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The abnormal accumulation of fused in sarcoma (FUS) is a pathological hallmark in a proportion of patients with frontotemporal dementia and amyotrophic lateral sclerosis.

Amyloid protein cross-seeding is a peculiar phenomenon of cross-spreading among different diseases. Unlike traditional infectious ones, diseases caused by amyloid protein cross-seeding are spread by misfolded proteins instead of pathogens. As a consequence of the interactions among misfolded heterologous proteins or polypeptides, amyloid protein cross-seeding is considered to be the crucial cause of overlapping pathological transmission between various protein misfolding disorders (PMDs) in multiple tissues and cells. Here, we briefly review the phenomenon of cross-seeding among amyloid proteins. As an interesting example worth mentioning, the potential links between the novel coronavirus pneumonia (COVID-19) and some neurodegenerative diseases might be related to the amyloid protein cross-seeding, thus may cause an undesirable trend in the incidence of PMDs around the world. We then summarize the theoretical models as well as the experimental techniques for studying amyloid protein cross-seeding. Finally, we conclude with an outlook on the challenges and opportunities for basic research in this field. Cross-seeding of amyloid opens up a new perspective in our understanding of the process of amyloidogenesis, which is crucial for the development of new treatments for diseases. It is therefore valuable but still challenging to explore the cross-seeding system of amyloid protein as well as to reveal the structural basis and the intricate processes.

RNA interference is a promising way to treat cancer and the construction of a stable drug delivery system is critically important for its application. Gelatin nanospheres (GNs) comprise a biodegradable drug vehicle with excellent biocompatibility, but there are limited studies on its delivery and role in the stabilization of miRNA and siRNA. Breast cancer is the most diagnosed type of female cancer worldwide. Abnormal miRNA expression is closely related to the occurrence and progression of estrogen receptor-positive (ER+) breast cancer. In this study, miR-4458 was upregulated in ER+ breast cancer and could inhibit MCF-7 cell viability, colony formation, migration, and invasion. Collagen type XI alpha 1 (COL11A1) was identified as a directly interacting protein of miR-4458 and an important component of the extracellular matrix. High COL11A1 expression was positively correlated with poor prognosis, lower overall survival, disease-free survival, and a late tumor-node-metastasis stage. COL11A1 knockdown could inhibit MCF-7 cell migration and invasion. GNs were used to load a miR-4458 mimic or COL11A1 siRNA (si-COL11A1) to achieve sustained and controlled release in xenograft nude mice. Their tumor volume was decreased, tumor cell apoptosis was promoted, and hepatic metastasis was significantly inhibited. Moreover, the DDR2/SRC signaling pathway was inactivated after transfection with the miR-4458 mimic and si-COL11A1. In conclusion, GNs can be potentially used to deliver siRNA or miRNA, and miR-4458 and COL11A1 can be possible targets for ER+ breast cancer treatment.

The self-assembly of biomacromolecules is an extremely important process. It is potentially useful in the fields of life science and materials science. To carry out the study on the self-assembly of proteins, it is necessary to find out the suitable self-assembly conditions, which have always been a challenging task in practice. Inspired by the screening technique in the field of protein crystallization, we proposed using the same screening technique for seeking suitable protein self-assembly conditions. Based on this consideration, we selected 5 proteins (β-lactoglobulin, hemoglobin, pepsin, lysozyme, α-chymotrypsinogen (II) A) together with 5 screening kits (IndexTM, BML, Morpheus, JCSG, PEG/Ion ScreenTM) to investigate the performance of these crystallization screening techniques in order to discover new optimized conditions of protein self-assembly. The screens were all kept at 293 K for certain days, and were analyzed using optical microscope, scanning electron microscope, transmission electron microscope, atomic force microscope, fluorescence microscope, and atomic absorption spectroscope. The results demonstrated that the method of protein crystallization screening can be successfully applied in the screening of self-assembly conditions. This method is fast, high throughput, and easily implemented in an automated system, with a low protein consumption feature. These results suggested that such strategy can be applied to finding new conditions or forms in routine research of protein self-assembly. KEY POINTS: • Protein crystallization screening method is successfully applied in the screening of self-assembly conditions. • This screening method can be applied on various kinds of proteins and possess a feature of low protein consumption. • This screening method is fast, high throughput, and easily implemented in an automated system.

Liquid-liquid phase separation (LLPS) is a ubiquitous process found in a variety of fields. It is of particular importance in biological sciences since it plays essential and vital roles in a number of physiological and pathological processes in biological systems. After LLPS, the dense droplets can grow in size via incorporating solutes from surrounding environment and in some cases coalescing with other droplets. Interestingly, the size of the dense droplets seems to have an upper limit but the mechanism remains to be explored. Since the droplet size can be essential for biological functioning, it is important to understand the size evolution of the phase separated droplets. Here we propose a physical mechanism with consideration of impurities on the surface of the dense liquid droplets. The theoretical predictions can be used to explain the observations on the size evolution. Based on the coalescence mechanism, we succeeded for the first time performing a challenging task, i.e., growing a single suspended protein crystal via merging phase-separated droplets in a fully non-contact manner. The mechanism observed may be considered as a basic model for researches in much broader fields involving phase separation, such as in biology, materials science, physics, chemistry, and meteorology.

BackgroundTaiwan Biobank (TWB) project has built a nationwide database to facilitate the basic and clinical collaboration within the island and internationally, which is one of the valuable public datasets of the East Asian population. This study provided comprehensive genomic medicine findings from 1,496 WGS data from TWB.MethodsWe reanalyzed 1,496 Illumina-based whole genome sequences (WGS) of Taiwanese participants with at least 30X depth of coverage by Sentieon DNAscope, a precisionFDA challenge winner method. All single nucleotide variants (SNV) and small insertions/deletions (Indel) have been jointly called and recalibrated as one cohort dataset. Multiple practicing clinicians have reviewed clinically significant variants.ResultsWe found that each Taiwanese has 6,870.7 globally novel variants and classified all genomic positions according to the recalibrated sequence qualities. The variant quality score helps distinguish actual genetic variants among the technical false-positive variants, making the accurate variant minor allele frequency (MAF). All variant annotation information can be browsed at TaiwanGenomes (https://genomes.tw). We detected 54 PharmGKB-reported Cytochrome P450 (CYP) genes haplotype-drug pairs with MAF over 10% in the TWB cohort and 39.8% (439/1103) Taiwanese harbored at least one PharmGKB-reported human leukocyte antigen (HLA) risk allele. We also identified 23 variants located at ACMG secondary finding V3 gene list from 25 participants, indicating 1.67% of the population is harboring at least one medical actionable variant. For carrier status of all known pathogenic variants, we estimated one in 22 couples (4.52%) would be under the risk of having offspring with at least one pathogenic variant, which is in line with Japanese (JPN) and Singaporean (SGN) populations. We also detected 6.88% and 2.02% of carrier rates for alpha thalassemia and spinal muscular atrophy (SMA) for copy number pathogenic variants, respectively.ConclusionAs WGS has become affordable for everyone, a person only needs to test once for a lifetime; comprehensive WGS data reanalysis of the genomic profile will have a significant clinical impact. Our study highlights the overall picture of a complete genomic profile with medical information for a population and individuals.

Graphitic carbon nitride (g-C3N4) is a visible light-responsive photocatalytic material with important application prospects in many fields. However, the interaction between g-C3N4 monolayers makes it easy to aggregate and precipitate in aqueous solutions, and it is necessary to prepare stable g-C3N4 aqueous dispersions for their applications. Here we propose a facile, green, and low-cost method for the preparation of stable g-C3N4 dispersions by ultrasonicating g-C3N4 in lysozyme (LYZ) solution. The LYZ was adsorbed on the surface of g-C3N4 through non-covalent interactions such as electrostatic interaction, hydrogen bonding and π-cation interaction to prevent the aggregation of g-C3N4 nanolayers. The LYZ/g-C3N4 could quickly re-form a uniform aqueous dispersion solution after freeze-drying, and exhibit good stability. Further, the results of photocatalytic sterilization showed that the assisted dispersion of LYZ enhanced the bactericidal activity of g-C3N4 and exhibited promising application prospects in the field of biomedicine and water disinfection.

Triple-negative breast cancer (TNBC) is the most malignant subtype of breast cancer. Some microRNAs (miRNAs) were abnormally expressed in TNBC, and they are closely related to the occurrence and progression of TNBC. Here, we found that miR-506 was significantly downregulated in TNBC and relatively lower miR-506 expression predicted a poorer prognosis. Moreover, we found that miR-506 could inhibit MDA-MB-231 cell viability, colony formation, migration, and invasion, and suppress the ERK/Fos oncogenic signaling pathway through upregulating its direct target protein proenkephalin (PENK). Therefore, miR-506 was proposed as a nucleic acid drug for TNBC therapy. However, miRNA is unstable in vivo, which limiting its application as a therapeutic drug via conventional oral or injected therapies. Here, a gelatin nanosphere (GN) delivery system was applied for the first time to load exogenous miRNA. Exogenous miR-506 mimic was loaded on GNs and injected into the in situ TNBC animal model, and the miR-506 could achieve sustained and controlled release. The results confirmed that overexpression of miR-506 and PENK in vivo through loading on GNs inhibited in situ triple-negative breast tumor growth and metastasis significantly in the xenograft model. Moreover, we indicated that the ERK/Fos signaling pathway was intensively inactivated after overexpression of miR-506 and PENK both in vitro and in vivo, which was further validated by the ERK1/2-specific inhibitor SCH772984. In conclusion, this study demonstrates that miR-506-loaded GNs have great potential in anti-TNBC aggressiveness therapy.

To compare the effects and safety of using microwave ablation (MWA) and surgical resection for the treatment of benign thyroid nodules (BTNs) under ultrasonic guidance and investigate the effects of this treatment on stress response. Patients with BTNs were divided into the MWA and operation groups (72 cases each). Interleukin (IL)-6, IL-8, serum tumor necrosis factor (TNF-α), and hydrostatic visual analog scale (VAS) prior to the operation, at 6 h, 24 h, and 72 h post-operation were compared between the two groups. Operation times, hospitalization times, hospitalization expenses, and postoperative complications in the two groups were also compared. All patients underwent routine ultrasound and thyroid function testing at 3 and 6 months post-operation for assessment of nodule changes and thyroid hormone levels. Compared to the MWA group, the operation group had longer average operation times, longer hospital stays, a higher rate of neck pain after surgery, and a higher rate of fever (P

Relative stability of mineral elements in tissues is necessary for health. High static magnetic fields (HiSMFs) have been widely used in biomedical research and industry. However, the bioeffect of HiSMFs on animals is still unclear. In this study, we investigated the effects of HiSMF exposure on the levels of Mg, Fe, Zn, Ca, and Cu in the main organs of mice. The 8-week male C57BL/6 mice were treated by 2–4 T, 6–8 T, 10–12 T HiSMFs for 28 days. The mass fractions of Mg, Fe, Zn, Ca, and Cu in the liver, brain, kidney, and heart in mice were respectively measured by atomic absorption spectroscopy, and used to evaluate mineral element content in tissues. The 2–4 T HiSMF exposure has increased the Mg, Fe, and Ca content in the kidney, as well as the Zn content in the brain. The 6–8 T HiSMF exposure has increased the Zn level in the liver; Mg, Fe, and Ca levels in the kidney; and Fe level in the heart, while the Zn in the kidney, and Zn and Ca in the heart was decreased by 6–8 T HiSMF exposure. For the 10–12 T HiSMF exposure, the Mg in the kidney, the Fe in the liver and kidney, and Cu in the brain have been increased significantly. However, the Zn in the kidney and the Ca in the brain and the heart were reduced by 10–12 T HiSMF exposure. The HiSMF exposure for 28 days can alter the Mg, Fe, Zn, Ca, and Cu content in mice, and change with the different magnetic flux density of HiSMFs (2–4 T, 6–8 T, 10–12 T), elements, and organ types.

Large gradient high magnetic field (LG-HMF) is a powerful tool to study the effects of altered gravity on organisms. In our study, a platform for the long-term culture of aquatic organisms was designed based on a special superconducting magnet with an LG-HMF, which can provide three apparent gravity levels (µ g, 1 g, and 2 g), along with a control condition on the ground. Planarians, Dugesia japonica, were head-amputated and cultured for 5 days in a platform for head reconstruction. After planarian head regeneration, all samples were taken out from the superconducting magnet for a behavioral test under geomagnetic field and normal gravity conditions. To analyze differences among the four groups, four aspects of the planarians were considered, including head regeneration rate, phototaxis response, locomotor velocity, and righting behavior. Data showed that there was no significant difference in the planarian head regeneration rate under simulated altered gravity. According to statistical analysis of the behavioral test, all of the groups had normal functioning of the phototaxis response, while the planarians that underwent head reconstruction under the microgravity environment had significantly slower locomotor velocity and spent more time in righting behavior. Furthermore, histological staining and immunohistochemistry results helped us reveal that the locomotor system of planarians was affected by the simulated microgravity environment. We further demonstrated that the circular muscle of the planarians was weakened (hematoxylin and eosin staining), and the epithelial cilia of the planarians were reduced (anti-acetylated tubulin staining) under the simulated microgravity environment. Bioelectromagnetics. 2018;39:428-440. © 2018 Wiley Periodicals, Inc.

Objective: Magnetic drug targeting (MDT) has attracted a lot of attention in recent years as a treatment to reduce side effects and improve efficacy. To realize successful MDT, a magnet system that can create suitable magnetic field is necessary. However, the existing technology has some shortcomings such as low targeting efficiency and unsatisfactory targeting effect. The method of using time-varying magnetic field proposed in this paper provides a new solution to this problem. Methods: In this work, a permanent magnet system providing rotational magnetic field (a time-varying magnetic field) was designed and constructed. Focusing experiments of Fe3O4 particles were carried out in this system, and the mechanism of the focusing was discussed via a simplified model. Results: By using the rotational magnetic field, superparamagnetic Fe3O4 particles can be successfully driven to the designated site within a short time. Our work showed that the time-varying magnetic field can drive the superparamagnetic particles to a focusing site, which is not possible under the same magnetic field without rotation, and the aggregation speed and effect are better than those under static field. Conclusion and Significance: Our results indicate that time-varying magnetic field can be a good choice for designing magnet systems for MDT. Moreover, the idea of using time-varying magnetic field for focusing superparamagnetic particles may be applicable in non-contact processings or manipulations of paramagnetic, superparamagnetic, or even ferromagnetic materials or objects in preparation of novel materials, robotic control of objects, and so on.

Polyuria and polydipsia are the characteristics of congenital nephrogenic diabetes insipidus (CNDI). Approximately 90% of all patients with CNDI have X-linked hereditary disease, which is due to a mutation of the arginine vasopressin receptor 2 ( AVPR2) gene. This case report describes a 54-year-old male with polyuria and polydipsia and several male members of his pedigree who had the same symptoms. The proband was diagnosed with diabetes insipidus using a water-deprivation and arginine vasopressin stimulation test. Genomic DNA from the patient and his family members was extracted and the AVPR2 gene was sequenced. A novel missense mutation of a cytosine to guanine transition at position 972 (c.972C > G) was found, which resulted in the substitution of isoleucine for methionine at amino acid position 324 (p.I324M) in the seventh transmembrane domain of the protein. The proband’s mother and daughter were heterozygous for this mutation. The novel mutation of the AVPR2 gene further broadens the phenotypic spectrum of the AVPR2 gene.

We analyzed the rules of Metasequoia glyptostroboides along with latitude, including leaf length, leaf width, leaf perimeter, leaf area, ratio of leaf length to width, specific leaf area (SLA), and leaf dry mass based on eight stands growing at different latitudes in the coastal area of eastern China, as well as their relationships with climatic and soil factors. The results showed that the leaf length, leaf width and leaf perimeter increased with increasing latitude, while the leaf area and SLA firstly increased and then decreased. The mean annual temperature and annual precipitation were the major environmental factors affecting the leaf traits along latitude gradient. With the increase of soil N content, the SLA decreased firstly and then increased, while the leaf mass decreased significantly. With the increase of soil P content, the SLA increased, and the leaf mass decreased significantly.在中国东部沿海地区沿纬度梯度选取8个水杉林,研究水杉的叶片长度、叶片宽度、叶片周长、叶片面积、叶片长宽比、比叶面积、叶片干质量等叶片性状指标的纬度变化规律,分析叶片性状与气象、土壤因子的关系.结果表明: 随着纬度的升高,水杉叶片长、宽、周长逐渐增大,叶面积和比叶面积先增加后降低.在纬度梯度上,水杉叶片性状的变化主要受年均温和年均降水量的影响.随着土壤氮含量的增加,比叶面积呈先下降后上升的趋势,叶干质量则呈显著下降的趋势;随着土壤磷含量的增加,比叶面积逐渐增加,而叶干质量则显著下降.

In order to explore the variations in leaf stoichiometry based on the method of forest tree breeding, we determined the leaf carbon (C), nitrogen (N), phosphorus (P) stoichiometry among 29 Quercus acutissima provenances grown at 3 sites. The results indicated that the site (environment) effect was statistically significant on leaf stoichiometry, with 13.2%-66.7% of the total variations accounted for leaf C, N, P, C/N, C/P and N/P, while the provenance effect was insignificant and only accounted for 2.9%-11.0% of total variations for leaf stoichiometry. The leaf N and C/N, N and N/P, P and C/P, P and N/P were significantly correlated, and the common standardized major axis slope was also observed among three sites and two provenance groups. It could be concluded that the leaf stoichiometry of Q. acutissima, was mainly determined by its growing environment, due to the similar C, N and P biochemical pathways at species level. The stable correlation coefficients among sites and provenances implied the coupling ratios of leaf stoichiometry were independent of environment and provenance, which supported the leaf stoichiometric homeostasis.结合林木育种方法,以3个麻栎种源试验林为研究对象,测定了29个麻栎种源叶片碳、氮、磷化学计量特征.结果表明: 地点(环境)显著影响叶片碳、氮、磷、碳氮比、碳磷比、氮磷比,解释量占总变异量的13.2%～66.7%,而种源(遗传)的影响并不显著,解释量只占总变异量的2.9%～11.0%.叶片氮与碳氮比、氮与氮磷比、磷与碳磷比、磷与氮磷比均具有显著的相关性,且无论是地点间还是东西部种源间均存在共同的标准化主轴斜率.表明在单一树种(麻栎)水平上,种源间具有相似的碳、氮、磷生化过程,其叶片化学计量特征主要受环境的影响;而化学计量特征在地点间和东西部种源间稳定的相关系数反映出叶片化学计量特征的特定耦合比例不受环境和种源的影响,支持化学计量学的内稳性理论.

OBJECTIVE: To investigate the effect of sericin on the proliferation of human gastric cancer MKN45 cells and explore the underlying molecular mechanism. METHODS: MKN45 cells were transfected by LC3 double fluorescent autophagic virus, and the positive cells screened by puromycin were divided into blank group, sericin group and sericin+3-MA group. After incubation with sericin for 48 h, the cells were examined for proliferation, apoptosis and cell cycle using CCK-8 assay and flow cytometry. Cell autophagy was detected by transmission electron microscopy (TEM) and fluorescent inverted microscope, and the autophagy-related markers including LC3, p62 and Beclin proteins were detected with Western blotting. Nude mice bearing gastric cancer xenograft were treated with normal saline or sericin injections (n=5) and the changes in the tumor volume and weight were measured. RESULTS: Compared with the blank group, MKN45 cells with sericin treatment showed significantly inhibited proliferation both in vitro and in nude mice. Autophagosomes were observed in sericin-treated cells under TEM and fluorescent inverted microscope. Sericin treatment of the cells significantly increased the cell apoptosis (P < 0.01), caused obvious cell cycle arrest in G(2)/M phase (P < 0.01), up-regulated the expressions of both LC3-2 and Beclin, and down-regulated the expression of p62. The autophagy inhibitor 3-MA obviously antagonized the effects of sericin on cell apoptosis, cell cycle and autophagic protein expressions. CONCLUSION: Sericin can inhibit the proliferation of human gastric cancer MKN45 cells by regulating cell autophagy to serve as potential anti-tumor agent.

One well-known prerequisite for successful crystallization from solution is a supersaturated solution. To achieve supersaturation, many methods are known, among which solvent evaporation is a common approach. For protein crystallization, the most widely used method is vapor diffusion, in which solvent evaporation from the crystallization solution is the major reason for achieving supersaturation. The solvent evaporation rate may affect the actual concentration distribution in the crystallization solution, thereby influencing the crystallization process. To explore the effect of evaporation rate on protein crystallization, we used lysozyme as a model protein and studied the crystallization success rate at different evaporation conditions. Successful crystallization occurred only when both supersaturation and evaporation rates were in suitable ranges. This study demonstrates that both supersaturation level and the rate of reaching supersaturation (or solvent evaporation rate) are important for lysozyme crystallization. To increase the chance of obtaining crystals, manipulation of solvent evaporation rate is one choice. According to this assumption, we performed crystallization screening trials at different evaporation rates using three model proteins. The trials demonstrate that control of the evaporation rate during crystallization may provide more opportunities to obtain crystals.

Large gradient high magnetic field (LG-HMF) is a powerful tool to study the effects of altered gravity on organisms. In our study, a platform for the long-term culture of aquatic organisms was designed based on a special superconducting magnet with an LG-HMF, which can provide three apparent gravity levels (µ g, 1 g, and 2 g), along with a control condition on the ground. Planarians, Dugesia japonica, were head-amputated and cultured for 5 days in a platform for head reconstruction. After planarian head regeneration, all samples were taken out from the superconducting magnet for a behavioral test under geomagnetic field and normal gravity conditions. To analyze differences among the four groups, four aspects of the planarians were considered, including head regeneration rate, phototaxis response, locomotor velocity, and righting behavior. Data showed that there was no significant difference in the planarian head regeneration rate under simulated altered gravity. According to statistical analysis of the behavioral test, all of the groups had normal functioning of the phototaxis response, while the planarians that underwent head reconstruction under the microgravity environment had significantly slower locomotor velocity and spent more time in righting behavior. Furthermore, histological staining and immunohistochemistry results helped us reveal that the locomotor system of planarians was affected by the simulated microgravity environment. We further demonstrated that the circular muscle of the planarians was weakened (hematoxylin and eosin staining), and the epithelial cilia of the planarians were reduced (anti-acetylated tubulin staining) under the simulated microgravity environment. Bioelectromagnetics. 2018;39:428-440. © 2018 Wiley Periodicals, Inc.

This paper shows the corrosion of steel bar from a new perspective with viscoelasticity theory. The strain of steel bar is decomposed as initial elastic strain and creep strain, and the creep strain is caused by corrosion. In order to analysis creep strain, corrosion factor and corrosion strain function were introduced to reflect the deformation behavior. Then integral constitutive relation of corroded bar was deduced according to the above assumption. As a special case of this approach, the corrosion factor of steel bar in atmospheric environment is introduced. The results indicate that the corrosion factor increase with diameter deducing, and the deformation of the steel bar possess the acceleration creep behavior during the corrosion process.

This report describes the modification of crystallization plates by simply oxidizing the surface of the protein wells. The oxidized crystallization plates were tested in standard protein crystallization screening and reproducibility studies. The results showed that the protein wells of the treated plates were smoother and more optically transparent than those of the untreated plates, and more importantly, protein crystallization was significantly promoted after the oxidation treatment. Because there is no change to the routine screening protocol, this method is simple and easy to apply in protein crystallization.

The effect of magnetic fields on water is still a highly controversial topic despite the vast amount of research devoted to this topic in past decades. Enhanced water evaporation in a magnetic field, however, is less disputed. The underlying mechanism for this phenomenon has been investigated in previous studies. In this paper, we present an investigation of the evaporation of water in a large gradient magnetic field. The evaporation of pure water at simulated gravity positions (0 gravity level (ab. g), 1 g, 1.56 g and 1.96 g) in a superconducting magnet was compared with that in the absence of the magnetic field. The results showed that the evaporation of water was indeed faster in the magnetic field than in the absence of the magnetic field. Furthermore, the amount of water evaporation differed depending on the position of the sample within the magnetic field. In particular, the evaporation at 0 g was clearly faster than that at other positions. The results are discussed from the point of view of the evaporation surface area of the water/air interface and the convection induced by the magnetization force due to the difference in the magnetic susceptibility of water vapor and the surrounding air.

A novel form-stable capric acid/organophilic montmorillonite composite for thermal energy storage is developed in this study. The morphology and thermal properties were determined by scanning electron microscope(SEM), polarized optical microscope(POM), differential scanning calorimetry(DSC) and thermogravimetric analyzer(TGA). The DSC results showed that the on-set temperature of the sample with 40% wt CA was closed to 29°C, the latent heat was 35.8 J/g at 56 kPa and 51.5 J/g at 0 kPa, while the on-set temperature of sample with 60% CA was also 29°C, the latent heat was 79.7 J/g at 56 kPa and 80.8 J/g at 0 kPa. TG investigations revealed that the composites had excellent thermal stability above their working temperature ranges. The POM images exhibited phase behaviors of composites to confirm leakage, and the samples with 20% and 40% CA showed good thermal properties. In addition, SEM images presented the microstructure of all the samples. All of the conclusions indicated that sample with 40% wt CA was a better candidate for novel form-stable CA/OMMT composite for low-temperature thermal energy storage applications.

In order to explore the durability of high bending strength pavement concrete, adding silica flour, fibrous reticulum, SD emulsion and HG emulsion separately in concrete mixture, the shrinkage characteristic, wear resistance and fatigue property of different high bending strength pavement concrete are tested, and contrastive analysis with the plain concrete material is researched. The test results show that the early contraction of HBSPC is bigger, but the total drying shrinkage strain is smaller than plain concrete, and the drying shrinkage stable stage of HBSPC is shorter than plain concrete. The temperature shrinkage of HBSPC and plain concrete is uniform basically, the temperature shrinkage coefficient of HBSPC is lower than the plain concrete in the negative warm area and warm area, but it is higher in 0 °C neighbor. The mixtures of the material enhance the wear resistance and fatigue property of concrete.

To construct a MYH9 gene knockout model in MGC803 cell line using transcription activator-like effector nuclease (TALEN) and observe its effect on cell cycle and apoptosis.According to FastTALE(TM) TALEN Kit, we designed TALEN pairs and constructed the plasmids targeting to MYH9 gene. After detecting their activity in MGC803 cells by plasmid transfection, DNA sequencing, RT-PCR and western blot, we selected the monoclonal cells and studied the changes in the cell cycle and apoptosis.MYH9 gene could not be knocked out but knocked down in selected MGC803 monoclonal cells, which caused cell cycle arrested at G2/M phase (P0.05) and a significant increase in the cell number with early apoptosis (P0.01).We successfully generated a MYH9 knockdown model in MGC803 cell lines by TALEN, which could be in favor of MYH9 function study in gastric cancer.

Biliary atresia (BA) is characterized by the progressive fibrosclerosing obliteration of the extrahepatic biliary system during the first few weeks of life. Despite early diagnosis and prompt surgical intervention, the disease progresses to cirrhosis in many patients. The current theory for the pathogenesis of BA proposes that during the perinatal period, a still unknown exogenous factor meets the innate immune system of a genetically predisposed individual and induces an uncontrollable and potentially self-limiting immune response, which becomes manifest in liver fibrosis and atresia of the extrahepatic bile ducts. Genetic factors that could account for the disease, let alone for its high incidence in Chinese, are to be investigated. To identify BA susceptibility loci, we carried out a genome-wide association study (GWAS) using the Affymetrix 5.0 and 500 K marker sets. We genotyped nearly 500 000 single-nucleotide polymorphisms (SNPs) in 200 Chinese BA patients and 481 ethnically matched control subjects. The 10 most BA-associated SNPs from the GWAS were genotyped in an independent set of 124 BA and 90 control subjects. The strongest overall association was found for rs17095355 on 10q24, downstream XPNPEP1, a gene involved in the metabolism of inflammatory mediators. Allelic chi-square test P-value for the meta-analysis of the GWAS and replication results was 6.94 x 10(-9). The identification of putative BA susceptibility loci not only opens new fields of investigation into the mechanisms underlying BA but may also provide new clues for the development of preventive and curative strategies.

Varying the temperature has been proven to be beneficial for improving the screening efficiency of protein crystallisation, and thus a crystallisation screening strategy based on this phenomenon can be developed. Such a temperature varying strategy can be applied in practical crystallisation screening, however, there are no guidelines for determining what temperature programme should be utilised. It is therefore necessary to investigate how the temperature programme affects the crystallisation process, so as to help people design a suitable temperature programme. For this purpose, we investigated the effect of temperature programmes on the protein crystallisation (lysozyme, proteinase K, and concanavalin A) that are characterised by different solubility behaviours with respect to temperature. Judging from the reproducibility studies of protein crystallisation with different temperature programmes, we recommend using linear temperature programmes for a moderate time period (24 to 48 h) and a large temperature range according to the properties of the proteins. (© 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

Temperature is generally considered as an important factor in protein crystallization. Such is true because crystals usually grow at a preferable temperature in a certain crystallization solution. If a nonsuitable temperature is used, the solution will not yield crystals. However, it is difficult to decide the best temperature suited for screening the crystallization condition of proteins. In this study, it was found out that, compared to constant temperature, a variation in a reasonable range can result in a more efficient crystallization screening. Using the Sparse Matrix Screen with the screening kit Index, this study tested nine commercially available proteins and proved that, compared to the conventional constant temperature strategy, a varying temperature strategy can actually increase the possibility of obtaining crystals. Consequently, the cycling temperature strategy (CTS) is then proposed to be utilized in most screening tasks when the suitable crystallization temperature is unknown.

Magnetic field has been found to be able to improve the quality of some protein crystals; containerless condition is considered to be beneficial for growing good-quality crystals. Nevertheless, there has been no report on combination of these two conditions for protein crystallization. In this paper, crystallization and dissolution of protein crystals in a levitated containerless droplet in a strong magnetic field has been successfully achieved. New phenomena were observed for both crystallization and dissolution processes.

Background. The CD134/CD134L interaction provides a strong costimulatory signal that is CD28-independent. The CD134/CD134L pathway has been studied in inflammatory, autoimmune diseases, and graft-versus-host disease, but no information exists on the involvement of CD134/CD134L interactions in solid organ transplantation. Methods. CD134/CD134L costimulation was studied in a rat model of small bowel transplantation. Immunohistochemistry and flow cytometry were used to analyze the expression and localization of CD134/ CD134L. Mixed lymphocyte culture and quantitative RT-PCR were used to measure the effect on T proliferation and T helper cell cytokine transcripts of single or combined CD134 and CD28 costimulatory blockade. Results. CD134 and CD134L molecules were strongly expressed in acutely rejected small bowel allografts. This expression was significantly suppressed by FK506. Interruption of the CD134 and CD28 costimulatory pathways resulted in an pronounced increase in expression of interleukin-10 transcripts. Conclusion. These results present the first evidence that the CD134/CD134L interaction is associated with acute allograft rejection. Combined CD134/CD134L blockade may be an effective treatment to both prevent acute allograft rejection and promote the induction of cells with regulatory capacity.

High-quality crystals are key to obtaining accurate three-dimensional structures of proteins using X-ray diffraction techniques. However, obtaining such protein crystals is often a challenge. Several containerless crystallization techniques have been reported to have the ability to improve crystal quality, but it is unknown which is the most favourable way to grow high-quality protein crystals. In this paper, a quality comparison of protein crystals which were grown under three containerless conditions provided by diamagnetic levitation, silicone oil and agarose gel was conducted. A control experiment on a vessel wall was also simultaneously carried out. Seven different proteins were crystallized under the four conditions, and the crystal quality was assessed in terms of the resolution limit, the mosaicity and theRmerge. It was found that the crystals grown under the three containerless conditions demonstrated better morphology than those of the control. X-ray diffraction data indicated that the quality of the crystals grown under the three containerless conditions was better than that of the control. Of the three containerless crystallization techniques, the diamagnetic levitation technique exhibited the best performance in enhancing crystal quality. This paper is to our knowledge the first report of improvement of crystal quality using a diamagnetic levitation technique. Crystals obtained from agarose gel demonstrated the second best improvement in crystal quality. The study indicated that the diamagnetic levitation technique is indeed a favourable method for growing high-quality protein crystals, and its utilization is thus potentially useful in practical efforts to obtain well diffracting protein crystals.

BACKGROUND Small bowel allografts can either be placed heterotopically or orthotopically. In heterotopic small bowel transplantation (H-SBT), the host small intestine containing a substantial amount of gut-associated lymphoid tissue is removed, whereas in conventional orthotopic small bowel transplantation (O-SBT) it is retained. This study compared the allograft survival of H-SBT and O-SBT and evaluated the effect of retaining the host intestine in O-SBT in an altered O-SBT (AO-SBT) model. METHODS SBT was performed in a high-responder rat stain combination (blood group D Agouti --> Lewis). Immunosuppressive treatment consisting of a short course of FK506 (2 mg/kg/day IM for 3 days before transplantation and 0.3 mg/kg/day for 14 days after transplantation) was used. RESULTS Survival (mean +/- SD) of H-, O-, and AO-SBT untreated animals was 6.25+/-0.58 days, 6.5+/-0.58 days, and 6.7+/-0.25 days, respectively. With FK506 immunosuppression, survival of H-SBT animals was 49.3+/-13.17 days, whereas 75% (12/16) and 80% (4/5), respectively, of O-SBT and AO-SBT animals achieved indefinite survival (>120 days) with functioning grafts. CONCLUSION Our data suggest that heterotopic placement of intestinal allografts results in a more severe graft rejection than orthotopic placement. The indefinite survival of O-SBT is not due to of the removal of host intestine carrying a heavy load of gut-associated lymphoid tissue.

As the most widely utilized technique to determine the 3-dimensional structure of protein molecules, X-ray crystallography can provide structure of the highest resolution among the developed techniques. The resolution obtained via X-ray crystallography is known to be influenced by many factors, such as the crystal quality, diffraction techniques, and X-ray sources, etc. In this paper, the authors found that the protein sequence could also be one of the factors. We extracted information of the resolution and the sequence of proteins from the Protein Data Bank (PDB), classified the proteins into different clusters according to the sequence similarity, and statistically analyzed the relationship between the sequence similarity and the best resolution obtained. The results showed that there was a pronounced correlation between the sequence similarity and the obtained resolution. These results indicate that protein structure itself is one variable that may affect resolution when X-ray crystallography is used.

Hirschsprung's disease (HSCR), or aganglionic megacolon, is a congenital disorder characterized by the absence of enteric ganglia in variable portions of the distal intestine. RET is a well-established susceptibility locus, although existing evidence strongly suggests additional loci contributing to sporadic HSCR. To identify these additional genetic loci, we carried out a genome-wide association study using the Affymetrix 500K marker set. We successfully genotyped 293,836 SNPs in 181 Chinese subjects with sporadic HSCR and 346 ethnically matched control subjects. The SNPs most associated with HSCR were genotyped in an independent set of 190 HSCR and 510 control subjects. Aside from SNPs in RET, the strongest overall associations in plausible candidate genes were found for 2 SNPs located in intron 1 of the neuregulin 1 gene (NRG1) on 8p12, with rs16879552 and rs7835688 yielding odds ratios of 1.68 [Cl 95%:(1.40, 2.00), P= 1.80 × 10-8] and 1.98 [Cl 95%:(1.59, 2.47), P = 1.12 × 10-9], respectively, for the heterozygous risk genotypes under an additive model. There was also a significant interaction between RET and NRG1 (P = 0.0095), increasing the odds ratio 2.3-fold to 19.53 for the RET rs2435357 risk genotype (TT) in the presence of the NRG1 rs7835688 heterozygote, indicating that NRG1 is a modifier of HSRC penetrance. Our highly significant association findings are backed-up by the important role of NRG1 as regulator of the development of the enteric ganglia precursors. The identification of NRG1 as an additional HSCR susceptibility locus not only opens unique fields of investigation into the mechanisms underlying the HSCR pathology, but also the mechanisms by which a discrete number of loci interact with each other to cause disease. © 2009 by The National Academy of Sciences of the USA., link_to_OA_fulltext

Hirschsprung's disease (HSCR) is a congenital disorder in which ganglion cells are absent in variable portions of the lower digestive tract according to which patients are classified. The RET gene is the major HSCR gene, although reduced penetrance of RET mutations and variable expression of HSCR phenotype indicates that more than one gene is required. An unidentified RET-dependent modifier on 3p21 appears to be necessary for transmission of the short HSCR (S-HSCR) phenotype. We investigated 6 Mb of the 3p21 region on a quest for the HSCR-susceptibility locus. Fifty-eight S-HSCR case-parent trios were genotyped using Sequenom technology for 214 tag single nucleotide polymorphisms (SNPs) distributed along 6 Mb of the 3p21 region. A five-marker haplotype, spanning a 118 kb gene-rich region, was found to be overtransmitted to affected offspring. The associated haplotype encompasses three genes involved in neurological phenotypes. Importantly, this association was replicated in an independent sample of 172 S-HSCR cases and 153 unrelated controls. Ranking markers by proximity to candidate genes or by expected functional consequences could be used in follow-up studies to finally pinpoint this HSCR locus.

Small bowel allografts can either be placed heterotopically or orthotopically. In heterotopic small bowel transplantation (H-SBT), the host small intestine containing a substantial amount of gut-associated lymphoid tissue is removed, whereas in conventional orthotopic small bowel transplantation (O-SBT) it is retained. This study compared the allograft survival of H-SBT and O-SBT and evaluated the effect of retaining the host intestine in O-SBT in an altered O-SBT (AO-SBT) model.SBT was performed in a high-responder rat stain combination (blood group D Agouti --Lewis). Immunosuppressive treatment consisting of a short course of FK506 (2 mg/kg/day IM for 3 days before transplantation and 0.3 mg/kg/day for 14 days after transplantation) was used.Survival (mean +/- SD) of H-, O-, and AO-SBT untreated animals was 6.25+/-0.58 days, 6.5+/-0.58 days, and 6.7+/-0.25 days, respectively. With FK506 immunosuppression, survival of H-SBT animals was 49.3+/-13.17 days, whereas 75% (12/16) and 80% (4/5), respectively, of O-SBT and AO-SBT animals achieved indefinite survival (120 days) with functioning grafts.Our data suggest that heterotopic placement of intestinal allografts results in a more severe graft rejection than orthotopic placement. The indefinite survival of O-SBT is not due to of the removal of host intestine carrying a heavy load of gut-associated lymphoid tissue.

Table 2 of the article by Caoet al.[(2013),Acta Cryst.D69, 1901–1910] is corrected.

Containerless processing of materials is considered beneficial for obtaining high quality products due to the elimination of the detrimental effects coming from the contact with container walls. Many containerless processing methods are realized by levitation techniques. This paper describes a containerless levitation setup that utilized the magnetization force generated in a gradient magnetic field. It comprises a levitation unit, a temperature control unit, and a real-time observation unit. Known volume of liquid diamagnetic samples can be levitated in the levitation chamber, the temperature of which is controlled using the temperature control unit. The evolution of the levitated sample is observed in real time using the observation unit. With this setup, containerless processing of liquid such as crystal growth from solution can be realized in a well-controlled manner. Since the levitation is achieved using a superconducting magnet, experiments requiring long duration time such as protein crystallization and simulation of space environment for living system can be easily succeeded.