Development and characterization of microsatellite markers for Solen grandis using Illumina sequencing approach.

Solen grandis is an important economic and overexploited bivalve species. In order to perform its fine-scale genetic analyses, 105 pairs of microsatellites with polymorphism were identified through Illumina Hiseq platform and bioinformatic assembly technology in this study. The estimated fragment size ranged from 100 to 268 bp and the number of alleles per locus varied between 2 and 23. Observed and expected heterozygosities varied from 0.0667 to 1.0000 and 0.0966 to 0.9492, respectively. Fourteen loci deviated significantly from Hardy-Weinberg equilibrium after Bonferroni correction. These microsatellite markers developed in this study would be helpful for future genetic studies on S. grandis and closely related species.

Invasive pulmonary fungal infections in patients with connective tissue disease: a retrospective study from northern China.

Invasive pulmonary fungal infection (IPFI) is a potentially fatal complication in patients with connective tissue disease (CTD). The current study aimed to uncover the clinical characteristics and risk factors of patients with IPFI-CTD. The files of 2186 CTD patients admitted to a single center in northern China between January 2011 and December 2013 were retrospectively reviewed. A total of 47 CTD patients with IPFI were enrolled into this study and assigned to the CTD-IPFI group, while 47 uninfected CTD patients were assigned to the control group. Clinical manifestations were recorded, and risk factors of IPFI were calculated by stepwise logistical regression analysis. Forty-seven (2.15%) CTD patients developed IPFI. Systemic lupus erythematosus patients were responsible for the highest proportion (36.17%) of cases with IPFI. Candida albicans (72.3%) accounted for the most common fungal species. CTD-IPFI patients had significantly elevated white blood cell count, erythrocyte sedimentation rate, C-reactive protein and fasting glucose values compared to controls (P<0.05). Cough, sputum and blood in phlegm were the most common symptoms. Risk factors of IPFI in CTD included maximum prednisone dose ≥30 mg/day within 3 months prior to infection, anti-microbial drug therapy, and interstitial pneumonia. CTD patients who have underlying interstitial pneumonia, prior prednisone or multiple antibiotics, were more likely to develop IPFI.

Comparative RNA profile analysis of idiopathic dilated cardiomyopathy and ischemic cardiomyopathy.

Previous research has focused on revealing the functions of each individual gene and/or pathway in idiopathic dilated cardiomyopathy (DCM) or ischemic cardiomyopathy (IC). However, the common or specific pathways of the initiation and processes of DCM and IC are still unclear. Here, we attempted to uncover the critical genes and potential molecular networks that play important roles in DCM and IC progression commonly or specifically. The transcriptional profiles from normal and DCM or IC patient samples were analyzed and compared using bioinformatic methods. Initially, the normal and DCM or IC sample data were processed and the most notable differentially expressed genes (DEGs) from DCM or IC were identified. By comparing the DEGs from DCM with those from IC, the DCM- and IC-specific DEGs were identified. The gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses indicated the significance of multiple biological processes as well as signaling pathways that affect heart function and DCM or IC progression. Protein-protein interaction network analysis identified the relationships between different genes, and some important genes such as MYC and FN1 were found to be hubs, which master each individual module of DCM-specific and IC-specific DEGs, respectively. We discovered commonalities and differences of gene expression profiles and molecular pathways between different cardiomyopathies. The gene discovery and molecular signature analysis in this study could offer insights into disease mechanisms and also identify markers useful for diagnostic, prognostic, and therapeutic purposes.

Invasive pulmonary fungal infections in patients with connective tissue disease: a retrospective study from northern China

Invasive pulmonary fungal infection (IPFI) is a potentially fatal complication in patients with connective tissue disease (CTD). The current study aimed to uncover the clinical characteristics and risk factors of patients with IPFI-CTD. The files of 2186 CTD patients admitted to a single center in northern China between January 2011 and December 2013 were retrospectively reviewed. A total of 47 CTD patients with IPFI were enrolled into this study and assigned to the CTD-IPFI group, while 47 uninfected CTD patients were assigned to the control group. Clinical manifestations were recorded, and risk factors of IPFI were calculated by stepwise logistical regression analysis. Forty-seven (2.15%) CTD patients developed IPFI. Systemic lupus erythematosus patients were responsible for the highest proportion (36.17%) of cases with IPFI. Candida albicans (72.3%) accounted for the most common fungal species. CTD-IPFI patients had significantly elevated white blood cell count, erythrocyte sedimentation rate, C-reactive protein and fasting glucose values compared to controls (P<0.05). Cough, sputum and blood in phlegm were the most common symptoms. Risk factors of IPFI in CTD included maximum prednisone dose ≥30 mg/day within 3 months prior to infection, anti-microbial drug therapy, and interstitial pneumonia. CTD patients who have underlying interstitial pneumonia, prior prednisone or multiple antibiotics, were more likely to develop IPFI.

Genomic survey and gene expression analysis of the VDAC gene family in rice.

The voltage-dependent anion channel (VDAC), also known as a mitochondrial porin, plays an important role in the regulation of metabolic and energetic functions of mitochondria, as well as in mitochondria-mediated apoptosis. Cytoplasmic male sterility (CMS) is of major economic importance for commercial hybrid production and a research model for the interaction be-tween nuclear and cytoplasmic genomes. Recent research has revealed that CMS is associated with programmed cell death. Here, we used the Honglian (HL)-CMS line of rice (Oryza sativa) as material to investigate the association of O. sativa VDAC (OsVDAC) expression to CMS. Eight VDACs were extracted from rice in this study. Bioinformatic analysis of the rice VDACs was conducted at the DNA, cDNA, and protein level. Expression patterns of OsVDACs were analyzed in different organs and during different stages of pollen development using sterile line YuetaiA (YTA), and its maintainer line YuetaiB (YTB). Differential expression of OsVDACs between YTA and YTB was observed, suggesting that VDACs may be involved in the formation of HL-CMS.

Analysis of the drug-resistant characteristics of Klebsiella pneumoniae isolated from the respiratory tract and CTX-M ESBL genes.

The main aim of this study was to understand the relationship between the drug-resistant characteristics of Klebsiella pneumoniae and CTX-M-type extended spectrum ß-lactamases (ESBLs), and to detect the distributions of CTX-M-type ESBLs in clinically isolated strains. CTX-M ESBL genes isolated from the clinical samples were amplified by polymerase chain reaction and identified by sequence analysis; the antibiotic susceptibility of the samples was determined using the Kirby-Bauer disc-diffusion method. One hundred and five strains among the 246 isolated strains of K. pneumoniae tested positive for ESBL production (42.68%); 92 of these produced CTX-M ESBLs. Of the 92 CTX-M ESBL strains, 81 produced CTX-M-1 ESBLs and 11 produced CTX-M-25 ESBLs. Fifty-seven of the CTX-M-1 ESBL- and six of the CTX-M-25 ESBL-producing bacteria had CTX-M ESBL genes that coexisted in the plasmid and chromosome. The Kirby-Bauer antibiotic susceptibility method revealed that CTX-M ESBL-positive strains showed a higher rate of resistance to cefazolin, cefoxitin, cefuroxime, ceftazidime, cefotaxime, aztreonam, levofloxacin, and cotrimoxazole, compared to the CTX-M ESBL-negative strains (P < 0.05). The CTX-M ESBL genes were commonly observed in the K. pneumoniae isolated from respiratory tract samples; these were significantly associated with the drug-resistant characteristics of K. pneumoniae to ß-lactam antibiotics.

rbcS SRS4 promoter from Glycine max and its expression activity in transgenic tobacco.

The regulatory region of the ribulose-1,5-bisphosphate carboxylase small subunit gene SRS4 from soybean (Glycine max) was cloned using TAIL-PCR and general PCR, and named the rbcS promoter. The promoter was fused with the GUS gene and introduced into Nicotiana tabacum via Agrobacterium-mediated leaf disk transformation. In 4-week-old transgenic tobacco plants, the highest GUS expression levels were observed in the leaves, GUS activity was 7.13- and 7.40-fold higher in leaves than in stems and roots, respectively. Moreover, GUS activity was stimulated by light. In conclusion, spatial and light regulation of the soybean rbcS promoter was observed in N. tabacum, thus illustrating a leaf-specific and light-induced promoter.

Long-term, regular remote ischemic preconditioning improves endothelial function in patients with coronary heart disease

Remote ischemic preconditioning (RIPre) can prevent myocardial injury. The purpose of this study was to assess the beneficial effects of long-term regular RIPre on human arteries. Forty patients scheduled for coronary artery bypass graft (CABG) surgery were assigned randomly to a RIPre group (n=20) or coronary heart disease (CHD) group (n=20). Twenty patients scheduled for mastectomy were enrolled as a control group. RIPre was achieved by occluding arterial blood flow 5 min with a mercury sphygmomanometer followed by a 5-min reperfusion period, and this was repeated 4 times. The RIPre procedure was repeated 3 times a day for 20 days. In all patients, arterial fragments discarded during surgery were collected to evaluate endothelial function by flow-mediated dilation (FMD), CD34+ monocyte count, and endothelial nitric oxide synthase (eNOS expression). Phosphorylation levels of STAT-3 and Akt were also assayed to explore the underlying mechanisms. Compared with the CHD group, long-term regular RIPre significantly improved FMD after 20 days (8.5±2.4 vs 4.9±4.2%, P<0.05) and significantly reduced troponin after CABG surgery (0.72±0.31 and 1.64±0.19, P<0.05). RIPre activated STAT-3 and increased CD34+ endothelial progenitor cell counts found in arteries. Long-term, regular RIPre improved endothelial function in patients with CHD, possibly due to STAT-3 activation, and this may have led to an increase in endothelial progenitor cells.

Expression of ß-defensins in intestines of chickens injected with vitamin D3 and lipopolysaccharide.

The objective of this study was to evaluate the effect of vitamin D3 (VD3) on the regulation of chicken intestinal ß-defensin genes under normal and lipopolysaccharides (LPS) conditions. Four treatment groups were used, including a negative control group, VD3-injection group, LPS-injection group, and both VD3-injection and LPS-injection group. At 4, 24, and 48 h post-injection, intestines were collected and RNA was isolated to measure the chicken ß-defensin genes with putative vitamin D responsive elements using quantitative polymerase chain reaction. Expressions of all 7 chicken ß-defensin genes was detectable in the intestines. Significant increases in GAL-6, -7 and -9 were found following LPS injection treatment at 4, 24, and 48 h post-injection, respectively, whereas VD3 injection did not affect the expression of any investigated genes under normal conditions. However, the expression of GAL-4, -5, -6, and -10 were synergistically upregulated by VD3 in combination with LPS. These results suggest that VD3 enhances the immune immunity during LPS challenge by inducing the expression of chicken ß-defensin genes when birds are exposed to immune stressors.

Long-term, regular remote ischemic preconditioning improves endothelial function in patients with coronary heart disease.

Remote ischemic preconditioning (RIPre) can prevent myocardial injury. The purpose of this study was to assess the beneficial effects of long-term regular RIPre on human arteries. Forty patients scheduled for coronary artery bypass graft (CABG) surgery were assigned randomly to a RIPre group (n=20) or coronary heart disease (CHD) group (n=20). Twenty patients scheduled for mastectomy were enrolled as a control group. RIPre was achieved by occluding arterial blood flow 5 min with a mercury sphygmomanometer followed by a 5-min reperfusion period, and this was repeated 4 times. The RIPre procedure was repeated 3 times a day for 20 days. In all patients, arterial fragments discarded during surgery were collected to evaluate endothelial function by flow-mediated dilation (FMD), CD34(+) monocyte count, and endothelial nitric oxide synthase (eNOS expression). Phosphorylation levels of STAT-3 and Akt were also assayed to explore the underlying mechanisms. Compared with the CHD group, long-term regular RIPre significantly improved FMD after 20 days (8.5±2.4 vs 4.9±4.2%, P<0.05) and significantly reduced troponin after CABG surgery (0.72±0.31 and 1.64±0.19, P<0.05). RIPre activated STAT-3 and increased CD34(+) endothelial progenitor cell counts found in arteries. Long-term, regular RIPre improved endothelial function in patients with CHD, possibly due to STAT-3 activation, and this may have led to an increase in endothelial progenitor cells.

Polymorphic microsatellite loci for the razor clam, Sinonovacula constricta.

The razor clam, Sinonovacula constricta, is an important commercial bivalve and a popular mollusca food in China. Twelve polymorphic microsatellite markers were isolated from the razor clam using a partial genomic library enriched for tandem repeat sequences of (CA)16, (GA)16. Polymorphisms of these loci were evaluated in a wild population of 30 individuals. The allele number of these polymorphic markers ranged from 5-15 per locus with an average of 9.333. Observed and expected heterozygosity values ranged from 0.192-1.000 and 0.219-0.906. Polymorphism information content ranged from 0.209-0.892 with an average of 0.704. Three loci significantly deviated from Hardy-Weinberg equilibrium after Bonferroni correction. No significant linkage disequilibrium was detected between these loci. This set of microsatellite loci are useful for genetic studies in S. constricta.

Improvement in the carcass traits and meat quality of growing-finishing Rongchang pigs by conjugated linoleic acid through altered gene expression of muscle fiber types.

A total of 160 Rongchang pigs (26.76±1.78 kg) were randomly assigned to 5 dietary treatment groups until their body weight (BW) reached 90 kg. The diets were supplemented with 0, 0.5, 1.0, 1.5, and 2.0% conjugated linoleic acid (CLA). Our results showed that the 1.0 to 2.0% CLA-fed pigs had less back fat deposition when their BW reached 90 kg than the pigs that received less than 1% CLA. During the 30 to 60 kg growing period, 1.0, 1.5, and 2.0% CLA treatments improved pork quality by significantly reducing the pork pH (P<0.01) and color value (P<0.05), but they increased marble scaling (P<0.01). Similarly, the 1.5 and 2.0% CLA-fed pigs had more marble than other pigs when their BW reached 90 kg. Furthermore, CLA significantly affected the expression of muscle fiber-type genes. The 1.5% CLA-fed pigs exhibited the highest mRNA expression of MyHC1 and MyHC2a (P<0.05) at 60 kg BW. At 90 kg BW, the highest expression of MyHC1 and MyHC2a (P<0.05) was found in the 2.0% CLA group. However, MyHC2x was downregulated in the CLA-fed pigs at this time. In addition, CLA supplements did not evidently alter mRNA expression of MyHC2b at all times. These results demonstrate that CLA could affect carcass traits and improve the meat quality of growing-finishing pigs by altering the expression of genes related to muscle growth and development; 1-1.5% CLA was the most appropriate CLA dose.

Lack of association of IL-2RA and IL-2RB polymorphisms with rheumatoid arthritis in a Han Chinese population.

Polymorphisms in IL-2RA and IL-2RB genes have been reported to confer susceptibility to rheumatoid arthritis (RA) in European populations. We investigated a possilbe association between SNPs in IL-2RA and IL-2RB genes and RA in a Han Chinese population. rs2104286 in IL-2RA and rs743777 in IL-2RB genes were genotyped in a Han Chinese cohort composed of 500 patients with RA and 600 controls. The levels of anti-cyclic citrullinated peptide antibodies (CCP) and rheumatoid factor were determined in all patients and controls. The genotype and allele frequencies of the two SNPs were compared in patients and controls. Additionally, serum concentrations of anti-CCP and rheumatoid factor were analyzed in the three genotype groups of IL-2RA and IL-2RB genes. There was no overall difference in the genotype and allele frequencies of the two SNPs, rs2104286 in IL-2RA and rs743777 in IL-2RB, between the patients with RA and controls. In addition, none of the subgroups showed any significant association with RA risk after stratification by CCP and rheumatoid factor levels. We conclude that the two genetic variants within IL-2RA and IL-2RB are not associated with genetic susceptibility to RA in Han Chinese. Also, the rs2104286 and rs743777 genotypes were not significantly associated with the concentrations of anti-CCP antibodies or rheumatoid factor.

Transcriptional activity of an ovarian-specific promoter from rat in dairy goat granulosa cells.

Ovarian-specific promoter 1 (OSP-1) is a retrovirus-like element isolated from the complementary DNA library of rat that has been thought to be specifically expressed in ovary. To exploit this promoter in dairy goat ovary granulosa cells (GCs), OSP-1 from rat was used to construct the reporter vector pOSP-1-EGFP, in which egfp coding for enhanced green fluorescent protein (EGFP) was used as a reporter to examine the activity of OSP-1 in GCs. EGFP was successfully expressed in dairy goat GCs transfected with pOSP-1-EGFP. Reverse transcriptase-polymerase chain reaction analysis confirmed the tissue-specific transcription of EGFP messenger RNA in dairy goat GCs transfected with pOSP-1-EGFP. We concluded that OSP-1 promoter from rat can specifically drive foreign gene expression in dairy goat GCs. Thus, we obtained a tissue-specific regulation element and provided a potential tool for the research of regulation and development of the ovary in dairy goats.