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High-throughput western blot (WB) analysis of small and precious samples, such as various age-related subtype-specific human induced neurons (hiNs), confers the ability to obtain more consistent, comparable, and informative data from materials with extremely limited availability. In this study, p-toluenesulphonic acid (PTSA), an odorless tissue fixative, was used to inactivate HRP for developing a high-throughput WB method. PTSA-treated blots showed fast and efficient inactivation of HRP without detectable protein loss and epitope damage. With a brief PTSA-treatment before every next probing, 10 proteins of dopaminergic hiNs could be sequentially, sensitively, and specifically detected in a blot. These WB data proved the age-associated and neuron-specific features of hiNs and further revealed a sharp reduction of two Parkinson’s disease-associated proteins, UCHL1 and GAP43, in the normal aging dopaminergic neurons. Together, this study developed a unique and high-efficiency WB analysis and pinpointed its special value for capturing robust useful data from limited, precious samples.GRAPHICAL ABSTRACTHIGHLIGHTS✓P-toluenesulphonic acid (PTSA) quickly and fully deactivated HRP on immunoblots.✓PTSA was an odorless, non-volatile, low cost, and user-friendly HRP inactivator.✓PTSA allowed high-efficiency WB analysis to save small precious samples and time.✓10 proteins were detected in a single blot of age-relevant human induced neurons.✓UCHL1 and GAP43 sharp decline occurred in aging human induced dopaminergic neurons.

In this work, two Cu(II)-containing coordination polymers, namely, {[Cu2(L)2(dpyb)(H2O)4]·6H2O}n (1) and {[Cu2(L)2(dpyb)](CH3CN)2}n (2) have been hydrothermally synthesized based on a substituted t...

The goal of the present work is to investigate the effects of varying coatings on in vitro mineralization of silk fibroin fiber artificial ligaments (SFALs). Silk fibroin (SF) and sodium alginate (SA) aqueous solutions with varying concentrations were respectively employed for coating surfaces of the SFALs to improve calcium phosphate (CaP) deposition. Scanning electron microscopy, Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy and Energy-dispersive X-ray spectroscopy were utilized for characterizations. Furthermore, the surface of the SFAL was modified with SF/SA composite coatings. The volume ratios of the SF to the SA were set at 2:8, 4:6, 5:5, 6:4 and 8:2. The results show that either the SF or the SA coatings can improve the mineralization of the SFALs in vitro. The concentration of 1% for the SF and the SA coatings is conducive to deposit more calcium ions and obtain a biomimetic Ca/P ratio. Furthermore, the SF/SA composite coatings facilitate a thicker and uniform deposition layer than the SF and the SA coatings. The sediments are spherical particles and are more uniform in size. Therefore, the SF/SA composite coatings have more significant efficacy, particularly when the volume ratio of the SF to the SA was 4:6. The present work indicates that the SF/SA coatings possess excellent potential in promoting the osseointegration of artificial ligaments implanted in bone tunnels.

This study aimed to evaluate the expression level of caseins during mastitis of goats. Whole goat caseins were used as primary antigens for mouse immunization. A monoclonal antibody (mAb) named 5B with high specificity to goat αs1-casein was developed. Further results showed that mAb 5B can successfully be applied to western blot and ELISA. In addition, immunofluorescence analysis using this mAb showed increased milk αs1-casein level in mastitis-affected goats. In conclusion, this study has established an effective tool to evaluate the expression level of αs1-casein during mastitis development.

In this paper, the numerical study focuses on the stress field of the knee meniscus in the two gaits of the jumping motion, and the knee injury and its knee protection effect are studied based on the stress field distribution characteristics of the knee meniscus. The jumping gait simulation was carried out by using a more detailed healthy knee model including bone, articular cartilage, meniscus, ligaments and peripheral soft tissues constructed by combining CT and MRI tomography, and the peak stress and its distribution area were analyzed based on the stress field characteristics of the meniscus, and the knee meniscus and its injury and protection under different knee pads were discussed. The results showed that the anterior angle of the meniscus on the medial knee joint was an area prone to injury in the take-off and landing gait. Under the knee pads, the peak stress of the meniscus decreases, and its distribution area gradually shifts from the anterior angle to the middle, and the functional knee pads with the optimized force load significantly alleviate the stress concentration of the meniscus. Therefore, the numerical study of the stress field of the knee meniscus provides theoretical support and optimization guidance for the design of functional knee pads.

Spinocerebellar ataxias (SCAs) are a large group of hereditary neurodegenerative diseases characterized by ataxia and dysarthria. Due to high clinical and genetic heterogeneity, many SCA families are undiagnosed. Herein, using linkage analysis, WES, and RP-PCR, we identified the largest SCA36 pedigree in Asia. This pedigree showed some distinct clinical characteristics. Cognitive impairment and gaze palsy are common and severe in SCA36 patients, especially long-course patients. Although no patients complained of hearing loss, most of them presented with hearing impairment in objective auxiliary examination. Voxel-based morphometry (VBM) demonstrated a reduction of volumes in cerebellum, brainstem, and thalamus (corrected P 0.05). Reduced volumes in cerebellum were also found in presymptomatic carriers. Resting-state functional MRI (R-fMRI) found reduced ReHo values in left cerebellar posterior lobule (corrected P 0.05). Diffusion tensor imaging (DTI) demonstrated a reduction of FA values in cerebellum, midbrain, superior and inferior cerebellar peduncle (corrected P 0.05). MRS found reduced NAA/Cr values in cerebellar vermis and hemisphere (corrected P 0.05). Our findings could provide new insights into management of SCA36 patients. Detailed auxiliary examination are recommended to assess hearing or peripheral nerve impairment, and we should pay more attention to eye movement and cognitive changes in patients. Furthermore, for the first time, our multimodel neuroimaging evaluation generate a full perspective of brain function and structure in SCA36 patients.

Ethnopharmacological relevance Sugemule-3 decoction (SD-3) is a commonly used prescription in Mongolian medicine which composed of the herbs Baidoukou (the fruit of Amomum compactum Sol. ex Maton), Baijusheng (the fruit of Lactuca sativa L.) and Biba (Piper longum L.). SD-3 has remarkable effect on the cardiovascular diseases, but its pharmacological mechanism has not been elucidated. Aim of this study To evaluate the cardioprotective effects and the potential mechanisms of the ethanol extracts of SD-3 against isoproterenol (ISO)-induced heart failure (HF) in rats. Material and methods The ethanol extracts of SD-3 were prepared and analyzed by LC-ESI-MS/MS. One hundred male Wistar rats were randomly divided into five groups: control, ISO (HF) and different doses of SD-3 (0.4, 0.2, 0.1 g/kg/d) groups. HF model rats were established by intraperitoneal injecting of ISO. The left ventricular function was evaluated by echocardiography. Myocardial injury and fibrosis were examined by hematoxylin-eosin (HE) and Masson staining. Western-blot analysis was performed to determine the protein expression of apoptosis and mitochondrial dynamics in all the groups. Moreover, the structural changes in the mitochondria of cardiomyocytes were also observed by transmission electron microscopy. Results Fifteen compounds were detected in the ethanol extracts of SD-3, include piperine, piperanine, etc. Rats administered with ISO showed a significant decline in the left ventricular function. The cardiac histopathological changes such as local necrosis, interstitial oedema, and cardiac fibrosis were also observed in the ISO group. The treatment with SD-3 significantly inhibited these effects of ISO. ISO was found to increase the protein expression of Bax, cleaved-PARP and cleaved-caspase-3, -7 -9, destroy the balance between mitochondrial fusion and fission, and alter the mitochondrial morphology. The ethanol extracts of SD-3 could rebalance mitochondrial fusion and fission, and ameliorates the morphological abnormalities induced by ISO in mitochondria. Conclusion The current study demonstrated that ethanol extracts of SD-3 improved isoprenaline-induced cardiac hypertrophy and fibrosis through inhibiting cardiomyocyte apoptosis and regulating the mitochondrial dynamics.

BackgroundGoat milk is essential for the initial development of kids by providing a great source of commensal bacteria. Here we analyzed the microbiota of goat colostrum which was collected daily for five days post delivery and mature milk collected at the 7th, 10th, 20nd, 30th, 40th, 50th and 60th days, respectively, from three farms of Shaanxi province.ResultsThe result showed that microbial alpha diversity was higher in the mature milk compared with that in the colostrum. According to taxonomy results, Proteobacteria, Firmicutes, Actinobacteria and Bacteroidetes were the predominant bacteria phyla in both colostrum and mature milk. In addition, lactation stage noticeably influenced the composition of milk microbiota. Specifically, Novosphingobium, Brachybacterium, Psychrobacter, Lactobacillus, Yersinia, Roseatelesand, Rothia, Sanguibacter, Cloacibacterium, Variovorax, Sphingobacterium, Coxiella were enriched in the colostrum while Georgenia, Peptostreptococcus, Bacteroidales, Yaniella, Planomicrobium, Cloacibacterium, Azospirillum, Turicibacter, Cupriavidus, Herbaspirillum, Rhodobacteraceae, Aeromonadales were the dominant genera in the mature milk. The enriched metabolic functions of the goat milk microbiota were predicted by PICRUSt and classified by KEGG patway. Moreover, the abundances of the environmental information processing, cellular processes pathway, genetic information processing pathway, organismal systems pathway and metabolism pathway were significantly different between microbiota of colostrum and mature milk.ConclusionsAltogether, our study disclosed the significant difference between the microbial communities of colostrum and mature milk and provided grounds for further research in dairy microbiology.

Porous graphene-like carbon materials (PGCMs) for supercapacitors were synthesized from petroleum pitch by a nano-CaCO3 template strategy combined with KOH activation. The PGCMs were characterized by TEM, XPS, Raman spectroscopy and N2 adsorption. Results show that their specific surface areas are 1542-2305 m2·g−1, depending on the template/pitch ratio, KOH/pitch ratio and activation temperature. The PGCMs feature interconnected graphene-like carbon layers with abundant small pores. The optimum supercapacitor electrode is produced when the PGCM is synthesized with a template/pitch ratio of 1.5, a KOH/pitch ratio of 1.5 at 850 °C for 1h. It has a specific capacitance of 293 F·g−1 at a current density of 0.05 A·g−1, an excellent rate capability with a capacitance of 231 F·g−1 at a current density of 20 A·g−1 and an outstanding cycling stability with a 97.4% capacitance retention after 7000 cycles in a 6 M KOH aqueous electrolyte. It also exhibits a high specific capacitance of 267 F·g−1 at a current density of 0.05 A·g−1, and a high energy density of 148.3 Wh·kg−1 at a power density of 204.2 W·kg−1 in a BMIMPF6 ionic liquid electrolyte. This is a possible method for the synthesis of PGCMs for high-performance supercapacitors.

This paper presents an application of ultra high-performance liquid-chromatography-quadrupole-TOF high resolution mass spectrometry (UHPLC-Q-TOF HRMS) for simultaneous analysis of 23 illegal adulterated aphrodisiac type chemical ingredients in health foods and Chinese Traditional Patent Medicines (CTPMs). The mass spectrometer was operated in Information Dependent Acquisition (IDA) mode, which provides crucial information for the elemental composition analysis, structure elucidation and quantitative analysis simultaneously. Quantitative analysis was performed using the peak areas of the precursor ions in the XICs. The method validation included assessment of selectivity, sensitivity, calibration curve, accuracy, precision, recovery, matrix effect and stability. The results show good linear relationship with the concentrations of the analytes over wide concentration ranges (e.g., 0.05–10 μg/g for sildenafil) as all the fitting coefficients of determination r2 are >0.9984. The detection limits (LODs) were in the range of 0.002–0.1 μg/g. The recoveries were able to reach 82.5–103.6%, while the matrix effects ranged from 87.7 to 109.3%. The intra- and inter-day accuracies were in the range of 82.3–113.8%, while the intra- and inter-day precision ranged from 0.4 to 13.6%. Among 40 batches of health foods and 32 batches of CTPMs (including 28 capsules, 32 tablets, 10 liquid and 2 pills) samples, 28 batches of heath foods were positive. The detected chemical ingredients involved sildenafil, tadalafil, aildenafil and sulfoaildenafil. This method can be used for the screening, identification and quantification of illegal adulterated aphrodisiac chemical ingredients in health foods and CTPMs. Moreover, the LC-Q-TOF MS is very useful to structural elucidation of unknown compound. Keywords: Q-TOF MS, Illegal adulteration, Aphrodisiac chemical ingredients, Qualitation, Quantitation

Contagious pustular dermatitis is an exanthematous zoonotic disease caused by the orf virus. Pandemic outbreaks of this disease cause great economic losses, while the pathogenesis of this disease still remains obscure. In this study, blood samples were collected from 628 asymptomatic goats across China for PCR-based virus detection. We detected the orf virus in the blood of asymptomatic goats. Moreover, the orf virus obtained from the blood of infected goats was infectious and induced typical symptoms of contagious pustular dermatitis after inoculation of uninfected dairy goats. In summary, our data provide evidence that asymptomatic animals may be carriers of orf virus. Our findings should contribute to elucidating the details underlying the pathogenesis of contagious pustular dermatitis.

For the past few years, AI and deep-learning got a second wind in the development of Brain neuroscience and Big data. In this paper, we describe an analog neuron model by using current-steering DAC circuit which uses analog circuits simulate the internal architecture of the neuron: Dendrites, axon, synapses and soma. The architecture we propose can achieve most of the network architecture, and the expandable neuron architecture that the hardware architecture can be adjusted according to the size of the system for better energy efficiency.

Contagious caprine pleuropneumonia (CCPP) is a highly contagious infectious disease of goats caused by Mycoplasma capricolum subspecies capripneumoniae (Mccp). CCPP outbreaks usually result in high morbidity and mortality of the affected goats, making this disease a major cause of economic losses to goat producers globally. However, the pathogenesis of CCPP remains unclear. Here, we show that IL-17-driven neutrophil accumulation is involved in the lung damage in CCPP goats. During CCPP development, intense inflammatory infiltrates could be observed in the injured lungs. Specifically, neutrophils were observed to be present within the alveoli. Increased IL-17 release drove the excessive influx of neutrophils into the lung, as IL-17 effectively stimulated the production of neutrophil chemoattractants from lung epithelial cells following Mccp infection. Our data highlight a critical role of IL-17-driven neutrophil accumulation in the pathogenesis of CCPP and suggest that IL-17 may potentially be a useful immunotherapeutic target for the treatment of CCPP.

The evolution of organisms has provided a variety of mechanisms to maintain the integrity of its genome, but as damage occurs, DNA damage repair pathways are necessary to resolve errors. Among them, the DNA double-strand break repair pathway is highly conserved in eukaryotes, including mammals. Nonhomologous DNA end joining and homologous directed repair are two major DNA repair pathways that are synergistic or antagonistic. Clustered regularly interspaced short palindromic repeats genome editing techniques based on the nonhomologous DNA end joining repair pathway have been used to generate highly efficient insertions or deletions of variable-sized genes but are error-prone and inaccurate. By combining the homology-directed repair pathway with clustered regularly interspaced short palindromic repeats cleavage, more precise genome editing via insertion or deletion of the desired fragment can be performed. However, homologous directed repair is not efficient and needs further improvement. Here, we describe several ways to improve the efficiency of homologous directed repair by regulating the cell cycle, expressing key proteins involved in homologous recombination and selecting appropriate donor DNA.

Clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 (Cas9) is a precise genome manipulating technology that can be programmed to induce double-strand break (DSB) in the genome wherever needed. After nuclease cleavage, DSBs can be repaired by non-homologous end joining (NHEJ) or homology-directed repair (HDR) pathway. For producing targeted gene knock-in or other specific mutations, DSBs should be repaired by the HDR pathway. While NHEJ can cause various length insertions/deletion mutations (indels), which can lead the targeted gene to lose its function by shifting the open reading frame (ORF). Furthermore, HDR has low efficiency compared with the NHEJ pathway. In order to modify the gene precisely, numerous methods arose by inhibiting NHEJ or enhancing HDR, such as chemical modulation, synchronized expression, and overlapping homology arm. Here we focus on the efficiency and other considerations of these methodologies.

Objective: Atherosclerosis (AS) is an inflammatory disease and the formation of atherosclerotic plaque plays a critical role in AS progression. We aimed to investigate the effect of long non-coding RNA (lncRNA) activated by DNA damage (NORAD)/microRNA-495-3p (miR-495-3p)/Kruppel-like factor 5 (KLF5) axis on atherosclerotic plaque formation. Methods: The ApoE−/- mice were fed a high-fat diet to construct AS mouse models and the modeled mice were treated with altered NORAD, miR-495-3p or KLF5. NORAD, miR-495-3p and KLF5 expression in mouse aorta tissues were evaluated, and the levels of inflammatory factors, oxidative stress factors, endothelial function indices and blood lipid in mice were all determined. The atherosclerotic plaque area, lipid deposition area, collagen fibers and CD68 expression in mouse aorta tissues were assessed. The regulatory relation between NORAD and miR-495-3p, and the target relation between miR-495-3p and KLF5 were confirmed. Results: NORAD and KLF5 were increased whereas miR-495-3p was decreased in atherosclerotic mouse aortas. Inhibited NORAD or elevated miR-495-3p suppressed inflammation, oxidative stress, endothelial dysfunction, blood lipid level, atherosclerotic plaque area, collagen fibers and CD68 expression in atherosclerotic mouse aortas. Effects of elevated miR-495-3p on atherosclerotic mice could be reversed by up-regulation of KLF5. NORAD served as a sponge of miR-495-3p and miR-495-3p directly targeted KLF5. Conclusion: Silenced NORAD elevated miR-495-3p to suppress atherosclerotic plaque formation via reducing KLF5. Findings in our research may be helpful for exploring molecular mechanisms of AS.

Flixweed (Descurainia Sophia L.) is a problematic weed in winter wheat fields in China, which causes great loss of wheat yield. A total of 46 flixweed accessions from winter wheat-planting areas were collected and used for the survey of resistance to tribenuron-methyl and Pro197 mutation diversity. According to the “R” resistance rating system, 16 flixweed accessions have evolved resistance to tribenuron-methyl, 13 accessions have high risk of developing resistance to this herbicide and 17 accessions are susceptible. The mutation of Pro197 codon (CCT) changed proline (Pro) into leucine (Leu) (homozygous, RR), serine (Ser, RR), histidine (His, RR), threonine (Thr, RR), Pro/Leu (heterozygous, RS), Pro/Ser (RS), Pro/His, Pro/Thr (RS) and Pro/Tyr (RS). Among these amino acid changes, a Pro197-Pro/Tyr (heterozygous, RS) substitution caused by the mutation of two successive nucleotides was identified for the first time in resistant weed species. In addition, the Pro197-His and Pro197-Pro/His mutations have not been reported previously in flixweed. Finally, a CPAS marker was developed to identify flixweed plants with or without Pro197 mutation.

Soybean [Glycine max (L.) Merr.] is a widely cultivated crop, whose growth and development are usually influenced by fluctuating light environment in fields. In the present study, two soybean cultivars (Nandou12 and Nan 032-4) were selected to study the regulatory mechanisms of capability to do photosynthetic work under different light intensities. Results showed that the net photosynthetic rate (P n ) of Nandou 12, which has a higher yield in field condition, was significantly higher than that of Nan 032-4 under all the light intensities studied. Measurements on stomatal conductance (G s) and intercellular CO2 concentration (C i) suggested that the difference of P n between these two cultivars was mainly caused by non-stomatal factors. Transcript detection found that the expression of genes encoding plastid terminal oxidase (PTOX) and alternative oxidase (AOX), which represent the terminal oxidases of chlororespiration in chloroplasts and alternative respiration in mitochondria, respectively, was up-regulated in both cultivars when light intensity increased, implying potential roles of these two terminal oxidases in photosynthetic regulation. Further analysis showed that the expression level of PTOX increased more obviously in Nandou 12, together with higher non-photochemical quenching (NPQ) and higher activities of superoxide dismutase (SOD) and ascorbate peroxidase (APX) than that in Nan 032-4, when light intensity increased. However, Nan 032-4 performed a more obvious up-regulation of AOX genes, as well as higher activities of NAD-malate dehydrogenase (MDH) and NADP-MDH than Nandou 12, when light intensity increased. These distinctions were consequently suggested to be potential reasons for the difference of photosynthetic performance between two soybean cultivars.

The block cipher used in the Chinese Wireless LAN Standard (WAPI), SMS4, was recently renamed as SM4, and became the block cipher standard issued by the Chinese government. This paper gives a method for finding the linear approximations of SMS4. With this method, 19-round one-dimensional approximations are given, which are used to improve the previous linear cryptanalysis of SMS4. The 19-round approximations hold with bias 2−62.27; we use one of them to leverage a linear attack on 23-round SMS4. Our attack improves the previous 23-round attacks by reducing the time complexity. Furthermore, the data complexity of our attack is further improved by the multidimensional linear approach.

The aim of this study is to investigate whether exogenous application of salicylic acid (SA) could modulate the photosynthetic capacity of soybean seedlings in water stress tolerance, and to clarify the potential functions of terminal oxidase (plastid terminal oxidase (PTOX) and alternative oxidase (AOX)) in SA′ s regulation on photosynthesis. The effects of SA and water stress on gas exchange, pigment contents, chlorophyll fluorescence, enzymes (guaiacol peroxidase (POD; EC 1.11.1.7), superoxide dismutase (SOD; EC 1.15.1.1), catalase (CAT; EC 1.11.1.6), ascorbate peroxidase (APX; EC 1.11.1.11) and NADP-malate dehydrogenase (NADP-MDH; EC1.1.1.82)) activity and transcript levels of PTOX , AOX1 , AOX2a , AOX2b were examined in a hydroponic cultivation system. Results indicate that water stress significantly decreased the photosynthetic rate (Pn), stomatal conductance (Gs), transpiration rate (E), pigment contents (Chla + b, Chla/b, Car), maximum quantum yield of PSⅡphotochemistry (Fv/Fm), efficiency of excitation capture of open PSⅡcenter (Fv’/Fm’), quantum efficiency of PSⅡphotochemistry (ΦPSⅡ), photochemical quenching (qP), and increased malondialdehyde (MDA) content and the activity of all the enzymes. SA pretreatment led to significant decreases in Ci and MDA content, and increases in Pn, Gs, E, pigment contents, Fv/Fm, Fv’/Fm’, ΦPSⅡ, qP, and the activity of all the enzymes. SA treatment and water stress alone significantly up-regulated the expression of PTOX , AOX1 and AOX2b . SA pretreatment further increased the transcript levels of PTOX and AOX2b of soybean seedling under water stress. These results indicate that SA application alleviates the water stress-induced decrease in photosynthesis may mainly through maintaining a lower reactive oxygen species (ROS) level, a greater PSⅡefficiency, and an enhanced alternative respiration and chlororespiration. PTOX and AOX may play important roles in SA-mediated resistance to water stress.

In this paper, according to the traction calculation, a stray current dynamic distribution model of multi-locomotives running under different operation conditions was established which based on the metro locomotives stray current static distribution model, then analyzed the dynamic distribution of rail potential and stray current under different operation conditions. The simulation results show that: locomotives operation conditions have a significant influence on the distribution of rail potential and stray current. The more locomotives running, the greater value and stronger fluctuation of stray current.

A series of six full-scale cruciform H-shaped beam to square tube column joint with cast steel stiffener specimens was tested under cyclic loading to study the seismic behavior of the new joints. A parametric numerical analysis based on the test results was also conducted. The hysteretic curves of specimens not filled with concrete were full, and the ones of specimens filled with concrete were pinched. The maximum moment resistance of the joints filled with concrete in the column increased from 10 to 30% compared with that of the joints not filled with concrete. The energy dissipation coefficient of the joints not filled with concrete in the column varied from 2.681 to 3.585, whereas it changed from 1.892 to 1.711 for the joints filled with concrete when the width of the square tube column ranged from 300 to 500 mm. The results demonstrate that the joints with cast steel stiffeners have excellent moment capacity, deformation performance, and energy dissipation coefficients. The new joints could po...

This paper presents a design and its implementation of our automatic vehicle that travels outdoors without drivers. Users program the routes by our proposed smartphone application which routes are from Google map API. The vehicle applies the sensors of GPS, LiDAR, and cameras to direct the driving path. The sonars are also applied to the collision prevention in short distance. Based on the restriction of traffic rules, the proposed vehicle be used in close area transportation, such as the campuses and factories.

In order to assess the protective effects of the DNA vaccine (pcDip/pilE) against Legionella pneumophila , the coding sequences of the 2 proteins were cloned into the pET32a(+) and pcDNA3.1(+) vectors. To provide an enhanced immunological response, the proteins were linked together. In this study, the A/J mouse model was used for examine the immunogenicity and protective efficacy of the DNA vaccines of pcDip, pcDpilE and pcDip/pilE. Our results showed that the total IgG titers were higher level increasing after the stimulation of pcDip/pilE than pcDip and pcDpilE immunization. The DNA vaccine (pcDip/pilE) can protect the A/J mouse against a higher dose (2 × 10 7 L. pneumophila cells) of L. pneumonia compared to the other single-DNA vaccine in our study, and the ratio of the survival reached 100% in 10 days after the last DNA vaccine immunization. Our study indicates that these findings provide experimental evidence to support the claim that pcDip/pilE may be an efficient DNA vaccine against Legionella pneumophila .

Hard target penetration acceleration signals are non-stationary signals. In this paper use the local mean decomposition theory to analysis hard target penetration acceleration signals. Local mean decomposition can adaptively decompose any complex non-stationary signal into a number of physically meaningful instantaneous frequency components and these components could well reflect the intrinsic nature of the signals. Use local mean decomposition to separate the rigid projectile overload from hard target penetration acceleration signals. That provide reliable data to support accurately calculating the effective penetration depth, hard floors or holes through the number of target parameters on the penetration of weapons is significant. The author apply local mean decomposition algorithm in penetration acceleration signal separation and the results verify the effectiveness of the method.

Zinc is an essential element that facilitates coordination of immune activation during the host response to infection. We recently reported that zinc deficiency increases systemic inflammation, vital organ damage, and mortality in a small animal model of sepsis. To investigate potential mechanisms that cause these phenomena, we used the same animal model and observed that zinc deficiency increases bacterial burden and enhances NF-kappaB activity in vital organs including the lung. We conducted further studies in the lung to determine the overall impact of zinc deficiency. At the molecular level, NF-kappaB p65 DNA-binding activity was enhanced by zinc deficiency in response to polymicrobial sepsis. Furthermore, expression of the NF-kappaB-targeted genes IL-1beta, TNFalpha, ICAM-1, and the acute phase response gene SAA1/2 were elevated by zinc deficiency. Unexpectedly, the amount of NF-kappaB p65 mRNA and protein was increased in the lung including alveolar epithelia of zinc-deficient mice. These events occurred with a significant and concomitant increase in caspase-3 activity within 24 h of sepsis onset in zinc-deficient mice relative to control group. Short-term zinc supplementation reversed these effects. Reconstitution of zinc deficiency in lung epithelial cultures resulted in similar findings in response to TNFalpha. Taken together, zinc deficiency systemically enhances the spread of infection and NF-kappaB activation in vivo in response to polymicrobial sepsis, leading to enhanced inflammation, lung injury, and, as reported previously, mortality. Zinc supplementation immediately before initiation of sepsis reversed these effects thereby supporting the plausibility of future studies that explore zinc supplementation strategies to prevent sepsis-mediated morbidity and mortality.

In the paper, the notions of generalized loose (weak) saddle points for set-valued mappings are introduced. By applying the fixed point theorem and the scalarization method, some new existence theorems for generalized loose (weak) saddle points are established in locally G -convex space. As applications, some vector minimax theorems are given also.

The cellular generation of toxic metabolites and subsequent detoxification failure can cause the uncontrolled accumulation of these metabolites in cells, leading to cellular dysfunction. Amyloid-beta protein (A beta), a normal metabolite of neurons, tends to form toxic oligomeric structures that cause neurodegeneration. It is unclear how healthy neurons control the levels of intracellular oligomeric A beta in order to avoid neurodegeneration. Using immunochemical and biochemical studies, we show that the A beta-binding serine protease Omi is a stress-relieving heat-shock protein that protects neurons against neurotoxic oligomeric A beta. Through its PDZ domain, Omi binds preferentially to neurotoxic oligomeric forms of A beta rather than non-toxic monomeric forms to detoxify oligomeric A beta by disaggregation. This specific interaction leads not only to mutual detoxification of the pro-apoptotic activity of Omi and A beta-induced neurotoxicity, but also to a reduction of neurotoxic-A beta accumulation. The neuroprotective role of Omi is further supported by its upregulation during normal neurogenesis and neuronal maturation in mice, which could be in response to the increase in the generation of oligomeric A beta during these processes. These findings provide novel and important insights into the detoxification pathway of intraneuronal oligomeric A beta in mammals and the protective roles of Omi in neurodegeneration, suggesting a novel therapeutic target in neurodegenerative diseases.

Bamboo charcoal supporting silver (BC/Ag) was prepared by activation and chemical reduction. The BC/Ag composites were characterized by silver particle size and distribution, silver ion (Ag + ) release and antibacterial properties. Scanning and transmission electron microscopy (SEM and TEM) showed that the Ag particles were distributed uniformly on the BC matrix. The Ag particle size was found to be less than 150 nm based on TEM. The Ag content and surface morphology of the BC/Ag composites depended on the initial concentration of AgNO 3 , and the higher the Ag content, the smaller the specific surface area obtained on the BC. The antibacterial effects of the BC/Ag composite powders were assessed from the minimum inhibitory concentrations (MICs) and by the plate-counting method, and an excellent antibacterial performance was discovered.

Diosgenyl saponins are the most abundant steroid saponins, and exert a large variety of biological functions. In a previous report, we showed that dioscin was able to induce cytotoxicity and apoptosis in human myeloblast leukemia HL-60 cells. This study further investigated the action mechanisms underlying this effect. The activation of caspase-9 and -3, but not caspase-8, together with the down-regulation of anti-apoptotic Bcl-2 protein, demonstrated that the apoptotic signaling triggered by dioscin was mediated through the intrinsic mitochondria-dependent pathway. We also investigated its anti-proliferative effect on human chronic myelogenous leukemia K562 cells. Flow cytometry analysis showed that dioscin treatment induced the accumulation of cells in the G(2)/M phase. Cytomorphology with DAPI and Wright-Giemsa staining demonstrated the enlargement of cell volume and multinucleation in the treated cells. Subsequent apoptosis was delineated with phosphatidylserine externalization and DNA hypodiploidy. Trillin was one of the hydrolysates of dioscin. We demonstrated that it could induce multinucleation in HL-60, K562 and human promyelocytic leukemia NB(4) cells, suggesting its extensive mitotic-arresting effects. As the diosgenyl sapogenin, diosgenin was also shown to be able to induce multinucleation and apoptosis in K562 cells in a similar manner to dioscin. These findings suggest that diosgenyl saponins have the properties to induce mitotic arrest and apoptosis, suggesting that they may be a new kind of antimitotic agent.

The transcription factor CHOP/GADD153 is reportedly induced by cellular stresses such as UV light, genotoxic agents, and protein misfolding in the endoplasmic reticulum. However, the mechanism whereby induction of the GADD153 gene is linked to a downstream pathway is still unclear. Previously, we observed that a synthetic retinoid N-(4-hydroxyphenyl)retinamide (4HPR) effectively impaired cell growth and survival (induction of growth arrest and apoptosis) in human hepatoma cells, which was accompanied by over expression of GADD153. Furthermore, GADD153-transfected Hep 3B cells were growth arrested and were sensitized to drug-induced apoptosis. Thus, in this study, we used suppression subtractive hybridization (SSH) to identify GADD153 target genes that were up-regulated or down-regulated in the GADD153 transfectants. We screened 614 sequence-verified clones by Northern blotting, of which 42 genes were scored as over expressed and 17 genes as under expressed in GADD153 transfectants compared with control vector transfectants. Of those genes, 49 corresponded to known genes in public databases. Among them, we further verified that the expression of transferrin (Tf), which is a negative acute-phase protein and is essential to cell survival as a growth factor, was highly modulated by drug-induced GADD153 over expression or by in vitro transfection. GADD153 significantly antagonized the C/EBP (C/EBP-alpha, -beta, and -delta)-mediated transcriptional activation of the Tf gene. In conclusion, Tf and other target genes identified may play a functional role in the downstream pathway of GADD153.

Flixweed (Descurainia sophia L.) is a troublesome weed in winter wheat fields in China. Two flixweed accessions, HB08 and HB16 with a Pro-197-Leu and Pro-197-Ser AHAS-mutation respectively, have evolved very high levels resistance to sulfonylurea (SU) herbicide, tribenuron-methyl. Cross resistance of HB08 and HB16 to AHAS herbicides of SU, imidazolinone (IMI), triazolopyrimidine (TP) and pyrimidinyl-thiobenozoate (PTB) families was investigated by dose-response experiments. In addition, the effects of AHAS herbicides on the activity of AHAS extracted from HB08 and HB16 plants were evaluated. HB16 exhibited cross resistance to SU herbicides halosulfuron-methyl and triasulfuron, TP herbicides flumetsulam and penoxsulam, but displayed more sensitivity to IMI herbicide imazethapyr. By contrast, HB08 only showed cross resistance to SU herbicides halosulfuron-methyl and triasulfuron. The in vitro sensitivity of AHAS to AHAS herbicides is consistent with the results of dose-response experiments and the estimated Pearson’s r values for HB08 and HB16 are 0.996 and 0.912 respectively. These indicated that altered AHAS sensitivity was responsible mainly for cross resistance patterns observed in the two resistant biotypes.

Cadmium (Cd), a toxic heavy metal and carcinogen that is abundantly present in cigarette smoke, is a cause of smoking-induced lung disease. SLC39A8 (ZIP8), a zinc transporter, is a major portal for Cd uptake into cells. We have recently identified that ZIP8 expression is under the transcriptional control of the NF-κB pathway. On the basis of this, we hypothesized that cigarette-smoke induced inflammation would increase ZIP8 expression in lung epithelia, thereby enhancing Cd uptake and cell toxicity. Herein we report that ZIP8 is a central mediator of Cd-mediated toxicity. TNF-α treatment of primary human lung epithelia and A549 cells induced ZIP8 expression, resulting in significantly higher cell death attributable to both apoptosis and necrosis following Cd exposure. Inhibition of the NF-κB pathway and ZIP8 expression significantly reduced cell toxicity. Zinc (Zn), a known cytoprotectant, prevented Cd-mediated cell toxicity via ZIP8 uptake. Consistent with cell culture findings, a significant increase in ZIP8 mRNA and protein expression was observed in the lung of chronic smokers compared with nonsmokers. From these studies, we conclude that ZIP8 expression is induced in lung epithelia in an NF-κB-dependent manner, thereby resulting in increased cell death in the presence of Cd. From this we contend that ZIP8 plays a critical role at the interface between micronutrient (Zn) metabolism and toxic metal exposure (Cd) in the lung microenvironment following cigarette smoke exposure. Furthermore, dietary Zn intake, or a lack thereof, may be a contributing factor in smoking-induced lung disease.