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It is challenging for radiologists and gynecologists to identify the type of ovarian lesions by reading magnetic resonance (MR) images. Recently developed convolutional neural networks (CNNs) have made great progress in computer vision, but their architectures still need modification if they are used in processing medical images. This study aims to improve the feature extraction capability of CNNs, thus promoting the diagnostic performance in discriminating between benign and malignant ovarian lesions.We introduce a feature fusion architecture and insert the attention models in the neural network. The features extracted from different middle layers are integrated with reoptimized spatial and channel weights. We add a loss function to constrain the additional probability vector generated from the integrated features, thus guiding the middle layers to emphasize useful information. We analyzed 159 lesions imaged by dynamic contrast-enhanced MR imaging (DCE-MRI), including 73 benign lesions and 86 malignant lesions. Senior radiologists selected and labeled the tumor regions based on the pathology reports. Then, the tumor regions were cropped into 7494 nonoverlapping image patches for training and testing. The type of a single tumor was determined by the average probability scores of the image patches belonging to it.We implemented fivefold cross-validation to characterize our proposed method, and the distribution of performance matrics was reported. For all the test image patches, the average accuracy of our method is 70.5% with an average area under the curve (AUC) of 0.785, while the baseline is 69.4% and 0.773, and for the diagnosis of single tumors, our model achieved an average accuracy of 82.4% and average AUC of 0.916, which were better than the baseline (81.8% and 0.899). Moreover, we evaluated the performance of our proposed method utilizing different CNN backbones and different attention mechanisms.The texture features extracted from different middle layers are crucial for ovarian lesion diagnosis. Our proposed method can enhance the feature extraction capabilities of different layers of the network, thereby improving diagnostic performance.

Particulate matter (PM) pollution in high emission regions will affect air quality, human health and climate change on both local and regional scales, and thus attract worldwide attention. In this study, a comprehensive study on PM

A novel strategy to fabricate bilayer hydrogel actuators based on the asymmetric distribution of crystalline regions across the bilayer structures was proposed. By employing PVA polymer chains into an alkali solvent-derived chitosan hydrogel matrix, chitosan/PVA hybrid bilayer hydrogels with both excellent responsive bending and mechanical properties were obtained as pH-controlled manipulators. In the design, the chitosan/PVA hydrogels upon treatment with freeze-thawing cycles were taken as the first monolayer, where excessive crystalline regions appeared. The original chitosan/PVA hydrogel as the second monolayer was then integrated into one bilayer device through the chemical-crosslinking of epichlorohydrin at the interface. The results showed that the resultant chitosan/PVA bilayer hydrogel actuator with a weight ratio of 3 : 1 displayed better sensitivity upon exposure to stimuli. The actuation behaviors are strongly dependent on experimental parameters such as the pH, PVA content and the chemical-crosslinking density. It is proposed that the driving force originates from the asymmetric distribution of crystalline regions, thus resulting in differential swelling ratios between the monolayers. In addition, programmable 3D shape transformations were achieved by using the bilayer hydrogel with designed 2D geometric patterns, and the tailored gripper-like hydrogel actuator can successfully capture and transport the cargo. Moreover, this actuation behavior can be erased and re-written on demand under certain conditions. Taking advantage of this universal strategy, more attractive actuators derived from synthetic or natural polymers in combination with PVA are highly expected, which can be used as smart soft robots in various fields such as manipulators, grippers, and cantilever sensors.

To investigate the relationship between serum superoxide dismutase (SOD) activity and the presence of chronic complications in patients with type 2 diabetes mellitus (T2DM). We conducted a retrospective cross-sectional study in patients with T2DM. They were assigned to three groups (Q1, Q2, and Q3) by SOD levels in both sexes. Clinical characteristics, cardiovascular disease, diabetic retinopathy, nephropathy, and peripheral neuropathy were compared. The relationship between the SOD and the prevalence of chronic complications was analyzed by binary logistic regression. Statistical analysis was performed in SPSS 26.0 (SPSS Inc., Chicago, IL). A total of 645 T2DM patients (401 men and 244 women) with complete data for SOD and medical records of complications were included. In men, patients in the Q1 group (lowest serum SOD activity) had the highest prevalence of diabetes with atherosclerosis (AS) (p

Microstructure of starch plays a vital role in the physicochemical properties. In this study, the microstructure, gelatinization and pasting properties of rice starches after acid, heat and acid-heat treatments were investigated. In our findings, molecular weight and size, amylopectin chain length distribution, degree of branching, amylose content, relative crystallinity, and hot water-solubility of rice starch were varied significantly after acid-heat treatment, showing a synergy between acid and heat. Correspondingly, acid-heat treatment markedly changed gelatinization and pasting properties of rice starch by sharply decreasing gelatinization and pasting temperatures as well as pasting viscosities. Pearson correlation analysis was performed, showing that molecular weight, size, amylopectin chain length distribution, and amylose content were significantly correlated to gelatinization temperatures, and crystallinity was remarkably correlated to gelatinization enthalpy. As for pasting properties, molecular weight and size as the major factors were substantially correlated to pasting viscosities and pasting temperature. This study provides a structure-property correlation for acid-heat treated rice starch, giving scientific basis for the application of starch in food industry.

Background Haemophilus parasuis is a commensal pathogen in the swine upper respiratory tract and causes Glässer’s disease. Surveillance, screening for infection, and vaccination response of H. parasuis is hindered by the lack of a rapid antibody detection method. Results In the present study, a monomeric autotransporter was identified as a novel antigen for developing an indirect ELISA. The autotransporter passenger domain (Apd) was expressed, purified, and demonstrated to be specific in ELISA and western blotting. Mouse antiserum of recombinant Apd (rApd) recognized native Apd in the 15 serotype reference strains and five non-typeable isolate stains, but showed no reaction with seven other bacterial pathogens. The rApd ELISA was optimized and validated using 67 serum samples with known background, including 27 positive sera from experimentally infected and vaccinated pigs along with 40 negative sera that had been screened with H. parasuis whole cell ELISA from clinically healthy herds. The rApd ELISA provided positive and negative percent agreements of 96.4 and 94.9%, respectively, and an AUC value of 0.961, indicating that the assay produced accurate results. Conclusion Apd was a universal antigen component among 15 serotype and non-typeable strains of H. parasuis and was also specific to this pathogen. The rApd ELISA could detect antibodies elicited by H. parasuis infection and vaccination, thereby exhibiting the potential to be applied for Glässer’s disease diagnosis, H. parasuis vaccination evaluation, and large-scale serological surveillance.

Glaesserella parasuis is an important bacterial pathogen that affects the swine industry worldwide. Research on the pathogenic mechanism and genetically engineered vaccine remains undeveloped because an effective markerless and multiple-gene knockout system is unavailable for G. parasuis yet. To establish a markerless knockout, deleted allelic genes with kanamycin resistance (Kan

In order to achieve the targets specified in the Action Plan for Air Pollution Prevention and Control (APAPPC), a limited coal banning area (10,000 km

Size controllable silver nanoparticles (AgNPs) were synthesized in situ on the polysaccharides-based nanotubes, which were formed by the triple-helix polysaccharide extracted from black fungus (AF1). The results of transmission electron microscope (TEM), scanning electron microscope (SEM) and energy dispersive spectrometer (EDS) proved that AgNPs with the size from 10-25 nm were uniformly dispersed on the surface of AF1 dendritic nanotubes without affecting their tubular morphology. Moreover, due to the tubular structure, the loaded silver content of the composites (AgNPs and AF1 nanotube, AF1-Ag) could reach about 50% by thermogravimetric analysis (TG) evaluation. Thus, the smaller size of AgNPs and higher silver loading content suggest that the composites could be applied in the biomedical field. The antibacterial properties of AF1-Ag were evaluated as an example in the present work. As expected, the culture medium contained a few of AF1-Ag (10% ω%, c = 50 μg/mL) exhibited obvious antibacterial properties, and the effect of bacteriostasis increased with the increase of the amount of supported silver content. Taken together, the AF1-Ag with good antibacterial activity and good stability has the potential to be applied in the antibacterial field.

Natural polysaccharides as sustainable polymers are rich sources with good biological safety and various biological functions, which are important research topics in the fields of food and medicine. However, their widespread medical application is hindered due to the technical difficulties in the extraction/purification and chain structure characterization of polysaccharides, as well as due to the lack of thorough understanding of the macromolecular chain conformation and the mechanism of bioaction for these polysaccharides. To provide a relatively comprehensive reference for relevant researchers, we mainly introduce several natural polysaccharides with different chain conformations including random coil, hyperbranched sphere, and triple helix in solution, as well as summarize their extraction and purification processes, characterization of chemical structure and chain conformation, anti-tumor activities and potential mechanisms.

Ulcerative colitis (UC) is a complex disease that results from a dysregulated immune response in the gastrointestinal tract. A mouse model orally administered with dextran sodium sulfate (DSS) is the most widely used experimental animal model of UC. However, the ultrastructure of the colon in mouse colitis is poorly understood. In the present study, colonic specimens from DSS-induced UC mice underwent hematoxylin and eosin staining, Masson's trichrome staining and Verhoeff's elastic staining. In addition, the ultrastructure of samples was examined by transmission electron microscopy. UC was successfully induced by 7 consecutive days of DSS oral administration. The goblet cell architecture of the UC tissue was damaged in the mucosa. Additionally, a significant number of inflammatory cells infiltrated into the stroma and the structure of the intestinal gland was destroyed. The tissue in the submucosa showed significant edema. Hyperplasia was also identified in the submucosa, promoting a disorganized microstructure within the colon wall. Numerous collagen fibers in the muscular layer were disrupted, and the fiber bundles were thinner compared with those in the normal control group. Furthermore, in the DSS-induced UC group, the smooth muscle cell showed edema, the cell membrane structure was unclear and the shape of the nucleus was irregular. In conclusion, the present study revealed important histological and ultrastructural changes in the colon of DSS-induced UC mice. These features may contribute to improved understanding of the pathogenesis and mechanism of UC.

The water soluble β-(1 → 3)-D-glucan with short branches (AF1) isolated from Auricularia auricula-judae was successfully fractionated by ultrasonication into three fractions with different weight-average molecular weights (Mws). The results of static and dynamic laser light scattering, viscometry and atomic force microscopy confirmed that the AF1 samples adopted a stiff chain conformation in water, and the coexistence of individuals and aggregates occurred gradually with increasing concentration. The AF1 sample with the highest Mw easily self-entangled, and exhibited a strong shear rate-dependence of viscosity in water. The glucans displayed anti-hepatoma activity and significantly inhibited H22 tumour growth without cytotoxicity towards normal tissues. They displayed both molecular weight- and dosage-dependencies of anti-tumour activity, and the sample with an Mw of 7.7 × 105 at the dosage of 5 mg kg−1 exhibited the highest inhibition ratio of ∼77% against H22 tumour, even significantly higher than the positive control of cytoxan. The immunohistochemical and western blot analyses revealed that the AF1 glucans triggered cell apoptosis, indicated by the activation of caspase 3/9 and down-regulated tumour angiogenesis factors of VEGF and CD31. The underlying antitumor mechanism was suggested to induce tumour cell apoptosis and to inhibit angiogenesis in tumour tissues via enhancement of the immune-response. Taken together, the AF1 β-glucan was a potent natural drug candidate with high anti-cancer activities and less cytotoxicity, and the AF1 sample with a moderate molecular weight existed in aqueous solution as a more extended chain conformation, which plays an important role in activating immune responses.

A novel gene carrier was prepared from a triple-helical β-glucan LNT (t-LNT) on the basis of the special interaction between the single chain of t-LNT (s-LNT) and polydeoxyadenylic acid (poly(dA)). The effects of the chain length of poly(dA) in the target DNA sequence on the stability and gene transfection efficiency of the gene carrier of the poly(dA)–s-LNT complex were evaluated by agarose gel retardation assay, circular dichroism, flow cytometry, and ELISA kits. All the results demonstrated that the chain length of poly(dA) that produced the strongest interaction with s-LNT was favorable for the stability and gene transfection efficiency. Moreover, the gene transfection efficiency was strikingly dependent on the transfection time and temperature, suggesting the internalization of the gene delivery system via endocytosis. The high expressions of receptors including TLR2 and CR3 (CD11b/CD18) on the surfaces of RAW264.7 cells were confirmed by confocal microscopy and flow cytometry. By blocking TLR2, CD11b, and CD18 with the corresponding specific antibodies, the transfection efficiency was hardly changed at 4 °C or 37 °C. This suggested that the internalization pattern of such a delivery system was not via the usual β-glucan receptor-mediated endocytosis (such as dectin-1, TLR2, and CR3), which was different from the reported uptake mechanism. Overall, this work facilitates the understanding of the pathways of delivery systems involving β-glucans, which has significant implications for potential applications in disease treatment.

Excess bilirubin often evokes hepatobiliary system dysfunction. In the present work, we developed an efficient, safe and blood compatible adsorbent for bilirubin removal from human blood. In view of the highly effective adsorption of carbon nanotubes (CNTs) on bilirubin but with many side effects, and good biocompatibility of chitin but with low efficiency for bilirubin removal, new chitin/carbon nanotube (Ch/CNT) nanofibrous microspheres were constructed from chitin solution in NaOH/urea aqueous system by blending with CNTs. The results of AFM, SEM, and TEM demonstrated that the CNTs were dispersed well in the chitin matrix, and the chitin nanofibers intertwined with CNTs to form hybrid chitin/CNT nanofibers and then weaved into a 3D interconnected network architecture. Moreover, lysine (Lys), a highly specific ligand for bilirubin, was immobilized tightly to the hybrid microspheres to obtain Ch/CNT/Lys. The resultant microspheres possessed large surface area and hierarchical pores including mesopores and micropores, which could allow bilirubin to enter easily and store, leading to highly efficient adsorption. The Ch/CNT/Lys microspheres exhibited excellent bilirubin adsorption property (107.2 mg g−1) and efficient bilirubin clearance rate from real hyperbilirubinemia plasma competing with protein, as well as good cell affinity and blood compatibility, as a result of the combination of the high adsorption of CNTs and inherent biocompatibility of chitin and lysine. Therefore, an effective strategy to develop a novel biocompatible and blood compatible bilirubin adsorbent is provided, showing potential applications for hemoperfusion in blood purified therapy.

Dendritic nanotubes (DNTs) with hydrophobic cavities were constructed directly from rigid branched β-1,3-D-glucan (AF1) in aqueous solution, and the AF1 sample was isolated from the fruiting bodies of Auricularia auricula-judae, a household nutritional food. The structure of AF1 dendritic nanotubes was demonstrated with a transmission electron microscope (TEM) and a scanning electron microscope (SEM), and a schematic diagram was proposed to describe the formation process, which was supported by the results of static/dynamic light scattering (SLS/DLS) and atomic force microscopy (AFM). In solution, a sequential self-assembly of the AF1 chains in a parallel manner occurred to form lamellas followed by self-curling into nanotubes with the mean diameters from 20 to 80 nm, depending on the concentration and molecular weight of AF1, through hydrogen bonding and hydrophilic/hydrophobic interaction. As a result of the dendritic structure, the AF1 aggregates exhibited highly condensed hydrophobic regions, which could be used as carriers to achieve a high concentration of the target molecules. In our findings, the anticancer drug DOX and the fluorescent probe TPA-BMO could be loaded into the hydrophobic region of DNTs. Interestingly, DOX-loaded DNTs of AF1 exhibited high drug loading capacity and pH-triggered sustained release behaviors (>23 days) with reduced cytotoxicity in vitro. Moreover, the bioimaging experiment demonstrated that TPA-BMO-loaded DNTs of AF1 induced stronger fluorescence intensity than TPA-BMO alone, and maintained a longer duration time (18 days) in vivo. Therefore, the DNTs of AF1 have promising applications as bioactive carriers, especially in the fields of drug delivery and bioimaging.

A green method for in situ one-pot synthesis of size-controlled gold nanoparticles (AuNPs) was provided in this work. In our findings, a β-1,3-D-glucan, AF1, extracted from black fungus was used as a support of AuNPs. The AuNPs were dispersed in the AF1 aqueous solution without additional reducing and stabilizing or dispersing agents. The infrared spectrum (FT-IR), thermogravimetry (TG), X-ray photoelectron spectra (XPS) and atomic force microscope (AFM) results indicated that the AuNPs synthesized by AF1 were immobilized on the AF1 nanotubes with controllable shape and size of 5-45nm. The hydroxyl groups on AF1 nanotubes not only acted as reducing agents but also were able to stabilize AuNPs by the strong interaction between the surface Au atoms of AuNPs and oxygen atoms of AF1 nanotubes. The AuNPs exhibited a good catalytic activity for the rapid reduction from 4-nitrophenol (4-NP) to 4-aminophenol (4-AP) by excess borohydride, further confirming the dispersion of AuNPs in water.

Increasing the level of reactive oxygen species (ROS) in cancer cells has been suggested as a viable approach to cancer therapy. Our previous study has demonstrated that mitochondria-targeted flavone-naphthalimide-polyamine conjugate 6c elevates the level of ROS in cancer cells. However, the detailed role of ROS in 6c-treated cancer cells is not clearly stated. The biological effects and in-depth mechanisms of 6c in cancer cells need to be further investigated. In this study, we confirmed that mitochondria are the main source of 6c-induced ROS, as demonstrated by an increase in 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA) and MitoSox fluorescence. Compound 6c-induced mitochondrial ROS caused mitochondrial dysfunction and lysosomal destabilization confirmed by absolute quantitation (iTRAQ)-based comparative proteomics. Compound 6c-induced metabolic pathway dysfunction and lysosomal destabilization was attenuated by N-acetyl-L-cysteine (NAC). iTRAQ-based comparative proteomics showed that ROS regulated the expression of 6c-mediated proteins, and treatment with 6c promoted the formation of autophagosomes depending on ROS. Compound 6c-induced DNA damage was characterized by comet assay, p53 phosphorylation, and γH2A.X, which was diminished by pretreatment with NAC. Compound 6c-induced cell death was partially reversed by 3-methyladenine (3-MA), bafilomycin (BAF) A1, and NAC, respectively. Taken together, the data obtained in our study highlighted the involvement of mitochondrial ROS in 6c-induced autophagic cell death, mitochondrial and lysosomal dysfunction, and DNA damage.

The aim of the current study was to investigate the effects of early-stage dextran sodium sulfate (DSS)-induced mouse colitis on the biomechanical properties and microstructure of colon walls. In the present study, colitis was induced in 8-week-old mice by the oral administration of DSS, and then 10 control and 10 experimental colitis samples were harvested. Uniaxial tensile tests were performed to measure the ultimate tensile strength and ultimate stretches of colon tissues. In addition, histological investigations were performed to characterize changes in the microstructure of the colon wall following treatment. The results revealed that the ultimate tensile stresses were 232±33 and 183±25 kPa for the control and DSS groups, respectively (P=0.001). Ultimate stretches at rupture for the control and DSS groups were 1.43±0.04 and 1.51±0.06, respectively (P=0.006). However, there was no statistically significant difference in tissue stiffness between the two groups. Histological analysis demonstrated high numbers of inflammatory cells infiltrated into the stroma in the DSS group, leading to significant submucosa edema. Hyperplasia was also identified in the DSS-treated submucosa, causing a disorganized microstructure within the colon wall. Furthermore, a large number of collagen fibers in the DSS-treated muscular layer were disrupted, and fiber bundles were thinner when compared with the control group. In conclusion, early-stage experimental colitis alters the mechanical properties and microstructural characteristics of the colon walls, further contributing to tissue remodeling in the pathological process.

Polysaccharides are naturally occurring biological macromolecules that are envisaged as promising substitutes of non-degradable polymers due to their outstanding inherent properties of biodegradability, biocompatibility, low-cost, and availability. Their utilization in the development of nanostructured hybrid materials has numerous advantages in theranostics, the integrated approach of therapeutics and diagnostics. In particular, some rigid polysaccharides occur in nature, which can self-assemble into ordered hierarchical structures, facilitating the controllable fabrication of various nanocomposites. These rigid polysaccharides, including Lentinan, Curdlan, Schizophyllan, Scleroglucan, Auricularian, and yeast glucan, possess the linear β-(1→3)-d-glucan as the backbone with or without β-(1→6)-linked glucose as the branch, having diverse biological activities. This review focuses on the incorporation of nanoparticles, such as gold (AuNPs), silver (AgNPs), selenium (SeNPs), silica, carbon nanotubes, homo-polynucleotides, bio-imaging probes, and drugs, into different rigid polysaccharide matrices through self-assembly and summarizes their biological functions as well as the correlation to their conformations. Additionally, we addressed the use of the as-engineered polysaccharide nanocomposites as effective therapeutic agents, and the challenges or ambiguity issues concerning further practical clinic therapeutic applications.

Quinoa flour (QF) was added in wheat flour to make nutritionally fortified wheat bread (WB). Effects of QF on baking performance, antioxidant activity and digestibility of WB were studied. The results indicated that with an addition of a small amount of QF (5%) would not affect the baking performance of WB, while higher amounts of QF (10% and 15%) resulted in smaller specific volume, increased hardness and coarse porosity, since QF changed gluten secondary structure and disrupted gluten network. However, sensory evaluation showed that the flavor of WB enhanced significantly with increasing QF addition. More importantly, WB containing QF exhibited improved antioxidant activity and reduced in vitro digestibility with lower estimated glycemic index (eGI) and higher content of slowly digestible starch (SDS) and resistant starch (RS). Our findings indicate that QF-fortified bread is promising to be developed as a functional food with relatively lower eGI and more effective antioxidant properties.

Catecholaminergic polymorphic ventricular tachycardia (CPVT) is caused by a single point mutation in the cardiac type 2 ryanodine receptor (RyR2). Using a knockin (KI) mouse model (R2474S/+), we previously reported that a single point mutation within the RyR2 sensitizes the channel to agonists, primarily mediated by defective interdomain interaction within the RyR2 and subsequent dissociation of calmodulin (CaM) from the RyR2. Here, we examined whether CPVT can be genetically rescued by enhancing the binding affinity of CaM to the RyR2. We first determined whether there is a possible amino acid substitution within the CaM-binding domain in the RyR2 (3584–3603 residues) that can enhance its binding affinity to CaM and found that V3599K substitution showed the highest binding affinity of CaM to the CaM-binding domain. Hence, we generated a heterozygous KI mouse model (V3599K/+) with a single amino acid substitution in the CaM-binding domain of the RyR2 and crossbred it with the heterozygous CPVT-associated R2474S/+-KI mouse to obtain a double-heterozygous R2474S/V3599K-KI mouse model. The CPVT phenotypes — bidirectional or polymorphic ventricular tachycardia, spontaneous Ca(2+) transients, and Ca(2+) sparks — were all inhibited in the R2474S/V3599K mice. Thus, enhancement of the CaM-binding affinity of the RyR2 is essential to prevent CPVT-associated arrhythmogenesis.

Fabry disease (FD) was an X-linked lysosomal storage disorder resulting from a deficiency in glycosphingolipid catabolism caused by mutations in the α-galactosidase A gene GLA. Variant FD patients did not present with classical symptoms during childhood and were undiagnosed or misdiagnosed with other kidney diseases, such as chronic glomerulonephritis (CGN). In this study, we utilized exome sequencing and Sanger sequencing identified the variation p.E66Q of GLA completely co-segregated with the disease phenotype in a Chinese family, which previously been diagnosed as possible CGN. Female patients exhibited preferential X-chromosome inactivation (XCI) of the normal p.E66 allele, as indicated by XCI analysis. By measuring α-Gal A activity, we found that male patients in the pedigree had just little enzymatic activity while female patients had residual enzymatic activity. These patients were diagnosed with renal variant FD in subsequent clinical review. Our results directly implicated the GLA mutation p.E66Q as the genetic etiology of the Chinese renal variant FD pedigree.

Haemophilus parasuis is the etiological agent of Glässer's disease which is characterized by fibrinous polyserositis, arthritis and meningitis. The pathogenesis of this bacterium remains largely unknown. Genes expressed in vivo may play an important role in the pathogenicity of H. parasuis. The development of in vivo-induced antigen technology (IVIAT) has provided a valuable tool for the identification of in vivo-induced genes during bacterial infection. In this study, IVIAT was applied to identify in vivo-induced antigens of H. parasuis. Pooled swine H. parasuis-positive sera, adsorbed against in vitro-grown cultures of H. parasuis SH0165 and Escherichia coli BL21 (DE3), were used to screen the inducible expression library of genomic proteins from whole genome sequenced H. parsuis SH0165. Finally, 24 unique genes expressed in vivo were successfully identified after secondary and tertiary screening with IVIAT. These genes were implicated in cell surface proteins, metabolism, stress response, regulation, transportation and other processes. Quantitative real-time PCR showed that the mRNA levels of 24 genes were all upregulated in vivo relative to in vitro, with 13 genes were detected significantly upregulated in H. parasuis infected pigs. Several potential virulence-associated genes were found to be uniquely expressed in vivo, including espP, lnt, hutZ, mreC, vtaA, pilB, tex, sunT and aidA. The results indicated that the proteins identified using IVIAT may play important roles in the pathogenesis of H. parasuis infection in vivo.

The present study assessed the variation of measles viruses (MV) and its association with clinical manifestations in patients with MV. A total of 38 pediatric patients with MV at the acute infection stage were selected and 2 ml venous blood was collected from each of them. Serum immunoglobulin M antibodies were determined by ELISA. Urine specimens were collected from 30 of the 38 patients and associated genetic structures were detected by reverse-transcription polymerase chain reaction mapping. At the same time, clinical epidemiological manifestations were collected to perform an epidemiological analysis. The MV-positive rate within the cohort determined in serum was 100%. Seven MV strains were isolated from urine specimens of 30 patients and the positive rate was 23.33%. Four MV strains were randomly selected from the 7 strains and the results revealed that they were all of the H1a genotype. In addition, there was no significant correlation between clinical manifestation of pediatric patients with measles and the genotype of the MV. In conclusion, the preponderant genotype of MV in Kunming was H1a and there was obvious nucleotide or amino acid mutation. The clinical manifestation of MV infection in pediatric patients was not associated with the MV genotype.

Autism is a childhood neurodevelopmental disorder with high heterogeneity. Following our genome-wide associated loci with autism, we performed sequencing analysis of the coding regions, UTR and flanking splice junctions of AMPD1 in 830 Chinese autism individuals as well as 514 unrelated normal controls. Fourteen novel variants in the coding sequence were identified, including 11 missense variants and 3 synonymous mutations. Among these missense variants, 10 variants were absent in 514 control subjects, and conservative and functional prediction was carried out. Mitochondria activity and lactate dehydrogenase assay were performed in 5 patients' lymphoblast cell lines; p.P572S and p.S626C showed decreased mitochondrial complex I activity, and p.S626C increased lactate dehydrogenase release in medium. Conclusively, our data suggested that mutational variants in AMPD1 contribute to autism risk in Han Chinese population, uncovering the contribution of mutant protein to disease development that operates via mitochondria dysfunction and cell necrosis.

Autism is a neurodevelopmental disorder clini- cally characterized by impairment of social interaction, defi- cits in verbal communication, as well as stereotypic and repetitive behaviors. Several studies have implicated that abnormal synaptogenesis was involved in the incidence of autism. Neuroligins are postsynaptic cell adhesion molecules and interacted with neurexins to regulate the fine balance between excitation and inhibition of synapses. Recently, mutation analysis, cellular and mice models hinted neuroligin mutations probably affected synapse maturation and function. In this study, four missense variations (p.G426S (NLGN3), p.G84R (NLGN4X), p.Q162 K (NLGN4X) and p.A283T (NLGN4X)) in four different unrelated patients have been identified by PCR and direct sequencing. These four missense variations were absent in the 453 controls and have not been reported in 1000 Genomes Project. Bioinformatic analysis of the four missense variations revealed that p.G84R and p.A283T were ''Probably Damaging''. The variations may cause abnormal synaptic homeostasis and therefore trigger the patients more predisposed to autism. By case-control ana- lysis, we identified the common SNPs (rs3747333 and rs3747334) in the NLGN4X gene significantly associated with risk for autism (p = 5.09E-005; OR 4.685 (95 % CI 2.073-10.592)). Our data provided a further evidence for the involvement of NLGN3 and NLGN4X gene in the pathogen- esis of autism in Chinese population.

We have firstly demonstrated the renaturation process of dissociated single chains of lentinan (s-LNT) and the variable conformations of the renatured LNT (r-LNT). The results from ultrasensitive differential scanning calorimetry and circular dichroism revealed that the variable structures including perfect triple helix, defective triple helix containing duplex segment, and single chains occurred in the renaturation of s-LNT, depending on the renaturation time, solvent composition, molecular weight, and the mode of renaturation. When water was added into s-LNT/dimethylsulfoxide (DMSO) to reach 95% (v/v), the classic low-temperature intra-triple-helical conformational transition at ∼10°C (T1) appeared within 4 h, indicative of a rapid reconstruction of triple helical structure. Besides, one newly endothermic peak at ∼43°C (T2) simultaneously occurred, which was first ascribed to the melting of duplex segment in the imperfect triplex. The duplex stretches disappeared when DMSO reached 50%, in which single chains coexisted with triplex. Moreover, the duplex segment disappeared by slowly dropping water into s-LNT/DMSO. This work suggested that the structure of r-LNT could be controllable, and provided important information for their successful development and application in polymer and life science. © 2012 Wiley Periodicals, Inc. Biopolymers 97:988–997, 2012.

Autism is a heterogeneous childhood neurodevelopmental disorder that is characterised by deficits in verbal communication, impaired social interactions, restricted interests and repetitive behaviours. Using an Illumina HumanCNV370-Quad BeadChip, we identified two Han Chinese individuals with autism and large duplications (~1.6 Mb and ~2.4 Mb) disrupting the same CNTN4 gene. CNTN4 encodes a protein that functions as a cell-adhesion molecule and may play an essential role in the formation of axon connections in the developing nervous system. The disruption of this gene has been reported to be the cause of the 3p deletion syndrome and also a possible susceptibility factor for autism spectrum disorders (ASDs). Our results suggest that rare copy number variations (CNVs) in CNTN4 may also influence autism susceptibility in Asian populations. Interestingly, a comparison of the clinical phenotypes between the two subjects revealed that the subject with the 2.4 Mb CNV (involving several other genes) presented with a more severe phenotype than the subject with the 1.6 Mb CNV (disrupting only CNTN4 and CNTN6). This suggests that other genes in the nearby region may contribute to the pathogenesis.

Osteogenesis imperfect (OI) is a heritable connective tissue disorder with bone fragility as a cardinal manifestation, accompanied by short stature, dentinogenesis imperfecta, hyperlaxity of ligaments and skin, blue sclerae and hearing loss. Dominant form of OI is caused by mutations in the type I procollagen genes, COL1A1/A2. Here we identified a novel splicing mutation c.3207+1G>A (GenBank ID: JQ236861) in the COL1A1 gene that caused type I OI in a Chinese family. RNA splicing analysis proved that this mutation created a new splicing site at c.3200, and then led to frameshift. This result further enriched the mutation spectrum of type I procollagen genes.

The aim of the present study was to determine the antimicrobial susceptibility of Haemophilus parasuis in China and to investigate possible correlations between antimicrobial resistance and serovars or virulence. The susceptibility of 110 clinical isolates to 22 antimicrobial agents was determined by an agar dilution method. Except 28 isolates (25.5%) were non-typable, the isolates comprising thirteen serovars (1-5, 7-10, and 12-15), with the most prevalent were serovars 4 (14.5%), 5 (26.4%), 12 (9.1%) and 13 (10.9%). All isolates were susceptible to cefotaxime, ceftiofur, azithromycin, chloramphenicol, florfenicol and tilmicosin, and most of them were susceptible to penicillin, ampicillin, cefaclor, levofloxacin, ciprofloxacin, erythromycin, tetracycline, gentamicin, spectinomycin and tiamulin; however, a high level of resistance was encountered for enrofloxacin (70.9%) and trimethoprim/sulphamethoxazole (44.5%). There were 32 antimicrobial antibiograms observed, with 23.6% of the isolates resistant to more than 3 antimicrobial agents. Antimicrobial resistance was observed in every serovar included in this study, but no statistically significant association was observed between the antimicrobial resistance and the serovars or virulence of H. parasuis.

The gelation behavior of the triple-helical polysaccharide lentinan fractions having different molecular weights in water at 25 degrees C were studied by using a rheometer. The analysis of concentration and molecular weight dependence of shear stress and shear viscosity showed that aqueous lentinan is a typical shear-thinning fluid, possessing potential as a viscosity control agent, and that a weak gel with entangled network structure formed. The dynamic oscillatory behavior of lentinan in the temperature range of 1-15 degrees C was also investigated by rheologic method. The storage modulus G' and complex viscosity eta* increased first with decreasing temperature, and underwent a maximum centered at 7-9 degrees C, and then decreased with further decreasing temperature. This abnormal phenomenon was ascribed to formation of rigid structure in the gel state, which was confirmed by the experimental results from micro-DSC. The micro-DSC curves showed that an endothermic peak appeared at 7-9 degrees C for lentinan in water upon heating, which was attributable to the intramolecular order-disorder structure transition similar to triple-helical polysaccharide schizophyllan. Namely, at lower temperature, the side glucose residues of lentinan (triplix II) formed a well-organized triple-helical structure (triplix I) through hydrogen-bonding with the surrounding water molecules. Moreover, this conformation transition was proved to be thermally reversible.

Haemophilus parasuis is generally considered a commensal organism in the upper respiratory tract of pigs, where it evades the host immune system and survives the challenging host environment. In response to various host stresses, H. parasuis strains can adapt to the adverse conditions. However, the specific bacterial factors that participate in this process are poorly understood. Here, we investigated the role of ClpP in H. parasuis virulent strain CF7066 by generating a clpP deletion mutant (ΔclpP), as well as a complemented strain C-clpP. Our findings supported that ClpP is essential for stress tolerance of H. parasuis, by the demonstrations that the ΔclpP mutant showed decreased resistance to heat, oxidation, and osmotic pressure. Notably, we observed increased autoagglutination and biofilm formation in the ΔclpP mutant and the amount of polysaccharides and extracellular proteins, which are the main components of biofilm, were much higher in the ΔclpP mutant than the wild-type strain. Real-time PCR demonstrated that the transcriptional regulators csrA and rpoD, and a possible biofilm repressor luxS were significantly downregulated upon clpP deletion. Together, these observations suggest that ClpP plays an essential role in stress tolerance, and negatively regulates biofilm formation in H. parasuis.

Four water-insoluble (1--3)-alpha-D-glucans, coded L-II1, L-II2, L-II3 and L-II4, with different molecular weights were isolated from four kinds of fruiting bodies of Lentinus Edodes. The four alpha-D-glucans were O-sulfonated to obtain derivatives (SL-II) having degrees of substitution (DS) from 0.9 to 2.1 respectively. The structure of the samples was analyzed by infrared spectra, elemental analysis, and 13C NMR. The weight-average molecular weight (Mw), radii of gyration (s2z1/2) and intrinsic viscosity ([eta]) of the native alpha-D-glucans and O-sulfonated derivatives were measured by size-exclusion chromatography combined with laser light scattering (SEC-LLS), LLS, and viscometry in 0.2 M aqueous NaCl and in dimethyl sulfoxide (DMSO) containing 0.25 M LiCl at 25 degrees C respectively. The Mw values of the O-sulfonated derivatives were much lower than those of the native alpha-D-glucans. The experimental results indicate that the O-sulfonated derivatives are water-soluble and exist as an expanded flexible chain in aqueous solution owing to intramolecular hydrogen bonding or interaction between charge groups. The in vivo and in vitro antitumor activities of the native alpha-D-glucans and their O-sulfonated derivatives against solid tumor Sarcoma 180 cells were evaluated and compared. Interestingly, all of the O-sulfonated derivatives exhibited higher antitumor activities than those of the native glucans. The results reveal that the effect of O-sulfonation of the alpha-D-glucan on the improvement of their antitumor activities was considerable.

A water-soluble heteropolysaccharide ac-PCM0 from Poria cocos was successfully fractionated using a preparative size exclusion chromatography (SEC) column, and their weight-average molecular mass (M(w)) was characterized by analytical SEC combined with laser light scattering (SEC-LLS). The results indicate that the fractions having relatively high M(w) exhibited higher inhibition ratio in vivo antitumor activity than those having M(w) below 3.29 x 10(4). However, the relatively low molecular mass was beneficial to the in vitro antitumor activity. Moreover, ac-PCM0 has a significantly higher enhancement ratio of the body weight than 5-fluorouracil, and its 50% lethal dose is above 1250 mg/kg, indicating a nontoxic nature.

The present study investigated the functional involvement of the rat hippocampal formation in easy and difficult visual discrimination learning by measuring regional brain cytochrome oxidase (C.O.) activity, an index of neuronal metabolic activity. The results showed that learning was related to C.O. activity in the CA field and the dentate gyrus of the hippocampal formation of rats in the difficult discrimination, whereas learning was related to C.O. activity only in the dentate gyrus of rats in the easy discrimination. The results suggest that difficult visual discrimination learning required greater involvement of the hippocampal formation than easy visual discrimination learning.

Aeromonas (A) gum, an extracellular heteropolysaccharide produced by the bacterium Aeromonas nichidenii strain 5797, was studied by 1H and 13C NMR spectroscopy including 2D COSY, TOCSY, 1H, 13C HMQC, HMBC and ROESY experiments after O-deacetylation and Smith degradation. These investigations revealed the presence of an O-acetylated pentasaccharide repeating unit composed of mannose, glucose, xylose and glucuronic acid, and it has the following structure: [Image: see text]

Dynamic light scattering measurements have been made on 15 fractions of aeromonas (A) gum, an extracellular heteropolysaccharide produced by the strain Aeromonas nichidenii, with dimethylsulfoxide containing 0.2M lithium chloride as the solvent at 25°C. Data for the translational diffusion coefficient D covering a molecular weight range from 4.5 × 105 to 2.1 × 106 and ratios of the z-average radius of gyration 〈s2〉z1/2 to the hydrodynamic radius RH (calculated with previous 〈s2〉z data) suggest that the polymer behaves like a semiflexible chain in this solvent similar to the stiffness of cellulose derivatives. Thus the D data are analyzed on the basis of the Yamakawa–Fujii theory for the translational friction coefficient of a wormlike cylinder by coarse-graining the heteropolysaccharide molecule. Excluded-volume effects are taken into account in the quasi-two-parameter scheme, as was done previously for 〈s2〉z and [η] (the intrinsic viscosity) of A gum in the same solvent. The molecular weight dependence of RH is found to be explained by the perturbed wormlike chain with a persistence length of 10 nm, a linear mass density of 1350 nm−1, an excluded-volume strength parameter of 1.3 nm, and a chain diameter of 2.8 nm. These parameters are in substantial agreement with those estimated previously from 〈s2〉z and [η] data, demonstrating that the solution properties (D, 〈s2〉z, and [η]) of the heteropolysaccharide are almost quantitatively described by the current theories for wormlike chains in the molecular weight range studied. © 2002 Wiley Periodicals, Inc. Biopolymers 65: 387–394, 2002

To investigate the impact of Qing'e Pill (, QEP) on the cancellous bone microstructure and its effect on the level of β-catenin in a mouse model of postmenopausal osteoporosis.Ninety-six 8-week-old specific pathogen free C57BL/6 mice were randomly divided into 4 groups (24/group): sham, ovariectomised osteoporosis model, oestradiol-treated, and QEP-treated groups. Three months after surgery, the third lumbar vertebra and left femur of the animals were dissected and scanned using micro-computed tomography (micro-CT) to acquire three-dimensional (3D) parameters of their cancellous bone microstructure. The impact of ovariectomy, the effect of oestradiol and QEP intervention on cancellous bone microstructure, and the expression of β-catenin were evaluated.The oestradioland the QEP-treated groups exhibited a significant increase in the bone volume fraction, trabecular number, trabecular thickneßs, bone surface to bone volume ratio (BS/BV), and β-catenin expression compared with those of the model group (P0.05). In contrast, the structure model index, trabecular separation, and BS/BV were significantly decreased compared with those of the ovariectomised osteoporosis model group (P0.05). No differences were observed in the above parameters between animals of the QEP- and oestradiol-treated groups.The increased β-catenin expression may be the mechanism underlying QEP's improvement of the cancellous bone microstructure in ovariectomised mice. Our findings provide a scientific rationale for using QEP as a dietary supplement to prevent bone loss in postmenopausal women.

In colorectal surgery, eradicating the fistula and maintaining continence are still complex challenges for a colorectal surgeon. A minimally invasive method using a novel device was performed to consecutively treat 14 patients with anal fistula from August 2008 to November 2009. After a follow-up period of 36 months, 13 patients achieved successful closure of their fistula tracts, and recurrence occurred only in one patient. Recurrence was due to the delay of dressing change. No patient had interference with continence, and no major intra- and postoperative complications were identified. Using the novel device with invasive methods can be a promising alternative for managing anal fistulas.

Introduction Hutchinson-Gilford progeria syndrome is a rare pediatric genetic syndrome with an incidence of one per eight million live births. The disorder is characterized by premature aging, generally leading to death due to myocardial infarction or stroke at approximately 13.4 years of age. The genetic diagnosis and special clinical manifestation in two Han Chinese siblings observed at our clinic for genetic counseling are described in this report. We screened the LMNA gene in these two siblings as well as in their unaffected parents. A homozygous mutation R527C was identified in the affected siblings, and both parents were heterozygous for this variant. Case presentation In case 1, the elder 10-year-old female sibling showed the classic physical and radiological changes of Hutchinson-Gilford progeria syndrome in addition to a considerable overlap with the phenotype of mandibuloacral dysplasia. In case 2, the younger male sibling had begun to show some early physical changes at age six months. Conclusion The phenotypic findings in the patients we describe here widen the clinical spectrum of Hutchinson-Gilford progeria syndrome symptoms, providing further recognition of the phenotypic range of LMNA-associated diseases.

The capD gene, encoding a polysaccharide biosynthesis protein, was identified previously as a differential gene between Haemophilus parasuis virulent strain Nagasaki and avirulent strain SW114; however, the characteristics of this gene associating with the pathogenicity of H. parasuis remain unclear. Here, the capD deletion mutant (ΔcapD) and its complement strain (C-capD) were generated in H. parasuis virulent strain SH0165. The deletion of capD gene significantly attenuated the pathogenicity of the SH0165 strain, while the complementation of this gene largely recovered the pathogenicity to piglets. Additionally, the ΔcapD strain could not be recovered from piglets after challenge, while both SH0165 and C-capD strains were recovered from most systemic sites. Moreover, the ΔcapD strain exhibited an extreme sensitivity to the complement-mediated killing compared with SH0165 strain, while its serum-resistance ability largely restored with the capD gene complementation. These data present the evidence that the capD gene is a novel pathogenicity-associated determinant and involved in serum-resistance ability of H. parasuis.

Lentinan (β-(1→3)-D-glucan) was found to be successfully fractionated by the mixture of dimethyl sulfoxide (DMSO) and lithium chloride (LiCl) as a solvent and acetone as a precipitant. Light scattering and viscosity measurements were made on solutions of fractionated samples in pure DMSO and 0.2M LiCl/DMSO in the range of the molecular weight M(w) from 21.7 × 10(4) to 84.7 × 10(4) . The values of M(w) in both solvents were almost the same, but the remarkable difference between the values of intrinsic viscosity [η] demonstrated that the LiCl/DMSO solvent greatly enhances the stiffness of the lentinan backbone. The observed intrinsic viscosity [η] was analyzed by the Yoshizaki-Nitta-Yamakawa theory of a worm-like chain, and the persistence length q and molecular weight per unit contour length M(L) were determined roughly as 6.0 nm and 890 g nm(-1) in 0.2M LiCl/DMSO, and 5.1 nm and 890 g nm(-1) in pure DMSO, respectively. This slightly larger persistent length in 0.2M LiCl/DMSO also confirmed the higher stiffness of lentinan enhanced by the LiCl/DMSO solvent. The enhancement of the chain stiffness was ascribed to the electrostatic repulsion because of the hydrogen bonding of the hydroxyl protons of lentinan with the chloride ion, which is in turn associated with the Li(+) (DMSO)(n) macrocation complex. © 2012 Wiley Periodicals, Inc. Biopolymers 97: 840-845, 2012.

Although flexion cervical MRI has been recommended for the diagnosis of Hirayama disease (HD), no study focused on the MR features at different neck flexion angles. Moreover, no uniform flexion angle has been confirmed in clinical practice. The purpose of this study is to quantitatively investigate the MRI typical signs of HD patients in different neck flexion degree and gives a suggestion to the MR scanning. Cervical MRI in neutral and different flexion positions (cervical flexion angle 20°, 25°, 30°, 35°, and 40°) were performed in 45 HD patients. Three MRI features including anterior shifting of the posterior wall of the cervical dural canal (ASD), widening of cervical epidural space, and epidural flow voids (EFV) at each flexed position were summarized. To evaluate ASD quantitatively, the widest cervical epidural space with the maximum sagittal diameters (d) and cervical canal sagittal diameter (D) at the same level were measured. The d/D values at different angles were calculated and compared. ASD was demonstrated in 34 out of 45 cases (75.6%) at 20° and in all cases (100%) at other 4 angles (χ (2) = 25.728, P < 0.05). Significant difference was demonstrated for the appearance rate of EFV (mean 72.8%) among different angles (χ (2) = 11.373, P = 0.021). The peak mean d/D value was found at 35°. Neck flexion angles have effects on ASD, widening of cervical epidural space and EFV. 25° is recommended as the least effective diagnostic flexion angle for MRI diagnosis of HD, and 35° may be the best one.

The objective of the present study was to establish a valid transformation method of Haemophilus parasuis, the causative agent of Glasser's disease in pigs, using a novel H. parasuis-Escherichia coli shuttle vector. A 4.2 kb endogenous plasmid pYC93 was extracted from an H. parasuis field isolate and completely sequenced. Analysis of pYC93 revealed a region approximately 800 bp showing high homology with the defined replication origin oriV of pLS88, a native plasmid identified in Haemophilus ducreyi. Based on the origin region of pYC93, E. coli cloning vector pBluescript SK(+) and the Tn903 derived kanamycin cassette, a shuttle vector pSHK4 was constructed by overlapping PCR strategy. When electroporation of the 15 H. parasuis serovar reference strains and one clinical isolate SH0165 with pSHK4 was performed, only one of these strains yielded transformants with an efficiency of 8.5 x 10(2) CFUhlg of DNA. Transformation efficiency was notably increased (1.3 x 10(5) CFU/mu g of DNA) with vector DNA reisolated from the homologous transformants. This demonstrated that restriction-modification systems were involved in the barrier to transformation of H. parasuis. By utilizing an in vitro DNA modification method with cell-free extracts of the host H. parasuis strains, 15 out of 16 strains were transformable. The novel shuttle vector pSHK4 and the established electrotransformation method constitute useful tools for the genetic manipulation of H. parasuis to gain a better understanding of the pathogen. (C) 2011 Elsevier B.V. All rights reserved.

Background: Dantrolene, a specific agent for the treatment of malignant hyperthermia, was found to inhibit Ca2+ leak through not only the skeletal ryanodine receptor (RyR1), but also the cardiac ryanodine receptor (RyR2) by correcting the defective inter-domain interaction between N-terminal (1-619 amino acid) and central (2,000-2,500 amino acid) domains of RyRs. Here, the in vivo anti-arrhythmic effect of dantrolene in a human catecholaminergic polymorphic ventricular tachycardia (CPVT)-associated RyR2R2474S/+ knock-in (KI) mouse model was investigated. Methods and Results: ECG was monitored in KI mice (n=6) and wild-type (WT) mice (n=6), before and after an injection of epinephrine (1.0mg/kg) or on exercise using a treadmill. In all KI (but not WT) mice, bi-directional ventricular tachycardia (VT) was induced after an injection of epinephrine or on exercise. Pre-treatment with dantrolene (for 7-10 days) significantly inhibited the inducible VT (P

Purpose Cervical flexion movement is supposed to play an important role in the pathogenesis of Hirayama disease. But there is no report on the range of cervical flexed motion in this disease. The purpose of current study was to compare the range of cervical flexed motion in patients with Hirayama disease with the one in healthy controls using conventional lateral flexion radiographs of the cervical spine, and to investigate the diagnostic value of radiographs for Hirayama disease. Materials and methods This prospective study was approved by a local institutional review board and written informed consent was obtained. From August 2007 to March 2009, conventional flexion cervical radiographs and flexion cervical MRI were performed on 31 Hirayama disease patients (all men, age range, 16–24 years, mean age, 19.52 ± 2.29 years) and 40 control subjects (all men, age range, 16–25 years, mean age, 23.10 ± 2.20 years). The segmental and overall range of cervical flexed motion was quantitatively measured. Receiver operating characteristic curve was calculated and the diagnostic accuracy was qualified by using the area under the curve Az. Results Both the segmental and overall range of cervical flexed motion of patients determined by flexion cervical radiographs was greater than that of controls. The Az value was 0.90, indicating a moderate to good ability of conventional flexion radiographs in the diagnosis of Hirayama disease. Conclusion Hirayama disease patients have an increased flexed motion range of cervical spine which would contribute to the pathophysiological change and determine its treatment. Conventional flexion radiographs might be suitable to be used as first line radiographic examination, followed by MRI in cases of suspected Hirayama disease.