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Objectives The effect of marital status on the prognosis of intrahepatic cholangiocarcinoma (ICC) has not been strictly studied. Therefore, the present study focused on exploring whether marital status was related to ICC prognosis. Methods This study chose cases from Surveillance, Epidemiology, and End Results (SEER) database during the 2004–2015 period. Cancer-specific survival (CSS) and overall survival (OS) were compared between married and unmarried groups. Results We enrolled 4550 eligible cases, among them, 2760 were married (60.66%). Married subjects had more younger age (P P P P = 0.003] as well as OS (HR: 0.903; 95%CI: 0.845–0.965; P = 0.003). Upon subgroup analysis, married cases had prolonged CSS as well as OS to unmarried cases of almost every subgroup, especially for patients who have not received surgery, chemotherapy, or radiotherapy. Conclusion This study identified marital status as the factor independently predicted ICC prognosis. The dismal prognostic outcome of unmarried cases may attribute to inadequate treatments, socioeconomic and psychosocial factors.

The myocyte enhancer factor 2C (MEF2C) is a member of the MEF2 family of transcription factors, involved in skeletal muscle development. In this study we report the cDNA sequence and isolate the 5' upstream region of the mef2c gene from porcine genomic DNA using PCR-based GenomeWalker. The open reading frame of porcine mef2c cDNA covers 1,392 bases, encoding 464 amino acids, which show 94% identity with human MEF2C at the level of the primary protein structure. Annear the C terminus of mef2c, a 96-nt sequence appear to represent alternatively spliced transcripts was present in some cDNAs and absent in the other. No typical TATA, GC box or CAAT box binding site was found in porcine mef2c 5' upstream region, whereas some potential binding sites for MyoD (E-box), MEF2 and MBF1 were present in the proximal upstream region. Transfection of the mef2c 5' upstream region with EGFP into cos7 cells demonstrated that the region from -162 to +115 bp immediately 5' of the exon 1 was sufficient to direct strong EGFP protein expression. Co-transfection assays demonstrated that MBF1 bound the mef2c promoter and inhibited mef2c expression. These results may be useful for elucidating the regulation mechanisms of mef2c, which interacts with other factors to regulate target genes.

The protein encoded by SLC27A2 gene is an isozyme of long-chain fatty-acid-coenzyme A ligase family, and it converts free long-chain fatty acids into fatty acyl-CoA esters, and thereby plays a key role in lipid biosynthesis and fatty acid degradation. In the present study, SLC27A2 located on human chromosome 15 was selected as candidate gene and we isolated and cloned partial fragments of mRNA sequence and genomic fragments of porcine SLC27A2 gene. The coding region of the gene as determined by alignments shared 90% and 82% identity with human and mouse cDNAs, respectively. Detection in LargeWhite and Meishan breeds showed that a single nucleotide polymorphism (SNP) (A→G) existed in exon 7, which caused corresponding amino acid changed for encoding. In LargeWhite pigs it encoded for Val while in Meishan pigs it encoded for Ile, so we developed the PCR-RFLP genotype method for detection of this polymorphism. Association study in 135 F 2 reference family indicated that significant correlation existed between the polymorphism and growth and carcass traits.

To investigate the molecular basis of porcine heterosis, suppression subtractive hybridization (SSH) was performed to detect the differences in gene expression between porcine longissirnus dorsi of Meishan × Large White (MS × LW) F, hybrids and their parents Meishan pigs. An expression sequence tag (EST) differentially expressed was found, designated as ML556, which was homologous to a hypothetical protein HSPC117, from human hematopoietic stedprogenitor cells (HSPCs), and the full-length cDNA of porcine HSPC117 was cloned using rapid amplification of cDNA ends (RACE) method. Translation of the mRNA transcript revealed an open reading frame (ORF) of 505 amino acid residues encoding a peroxisomal targeting signal (PTS) with theoretical molecular weight of 55 kDa. Alignment analysis revealed that the deduced protein sequence exhibit 98, 98, 98, 97, and 97% identity with that of cattle, human, dog, rat, and mouse, respectively. The tissue expression analysis indicated that the porcine HSPC117 gene is highly expressed in muscle, spleen, lung, kidney, uterus, ovary and testis, moderately expressed in fat, heart, and liver, and not expressed in stomach and small intestine. The possible role of porcine HSPC117 and its relationship with porcine heterosis were discussed.

mRNA differential display technique was performed to discuss the differential expression of genes in fat tissue between introduced European and Chinese indigenous pigs. Four anchor primers in combination with five arbitrary primers (20 sets in total) were used and nearly 300 bands were observed in polyacrylamide gel, among which 29 differential display bands were obtained. Twelve of 29 cDNA fragments were identified using reverse Northern dot blot, and subsequently cloned and sequenced. Eight of 12 cDNAs had no matches in GenBank and were submitted to GenBank, and the other 4 showed similarity to identified genes from GenBank. Three among 8 novel ESTs were selected to be further identified by semi-quantitative RT-PCR. In our experiment, silver staining DDRT-PCR and DIG primer DNA labeling reverse Northern dot blot were used to avoid radioactive pollution. The result showed that the expressions of 5 among 8 novel ESTs were stronger in the backfat of Tongcheng pigs and the others were weaker than that in Duroc pigs. These novel ESTs were prepared for selecting genes related to adipose cells.

In pig industry, fat deposition related traits such as back fat thickness and fat rate are of great economic importance. Thus, research on genes related with fat deposition can offer many useful values theoretically and practically. Gene FIT1 (Fat-inducing transcript 1) plays an important role in packaging lipid droplets. Here, we used FIT1 gene as the candidate gene for fat deposition. Sequence comparison revealed that an insertion/deletion mutation occurred at 590~595 bp of the second exon. We then carried out PCR-SSCP analysis followed by association analysis in F2 "Large white xMeishan" resource family. In all the individuals tested, all Meishan pigs possessed the insertion, which was designated allele A, while most Large white pigs possessed the deletion and was named as allele B. Association analysis in F2 resource family showed that this site was highly associated with fat percentage (FP), 6-7 rib fat thickness (RFT), buttock fat thickness (BFT), leaf fat weigh (LFW), total internal fat weigh (TFW), and internal fat rate (IFR) (Plt;0.01). These results indicated that FIT1 gene may have some important values for application. Further and deep research is necessary for revealing more information on this gene in order to provide a new marker for molecular marker-assisted selection breeding.

In order to detect the molecular basis of heterosis in pigs, suppression subtractive hybridization was carried out to investigate the difference in gene expression in the Longissimus dorsi muscle tissues between MeishanxYorkshire F1 crossbreeds and their parents, Meishan pigs. The swine myosin regulatory light chain 2 (MRLC2) gene differentially expressed between the crossbreeds and the purebreds was isolated and identified using semi-quantitative reverse transcriptase polymerase chain reaction and its complete cDNA sequence was obtained using the rapid amplification of cDNA ends method. The nucleotide sequence of the gene is not homologous to any of the known porcine genes. The sequence prediction analysis reveals that the open reading frame of this gene encodes a protein of 172 amino acids containing the putative conserved domain of the EF-hand superfamily. This predicted amino acid sequence of porcine MRLC2 protein exhibits 99%, 98%, 98%, 98% and 97% identity with that of cattle, human, dog, rat and mouse, respectively. The homology analysis revealed that the MRLC2 protein was very much conserved in evolution. The tissue expression analysis indicated that the swine MRLC2 gene is highly expressed in muscle, fat, heart, liver, spleen, lung, kidney, stomach, small intestine, ovary and testis, but not expressed in pancreas.

To study the molecular basis of heterosis, suppression subtractive hybridization was used to investigate the differences in gene expression between porcine Longissimus dorsi of F1 hybrids Large WhitexMeishan and their female parents Meishan. From two specific subtractive cDNA libraries, the clones selected by reverse Northern high-density blot screening were chosen to clone full-length cDNA by rapid amplification of cDNA ends. An expression-upregulated gene for Meishan skeletal muscle, designated protein phosphatase 1, catalytic subunit, beta isoform (PPP1CB), was identified. Porcine PPP1CB contains an open reading frame encoding 327 amino acid residues with 13 and 1763 nucleotides in the 5' and 3' untranslated regions, respectively. A DNA fragment of 721 nucleotides was amplified and a mutation that creates/disrupts a restriction site for endonuclease RsaI was found. The derived amino acid sequence of PPP1CB has high homology with the PPP1CB of three species, Mus musculus (99%), human (99%) and mouse (100%). The tissue expression analysis indicated that the swine PPP1CB gene is generally expressed in most tissues. The possible role of PPP1CB and its relation to porcine heterosis are discussed.

Using suppression subtractive hybridization (SSH) technique, forward and reverse subtracted cDNA libraries were constructed between Longissimus muscles from Meishan and Landrace pigs. A housekeeping gene, G3PDH, was used to estimate the efficiency of subtractive cDNA. In two cDNA libraries, G3PDH was subtracted very efficiently at appropriate 2(10) and 2(5) folds, respectively, indicating that some differentially expressed genes were also enriched at the same folds and the two subtractive cDNA libraries were very successful. A total of 709 and 673 positive clones were isolated from forward and reverse subtracted cDNA libraries, respectively. Analysis of PCR showed that most of all plasmids in the clones contained 150-750 bp inserts. The construction of subtractive cDNA libraries between muscle tissue from different pig breeds laid solid foundations for isolating and identifying the genes determining muscle growth and meat quality, which will be important to understand the mechanism of muscle growth, determination of meat quality and practice of molecular breeding.