Whole-genome analysis of porcine epidemic diarrhea virus (PEDV) from eastern China.

The complete genome sequence of a porcine epidemic diarrhea virus variant, strain SHQP/YM/2013, from China was determined and compared with those of other porcine epidemic diarrhea viruses. The full-length genome was 28,038 nucleotides (nt) in length without the poly (A) tail, and it was similar to that of other reported PEDV strains, with the characteristic gene order 5'-replicase (1a/1b) -S-ORF3-E-M-N-3'. Nucleotide sequence analysis based on individual virus genes indicated a close relationship between the S gene of SHQP/YM/2013 and those of the four Korean field strains from 2008-2009. Its ORF3 gene, however, fell into three groups. Recent prevalent Chinese PEDV field isolates were divided between group 1 and group 3, which suggests that the recent prevalent Chinese PEDV field isolates represent a new genotype that differs from the genotype that includes the vaccine strains. Based on phylogenetic analysis of the M gene, ORF3 gene and S gene, our study demonstrated that prevalent PEDV isolates in China may have originated from Korean strains. This report describes the complete genome sequence of SHQP/YM/2013, and the data will promote a better understanding of the molecular epidemiology and genetic diversity of PEDV field isolates in eastern China.

Epidemiological situation and genetic analysis of H7N9 influenza viruses in Shanghai in 2013.

The first reported human case of H7N9 influenza virus infection in Shanghai prompted a survey of local avian strains of influenza virus, involving the analysis of a large number of samples taken from poultry, wild birds, horses, pigs, dogs and mice. Seven instances of H7N9 virus infection were identified by real-time RT-PCR (1.47 % of samples), all in chickens sold in live-poultry markets. H7N9 antibody was not detected in serum samples collected from local poultry farms since 2006. The two H7N9 virus strains in the live-poultry markets and one H9N2 virus strain in the same market were genetically characterized. Resequencing of two of the seven isolates confirmed that they closely resembled H7N9 virus strains characterized elsewhere. Various strains co-exist in the same market, presenting a continuing risk of strain re-assortment. The closure of live-poultry markets has been an effective short-term means of minimizing human exposure to H7N9 virus.

Epidemiological survey of porcine epidemic diarrhea virus in swine farms in Shanghai, China.

An epidemiological survey of porcine diarrheal disease prevalence between September 2011 and January 2012 revealed that porcine epidemic diarrhea virus (PEDV) contributed to outbreaks of diarrhea in pig farms in Shanghai, China. The distribution profile of 10 PEDV strains revealed three distinct genotypes coexisting in the same pig farm. Two of the ten field strains that were isolated exhibited a distinct evolution from the others. In addition to PEDV, other enteric pathogens, including porcine kobuvirus, porcine teschovirus and Lawsonia intracellularis, were identified.

Epidemiological survey and genetic evolution of H9 subtype influenza viruses in Shanghai, China, from 2006 to 2010.

The H9N2 influenza virus is endemic in poultry. We report its occurrence in live-poultry markets, fair-trade markets and poultry farms in the Shanghai region between September 2006 and December 2010. An analysis of partial sequences of the HA, NA, PB1, PB2 and NP genes of eleven distinct H9N2 isolates revealed that all carried an RSSR motif at the cleavage site of HA, diagnostic of low pathogenicity in chickens. A phylogenetic analysis indicated that these isolates are derived from the lineage represented by Duck/HK/Y280/97, but they have evolved a range of reassortments. Their PB1 and NP sequences resembled those of H5N1 strains, indicating a hybrid origin involving both H9 and H5 strains. The HA and NA sequences present in all eleven isolates resembled those of the Duck/HK/Y280/97-like lineage. Infection by H9N2 is commonplace in Shanghai live-poultry markets, allowing the viruses to have evolved rapidly.

Genetic evolution of H9 subtype influenza viruses from live poultry markets in Shanghai, China.

H9N2 influenza viruses have become established and maintain long-term endemicity in poultry. The complete genomes of seven avian H9N2 influenza viruses were characterized. These seven influenza virus isolates were obtained from live poultry markets in Shanghai, China, in 2002 and from 2006 to 2008. Genetic analysis revealed that all seven isolates had an RSSR motif at the cleavage site of hemagglutinin (HA), indicating low pathogenicity in chickens. Phylogenetic analyses indicated that the seven avian H9N2 viruses belonged to the lineage represented by Duck/Hong Kong/Y280/97 (H9N2), a virus belonging to the Chicken/Beijing/1/94-like (H9N2) lineage, and that they are all quadruple reassortants consisting of genes from different lineages. The six internal genes of the isolates possessed H5N1-like sequences, indicating that they were reassortants of H9 and H5 viruses. All of the viruses had nonstructural (as well as HA and neuraminidase) genes derived from the Duck/Hong Kong/Y280/97-like virus lineage but also had other genes of mixed avian virus origin, including genes similar to those of H5N1 viruses (Gs/GD-like). The infected chickens showed no signs of disease. These results show the genetic and biological diversity of H9N2 viruses in Shanghai and support their potential role as pandemic influenza agents.

[Effect of Coptis Chinensis compound on the gene expression in transplanted tumor tissue in nasopharyngeal carcinoma cell line of CNE1 by cDNA microarray].

OBJECTIVE: To investigate the effect of Coptis Chinensis compound on the gene expression of transplanted tumor in nasopharyngeal carcinoma cell line of CNE1. METHODS: The cells were injected into Balb/C nude mice, and after the transplanted tumor was established, the nude mice with tumors were treated with Coptis chinensis compound. The nude mice were then killed, tumor tissues were isolated and the total RNA was extracted. Hybridization with a cDNA microarray was conducted using a probe of fluorescent-labeled cDNA mixture that was reversely transcribed from the RNA. The gene expression level was analyzed after scanning the fluorescent intensity by a laser scanner, and further confirmed by RT-PCR. RESULTS: After using Coptis Chinensis compound for 30 days, the size of transplanted tumors were remarkably reduced, and the inhibition rate of tumor growth was 29.5%. In the meantime, the expression of 147 genes in the tissue of implanted tumor treated with the compound altered: the expression of 102 genes was down-regulated, and that of 45 genes was up-regulated. Furthermore, the differential expression of 3 genes was verified by RT-PCR, MAD3 and H731 genes were up-regulated, and that of CHK1 was down-regulated. CONCLUSION: Coptis Chinensis compound can affect the gene expression in the tissue of implanted tumor, which implies that the inhibition of Coptis Chinensis compound on the growth of the implanted tumor with CNE1 cells in nude mice might be associated with the control of gene expression.

[The fluctuation of anterior teeth colouration during the period of growth puberty in children and adolescents].

OBJECTIVE: This study was designed to examine the fluctuation tendency of the anterior teeth coloration during the period of growth puberty. METHODS: One hundred children with average age of 13.3 years, and 100 adolescents with average age of 23.4 years were divided into two groups. The upper left incisors were selected for color matching using computerized spectrophotometery system (Shade-eye, Shofu). The measurements depicting color properties, including hue, value and chroma were quantitatively analyzed. Statistical evaluation was conducted for the comparison of the color measurements between the two groups. RESULTS: The following trends were indicated during the period of growth puberty: (1)The hue of the coloration changed from redness into mixture of redness and grayness. (2)The magnitude of value of the coloration was in decline. (3)The intensity of chroma increased significantly. CONCLUSIONS: The teeth coloration fluctuates within the period spanning the initial and final stages of growth puberty, characterized by a decrease in hue and value and an increase in chroma.

[In situ expression of connexins in various carcinomas].

BACKGROUND & OBJECTIVE: Malignant cells are often associated with aberrant expression or localization of connexins. Expression of Cx26, Cx32, Cx43, and Cx45 genes were studied to gain their expression profile in tissue microarrayers consisting of various carcinomas and to elucidate their function in carcinogenesis. METHODS: Basing on the principle of constructing tissue microarrays, the authors take use of self-made tissue arrayer to construct tissue microarrays for the different kinds of cancer specimens. Meanwhile, expression of connexin(Cx) genes, such as Cx32, Cx26, Cx43, and Cx45 in the tissue microarrays were detected in 292 cases of tumor tissues and 89 paratumor tissues by immunohistochemistry. RESULTS: (1) Three kinds of tissue microarrays consisting samples from 20 different carcinomas were constructed successfully, which contained 360, 200, or 100 spots in each receptive paraffin block with tissue cylinder of 0.75mm or 0.6mm diameter respectively.(2) The expression profiles of Cx32, Cx26, Cx43, and Cx45 in 20 kinds of carcinomas and their related adjacent-cancer tissues were gained, which were widely low expressed in carcinomas especially in nasopharyngeal carcinomas. Expression of Cx26, Cx32, Cx43, and Cx45 in carcinomas such as carcinomas of colon and recta, hepatocellular carcinomas, gastric carcinomas, lung carcinomas and thyroid cancers were significantly lower than their related adjacent-cancer tissues. CONCLUSION: Low expression of Cx26, Cx32, Cx43, and Cx45 might play crucial roles in carcinogenesis of many kinds of carcinomas. Tissue microarrays consisting of many different kinds of carcinomas provide a new high-throughout tool for rapid and comprehensive molecular expression profiles of carcinomas, such as connexin genes.