Transcript variants and expression profiles analysis of Mitf gene in minipigs

Object： To identify transcript variants and expression patterns of porcine Mitf. Materials and methods： A pairwise BLAST search at NCBI database was performed to deduce the structure of porcine Mitf gene. Subsequently, 5＇ RACE and fluorescent quantitative RT-PCR were used to analyze the expression pattern of porcine Mitf in different tissues. Results： Four transcript variants of porcine Mitf, MITF-A, MITF-H, MITF-M and MITF-SUS were identified, all sharing high homology with those in humans, except Mitf-SUS.Conclusion： The sequence of porcine Mitf appear highly homologous to human MITF. However, only 4 transcript variants of porcine Mitf were identified in these minipigs, less than the 9 transcript variants in human MITF.

Proliferating cell nuclear antigen clamp associated factor,a potential proto-oncogene with increased expression in malignant gastrointestinal tumors

Gastrointestinal(GI)cancers,including malignancies in the gastrointestinal tract and accessory organs of digestion,represent the leading cause of death worldwide due to the poor prognosis of most GI cancers.An investigation into the potential molecular targets of prediction,diagnosis,prognosis,and therapy in GI cancers is urgently required.Proliferating cell nuclear antigen(PCNA)clamp associated factor(PCLAF),which plays an essential role in cell proliferation,apoptosis,and cell cycle regulation by binding to PCNA,is a potential molecular target of GI cancers as it contributes to a series of malignant properties,including tumorigenesis,epithelial-mesenchymal transition,migration,and invasion.Furthermore,PCLAF is an underlying plasma prediction target in colorectal cancer and liver cancer.In addition to GI cancers,PCLAF is also involved in other types of cancers and autoimmune diseases.Several pivotal pathways,including the Rb/E2F pathway,NF-κB pathway,and p53-p21 cascade,are implicated in PCLAF-mediated diseases.PCLAF also contributes to some diseases through dysregulation of the p53 pathway,WNT signal pathway,MEK/ERK pathway,and PI3K/AKT/mTOR signal cascade.This review mainly describes in detail the role of PCLAF in physiological status and GI cancers.The signaling pathways involved in PCLAF are also summarized.Suppression of the interaction of PCLAF/PCNA or the expression of PCLAF might be potential biological therapeutic strategies for GI cancers.

Identification of Toll-like Receptor (TLR) 4 Gene in Wild Boar

Toll-like receptor （TLR） 4 plays an important role in the innate immune system and has been involved in resistance/ susceptibility to a number of diseases as revealed by studies in human and other domestic animals. Wild boar survives in natural environment without artificial interference and may be different from domestic pig in innate immune system. Here, the complete coding sequence of TLR4 and TLR4A was cloned in wild boar, and two other alternative splicing variants, TLR4B and TLR4C, were obtained. Compared to the counterpart from domestic pig （GcnBank No. AJ628065）, there were five SNPs, c.510T〉C, c.960A〉G, c.962A〉G, c.1605T〉G and c.1824A〉G, in the coding sequence of wild boar TLR4A gene. TLR4 gene was expressed in all the tissues from wild boar studied with the most abundance in spleen tissue, and mRNA level was significantly lower in spleen from wild boar than that from Min pig. The allele distribution was significantly different at polymorphic loci c.962G〉A and c.1027C〉A （p〈0.01） between wild boar and Min pig. The results would contribute to understand the innate immune system in wild boar.

Expression of PYCARD Gene Transcript Variant mRNA in Peripheral Blood Mononuclear Cells of Primary Gout Patients with Different Chinese Medicine Syndromes

Objective： To study the expression level and role of apoptosis-associated speck-like protein containing a caspase recruitment domain （PYCARD） gene transcript variant mRNA in peripheral blood mononuclear cells （PBMCs） of primary gout （PG） patients with different Chinese medicine （CM） syndromes. Methods： The expressions of PYCARD gene transcript variant mRNA and interleukin-1β （IL-1β） mRNA in PBMCs were investigated in 96 PG patients with acute phase （APPG, 44 cases） and non-acute phase （NAPPG, 52 cases） and 30 healthy controls （HCs） by reverse transcription-polymerase chain reaction （PCR） and/or real-time quantitative PCR. PYCARD and nuclear factor-κB （p50） [NF-κB （p50）] protein was detected by Western blot in PBMCs respectively. IL-1β, IL-4 and IL-10 protein levels in plasma of HCs and PG patients were measured by enzyme-linked immuno sorbent assay. Results： The main CM syndromes in APPG patients were obstruction of dampness and heat syndrome （ODHS, 36.36%） and intermingled phlegm-blood stasis syndrome （IPBSS, 27.27%）, while in NAPPG patients were Pi （Spleen）-deficiency induced dampness syndrome （PDIDS, 40.38%） and qi-blood deficiency syndrome （QBDS, 26.92%）. It showed statistical significances of the expressions of PYCARD gene and its transcript variant mRNA, the protein of PYCARD and NF-κB （p50） and the plasma IL-1β, IL-4 and IL-10 in APPG, NAPPG, ODHS, IPBSS, PDIDS and QBDS groups, compared with the HC group respectively （P〈0.05 or P〈0.01）. There were also significant differences of mRNA expressions of PYCARD-1 and PYCARD-2 as well as protein expressions of IL-1β, IL-4 and IL-10 among the 4 CM syndromes groups （P〈0.05 or P〈0.01）. Correlation analysis showed positive correlation between the mRNA expressions of PYCARD-1 gene transcript variant and IL-1β in APPG patients （r=0.3088, P=0.0183）. Conclusion： PYCARD gene and its transcript variant may play a critical and regulative role in the inflammatory response of PG patients with different phases and CM syndromes.

Analysis of Allele Specific Expression——A Survey

Allele specific expression is essential for cellular programming and development and the diversity of cellular phenotypes. Traditional analysis methods utilize RNA and depend on single nucleotide polymorphisms,thus to suffer from limited amount of materials for analysis. The rapid development of next-generation sequencing technologies provides more comprehensive and powerful approaches to analyze the genomic, epigenetic, and transcriptomic data, and further to detect and measure allele specific expressions. It will potentially enhance the understanding of the allele specific expressions, their complexities, and the effect on biological processes. In this paper, we extensively review the state-of-art enabling technologies and tools to analyze, detect, and measure allele specific expressions, compare their features, and point out the future trend of the methods.