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Exosomes derived from hepatocellular carcinoma (HCC) cells are nanovesicles and are involved in the occurrence and development of HCC, they also serve as important carriers and drug targets of nanodrug delivery systems. The external shape and internal structure of exosomes are important indexes of identification, and isolated intact morphology is crucial to biological function integrity. However, given their susceptibility to various influencing factors, the external shape and internal structure of exosomes derived from HCC cells remain incompletely studied. In this study, exosomes purified from HCC cells were isolated at different centrifugation speeds and examined via multiple electron microscopy (EM) techniques. The results demonstrate that exosomes possess a nearly spherical shape and bilipid membranous vesicle with a concave cavity structure containing electron-dense and coated vesicles, suggesting the possible existence of subpopulations of exosomes with specific functions. The exosomes isolated at ultracentrifugation (UC) speed (≥110,000×g) presented irregular and diverse external morphologies, indicating the effect on the integrity of the exosomes. Transforming growth factor signaling bioactive substances (TGF-β1, S100A8, and S100A9) can be found in exosomes by performing Western blotting, showing that the internal content is associated with metastasis of HCC. These findings show that EMelectron microscopy and UC speed can affect exosome characteristics, including external shape, internal structure, and content of bioactive substances. The electron-dense and coated vesicles that had been discovered in exosomes might become new additional morphological features, which could help to improve the interpretation of experimental results and widen our understanding of exosome morphology.
Stem cells are an immortal cell population capable of self-renewal; they are essential for human development and ageing and are a major focus of research in regenerative medicine. Despite considerable progress in differentiation of stem cells in vitro, culture conditions require further optimization to maximize the potential for multicellular differentiation during expansion. The aim of this study was to develop a feeder-free, serum-free culture method for human embryonic stem cells (hESCs), to establish optimal conditions for hESC proliferation, and to determine the biological characteristics of the resulting hESCs. The H9 hESC line was cultured using a homemade serum-free, feeder-free culture system, and growth was observed. The expression of pluripotency proteins (OCT4, NANOG, SOX2, LIN28, SSEA-3, SSEA-4, TRA-1-60, and TRA-1-81) in hESCs was determined by immunofluorescence and western blotting. The mRNA expression levels of genes encoding nestin, brachyury and α-fetoprotein in differentiated H9 cells were determined by RT-PCR. The newly developed culture system resulted in classical hESC colonies that were round or elliptical in shape, with clear and neat boundaries. The expression of pluripotency proteins was increased, and the genes encoding nestin, brachyury, and α-fetoprotein were expressed in H9 cells, suggesting that the cells maintained in vitro differentiation capacity. Our culture system containing a unique set of components, with animal-derived substances, maintained the self-renewal potential and pluripotency of H9 cells for eight passages. Further optimization of this system may expand the clinical application of hESCs.
To explore whether different polyvinylpyrrolidone (PVP) concentrations affect the results of intracytoplasmic sperm injection (ICSI), a prospective study was conducted for 194 couples undergoing 210 ICSI therapy cycles. These cycles were divided into three groups (10, 7 and 5% groups) using the corresponding concentration of PVP for sperm immobilization. The main outcome measures were analyzed. Results indicated that, with a decrease in PVP concentrations, all of the main outcome measures increased. In particular, the high-quality cleavage embryo rate in the 7% group was significantly lower than in the 5% group (P < 0.01), and the cleavage, high-quality cleavage embryo, and high-quality blastocyst rates in the 5% group were significantly higher than those in the 10% group (all P < 0.001). For high-/intermediate-quality semen, all of the main outcome measures were significantly increased with 5% PVP. For the poor-quality semen, only the high-quality cleavage embryo and high-quality blastocyst rates were significantly higher in the 5% group. Therefore, lowering PVP concentrations greatly promoted the development of embryos in ICSI cycles, with an optimal concentration of 5% for ICSI.
This paper focus on the mechanical and martensitic transformation behaviors of axially functionally graded shape memory alloy (AFG SMA) beams. It is taken into consideration that material properties, such as austenitic elastic modulus, martensitic elastic modulus, critical transformation stresses and maximum transformation strain vary continuously along the longitudinal direction. According to the simplified linear SMA constitutive equations and Bernoulli-Euler beam theory, the formulations of stress, strain, martensitic volume fraction and governing equations of the deflection, height and length of transformed layers are derived. Employing the Galerkin’s weighted residual method, the governing differential equation of the deflection is solved. As an example, the bending behaviors of an AFG SMA cantilever beam subjected to an end concentrated load are numerically analyzed using the developed model. Results show that the mechanical and martensitic transformation behaviors of the AFG SMA beam are complex after the martensitic transformation of SMA occurs. The influences of FG parameter on the mechanical behaviors and geometrical shape of transformed regions are obvious, and should be considered in the design and analysis of AFG SMA beams in the related regions.
Animal studies have suggested that mushroom intake can alleviate non-alcoholic fatty liver disease (NAFLD) due to its anti-inflammatory and antioxidant properties. However, the association between mushroom intake and NAFLD is unknown in humans. We aimed to investigate the association of mushroom intake with NAFLD among Chinese adults. This is a cross-sectional study of 24 236 adults (mean (standard deviation) age: 40·7 (sd 11·9) years; 11 394 men (47·0 %)). Mushroom intake was assessed via a validated FFQ. Newly diagnosed NAFLD was identified based on the results of annual health examinations, including ultrasound findings and a self-reported history of the disease. Multiple logistic models were used to examine the association between mushroom intake and NAFLD. The prevalence of newly diagnosed NAFLD was 19·0 %. Compared with those consuming mushrooms less frequently (≤1 time/week), the fully adjusted OR of newly diagnosed NAFLD were 0·95 (95 % CI 0·86, 1·05) for those consuming 2–3 times/week and 0·76 (95 % CI 0·63, 0·92) for those consuming ≥4 times/week (Pfor trend = 0·01). The inverse association was consistent in subgroups defined by age, sex and BMI. In conclusion, higher mushroom intake was significantly associated with lower prevalence of NAFLD among Chinese adults. Future research is required to understand the causal association between mushroom intake and NAFLD.
The aim of this study is to describe the risk factors of injured children’s posttraumatic stress disorder (PTSD) in China and provide the scientific data for PTSD prevention and control in children.
Electronic databases, including Medline, Science Direct, Google Scholar, and CNKI (China National Knowledge Infrastructure), were searched for articles published on or before October 30, 2018, searching for the words, “PTSD,” “child”/“children,” “injury”/“injuries,” and “China”/“Chinese.” Forty-seven articles met the inclusion criteria and were meta-analyzed using random-effect models.
The total sample size in our meta-analysis was 65 298, and there were 13 402 children diagnosed with PTSD. The prevalence of PTSD after suffering from injury was 20.52% (95% CI = 17%–23%). PTSD occurrence was higher in girls than boys (24.61% vs 19.36%, P < 0.001). The PTSD rate for students was 51.82%, 37.12%, and 14.02% in senior, junior high school children, and primary school student, respectively. PTSD prevalence was 58.93% in rural children and higher than the urban children (57.36%). The prevalence of PTSD in ethnic minority children was significantly higher than that of Han Chinese children (35.38% vs 13.50%).
PTSD in injured children is significantly higher in girls, senior high school children, in rural areas, and in ethnic minority children. PTSD prevention and control should be focused on these 4 subgroups.
p53, DNA damage regulated gene (PDRG) and apoptosis-stimulating p53 protein 2 (ASPP2) are p53-related genes which can promote apoptosis. The full-length cDNA sequence of the Px-pdrg and Px-aspp2 genes were characterized and their mRNA expression dynamics under heat stress were studied in diamondback moth (DBM) Plutella xylostella collected from Fuzhou, China. The full-length cDNA of Px-pdrg and Px-aspp2 spans 721 and 4201 bp, containing 395 and 3216 bp of the open reading frame, which encode a putative protein comprising 130 and 1072 amino acids with a calculated molecular weight of 14.58 and 118.91 kDa, respectively. As compared to 25°C, both Px-pdrg and Px-aspp2 were upregulated in chlorpyrifos-resistant (Rc) and -susceptible (Sm) strains of DBM adults and pupae under heat stress. In addition, Rc DBM showed a significantly higher expression level of Px-pdrg and Px-aspp2 in contrast to Sm DBM. The results indicate that high temperature can significantly promote apoptosis process, especially in Rc-DBM. Significant fitness cost in Rc-DBM might be associated with drastically higher transcript abundance of Px-pdrg and Px-aspp2 under the heat stress.
Iron sulfides have attracted much interests for their potential as anode materials in energy storage devices in view of their low costs, and environmentally benign and high theoretical capacities. Among them, Fe1−xS is relatively rarely investigated. In this work, Fe1−xS@rGO has been synthesized using a facile in situ hydrothermal method. After wrapped by rGO, the morphology of Fe1−xS particles changes from hexagonal flakes to irregular particles with much smaller sizes. As the anode material for lithium ion batteries, Fe1−xS@rGO exhibits excellent lithium storage ability. It can deliver an initial discharge capacity of 1575.5 mA h/g in the potential window of 0.005–3 V, and a reversible capacity of 907.8 mA h/g can be maintained after 200 cycles at 100 mA/g. Its improved electrochemical performance can be attributed to the effect of enhanced contact area and shortened Li+ ion transport distance because of rGO’s contribution.
Cinnamomum chago is a woody species of the family Lauraceae endemic to Yunnan province, China, previously known from only one location, and categorized as a Plant Species with Extremely Small Population. We surveyed to determine the distribution and population size of C. chago, characterize its habitat, identify any threats, assess its conservation status, and provide guidelines for its management and conservation. During 2014–2017 we found only 64 mature C. chago, in five locations. These small, fragmented populations occur along Lancang River in Dali Prefecture at altitudes of 2,200–2,400 m. The species' extent of occurrence is c. 923 km2, with an area of occupancy of c. 60 km2. The habitat of the species has been degraded by expansion of pastoral activities and deforestation. We recommend categorization of C. chago as Endangered on the IUCN Red List, prevention of the collection of seeds and wood of the species, protection and monitoring, and ex situ propagation for future reintroductions.
The present study aimed to evaluate the validity and reproducibility of a thirteen-item FFQ regarding identification of dietary conditions in a rural population in China.
A reproducibility study repeated the first FFQ (FFQ1) approximately 4 weeks later (FFQ2). A validity study evaluated the mean of three consecutive 24 h diet recalls as the reference measure.
Residents of a rural area in Henan Province, which is located in the central region of China.
A total of 295 individuals participated in the reproducibility study. In addition, 123 people agreed to participate in the validity study. Spearman’s correlation coefficients between the two FFQ ranged from 0·06 (vegetables) to 0·58 (eggs). Spearman’s correlation coefficients between the two methods of collection ranged from 0·01 for cereal to 0·49 for staple foods. The mean of the intraclass correlation coefficients of the two FFQ (FFQ1 v. FFQ2) was 0·19. Bland–Altman analysis indicated good agreement for most food groups across the range of intake for the two studies.
The study demonstrated that our FFQ design could be used as a representative tool to conduct a dietary evaluation of a rural population.
Several observational studies have investigated the association of insomnia with psychiatric disorders. Such studies yielded mixed results, and whether these associations are causal remains unclear. Thus, we aimed to identify the causal relationships between insomnia and five major psychiatric disorders.
The analysis was implemented with six genome-wide association studies; one for insomnia and five for psychiatric disorders (attention-deficit/hyperactivity disorder, autism spectrum disorder, major depressive disorder, schizophrenia, and bipolar disorder). A heterogeneity in dependent instrument (HEIDI) approach was used to remove the pleiotropic instruments, Mendelian randomization (MR)-Egger regression was adopted to test the validity of the screened instruments, and bidirectional generalized summary data-based MR was performed to estimate the causal relationships between insomnia and these major psychiatric disorders.
We observed significant causal effects of insomnia on the risk of autism spectrum disorder and bipolar disorder, with odds ratios of 1.739 (95% confidence interval: 1.217–2.486, p = 0.002) and 1.786 (95% confidence interval: 1.396–2.285, p = 4.02 × 10−6), respectively. There was no convincing evidence of reverse causality for insomnia with these two disorders (p = 0.945 and 0.546, respectively). When insomnia was considered as either the exposure or outcome variable, causal estimates for the remaining three psychiatric disorders were not significant.
Our results suggest a causal role of insomnia in autism spectrum disorder and bipolar disorder. Future disease models should include insomnia as a factor for these two disorders to develop effective interventions. More detailed mechanism studies may also be inspired by this causal inference.
Many components in diet have regulated oxidative stress, inflammatory reaction and even balance oestrogen levels. Because these factors are closely associated with depressive symptoms in postmenopausal women, it is considered that dietary factors are able to prevent and control depressive symptoms. On the other hand, a dietary pattern that considers the correlations and synergies between foods and nutrients is expected to have a greater impact on disease risk. The aim of the present study is to evaluate whether dietary patterns are associated with depressive symptoms in Chinese postmenopausal women. A cross-sectional study of 2051 postmenopausal women (mean age: 58·8 (sd 7·4) years) was conducted in Tianjin, China. Dietary consumption was assessed by a valid self-administered FFQ. Principal component analysis was used to derive three major dietary patterns: ‘healthy’, ‘sweets’ and ‘traditional Tianjin’ from eighty-eight food items. Depressive symptoms were assessed using the Zung Self-Rating Depression Scale, and cut-off point of 48 indicating serious depressive symptoms. The association between quartile of dietary patterns and depressive symptoms was assessed using multiple logistic regression analysis. The multivariable-adjusted OR of having depressive symptoms for increasing quartile of dietary patterns were as follows: healthy, 1·00, 0·79 (95 % CI 0·49, 1·28), 0·62 (95 % CI 0·37, 1·04) and 0·57 (95 % CI 0·33, 0·97); sweets, 1·00, 0·75 (95 % CI 0·42, 1·3), 1·08 (95 % CI 0·64, 1·81) and 1·66 (95 % CI 1·03, 2·71); and traditional Tianjin, 1·00, 1·02 (95 % CI 0·58, 1·79), 0·96 (95 % CI 0·54, 1·71) and 2·53 (95 % CI 1·58, 4·16), respectively. The present study demonstrated that a healthy dietary pattern was inversely associated with depressive symptoms. On the contrary, greater adherence to sweets and traditional Tianjin dietary patterns was associated with a higher prevalence of depressive symptoms.
In order to reveal the quantitative relationship between fatigue crack deflection path and cross-sectional grain boundary (GB) arrangement of metallic nanolayered composites (NLCs), a stochastic model was established based on the interface-dominant fatigue damage for the ultrafine-scale NLCs. The model indicates that the crack deflection length decreases with decreasing GB arrangement deviation and grain size of constituent layers. The observation and quantitative analysis of fatigue cracking behavior of the Cu/W multilayers with a layer thickness of 5 and 20 nm was conducted to verify the model.
Dalian, China, is a city free of rabies in recent 20 years, but the annual cost for rabies vaccination still brings an economic burden on society and individuals. We did a retrospective descriptive analysis to analyse the reason for this and try to find some ways to resolve it. A total of 10 028 post-exposure prophylaxis (PEP) cases were recorded from January 2016 to December 2017. According to the exposure grades, 32 cases were grade I; 7712 cases were grade II; 2284 cases were grade III. All the patients in the cases were injured by pet dogs without abnormal clinical signs, and 80% of them were home pet dogs. Fifty-two per cent of the pet dogs were vaccinated. All the dogs survived during the PEP vaccination period. The data showed that a considerable proportion of people who did not have exposure risk for rabies had received vaccination. The underlying reasons included social, medical and personal factors. So here we proposed to replace the current ‘five-course’ intramuscular injection with intradermal injection method in the cities free of rabies in China, this can not only achieve effective vaccination but also save resources and eliminate the fear of rabies from victims. Meanwhile we should strengthen communication on rabies knowledge and make a routine evaluation of rabies surveillance system to improve understanding of the risk for rabies from biting animals.
Subacute ruminal acidosis (SARA) can increase the level of inflammation and induce rumenitis in dairy cows. Berberine (BBR) is the major active component of Rhizoma Coptidis, which is a type of Chinese anti-inflammatory drug for gastrointestinal diseases. The purpose of this study was to investigate the anti-inflammatory effects of BBR on lipopolysaccharide (LPS)-stimulated rumen epithelial cells (REC) and the underlying molecular mechanisms. REC were cultured and stimulated with LPS in the presence or absence of different concentrations of BBR. The results showed that cell viability was not affected by BBR. Moreover, BBR markedly decreased the concentrations and mRNA expression of pro-inflammatory cytokines, including tumor necrosis factor-α, interleukin-1β, and interleukin-6 in the LPS-treated REC in a dose-dependent manner. Importantly, Western blotting analysis showed that BBR significantly suppressed the protein expression of toll-like receptor 4 (TLR4) and myeloid differentiation primary response protein (MyD88) and the phosphorylation of nuclear factor-κB (NF-κB), inhibitory kappa B (IκBα), p38 mitogen-activated protein kinase (MAPK), and c-Jun N-terminal kinase (JNK) in LPS-treated REC. Furthermore, the results of immunocytofluorescence showed that BBR significantly inhibited the nuclear translocation of NF-κB p65 induced by LPS treatment. In conclusion, the protective effects of BBR on LPS-induced inflammatory responses in REC may be due to its ability to suppress the TLR4-mediated NF-κB and MAPK signaling pathways. These findings suggest that BBR can be used as an anti-inflammatory drug to treat inflammation induced by SARA.
Nanocrystalline and nanolaminated materials show enhanced radiation tolerance compared with their coarse-grained counterparts, since grain boundaries and layer interfaces act as effective defect sinks. Although the effects of layer interface and layer thickness on radiation tolerance of crystalline nanolaminates have been systematically studied, radiation response of crystalline/amorphous nanolaminates is rarely investigated. In this study, we show that irradiation can lead to formation of nanocrystals and nanotwins in amorphous CuNb layers in Cu/amorphous-CuNb nanolaminates. Substantial element segregation is observed in amorphous CuNb layers after irradiation. In Cu layers, both stationary and migrating grain boundaries effectively interact with defects. Furthermore, there is a clear size effect on irradiation-induced crystallization and grain coarsening. In situ studies also show that crystalline/amorphous interfaces can effectively absorb defects without drastic microstructural change, and defect absorption by grain boundary and crystalline/amorphous interface is compared and discussed. Our results show that tailoring layer thickness can enhance radiation tolerance of crystalline/amorphous nanolaminates and can provide insights for constructing crystalline/amorphous nanolaminates under radiation environment.
This study aimed to improve the performance of an intermediate covering of soils in landfill sites by using agents such as calcined lime, sawdust and polyacrylamide (PAM). Compressive strength, permeability and water-holding capacity of modified soils were measured, and the effects of regulating pH and pollutant removal in leachate were also investigated in a leaching experiment. The results indicate that three modifying agents could improve the compressive strength of an intermediate soil cover. The permeability of lime-treated soil increased as the amount of lime increased, while that of sawdust- and PAM-modified soil declined. Results from a leaching experiment show that lime- and sawdust-modified soils could improve leachate quality. The pH value of leachate from 5% lime-modified soils was 7.78, which is suitable for the metabolism of anaerobic microorganisms. The removal efficiencies of chemical oxygen demand, total organic carbon, total nitrogen and volatile fatty acids in leachate permeating lime- and sawdust-modified intermediate cover was improved so that the pollution load of leachate was reduced. The water-holding capacities for 20% sawdust and 0.5% PAM-modified soils were 65.19% and 43.52%, respectively, which helps to maintain the optimum water content of landfill. The water-holding capacity of PAM-modified samples declined in alkaline soil. It is concluded that the combination of 5% sawdust, 5% lime and 90% soil would be optimal for an intermediate covering layer.
Dairy cows with fatty liver or ketosis display decreased insulin sensitivity and defects in the insulin receptor substrate (IRS)/PI3K/AKT signaling pathway. Phosphatase and tensin homolog (PTEN) is a well-known tumor suppressor and also a negative regulator of insulin signaling and peripheral insulin sensitivity. We investigated the hypothesis that PTEN may affect the insulin pathway-mediated hepatic glucose and lipid metabolism in dairy cows. Adenovirus vectors that over-express and silence PTEN were constructed, and then transfected into hepatocytes isolated from calves to investigate the effect of PTEN on PI3K/AKT signaling pathway. PTEN silencing increased the phosphorylation of AKT and the expression of PI3K but decreased the phosphorylation of IRS1, which increased the phosphorylation levels of glycogen synthase kinase-3β (GSK-3β) and expression of sterol regulatory element-binding protein-1c (SREBP-1c). Increased GSK-3β phosphorylation further up-regulated expression of the key enzymes phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6-Pase) involved in gluconeogenesis. Furthermore, the expression of SREBP-1c target gene fatty acid synthase (FAS) also increased significantly. We further showed that PTEN over-expression could reverse the above results. PTEN negatively regulates the enzymes involved in hepatic gluconeogenesis and lipid synthesis, which suggests that PTEN may be a therapeutic target for ketosis and fatty liver in dairy cows.
Seed reserves play vital roles in seed germination and seedling growth and their variation may be related to various environment factors, plant traits and phylogenetic history. Here, the evolutionary correlation associated with seed mass and altitude and carbon (C), nitrogen (N) and phosphorus (P) allocation of seeds among 253 alpine herbaceous plants was tested. In this study, phylogeny had strong limitations on nutrient allocation of seeds across species, and species from younger phylogenetic groups tended to have higher N and P contents, which might be considered as the evolutionary selection of seed plants. Higher seed N and P content would help seedlings to gain more survival chance and stronger competitive capacity, and their progeny would be more likely to be preserved. When phylogeny was considered, altitude only had a significant positive effect on P content, but the negative effects on seed mass were all expressed. The independent effects of altitude and seed mass suggest that the nutrient allocation of seeds might be affected by both environment and plant traits. In addition, altitude and seed mass displayed partial overlapping effects on nutrient allocation of seeds. The negative effects of seed mass were affected slightly by altitude, whereas altitude only had a significant positive effect on P content when seed mass was controlled. Above all, seed P content showed obvious and general correlations with seed mass, altitude and age of clade, which indicated that higher seed P content might be an adaptive selection of species associated with growth and survival of progeny.