Identification and Expression Profiling of Odorant Receptor Protein Genes in Sitophilus zeamais (Coleoptera: Curculionoidea) Using RT-qPCR.

This study aimed to identify ORs (odorant receptors) and Orco (odorant receptor coreceptor) genes in Sitophilus zeamais Motschulsky (Coleoptera: Curculionoidea), to explore the relative expression levels of these genes in different adult tissues and obtain information on highly expressed receptor proteins. Putative OR and Orco genes were identified from transcriptomic data previously obtained for S. zeamais using bioinformatics methods. Quantitative real-time PCR was used to compare the differences in expression in seven adult tissues (male antennae, female antennae, heads, thoraxes, abdomens, wings, and legs). The candidate OR and Orco gene sequences were analyzed, and the protein physicochemical properties were predicted. We identified 64 OR genes including the Orco gene. Forty-seven OR genes, including Orco, were over expressed in male or female antennae. Seventeen OR genes appeared to be expressed at elevated levels in male antennae. Twenty-nine genes were expressed at significantly elevated levels in female antennae. In total, 11 OR genes were selected for further sequence analysis. The selected proteins were structurally characterized, and bioinformatics analysis was performed. Overall, in this study, candidate ORs of S. zeamais have been identified for the first time, and these ORs could be molecular targets for interference in the insect olfactory system.

Characterization of novel polymorphic genomic microsatellite markers of Boehmeria tricuspis (Hance) Makino.

In the present study, 59 polymorphic microsatellite loci of Boehmeria tricuspis (Hance) Makino were developed from the specific length amplified fragment sequencing data library of genome. The number of alleles per locus ranged from two to five, and the observed and expected heterozygosities ranged from 0.0000 to 1.0000 and from 0.0769 to 0.6751, respectively. Among the 59 loci, 25 displayed significant deviations from Hardy-Weinberg expectations (P < 0.05). The developed simple sequence repeat markers should be useful for studying population genetics in B. tricuspis (Hance) Makino, for providing further knowledge on its population differentiation, breeding system, and dispersal ability, as well as quantitative trait locus mapping. These markers could also be valuable genetic resources for closely related species.

Protein-protein interaction network and mechanism analysis of hepatitis C.

We predicted potential genes and identified pathways associated with hepatitis C. The gene expression profiles of GSE40184 from blood samples and GSE38597 from liver biopsy samples were downloaded from the GEO database. Differentially expressed genes (DEGs) were recognized using the Limma Package. The Pearson correlation test was used to construct the co-expression network of DEGs. Gene set enrichment analysis was used to define significant functions and pathways for DEGs. A total of 165 DEGs in blood samples and 523 DEGs in liver biopsy samples were identified. Eight DEGs were common between these samples. Gene Ontology enrichment analysis showed that 165 DEGs in blood samples were significantly enriched regarding the response to protein binding, receptor binding, G-protein coupled receptor binding, cytokine receptor binding, and cytokine activity. The most significant term of the Kyoto Encyclopedia of Genes and Genomes pathway was the cytokine-cytokine receptor interaction. Protein-protein interaction network analysis indicated that three subnetworks with more nodes and edges were involved in these interactions. We used robust biomarkers that were differentially expressed in hepatitis C and determined their relevance in the biological function, signal pathways, protein-protein interaction network, and co-expression network of hepatitis C.

Effectiveness of microsatellite and single nucleotide polymorphism markers for parentage analysis in European domestic pigs.

Parentage analysis and individual identification are recent, promising methods that have been applied to evolutionary and ecological studies, as well as conservation management. Parental exclusion relying on polymorphic microsatellites has been used worldwide in parentage determination, while the low mutation rate and genotyping error rate of single nucleotide polymorphisms (SNPs) make them another important marker for pedigree tracing. Here, we compared the effectiveness of microsatellites and SNP markers in European pigs. We also measured and presented the minimum and optimal criteria for SNP markers to be used in paternity and identity analysis. Our findings may contribute to the development of techniques for future molecular evolution and conservation studies, as well as breeding programs.

miR-96 inhibits cardiac hypertrophy by targeting growth factor receptor-bound 2.

Increasing evidence has indicated that microRNAs are involved in the pathogenesis of cardiac hypertrophy. However, whether miR-96 is involved in heart diseases, particularly cardiac hypertrophy, remains unclear. In this study, we found that miR-96 is a negative regulator of cardiac hypertrophy. In primary cardiomyocytes, overexpression of miR-96 inhibited phenylephrine-induced cardiomyocyte hypertrophy and decreased the mRNA expression of cardiac hypertrophy markers such as atrial natriuretic factor and ß-myosin heavy chain. Interestingly, we found that growth factor receptor-bound 2 is a direct target of miR-96, which is a negative regulator of cardiac hypertrophy. Overexpression of miR-96 in cardiomyocytes led to reduced growth factor receptor-bound 2 expression. More importantly, miR-96 repressed the extracellular-regulated protein kinase signaling pathway by targeting growth factor receptor-bound 2 in cardiomyocytes. Our data demonstrate that miR-96 is a negative regulator of cardiac hypertrophy and extracellular-regulated protein kinase signaling, thus offering a new therapeutic strategy for cardiac hypertrophy.

Analysis of age of onset, pre-existing infections, and features of magnetic resonance imaging results in patients with acute myelitis.

The clinical features and potential risk factors of acute myelitis (AM) were investigated. The medical records of patients with AM admitted to our department between January 2004 and December 2011 were collected and retrospectively analyzed. The diagnosis of AM was in line with the diagnostic criteria of the Transverse Myelitus Consortium Working Group. The age of onset, clinical, and imaging features of these patients were analyzed. A total of 64 patients satisfying the inclusion criteria were enrolled in the study, including 39 males and 25 females. The patients ranged in age from 1 to 80 years, with a mean age of 34 years. Twenty-three patients had symptoms of pre-existing infections. The correlation between spinal cord lesions and spinal lesions was statistically significant (P<0.05). Cervical spinal cord inflammation was the most common. Prodromal infections were more commonly observed in thoracic spinal cord myelitis than in cervical spinal cord myelitis (P<0.05). AM appears to be more likely to occur in male minors. Lesions of the spinal column were partially implicated with the occurrence of myelitis, which suggests that such lesions might be a predisposing factor. Compared to AM of the cervical cord, pre-existing infections appear to be of greater significance for the occurrence of myelitis of the thoracic cord.

Effect of the IkBα mutant gene delivery to mesenchymal stem cells on rat chronic pancreatitis.

This study aimed to investigate the effect of inhibitors of the NF-kΒ alpha mutant gene (IkBaM) delivery to mensenchymal stem cells (MSCs) on rat chronic pancreatitis (CP). A total of 120 Sprague-Dawley rats were randomly divided into 6 groups of 20: Group A was injected with sterile saline solution, Group B was injected with allogenic MSCs, Group C1 was injected with allogenic IkBαM-MSCs cultured in vitro 4 h before CP modeling, Group C2 was injected with allogenic IkBαM-MSCs cultured in vitro during CP modeling, Group C3 was cultured with allogenic IkBαM-MSCs cultured in vitro 4 h after CP modeling, and Group D was injected with rAAV2-MSCs. Cytokine levels of ICAM-1, CTGF, IL-1, IL-6, IL-8, TNF-α, TIMP-1, TIMP-2, IL-10, FN, MMP-1, MMP-2, MMP-3, and MMP-9 were examined. The results indicated that allogenic IκBαM-MSCs could reduce pro-inflammatory cytokine levels and increase anti-inflammatory cytokine levels in CP. The allogenic IkBαM-MSCs reduced the activation and promoted the apoptosis of pancreatic stellate cells in the rat model of CP. IkBαM-MSCs influenced the proliferation and apoptosis of pancreatic stellate cells by regulating the activation of the PPAR, MAPK, mTOR, TGF-ß, NOD-like receptor, Notch, WNT, TGF-ß1-SMAD-2/3, and P53 signal transduction pathways.

Effect of atorvastatin on left atrial function of patients with paroxysmal atrial fibrillation.

The aim of this study was to evaluate the effects of atorvastatin on left atrial (LA) function in paroxysmal atrial fibrillation patients. Fifty-eight paroxysmal atrial fibrillation patients were divided into two groups (treatment and control groups). The echocardiography parameters, including LA active emptying volume (LAAEV), LA active emptying fraction, LA maximum volume, LA total emptying volume, LA total emptying fraction, and LA ejection force (LAEF), were measured before treatment, and then 12 and 18 months after treatment. Compared to pre-treatment levels, the parameters reflecting LA pump function, such as LAAEV and LAEF, decreased significantly in treatment groups 12 months after treatment (P < 0.05). LAAEV and LAEF significantly increased 18 months after treatment (P < 0.05), and the indicators reflecting LA reservoir function, such as maximum volume, total emptying volume, and total emptying fraction increased significantly 18 months after treatment (P < 0.05). Compared with pre-treatment levels, LAAEV and LAEF decreased significantly 18 months after treatment in the control group (P < 0.05). These results demonstrated that long-term atorvastatin treatment could ameliorate the function of the atrium sinistrum.

Polymorphic microsatellite loci for Japanese Spanish mackerel (Scomberomorus niphonius).

We isolated and characterized 21 polymorphic microsatellite loci in Japanese Spanish mackerel (Scomberomorus niphonius) using a (GT)(13)-enriched genomic library. Forty individuals were collected from Qingdao, China. We found 3 to 24 alleles per locus, with a mean of 8.8. The observed and expected heterozygosities ranged from 0.263 to 0.975 and from 0.385 to 0.946, with means of 0.655 and 0.685, respectively. Deviation from Hardy-Weinberg proportions was detected at three loci. Two loci showed evidence for null alleles. These microsatellite markers will be useful for population genetic analysis of Japanese Spanish mackerel.

Genomic structure, polymorphism and expression analysis of growth hormone-releasing hormone and pituitary adenylate cyclase activating polypeptide genes in the half-smooth tongue sole (Cynoglossus semilaevis).

Growth hormone-releasing hormone (GHRH) and pituitary adenylate cyclase activating polypeptide (PACAP) regulate development and somatic growth in teleosts; they may be associated with sexual growth dimorphism in the half-smooth tongue sole (Cynoglossus semilaevis). We found that the full-length GHRH and PACAP gene sequences obtained from females and males consist of 4160, 4159, 2425, and 2446 bp, respectively, each of which includes four exons and three introns. When we analyzed normal females and males and extra-large male adults, GHRH and PACAP mRNA were found to be predominantly expressed in the brain; the expression levels were highest in normal males. The extra-large males exhibited the lowest mRNA levels of both GHRH and PACAP. Sex differences in GHRH and PACAP mRNA expression during development were also examined in a full-sib family; GHRH and PACAP mRNA were detected at all 27 times sampled from 10 to 410 days old. The GHRH expression levels in females were significantly higher than in males at most of the stages between 20 and 100 days old, while lower than those of males after 120 days old. Five microsatellite loci were identified in GHRH and PACAP genes. We used these five polymorphic markers to genotype 224 individuals, and no significant differences were found between females and males from the Bohai Sea, the Yellow Sea and hatchery samples.

The feasibility of using magnetic nanoparticles modified as gene vector / Viabilidad del uso de nanopartículas magnéticas modificadas como vectores genéticos

OBJECTIVE: To evaluate the feasibility of using magnetic nanoparticles (MNPs) as gene vector and the effect of magnetic field on efficiency of transfection. METHODS: Magnetic nanoparticles were prepared by controlling some chemical reaction parameters through a partially reduction precipitation method with ferric chloride aqueous solution as precursor material. The surface of particles was modified by polyethyleneimine (PEI) agents. The appearance, the size distribution, structure and phase constitute of MNPs were characterized by Transmission electron microscope (TEM), X-ray diffraction (XRD); the potential of absorbing DNA of MNPs was analysed by electrophoresis. Transfection was determined by delivering reporter gene, PGL2-control encoding luciferase, to different cell lines using MNPs-PLL as vector. The effect of magnetic field on the efficiency of transfection was determined using Nd-Fe-B permanent magnet. RESULTS: Foreign gene could be delivered to various cell lines by MNPs-PLL and expressed with high efficiency but the transfection efficiency and time course varied in the different cell lines studied. Magnetic field could enhance the efficiency of transfection by 5-10 fold. CONCLUSION: MNPs- PLL can be used as a novel non-viral gene vector in vitro, which offers a basis for gene delivery in vivo.

OBJETIVO: Evaluar la viabilidad del uso de nanopartículas magnéticas (MNPs) como vectores genéticos y el efecto de campo magnético en la eficiencia de la transfección. MÉTODOS: Se prepararon nanopartículas magnéticas mediante el control de algunos parámetros de la reacción química a través de un método de precipitación de reducción parcial con soluciones acuosas de cloruro férrico como el material precursor. La superficie de las partículas fue modificada mediante agentes de polietileneimina (PEI). La apariencia, el tamaño, distribución, estructura y constitución de fase de las MNPs, se caracterizaron mediante el microscopio electrónico de transmisión (MET), difracción de rayos X (DRX); el potencial de adsorber ADN de las MNPs se analizó mediante electroforesis; la transfección se determinó mediante el suministro del gene reportador de la luciferasa control PGL2, a diferentes líneas celulares usando MNPs - PLL como vectores. El efecto de campo magnético sobre la eficacia de la transfección se determinó usando el imán permanente NdFeB. RESULTADOS: El gene foráneo pudo suministrarse a varias líneas celulares mediante MNPs - PLL y expresarse con alta eficiencia pero la eficiencia de la transfección y el curso de tiempo variaron en las diferentes líneas celulares estudiadas. El campo magnético pudo mejorar la eficiencia de la transfección en 5-10 veces. CONCLUSION: Las MNPs - PLL pueden usarse como un nuevo vector genético no viral in vito, lo cual ofrece una base para el suministro del gene in vivo.

The feasibility of using magnetic nanoparticles modified as gene vector.

OBJECTIVE: To evaluate the feasibility of using magnetic nanoparticles (MNPs) as gene vector and the effect of magnetic field on efficiency of transfection. METHODS: Magnetic nanoparticles were prepared by controlling some chemical reaction parameters through a partially reduction precipitation method with ferric chloride aqueous solution as precursor material. The surface of particles was modified by polyethyleneimine (PEI) agents. The appearance, the size distribution, structure and phase constitute of MNPs were characterized by Transmission electron microscope (TEM), X-ray diffraction (XRD); the potential of absorbing DNA of MNPs was analysed by electrophoresis. Transfection was determined by delivering reporter gene, PGL2-control encoding luciferase, to different cell lines using MNPs-PLL as vector. The effect of magnetic field on the efficiency of transfection was determined using Nd-Fe-B permanent magnet. RESULTS: Foreign gene could be delivered to various cell lines by MNPs-PLL and expressed with high efficiency but the transfection efficiency and time course varied in the different cell lines studied. Magnetic field could enhance the efficiency of transfection by 5-10 fold. CONCLUSION: MNPs- PLL can be used as a novel non-viral gene vector in vitro, which offers a basis for gene delivery in vivo.