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Classical swine fever virus (CSFV), African swine fever virus (ASFV), and Erysipelothrix rhusiopathiae ( E. rhusiopathiae ) remain endemic in many parts of China. Co-infections make distinguishing their clinical symptoms and pathological changes difficult. This study developed a multiplex real-time quantitative reverse transcription polymerase chain reaction (multiplex qRT-PCR) that can simultaneously detect CSFV, ASFV, and E. rhusiopathiae . Three sets of primers and probes were designed to target the CSFV 5΄ untranslated region, ASFV p72 gene, and E. rhusiopathiae 16sRNA gene. Multiplex qRT-PCR for simultaneous differential detection of these three pathogens was developed after optimizing reaction parameters such as annealing temperature, primer and probe concentrations, amplification cycles, etc. The multiplex qRT-PCR could detect CSFV, ASFV, and E. rhusiopathiae simultaneously but could not amplify other porcine pathogens. The assay's limit of detection (LOD) was 2.89 × 10 2 copies/μL for CSFV, ASFV, and E. rhusiopathiae . All correlation coefficients (R 2 ) at higher than 0.99, and the amplification efficiency was 98, 90, and 84%, respectively. All correlation coefficients (R2) were higher than 0.99, and the efficacy of amplification was 84%. In a repeatability test utilizing standard recombinant plasmids, the intra- and inter-assay coefficients of variation (CVs) were less than 2.27 and 3.79 percent, respectively. Lastly, 150 clinical samples were used to evaluate the assay's applicability in the field. The positive rates of CSFV, ASFV, and E. rhusiopathiae were 1.33%, 0, and 3.33%, respectively. And no co-infection among the three pathogens was found. The concordance rate between the multiplex qRT-PCR and single-plex commercial PCR kits reached 100%. This study's multiplex qRT-PCR could provide a rapid, sensitive, and specific method for the simultaneous and differential detection of CSFV, ASFV, and E. rhusiopathiae ., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Zhao, Wen, Jia, Zhou, Luo, Lv and Zhai.)

Atherosclerosis is the major pathophysiological basis of a variety of cardiovascular diseases and has been recognized as a lipid-driven chronic inflammatory disease. Gelsolin (GSN) is a member of the GSN family. The main function of GSN is to cut and seal actin filaments to regulate the cytoskeleton and participate in a variety of biological functions, such as cell movement, morphological changes, metabolism, apoptosis and phagocytosis. Recently, more and more evidences have demonstrated that GSN is Closely related to atherosclerosis, involving lipid metabolism, inflammation, cell proliferation, migration and thrombosis. This article reviews the role of GSN in atherosclerosis from inflammation, apoptosis, angiogenesis and thrombosis., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Pigs are the main host of Seneca Valley virus (SVV), previously known as Senecavirus A (SVA). Pigs affected by SVV have vesicles in the nose, hooves, and limp and may cause death in some severe cases. Occasionally, SVV has also been detected in mice, houseflies, environmental equipment, and corridors in pig farms. Moreover, it was successfully isolated from mouse tissue samples. In this study, an SVV strain (SVA/GD/China/2018) was isolated from a buffalo with mouth ulcers in the Guangdong province of China using seven mammalian cell lines (including BHK-21, NA, PK-15, ST, Vero, Marc-145, and MDBK). The genome of SVA/GD/China/2018 consists of 7,276 nucleotides. Multiple-sequence alignment showed that SVA/GD/China/2018 shared the highest nucleotide similarity (99.1%) with one wild boar-origin SVV strain (Sichuan HS-01) from the Sichuan province of China. Genetic analysis revealed that SVA/GD/China/2018 clustered with those porcine-origin SVV strains. To the best of our knowledge, this is the first report of SVV infection in buffalo, which might expand the host range of the virus. Surveillance should be expanded, and clinical significance of SVV needs to be further evaluated in cattle., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Zhou, Liang, Si, Li, Chen, Cai, Lv, Wen, Zhai, Zhai, Liao and He.)

Bovine rhinitis B virus (BRBV) is an emerging viral species in the genus Aphthovirus , family Picornaviridae. Studies suggested that BRBV was considered a potential etiological agent of bovine respiratory disease complex (BRDC). BRBV has been reported in the United States, Sweden, Canada, Japan, and Mexico. However, little information of BRBV was available in China. In this study, we performed viral metagenomic analysis in a calf with respiratory disease. The results showed high abundance (3.85) of BRBV nucleotide and 248 mapped reads in calf samples. Online BLASTn analysis showed that three contigs of those had the highest nucleotide similarity (95%) with one Swedish BRBV isolate (BRBV_SWE1, GenBank accession no. KY432299). To identify the genome characterization of the Chinese BRBV isolate (designated CHN1), six couples of overlapping RT-PCR primers were designed according to genome sequences of BRBV_SWE1. Through gene cloning and splicing, we obtained the genome information of CHN1, possessing 7,465 nucleotides (46.6% G+C). Although CHN1 had the highest nucleotide similarity (95.1%) with BRBV_SWE1, one 11-nucleotide (ACATTTGTTGT) deletion occurred in the 5' untranslated region compared to SWE1. Phylogenetic analysis showed that CHN1 clustered together with BRBV_SWE1, and far from other BRBV isolates. This study recorded the first discovery of BRBV infection in China. Further investigation should be made in order to evaluate the infection status and epidemiological significance of BRBV in China., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Zhai, Xie, Zhai, Wen, Lv, Chen, Wang and Wei.)

Bovine rhinitis B virus (BRBV) has been frequently identified in cattle diagnosed with bovine respiratory disease complex (BRDC) in recent years, suggesting its potential contribution to BRDC. The goal of this study was to develop a TaqMan-based real-time quantitative RT-PCR assay for efficient BRBV detection. A pair of primers and a probe were designed based on the 3D gene of the BRBV genome. The assay was specific for BRBV and able to exclude bovine rhinitis A virus, foot-and-mouth disease virus and Senecavirus A. The limit of detection of the assay was 4.46 copies per reaction. A standard curve was plotted, with a coefficient of determination of 0.999 in the concentration range of 10 0 -10 8 copies/μl. The reproducibility of the assay was acceptable, with the standard deviations of cycle threshold values lower than 1.00 in both intra- and inter-assay. Of 200 samples collected from 150 head of cattle in recent years in China, 11% (22/200) of the samples tested positive in the assay, i.e., 4.6% (7/150) of the cattle were BRBV positive. This study provides an efficient diagnostic tool for the epidemiological investigations of BRBV., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Xie, Lv, Wen, Zhai, Luo, Wei, Chen and Zhai.)

Targeted proteomics has advantages over earlier conventional technologies for protein detection. We developed and validated an LC/MRM-MS-based targeted proteomic method combined with immunoaffinity precipitation for the enrichment and detection of low abundance chemerin isoforms in human biofluids. After tryptic digestion, each chemerin isoform was characterized by isoform-specific peptides, and the absolute quantification was achieved by using stable isotope-labeled peptides as internal standards. In serum, follicular fluid and synovial fluid, a total of 6 chemerin isoforms were identified and quantified, among which a novel natural isoform 153Q was discovered for the first time. The relative content of the six chemerin isoforms in human serum was 157S ≫ 156F ≫ 158K > 154F ≥ 155A > 153Q in the ratio of 25:17:5:2.5:2.2:1, respectively. The absolute contents were in the range of 88-3.5 ng/mL. This distribution remained consistent among the 3 biofluids analyzed. Total chemerin were found to be increased in both polycystic ovary syndrome (serum and follicular fluid) and rheumatoid arthritis (serum) patients. However, chemerin isoform analysis revealed that only 156F & 157S were increased in the former, while 155A, 156F & 157S were increased in the latter. This demonstrates the potential of this method in detailed characterization of changes in chemerin isoforms that may be of clinical relevance., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Enzootic nasal tumor virus (ENTV) has two types, ENTV-1 in sheep and ENTV-2 in goats, respectively. In China, the incidence of ENTV-2 related diseases has increased year by year. In this study, we reported an outbreak of ENTV-2 in a commercial goat farm in Qingyuan city, Guangdong province, southern China. A full-length genome of ENTV-2 (designated GDQY2017), with 7479 base pairs, was sequenced. Although GDQY2017 shared the highest nucleotide identity with a Chinese ENTV-2 isolate (ENTV-2CHN4, GenBank accession number KU258873), it possesses distinct genome characteristics undescribed, including a non-continuous 21-nucleotide insertion in the gag gene and a non-continuous 12-nucleotide deletion in the env gene. Notably, most of these indel nucleotide sequences were originated from a Chinese jaagsiekte sheep retrovirus (JSRV) isolate (GenBank accession number DQ838494). In the gag and env genes, GDQY2017 was phylogenetically related to those Chinese ENTV-2 isolates and a Chinese JSRV isolate (DQ838494). For GDQY2017-like viruses, more surveillance work should be made to explain their pathogenicity in goat herds. To our knowledge, this study represents the first to demonstrate the circulating pattern of ENTV-2 in Guangdong province, China, which will help to better understand the epidemiology and genetic diversity of ENTV-2.

Porcine circovirus (PCV) is one of the smallest known DNA viruses in mammals. At present, PCVs are divided into three species, PCV1, PCV2, and PCV3. PCV1 and PCV2 were found in the 1970s and the 1990s, respectively, whereas PCV3 was discovered recently in 2016. PCV1 does not cause diseases in pigs. However, PCV3, similar to PCV2, is reported to be associated with several swine diseases, including porcine dermatitis and nephropathy syndrome (PDNS) and reproductive failure. PCVs are very common in domestic pigs as well as wild boars. However, PCVs have been occasionally isolated from non-porcine animals, including ruminants (such as cattle, goats, wild chamois, and roe deers), rodents (such as NMRI mice, BALB/c mice, Black C57 mice, ICR mice, Mus musculus , and Rattus rattus ), canines (such as dogs, minks, foxes, and raccoon dogs), insects (such as flies, mosquitoes, and ticks), and shellfish. Moreover, PCVs are frequently reported in biological products, including human vaccines, animal vaccines, porcine-derived commercial pepsin products, and many cell lines. PCVs are also abundant in the environment, including water samples and air samples. Interestingly, PCV1 and/or PCV2 antibody or antigen has also been detected in sera, stool samples and respiratory swab samples of human, revealing zoonotic potential of PCVs. Thus, PCVs inhabit many types of reservoirs. In this review, we summarize the reservoirs of PCVs, and this information would be helpful in understanding the natural circulating status and possible cross-species transmission of PCVs., (Copyright © 2019 Zhai, Lu, Wei, Lv, Wen, Zhai, Chen, Sun and Xi.)

Background: In prenatal diagnosis, CMA has begun to emerge as a favorable alternative to karyotype analysis, but it could not identify balanced translocations, triploidies, inversion and heteromorphisms. Therefore, conventional cytogenetic and specific staining methods still play an important role in the work-up of chromosome anomaly. This study investigated the application of C-banding and AgNOR-staining techniques in prenatal diagnosis of chromosomal heteromorphisms and some structure abnormalities., Results: Among the 2970 samples, the incidence of chromosomal heteromorphisms was 8.79% (261/2970). The most frequent was found to be chromosome Y (2.93%, 87/2970), followed by chromosome 1 (1.65 %, 49/2970), 9 (1.52 %, 45/2970), 22 (0.77 %, 23/2970) and 15 (0.64 %, 19/2970). We compared the incidence of chromosomal heteromorphisms between recurrent spontaneous abortion (RSA) group and control group. The frequency of autosomal hetermorphisms in RSA group was 7.63% higher than that in control group (5.78%), while the frequency of Y chromosomal heteromorphisms was 4.76% lower than that in control group (5.71%). Here we summarized 4 representative cases, inv (1) (p12q24), psu dic (4;17) (p16.3;p13.3), r(X)(p11; q21) and an isodicentric bisatellited chromosome to illustrate the application of C-banding or AgNOR-staining, CMA or NGS was performed to detect CNVs if necessary., Conclusions: This study indicated that C-banding and AgNOR-staining were still effective complementary methods to identify chromosomal heteromorphisms and marker chromosomes or some structural rearrangements involving the centromere or acrocentric chromosomes. Our results suggested that there was no evidence for an association between chromosomal heteromorphisms and infertility or recurrent spontaneous abortions. Undoubtedly, sometimes we needed to combine the results of CMA or CNV-seq to comprehensively reflect the structure and aberration of chromosome segments. Thus, accurate karyotype reports and genetic counseling could be provided., Competing Interests: Competing interestsThe authors declare that they have no competing interests.

Plants sense photoperiod signals to confirm the optimal flowering time. Previous studies have shown that Cryptochrome2 ( CRY2 ) functions to promote floral transition in the long-day plant (LDP) Arabidopsis ; however, the function and molecular mechanism by which CRY2 regulates floral transition in short-day plants (SDPs) is still unclear. In this study, we identified a CRY2 homologous gene, ClCRY2 , from Chrysanthemum lavandulifolium , a typical SDP. The morphological changes in the C. lavandulifolium shoot apex and ClFTs expression analysis under SD conditions showed that adult C. lavandulifolium completed the developmental transition from vegetative growth to reproductive growth after eight SDs. Meanwhile, ClCRY2 mRNA exhibited an increasing trend from 0 to 8 d of SD treatment. ClCRY2 overexpression in wild-type (WT) Arabidopsis and C. lavandulifolium resulted in early flowering. The transcript levels of the CONSTANS-like ( COL ) genes ClCOL1 , ClCOL4 , and ClCOL5 , and FLOWERING LOCUS T ( FT ) homologous gene ClFT1 were upregulated in ClCRY2 overexpression ( ClCRY2- OE) C. lavandulifolium under SD conditions. The transcript levels of some circadian clock-related genes, including PSEUDO-REPONSE REGULATOR 5 ( PRR5 ), ZEITLUPE ( ZTL ), FLAVIN-BINDING KELCH REPEAT F-BOX 1 ( FKF1 ), and GIGANTEA ( GI-1 and GI-2 ), were upregulated in ClCRY2- OE C. lavandulifolium , while the expression levels of other circadian clock-related genes, such as EARLY FLOWERING 3 ( ELF3 ), ELF4 , LATE ELONGATED HYPOCOTYL ( LHY ), PRR73 , and REVEILLE8 ( RVE8 ), were downregulated in ClCRY2- OE C. lavandulifolium under SD conditions. Taken together, the results suggest that ClCRY2 promotes floral transition by fine-tuning the expression of circadian clock-related gene, ClCOL s and ClFT1 in C. lavandulifolium under SD conditions., Competing Interests: The authors declare that they have no conflict of interest.

Silicon (Si) plays important roles in alleviating heavy metal stress in rice plants. Here we investigated the physiological response of rice at different growth stages under the silicon-induced mitigation of cadmium (Cd) and zinc (Zn) toxicity. Si treatment increased the dry weight of shoots and roots and reduced the Cd and Zn concentrations in roots, stems, leaves and grains. Under the stress of exposure to Cd and Zn, photosynthetic parameters including the chlorophyll content and chlorophyll fluorescence decreased, while the membrane permeability and malondialdehyde (MDA) increased. Catalase (CAT) and peroxidase (POD) activities increased under heavy metals stress, but superoxide dismutase (SOD) activities decreased. The magnitude of these Cd- and Zn-induced changes was mitigated by Si-addition at different growth stages. The available Cd concentration increased in the soil but significantly decreased in the shoots, which suggested that Si treatment prevents Cd accumulation through internal mechanisms by limiting Cd 2+ uptake by the roots. Overall, the phenomena of Si-mediated alleviation of Cd and excess Zn toxicity in two rice cultivars could be due to the limitation of metal uptake and transport, resulting in an improvement in cell membrane integrity, photosynthetic performance and anti-oxidative enzyme activities after Si treatment.

Porcine reproductive and respiratory syndrome virus (PRRSV) is considered an important economic pathogen for the international swine industry. At present, both PRRSV-1 and PRRSV-2 have been confirmed to be co-circulating in China. However, there is little available information about the prevalence or distribution of PRRSV-1 in Guangdong province, southern China. In this study, we performed molecular detection of PRRSV-1 in 750 samples collected from 50 farms in 15 major pig farming regions in this province. After RT-PCR testing, 64% (32/50) of farms were confirmed as PRRSV-1-positive. Surprisingly, PRRSV-1 was circulating on at least one pig farm in all 15 regions; of the 750 samples, 186 samples (24.8%) were positive for PRRSV-1. Furthermore, 15 representative PRRSV-1 ORF5 sequences (606 bp) (n = 1 per region) were obtained from those PRRSV-1-positive regions. Sequence alignment analysis indicated that they shared 81.8% ~ 100% nucleotide and 81.2% ~ 100% amino acid similarity with each other. Although all current PRRSV-1 sequences were divided into pandemic subtype 1, most of them had unique glycoprotein-5 amino acid sequences that are significantly different from other known PRRSV-1 isolates. To conclude, the present findings revealed wide geographical distribution of PRRSV-1 in Guangdong province, southern China. This study further extends the epidemiological significance of PRRSV-1 in China.

An efficient transition-metal-free oxidative cyclization reaction using isatins and alkynes for the facile synthesis of structurally diverse 4-quinolones has been developed. Intriguingly, switchable access to substituted 3-carboxylate-4-quinolones and 1-vinyl-3-carboxylate-4-quinolones could be achieved by choosing a different base in the reaction. The obtained products could undergo further transformations, increasing the application potential of the method in organic synthesis.

Objective: Chromosome aberrations are generally considered as one of the most substantial causative factors contributing to spontaneous miscarriages. Cytogenetic analyses like G-banded karyotype and chromosomal microarray analyses are often performed to further investigate the chromosome status of a miscarried fetus., Study Design: Here, we describe a novel method, AnnoCNV, to detect DNA copy number variations (CNVs) using low coverage whole genome sequencing (WGS). We investigated the overall frequency of chromosomal abnormalities in 149 miscarriage specimens using AnnoCNV., Results: Among 149 fetal miscarriage samples, more than two fifths of them (42.95%, 64) carried at least one chromosomal abnormality, and a subset (40) was identified as autosomal trisomy which account for 26.84% of all samples. We have also developed a robust algorithm in AnnoCNV, which is able to differentiate specifically karyotype 69,XXY from sex chromosomal aneuploidy 45,X, and to identify 45,X/46,XX mosaicism. Lastly, across the whole genome AnnoCNV identifies CNVs, which are associated with both reported symptoms and unknown clinical conditions., Conclusion: This cost-effective strategy reveals genome wide discovery of chromosome aberrations at higher resolution, which are consistent with parallel investigation conducted by SNP based assay., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Schmallenberg virus (SBV) is an emerging and rampant arbovirus in Europe, and even Africa and West Asia. Investigating whether SBV existed in new regions or countries, it was very helpful for the early warning and control of SBV. In this study, we collected 317 serum samples (n = 242 for dairy cattle, n = 13 for yellow cattle, n = 21 for buffalo, and n = 41 for goats) from Guangdong province of southern China, which is located in a subtropical region and is an important distribution area for arboviral diseases. A commercial competitive enzyme-linked immunosorbent assay (cELISA) kit and a previously established real-time PCR were used to detect SBV antibody and RNA in those serum samples. Via testing, serological evidence of SBV was confirmed, with total positive rates (57.4, 15.4, 19, and 9.8%) in dairy cattle, yellow cattle, buffalo, and goats, respectively, while no positive signal for SBV RNA was found. To summarize, this study for the first time provided preliminary serological evidence of SBV infection in China, East Asia. Further investigations on molecular evidence, origin, and pathogenesis of SBV in ruminants needed to be studied in China.

Background: Porcine circovirus type 3 (PCV3), as an emerging circovirus species, was reported to be widely circulating in the United States, China, South Korea and Poland. Previous studies revealed that PCV3 was mainly concentrated in sick animals with respiratory disease, skin disease, reproductive disorders and so on. However, the circulating status of PCV3 in pigs with other clinical presentations (especilly asymptomatic or diarrhea) was not well established., Findings: In this study, to conduct a comparative epidemiological survey of PCV3, 80 weaned pig serum samples with severe respiratory disease (SRD), 175 weaned pig serum samples with mild respiratory disease (MRD), 216 asymptomatic weaned pig serum samples, 35 diarrheal weaned pig samples and 35 non-diarrheal weaned pig samples were collected from eight provinces of China. Via qPCR testing, PCV3 was circulating in all sampling provinces, with total positive rates varying from 1.04% to 100%. Interestingly, the PCV3-positive rate was significantly higher in weaned pigs with SRD (63.75%, 51/80) than in those weaned pigs with MRD (13.14%, 23/175) and asymptomatic pigs (1.85%, 4/216) (P < 0.01). Similarly, the PCV3-positive rate was significantly higher in diarrheal weaned pigs (17.14%, 6/35) than in non-diarrheal weaned pigs (2.86%, 1/35) (P < 0.05). Moreover, the lower Ct values of qPCR were frequently found in those weaned pigs or fattening pigs with respiratory disease and diarrhea rather than that in asymptomatic pigs. Sequence analysis showed that low genetic diversity existed among those PCV3 sequences collected from pigs with different clinical presentations., Conclusions: The present study further extends evidence that newly described PCV3 widely circulates in six additional provinces of Southern and Northern China and has high similarity to previously reported isolates. As an emerging virus of swine, although the present case-control study reveals that PCV3 has a potential association with swine respiratory disease and diarrhea, further investigations into the pathogenesis are needed to ascertain the role of PCV3 in swine health.

Molecular tests revealed influenza D viruses of D/OK lineage widely circulating in farmed animal species in Guangdong Province, southern China. In particular, we found high levels of influenza D virus infection in goats and pigs. We also detected viral RNA in serum specimens and feces of animals with certain severe diseases.

Here, we describe a novel porcine circovirus type 2 (PCV2) variant (GD2014) found in the Guangdong province, southern China. Its complete genome is 1,766 nucleotides and contained a 708-nucleotide open reading frame 2 (ORF2). Sequence analysis suggested that GD2014 is closest to JS2015 originating from the Jiangsu province of China and belongs to the PCV2d genotype., (Copyright © 2017 Zhai et al.)

Porcine circovirus type 2 (PCV2) is considered the major etiological pathogen of porcine circovirus-associated diseases (PCVADs) in pigs. Recently, PCV2 was also found in non-porcine animals such as cattle, rats, and mice. However, there was no record of PCV2 in rats in China. The goal of this study was to investigate whether PCV2 was present in rats (Rattus norvegicus, RN) on three swine farms, using molecular tools. PCR results showed that 30 of 95 (31.6 %) rat samples were positive for PCV2. Moreover, further genotype analysis suggested that 10 of 30 (33.3 %) were positive for PCV2a, 19 of 30 (63.3 %) were positive for PCV2b, and only one sample (1/30, 3.33 %) was co-infected by PCV2a and PCV2b. To determine the possible origin of PCV2, 60 serum samples were also collected from weaned pigs on those swine farms, and 23 out of 60 samples were positive for PCV2. In addition, two distinct RN-origin and two distinct porcine-origin PCV2 full-length nucleotide sequences were obtained from the farms. Sequence and phylogenetic analysis indicated that they had the highest nucleotide similarity and closest genetic relationships to each other. In this study, we report the infection and genome characterization of PCV2 in rats and compare RN-origin and porcine-origin PCV2 sequences obtained from the same pig farm, revealing possible cross-species transmission of PCV2.

Background: Following the initial isolation of porcine deltacoronavirus (PDCoV) from pigs with diarrheal disease in the United States in 2014, the virus has been detected on swine farms in some provinces of China. To date, little is known about the molecular epidemiology of PDCoV in southern China where major swine production is operated., Results: To investigate the prevalence of PDCoV in this region and compare its activity to other enteric disease of swine caused by porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis coronavirus (TGEV), and porcine rotavirus group C (Rota C), 390 fecal samples were collected from swine of various ages from 15 swine farms with reported diarrhea. Fecal samples were tested by reverse transcription-PCR (RT-PCR) that targeted PDCoV, PEDV, TGEV, and Rota C, respectively. PDCoV was detected exclusively from nursing piglets with an overall prevalence of approximate 1.28 % (5/390), not in suckling and fattening piglets. Interestingly, all of PDCoV-positive samples were from 2015 rather than 2012-2014. Despite a low detection rate, PDCoV emerged in each province/region of southern China. In addition, compared to TGEV (1.54 %, 5/390) or Rota C (1.28 %, 6/390), there were highly detection rates of PEDV (22.6 %, 88/390) in those samples. Notably, all five PDCoV-positive piglets were co-infected by PEDV. Furthermore, phylogenetic analysis of spike (S) and nucleocapsid (N) gene sequences of PDCoVs revealed that currently circulating PDCoVs in southern China were more closely related to other Chinese strains of PDCoVs than to those reported in United States, South Korea and Thailand., Conclusions: This study demonstrated that PDCoV was present in southern China despite the low prevalence, and supported an evolutionary theory of geographical clustering of PDCoVs.

Reticuloendotheliosis virus (REV), an important immunosuppressive pathogen, has many hosts, including chickens, ducks, geese, turkeys, and wild birds. Clinically, REV may lead to increased susceptibility to other pathogens, resulting in serious tissue damage (especially tumors) and the death of its host. In this study, we encountered a disease outbreak resulting in a large number of deaths of pigeons in Guangdong Province, Southern China. Histopathological analysis revealed apparent tumor-like lesions in multiple organs of pigeons. PCR assays for detection of tumor-associated pathogens (REV, avian leukosis virus, and Marek's disease virus) in poultry revealed the presence of REV sequences only. Moreover, fowlpox virus (FPV) with an insertion of REV long terminal repeat (LTR) sequences was also considered, but it was excluded using a specific PCR assay. To gain more genetic information, two full-length REV genome sequences were determined and found to have the highest nucleotide sequence similarity (99.9 %) and the closest genetic relationship to a vaccine strain (MD-2) and had a more distant genetic relationship (94.3 %) to a duck-origin strain (ATCC-VR775). To confirm the presence of REVs in pigeons, specific-pathogen-free (SPF) chickens and healthy pigeons were inoculated with microfiltered tumor tissue homogenates and were found to be susceptible to infection with REV. To our knowledge, this is the first report of REV in pigeons, and the data suggest that pigeons may be the natural host of REV.

Since 2013, the second outbreak of peste des petits ruminants (PPR) caused by Peste des petits ruminants virus (PPRV) has spread over more than 20 provinces, municipalities, and autonomous regions in China, resulting in major economic losses for livestock industry. In 2014, we encountered a clinical PPR case on a goat farm in Guangdong province, southern China. The complete genome of this PPRV strain, named CH/GDDG/2014, was sequenced to determine its similarities and differences with other strains. The CH/GDDG/2014 genome comprised 15,954 nucleotides (six nucleotides more than classical PPRVs identified before 2013, but complying with the rule of six) with six open reading frames encoding nucleocapsid protein, phosphoprotein, matrix protein, fusion protein, hemagglutinin, and large polymerase protein, respectively. The whole-genome-based alignment analysis indicated that CH/GDDG/2014 had the most proximate consensus (99.8 %) to China/XJYL/2013 and the least consensus (87.2 %) to KN5/2011. The phylogenetic analysis showed that CH/GDDG/2014 was clustered in one branch (lineage IV) with other emerging strains during the second outbreak. This study is the first report describing the whole-genome sequence of PPRV in Guangdong province, southern China and also suggests the PPR outbreak may be closely related to illegal cross-regional importation of goats.

For the worldwide pig industries, porcine circovirus type 2 (PCV2) is an economically important pathogen. At present, the prevalence of PCV2 is common in Chinese swine herds. However, there is little information on PCV2 prevalence in non-porcine animals in China, such as bovids. Therefore, the goal of this study is to obtain the firsthand prevalence data of PCV2 in bovids in China. Two hundred and eighty serum and muscle samples from dairy cows (n = 180), buffalo (n = 50), and yellow cattle (n = 50) were analyzed by PCR. The detection results show that PCV2 infections (16 %, 8/50) only exist in buffaloes. In addition, there are different PCV2 viral DNAs identified by differential PCR in the same buffalo sample. Nucleotide sequencing and phylogenetic analysis results based on partial ORF1 and ORF2 sequences suggest that PCV2 strains have genetic diversity in buffaloes and they are divided into three different genotypes (PCV2b, PCV2d, and PCV2e, respectively). Moreover, to our knowledge, the PCV2d and PCV2e genotypes have not been previously reported in bovids. Through this study, the firsthand data of PCV2 prevalence in bovids in China was documented.

Currently, porcine circovirus type 2 (PCV2) is considered the major pathogen of porcine circovirus associated-diseases (PCVAD) that causes large economic losses for the swine industry in the world annually, including China. Since the first report of PCV2 in 1998, it has been drawing tremendous attention for the government, farming enterprises, farmers, and veterinary practitioners. Chinese researchers have conducted a number of molecular epidemiological work on PCV2 by molecular approaches in the past several years, which has resulted in the identification of novel PCV2 genotypes and PCV2-like agents as well as the description of new prevalence patterns. Since late 2009, commercial PCV2 vaccines, including the subunit vaccines and inactivated vaccines, have already been used in Chinese swine farms. The aim of this review is to update the insights into the prevalence and control of PCV2 in China, which would contribute to understanding the epidemiology, control measures and design of novel vaccines for PCV2.

Ruminal impaction is considered an important internal disease in ruminants, such as dairy cows, sheep, and goats. It has been reported that its occurrence is associated with many different causes. This study describes a novel case of ruminal impaction caused by a plant, Ficus esquiroliana Levl., in Boer goats. This case suggests that Ficus esquiroliana should be taken into consideration when providing food for ruminants.

Objective: The aim of this study was to compare the basic characters of the speech-evoked auditory brainstem response (speech-ABR) between young adults and school-age children., Methods: Speech-ABR of thirty eight normal hearing subjects including eighteen school-age children and twenty young adults were recorded. The speech syllable/da/as stimulus sound was transmitted to right ear by insertion the earphones in speech-ABR test., Results: Response waves of speech-ABR in school-age children were similar to those in young adults, which contained the onset response (peak V and A), the transition (peak C), the frequency following responses (peak D, E and F) and the offset response (peak O). Both the latency and amplitude showed no significant difference in all waves between young adults and school-age children, except the latency of wave O and amplitude of wave F. The latency of O wave in school-age children (47.80 ± 0.38) ms were significantly shorter than that in adults (48.10 ± 0.40) ms (t = 2.330,P = 0.026). The amplitude of F wave in school-age children (-0.21 ± 0.15) µV were significantly larger than that in adults (-0.12 ± 0.08) µV (t = 2.146,P = 0.043)., Conclusion: Both the latency and amplitude of the speech ABR in school age children at 6 - 11 years old show the great similarity with the young adults, which indicate that the ability of speech processing of brainstem in children has completely reached maturity.

Objective: To assess the accuracy of a wrist-worn device (Watch-PAT 200) in the diagnosis of obstructive sleep apnea syndrome (OSAHS)., Methods: Forty-three adult subjects with suspected OSAHS simultaneously had a standard in-laboratory polysomnogram (PSG) and wore the Watch-PAT 200 during a full-night recording. PSG sleep and respiratory events were scored according to standard criteria. The PSG recordings were blindly manually analyzed, while Watch-PAT data were scored automatically based on the algorithm developed previously., Results: The mean age of the subjects was (42.2 ± 12.2) years (x(-) ± s), and mean body mass index was (28.0 ± 3.9) kg/m(2). Mean PSG apnea hypopnea index (AHI) was (34.9 ± 29.9) events per hour, and mean PAT-AHI was (36.0 ± 29.2) events per hour. There was a significant correlation between PAT AHI and AHI by PSG (r = 0.931, P < 0.01). A Bland-Altman plot of PAT AHI and PSG AHI was also used to assess the accuracy of Watch-PAT 200. At lower levels of AHI, PAT tended to overestimate disease severity, while at higher levels of AHI, Watch-PAT underestimated severity. To assess sensitivity and specificity of Watch-PAT, constructed receiver operator characteristic curves using a variety of AHI threshold values (5, 15 and 30 events per hour). For AHI ≥ 5 events per hour as threshold value, the Watch-PAT diagnosing rate was 93%, and sensitivity as well as specificity were 94.7% and 80.0%. The misdiagnosis rate and missed diagnosis rate were 20.0% and 5.3%. Optimal combinations of sensitivity and specificity for the AHI threshold values (15 and 30 events per hour) were 82.6% and 100.0%, 95.0% and 95.7% respectively., Conclusion: The Watch-PAT 200 may offer an accurate, robust, and reliable ambulatory method for the detection of OSAHS, with minimal patient discomfort.

Objective: To investigate the effective way to test 4-year-old children's ability of sound localization in the horizontal plane., Methods: Using minimum audible angle (MAA) measure procedure on the basis of conditioned play audiometry, sound localization test was conducted for 4-year-old children at 0 degrees , +/- 45 degrees , +/- 90 degrees , +/- 135 degrees and 180 degrees standard positions in the horizontal plane., Results: The outcome of sound localization test for 4-year-old children separately were: MAA (0 degrees ) = (3.80 +/- 0.71) degrees , MAA (-45 degrees ) = (7.70 +/- 1.27) degrees , MAA (45 degrees ) = (7.10 +/- 1.39) degrees , MAA (-90 degrees ) = (8.15 +/- 2.38) degrees , MAA (90 degrees ) = (7.61 +/- 2.47) degrees , MAA (-135 degrees ) = (8.85 +/- 2.70) degrees , MAA (135 degrees ) = (8.30 +/- 1.42) degrees , MAA (180 degrees ) = (5.20 +/- 1.27) degrees . The MAA of eight standard positions were less than 10 degrees , and the MAA (0 degrees ) was the smallest one., Conclusions: Our findings suggest that MAA test procedure on the basis of conditioned play audiometry could be used to evaluate the ability of sound localization in 4-year-old children.

Objective: To work out the elicitation plan, obtain the mismatch negativity (MMN) and get out the laboratory normal value as well as to study the influence to MMN from the deviation of auditory stimuli., Methods: Hearing test of the tone burst stimulation was performed on 21 healthy young volunteers according to oddball stimulation sequence. Each subject was performed two kinds of auditory stimuli including frequency deviant stimuli and intensity deviant stimuli, and of each one included three series of stimulation. MMN was gained by subtracting the ERP of deviant stimuli from the ERP of standard stimuli. The latency and amplitude of each MMN were recorded, and then the effect of the deviant extent for MMN was analyzed., Results: By this setup the MMN of normal young people was recorded and normal value of latency and amplitude of MMN were got. In the group of frequency deviant stimuli, the MMN latency [(155.81 +/- 29.08) ms], if the frequency was up to 2000 Hz, was shorter than that when the frequency deviance was 1000 Hz [(182.89 +/- 45.85) ms, (183.32 +/- 43.33) ms] (P = 0.033, 0.030); when the deviant extent were the same, the latency had no obvious difference if changing the frequency of the standard and deviant stimuli (P = 0.973); the MMN amplitude of three groups [(3.85 +/- 2.22) microV, (2.90 +/- 2.05) microV, (2.66 +/- 2.12) microV] had no obvious difference among them (P > 0.05). In the group of intensity deviant stimuli, the MMN latency [(157.04 +/- 34.87) ms], if the frequency was up to 20 dB, was shorter (P = 0.025, 0.017) than that when the intensity deviance was 10 dB [(184.46 +/- 38.05) ms, (186.24 +/- 42.36) ms]. When the deviant extent were the same, the latency had no obvious difference (P = 0.882) if changing the intensity of the standard and deviant stimuli but only group 4 and group 6 [(3.41 +/- 1.64) microV, (2.37 +/- 1.47) microV] were different in evidence (P = 0.031) while the others had no obvious difference (P = 0.524, 0.122)., Conclusions: MMN was only related to the difference between standard stimuli and deviant stimuli, but there was no relationship between MMN and the notice, which indicate that MMN could objectively reflect the capability of brain to detect the change of stimuli. MMN is the representation of brain high-level sensory function.

To investigate the relationship between the methylation status of imprinted gene H19 and the development of mice derived completely from Embryonic stem cells (ES) by tetraploid embryo complementation. The methylation status of two loci at 5'UTR region in imprinted gene H19 in normal adult control mice, 22 adult ES mice, 8 newborn dead ES mice and embryonic stem (ES) cells with different passage number were detected by using of methylation-sensitive restriction endonuclease-PCR technique. The results indicated that the methylation status of the imprinted gene H19 in adult ES mice were identical to that of adult control mice. However, some significant differences in the methylation status of the imprinted gene H19 were found among newborn dead ES mice, adult ES mice and normal adult control mice. Furthermore, the methylation status of the imprinted gene H19 in ES cells were probably different from that of adult ES mice and normal adult control mice.

Objective: To investigate the association of male infertility with single nucleotide polymorphisms of the autosomal DAZL gene., Methods: Semen samples were collected from infertile men (infertility group, n=144) and healthy fathers (fertility group, n=53) in the northeast of China, and the former was further divided into 5 subgroups according to the WHO diagnostic criteria for oligoasthenoteratozoospermia (1999) i.e., oligoasthenoteratozoospermia, oligoasthenozoospermia, asthenoteratozoospermia, asthenozoospermia and non-oligoasthenoteratozoospermia. PCR-RFLP analysis was used to detect the polymorphism of SNP260. The SNP386 polymorphism of the DAZL gene was analysed with a PCR-SSCP based method. Sequencing and statistical analyses were performed., Results: The SNP260 polymorphism of the DAZL gene was found in both the infertility and the control groups, with no significant difference in between (P > 0.05). There was a larger percentage of SNP260AG in the subgroup of oligoasthenoteratozoospermia than in others, and the SNP386 polymorphism of the DAZL gene was not found in any of the subgroups., Conclusion: The SNP260 and SNP386 polymorphisms of the DAZL gene are not associated with male infertility in the northeast of China. Further research needs to be done to clarify the association between SNP260AG and oligoasthenoteratozoospermia. SNP386 polymorphism may be restricted in some small area in Taiwan only. Therefore neither of them can be used as a molecular marker for the genetic diagnosis of male infertility in the northeast of China.

The aim of this study was to determine whether the number of passages affected the developmental pluripotency of embryonic stem (ES) cells as measured by the attainment of adult fertile mice derived from embryonic stem (ES) cell/tetraploid embryo complementation. Thirty-six newborns were produced by the aggregation of tetraploid embryos and hybrid ES cells after various numbers of passages. These newborns were entirely derived from ES cells as judged by microsatellite DNA, coat-color phenotype, and germline transmission. Although 15 survived to adulthood, 17 died of respiratory failure, and four were eaten by their foster mother. From the 15 mice that reached adulthood and that could reproduce, none arose from ES cells at passage level 15 or more. All 15 arose from cells at passages 3-11. Our results demonstrate that the number of passages affects the developmental pluripotency of ES cells.

Objective: To improve the precision of the traditional segmentation of echocardiogram, by suppressing the influence from inherent speckle noises in medical ultrasonic images., Method: An automatic segmentation method based on reconstructed morphology was proposed in this paper. First, the opening and closing operations by reconstruction were imposed to the ultrasonic image. Second, the top-hat operation was used to extract the bright and/or dark features and to find out the boundaries corresponding to these features, whereby implemented the automatic segmentation., Result: The segmented echocardiogram had less artificial boundaries resulted from speckle noise, and could accurately be extracted the artery and ventricle., Conclusion: The presented method can detect both dark and bright objects accurately, and the boundary has a fine continuity. In addition, the algorithm is also applicable to the extraction of sole bright/dark features, accordingly to reduce the complexity and time needed and to improve the accuracy.