Generation of Chimeric RNAs by cis-splicing of adjacent genes (cis-SAGe) in mammals.

Chimeric RNA molecules, possessing exons from two or more independent genes, are traditionally believed to be produced by chromosome rearrangement. However, recent studies revealed that cis-splicing of adjacent genes (cis- SAGe) is one of the major mechanisms underlying the formation of chimeric RNAs. cis-SAGe refers to intergenic splicing of directly adjacent genes with the same transcriptional orientation, resulting in read-through transcripts, termed chimeric RNAs, which contain sequences from two or more parental genes. cis-SAGe was first identified in tumor cells, since then its potential in carcinogenesis has attracted extensive attention. More and more scientists are focusing on it. With the development of research, cis-SAGe was found to be ubiquitous in various normal tissues, and might make a crucial contribution to the formation of novel genes in the evolution of genomes. In this review, we summarize the splicing pattern, expression characteristics, possible mechanisms, and significance of cis-SAGe in mammals. This review will be helpful for general understanding of the current status and development tendency of cis-SAGe.

Sequence and expression regulation of the BCL2L2 gene in pigs.

B cell lymphoma-2-like 2 (BCL2L2), an important regulator of apoptosis, plays vital roles in several physiological processes, as revealed by studies in humans and mice. However, reports on pig BCL2L2 are few, and the encoding gene has not been identified experimentally. This study was designed to clone the porcine BCL2L2 gene and its alternative splicing (AS) transcripts using molecular biological techniques and to analyze the regulatory mechanisms underlying transcription and translation. The BCL2L2 cDNA (V1) was 807 bp in length and encoded a polypeptide of 193 aa containing four BCL-2 homology domains. A total of nine AS transcripts were obtained, among which V2 and V3 differed from V1 in the 5' untranslated region (UTR). The core promoter was mapped to a range of -1102 to -759 bp (the first nucleotide of the start codon was designated as +1). There were several functional cis-elements, including one SP1 and two C/EBPα binding sites at around -759 bp. AS in the 5' UTR is involved in the regulation of gene expression, as revealed by dual-luciferase reporter and western blot analysis, and the secondary structure of the 5' UTRs may be the reason for the differential expression of V1-3. At the same time, an upstream open reading frame (ORF) existed in each of the three 5' UTRs, was found to repress the expression of the main ORF. Additionally, the roles of porcine BCL2L2 in cell proliferation and apoptosis were preliminarily analyzed. The results will contribute to further characterizing the role of BCL2L2.

Diagnostic value of elevated serum carbohydrate antigen 125 level in sarcoidosis.

BACKGROUND: Sarcoidosis is a multisystem disorder with unknown etiology, and it predominantly affects the lungs and intrathoracic lymph nodes. For patients with atypical clinical manifestations, the diagnosis of sarcoidosis is difficult and specific biomarkers may play an important role in assisting diagnosis. Previous research has demonstrated a correlation between sarcoidosis and increased carbohydrate antigen 125 (CA125), but remains a lack of large cohort studies to validate this observation. AIM: To compare serum CA125 levels in sarcoidosis patients and healthy controls, and explore whether CA125 can be used as a biomarker for the diagnosis of sarcoidosis. METHODS: In this study, the serum CA125 levels were measured by enzyme-linked immunosorbent assay in 108 consecutive sarcoidosis patients between June 2016 and December 2020 (31 males, 77 females; age at diagnosis 49.69 ± 9.10 years) and 112 healthy subjects. Data on the C-reactive protein, erythrocyte sedimentation rate, and angiotensin-converting enzyme were also collected. The association of serum CA125 levels with clinical, radiological, and respiratory functional characteristics was analyzed between patient groups with CA125 ≤ 35 U/mL or CA125 > 35 U/mL. RESULTS: We found that serum CA125 levels were higher in sarcoidosis patients compared to healthy controls (median: 44.78 vs 19.11 U/mL, P < 0.001). The area under the receiver operator characteristic was 0.9833 (95%CI: 0.9717-0.9949), and the best cutoff point was 32.33 U/mL. The elevated serum CA125 was notably associated with the percentage of predicted forced vital capacity (FVC%) and neutrophil-to-lymphocyte ratio (P = 0.043 and P = 0.038, respectively) in sarcoidosis patients. Multivariate analysis revealed that FVC% was a statistically notable predictor of elevated serum CA125 (P = 0.029). Also, our research revealed that compared to patients with Stage I of radiology classification, patients with Stage II and III showed a higher concentration of serum CA125 (46.16 ± 8.32 vs 41.00 ± 6.04 U/mL, P = 0.005, and 47.92 ± 10.10 vs 41.00 ± 6.04 U/mL, P = 0.002, respectively). CONCLUSION: Serum CA125 was highly increased in sarcoidosis patients and showed high efficiency for noninvasive diagnosis of the disease. In addition, abnormally elevated serum CA125 was correlated with pulmonary function and radiological Scadding's classification of sarcoidosis.

Isolation and characterization of porcine PILRB gene and its alternative splicing variants.

Paired immunoglobulin-like type 2 receptor (PILR)ß regulates inflammatory responses to pathogen infection, and therefore plays an important role in host disease resistance/susceptibility. However porcine PILRß remains poorly characterized. In this study, we obtained the cDNA (V1) of its encoding gene, PILRB, and three alternative splicing (AS) variants (V2-4). The complete coding sequence of V1 was 621 bp long encoding a polypeptide of 206 aa. Compared with V1, V2 and V3 were formed by exon-skipping in the 3'-untranslated region (UTR), while V4 was formed by alternative 3' splice site of exon 3, resulting in a premature termination codon, combined with exon skipping in the 3'-UTR. Expression profile analysis showed that all the isoforms were most abundant in the spleen, and V1 was strongly induced by poly(I:C). Furthermore, the transcription of V1 altered with the increasing age and differed between species. Exon skipping in the 3'-UTR of V2 and V3 down-regulated expression of the luciferase reporter gene, and hence presumably of the PILRB gene, while V4 was subjected to nonsense-mediated mRNA decay. Additionally, five novel splicing patterns were detected using the minigene approach, indicating complex AS of porcine PILRB. These results will help to reveal the role of PILRß in the host immune response using pig models, and will facilitate the breeding of pigs resistant to viral diseases through molecular breeding methods.

Electrochemical formation of Mg-Li-Ca alloys by codeposition of Mg, Li and Ca from LiCl-KCl-MgCl2-CaCl2 melts.

This work presents electrochemical formation of Mg-Li-Ca alloys via codeposition of Mg, Li and Ca on a molybdenum electrode in KCl-LiCl-MgCl(2)-CaCl(2) melts at 943 K. Cyclic voltammograms (CVs) showed that the underpotential deposition (UPD) of calcium on pre-deposited magnesium leads to the formation of a liquid Mg-Ca alloy, and the succeeding underpotential deposition of lithium on pre-deposited Mg-Ca alloy leads to the formation of a liquid Mg-Li-Ca solution. Chronopotentiometric measurements indicated that the codepositon of Mg, Li and Ca occurs at current densities more negative than -0.31 A cm(-2) in LiCl-KCl-MgCl(2) (5 wt%) melts containing 1 wt% CaCl(2). Chronoamperograms demonstrated that the onset potential for the codeposition of Mg, Li and Ca is -2.200 V, and the codeposition of Mg, Li and Ca is formed when the applied potentials are more negative than -2.200 V. X-Ray diffraction (XRD) indicated that Mg-Li-Ca alloys with different phases were formed via galvanostatic electrolysis. The microstructures of typical alpha and beta phases of Mg-Li-Ca alloys were characterized by optical microscope (OM) and scanning electron microscopy (SEM). The analysis of energy dispersive spectrometry (EDS) showed that the element Ca mainly distributes along grain boundary in Mg-Li-Ca alloys. The results of inductively coupled plasma analysis determined that the chemical compositions of Mg-Li-Ca alloys correspond with the phase structures of XRD patterns, and the lithium and calcium contents of Mg-Li-Ca alloys depend on the concentrations of MgCl(2) and CaCl(2).

Establishment of a purificatory method for alpha-fetoprotein variant by affinity adsorption / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To establish a purificatory method of alpha-fetoprotein variant (AFP-L3) based on microspincolumn with lens culinaris agglutinin (LCA).</p><p><b>METHODS</b>LCA was isolated by ammonium sulfate precipitation method from lens culinaris. AFP-L3 affinity adsorption microspincolumns which were made from LCA coupled with activated Sepharose 4B were prepared. By adding into the centrifuge column, serum was absorbed and eluted to purify AFP-L3. The results of purified AFP-L3 detection of 10 cases AFP positive sera by electro-chemiluminescence immunoassay were compared with traditional crossed affinity immunoelectrophoresis.</p><p><b>RESULTS</b>8 of 10 cases AFP-L3 concentration were greater than 5 ng/ml in purified sera. Six cases show positive reaction in affinity immune cross electrophoresis experiment.</p><p><b>CONCLUSION</b>Successfully established purification method of AFP-L3 by affinity absorption based on microspincolumn. The method was more conducive to clinical laboratory applications due to its high sensitive and easy operation.</p>