Sheng-Nan Chen

The costimulatory molecule B7-H4 promote tumor progression and cell proliferation through translocating into nucleus

B7-H4, a member of B7 family, is a transmembrane protein and inhibits T-cells immunity. However, in a variety of tumor cells, B7-H4 was detected predominantly in intracellular compartments with unknown mechanism and functions. In this study, we analyzed B7-H4 expression and subcellular distribution by immunohistochemistry in renal cell carcinoma (RCC) tissues. B7-H4 protein was detected on the membrane, in the cytosol and/or in the nucleus in tumor tissues. The membrane and nuclear expression of B7-H4 was significantly correlated with the tumor stages of RCC. Moreover, the membrane localization of B7-H4 was inversely correlated with the intensity of tumor infiltrates lymphocyte (TILs), whereas no association was observed between nuclear expression of B7-H4 and the density of TILs status. We further identified that B7-H4 is a cytoplasmic-nuclear shuttling protein containing a functional nuclear localization sequence (NLS) motif. A point mutation of B7-H4 NLS motif blocked the leptomycin B -induced nuclear accumulation of B7-H4. HEK293 cells stably expressing B7-H4 NLS mutant exhibited more potent inhibition in T-cell proliferation and cytokine production through increasing its surface expression compared with wild-type B7-H4 transfected cells owing to their increased surface expression. Most importantly, overexpression of wild-type B7-H4 in HEK293 cells enhanced tumor cell proliferation in vitro and tumorigenicity in vivo, promoted G1/S phase transition. The regulation of cell cycle by wild-type B7-H4 was partialy due to upregulation of Cyclin D 1 and Cyclin E. A mutation of B7-H4 NLS motif abolished the B7-H4-mediated cell proliferation and cell cycle regulation. Furthermore, B7-H4 wild-type confers chemoresistance activity to RCC cell lines including Caki-1 and ACHN. Our study provides a new insight into the functional implication of B7-H4 in its subcellular localization.

Porcine circovirus type 2 in China: an update on and insights to its prevalence and control

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.

Co-existence of multiple strains of porcine circovirus type 2 in the same pig from China

Pigs are often co-infected by different viral strains from the same virus. Up to now, there are few reports about co-existence of different porcine circovirus type 2 (PCV2) strains in China. The aim of this study was to evaluate it in Chinese swine herds. 118 PCV2 positive DNAs isolated from diseased pigs identified by classic PCR were re-detected using a modified differential PCR assay. The results indicated that co-existence rates of PCV2 were 32.2% (38/118) in diseased pigs and 0% (0/41) in asymptomatic pigs. Four PCV2 complete genomes were cloned from two co-infected samples and their nucleotide (nt) identities were 95%-97.3%. The phylogenetic analysis showed that four PCV2 strains were divided into different genotypes, PCV2a, PCV2b, PCV2d and PCV2e, respectively. In addition, co-existence were not detected in 41 serum samples from healthy pigs but PCV2 single infection (31.7%, 13/41) existed. These data revealed that the co-existence of different strains of PCV2 might contribute to the development of more severe clinical symptoms for pigs. This is the first report confirming the co-existence of different PCV2 strains in Chinese swine herds. Meanwhile, this study could help us to understand new infection and prevalence forms of PCV2 clinically.

Complete genome characterization and phylogenetic analysis of three distinct buffalo-origin PCV2 isolates from China.

Complete genome characterization of porcine circovirus type 2 (PCV2) for bovid origins was still unclear in China. Therefore, in this study, PCV2 full-length genome of buffalo-origin was amplified and analyzed using PCR, DNAStar and MEGA 5.1. Genome size of three distinct PCV2 strains (buffalo1, buffalo2 and buffalo3) was 1767 bp (48.56% G+C), 1767 bp (48.67% G+C) and 1768 bp (48.08% G+C), respectively. At the nucleotide level, their identity varied from 95% to 96% for complete genome, from 97% to 97.8% for ORF1, and from 90.6% to 94.4% for ORF2. At the amino acid level, their identity varied from 98.7% to 99% for ORF1, and from 88% to 94.9% for ORF2. Online Blast analysis showed that buffalo1, buffalo2 and buffalo3 had highest nucleotide identity (varied from 99.77% to 99.83%) with porcine-origin PCV2 strains. Moreover, in the phylogenetic tree, they were divided into three different clusters and belonged to the worldwide accepted genotypes of PCV2b, PCV2c and PCV2a, respectively. To summarize, this study first recorded complete genome information of PCV2 for non-porcine origins in China.

Influenza D Virus in Animal Species in Guangdong Province, Southern China

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.

Erratum to: First molecular detection of porcine circovirus type 2 in bovids in China

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.

Sequence Comparison and Phylogenetic Analysis of Several EmergingPorcine Bocaviruses and a Proposal Regarding Nomenclature

The first Bocavirus (bovine parvovirus 1, BPV1) was discovered in calves with diarrhea in 1961. Since then, a series of bocaviruses have emerged in many animals including dogs, humans, swine, gorillas, California sea lions, and cats. These viruses are associated with digestive tract and respiratory tract diseases. Several emerging porcine bocaviruses (PBoVs) have been reported on pig farms in the past four years. However, the nomenclature used to describe these viruses has been confused and irregular. The goal of this study was to compare the sequences of these emerging PBoVs, perform phylogenetic analysis, and present a proposal regarding their nomenclature. Genome sequences of some PBoVs and other bocaviruses were downloaded from the GenBank database. Sequence similarity, sequence mutation, deletion, or insertion, and sequence recombination among reference bocaviruses were obtained using DNAStar software, and its genetic evolution was determined using MEGA 5.1 software. According to the differences in their sequences and genome structure, the nomenclature of PBoVs was proposed and they were divided into four different species (from PBoV1 to PBoV4) and many different genotypes. Among them, both PBoV1 and PBoV2 had two genotypes, PBoV1-a and PBoV1-b and PBoV2-a and PBoV2-a and PBoV2-b, respectively. PBoV3 included three genotypes, PBoV3-a, PBoV3-b and PBoV3-c, respectively. In addition, for PBoV4, there were at least eight genotypes (PBoV4-a to PBoV4-h) in intraspecies. In conclusion, PBoVs were highly genetically diversified. The proposed nomenclature of PBoVs was used in the present studies, it could help us to better understand and standardize the nomenclature of PBoVs.

Diversified Strains of Porcine Reproductive and Respiratory Syndrome Virus Circulating in China: A Mini-overview

Porcine reproductive and respiratory syndrome virus (PRRSV) was considered as an important pathogen, which caused huge economic losses for the world swine industry annually. Until now, according to antigenic and genetic characteristics, two genotypes were identified, European (EU genotype, or type 1) genotype and North American genotype (NA genotype, or type 2), respectively. In China, both of them co-existed in swine herds, and even some novel viral strains emerged in the lastest years. The aim of the review was to describe genetic diversity of PRRSV based on non-structural protein 2 (Nsp2) in China, which could help us better understand molecular