CDKN1C and NAP1L4 in human CDKN1C/KCNQ1OT1 imprinted domain are two key candidate genes responsible for BWS (Beckwith-Wiedemann syndrome) and cancer. In order to increase understanding of these genes in pigs, their cDNAs are characterized in this paper. By the IMpRH panel, porcine CDKN1C and NAP1L4 genes were assigned to porcine chromosome 2, closely linked with IMpRH06175 and with LOD of 15.78 and 17.94, respectively. By real-time quantitative RT-PCR and polymorphism-based method, tissue and allelic expression of both genes were determined using F1 pigs of Rongchang and Landrace reciprocal crosses. The transcription levels of porcine CDKN1C and NAP1L4 were significantly higher in placenta than in other neonatal tissues (P < 0.01) although both genes showed the highest expression levels in the lung and kidney of one-month pigs (P < 0.01). Imprinting analysis demonstrated that in pigs, CDKN1C was maternally expressed in neonatal heart, tongue, bladder, ovary, spleen, liver, skeletal muscle, stomach, small intestine, and placenta and biallelically expressed in lung and kidney, while NAP1L4 was biallelically expressed in the 12 neonatal tissues examined. It is concluded that imprinting of CDKN1C is conservative in mammals but has tissue specificity in pigs, and imprinting of NAP1L4 is controversial in mammalian species.

Ionizing radiation has long been known to induce heritable mutagenic change in DNA sequence. However, the genome-wide effect of radiation is not well understood. Here we report the molecular properties and frequency of mutations in phenotypically selected mutant lines isolated following exposure of the genetic model flowering plant Arabidopsis thaliana to fast neutrons (FNs). Previous studies suggested that FNs predominantly induce deletions longer than a kilobase in A. thaliana. However, we found a higher frequency of single base substitution than deletion mutations. Whilst the overall frequency and molecular spectrum of fast-neutron (FN)-induced single base substitutions differed substantially from those of "background" mutations arising spontaneously in laboratory-grown plants, G:C>A:T transitions were favored in both. We found that FN-induced G:C>A:T transitions were concentrated at pyrimidine dinucleotide sites, suggesting that FNs promote the formation of mutational covalent linkages between adjacent pyrimidine residues. In addition, we found that FNs induced more single base than large deletions, and that these single base deletions were possibly caused by replication slippage. Our observations provide an initial picture of the genome-wide molecular profile of mutations induced in A. thaliana by FN irradiation, and are particularly informative of the nature and extent of genome-wide mutation in lines selected on the basis of mutant phenotypes from FN-mutagenized A. thaliana populations.

BACKGROUND: Prohibitin (PHB) is an evolutionarily conserved multifunctional protein with ubiquitous expression. However, its molecular roles are largely unknown. METHODS: To better understand the function of prohibitin protein in silkworm (BmPHB), its coding sequence was isolated from a cDNA library of silkworm pupae. An His-tagged BmPHB fusion protein was expressed in Escherichia coli Rosetta (DE3) and purified with affinity and reversed-phase chromatography. Purified rBmPHB was used to generate anti-BmPHB polyclonal antibody. The subcellular localization of BmPHB was analysed by immunohistochemistry. RESULTS: BmPHB gene has an ORF of 825bp, encoding a predicted peptide with 274 amino acid residues. Immunostaining indicate that prohibitin is expressed in nucleus and predominately in cytoplasm. Western blot analyses indicated that, in the fifth instar larva, BmPHB was expressed descendingly in gonad, malpighian tubule, trachea, fatty body, intestine, and head. However, no expression was detected in larva's silk gland and epidermis. In addition, BmPHB was expressed in the nascent egg, larva and pupa, but not in the moth. CONCLUSIONS: The expression of BmPHB gene presents differential characteristic in different stage and tissues. It may play important roles in the development of silkworm. GENERAL SIGNIFICANCE: Studies on prohibitin have been still restricted to a few specific insects and insect cell lines such as Drosophila, Acyrthosiphon pisum and mosquito cell lines, not yet in silkworm. This is a first characterization of prohibitin in silkworm, B. mori.

To investigate the potential use of n-alkanes (alkanes), long-chain alcohols (alcohols) and long-chain fatty acids (acids) for estimating the diet composition of sheep, in a feeding trial. A total of 18 sheep were assigned randomly to three different diets (diet A, diet B and diet C) containing up to eight herbage species (Leymus chinensis, Leymus dasystachys, Elymus sibiricum, Chenopodium album, Puccinellia chinampoensis, Medicago sativa, Saussurea sinuata and Bromus inermis). Faecal recoveries of alkanes, alcohols and acids were determined, and diet compositions were estimated using different combinations of alkanes, alcohols and acids. The faecal concentrations of individual alkanes, alcohols and acids were corrected using the mean recovery of the dietary treatment that the respective animal belonged to (diet recovery), or the mean recovery across all dietary treatments (general recovery). In general, diets did not affect the faecal recovery values for alkanes, alcohols and acids, and no difference in accuracy was found between diet composition estimates based on dietary recovery and general recovery. The accuracy of diet composition estimates declined as the number of dietary components increased from four to eight herbage species (P < 0.001). Better (P < 0.05) estimates of diet composition were obtained with the combinations of two or three marker types instead of alkanes alone. Moreover, results showed that excluding minor diet components from the calculations decreased (P < 0.05) the accuracy of diet composition estimates, whereas including extra non-grazed herbage species did not reduce (P > 0.05) the quality of diet composition estimates. These results confirmed the usefulness of alkanes, alcohols and acids as markers for determining complex diet composition of sheep. However, a negative impact on the accuracy of diet composition estimates, caused by missing minor diet components from the calculation of diet composition, could happen when plant wax markers are used to estimate the diet composition of free-ranging animals.

Abstract Background: Although leukotriene D4 (LTD4) was a potent bronchoconstrictor, little is known about airway responsiveness to LTD4 in asthmatics with different inflammation phenotypes. Objectives: To establish the methodology and investigate the distribution characters of airway responsiveness, diagnostic value and safety of LTD4 bronchial provocation test. Methods: LTD4 bronchial provocation tests were performed in 62 asthmatics and 21 normal controls. Airway responsiveness was assessed based on the cumulative dosage causing a 20% fall in FEV(1)(PD(20)FEV(1)-LTD4) and was expressed as (median, interquartile range). The fall in spirometric parameters was plotted showing the distribution characters. The diagnostic value was assessed using receiver operation characteristic (ROC) curve. All adverse events were recorded during the test. Results: Airway responsiveness to LTD4 was significantly higher in asthmatics (0.410 nmol, 0.808 nmol) as compared with normal controls (5.00 nmol, 0.00 nmol). The decrease in spirometric parameters varied after bronchoprovocation, which was negatively correlated with PD(20)FEV(1)-LTD4, among which FEV(1) had a maximal slope (r=-0.524, P=0.000). High diagnostic value [AUC: 0.914, 95%CI:(0.855, 0.974)] was revealed by ROC curve. The major adverse events were dyspnea (82.3%), chest tightness (72.6%), wheezing (32.3%) and coughing (25.8%) in asthmatics, which could overall be recovered within 15.0 minutes after inhalation of 200∼400 mcg salbutamol MDI. No severe adverse event was reported. Conclusion: The established procedure of LTD4 bronchial provocation test is effective in the diagnosis of asthma and is well tolerated. Future studies are necessary to provide more evidences in terms of safety and efficacy. This may be helpful upon further application in clinical practice.

This paper proposes a solid model based on four-dimensional trivariate B-spline for strain and stress analysis of ventricular myocardium. With a series of processing steps in the four-dimensional medical images, the feature points of ventricular inner and outer wall are obtained. A B-spline surface is then used to build the dynamic deformation model of the myocardial walls. With such a surface model, a hexahedron control mesh can be constructed by sweeping the cloud data, and the ventricular solid model is built by fitting the trivariate B-spline parameters. Based on these models, a method of isogeometric analysis can be applied to calculate the stress and strain continuously distributed in the ventricle. The model is represented smoothly in the cylindrical coordinate system and is easy to measure myocardium dynamics for finding abnormal motion. Experiments are carried out for comparing the stress and strain distribution. It is found that the solid model can determine ventricular dynamics which can well reflect the deformation distribution in the heart and imply early clues of cardiac diseases.

We identified a novel gene encoding a Bombyx mori thymosin (BmTHY) protein from a cDNA library of silkworm pupae, which has an open reading frame (ORF) of 399 bp encoding 132 amino acids. It was found by bioinformatics that BmTHY gene consisted of three exons and two introns and BmTHY was highly homologous to thymosin betas (Tβ). BmTHY has a conserved motif LKHTET with only one amino acid difference from LKKTET, which is involved in Tβ binding to actin. A His-tagged BmTHY fusion protein (rBmTHY) with a molecular weight of approximately 18.4 kDa was expressed and purified to homogeneity. The purified fusion protein was used to produce anti-rBmTHY polyclonal antibodies in a New Zealand rabbit. Subcellular localization revealed that BmTHY can be found in both Bm5 cell (a silkworm ovary cell line) nucleus and cytoplasm but is primarily located in the nucleus. Western blotting and real-time RT-PCR showed that during silkworm developmental stages, BmTHY expression levels are highest in moth, followed by instar larvae, and are lowest in pupa and egg. BmTHY mRNA was universally distributed in most of fifth-instar larvae tissues (except testis). However, BmTHY was expressed in the head, ovary and epidermis during the larvae stage. BmTHY formed complexes with actin monomer, inhibited actin polymerization and cross-linked to actin. All the results indicated BmTHY might be an actin-sequestering protein and participate in silkworm development.

BACKGROUND AND OBJECTIVES Chronic total occlusion of the left main coronary artery (LMCA) is a rare condition, and the information on surgical experiences is limited. Although total occlusion of the LMCA is accompanied by well-developed collateral circulation, the condition of circulation is unstable during manipulation of the heart. We report our experience with revascularization in cases with total occlusion of the LMCA using the on-pump beating-heart (OnP-BH) technique. DESIGN AND SETTING Retrospective case review of patients treated at The First Affiliated Hospital of China Medical University over a 10-year period (1999 to 2009). PATIENTS AND METHODS The on-pump coronary artery bypass grafting with the beating heart was applied to 8 patients with chronic total occlusion of the LMCA. The extracorporeal circulation period, intubation duration, intensive care unit stay period, discharge period, preoperative and postoperative treatments, and follow-up were observed. RESULTS The mean extracorporeal circulation period was 80.4 (19.7) minutes. The mean intubation duration was 13.0 (4.6) hours. The mean intensive care unit stay period was 3.2 (0.7) days, and the mean discharge period was 16.8 (3.3) days. No perioperative myocardial infarction occurred. The mean follow-up period was 50.9 (34.8) months. All patients were asymptomatic, and no deaths were recorded during the follow-up period. The results of echocardiography showed improvement in the left ventricular function. CONCLUSIONS The OnP-BH myocardial revascularization seems to be a valid alternative for chronic total occlusion of the LMCA.

Crosslinking of ligand-engaged cytotoxic T lymphocyte antigen-4 (CTLA-4) to the T cell receptor (TCR) during the early phase of T cell activation attenuates TCR signaling, leading to T cell inhibition. To promote this event, a bispecific fusion protein comprising a mutant mouse CD80 (CD80w88a) and lymphocyte activation antigen-3 was engineered to concurrently engage CTLA-4 and crosslink it to the TCR. Crosslinking is expected to be attained via ligation of CTLA-4 first to MHC II and then indirectly to the TCR, generating a CTLA-4-MHC II-TCR tri-molecular complex that forms between T cells and antigen-presenting cells during T cell activation. Treating T cells with this bispecific fusion protein inhibited T cell activation. In addition, it induced the production of IL-10 and TGF-β, and attenuated AKT and mTOR signaling. Intriguingly, treatment with the bispecific fusion protein also directed early T cell differentiation into Foxp3 positive regulatory T cells (Tregs). This process was dependent on the endogenous production of TGF-β. Thus, bispecific fusion proteins that engage CTLA-4, and co-ligate it to the TCR during the early phase of T cell activation can negatively regulate the T cell response. Bispecific biologics with such dual functions may therefore represent a novel class of therapeutics for immune modulation. These findings presented here also reveal a potential new role for CTLA-4 in Treg differentiation.

Arc discharge of a hot cathode bucket ion source tends to be unstable what attributes to the filament self-heating and energetic electrons backstreaming from the accelerator. A regulation method, which based on the ion density measurement by a Langmuir probe, is employed for stable arc discharge operation and long pulse ion beam generation. Long pulse arc discharge of 100 s is obtained based on this regulation method of arc power. It establishes a foundation for the long pulse arc discharge of a megawatt ion source, which will be utilized a high power neutral beam injection device.