Chromatin accessibility and transcriptional landscape in PK-15 cells during early exposure to Aflatoxin B1.

Aflatoxin B1 (AFB1) not only causes significant losses in livestock production but also poses a serious threat to human health. It is the most carcinogenic among known chemicals. Pigs are more susceptible to AFB1 and experience a higher incidence. However, the molecular mechanism of the toxic effect of AFB1 remains unclear. In this study, we used assay for transposase-accessible chromatin using sequencing (ATAC-seq) and RNA-seq to uncover chromatin accessibility and gene expression dynamics in PK-15 cells during early exposure to AFB1. We observed that the toxic effects of AFB1 involve signaling pathways such as p53, PI3K-AKT, Hippo, MAPK, TLRs, apoptosis, autophagy, and cancer pathways. Basic leucine zipper (bZIP) transcription factors (TFs), including AP-1, Fos, JunB, and Fra2, play a crucial role in regulating the biological processes involved in AFB1 challenge. Several new TFs, such as BORIS, HNF1b, Atf1, and KNRNPH2, represent potential targets for the toxic mechanism of AFB1. In addition, it is crucial to focus on the concentration of intracellular zinc ions. These findings will contribute to a better understanding of the mechanisms underlying AFB1-induced nephrotoxicity and offer new molecular targets.

HK2 and LDHA upregulation mediate hexavalent chromium-induced carcinogenesis, cancer development and prognosis through miR-218 inhibition.

Hexavalent chromium [Cr(VI)] is one of the most common environmental contaminants due to its tremendous industrial applications, but its effects and mechanism remain to be investigated. Our previous studies showed that Cr(VI) exposure caused malignant transformation and tumorigenesis. This study showed that glycolytic proteins HK2 and LDHA levels were statistically significant changed in blood samples of Cr(VI)-exposed workers and in Cr-T cells compared to the control subjects and parental cells. HK2 and LDHA knockdown inhibited cell proliferation and angiogenesis, and higher HK2 and LDHA expression levels are associated with advanced stages and poor prognosis of lung cancer. We found that miR-218 levels were significantly decreased and miR-218 directly targeted HK2 and LDHA for inhibiting their expression. Overexpression of miR-218 inhibited glucose consumption and lactate production in Cr-T cells. Further study found that miR-218 inhibited tumor growth and angiogenesis by decreasing HK2 and LDHA expression in vivo. MiR-218 levels were negatively correlated with HK2 and LDHA expression levels and cancer development in human lung and other cancers. These results demonstrated that miR-218/HK2/LDHA pathway is vital for regulating Cr(VI)-induced carcinogenesis and human cancer development.

High performance laser-driven flyers based on a refractory metamaterial perfect absorber.

Laser-driven flyers (LDFs), which can drive metal particles to ultra-high speeds by feeding high-power laser, have been widely used in many fields, such as ignition, space debris simulation, and dynamic high-pressure physics. However, the low energy-utilization efficiency of the ablating layer hinders the development of LDF devices towards low power consumption and miniaturization. Herein, we design and experimentally demonstrate a high-performance LDF based on the refractory metamaterial perfect absorber (RMPA). The RMPA consists by a layer of TiN nano-triangular array, a dielectric layer and a layer of TiN thin film, and is realized by combing the vacuum electron beam deposition and colloid-sphere self-assembled techniques. RMPA can greatly improve the absorptivity of the ablating layer to about 95%, which is comparable to the metal absorbers, but obviously larger than that of the normal Al foil (∼10%). This high-performance RMPA brings a maximum electron temperature of ∼7500 K at ∼0.5 µs and a maximum electron density of ∼1.04 × 1016 cm-3 at ∼1 µs, which are higher than that the LDFs based on normal Al foil and metal absorbers due to the robust structure of RMPA under high-temperature. The final speed of the RMPA-improved LDFs reaches to about 1920 m/s measured by the photonic Doppler velocimetry system, which is about 1.32 times larger than the Ag and Au absorber-improved LDFs, and about 1.74times larger than the normal Al foil LDFs under the same condition. This highest speed unambiguously brings a deepest hole on the Teflon slab surface during the impact experiments. The electromagnetic properties of RMPA, transient speed and accelerated speed, transient electron temperature and density have been systematically investigated in this work.

Prevalence and multilocus analysis of Giardia duodenalis in racehorses in China.

Giardia duodenalis is a zoonotic intestinal parasite infecting humans and mammals worldwide. In this study, we evaluated the prevalence of G. duodenalis in racehorses in China and genetically characterized it. In total, 621 fecal samples were collected from racehorses at 17 equestrian clubs in 15 cities in China. Forty-eight (7.7%) animals from 11 equestrian clubs were positive for G. duodenalis of assemblages A (n = 10), B (n = 36), and E (n = 2), based on the small subunit ribosomal RNA (SSU rRNA) gene. Statistically significant differences in the prevalence of this parasite were detected among the different equestrian clubs (χ2 = 49.55, df = 16, p < 0.01), whereas no significant differences were detected according to age (χ2 = 0.64, df = 1, p > 0.05) or sex (χ2 = 1.41, df = 2, p > 0.05). The G. duodenalis-positive samples were further subtyped based on three other genes, which identified 5, 4, and 4 genotypes at the triose phosphate isomerase (tpi), glutamate dehydrogenase (gdh), and ß-giardin (bg) loci, respectively. Subassemblage BIV was the predominant genotype. A phylogenetic analysis of the concatenated sequences of subassemblage BIV showed that the multilocus genotypes from the horses were genetically different from those of humans and nonhuman primates, indicating the evolution of host separation in G. duodenalis subassemblage BIV. Our study extends our understanding of the transmission of G. duodenalis between animals and humans.

Perceptions of the motivational climate, basic psychological needs, and life skills development in Chinese physical education students.

Introduction: Life skills can have a positive impact on young people's mental health, academic performance, and overall well-being. Physical education (PE) is viewed as a promising setting for developing students' life skills, but less is known about this in non-English speaking countries such as China. Based on the integration of Self-Determination Theory (SDT) and Achievement Goal Theory (AGT), we aimed to examine the relationships between students' perceptions of the teacher-initiated motivational climate (mastery- or performance-oriented) and their life skills development in PE, as well as the mediating role of their basic psychological needs (BPNs) (satisfaction or frustration). Methods: We employed a cross-sectional survey. Chinese students (N = 533, Age range = 13-18 years) completed measures assessing these variables. We fulfilled correlational and mediational analyses. Results: These findings showed that mastery climate was positively associated with needs satisfaction (r = 0.66) and eight life skills (r range = 0.44-0.61), whereas negatively associated with needs frustration (r = -0.49). Performance climate was positively related to needs frustration (r = 0.52), but negatively related to needs satisfaction (r = -0.38) and eight life skills (r range = -0.28 - -0.15). Needs satisfaction was positively (r range = 0.44-0.65), while needs frustration was negatively (r range = -0.50 - -0.34) linked with eight life skills. Furthermore, needs satisfaction positively mediated the effect of mastery climate on life skills development, but it is not found that needs frustration mediated the effect of performance climate on life skills besides goal setting, social skills, and time management. Conclusion: In conclusion, our study extended the previous literature on life skills in PE, and highlighted the roles of motivational climate and BPNs on students' life skills development. In practice, PE teachers should be encouraged to create a mastery climate as well as avoid a performance climate, to foster students' BPNs satisfaction, which in turn, promote their life skills development.

Biphasic function of GSK3ß in gefitinib­resistant NSCLC with or without EGFR mutations.

Epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs), such as gefitinib, are effective in the treatment of non-small cell lung cancer (NSCLC) harboring EGFR mutations. However, the mechanism underlying acquired resistance to EGFR-TKIs remains largely unknown. Therefore, the present study generated gefitinib-resistant PC-9 (PC-9G) cells, which were revealed to be more resistant to gefitinib-induced reductions in proliferation, migration and invasion, and increases in apoptosis, and had no detectable EGFR mutations compared with the control PC-9 cell line. In addition, the present study performed genome-wide transcriptomic analysis of differentially expressed genes between PC-9 and PC-9G cell lines. Cell proliferation, colony formation, invasion, migration and flow cytometry analyses were also performed. The genome-wide transcriptomic analysis revealed that glycogen synthase kinase 3ß (GSK3ß) was downregulated in PC-9G cells compared with that in PC-9 cells. Furthermore, GSK3ß overexpression increased the proliferation, migration and invasion of PC-9 and H1975 gefitinib-resistant cells. Conversely, overexpression of GSK3ß suppressed the proliferation, migration and invasion of PC-9G cells. Furthermore, AKT inhibition reduced the proliferation, migration and invasion, and induced the apoptosis of PC-9, PC-9G and H1975 cells, the effects of which were reversed following AKT activation; notably, the tumor suppressor function of GSK3ß was inconsistent with the tumor promotor role of the AKT pathway in PC-9G cells without EGFR mutation. The present study may provide novel insights into the distinctive role of GSK3ß in gefitinib-resistant NSCLC with or without EGFR mutations, suggesting that a more detailed investigation on GSK3ß as a therapeutic target for gefitinib-resistant NSCLC may be warranted.

Functional Mutations in the microRNA-155 Promoter Modulate its Transcription Efficiency and Expression.

Limited research has been conducted on porcine miR-155 promoters, and previous study from our group have identified two haplotypes (TT and CC) in different pig breeds, each associated with five fully linked mutation sites within or near the miR-155 gene (Li et al. Dev Comp Immunol 39(1):110-116, 2013). In this study, the promoter region of porcine miR-155 was screened, and two important transcription factors, Foxp3 and RelA, were identified. The binding ability of Foxp3 protein was found to be affected by the first mutation site (A/C) using EMSA analysis. In vitro experiments revealed that the expression level of miR-155 was significantly higher in the C haplotype compared to the T haplotype. Additionally, northern blotting assays indicated that both the first mutation site (A/C) and the fourth mutation site (G/T) had a significant impact on miR-155 expression levels. These findings provide further insights into the transcriptional regulation of porcine miR-155 and identify crucial mutation sites that influence miR-155 expression. This knowledge can serve as a basis for identifying potential molecular markers associated with disease resistance in swine.

Development and Psychometric Evaluation of the Chinese Version of the Life Skills Scale for Physical Education.

Research on life skills in physical education (PE) has gained great attention in recent years. However, there is a need to translate life skills measures for PE into other languages. This research adapted the Life Skills Scale for PE (LSSPE) into Chinese and provided evidence for its validity and reliability. In Study 1, the scale was cross-culturally adapted through translation and back-translation, expert feedback, pilot testing, and scale refinement to provide evidence for the content validity of the scale. Study 2 provided evidence for the factorial validity, internal consistency reliability, and test-retest reliability of the scale by testing it with 583 students. Study 3, with 390 students, provided evidence for the nomological validity of the measure, with results showing perceived teacher autonomy support and students' basic need satisfaction were positively associated with life skills development in PE, and that controlling teaching and basic need frustration were negatively associated with life skills development. In conclusion, the results illustrate that the LSSPE can be used to evaluate Chinese-speaking students' life skills development in PE.

Research progress in toxicological effects and mechanism of aflatoxin B1 toxin.

Fungal contamination of animal feed can severely affect the health of farm animals, and result in considerable economic losses. Certain filamentous fungi or molds produce toxic secondary metabolites known as mycotoxins, of which aflatoxins (AFTs) are considered the most critical dietary risk factor for both humans and animals. AFTs are ubiquitous in the environment, soil, and food crops, and aflatoxin B1(AFB1) has been identified by the World Health Organization (WHO) as one of the most potent natural group 1A carcinogen. We reviewed the literature on the toxic effects of AFB1 in humans and animals along with its toxicokinetic properties. The damage induced by AFB1 in cells and tissues is mainly achieved through cell cycle arrest and inhibition of cell proliferation, and the induction of apoptosis, oxidative stress, endoplasmic reticulum (ER) stress and autophagy. In addition, numerous coding genes and non-coding RNAs have been identified that regulate AFB1 toxicity. This review is a summary of the current research on the complexity of AFB1 toxicity, and provides insights into the molecular mechanisms as well as the phenotypic characteristics.

Signifying the Relationship between Fear of COVID-19, Psychological Concerns, Financial Concerns and Healthcare Employees Job Performance: A Mediated Model.

The coronavirus pandemic (COVID-19) has undoubtedly created immense health problems in the global healthcare sector. Apart from its impact on physical health, it has devastatingly affected the psychological well-being of individuals. Based on Affective Events Theory (AET), the current study aims to contemplate the relationship between Fear of COVID-19 (CVF), psychological concerns (PC), and financial concerns (FC) while considering the impact on the healthcare employees' job performance (JP). Moreover, this study investigates the mediating role of job anxiety (ANX), stress (ST), and depression (DEP). The data were collected through an online structured questionnaire (Google Forms) from 489 employees working in the healthcare centers of Pakistan. The structured equation modeling (partial least square) approach is adopted for data analysis. The study results showed that Fear of COVID-19, psychological and financial concerns positively and significantly affect healthcare workers' job performance. Depression, anxiety, and stress mediated the relationship between Fear of COVID-19, psychological concerns, financial concerns, and job performance. The study theoretically and practically contributes to the existing literature on psychological and mental health by providing a better understanding of the individual variables that affect employees' job performance.

Corrigendum: Regulation of microrna-497-targeting AKT2 influences tumor growth and chemoresistance to cisplatin in lung cancer.

[This corrects the article DOI: 10.3389/fcell.2020.00840.].

Dominance of the zoonotic pathogen Cryptosporidium meleagridis in broiler chickens in Guangdong, China, reveals evidence of cross-transmission.

BACKGROUND: Cryptosporidium is one of the most prevalent parasites infecting both birds and mammals. To examine the prevalence of Cryptosporidium species and evaluate the public health significance of domestic chickens in Guangdong Province, southern China, we analyzed 1001 fecal samples from 43 intensive broiler chicken farms across six distinct geographical regions. METHODS: Individual DNA samples were subjected to nested PCR-based amplification and sequencing of the small subunit of the nuclear ribosomal RNA gene (SSU rRNA). Analysis of the 60 kDa glycoprotein gene (gp60) was performed to characterize the subtypes of C. meleagridis. RESULTS: The overall prevalence of Cryptosporidium was 13.2% (95% CI 11.1-15.3) (24 of 43 farms), with C. meleagridis (7.8%), C. baileyi (4.8%) and mixed infections (0.6%). Using the gp60 gene, three subtype families, IIIb, IIIe and IIIg, were identified, including six subtypes: one novel (IIIgA25G3R1a) and five previously reported (IIIbA23G1R1c, IIIbA24G1R1, IIIbA21G1R1a, IIIeA17G2R1 and IIIeA26G2R1). Within these subtypes, five known subtypes were genetically identical to those identified in humans. CONCLUSIONS: This is the first report of C. meleagridis in chickens from Guangdong. The frequent occurrence of C. meleagridis in domestic chickens and the common C. meleagridis subtypes identified in both humans and chickens is of public health significance. Our study indicates that broiler chickens represent a potential zoonotic risk for the transmission of Cryptosporidium in this region.

MiRNA-30e downregulation increases cancer cell proliferation, invasion and tumor growth through targeting RPS6KB1.

Human esophagus carcinoma (EC) is one of the most common malignant tumors, especially in Africa and Asia including China. In EC initiation and progression, genetic and epigenetic aberrations have been reported to play a major role, but the underlying molecular mechanisms are largely unknown. In this study, the miR-30e levels were analyzed in human EC tissues and TCGA databases, and the results demonstrated that miR-30e expression in EC tissues was significantly decreased compared to adjacent normal tissues. To further investigate the role of miR-30e in cancer cells, we found that forced expression of miR-30e dramatically inhibited cell proliferation, invasion, tube formation, and colony formation of cancer cells. To determine the underlying mechanism of miR-30e, we found that RPS6KB1 was a direct target of miR-30e by binding to its 3'-UTR, which was verified by luciferase activity assay using reporters with wild-type miR-30e and its seed sequence mutant constructs and Western blotting assay. In vivo experiment showed that miR-30e overexpression significantly inhibited tumor growth and decreased RPS6KB1 expression in xenografts. In EC, high expression of RPS6KB1 in tumor tissues indicated poor prognosis of patients with less survival rate. High levels of RPS6KB1 and low levels of miR-30e closely correlated poor survival of patients with several other types of cancer. These findings show that miR-30e and its target RPS6KB1 are important in cancer development and clinical outcomes, and miR-30e/RPS6KB1 is a potential future therapeutic pathway for EC intervention.

Improving the Energy Conversion Efficiency of a Laser-Driven Flyer by an In Situ-Fabricated Nano-absorption Layer.

Three kinds of Al flyer plates with different nanostructured absorption layers were in situ prepared by a direct laser writing technology to improve the energy conversion efficiency in a laser-driven flyer assembly. Microstructures, light absorption, and flyer velocity in the acceleration chamber were investigated. The reflectance for the flyers at 1064-nm wavelength can be reduced from 81.3 to 9.8% by the nanostructured absorption layer. The terminal velocity of a 50-µm-thick Al flyer irradiated by a 60-mJ laser pulse is 831 m/s, while the velocity of the flyer with an in situ-fabricated nano-absorption layer reaches up to 1113 m/s at the same condition. Resultantly, the energy conversion efficiency of the flyer with a nanostructure absorption layer can reach as high as 1.99 times that of the Al flyer. Therefore, the nanostructured absorption layer in situ prepared on the surface of a flyer provides a new method to significantly improve the energy conversion efficiency of a laser-driven flyer.

Regulation of MicroRNA-497-Targeting AKT2 Influences Tumor Growth and Chemoresistance to Cisplatin in Lung Cancer.

BACKGROUND: MicroRNA-497 (miR-497) has been implicated in several cancers. Increasing studies demonstrate the role of AKT2 in cancers as an oncogene which is closely associated with tumor aggressiveness by enhancing cancer cell survival, migration and invasion However, miR-497/AKT2 axis in non-small cell lung cancer (NSCLC) remains unclear. METHODS: Quantitative real-time PCR (qRT-PCR) was used to quantify the expression of miR-497 and its target gene. The function of miR-497 in lung cancer was investigated through in vitro and in vivo assays (cell proliferation assay, cell migration assay, colony formation assay, flow cytometry assay, immunoblotting and tumorigenesis assay). Luciferase reporter assay was conducted to confirm the target gene of miR-497. RESULTS: In this study, we found that miR-497 was significantly downregulated in tumor tissues and blood samples of lung cancer patients. To understand the potential mechanism of miR-497 in inhibiting tumor growth, we showed that miR-497 blocked the activation of AKT2 and regulated cell proliferation, cell migration, colony formation and increases chemosensitivity of H1299 cells to cisplatin by inhibiting AKT2. MiR-497 also inhibited tumor growth and suppressed expression of AKT2 at the protein and mRNA levels in mouse xenograft tumors. CONCLUSION: Taken together, our findings indicated that miR-497 suppresses the tumor growth by targeting AKT2, and the miR-497/AKT2 axis is a potential therapeutic target for NSCLC intervention.

Apigenin Inhibits IL-6 Transcription and Suppresses Esophageal Carcinogenesis.

Esophagus cancer is the seventh cause of cancer-related deaths globally. In this study, we analyzed interleukin 6 (IL-6) gene expression in human esophagus cancer patients and showed that IL-6 mRNA levels are significantly higher in tumor tissues and negatively correlated with overall survival, suggesting that IL-6 is a potential therapeutic target for esophagus cancer. We further demonstrated that apigenin, a nature flavone product of green plants, inhibited IL-6 transcription and gene expression in human esophagus cancer Eca-109 and Kyse-30 cells. Apigenin significantly and dose-dependently inhibited cell proliferation and promoted apoptosis while stimulating the cleaved PARP (poly ADP-ribose polymerase) (C-PARP) and caspase-8 expression. It suppressed VEGF (Vascular endothelial growth Factor) expression and tumor-induced angiogenesis. Pretreatment of cells with IL-6 could completely reverse apigenin-induced cellular changes. Finally, using a preclinical nude mice model subcutaneously xenografted with Eca-109 cells, we demonstrated the in vivo antitumor activity and mechanisms of apigenin. Taken together, this study revealed for the first time that apigenin is a new IL-6 transcription inhibitor and that inhibiting IL-6 transcription is one of the mechanisms by which apigenin exhibits its anticancer effects. The potential clinical applications of apigenin in treating esophagus cancer warrant further investigations.

MiR-199a Inhibits Tumor Growth and Attenuates Chemoresistance by Targeting K-RAS via AKT and ERK Signalings.

Glioma is the most malignant brain tumors in the world, the function and molecular mechanism of microRNA-199a (miR-199a) in glioma is not fully understood. Our research aims to investigate miR-199a/K-RAS axis in regulation of glioma tumor growth and chemoresistance. The function of miR-199a in glioma was investigated through in vitro and in vivo assays. We found that miR-199a in tumor tissues of glioma patients was significantly downregulated in this study. Kinase suppressor of ras 1 (K-RAS), was indicated as a direct target of miR-199a, as well as expression levels of K-RAS were inversely correlated with expression levels of miR-199a in human glioma specimens. Forced expression of miR-199a suppressed AKT and ERK activation, decreased HIF-1α and VEGF expression, inhibited cell proliferation and cell migration, forced expression of K-RAS restored the inhibitory effect of miR-199a on cell proliferation and cell migration. Moreover, miR-199a renders tumor cells more sensitive to temozolomide (TMZ) via targeting K-RAS. In vivo experiment validated that miR-199a functioned as a tumor suppressor, inhibited tumor growth by targeting K-RAS and suppressed activation of AKT, ERK and HIF-1α expression. Taken together, these findings indicated that miR-199a inhibits tumor growth and chemoresistance by regulating K-RAS, and the miR-199a/K-RAS axis is a potential therapeutic target for clinical intervention in glioma.

[Prokaryotic expression of N-terminal antigenic domain of duck plague virus gB protein and the establishment of putative indirect ELISA assay].

Based on the antigenic analysis of duck plague virus (DPV) gB protein, we designed a pair of primers to amplify the gene fragment encoding high antigenic domain of DPV N-terminal gB protein from the DPV genome. The cloned gene was digested with EcoR I and Hind III and then inserted into pET32a vector to obtain the recombinant pET-gB1 plasmid. The recombinant plasmid was transformed into E. coli BL21, and expressed in very high level after induced with IPTG. The expressed product was analyzed by SDS- PAGE and Western blotting. The result indicated that the fusion protein (pET-gB1) existed as inclusion body, which was about 42.4kDa and showed specific immunoreactivity with anti-DPV sera. The recombinant gB1 protein was purified with His-Bind resin protein purification procedure. Then an indirect ELISA was established to detect antibody against DPV with the purified gB1 protein as the coating antigen. The result showed that the optimal concentration of coated antigen was 6.5 microg/mL and the optimal dilution of serum was 1 : 80. The positive criterion of this ELISA assay was OD(the tested serum) > 0.4 and OD(the tested serum)/OD(the negative serum) > 2.0. The ELISA was done on 700 sera that were preserved in Shandong, Jiangsu Provinces, and were detected by igB1-ELISA and iDPV-ELISA with duck plague virus as the coating antigen respectively. The agreement ratio between the two methods was 95.6%.

The immunomodulatory peptide bursopentin (BP5) enhances proliferation and induces sIgM expression in DT40 cells.

BACKGROUND: In the recent past, many studies have been focused on extracts of BF and multiple biologically active factors and their effects on humoral immune system in chickens and birds. However, the mechanism of those immunomodulatory peptides on the B lineage cells proliferation and antibody production in chicken is fairly unknown. DT40 cell line, an avian leucosis virus-induced chicken pre-B cell line, expresses immunoglobulin M (IgM) isotype B cell reporter in the plasma membrane. There are many evidences suggesting that DT40 cells are best characterized as a bursal stem cell line. Because of the unique characteristics of DT40 cell line, it has been widely used to observe biological processes of pre-B lymphocyte cell within living cells. METHODS: The chicken B cell line DT40 was cultured in Roswell Park Memorial Institute (RPMI) 1640 medium and cytotoxicity was studied. Also, effect of BP5 on cell proliferation and cell cycle distribution of DT40 cells was studied. Also, the effect of BP5 on sIgM mRNA expression was studied by using real-time PCR. OBJECTIVES: To investigat the effects of Bursopentin (Cys-Lys-Arg-Val-Tyr, BP5) on a chicken promyelocyte cell line DT40, assays of cell proliferation, cell cycle distribution, detection of surface immunoglobulin G (sIgM) mRNA expression and gene microarray analysis were performed. RESULTS: The results showed that BP5 displayed concentration-dependent effects on the proliferation, cell cycle, and sIgM mRNA expression in DT40 cells. And the analysis of expression profiles identified a signature set of 3022 genes (1254 up regulated genes, 1762 down regulated genes), which clearly discriminated the BP5-treated DT40 cells from control with high certainty (P≤0.02). The results of microarray analysis were confirmed by quantitative reverse transcription-polymerase chain reaction for 12 of the differentially expressed genes. CONCLUSION: Theses findings showed the immuno-activity effect of BP5 on B lymphocyte and indicated that BP5 treatment regulated eight signaling pathways, in which Toll-like signaling pathway was the most significant enrichment pathway.

Immunomodulatory roles and functional analysis of pre-B lymphocyte DT40 cells with the bursal-derived BSP-II treatment.

The bursa of Fabricius, the acknowledged central humoral immune organ, is vital to B cell differentiation. However, the regulatory function of the bursal-derived peptide on avian B cell proliferation has not been reported. BSP-II is a recently reported bursal-derived bioactive peptide. In this paper, 75 days-old chicks were twice subcutaneously immunized with BSP-II and inactivated avian influenza virus (AIV, H(9)N(2) strain). It was proved that BSP-II induced a strongly AIV-specific HI antibody production in the immunized chicks. Also, BSP-II could enhance avian pre-B lymphocyte DT40 cell viability. To investigate the global patterns of gene expression in DT40 cells after BSP-II treatment, gene microarray was carried out. It was identified that the differentially expressed genes were involved in various pathways, of which six pathways were associated with signaling transductions, including ErbB signaling, MAPK signaling, Toll-like receptor signaling, Notch signaling, mTOR signaling, and Wnt signaling. Finally, RT-qPCR was used to confirm the microarray expression data. These results indicated the molecular basis of pre-B lymphocyte viability with BSP-II treatment, which provided a potential mechanism of the bursa of Fabricius on pre-B lymphocyte viability, differentiation, and development. These results are valid for the mechanism of the bursa of Fabricius on B lymphocytes development.