystemic wasting syndrome (PMWS) is an important swine disease that is closely associated with porcine circovirus type 2 (PCV2). The capsid protein (Cap protein) is a major structural protein that has at least three immunoreactive regions, and it can be a suitable candidate antigen for detecting the specific antibodies of a PCV2 infection. In the present study, an indirect enzyme-linked immunosorbent assay (TcELISA) based on a truncated soluble Cap protein produced in Escherichia coli (E.coli) was established and validated for the diagnostic PCV2 antibodies in swine. The TcELISA was validated by comparison with an indirect immunofluorescence assay (IIFA). The diagnostic sensitivity (DSN), specificity (DSP), and accuracy of the TcELISA were 88.6%, 90.7% and 89.4%, respectively. The agreement rate was 89.38% between results obtained with TcELISA and IIFA on 113 field sera. A cross-reactivity assay showed that the method was PCV2-specific by comparison with other sera of viral disease. Therefore ,the TcELISA will be helpful for the development of a reliable serology diagnostic test for large scale detection of PCV2 antibodies and for the evaluation of vaccine against PCV2 in swine.展开更多
Ontology is defined as an explicit specification of a conceptualization. In this paper, an extended ontology model was constructed using description logics, which is a 5-tuples including term set, individual set, term...Ontology is defined as an explicit specification of a conceptualization. In this paper, an extended ontology model was constructed using description logics, which is a 5-tuples including term set, individual set, term definition set, instantiation assertion set and term restriction set. Based on the extended model, the issue on ontology checking was studied with the conclusion that the four kinds of term checking, including term satisfiability checking, term subsumption checking, term equivalence checking and term disjointness checking, can be reduced to the satisfiability checking, and satisfiability checking can be transformed into instantiation consistence checking.展开更多
AIM: To explore epigenetic changes in the gene encod- ing X chromosome-linked inhibitor of apoptosis-associ- ated factor 1 (XAF1) during esophageal carcinogenesis. METHODS: Methylation status of XAF1 was detected ...AIM: To explore epigenetic changes in the gene encod- ing X chromosome-linked inhibitor of apoptosis-associ- ated factor 1 (XAF1) during esophageal carcinogenesis. METHODS: Methylation status of XAF1 was detected by methylation-specific polymerase chain reaction (MSP) in four esophageal cancer cell lines (KYSE30, KYSE70, BICl and partially methylated in TE3 cell lines), nine cases of normal mucosa, 72 cases of pri- mary esophageal cancer and matched adjacent tissue. XAF1 expression was examined by semi-quantitative reverse transcriptional polymerase chain reaction and Western blotting before and after treatment with 5-aza- deoxycytidine (5-aza-dc), a demethylating agent. To investigate the correlation of XAF1 expression and methylation status in primary esophageal cancer, immu- nohistochemistry for XAF1 expression was performed in 32 cases of esophageal cancer and matched adjacent tissue. The association of methylation status and clini-copathological data was analyzed by logistic regression. RESULTS: MSP results were as follows: loss of XAF1 expression was found in three of four esophageal cell lines with promoter region hypermethylation (com- pletely methylated in KYSE30, KYSE70 and BIC1 cell lines and partially in TE3 cells); all nine cases of normal esophageal mucosa were unmethylated; and 54/72 (75.00%) samples from patients with esophageal can- cer were methylated, and 25/72 (34.70%) matched adjacent tissues were methylated (75.00% vs 34,70%, z2 = 23.5840, P = 0.000). mRNA level of XAF1 mea- sured with semi-quantitative reverse transcription poly- merase chain reaction was detectable only in TE3 cells, and no expression was detected in KYSE30, KYSE70 or BIC1 cells. Protein expression was not observed in KYSE30 cells by Western blotting before treatment with 5-aza-dc. After treatment, mRNA level of XAF1 was detectable in KYSE30, KYSE70 and BIC1 cells. Protein expression was detected in KYSE30 after treatment with 5-aza-dc. Immunohistochemistry was performed on 32 cases of esophageal cancer and adjacent tissue, and demonstrated XAF1 in the nucleus and cytoplasm. XAF1 staining was found in 20/32 samples of adjacent normal tissue but was present in only 8/32 samples of esophageal cancer tissue (Z2= 9.143, P = 0.002). XAF1 expression was decreased in cancer samples compared with adjacent tissues. In 32 cases of esophageal can- cer, 24/32 samples were methylated, and 8/32 esopha- geal cancer tissues were unmethylated. XAF1 staining was found in 6/8 samples of unmethylated esophageal cancer and 2/24 samples of methylated esophageal cancer tissue. XAF1 staining was inversely correlated with XAF1 promoter region methylation (Fisher's exact test, P = 0.004). Regarding methylation status and clinicopathological data, no significant differences were found in sex, age, tumor size, tumor stage, or metas- tasis with respect to methylation of XAF1 for the 72 tis- sue samples from patients with esophageal cancer. CONCLUSION: XAF1 is frequently methylated in eso- phageal cancer, and XAF1 expression is regulated by promoter region hypermethylation.展开更多
Mycoplasma synoviae (MS) is an economically important pathogenic agent of chickens, causing air sacculitis and synovitis. The diagnosis and monitoring of M. synoviae infection is usually made using serological assay...Mycoplasma synoviae (MS) is an economically important pathogenic agent of chickens, causing air sacculitis and synovitis. The diagnosis and monitoring of M. synoviae infection is usually made using serological assays, while confirmative diagnosis is made by isolation and identifcation of the organism, because of the cross reaction between M. gallisepticum and M. synoviae. This study was conducted from 2011 to 2013 during which l l broiler breeder flocks were sampled. These farms were located in all regions of morocco, The sampling was conducted as follows: Farms were visited on day one old chicks "day of importation from Europe". 20 to 60 "chicks" were randomly sampled. At the age of 8, 16, 32 and 56 weeks, 60 blood samples and 60 tracheal swabs were collected at each sampling. The serological screening was performed using Rapid Slide Agglutination (RSA) according the OIE protocol and Indirect EL1SA (IDEXX) according the manufacturer's instructions. The molecular diagnosis was performed using a commercial kit of a duplex real time PCR (Life Biotechnology). The results revealed that one day old chicks were negative to MS by RSA and PCR, however they have a variable stock of maternal antibodies (Ig-Y) detected by iELISA. The seroprevalence found by RSA is variable and increase with the age (Sth week: 55%, 16th week: 91%, 32tb and 56th week: 100%), the same profile was traced by PCR (Sth week: 36%, 16th week: 64%, 32th week: 82%, 56th week: 100%), however, all farms were positive by iELISA, from the first day to 56th weeks. These results show that MS infections are very frequent and very widespread among poultry breeder flocks, and showed a perfect agreement between serological tests and Real time PCR starting from 32th week of age.展开更多
Objective:The aim of this study was to explore the relationship between follicular carcinoma of thyroid in different risk groups and lymph node micrometastases.Methods:The keratin-19 of negative neck lymph nodes in 83...Objective:The aim of this study was to explore the relationship between follicular carcinoma of thyroid in different risk groups and lymph node micrometastases.Methods:The keratin-19 of negative neck lymph nodes in 83 cases in routine pathological examination,was detected by SP immunohistochemistry using keratin-19 monoclonal antibody to confirm lymph node micrometastases.All of cases are divided into high risk group,middle risk group and low risk group according to factors related prognosis,the relationship between lymph node micrometastases and different risk groups and follow-up visit documents were analyzed.Results:Fifty-eight neck lymph nodes in 16 cases of 83 cases(19.3%) showed positive lymph node micrometastases,and incidence of lymph node micrometastases was 4/39 in low risk group,5/32 in middle risk group and 7/12 in high risk group,respectively.it showed remarkable difference during 3 groups(P < 0.001).Nine patients in 16 cases with positive lymph node micrometastases showed local recurrence and distant metastases,6 patients in 67 cases with negative lymph node micrometastases showed same result(P < 0.001).Conclusion:Lymph node micrometastases in follicular thyroid carcinoma closely correlated to factors related to prognosis.The detection of lymph node micrometastases can directly assistant postoperative treatment and prognosis evaluation to some extent for follicular thyroid carcinoma.展开更多
Coronavirus disease 2019(COVID-19),caused by SARS-CoV-2,has rapidly spread and caused a severe global pandemic.Because no specific drugs are available for COVID-19 and few vaccines are available for SARS-CoV-2,accurat...Coronavirus disease 2019(COVID-19),caused by SARS-CoV-2,has rapidly spread and caused a severe global pandemic.Because no specific drugs are available for COVID-19 and few vaccines are available for SARS-CoV-2,accurate and rapid diagnosis of COVID-19 has been the most crucial measure to control this pandemic.Here,we developed a portable bifunctional electrical detector based on graphene fieldeffect transistors for SARS-CoV-2 through either nucleic acid hybridization or antigen-antibody protein interaction,with ultra-low limits of detection of~0.1 and~1 fg mL^(−1) in phosphate buffer saline,respectively.We validated our method by assessment of RNA extracts from the oropharyngeal swabs of ten COVID-19 patients and eight healthy subjects,and the IgM/IgG antibodies from serum specimens of six COVID-19 patients and three healthy subjects.Here we show that the diagnostic results are in excellent agreement with the findings of polymerase chain reaction-based optical methods;they also exhibit rapid detection speed(~10 min for nucleic acid detection and~5 min for immunoassay).Therefore,our assay provides an efficient,accurate tool for high-throughput point-of-care testing.展开更多
基金supported by a project from National Key Technology R&D Program in the 11th Fiveyear Plan of China (2006BAD06A12)
文摘Postweaning multisystemic wasting syndrome (PMWS) is an important swine disease that is closely associated with porcine circovirus type 2 (PCV2). The capsid protein (Cap protein) is a major structural protein that has at least three immunoreactive regions, and it can be a suitable candidate antigen for detecting the specific antibodies of a PCV2 infection. In the present study, an indirect enzyme-linked immunosorbent assay (TcELISA) based on a truncated soluble Cap protein produced in Escherichia coli (E.coli) was established and validated for the diagnostic PCV2 antibodies in swine. The TcELISA was validated by comparison with an indirect immunofluorescence assay (IIFA). The diagnostic sensitivity (DSN), specificity (DSP), and accuracy of the TcELISA were 88.6%, 90.7% and 89.4%, respectively. The agreement rate was 89.38% between results obtained with TcELISA and IIFA on 113 field sera. A cross-reactivity assay showed that the method was PCV2-specific by comparison with other sera of viral disease. Therefore ,the TcELISA will be helpful for the development of a reliable serology diagnostic test for large scale detection of PCV2 antibodies and for the evaluation of vaccine against PCV2 in swine.
基金National Natural Science Foundation ofChina(No.70 2 710 3 8)
文摘Ontology is defined as an explicit specification of a conceptualization. In this paper, an extended ontology model was constructed using description logics, which is a 5-tuples including term set, individual set, term definition set, instantiation assertion set and term restriction set. Based on the extended model, the issue on ontology checking was studied with the conclusion that the four kinds of term checking, including term satisfiability checking, term subsumption checking, term equivalence checking and term disjointness checking, can be reduced to the satisfiability checking, and satisfiability checking can be transformed into instantiation consistence checking.
基金Supported by Grants from the National Basic Research Program(973 Program), No. 2012CB934002, 2010CB912802National Key Scientific Instrument Special Programme of China, No.2011YQ03013405National Science Foundation of China,No. 81121004, 81071953 and 81161120432
文摘AIM: To explore epigenetic changes in the gene encod- ing X chromosome-linked inhibitor of apoptosis-associ- ated factor 1 (XAF1) during esophageal carcinogenesis. METHODS: Methylation status of XAF1 was detected by methylation-specific polymerase chain reaction (MSP) in four esophageal cancer cell lines (KYSE30, KYSE70, BICl and partially methylated in TE3 cell lines), nine cases of normal mucosa, 72 cases of pri- mary esophageal cancer and matched adjacent tissue. XAF1 expression was examined by semi-quantitative reverse transcriptional polymerase chain reaction and Western blotting before and after treatment with 5-aza- deoxycytidine (5-aza-dc), a demethylating agent. To investigate the correlation of XAF1 expression and methylation status in primary esophageal cancer, immu- nohistochemistry for XAF1 expression was performed in 32 cases of esophageal cancer and matched adjacent tissue. The association of methylation status and clini-copathological data was analyzed by logistic regression. RESULTS: MSP results were as follows: loss of XAF1 expression was found in three of four esophageal cell lines with promoter region hypermethylation (com- pletely methylated in KYSE30, KYSE70 and BIC1 cell lines and partially in TE3 cells); all nine cases of normal esophageal mucosa were unmethylated; and 54/72 (75.00%) samples from patients with esophageal can- cer were methylated, and 25/72 (34.70%) matched adjacent tissues were methylated (75.00% vs 34,70%, z2 = 23.5840, P = 0.000). mRNA level of XAF1 mea- sured with semi-quantitative reverse transcription poly- merase chain reaction was detectable only in TE3 cells, and no expression was detected in KYSE30, KYSE70 or BIC1 cells. Protein expression was not observed in KYSE30 cells by Western blotting before treatment with 5-aza-dc. After treatment, mRNA level of XAF1 was detectable in KYSE30, KYSE70 and BIC1 cells. Protein expression was detected in KYSE30 after treatment with 5-aza-dc. Immunohistochemistry was performed on 32 cases of esophageal cancer and adjacent tissue, and demonstrated XAF1 in the nucleus and cytoplasm. XAF1 staining was found in 20/32 samples of adjacent normal tissue but was present in only 8/32 samples of esophageal cancer tissue (Z2= 9.143, P = 0.002). XAF1 expression was decreased in cancer samples compared with adjacent tissues. In 32 cases of esophageal can- cer, 24/32 samples were methylated, and 8/32 esopha- geal cancer tissues were unmethylated. XAF1 staining was found in 6/8 samples of unmethylated esophageal cancer and 2/24 samples of methylated esophageal cancer tissue. XAF1 staining was inversely correlated with XAF1 promoter region methylation (Fisher's exact test, P = 0.004). Regarding methylation status and clinicopathological data, no significant differences were found in sex, age, tumor size, tumor stage, or metas- tasis with respect to methylation of XAF1 for the 72 tis- sue samples from patients with esophageal cancer. CONCLUSION: XAF1 is frequently methylated in eso- phageal cancer, and XAF1 expression is regulated by promoter region hypermethylation.
文摘Mycoplasma synoviae (MS) is an economically important pathogenic agent of chickens, causing air sacculitis and synovitis. The diagnosis and monitoring of M. synoviae infection is usually made using serological assays, while confirmative diagnosis is made by isolation and identifcation of the organism, because of the cross reaction between M. gallisepticum and M. synoviae. This study was conducted from 2011 to 2013 during which l l broiler breeder flocks were sampled. These farms were located in all regions of morocco, The sampling was conducted as follows: Farms were visited on day one old chicks "day of importation from Europe". 20 to 60 "chicks" were randomly sampled. At the age of 8, 16, 32 and 56 weeks, 60 blood samples and 60 tracheal swabs were collected at each sampling. The serological screening was performed using Rapid Slide Agglutination (RSA) according the OIE protocol and Indirect EL1SA (IDEXX) according the manufacturer's instructions. The molecular diagnosis was performed using a commercial kit of a duplex real time PCR (Life Biotechnology). The results revealed that one day old chicks were negative to MS by RSA and PCR, however they have a variable stock of maternal antibodies (Ig-Y) detected by iELISA. The seroprevalence found by RSA is variable and increase with the age (Sth week: 55%, 16th week: 91%, 32tb and 56th week: 100%), the same profile was traced by PCR (Sth week: 36%, 16th week: 64%, 32th week: 82%, 56th week: 100%), however, all farms were positive by iELISA, from the first day to 56th weeks. These results show that MS infections are very frequent and very widespread among poultry breeder flocks, and showed a perfect agreement between serological tests and Real time PCR starting from 32th week of age.
文摘Objective:The aim of this study was to explore the relationship between follicular carcinoma of thyroid in different risk groups and lymph node micrometastases.Methods:The keratin-19 of negative neck lymph nodes in 83 cases in routine pathological examination,was detected by SP immunohistochemistry using keratin-19 monoclonal antibody to confirm lymph node micrometastases.All of cases are divided into high risk group,middle risk group and low risk group according to factors related prognosis,the relationship between lymph node micrometastases and different risk groups and follow-up visit documents were analyzed.Results:Fifty-eight neck lymph nodes in 16 cases of 83 cases(19.3%) showed positive lymph node micrometastases,and incidence of lymph node micrometastases was 4/39 in low risk group,5/32 in middle risk group and 7/12 in high risk group,respectively.it showed remarkable difference during 3 groups(P < 0.001).Nine patients in 16 cases with positive lymph node micrometastases showed local recurrence and distant metastases,6 patients in 67 cases with negative lymph node micrometastases showed same result(P < 0.001).Conclusion:Lymph node micrometastases in follicular thyroid carcinoma closely correlated to factors related to prognosis.The detection of lymph node micrometastases can directly assistant postoperative treatment and prognosis evaluation to some extent for follicular thyroid carcinoma.
基金the National Key R&D Program of China(2017YFA0204901)the National Natural Science Foundation of China(21727806,21772003 and 21933001)+1 种基金the Tencent Foundation through the XPLORER PRIZE,Guangdong Major Project of Basic and Applied Basic Research(2019B030302007)Beijing National Laboratory for Molecular Sciences(BNLMS201901)。
文摘Coronavirus disease 2019(COVID-19),caused by SARS-CoV-2,has rapidly spread and caused a severe global pandemic.Because no specific drugs are available for COVID-19 and few vaccines are available for SARS-CoV-2,accurate and rapid diagnosis of COVID-19 has been the most crucial measure to control this pandemic.Here,we developed a portable bifunctional electrical detector based on graphene fieldeffect transistors for SARS-CoV-2 through either nucleic acid hybridization or antigen-antibody protein interaction,with ultra-low limits of detection of~0.1 and~1 fg mL^(−1) in phosphate buffer saline,respectively.We validated our method by assessment of RNA extracts from the oropharyngeal swabs of ten COVID-19 patients and eight healthy subjects,and the IgM/IgG antibodies from serum specimens of six COVID-19 patients and three healthy subjects.Here we show that the diagnostic results are in excellent agreement with the findings of polymerase chain reaction-based optical methods;they also exhibit rapid detection speed(~10 min for nucleic acid detection and~5 min for immunoassay).Therefore,our assay provides an efficient,accurate tool for high-throughput point-of-care testing.