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Figure S5. Primary neurons were either untreated, treated with monomeric α-syn or α-syn fibrils. Seven days later cells were lysed and immunoblots were performed for total levels of syntaxin 1, VAMP2, SNAP25, Synapsin 1, or phospho-Synapsin 1 (site 4/5). The quantitation on the right shows levels of each protein normalized to loading control (vinculin for synapsin 1 and Synapsin 1 for other proteins). The control and fibril exposed neurons represent 6 independent experiments and the monomer exposed neurons represent 3 experiments. There were no significant differences by independent t-test. (PDF 134 kb)

Figure S6. Primary hippocampal control neurons and neurons with inclusions (7 days post-fibril exposure) were incubated with biotin, lysed, and cell surface proteins were pulled down with neutravidin beads. The left immunoblots show cell surface NR2A, NR2B NMDA receptor subunits and GluR1 and GluR2 receptor subunits. The immunoblots on the right show total levels of each protein. Vinculin was included to demonstrate equal loading. Quantitation on the right from 6 independent experiments show the mean level of surface receptor subunits normalized to total levels of each protein. There were no significant differences by independent t-test. (PDF 129 kb)

Tumor-targeted delivery is considered a crucial component of current anticancer drug development and is the best approach to increase the efficacy and reduce the toxicity. Nanomedicine, particularly ligand-based nanoparticles have shown a great potential for active targeting of tumor. Cell penetrating peptide is one of the promising ligands in a targeted cancer therapy. In this study, the gambogic acid-loaded nanostructured lipid carrier (GA-NLC) was modified with two kinds of cell penetrating peptides (cRGD and RGERPPR). The GA-NLC was prepared by emulsification and solvent evaporation method and coupled with cRGD, RGERPPR, and combination cRGD and RGERPPR to form GA-NLC-cRGD, GA-NLC-RGE, and GA-NLC-cRGD/RGE, respectively. The formulations were characterized by their particle size and morphology, zeta potential, encapsulation efficiency, and differential scanning calorimetry. In vitro cytotoxicity and cellular uptake study of the formulations were performed against breast cancer cell (MDA-MB-231). Furthermore, in vivo biodistribution and antitumor activity of the formulations were determined by in vivo imaging and in tumor-bearing nude mice, respectively. The result of in vitro cytotoxicity study showed that GA-NLC-RGE exhibited a significantly higher cytotoxicity on MDA-MB-231 as compared with GA-NLC and GA-Sol. Similarly, RGE-Cou-6-NLC showed remarkably higher uptake by the cells than other NLCs over the incubation period. The in vivo imaging study has demonstrated that among the formulations, the RGE-decorated DiR-NLC were more accumulated in the tumor site. The in vivo antitumor activity revealed that RGE-GA-NLC inhibits the tumor growth more efficiently than other formulations. In conclusion, RGERPPR has a potential as an effective carrier in targeting drug delivery of anticancer agents.

Alterations in insulin granule exocytosis and endocytosis are paramount to pancreatic β cell dysfunction in diabetes mellitus. Here, using temporally controlled gene ablation specifically in β cells in mice, we identified an essential role of dynamin 2 GTPase in preserving normal biphasic insulin secretion and blood glucose homeostasis. Dynamin 2 deletion in β cells caused glucose intolerance and substantial reduction of the second phase of glucose-stimulated insulin secretion (GSIS); however, mutant β cells still maintained abundant insulin granules, with no signs of cell surface expansion. Compared with control β cells, real-time capacitance measurements demonstrated that exocytosis-endocytosis coupling was less efficient but not abolished; clathrin-mediated endocytosis (CME) was severely impaired at the step of membrane fission, which resulted in accumulation of clathrin-coated endocytic intermediates on the plasma membrane. Moreover, dynamin 2 ablation in β cells led to striking reorganization and enhancement of actin filaments, and insulin granule recruitment and mobilization were impaired at the later stage of GSIS. Together, our results demonstrate that dynamin 2 regulates insulin secretory capacity and dynamics in vivo through a mechanism depending on CME and F-actin remodeling. Moreover, this study indicates a potential pathophysiological link between endocytosis and diabetes mellitus.

We evaluated the isoline distribution of thermocline characteristics (the upper and lower boundary temperatures and depths) in the bigeye tuna (Thunnus obesus) fishing grounds in the tropical Atlantic Ocean. We plotted the thermocline characteristics contour on a spatial overlay map using data collected on a monthly basis from Argo buoys and monthly CPUE (catch per unit effort) from bigeye tuna long-lines from the International Commission for the Conservation of Atlantic Tunas (ICCAT). In addition, frequency analysis and the empirical cumulative distribution function (ECDF) were used to calculate the optimum ranges of thermocline characteristics of the central fishing grounds. Our analysis suggested that there were significant seasonal variations in the upper boundary temperature and depth of thermocline in the central fishing grounds, which significantly influenced the temporal and spatial distribution of the bigeye tuna population. The overlay maps suggest that the central fishing ground of the bigeye tuna is located in the enclosed curve region between the higher value (70 m) and the lower value (40 m) of the upper boundary depth, from December to March of each year. After that, the fishing grounds spread to the north and south on both sides. CPUE is highest in areas where the upper boundary depths of the thermocline are between 40 and 60 m to the north of the equator. This extends to between 60 and 100 m to the south of the equator. The CPUE is highest in areas where the upper boundary temperature of the thermocline is between 24 and 26.9 °C in the low-latitude Atlantic Ocean, but decreases to 18 °C offshore of Namibia during July to October. The overlay maps showed that the lower boundary temperature and depth of thermocline have little seasonal variation. The lower boundary depth of the thermocline isoline depth showed that the higher depth area (>250 m) was shaped like the letter ‘W’. Similarly, the lower boundary temperature of thermocline isoline depth suggests that the temperature is higher besides the equator and lower around the equator area. The lower boundary water depth in the central fishing ground of the bigeye tuna ranged between 150 m and 250 m in the low-latitude Atlantic Ocean over the year, whereas the CPUE was low when the depth exceeded 250 m. In addition, the lower boundary water depth in the central fishing ground of the bigeye tuna decreased to between 140 and 160 m offshore of Namibia from April to October. The lower boundary temperature in the central fishing ground was about 12 °C in the low-latitude Atlantic Ocean and 14 °C offshore of Namibia. Conversely, if the temperature is >15 °C, the CPUE tends to be low. The numerical computation results suggest that the optimum ranges for the upper boundary thermocline temperature and depth were between 23 and 26.9 °C, and 30 and 70 m, respectively. The optimum ranges were 12–14.9 °C and 150–230 m for the lower boundary thermocline temperature and depth, respectively. We documented the distribution interval range of thermocline characteristics for the central fishing ground of the bigeye tuna in the Atlantic Ocean. Our results provide a reference for improving the efficiency of the bigeye tuna long-line fleet and aids tuna resource management in the Atlantic Ocean.

Multiple reassortment events within poultry and wild birds had resulted in the establishment of another novel avian influenza A(H10N8) virus, and finally resulted in human death in Nanchang, China. However, there was a paucity of information on the prevalence of avian influenza virus in poultry and wild birds in Nanchang area. We investigated avian influenza virus in poultry and wild birds from live poultry markets, poultry countyards, delivery vehicles, and wild-bird habitats in Nanchang. We analyzed 1036 samples from wild birds and domestic poultry collected from December 2013 to February 2014. Original biological samples were tested for the presence of avian influenza virus using specific primer and probe sets of H5, H7, H9, H10 and N8 subtypes by real-time RT-PCR. In our analysis, the majority (97.98%) of positive samples were from live poultry markets. Among the poultry samples from chickens and ducks, AIV prevalence was 26.05 and 30.81%, respectively. Mixed infection of different HA subtypes was very common. Additionally, H10 subtypes coexistence with N8 was the most prevalent agent during the emergence of H10N8. This event illustrated a long-term surveillance was so helpful for pandemic preparedness and response.

The study population consisted of 105 patients with stage IA to IIB cervical adenocarcinoma (AC) who underwent radical hysterectomy and pelvic lymphadenectomy from three institutions between 1994 and 2015, including 86 patients with bilateral salpingo-oophorectomy (BSO) and 19 patients with ovarian preservation operation. Ovarian metastasis were diagnosed in 3 of 86 patients in BSO group with an incidence rate of 3.5% (3/86). Among the 19 patients with ovarian preservation, none developed an ovarian recurrence in the follow-up (2-71 months). The 5-year overall survival rate of the BSO group and ovarian preservation group were 88.6% and 100%, respectively, with no significant difference (p = .266 > .05). FIGO stage was an independent risk factor of ovarian metastasis for cervical AC (p = .000

Synaptojanin 1 (SJ1) is a major presynaptic phosphatase that couples synaptic vesicle endocytosis to the dephosphorylation of PI(4,5)P2, a reaction needed for the shedding of endocytic factors from their membranes. While the role of SJ1's 5-phosphatase module in this process is well recognized, the contribution of its Sac phosphatase domain, whose preferred substrate is PI4P, remains unclear. Recently a homozygous mutation in its Sac domain was identified in early-onset parkinsonism patients. We show that mice carrying this mutation developed neurological manifestations similar to those of human patients. Synapses of these mice displayed endocytic defects and a striking accumulation of clathrin-coated intermediates, strongly implicating Sac domain's activity in endocytic protein dynamics. Mutant brains had elevated auxilin (PARK19) and parkin (PARK2) levels. Moreover, dystrophic axonal terminal changes were selectively observed in dopaminergic axons in the dorsal striatum. These results strengthen evidence for a link between synaptic endocytic dysfunction and Parkinson's disease.

First of all, from the aspect of key component in the Windows kernel, using the related tools of operating system kernel, to analyze and debug each kernel component by combining with the verification procedures, and objects of individual component in executable level is analyzed deeply, to be familiar with the internal principles of each executable component, and learn to use the kernel debugger, laying the foundations for subsequent in-depth kernel development. Then, this paper studied the techniques commonly used by malicious programs, including the hidden process, images, files and various hook techniques. On this basis, for all kinds of malicious behavior, this paper gives the principle of counter-measures, which is taken by file system filter driven. A file system filter driver module is designed and implemented in this paper. This module realizes the basic encryption and decryption, however a simple XOR operation is used in encryption operation. Because it does not affect research ideas through developing file system filter driver to study the Windows kernel. In the implement of transparent encryption and decryption modules, mainly introduce how to achieve each core routine problem according to the custom data structure combining with the kernel file operation process. The detailed logic flow diagrams and text description are given for each core processing routine. This paper explains basic data structure which is developed by the Windows kernel driver, combing this with the knowledge of the Windows kernel components and the understanding of functional needs permits the customization of a number of important data types. These customized data types include description disk file encryption identification, as well as the process control block in memory that is used to safeguard legitimate processes. The core of this paper is to sort out the processing of files operating the in the kernel, and using this to achieve a core based processing flow of transparent encryption and decryption of code modules.

This paper proposes a new optimized method for fault propagation analysis (FPA) of mechatronic systems. In order to analyze the fault propagation to locate the most vulnerable point, the relevant components of this type of system, such as the software, are abstracted into corresponding models. Firstly, we use a novel graphical representation of a module-signal diagram (MSD) transformed to a signal fault propagation tree (SFPT) and a module fault propagation tree (MFPT). Then fault propagation characteristic parameters are defined to calculate the index of vulnerability for the modules. Meanwhile, a method is presented to find out the most vulnerable path. Finally, a case study of a four-rotor unmanned helicopter is provided to demonstrate and validate the proposed method. By injecting fault to attitude adjustment software modules, we can obtain the fault propagation probability of all modules and the data transfer probability from a module to signals. Then the corresponding SFPT and MFPT can be built from the MSD of the attitude adjustment software system. The final result is the most vulnerable module and path of the example system after performing the proposed method of fault propagation analysis. The result can be used for predicting to locate the fault of mechatronic software systems.

Close appositions between the membrane of the endoplasmic reticulum (ER) and other intracellular membranes have important functions in cell physiology. These include lipid homeostasis, regulation of Ca2+ dynamics, and control of organelle biogenesis and dynamics. Although these membrane contacts have previously been observed in neurons, their distribution and abundance have not been systematically analyzed. Here, we have used focused ion beam-scanning electron microscopy to generate 3D reconstructions of intracellular organelles and their membrane appositions involving the ER (distance ≤30 nm) in different neuronal compartments. ER–plasma membrane (PM) contacts were particularly abundant in cell bodies, with large, flat ER cisternae apposed to the PM, sometimes with a notably narrow lumen (thin ER). Smaller ER–PM contacts occurred throughout dendrites, axons, and in axon terminals. ER contacts with mitochondria were abundant in all compartments, with the ER often forming a network that embraced mitochondria. Small focal contacts were also observed with tubulovesicular structures, likely to be endosomes, and with sparse multivesicular bodies and lysosomes found in our reconstructions. Our study provides an anatomical reference for interpreting information about interorganelle communication in neurons emerging from functional and biochemical studies.

Properties of long persistent luminescence (LPL) and optically stimulated luminescence (OSL) of CaAl2O4:Eu2+, R3+ (R=Nd, Dy, Tm) materials were investigated. The observed phenomenon indicates that R3+ ions (R=Nd, Dy, Tm) have different effects on trap properties of CaAl2O4:Eu2+. The greatly improved LPL performance was observed in Nd3+ co-doped samples, which indicates that the incorporation of Nd3+ creates suitable traps for LPL. While co-doping Tm3+ ions, the intensity of high temperature of thermoluminescence band in CaAl2O4:Eu2+ phosphors is enhanced for the formation of the most suitable traps which benefits the intense and stable OSL. These results suggest that the effective traps contributed to the LPL/OSL are complex, of which could be an aggregation formation with shallow and deep traps other than simple traps from co-doped R3+ ions. The mechanism presented in the end potentially provides explanations of why the OSL of CaAl2O4:Eu2+, R3+ exhibits different read-in/read-out performance as well.

Animal models are critical for gaining insights into autism spectrum disorder (ASD). Despite their apparent advantages to mice for neural studies, rats have not been widely used for disorders of the human CNS, such as ASD, for the lack of convenient genome manipulation tools. Here we describe two of the first transgenic rat models for ASD, developed using zinc-finger nuclease (ZFN) methodologies, and their initial behavioral assessment using a rapid juvenile test battery. A syndromic and nonsyndromic rat model for ASD were created as two separate knockout rat lines with heritable disruptions in the genes encoding Fragile X mental retardation protein (FMRP) and Neuroligin3 (NLGN3). FMRP, a protein with numerous proposed functions including regulation of mRNA and synaptic protein synthesis, and NLGN3, a member of the neuroligin synaptic cell-adhesion protein family, have been implicated in human ASD. Juvenile subjects from both knockout rat lines exhibited abnormalities in ASD-relevant phenotypes including juvenile play, perseverative behaviors, and sensorimotor gating. These data provide important first evidence regarding the utility of rats as genetic models for investigating ASD-relevant genes.

Axonal lysosomes accumulate abnormally in Alzheimer’s disease brains. However, whether and how such lysosomes contribute to disease pathology has been unclear. Gowrishankar et al. show that the JIP3-dependent transport of axonal lysosomes negatively regulates amyloid precursor protein processing into amyloidogenic peptides., Lysosomes robustly accumulate within axonal swellings at Alzheimer’s disease (AD) amyloid plaques. However, the underlying mechanisms and disease relevance of such lysosome accumulations are not well understood. Motivated by these problems, we identified JNK-interacting protein 3 (JIP3) as an important regulator of axonal lysosome transport and maturation. JIP3 knockout mouse neuron primary cultures accumulate lysosomes within focal axonal swellings that resemble the dystrophic axons at amyloid plaques. These swellings contain high levels of amyloid precursor protein processing enzymes (BACE1 and presenilin 2) and are accompanied by elevated Aβ peptide levels. The in vivo importance of the JIP3-dependent regulation of axonal lysosomes was revealed by the worsening of the amyloid plaque pathology arising from JIP3 haploinsufficiency in a mouse model of AD. These results establish the critical role of JIP3-dependent axonal lysosome transport in regulating amyloidogenic amyloid precursor protein processing and support a model wherein Aβ production is amplified by plaque-induced axonal lysosome transport defects.

Lowe syndrome and Dent disease are two conditions that result from mutations of the inositol 5-phosphatase oculocerebrorenal syndrome of Lowe (OCRL) and share the feature of impaired kidney proximal tubule function. Genetic ablation of Ocrl in mice failed to recapitulate the human phenotypes, possibly because of the redundant functions of OCRL and its paralog type 2 inositol polyphosphate-5-phosphatase (INPP5B). Germline knockout of both paralogs in mice results in early embryonic lethality. We report that kidney tubule-specific inactivation of Inpp5b on a global Ocrl-knockout mouse background resulted in low molecular weight proteinuria, phosphaturia, and acidemia. At the cellular level, we observed a striking impairment of clathrin-dependent and -independent endocytosis in proximal tubules, phenocopying what has been reported for Dent disease caused by mutations in the gene encoding endosomal proton-chloride exchange transporter 5. These results suggest that the functions of OCRL/INPP5B and proton-chloride exchange transporter 5 converge on shared mechanisms, the impairment of which has a dramatic effect on proximal tubule endocytosis.

From the concept of software dynamic evolution, the paper makes a detail analysis of the influence of the software evolution on software mission reliability. The basic question of the software mission reliability for the software evolution is presented. The model framework of software mission reliability is built, and the modeling methods of software mission reliability models are given. The research conclusions have importantly theoretical and engineering value for the reliability analysis and evaluation of the reconfigurable software on function.

Zr60-xAl15Co25Tix (x = 0, 5, 10, and 15 at%) coatings were fabricated on Q235 steel by laser cladding. The influence of Ti substitution on the properties of the coated specimens was evaluated in terms of microstructure, hardness, and corrosion resistance. Results show that the coating exhibited a sound metallurgical bonding to the substrate. The cladding layer composed of amorphous and nanocrystalline phases led to increased hardness and enhanced corrosion resistance compared with the substrate. Microhardness values of the coatings increased with the substitution of Ti for Zr attributing to the higher content of ZrO2 while lower content of amorphous phase. Both the corrosion potential and corrosion current density show that appropriate addition of Ti (5 at%) can further improve the corrosion resistance. The results offer a better understanding of the development and application of Zr-based amorphous coating for corrosive environment.

In the present work, mono-disperse and uniform orthorhombic lutetium fluoride (LuF3) nanocrystals with an average size of about 35 nm have been successfully synthesized by a simple ionothermal method without any template. The infrared (IR) to visible up-conversion (UC) photoluminescence of LuF3 doped with Yb3+, Tm3+, and Ho3+ under 980 nm excitation was systemically studied. The intensity of near infrared (NIR) to visible up-conversion emission of Tm3+ was improved efficiently by adding Yb3+ and Ho3+ in LuF3, especially for the broad NIR emission band located at 812 nm. Meanwhile, compared to the Yb3+ and Tm3+ co-doped LuF3, the ratio of red to green emission in the Yb3+, Tm3+, and Ho3+ co-doped LuF3 changed greatly, and a bright yellowish-green emission was observed under 980 nm laser excitation. It shows that Yb3+, Tm3+ and Ho3+ co-doped LuF3 nanocrystals provided a potential application in vitro and in vivo bio-imaging, color displays and optical storage.

The analysis of high resolution (6m) radar data was carried out on the Antarctic sea ice segmentation. This paper presented two methods for Synthetic Aperture Radar (SAR) sea ice image segmentation: k-means clustering method and threshold value method (binary). First, the speckle noise in SAR image was filtered and segmented by two methods. Then the results were compared. It was found that both methods could distinguish the crack and small ice, and these two sea ice distribution results were almost similar. The difference between two methods was: image of threshold value method contained disordered spots and isolated small ice, while k-mean method outcome had clear segmentation boundary and ensured completely segmentation region. So the result from the k-mean clustering method was better. In order to reflect the advantage of high resolution image, the image processed by k-mean clustering method was contrast with Antarctic sea ice concentration data, which was received from Advanced Microwave Scanning Radiometer 2. It is fully proved that the high resolution radar data can provide the location and the size of sea ice distribution more clearly and more accurately.

SO2 capacity of the obtained TMG-AlPO-5/cordierite honeycomb ceramic (CHC) adsorbent was measured to be 1.13 mol per mol TMG. More importantly, compared with literature reported supported ionic liquids, it is featured by a significantly improved adsorption rate (t0.9 reduced from >30 min to ∼0.1 min) and negligible pressure drop.

The rat is a preferred model system over the mouse for neurological studies, and cell type-specific Cre expression in the rat enables precise ablation of gene function in neurons of interest, which is especially valuable for neurodegenerative disease modeling and optogenetics. Yet, few such Cre rats are available. Here we report the characterization of two Cre rats, tyrosine hydroxylase (TH)-Cre and dopamine active transporter (DAT or Slc6a3)-Cre, by using a combination of immunohistochemistry (IHC) and mRNA fluorescence in situ hybridization (FISH) as well as a fluorescent reporter for Cre activity. We detected Cre expression in expected neurons in both Cre lines. Interestingly, we also found that in Th-Cre rats, but not DAT-Cre rats, Cre is expressed in female germ cells, allowing germline excision of the floxed allele and hence the generation of whole-body knockout rats. In summary, our data demonstrate that targeted integration of Cre cassette lead to faithful recapitulation of expression pattern of the endogenous promoter, and mRNA FISH, in addition to IHC, is an effective method for the analysis of the spatiotemporal gene expression patterns in the rat brain, alleviating the dependence on high quality antibodies that are often not available against rat proteins. The Th-Cre and the DAT-Cre rat lines express Cre in selective subsets of dopaminergic neurons and should be particularly useful for researches on Parkinson's disease.

Recent studies reported that long non-coding RNAs (lncRNAs) might play critical roles in regulating endocrine resistance of breast cancer. Urothelial carcinoma-associated 1 (UCA1) is an lncRNA with an oncogenic role in breast cancer. This study aimed to investigate whether UCA1 is involved in acquired tamoxifen resistance in estrogen receptor (ER)-positive cancer cells. Our findings reveal that tamoxifen induces UCA1 upregulation in ER-positive breast cancer cells in a HIF1α-dependent manner. UCA1 upregulation results in significantly enhanced tamoxifen resistance. The upregulated UCA1 sponges miR-18a, which is a negative regulator of HIF1α. Therefore, UCA1 upregulation is further enhanced through a miR-18a-HIF1α feedback loop. In addition, our data also showed that miR-18a is a modulator of tamoxifen sensitivity due to its regulative effect on cell cycle proteins. miR-18a inhibitor reduced the sensitivity of MCF-7 cells to tamoxifen, while miR-18a mimics sensitized BT474 cells to tamoxifen. Therefore, miR-18a downregulation also partly contributes to acquired tamoxifen resistance in the cancer cells. These findings provide some useful information for future clinical treatment of tamoxifen resistance.

Chinese oceanic fisheries have already covered seven main ocean area, including East Pacific, West Pacific, Middle Atlantic, North Pacific, Southeast Pacific, Southwest Atlantic and Indian Ocean. Based on the ocean environment data obtained from remote sensing and fishing historical data, the fishing ground analysis and forecasting information system has been constructed for Chinese oceanic fisheries. This paper briefly described the concerned remote sensing data, system architecture, client and models. Then a case application for tuna of the Indian Ocean was given at the end of the paper. The whole system has already in operation as making and delivering the forecasting information product every week.

Two typical satellite sea surface temperature (SST) datasets, from the Multi-functional Transport Satellite (MTSAT) and Tropical Rainfall Measuring Mission Microwave Imager (TMI), were evaluated for the East China Sea, Yellow Sea, and Bohai Sea throughout 2008. Most monthly-mean availabilities of MTSAT are higher than those of TMI, whereas the seasonal variation of the latter is less than that of the former. The analysis on the one-year data shows that the annual mean availability of MTSAT (61%) is greater than that of TMI (56%). This is mainly because MTSAT is a geostationary satellite, which achieves longer observation than the sun-synchronous TMI. The daily availability of TMI (28%–75%) is more constant than that of MTSAT (9%–93%). The signal of infrared sensors on MTSAT is easily disturbed on cloudy days. In contrast, the TMI microwave sensor can obtain information through clouds. Based on in-situ SSTs, the SST accuracy of TMI is superior to that of MTSAT. In 2008, the root mean square (RMS) error of TMI and MTSAT were 0.77 K and 0.84 K, respectively. The annual mean biases were 0.14 K (TMI) and −0.31 K (MTSAT). To attain a high availability of SSTs, we propose a fusion method to merge both SSTs. The annual mean availability of fusion SSTs increases 17% compared to MTSAT. In addition, the availabilities of the fusion SSTs become more constant. The annual mean RMS and bias of fusion SSTs (0.78 K and −0.06 K, respectively) are better than those of MTSAT (0.84 K and −0.31 K).

A common deletion comprising LCE3B and LCE3C, members of the late cornified envelope (LCE) gene cluster, has been shown to be significantly associated with psoriasis in several Caucasian populations. The expression of LCE can be induced by skin barrier disruption, leading to psoriatic lesions. To identify whether deletion of genes in the LCE region is a genetic risk factor in the pathogenesis of psoriasis, we genotyped the LCE3C and LCE3B deletion and single-nucleotide polymorphism rs4112788, which is in strong linkage disequilibrium with the LCE gene cluster, via direct sequencing in 468 psoriasis patients and 768 controls in a Chinese population. We found that deletion of the two LCE genes was associated with psoriasis (odds ratio=1.917; 95% confidence interval=1.291-2.847, P=0.001), a conclusion that was similar to that of another independent Chinese cohort study. The deletion was not significantly associated with the age of disease onset, and there was no significant epistatic interaction between deletion and PSORS1 risk allele on 6p21.3. Our study confirms an association between the deletion of LCE3C and LCE3B and psoriasis in a Chinese population.

A sequence of 11 consecutive arginine residues (11R) is one of the best protein transduction domains for introducing proteins into cell membranes. Heme-oxygenase-1 (HO-1) is involved in heme catabolism and reduces the contractile effect of hemoglobin after subarachnoid hemorrhage (SAH). Therefore, we constructed 11R-fused HO-1 protein to achieve successful transduction of the protein into the cerebral arteries and examined the therapeutic effect of the 11R-HO-1 protein for cerebral vasospasm (CV) after SAH. We injected the 11R-HO-1 protein into the cisterna magna of male rats and, several hours after the injection, performed immunofluorescence staining and western blotting analysis of the rat basilar arteries (BAs) to determine transduction efficacy. We also assessed intraarterial HO-1 activity as cGMP (cyclic guanosine 3′, 5′-cyclic monophosphate) accumulation in SAH and determined whether protein transduction of 11R-HO-1 quantified the therapeutic effect in a rat double-hemorrhage model of SAH. The BAs expressed significantly more HO-1 in the group injected with 11R-HO-1 (3.56±0.54 (11R-HO-1) versus control (saline)), and transduction of 11R-HO-1 resulted in higher activity (>3.25-fold) in rat BAs with SAH. Moreover, the results of the rat double-hemorrhage model showed that the 11R-HO-1 protein significantly attenuated CV after SAH (317.59±23.48 μm (11R-HO-1) versus 270.08±14.66 μm (11R-fused enhanced green fluorescent protein), 252.05±13.95 μm (saline), P

The sea surface temperature (SST) of the East China Sea (ECS) increased in the past decades, which may have a great impact on the ecosystem of the ECS, including the changes in planktonpopulation structure. In this paper, the changes in peaked abundance of Calanus sinicus in the Changjiang River (Yangtze River) Estuary were compared between 1959 and 2002, based on the data collected from the seasonally oceanographic cruises and those performed in spring of 2005. It was much higher in spring compared with that in other seasons both in 1959 and 2002. Furthermore, in spring 2005, the time for occurrence and decrease of the peaked C. sinicus abundance advanced about one month, accompanying the increase in the sea surface water temperature (SST). It peaked in June and decreased in July in 1959, however, in 2005, it peaked in May and attenuated sharply in early June. The earlier decrease of peaked C. sinicus abundance may further deteriorate the ecosystem in the Changjiang River Estuary and north nearshore of the ECS.

Lymphocyte homing is regulated by the dynamic interaction between integrins and their ligands. Integrin α4β7 mediates both rolling and firm adhesion of lymphocytes by modulating its affinity to the ligand, mucosal addressin cell adhesion molecule-1 (MAdCAM-1). Although previous studies have revealed some mechanisms of α4β7-MAdCAM-1 binding, little is known about the different molecular bases of the low- and high-affinity α4β7-MAdCAM-1 interactions, which mediate rolling and firm adhesion of lymphocytes, respectively. Here, we found that two loops in immunoglobulin domains 1 and 2 (D1 and D2) of MAdCAM-1 played different roles in MAdCAM-1 binding to low-affinity (inactive) and high-affinity (activated) α4β7. The Asp-42 in the CC′ loop of D1 was indispensable for MAdCAM-1 binding to both low-affinity and high-affinity α4β7. The other CC′ loop residues except for Arg-39 and Ser-44 were essential for MAdCAM-1 binding to both inactive α4β7 and α4β7 activated by SDF-1α or talin, but not required for MAdCAM-1 binding to Mn2+-activated α4β7. Single amino acid substitution of the DE loop residues mildly decreased MAdCAM-1 binding to both inactive and activated α4β7. Notably, removal of the DE loop greatly impaired MAdCAM-1 binding to inactive and SDF-1α- or talin-activated α4β7, but only decreased 60% of MAdCAM-1 binding to Mn2+-activated α4β7. Moreover, DE loop residues were important for stabilizing the low-affinity α4β7-MAdCAM-1 interaction. Thus, our findings demonstrate the distinct roles of the CC′ and DE loops in the recognition of MAdCAM-1 by low- and high-affinity α4β7 and suggest that the inactive α4β7 and α4β7 activated by different stimuli have distinct conformations with different structural requirements for MAdCAM-1 binding.

Gene targeting is indispensible for reverse genetics and the generation of animal models of disease. The mouse has become the most commonly used animal model system owing to the success of embryonic stem cell-based targeting technology, whereas other mammalian species lack convenient tools for genome modification. Recently, microinjection of engineered zinc-finger nucleases (ZFNs) in embryos was used to generate gene knockouts in the rat and the mouse by introducing nonhomologous end joining (NHEJ)-mediated deletions or insertions at the target site. Here we use ZFN technology in embryos to introduce sequence-specific modifications (knock-ins) by means of homologous recombination in Sprague Dawley and Long-Evans hooded rats and FVB mice. This approach enables precise genome engineering to generate modifications such as point mutations, accurate insertions and deletions, and conditional knockouts and knock-ins. The same strategy can potentially be applied to many other species for which genetic engineering tools are needed.

Psoriasis is an inflammatory skin disease with a multifactorial genetic basis. A recent genome-wide association study identified several psoriasis-predisposing loci, including IL-12B, which encodes the common p40 subunit of interleukin (IL)-12 and IL-23 and the IL-23 receptor gene (IL-23R). To investigate the relationships of these predisposing polymorphisms with psoriasis in the Chinese Han population, we genotyped three representative variants (rs6887695, rs11465817, and rs1343152) in 217 unrelated patients and 288 control subjects using direct sequencing, We further replicated the positive polymorphism, rs6887695, in a larger combined sample that included 578 patients and 1422 controls. We found the single nucleotide polymorphism, rs6887695 (odds ratio = 1.33, 95% confidence interval 1.03-1.73, p = 0.028) of IL-2B to be significantly associated with psoriasis, and a novel halotype A-A of rs11465817-rs1343152 also showed a positive association. Our study confirms the effects of IL-12B and IL-23R variants on psoriasis in East Asian populations, and provides a reference point for further investigation of the role of the IL-12/IL-23 pathway in chronic epithelial inflammation in Asian and other ethnic populations.

To obtain information on the outer surface of a cylinder object, we propose a catadioptric panoramic imaging system based on the principle of uniform spatial resolution for vertical scenes. First, the influence of the projection-equation coefficients on the spatial resolution and astigmatism of the panoramic system are discussed, respectively. Through parameter optimization, we obtain the appropriate coefficients for the projection equation, and so the imaging quality of the entire imaging system can reach an optimum value. Finally, the system projection equation is calibrated, and an undistorted rectangular panoramic image is obtained using the cylindrical-surface projection expansion method. The proposed 360-deg panoramic-imaging device overcomes the shortcomings of existing surface panoramic-imaging methods, and it has the advantages of low cost, simple structure, high imaging quality, and small distortion, etc. The experimental results show the effectiveness of the proposed method.

In this paper, we apply Anti-de Sitter (AdS) black hole solution of the Einstein-æther theory to the study of the holographic superconductor and show that the AdS black hole solution can be rewritten in some very simple forms, from which it is easy to identify the locations of various killing horizons. Then, we investigate the different effects of these horizons on the holographic superconductor.

The relationships between the neon flying squid, Ommastrephes bartrami, and the relative ocean environmental factors are analyzed. The environmental factors collected are sea surface temperature (SST), chlorophyll concentration (Chl-a) and sea surface height (SSH) from NASA, as well as the yields of neon flying squid in the North Pacific Ocean. The results show that the favorable temperature for neon flying squid living is 10°C–22°C and the favorite temperature is between 15°C–17°C. The Chl-a concentration is 0.1–0.6 mg/m3. When Chl-a concentration changes to 0.12–0.14 mg/m3, the probability of forming fishing ground becomes very high. In most fishing grounds, the SSH is higher than the mean SSH. The generalized additive model (GAM) was applied to analyze the correlations between neon flying squid and ocean environmental factors. Every year, squids migrate northward from June to August and return southward during October–November, and the characteristics of the both migrations are very different. When squids migrate to the north, most relationships between the yields and SST are positive. The relationships are negative when squids move to southward. The relationships between the yields and Chl-a concentrations are negative from June to October, and insignificant in November. There is no obvious correlation between the catches of squid and longitude, but good with latitude.

Anisotropic fluid with self-similarity of the second kind in (2 + 1)-dimensional space–times with circular symmetry is studied. By imposing the condition that the radial pressure vanishes, we show that the only allowed solutions are the ones that represent dust fluid. All such solutions to the Einstein field equations are found and their local and global properties are studied in detail. It is found that some can be interpreted as representing gravitational collapse, in which both naked singularities and black holes can be formed.

Previous studies have demonstrated that histamine (HA) acts as a neurotransmitter in the cardiac sympathetic nervous system of the guinea pig. The aim of the current study was to examine whether HA widely exists in the sympathetic nervous systems of other species and the subcellular localization of HA in sympathetic terminals. An immunofluorescence histochemical multiple-staining technique and anterograde tracing method were employed to visualize the colocalization of HA and norepinephrine (NE) in sympathetic ganglion and nerve fibers in different species. Pre-embedding immunoelectron microscopy was used to observe the subcellular distribution of HA in sympathetic nerve terminals. Under the confocal microscope, coexistence of NE and HA was displayed in the superior cervical ganglion and celiac ganglion neurons of the mouse and dog as well as in the vas deferens, mesenteric artery axon, and varicosities of the mouse and guinea pig. Furthermore, colocalization of NE and HA in cardiac sympathetic axons and varicosities was labeled by biotinylated dextranamine injected into the superior cervical ganglion of the guinea pig. By electron microscopy, HA-like high-density immunoreactive products were seen in the small vesicles of the guinea pig vas deferens. These results provide direct cellular and subcellular morphological evidence for the colocalization of HA and NE in sympathetic ganglion and nerve fibers, and support that HA is classified as a neurotransmitter in sympathetic neurons.

Under normal physiological conditions, synaptic vesicle endocytosis is regulated by phosphorylation and Ca(2+)-dependent dephosphorylation of endocytic proteins such as amphiphysin and dynamin. To investigate the regulatory mechanisms that may occur under the conditions of excessive presynaptic Ca(2+) influx observed preceding neural hyperexcitation, we examined hippocampal slices following high-potassium or high-frequency electrical stimulation (HFS). In both cases, three truncated forms of amphiphysin I resulted from cleavage by the protease calpain. In vitro, the binding of truncated amphiphysin I to dynamin I and copolymerization into rings with dynamin I were inhibited, but its interaction with liposomes was not affected. Moreover, overexpression of the truncated form of amphiphysin I inhibited endocytosis of transferrin and synaptic vesicles. Inhibiting calpain prevented HFS-induced depression of presynaptic transmission. Finally, calpain-dependent amphiphysin I cleavage attenuated kainate-induced seizures. These results suggest that calpain-dependent cleavage of amphiphysin I inhibits synaptic vesicle endocytosis during neural hyperexcitation and demonstrate a novel post-translational regulation of endocytosis.

The four highly conserved Notch receptors receive short-range signals that control many biological processes during development and in adult vertebrate tissues. The involvement of Notch1 signaling in tissue self-renewal is less clear, however. We developed a novel genetic approach N1IP-CRE(Notch1 Intramembrane Proteolysis) to follow, at high resolution, the descendents of cells experiencing Notch1 activation in the mouse. By combining N1IP-CRE with loss-of-function analysis, Notch activation patterns were correlated with function during development, self-renewal and malignancy in selected tissues. Identification of many known functions of Notch1 throughout development validated the utility of this approach. Importantly,novel roles for Notch1 signaling were identified in heart, vasculature, retina and in the stem cell compartments of self-renewing epithelia. We find that the probability of Notch1 activation in different tissues does not always indicate a requirement for this receptor and that gradients of Notch1 activation are evident within one organ. These findings highlight an underappreciated layer of complexity of Notch signaling in vivo. Moreover, NIP-CRE represents a general strategy applicable for monitoring proteolysis-dependent signaling in vivo.

A new algorithm based on the multiparameter neural network is proposed to retrieve wind speed (WS), sea surface temperature (SST), sea surface air temperature, and relative humidity (RH) simultaneously over the global oceans from Special Sensor Microwave Imager (SSM/I) observations. The retrieved geophysical parameters are used to estimate the surface latent heat flux and sensible heat flux using a bulk method over the global oceans. The neural network is trained and validated with the matchups of SSM/I overpasses and National Data Buoy Center buoys under both clear and cloudy weather conditions. In addition, the data acquired by the 85.5-GHz channels of SSM/I are used as the input variables of the neural network to improve its performance. The root-mean-square (rms) errors between the estimated WS, SST, sea surface air temperature, and RH from SSM/I observations and the buoy measurements are 1.48 m s−1, 1.54°C, 1.47°C, and 7.85, respectively. The rms errors between the estimated latent and sensible heat fluxes from SSM/I observations and the Xisha Island (in the South China Sea) measurements are 3.21 and 30.54 W m−2, whereas those between the SSM/I estimates and the buoy data are 4.9 and 37.85 W m−2, respectively. Both of these errors (those for WS, SST, and sea surface air temperature, in particular) are smaller than those by previous retrieval algorithms of SSM/I observations over the global oceans. Unlike previous methods, the present algorithm is capable of producing near-real-time estimates of surface latent and sensible heat fluxes for the global oceans from SSM/I data.

To analyse the necessity of colposcopic directed biopsy to vaginal intraepithelial neoplasia (VAIN) before hysterectomy due to early stage cervical cancer (stage I) or high grade cervical intraepithelial neoplasia (CIN).A total of 669 patients who underwent a hysterectomy due to early stage cervical cancer (stage I) and CIN III in Beijing Obstetrics and Gynecology Hospital, Capital Medical University, from January 1, 2009 to December 31, 2013 and followed up, 99 patients with VAIN were enrolled. The clinical data and following up the prognosis were prospectively analyzed retrospectively.The occurrence rate of VAIN before and after hysterectomy due to cervical dysplasia was 14.8% (99/669), the occurrence rate and the grade of VAIN showed that significantly increased from CIN III to cervical cancer stage I (P0.05); Only 15 patients enrolled had undergone vaginal wall biopsy by colposcopy pre-hysterectomy, including 11 patients who were diagnosed with VAIN II-III and underwent vagina extended resection during the hysterectomy. The 5 year recurrence rate of vaginal stump VAIN after hysterectomy was 12.1% (12/99) and the progression rate was 4.0% (4/99), the recurrent rate was 2.0% (2/99).For all the patients who are planning to undergo hysterectomy due to stage I cervical cancer and CIN III, routine upper side of the vagina wall colposcopic-directed biopsy prehysterectomy is strongly recommended. All the patients after hysterectomy due to cervical dysplasia should be followed up regularly within 3 years after hysterectomy.

Through a comprehensive analysis of organellar markers in mouse models of Alzheimer’s disease, we document a massive accumulation of lysosome-like organelles at amyloid plaques and establish that the majority of these organelles reside within swollen axons that contact the amyloid deposits. This close spatial relationship between axonal lysosome accumulation and extracellular amyloid aggregates was observed from the earliest stages of β-amyloid deposition. Notably, we discovered that lysosomes that accumulate in such axons are lacking in multiple soluble luminal proteases and thus are predicted to be unable to efficiently degrade proteinaceous cargos. Of relevance to Alzheimer’s disease, β-secretase (BACE1), the protein that initiates amyloidogenic processing of the amyloid precursor protein and which is a substrate for these proteases, builds up at these sites. Furthermore, through a comparison between the axonal lysosome accumulations at amyloid plaques and neuronal lysosomes of the wild-type brain, we identified a similar, naturally occurring population of lysosome-like organelles in neuronal processes that is also defined by its low luminal protease content. In conjunction with emerging evidence that the lysosomal maturation of endosomes and autophagosomes is coupled to their retrograde transport, our results suggest that extracellular β-amyloid deposits cause a local impairment in the retrograde axonal transport of lysosome precursors, leading to their accumulation and a blockade in their further maturation. This study both advances understanding of Alzheimer’s disease brain pathology and provides new insights into the subcellular organization of neuronal lysosomes that may have broader relevance to other neurodegenerative diseases with a lysosomal component to their pathology.