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African swine fever (ASF) is an acute and highly contagious lethal infectious disease in swine that severely threatens the global pig industry. At present, a safe and efficacious vaccine is urgently required to prevent and control the disease. In this study, we evaluated the safety and immunogenicity of replication-incompetent type-2 adenoviruses carrying African swine fever virus (ASFV) antigens, namely CP204L (p30), E183L (p54), EP402R (CD2v), B646L (p72), and B602L (p72 chaperone). A vaccine cocktail delivered by simultaneous intramuscular (IM) and intranasal (IN) administration robustly elicited both systemic and mucosal immune responses against AFSV in mice and swine and provided highly effective protection against the circulating ASFV strain in farmed pigs. This multi-antigen cocktail vaccine was well tolerated in the vaccinated animals. No significant interference among antigens was observed. The combined IM and IN vaccination using this adenovirus-vectored antigen cocktail vaccine warrants further evaluation for providing safe and effective protection against ASFV infection and transmission.

Background: Gastrointestinal nematodes (GINs) have seriously affected the production and earnings of animal husbandry in various countries, while some species of GINs infect humans. At present, little is known about the species and prevalence of GINs in Qinghai-Tibetan Plateau (QTP)., Methods: In this study, 528 fresh faecal samples were collected from typical areas in different altitudes with seven species of livestock in Qinghai, Tibet, Gansu and Yunnan Provinces. ITS-2 rRNA gene of nematodes was employed to detect by PCR and sequencing analysis. Phylogenetic analysis of related sequences was performed using MEGA 6.0 software., Results: The overall prevalence of GINs was 80.3% with 20 species of GINs detected, while Teladorsagia circumcincta was the dominant one, and four of which were zoonotic species such as Trichostrongylus colubriformis, Trichostrongylus axei, Trichostrongylus vitrinus and Oesophagostomum stephanostomum., Conclusion: The study provided panoptic information on the prevalence and species diversity of GINs in QTP area, which is useful and valuable for reference of measure formulation in livestock husbandry and public health concerns. The parasites of T. circumcincta, Cylicocyclus nassatus, Strongylus edentatus, Cylicostephanus longibursatus, Telephlebia brevicauda, Cyathostomum catinatum, Mecistocirrus digitatus, Cooperia punctata, Cylicodontophorus bicoronatus, Nematodirus oiratianus and Oesophagostomum asperum were firstly reported the presence in QTP area. The study also showed that horse could be infected by T. circumcincta, goat could be infected by C. nassatus, cattle could be infected by S. edentatus and C. bicoronatus,and O. stephanostomum could infect yak, cattle and Mongolian sheep in worldwide. Nevertheless, more investigations are needed, such as microscopic examination, to accurately determine the species in the region., (© 2021 The Authors. Veterinary Medicine and Science published by John Wiley & Sons Ltd.)

Objective: Our previous studies have demonstrated that Plasmodium immunotherapy (infection) has antitumor effects in mice. However, as a new form of immunotherapy, this therapy has a weakness: its specific killing effect on tumor cells is relatively weak. Therefore, we tested whether Plasmodium immunotherapy combined with gemcitabine (Gem), a representative chemotherapy drug, has synergistic antitumor effects., Methods: We designed subcutaneously and intravenously implanted murine Lewis lung cancer (LLC) models to test the antitumor effect of Plasmodium chabaudi ASS (Pc) infection in combination with Gem treatment and explored its underlying mechanisms., Results: We found that both Pc infection alone and Gem treatment alone significantly inhibited tumor growth in the subcutaneous model, and combination therapy was more effective than either monotherapy. Monotherapy only tended to prolong the survival of tumor-bearing mice, while the combination therapy significantly extended the survival of mice, indicating a significant synergistic effect of the combination. In the mechanistic experiments, we found that the combination therapy significantly upregulated E-cadherin and downregulated Snail protein expression levels, thus inhibiting epithelial-mesenchymal transition (EMT) of tumor cells, which may be due to the blockade of CXCR2/TGF-β-mediated PI3K/Akt/GSK-3β signaling pathway., Conclusion: The combination of Pc and Gem plays a synergistic role in inhibiting tumor growth and metastasis, and prolonging mice survival in murine lung cancer models. These effects are partially attributed to the inhibition of EMT of tumor cells, which is potentially due to the blockade of CXCR2/TGF-β-mediated PI3K/Akt/GSK-3β/Snail signaling pathway. The clinical transformation of Plasmodium immunotherapy combined with Gem for lung cancer is worthy of expectation., Competing Interests: Authors ZT, WD, SZ, WH, FZ, and XPC were employed by CAS-Lamvac Guangzhou Biomedical Technology Co., Ltd. The remaining 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 Chen, Tao, Liang, Ma, Adah, Ding, Chen, Li, Dai, Fanuel, Zhao, Hu, Wu, Duan, Zhou, Qin, Chen and Yang.)

Twenty-four Tan sheep were randomly assigned into 4 groups to study the capability of tannin supplementation (0.5% in dietary DM) to lower blood lipid levels mediated through the gut microbiota. The control (NC) group was offered a basic diet, while the 3 treatment groups were the TA group, which received supplementary tannic acid (TA); GSPE group, which received supplementary procyanidins (GSPE); and the TA + GSPE group, which received supplementary TA and GSPE, besides being supplied with the basic diet for 8 weeks feeding. At the end of the experiment, the serum glucose, insulin, lipids, and cytokines were measured, and the short-chain fatty acids (SCFAs) in the colon were tested by GC/MS. Moreover, the jejunal and colonic microbiota were detected by 16S rRNA sequencing. Significant reductions in serum triacylglycerol, cholesterol, and high density lipoprotein were found in all treatments. The total SCFAs decreased, while the iso-acids were significantly increased in the TA and TA + GSPE groups. The sheep showed noticeably lower MCP-1 and higher COX-2 levels in the GSPE group than that in the NC group. IL-6 was increased in the sheep fed with TA. The tannins still caused a noticeable shift in the colonic microbiota, with significant increases in the abundances of Adlercreutzia and Oscillospira . Ultimately, it was found that the diet with low levels of tannin could reduce blood triacylglycerol and cholesterol in sheep significantly by affecting the composition of the gut microbiota.

Interscapular brown adipose tissue (iBAT) of both rabbits and humans exhibits a similar whitening phenomenon under physiological conditions. However, a detailed characterization of iBAT whitening in them is still lacking. Here, we chose rabbits as a model to gain a better understanding of the molecular signature changes during the whitening process of iBAT by transcriptomic analysis of rabbit iBAT at day 1, day 14, 1 month and 4 months after birth. We applied non-invasive MRI imaging to monitor the whitening process and correlated these changes with analysis of morphological, histological and molecular features. Principal component analysis (PCA) of differentially expressed genes delineated three major phases for the whitening process as Brown, Transition and Whitened BAT phases. RNA-sequencing data revealed that whitening of iBAT was an orchestrated process where multiple types of cells and tissues participated in a variety of physiological processes including neovascularization, formation of new nervous networks and immune regulation. Several key metabolic and signalling pathways contributed to whitening of iBAT, and immune cells and immune regulation appeared to play an overarching role.

Immune checkpoint blockade therapy (ICB) is ineffective against cold tumors and, although it is effective against some hot tumors, drug resistance can occur. We have developed a Plasmodium immunotherapy (PI) that can overcome these shortcomings. However, the specific killing effect of PI on tumor cells is relatively weak. Radiotherapy (RT) is known to have strong specific lethality to tumor cells. Therefore, we hypothesized that PI combined with RT could produce synergistic antitumor effects. We tested our hypothesis using orthotopic and subcutaneous models of mouse glioma (GL261, a cold tumor) and a subcutaneous model of mouse non-small cell lung cancer (NSCLC, LLC, a hot tumor). Our results showed that, compared with each monotherapy, the combination therapy more significantly inhibited tumor growth and extended the life span of tumor-bearing mice. More importantly, the combination therapy could cure approximately 70 percent of glioma. By analyzing the immune profile of the tumor tissues, we found that the combination therapy was more effective in upregulating the perforin-expressing effector CD8 + T cells and downregulating the myeloid-derived suppressor cells (MDSCs), and was thus more effective in the treatment of cancer. The clinical transformation of PI combined with RT in the treatment of solid tumors, especially glioma, is worthy of expectation.

The gut microbiota is closely associated with the health and production performance of livestock. Partial studies on ruminant microbiota are already in progress in the Qinghai-Tibetan Plateau Area (QTPA) in China, but large-scale and representative profiles for the QTPA are still lacking. Here, 16S rRNA sequencing was used to analyze 340 samples from yak, cattle, yak-cattle hybrids, and Tibetan sheep, which lived in a shared environment from 4 eco-regions of the QTPA during the same season, and aimed to investigate the fecal microbiota community composition, diversity, and potential function. All samples were clustered into 2 enterotypes, which were derived from the genera Ruminococcaceae UCG-005 and Acinetobacter, respectively. Environment, human activity, species, and parasitization all affected the fecal microbiota. By assessing the relationship between the fecal microbiota and the above variables, we identified a scattered pattern of fecal microbiota dissimilarity based more significantly on diet over other factors. Additionally, gastrointestinal nematode infection could reduce the capacity of the bacterial community for biosynthesis of other secondary metabolites, carbohydrate metabolism, and nucleotide metabolism. Ultimately, this study provided a fecal microbiota profile for ruminants living in 4 eco-regions of the QTPA and its potential future applications in developing animal husbandry regimes. IMPORTANCE Cattle, yak, and sheep reside as the main ruminants distributed throughout most regions of Qinghai-Tibetan Plateau Area (QTPA) in China. However, there is a lack of large-scale research in the QTPA on their fecal microbiota, which can regulate and reflect host health as an internalized "microbial organ." Our study depicted the fecal microbiota community composition and diversity of yak, cattle, yak-cattle hybrids, and Tibetan sheep from 4 eco-regions of the QTPA. Additionally, our results demonstrated here that the ruminant samples could be clustered into 2 enterotypes and that diet outweighed other factors in shaping fecal microbiota in the QTPA. This study provided a basis for understanding the microbiota characteristic of ruminants and its possible applications for livestock production in the QTPA.

Objective: The objective of this study was to investigate the effect of 4-month intensive feeding on the meat quality, fatty acid profile, flavor, and growth performance of grazing Hulunbuir sheep (HBS)., Methods: The HBS were selected 4-months after birth in a pasture rearing system as the experimental animals (n = 44, female, average body weight 23.8±2.2 kg) then divided equally into pasture-based grazing fattening (PAS) and concentrate-included intensive fattening (CON) groups for another 4-month finishing. When finished fattening, all animals were slaughtered to collect musculus longissimus dorsi subcutaneous adipose tissue and to investigate the influences on meat quality, fatty acid profile, flavor and growth performance., Results: The results showed lambs in CON group got significantly higher live weight, hot carcass weight, and dressing percentage. The CON group had significantly higher value of redness (a*), lightness (L*) and water holding capacity (p<0.05), significantly lower value of Warner-Bratzler shear force than the PAS group (p<0.05). The subcutaneous fat from CON group lambs demonstrated a significantly higher content of C18:1 and C18:2 (p<0.05), but lower C14:0 and C16:0, indicating an increased degree of unsaturated fatty acid. The content of 4-methyloctanoic acid, 4-ethyloctanoic acid and 4-methylnonanoic acid had increased 2 to 4 times, representing a more intense odor in the CON group. However, the values were still lower than most sheep breeds reported, indicating the indoor feeding system could not fundamentally deteriorate the excellent meat characteristic of HBS., Conclusion: It was evident that lambs in CON group exhibited a better meat production performance, improved in meat color, texture and healthier fatty acid profile through pastureweaned concentrate included intensive fattening system, which offers a good alternative regimen for lamb finishing and has a wide prospection in the HBS meat industry.

Zoo-archaeological and genetic evidence suggest that pigs were domesticated independently in Central China and Eastern Anatolia along with the development of agricultural communities and civilizations. However, the genetic history of domestic pigs, especially in China, has not been fully explored. In this study, we generate 42 complete mitochondrial DNA sequences from ∼7500- to 2750-year-old individuals from the Yellow River basin. Our results show that the maternal genetic continuity of East Asian domestic pigs dates back to at least the Early to Middle Neolithic. In contrast, the Near Eastern ancestry in European domestic pigs saw a near-complete genomic replacement by the European wild boar. The majority of East Asian domestic pigs share close haplotypes, and the most recent common ancestor of most branches dates back to less than 20,000 years before present, inferred using new substitution rates of whole mitogenomes or combined protein-coding regions. Two major population expansion events of East Asian domestic pigs coincided with changes in climate, widespread adoption of introduced crops, and the development of agrarian societies. These findings add to our understanding of the maternal genetic composition and help to complete the picture of domestic pig evolutionary history in East Asia., Competing Interests: Conflict of interest The authors declare that they have no conflict of interest., (Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The aim of this study was to verify that Caragana korshinskii Kom. (CK) as a component of sheep forage influences lamb digestibility and rumen fermentation by altering the rumen microbial community. Hence, 12 female Tan sheep were allocated into 2 groups: receiving (CK group) or not (control group) 10% of the diet forage fraction with CK. During the 60-day experiment, growth performance, apparent digestibility, rumen volatile fatty acids (VFAs), and nitrogen balance were measured. Meanwhile, the rumen bacterial community diversity and composition were detected by the 16S rRNA sequence. The results indicated that the apparent digestibility of acid detergent fibre (ADF) tended to be higher (0.05 < P < 0.10), and the feed conversion efficiency was improved (P < 0.05) when CK was offered. Compared to those under alfalfa, the composition and abundance of the rumen microbial community were altered in the CK group, and the phylum Firmicutes, which is involved in promoting fibre digestion, increased in abundance. Moreover, VFAs tended to decrease (0.05 < P < 0.10), and the molar proportion of butyrate declined; similarly, levels of hypoxanthine and xanthine were lower (P < 0.05) in the sheep fed CK and may have been responsible for the decreased abundance of Fibrobacter spp., which are cellulolytic ruminal bacteria associated with VFA production., (© 2022. The Author(s).)

Myogenic factor 5 plays active roles in the regulation of myogenesis. The aim of this study is to expose the genetic variants of the MYF5 and its association with growth performance and ultrasound traits in grassland short-tailed sheep (GSTS) in China. The combination technique of sequencing and SNaPshot revealed seven SNPs in ovine MYF5 from 533 adult individuals (male 103 and female 430), four of which are novel ones located at g.6838G > A, g.6989 G > T, g.7117 C > A in the promoter region and g.9471 T > G in the second intron, respectively. Genetic diversity indexes showed the seven SNPs in low or intermediate level, but each of them conformed HWE (p > 0.05) in genotypic frequencies. Association analysis indicated that g.6838G > A, g.7117 C > A, g.8371 T > C, g.9471 T > G, and g.10044 C > T had significant effects on growth performance and ultrasound traits. The diplotypes of H1H3 and H2H3 had higher body weight and greater body size, and haplotype H3 had better performance on meat production than the others. In addition, the dual-luciferase reporter assay showed that there are two active regions in the MYF5 promoter located at −1799~−1197 bp and −514~−241 bp, respectively, but g.6838G > A and g.7117 C > A were out of the region, suggesting these two SNPs influence the phenotype by other pathway. The results suggest that the MYF5 gene might be applied as a promising candidate of functional genetic marker in GSTS breeding.

Three sampling strategies with a 16s rRNA high-throughput sequencing and gene expression assay (by RT-PCR) were designed, to better understand the host and probiotics effect on gut microbiota in sheep. Sampling: (1) colon contents and back-fat tissues from small-tailed Han sheep (SHS), big-tailed Hulun Buir sheep (BHBS), and short-tailed Steppe sheep (SHBS) ( n = 12, 14, 12); (2) jejunum, cecum and colon contents, and feces from Tan sheep (TS, n = 6); (3) feces from TS at 4 time points (nonfeeding, 30 and 60 feeding days, and stop feeding 30 days) with probiotics supplementation ( n = 7). The results indicated SHS had the highest Firmicutes abundance, the thinnest back-fat, and the lowest expression of C/EBPβ , C/EBPδ , ATGL , CFD , and SREBP1 . Some bacteria orders and families could be potential biomarkers for sheep breeds with a distinct distribution of bacterial abundance, implying the host genotype is predominant in shaping unique microbiota under a shared environment. The microbiota diversity and Bifidobacterial populations significantly changed after 60 days of feeding but restored to its initial state, with mostly colonies, after 30 days ceased. The microbiota composition was greatly different between the small and large intestines, but somewhat different between the large intestine and feces; feces may be reliable for studying large intestinal microbiota in ruminants.

Protozoans of Entamoeba spp. are globally distributed zoonotic parasites that infect diverse animal hosts and humans. Prevalence and species/genotypes distribution of Entamoeba spp. in domestic animals are not fully investigated on Qinghai-Tibetan Plateau (QTP), an animal husbandry and agriculture region of China. In a survey, 528 fecal samples were collected from 7 species of domestic animals on multiple locations across QTP region and analyzed by PCR and sequencing analysis. The overall prevalence of Entamoeba spp. infection in all examined animals was 97.9 %. Four Entamoeba species, E. bovis, E. moshkovskii, E. ecuadoriensis and E. histolytica were found, and majority (94.2 %) of Entamoeba-infected animals harbored E. bovis. Six Entamoeba genotypes, Entamoeba ribosomal lineages (RL) 1, 2, 3, 4, 8 and 9 were identified by sequencing analysis. Two zoonotic species, E. moshkovskii and E. histolytica, were present in horses, while E. ecuadoriensis and E. bovis were found in horses and all species of seven farm animals, respectively. It was also observed that six Entamoeba genotypes were distributed in animals in specific pattern. The results revealed high prevalence of Entamoeba spp. infection in livestock, broad range of hosts as well as diversity and species/genotype distribution of Entamoeba spp. in farm animals inhabiting on the high altitude QTP region., (Copyright © 2020 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Blastocystis is a common intestinal protozoan parasite of humans and a variety of animal species. To date, the prevalence of Blastocystis and major subtypes distribution in the domestic animals inhabiting in the Qinghai-Tibetan Plateau Area (QTPA) of China is yet poorly studied. In this study, we investigated the distribution and genetic diversity of Blastocystis in seven animal species in QTPA in China. Four hundred and five fresh fecal samples were collected from domestic animals in Qinghai, Yunnan, and Tibet of China and analyzed using nested PCR and SSU rRNA gene sequencing. It was found that the overall prevalence of Blastocystis infection was 40.2% (163/405) in the animals studied. The most predominant subtype of Blastocystis was ST10 (57.7%) followed by ST14 (28.8%) and ST2 (13.5%). These results reveal the epidemiological features of Blastocytis infection in animals in the high altitude plateau area. The finding of presence of ST2 in a number of animal species suggests a zoonotic nature of Blastocystis and might be of importance of public health., (Copyright © 2020. Published by Elsevier B.V.)

Accumulating evidence so far has shown that EBV's miRNAs have been found to be involved in cancer progression. However, the comprehensive EBV miRNA expression profiles and their biological significance in EBV-associated diseases are not well documented. A comprehensive profiling of EBV-encoded miRNAs expressed in CAEBV, EBV-HLH, and nasopharyngeal carcinoma (NPC) patients was constructed, and the results showed that miR-BART19-3p was upregulated in all these diseases. Ectopic expression of miR-BART19-3p induced EBV-negative cell proliferation and suppressed cell apoptosis. Molecularly, adenomatous polyposis coli (APC) was identified to be a direct target of miR-BART19-3p, and APC mRNA expression was inversely correlated with miR-BART19-3p in CAEBV samples. Our results demonstrated that miR-BART19-3p contributes to the tumorigenesis of EBV-associated diseases and may be a potential therapeutic target., Competing Interests: The authors report no conflicts of interest in this work., (Copyright © 2020 Qingxun Zhang et al.)

Cryptosporidium spp., Enterocytozoon bieneusi, and Giardia duodenalis are globally ubiquitous infectious parasitoses in humans and animals. The purpose of the present study was to determine the distribution and genetic diversity of Cryptosporidium spp., Enterocytozoon bieneusi, and Giardia duodenalis from animal sources in different regions of QTPA in China. A total of 405 fresh fecal specimens from seven domestic animals were collected in Qinghai, Yunnan, and Tibet. The overall prevalence of Cryptosporidium spp., E. bieneusi and G. duodenalis was 3.0% (n = 12), 19.8% (n = 80) and 5.7% (n = 23), respectively. Four Cryptosporidium species (C. andersoni, C. xiaoi, C. bovis, and C. ryanae) were identified and C. bovis was firstly identified from camel. Ten genotypes of E. bieneusi were identified in the present study, including eight known genotype (BEB6, CAM2, CHG2, CAM1, COS I, J, CHS8, and CHG3), and two novel genotypes (YAK1 and PN). Genotype CAM2 was detected in horse, yak, and mongolian sheep for the first time and the novel genotype YAK1 was grouped into the human-pathogenic group 1. G. duodenalis assemblage E was common in all animal species, and the human-pathogenic assemblage A was only detected in yaks and camels. Our results elucidate the occurrence and genetic diversity of three zoonotic pathogens from different animals and regions in QTPA, which could act as potential zoonotic reservoirs. More areas and larger number of samples are required to assess the potential risk of cross-species transmission in this region., (Copyright © 2019. Published by Elsevier Ltd.)

RIG-I signaling is critical to host innate immune response against RNA virus infection, and also can be activated against many kinds of cancer. Oncogene LMP1 of Epstein-Barr virus (EBV) contributes to various tumors progress. In this study, we have provided strong evidence that LMP1 inhibits Sendai virus mediated type I interferon production and downregulates RIG-I signaling pathway by promotion RIG-I degradation dependent on proteasome. Nineteen kinds of E3 ligase are identified by IP-MS as LMP1-interactors, they are candidate E3s, which are possibly recruited by LMP1 to mediate RIG-I degradation. CHIP is among these E3s, which has been reported to lead RIG-I degradation. Notably, we find C666-1, an EBV-positive nasopharyngeal carcinoma cell line, expresses low level of RIG-I, even treated with IFN-α, RIG-I expression could not be induced. This evidence indicates that EBV employs a unique strategy to evade RIG-I mediated immune responses.

A number of methods have been described to establish NK/T cell lines from patients with lymphoma or lymphoproliferative syndrome. These methods employed feeder cells, purified NK or T cells with as much as 10 mL of blood, or a high-dose of IL-2. This study presents a new method with a powerful and simple strategy to establish NK and T cell lines by culturing the peripheral blood mononuclear cells (PBMC) with the addition of recombinant human IL-2 (rhIL-2), and uses as little as 2 mL of whole blood. The cells can proliferate quickly in two weeks and be maintained for more than 3 months. With this method, 7 NK or T cell lines have been established with a high success rate. This method is simple, reliable, and applicable to establishing cell lines from more cases of CAEBV or NK/T cell lymphoma.

Porcine butyrophilin-like 5 (Btnl5) is a novel member of the butyrophilin family, which consists of immune regulators. The expression pattern and the function of this gene remain unclear. In this study, Btnl5 is identified as a negative regulator of the NF-κB pathway. Our study indicates that Btnl5 is mainly expressed in intestinal epithelial cells (IECs) and expressed in membrane systems. Btnl5 inhibits MyD88-mediated activation of the NF-κB pathway. Btnl5 interacts with TNF receptor-associated factor 2 (TRAF2) and transcription factor p65. Besides, Btnl5 inhibits p65-mediated activation of the NF-κB pathway and inhibits nuclear translocation of p65. These results suggest that Btnl5 may inhibit NF-κB pathway through binding and interfere nuclear translocation of p65., (Copyright © 2017. Published by Elsevier B.V.)

The influenza virus H1N1 has been prevalent all over the world for nearly a century. Many studies on its evolutionary history, substitution rate and antigenicity-associated sites have been done with small datasets. To have a complete view, we analysed 3171 full-length HA sequences from human H1N1 viruses sampled from 1918 to 2016, and discovered a new clade has formed with sequences isolated in Iran. Based on genetic distance calculations, we revealed an uneven evolutionary rate among sequences isolated in different years. We also found that the HA1 fragment of the new clade is like that of viruses that existed in the 1930s, while the HA2 fragment is closely associated with strains isolated after the 2009 pandemic. This new, "mixed" HA sequence indicates a cryptic antigenic shift event occurred, and it should draw more attention to the new clade identified from sequences from Iran., (Copyright © 2017. Published by Elsevier B.V.)

RIG-I is a key cytosolic pattern recognition receptor that interacts with MAVS to induce type I interferons (IFNs) against RNA virus infection. In this study, we found that cyclophilin A (CypA), a peptidyl-prolyl cis/trans isomerase, functioned as a critical positive regulator of RIG-I-mediated antiviral immune responses. Deficiency of CypA impaired RIG-I-mediated type I IFN production and promoted viral replication in human cells and mice. Upon Sendai virus infection, CypA increased the interaction between RIG-I and its E3 ubiquitin ligase TRIM25, leading to enhanced TRIM25-mediated K63-linked ubiquitination of RIG-I that facilitated recruitment of RIG-I to MAVS. In addition, CypA and TRIM25 competitively interacted with MAVS, thereby inhibiting TRIM25-induced K48-linked ubiquitination of MAVS. Taken together, our findings reveal an essential role of CypA in boosting RIG-I-mediated antiviral immune responses by controlling the ubiquitination of RIG-I and MAVS.

A number of methods exist for the transformation of B lymphocytes by the Epstein Barr virus in vitro into immortalized cell lines. We have developed a new method with a powerful and simple strategy for the establishment of B-LCLs, the red blood cell lysis method. This method simplified the PBMC separation procedure with red blood cell removal, and used as little as 0.5 mL of whole blood for establishing EBV-immortalized cell lines, which can proliferate to large cell numbers in a relatively short amount time with a 100% success rate. The method is simple, reliable, time saving, and applicable to treating a large number of the clinical samples.

Human influenza infections display a strongly seasonal pattern. However, whether H7N9 and H5N1 infections correlate with climate factors has not been examined. Here, we analyzed 350 cases of H7N9 infection and 47 cases of H5N1 infection. The spatial characteristics of these cases revealed that H5N1 infections mainly occurred in the South, Middle, and Northwest of China, while the occurrence of H7N9 was concentrated in coastal areas of East and South of China. Aside from spatial-temporal characteristics, the most adaptive meteorological conditions for the occurrence of human infections by these two viral subtypes were different. We found that H7N9 infections correlate with climate factors, especially temperature (TEM) and relative humidity (RHU), while H5N1 infections correlate with TEM and atmospheric pressure (PRS). Hence, we propose a risky window (TEM 4-14 °C and RHU 65-95%) for H7N9 infection and (TEM 2-22 °C and PRS 980-1025 kPa) for H5N1 infection. Our results represent the first step in determining the effects of climate factors on two different virus infections in China and provide warning guidelines for the future when provinces fall into the risky windows. These findings revealed integrated predictive meteorological factors rooted in statistic data that enable the establishment of preventive actions and precautionary measures against future outbreaks.

Background: A new strain of human H1N1 influenza A viruses was broken out in the April 2009 and caused worldwide pandemic emergency. The present study is trying to estimate a temporal reassortment history of 2009 H1N1 viruses by phylogenetic analysis based on a total 394 sequences of H1N1viruses isolated from swine, human and avian., Results: Phylogenetic trees of eight gene segments showed that viruses sampled from human formed a well-supported clade, whereas swine and avian lineages were intermixed together. A new divergence swine sublineage containing gene segments of 2009 H1N1 viruses was characterized, which were closely related with swine viruses collected from USA and South Korea during 2004 to 2007 in six segments (PB2, PB1, PA, HA, NP and NS), and to swine viruses isolated from Thailand during 2004 to 2005 in NA and M. Substitution rates were varied drastically among eight segments and the average substitution rate was generally higher in 2009 H1N1 than in swine and human viruses (F2,23 = 5.972, P < 0.01). Similarly, higher dN/dS substitution ratios were identified in 2009 H1N1 than in swine and human viruses except M2 gene (F2, 25 = 3.779, P < 0.05). The ages of 2009 H1N1 viruses were estimated around 0.1 to 0.5 year, while their common ancestors with closest related swine viruses existed between 9.3 and 17.37 years ago., Conclusion: Our results implied that at least four reassortments or transmissions probably occurred before 2009 H1N1 viruses. Initial reassortment arose in 1976 and avian-like Eurasian swine viruses emerged. The second transmission happened in Asia and North America between 1988 and 1992, and mostly influenced six segments (PB2, PB1, PA, HA, NP and NS). The third reassortment occurred between North American swine and avian viruses during 1998 to 2000, which involved PB2 and PA segments. Recent reassortments occurred among avian-to-swine reassortant, Eurasian and classical swine viruses during 2004 to 2005. South Korea, Thailand and USA, were identified as locations where reassortments most likely happened. The co-circulation of multiple swine sublineages and special lifestyle in Asia might have facilitated mixing of diverse influenza viruses, leading to generate a novel virus strain.

The schizothoracine Gymnocypris chilianensis is restricted to the Shiyang, Ruoshui and Shule Rivers, listed from east to west, along the northeast edge of the Tibetan Plateau. This distribution provides a valuable system to test hypotheses about postglacial colonization. We used mitochondrial DNA sequence data (a control region and the cytochrome b gene; 1894 bp) to assess the phylogeographic structure of this species based on 278 specimens sampled from throughout the species' entire geographical range. We found three lineages corresponding geographically to the three rivers, suggesting three independent glacial differentiation centers within the northeast edge of the Tibetan Plateau. The phylogenetic analysis suggested that the Shiyang River population forms a lineage that separated from the other populations of G. chilianensis at the basal phylogenetic split within this species. The molecular data further demonstrated a clear pattern of decreasing genetic diversity from the eastern Shiyang River towards the central Ruoshui River and western Shule River lineages, a pattern consistent with sequential western colonization. We therefore propose a phylogeographic scenario for G. chilianensis of a gradual westerly expansion from the Shiyang River population along the northeast edge of the Tibetan Plateau, with subsequent allopatric evolution at approximately 0.37 and 0.05 million years ago (Ma), through at least two glacial maxima. Together with the genetic evidence reported in other species, our findings suggest that this common biogeographic pattern emphasizes the importance of the northeastern edge region of the Tibetan Plateau as a hotspot of genetic diversity for some taxa., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Epithelial-derived thymic stromal lymphopoietin (TSLP) is an IL-7-like cytokine that is mainly produced by epithelial cells. It has been shown to play a key role in the development of Th2-type allergic inflammation. Commercial antibodies available against TSLP can only be used in Western blot assay, which further limits investigation of its function. Here we efficiently generated a panel of anti-mouse TSLP monoclonal antibodies in rats with DNA priming-protein boosting strategy. Overall, four MAb strains (4B12, 4E11, 5D7, and 6H10) were obtained and their characterizations were identified. The MAbs can specifically bind to TSLP according to the ELISA and FACS assays. It was found that they recognized distinct epitopes. They are useful in detecting TSLP expression in the tissue by immunoblotting and in the cytoplasm by intracellular staining assay. Thus, these antibodies will be valuable tools for studying TSLP biological functions.

Intron retention in histamine H(3) receptors has been found in many mammals, including rats and mice. The short transcript isoforms that exclude alternatively spliced introns are readily detected in very low abundance in rats and are undetectable in mice using the regular PCR approach. The detailed mechanism for the special alternative splicing remains poorly understood. The aim of this work was to investigate the effects of essential splice signals on intron retention and to identify sequence elements that determine the differences in splicing between rats and mice. We have constructed a minigene-splicing system to mimic natural alternative receptor splicing and analyzed the regulatory elements in HEK293 cells. Mutating suboptimal 5' and 3' splice sites toward the consensus sequences can enhance the splicing of corresponding alternative introns. The effect is much more significant for the 3' splice site of the longer intron than for the other two splice sites; it is also more significant in rats than in mice. The splicing differences between rats and mice are primarily determined by the six discrepant nucleotides within the alternative introns, which promote or suppress the intron splicing in different ways and cooperate to make splicing of the two introns more efficient in rats. From these results we conclude that (1) the weakness of the splice sites is an important determinant for very low efficiency during intron splicing and, (2) multiple sequence elements determine the splicing differences between rats and mice. The results provide insight into special alternative splicing regulation in H(3) receptors.

In the alternative splicing, intron retention, of histamine H(3) receptors in rats and mice, the short transcript isoforms that are excised alternatively spliced introns are easily detected in a very low level in rats and are undetectable in mice using the regular PCR protocol. The retained introns have common 5' splice site and different 3' splice sites. The detailed mechanism for the special alternative splicing remains largely unclear. In this study, we developed a minigene splicing system to recapitulate natural alternative splicing of the receptors and investigated the effects of 5' and 3' splice sites on intron retention in HeLa cells. Mutating weak 5' and 3' splice sites of the alternatively spliced introns toward the canonical consensus sequences promoted the splicing of the corresponding introns in rat and mouse minigenes. The effect of splice site strength was context-dependent and much more significant for the 3' splice site of the longer alternative intron than for the 3' splice site of the shorter alternative intron and the common 5' splice sites; it was also more significant in the rat minigene than in the mouse minigene. Mutating the 3' splice site of the longer alternative intron resulted in almost complete splicing of the intron and made the corresponding isoform to become the nearly exclusive transcript in the rat minigene.

Biomedical research in human beings is largely restricted to in vitro studies that lack complexity of a living organism. To overcome this limitation, humanized mouse models are developed based on immunodeficient characteristics of severe combined immunodeficiency (SCID) or recombination activating gene (Rag)(null) mice, which can accept xenografts. Peripheral constitution of human immunity in SCID or Rag(null) mice has been achieved by transplantation of mature human immune cells, foetal human thymus, bone marrow, liver tissues, lymph nodes or a combination of these, although efficiency needs to be improved. These mouse models with constituted human immunity (defined as humanized mice in the present text) have been widely used to investigate the basic principles of human immunobiology as well as complex pathomechanisms and potential therapies of human diseases. Here, elements of an ideal humanized mouse model are highlighted including genetic and non-genetic modification of recipient mice, transplantation strategies and proposals to improve engraftments. The applications of the humanized mice to study the development and response of human immune cells, human autoimmune diseases, virus infections, transplantation biology and tumour biology are reviewed as well.

Bone morphogenetic protein 2 (BMP2) has great potential in therapeutic applications. We are working on generating transgenic plants as a bioreactor to produce BMP2. We have studied the effects of codon optimization on the expression of human BMP2 (hBMP2) in tobacco plants. Three modified hBMP2 genes were transformed into tobacco under the control of either cauliflower mosaic virus 35S (CaMV35S) promoter or double-CaMV35S promoter plus alfalfa mosaic virus (AMV) enhancer. The fused beta-glucuronidase (GUS) reporter gene was used to facilitate the assay of protein expression. The results indicated that codon optimization could increase the protein expression level obviously under CaMV35S promoter. However, under relatively stronger initiation condition (double-CaMV35S promoter plus AMV enhancer), only the gene with the lowest degree of codon optimization could increase the protein expression level. Our findings suggest that the action of codon optimization may be influenced by the factors of promoter strength and A+T content in tobacco plants.

Cells are the functional elements in tissue engineering and regenerative medicine. A large number of cells are usually needed for these purposes. However, there are numbers of limitations for in vitro cell proliferation. Nanog is an important self-renewal determinant in embryonic stem cells. However, it remains unknown whether Nanog will influence the cell cycle and cell proliferation of mature cells. In this study, we expressed Nanog in NIH3T3 cells and showed that expression of Nanog in NIH3T3 promoted cells to enter into S phase and enhanced cell proliferation. This suggests that Nanog gene might function in a similar fashion in mature cells as in ES cells. In addition, it may provide an approach for in vitro cell expansion.

The conserved sequences of the mouse histamine H3 receptor at the potential alternative splice junctions suggest that the splice isoforms found in guinea pig, rat, human, and hamster may also be present in the mouse. However, the trace amount isoforms are hard to be detected by the regular PCR approach. In this paper, we report a method in which the unspliced long isoform is cut by restriction endonuclease so that the short isoforms can be amplified to detectable levels to confirm the existence of the splice isoforms of H3 receptor mRNA in the mouse. This method is applicable to the detection of trace amounts of splice isoforms that coexist with the long, more abundant isoforms.