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Considering that photodynamic therapy (PDT)-induced oxygen consumption and microvascular damage could exacerbate hypoxia to drive more glycolysis and angiogenesis, a novel approach to potentiate PDT and overcome the resistances of hypoxia is avidly needed. Herein, morpholine-modified PEGylated bilirubin was proposed to co-deliver chlorin e6, a photosensitizer, and diclofenac (Dc). In acidic milieu, the presence of morpholine could enable the nanocarriers to selectively accumulate in tumor cells, while PDT-generated reactive oxidative species (ROS) resulted in the collapse of bilirubin nanoparticles and rapid release of Dc. Combining with Dc showed a higher rate of apoptosis over PDT alone and simultaneously triggered a domino effect, including blocking the activity and expression of lactate dehydrogenase A (LDHA), interfering with lactate secretion, suppressing the activation of various angiogenic factors and thus obviating hypoxia-induced resistance-glycolysis and angiogenesis. In addition, inhibition of hypoxia-inducible factor-1

While research on cancer vaccines has made great strides in the field of immunotherapy, the targeted delivery of multiple effective components (rational-tailored antigens and adjuvants) remains a challenge. Here, we utilized the unique hierarchical structures of Pickering emulsions (particles, oil core, and water-oil interface) to develop mannosylated (M) Pickering emulsions (PE) that target antigen presenting cells and synergistically deliver antigenic peptides and the TLR9 agonist CpG (C) as an enhanced cancer vaccine (MPE-C). We chemically linked mannose residues to PLGA/PLAG-PEG nanoparticles and produced a dense array of mannose on the nanopatterned surface of Pickering emulsions, allowing for increased cellular targeting. Together with the inherent deformability of the oily core, MPE-C increased the droplet-cellular contact area and provoked the cellular recognition of mannose and CpG for enhanced immune activation. We found that MPE-C attracted a large number of APCs to the local site of administration, evidently increasing cellular uptake and activation. Additionally, we observed increased antigen-specific cellular immune responses, with potent anti-tumor effects against both E.G7-OVA and B16-MUCI tumors. Furthermore, MPE-C combined with PD-1 antibodies produced a significant tumor regression, resulting in synergistic increases in anti-tumor effects. Thus, through the strategic loading of mannose, antigens, and CpG, Pickering emulsions could serve as a targeted delivery platform for enhanced multicomponent cancer vaccines.

Increasing uric acid (UA) could induce renal tubular epithelial cell (NRK-52E) injury. However, the specific mechanism by which UA induces renal tubular epithelial cell injury remains unknown. It was hypothesized that UA induces renal tubular epithelial cell injury through reactive oxygen species (ROS) and the Never in mitosis gene A (NIMA)-related kinase 7 (NEK7)/NLR family pyrin domain containing 3 (NLRP3) signaling pathway. TUNEL assay and flow cytometry were applied to measure apoptosis, and the results of the present study showed that UA treatment induced apoptosis of NRK-52E cells in a concentration-dependent manner. Western blotting was performed to determine the expression levels of cleaved caspase-3, Bax and Bcl-xl, it was found that levels were significantly increased after UA treatment in NRK-52E cells. ROS and apoptosis were predominantly induced in NRK-52E cells and there was an association between ROS and apoptosis. Enhanced expression of NEK7, NLRP3, apoptosis-associated speck-like and caspase-1 were observed in NRK-52E cells treated with UA. The ROS inhibitor, N-acetyl-l-cysteine, exerted a protective effect on the UA-induced apoptosis of tubular epithelial cells by reducing excess ROS production, which significantly inhibited NEK7 and NLRP3 inflammasome activation. These results indicated that UA activates ROS and induces apoptosis of NRK-52E cells. The mechanism might be related to the regulation of the NEK7/NLRP3 signaling pathway.

MicroRNAs (miRNAs) are involved in lymphoma progression by regulating the tumor microenvironment. Serum miR130b is overexpressed in diffuse large B-cell lymphoma (DLBCL), inducing Th17 cell alterations. To further illustrate its biological significance and therapeutic rationale, miR130b was detected by quantitative real-time PCR in the serum samples of 532 newly diagnosed DLBCL patients. The mechanism of miR130b on lymphoma progression and the tumor microenvironment was investigated both in vitro and in vivo. Therapeutic targeting miR130b was also evaluated, including OX40 agonistic antibody and lipid nanoparticles (LNPs)-miR130b antagomir. The results showed that serum miR130b significantly correlated with tumor miR130b and serum interleukin-17, indicating lymphoma relapse and inferior survival of DLBCL patients. MiR130b overexpression altered tumor microenvironment signaling pathways and increased Th17 cell activity. As mechanism of action, miR130b downregulated tumor OX40L expression by directly targeting IFNAR1/p-STAT1 axis, recruiting Th17 cells via OX40/OX40L interaction, thereby promoting immunosuppressive function of Th17 cells. In co-culture systems of B-lymphoma cells with immune cells, miR130b inhibited lymphoma cell autophagy, which could be counteracted by OX40 agonistic antibody and LNPs-miR130b antagomir. In murine xenograft model established with subcutaneous injection of A20 cells, both OX40 agonistic antibody and LNPs-miR130b antagomir remarkably inhibited Th17 cells and retarded miR130b-overexpressing tumor growth. In conclusion, as an oncogenic biomarker of DLBCL, miR130b was related to lymphoma progression through modulating OX40/OX40L-mediated lymphoma cell interaction with Th17 cells, attributing to B-cell lymphoma sensitivity towards OX40 agonistic antibody. Targeting miR130b using LNPs-miR130b antagomir could also be a potential immunotherapeutic strategy in treating OX40-altered lymphoid malignancies.

Vps13 family proteins are proposed to function in bulk lipid transfer between membranes, but little is known about their regulation. During sporulation of Saccharomyces cerevisiae, Vps13 localizes to the prospore membrane (PSM) via the Spo71–Spo73 adaptor complex. We previously reported that loss of any of these proteins causes PSM extension and subsequent sporulation defects, yet their precise function remains unclear. Here, we performed a genetic screen and identified genes coding for a fragment of phosphatidylinositol (PI) 4-kinase catalytic subunit and PI 4-kinase noncatalytic subunit as multicopy suppressors of spo73Δ. Further genetic and cytological analyses revealed that lowering PI4P levels in the PSM rescues the spo73Δ defects. Furthermore, overexpression of VPS13 and lowering PI4P levels synergistically rescued the defect of a spo71Δ spo73Δ double mutant, suggesting that PI4P might regulate Vps13 function. In addition, we show that an N-terminal fragment of Vps13 has affinity for the endoplasmic reticulum (ER), and ER-plasma membrane (PM) tethers localize along the PSM in a manner dependent on Vps13 and the adaptor complex. These observations suggest that Vps13 and the adaptor complex recruit ER-PM tethers to ER-PSM contact sites. Our analysis revealed that involvement of a phosphoinositide, PI4P, in regulation of Vps13, and also suggest that distinct contact site proteins function cooperatively to promote de novo membrane formation., Author summary Vps13 family proteins are conserved lipid transfer proteins that function at organelle contact sites and have been implicated in a number of different neurological diseases. In the yeast Saccharomyces cerevisiae, Vps13 is encoded by a single gene and is localized to various contact sites by interaction with different adaptor proteins and/or lipids, however its regulation is yet to be clarified. We have previously shown that during the developmental process of sporulation, Vps13 is recruited to de novo membrane structures called prospore membranes (PSMs) by a specific adaptor complex, and Vps13 and its adaptors are required for PSM extension. Here we reveal that loss of an adaptor can be overcome by lowering phosphatidylinositol-4-phosphate (PI4P) levels, either by inhibiting PI 4-kinase on the PSM or recruiting PI 4-phospatase to the PSM and that PI4P levels in the PSM affect Vps13 function. Further, we show that Vps13 forms endoplasmic reticulum (ER)-PSM contact sites, that ER-plasma membrane tethering proteins are recruited to ER-PSM contacts, and these proteins may function in conjunction with Vps13. Thus, our work shines light on both the mechanisms of intracellular remodeling and the function of this important class of lipid transfer proteins.

Small cell lung cancer (SCLC) is a recalcitrant cancer featured with high metastasis. However, the exact cell type contributing to metastasis remains elusive. UsingRb1L/L/Trp53L/Lmouse model, we identify the NCAMhiCD44lo/−subpopulation as SCLC metastasizing cell (SMC), which is progressively transitioned from non-metastasizing NCAMloCD44hicell (Non-SMC). Integrative chromatin accessibility and gene expression profiling studies reveal an important role of SWI/SNF complex, and knockout of its central component,Brg1, significantly inhibits such phenotypic transition and metastasis. Mechanistically, TAZ is silenced by SWI/SNF complex during SCLC malignant progression, and its knockdown promotes SMC transition and metastasis. Importantly, ectopic TAZ expression reversely drives SMC-to-Non-SMC transition and alleviates metastasis. Single-cell RNA-sequencing analyses identify SMC as the dominant subpopulation in human SCLC metastasis, and immunostaining data show a positive correlation between TAZ and patient prognosis. These data uncover high SCLC plasticity and identify TAZ as key molecular switch in orchestrating SCLC phenotypic transition and metastasis.

A prompt gamma neutron activation analysis (PGNAA) system based on a deuterium-deuterium (D-D) sealed neutron generator was designed using the MOCA code for explosive detection. The system is mainly composed of four parts: D-D sealed neutron generator, moderator, shielding, and Lutetium Yttrium OxyorthoSilicate (LYSO) scintillation detectors. Polyethylene (PE) was selected as the moderator and the optimal thickness was 7cm. Lead, PE, and boron-containing polyethylene were used as shielding materials. In the optimized model, the LYSO detector is used to measure eighteen materials, such as wood, melamine, glucose, and nylon, and so on. Firstly, the nitrogen characteristic peak of 10.8 MeV was analyzed to determine whether the material contained nitrogen. Then, the ratio of characteristic peak counts of C/O and O/N were calculated to distinguish explosives from nitrogen containing materials. Finally, dinitrobenzene, nitroglycerin, TNT, and ammonium nitrate can be separated from nitrogenous substances by a discriminant algorithm. The final device can be used to detect the chemical composition of the threat substances, and the maximum dose rate of the system meets the limits of international protection standards.

Glycosylphosphatidylinositol (GPI) anchor modification is a posttranslational modification of proteins that has been conserved in eukaryotes. The biosynthesis and transfer of GPI to proteins are carried out in the endoplasmic reticulum. Attachment of GPI to proteins is mediated by the GPI-transamidase (GPI-TA) complex, which recognizes and cleaves the C-terminal GPI attachment signal of precursor proteins. Then, GPI is transferred to the newly exposed C-terminus of the proteins. GPI-TA consists of five subunits: PIGK, GPAA1, PIGT, PIGS, and PIGU, and the absence of any subunit leads to the loss of activity. Here, we analyzed functionally important residues of the five subunits of GPI-TA by comparing conserved sequences among homologous proteins. In addition, we optimized the purification method for analyzing the structure of GPI-TA. Using purified GPI-TA, preliminary single particle images were obtained. Our results provide guidance for the structural and functional analysis of GPI-TA.

Background Type 2 cytokine Interleukin (IL)-13 regulates airway remodeling in asthma by acting on store-operated Ca2+ entry (SOCE) in airway smooth muscle cells. The underlying mechanisms of this regulating effect of IL-13 are not fully understood. Methods Bioinformatic analysis identified interactions of microRNA 93-5p with Orai1, the pore-forming molecule of SOCE, and long non-coding RNA H19 respectively. We investigated the role of H19/miR-93-5p/Orai1 axis in the regulation of proliferation and migration of in vitro cultured human bronchial smooth muscle cells (hBSMCs) induced by IL-13. Functional relevance of H19 in airway inflammation and airway remodeling was investigated in acute and chronic murine models of asthma. Results IL-13 dose-dependently increased the expression of H19 and Orai1 and decreased the expression of miR-93-5p in hBSMCs; H19 siRNA reversed IL-13-induced miR-93-5p and Orai1 expression, and the proliferation and migration of hBSMCs. IL-13-promoted expression of H19 and Orai1 was reduced by miR-93-5p mimic and enhanced by miR-93-5p inhibitor. IL-13-promoted hBSMCs proliferation was enhanced by miR-93-5p inhibitor but not changed by miR-93-5p mimic; whereas IL-13-promoted hBSMCs migration was enhanced by miR-93-5p inhibitor and reduced by miR-93-5p mimic. MiR-93-5p mimic enhanced the inhibiting effect of H19 siRNA on IL-13-induced Orai1 mRNA expression, whereas miR-93-5p inhibitor reversed the inhibiting effects of H19 siRNA on IL-13-induced H19 and Orai1 mRNA and protein expression. The inhibiting effect of H19 siRNA on IL-13-induced hBSMCs proliferation and migration was reversed by miR-93-5p inhibitor but not changed by miR-935p mimic. In the lungs of both asthma mice models, the expression of H19 and Orai1 was higher than in control mice. In acute asthma mice, H19 siRNA reduced Orai1 expression, inflammatory cell infiltration and goblet cell hyperplasia in the lungs, and IL-13 levels in the bronchoalveolar lavage fluid. In chronic asthma mice, H19 siRNA reduced Orai1 expression, inflammatory cell infiltration, goblet cell hyperplasia, collagen deposition and smooth muscle mass in the lungs, as well as IL-13 levels in the BALF. Conclusion IL-13 increases the proliferation and migration of airway smooth muscle cells by acting on H19/miR-93-5p/Orai1 axis, which also regulates airway inflammation and airway remodeling in murine models of asthma.

Immune-mediated, drug-induced hepatitis is a rare complication of halogenated volatile anesthetic administration. IL-4-regulated Th2-polarized reactions initiate this type and other types of hepatitis, while the mechanisms that regulate the severity remain elusive. IL-33 is an innate, IL-4-inducing, Th2-polarizing cytokine that has been detected in patients with liver failure and has been associated with upregulated ST2+Foxp3+CD4+CD25+ T cells; however, roles for IL-33 in drug-induced hepatitis are unclear. We investigated IL-33 in an anesthetic, immune-mediated hepatitis modeled in BALB/c, IL-33-/- and ST2-/- mice, as well as in patients with anesthetic hepatitis. The hepatic IL-33 and ST2 levels were elevated in BALB/c mice (p 0.05) with hepatitis, and anti-IL-33 diminished hepatitis (p 0.05) without reducing IL-33 levels. The complete absence of IL-33 reduced IL-10 (p 0.05) and ST2+Foxp3+CD4+CD25+ T cells (p 0.05), as well as reduced the overall survival (p 0.05), suggesting suppressive roles for IL-33 in anesthetic, immune-mediated hepatitis. All of the mice demonstrated similar levels of CD4+ T-cell proliferation following direct T-cell receptor stimulation, but we detected splenic IL-33 and ST2-negative Foxp3+CD4+CD25+ T cells in ST2-/- mice that developed less hepatitis than BALB/c mice (p 0.05), suggesting that ST2-negative Foxp3+CD4+CD25+ T cells reduced hepatitis. In patients, serum IL-33 and IPEX levels were correlated in controls (r

In the field of chemical safety, a named entity recognition (NER) model based on deep learning can mine valuable information from hazard and operability analysis (HAZOP) text, which can guide experts to carry out a new round of HAZOP analysis, help practitioners optimize the hidden dangers in the system, and be of great significance to improve the safety of the whole chemical system. However, due to the standardization and professionalism of chemical safety analysis text, it is difficult to improve the performance of traditional models. To solve this problem, in this study, an improved method based on active learning is proposed, and three novel sampling algorithms are designed, Variation of Token Entropy (VTE), HAZOP Confusion Entropy (HCE) and Amplification of Least Confidence (ALC), which improve the ability of the model to understand HAZOP text. In this method, a part of data is used to establish the initial model. The sampling algorithm is then used to select high-quality samples from the data set. Finally, these high-quality samples are used to retrain the whole model to obtain the final model. The experimental results show that the performance of the VTE, HCE, and ALC algorithms are better than that of random sampling algorithms. In addition, compared with other methods, the performance of the traditional model is improved effectively by the method proposed in this paper, which proves that the method is reliable and advanced.

BackgroundThis study was designed to investigate the validity of maximal oxygen consumption (VO2max) estimation through the Firstbeat fitness test (FFT) method when using submaximal rowing and running programs for well-trained athletes.MethodsWell-trained flatwater rowers (n = 45, 19.8 ± 3.0 years, 184 ± 8.7 cm, 76 ± 12.9 kg, and 58.7 ± 6.0 mL⋅kg–1⋅min–1) and paddlers (n = 45, 19.0 ± 2.5 years, 180 ± 7.7 cm, 74 ± 9.4 kg, and 59.9 ± 4.8 mL⋅kg–1⋅min–1) completed the FFT and maximal graded exercise test (GXT) programs of rowing and running, respectively. The estimated VO2max was calculated using the FFT system, and the measured VO2max was obtained from the GXT programs. Differences between the estimated and measured VO2max values were analyzed to assess the accuracy and agreement of the predictions. Equations from the previous study were also used to predict the VO2max in the submaximal programs to compare the accuracy of prediction with the FFT method.ResultsThe FFT method was in good agreement with the measured VO2max in both groups based on the intraclass correlation coefficients (>0.8). Additionally, the FFT method had considerable accuracy in VO2max estimation as the mean absolute percentage error (≤5.0%) and mean absolute error (–1⋅min–1) were fairly low. Furthermore, the FFT method seemed more accurate in the estimation of VO2max than previously reported equations, especially in the rowing test program.ConclusionThis study revealed that the FFT method provides a considerably accurate estimation of VO2max in well-trained athletes.

The Altun orogenic belt (AOB) was the site of complex subduction of oceanic crust and continental collision during the Paleozoic. The North Altun ophiolitic melange belt (NAB) is a one of the key tectonic units of the AOB and contains abundant subduction- and collision-related rocks. In this paper, we report the petrography, zircon U–Pb ages, geochemistry, and zircon Hf isotopes of the Bashikaogong S-type granitic rocks from the North Altun ophiolitic melange belt (NAB) to constrain their petrogenesis and tectonic setting. The granitic rocks consist of three types of granites, i.e., (1) gray, medium- to coarse-grained porphyritic granite; (2) gray, medium-grained granite; and (3) pink, medium- to coarse-grained granite. Zircon U–Pb dating yielded ages of 483–477, 458–453, and 447–445 Ma for type 1, type 2, and type s3 granites, respectively. All the three types of granites share similar strongly peraluminous (A/CNK > 1.1), contain muscovite mineral, have K2O/ Na2O ratios of > 1, and display negative zircon eHf(t) values, which are similar to typical S-type granites. On the basis of our data and results of previous studies, we infer that type 1 granites (483–477 Ma) are related to subduction of North Altun oceanic lithosphere, type 2 granites (458–453 Ma) are syn-collision granites related to continental collision between the Central Altun and Dunhuang Blocks, and type 3 granites (447–445 Ma) are from the late-collision stage. The Bashikaogong S-type granitic rocks recorded early Paleozoic subduction and collision in the North Altun region and were generated by the partial melting of the protolith of the metasedimentary Paleoproterozoic–Mesoproterozoic basement in the CAB due to the collapse of the overthickened subduction orogen.

BACKGROUND: Despite the rapid improvement of clinical science and imaging technology including computed tomography, the entity of negative surgical exploration in suspected gastrointestinal perforation (N-GIP) still exist. However, few studies have focused on this issue and most studies are case reports. We undertook this study to investigate the rates of N-GIP, and explore a set of possible preoperative predictors associated with N-GIP. METHODS: This was a retrospective study performed at the department of general surgery in our treatment center. All patients included were suspected gastrointestinal perforation (GIP) cases, aged 14 years and over, and underwent emergency surgery between 2009 and 2019. A predictive multivariable model of the presence of N-GIP was developed using logistic regression analysis. RESULTS: A total of 973 patients were identified and 30 (3.1%) were found to have no evidence of perforated gastrointestinal tract. The mean age of patients was 59.74 (range, 14–97) years, and 67.2 percent of the patients were males. The rates of N-GIP did not have a significant change over time (P=0.212 for trend). In multivariable analysis, absence of generalized peritonitis, duration of abdominal pain >19.6 hours, and neutrophil-to-lymphocyte ratio (NLR) 19.6 hours, and NLR

Small-cell lung cancer (SCLC) is a recalcitrant cancer characterized by high metastasis. However, the exact cell type contributing to metastasis remains elusive. Using a Rb1L/L/Trp53L/L mouse model, we identify the NCAMhiCD44lo/– subpopulation as the SCLC metastasizing cell (SMC), which is progressively transitioned from the non-metastasizing NCAMloCD44hi cell (non-SMC). Integrative chromatin accessibility and gene expression profiling studies reveal the important role of the SWI/SNF complex, and knockout of its central component, Brg1, significantly inhibits such phenotypic transition and metastasis. Mechanistically, TAZ is silenced by the SWI/SNF complex during SCLC malignant progression, and its knockdown promotes SMC transition and metastasis. Importantly, ectopic TAZ expression reversely drives SMC-to-non-SMC transition and alleviates metastasis. Single-cell RNA-sequencing analyses identify SMC as the dominant subpopulation in human SCLC metastasis, and immunostaining data show a positive correlation between TAZ and patient prognosis. These data uncover high SCLC plasticity and identify TAZ as the key molecular switch in orchestrating SCLC phenotypic transition and metastasis.

Lymphoma is a common and aggressive form of hematopoietic malignancies with diverse clinical and pathological features due to its heterogeneity. Although the current immunochemotherapeutic regimens improve clinical outcomes, many patients still display poor prognosis and frequent relapse. Epigenetic alterations contribute to the progression of lymphoma. DNA methylation and histone methylation are the most common epigenetic alterations and regulate the gene expression involved in lymphoma pathogenesis, including silencing of tumor suppressor genes or activation of proto-oncogenes. Dysregulation or mutation of genes related to DNA methylation, including DNMTs, TET2, IDH2, and genes related to histone methylation, including EZH2, KMT2D has been observed. Most of these alterations are associated with inferior outcomes of patients with diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), peripheral T-cell lymphoma (PTCL), and other subtypes of lymphoma. To overcome the pathogenetic consequence induced by aberrant DNA methylation and histone methylation, novel targeted drugs including azacitidine and decitabine have been gradually applied in practice to enhance the efficacy of current therapy and improve the prognosis of lymphoma patients. Investigating and targeting epigenetic mechanisms in lymphoma could be a key point of future research. Therefore, we mainly summarize the methylation alterations in lymphoma and their respective targeted therapies in this review.

Background: The impact of physical activity on immune response is a hot topic in exercise immunology, but studies involving asthmatic children are scarce. Our aims were to examine whether there were any differences in the level of physical activity and daily TV attendance, to assess its role on asthma control and immune responses to various immune stimulants. Methods: Weekly physical activity and daily television attendance were obtained from questionnaires at inclusion of the PreDicta study. PBMC cultures were stimulated with phytohemagglutinin (PHA), R848, poly I:C, and zymosan. A panel of cytokines was measured and quantified in cell culture supernatants using luminometric multiplex immunofluorescence beads-based assay. Results: Asthmatic preschoolers showed significantly more TV attendance than their healthy peers (58.6% vs. 41.5% 1–3 h daily and only 25.7% vs. 47.2% ≤1 h daily) and poor asthma control was associated with less frequent physical activity (PA) (75% no or occasional activity in uncontrolled vs. 20% in controlled asthma; 25% ≥3 times weekly vs. 62%). Asthmatics with increased PA exhibited elevated cytokine levels in response to polyclonal stimulants, suggesting a readiness of circulating immune cells for type 1, 2, and 17 cytokine release compared to subjects with low PA and high TV attendance. This may also represent a proinflammatory state in high PA asthmatic children. Low physical activity and high TV attendance were associated with a decrease in proinflammatory cytokines. Proinflammatory cytokines were correlating with each other in in vitro immune responses of asthmatic children, but not healthy controls, this correlation was more pronounced in children with sedentary behavior. Conclusion: Asthmatic children show more sedentary behavior than healthy subjects, while poor asthma control is associated with a substantial decrease in physical activity. Our results suggest that asthmatic children may profit from regular exercise, as elevated cytokine levels in stimulated conditions indicate an immune system prepared for responding strongly in case of different types of infections. However, it has to be considered that a hyperinflammatory state in high PA may not be beneficial in asthmatic children.

Environmental pH influences cell growth and differentiation. In the dimorphic yeast Yarrowia lipolytica, neutral-alkaline pH strongly induces the yeast-to-filament transition. However, the regulatory mechanism that governs alkaline pH-induced filamentation has been unclear. Here, we show that the pH-responsive transcription factor Y. lipolytica Rim101 (YlRim101) is a major regulator of alkaline-induced filamentation, since the deletion of YlRIM101 severely impaired filamentation at alkaline pH, whereas the constitutively active YlRIM1011-330 mutant mildly induced filamentation at acidic pH. YlRim101 controls the expression of the majority of alkaline-regulated cell wall protein genes. One of these, the cell surface glycosidase gene YlPHR1, plays a critical role in growth, cell wall function, and filamentation at alkaline pH. This finding suggests that YlRim101 promotes filamentation at alkaline pH via controlling the expression of these genes. We also show that, in addition to YlRim101, the Msn2/Msn4-like transcription factor Mhy1 is highly upregulated at alkaline pH and is essential for filamentation. However, unlike YlRim101, which specifically regulates alkaline-induced filamentation, Mhy1 regulates both alkaline- and glucose-induced filamentation, since the deletion of MHY1 abolished them both, whereas the overexpression of MHY1 induced strong filamentation irrespective of the pH or the presence of glucose. Finally, we show that YlRim101 and Mhy1 positively coregulate seven cell wall protein genes at alkaline pH, including YlPHR1 and five cell surface adhesin-like genes, three of which appear to promote filamentation. Together, these results reveal a conserved role of YlRim101 and a novel role of Mhy1 in the regulation of alkaline-induced filamentation in Y. lipolytica. IMPORTANCE The regulatory mechanism that governs pH-regulated filamentation is not clear in dimorphic fungi except in Candida albicans. Here, we investigated the regulation of alkaline pH-induced filamentation in Yarrowia lipolytica, a dimorphic yeast distantly related to C. albicans. Our results show that the transcription factor YlRim101 and the Msn2/Msn4-like transcription factor Mhy1 are the major regulators that promote filamentation at alkaline pH. They control the expression of a number of cell wall protein genes important for cell wall organization and filamentation. Our results suggest that the Rim101/PacC homologs play a conserved role in pH-regulated filamentation in dimorphic fungi.

Molecular heterogeneity of hepatobiliary tumor including intertumoral and intratumoral disparity always leads to drug resistance. Here, seven hepatobiliary tumor organoids are generated to explore heterogeneity and evolution via single‐cell RNA sequencing. HCC272 with high status of epithelia‐mesenchymal transition proves broad‐spectrum drug resistance. By examining the expression pattern of cancer stem cells markers (e.g., PROM1, CD44, and EPCAM), it is found that CD44 positive population may render drug resistance in HCC272. UMAP and pseudo‐time analysis identify the intratumoral heterogeneity and distinct evolutionary trajectories, of which catenin beta‐1 (CTNNB1), glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH), and nuclear paraspeckle assembly transcript 1 (NEAT1) advantage expression clusters are commonly shared across hepatobiliary organoids. CellphoneDB analysis further implies that metabolism advantage organoids with enrichment of hypoxia signal upregulate NEAT1 expression in CD44 subgroup and mediate drug resistance that relies on Jak‐STAT pathway. Moreover, metabolism advantage clusters shared in several organoids have similar characteristic genes (GAPDH, NDRG1 (N‐Myc downstream regulated 1), ALDOA, and CA9). The combination of GAPDH and NDRG1 is an independent risk factor and predictor for patient survival. This study delineates heterogeneity of hepatobiliary tumor organoids and proposes that the collaboration of intratumoral heterogenic subpopulations renders malignant phenotypes and drug resistance., The existence of inter‐ and intratumoral heterogeneity is the main cause for tumor drug resistance. Thus, extensive understanding of the underlying mechanism is necessary for developing potential strategy. This study here, for the first time, provides the new understanding for the role of tumor ecosystem involving cell expansion and drug response by applying scRNA‐seq method with tumor organoids.

Over 100 kinds of proteins are expressed as glycosylphosphatidylinositol (GPI)-anchored proteins (GPI-APs) on the cell surface in mammalian cells. GPI-APs possess unique properties in terms of their intracellular trafficking and association with lipid rafts. Although it is clear that GPI-APs play critical roles in various biological phenomena, it is poorly understood how the GPI moiety contributes to these mechanisms. More than 30 genes are involved in the correct biosynthesis of GPI-APs. We here constructed a cell library in which 32 genes involved in GPI biosynthesis were knocked out in human embryonic kidney 293 cells. Using the cell library, the surface expression and sensitivity to phosphatidylinositol-specific phospholipase C of GPI-APs were analyzed. Furthermore, we identified structural motifs of GPIs that are recognized by a GPI-binding toxin, aerolysin. The cell-based GPI-knockout library could be applied not only to basic researches, but also to applications and methodologies related to GPI-APs., Liu et al. construct a knock out cell library of genes involved in the biosynthesis of glycosylphosphatidylinositol (GPI)-anchored protein (GPI-APs) from HEK-293 cells and analyzed the surface expression and sensitivity to phosphatidylinositol-specific phospholipase C. Furthermore, the authors also identify structural motifs of GPIs that are recognized by a GPI-binding toxin, aerolysin.

As the manufacturing industry is facing increasingly serious environmental problems, because of which carbon tax policies are being implemented, choosing the optimum cutting parameters during the machining process is crucial for automobile panel dies in order to achieve synergistic minimization of the environment impact, product quality, and processing efficiency. This paper presents a processing task-based evaluation method to optimize the cutting parameters, considering the trade-off among carbon emissions, surface roughness, and processing time. Three objective models and their relationships with the cutting parameters were obtained through input–output, response surface, and theoretical analyses, respectively. Examples of cylindrical turning were applied to achieve a central composite design (CCD), and relative validation experiments were applied to evaluate the proposed method. The experiments were conducted on the CAK50135di lathe cutting of AISI 1045 steel, and NSGA-II was used to obtain the Pareto fronts of the three objectives. Based on the TOPSIS method, the Pareto solution set was ranked to find the optimal solution to evaluate and select the optimal cutting parameters. An S/N ratio analysis and contour plots were applied to analyze the influence of each decision variable on the optimization objective. Finally, the changing rules of a single factor for each objective were analyzed. The results demonstrate that the proposed method is effective in finding the trade-off among the three objectives and obtaining reasonable application ranges of the cutting parameters from Pareto fronts.

Parenteral vaccinations are not able to elicit effective systemic and gastrointestinal immune protection simultaneously because the lymphocytes are typically restricted to primed tissues. Although all-trans retinoic acid (atRA) was reported to trigger the gut-homing of immunocytes, the bioavailability and systemic immune responses remain limited for use in robust enteric vaccinations. Here, we show that co-delivery of atRA, CpG oligodeoxynucleotides (CpG), and antigens via engineered polymer/lipid nanoparticles (PLNPs) could exploit the amplifying function of draining lymph nodes (DLNs) for potent gut tropism and immune activations. After intramuscular injection, forming an immune-potentiated environment at the injection site, the PLNPs induced the designated transfer of primed dendritic cells (DCs) to the DLNs instead of the gastrointestinal tissues. Within the DLNs, the immune-potentiated environment markedly amplified the antigen presentation and homing receptor switch among immunocytes, which simultaneously stimulated the preferential dissipation of activated lymphocytes in the peripheral and gastrointestinal tissues, that is, exerted a DLN-amplifying effect. Compared with current atRA-containing formulations, the PLNPs not only boosted potent IgG secretions and T cell activations in the peripheral tissue but also provoked robust T cell homing and antigen-specific IgA levels in the gastrointestinal tracts in both ovalbumin and EV71 vaccinations. These data indicate that exploiting DLN amplification can stimulate potent systemic and gastrointestinal responses for more efficient enteric vaccinations.

Since the use of protein therapeutics is effective for treating intractable human diseases, the production of biologic therapeutic agents has dramatically increased over the past three decades. The Chinese hamster ovary (CHO) cell lines are the most commonly used host cell expression system for recombinant protein production. High productive and stable clonal cell lines for recombinant protein production have been established from the DHFR-deficient CHO cell using the dihydrofolate reductase/methotrexate (DHFR/MTX) selection methods. Human embryonic kidney 293 (HEK293) cells are alternative host cells widely used for protein production. In most case, however, the cells are used for the transient expression, and there is no gene amplification system in HEK293 cells. In this study, we established a DHFR-deficient HEK293 cell line for the high yield of recombinant proteins. We doubly knocked out DHFR and DHFR2 in the MAN1A1/A2/B1/C1-quadruple knockout HEK293 (QD-KO) cells, using the CRISPR/Cas9 system. The DHFR-deficient QD-KO cells were used to overexpress two proteins, lysosomal acid lipase and the constant fragment of human immunoglobulin G1 by the DHFR/MTX gene-amplification method. This method resulted in a dramatic increase in the two protein expressions in the DHFR-deficient QD-KO cells by increasing MTX concentration. Our system could be adopted in the production of several recombinant proteins including therapeutic proteins.

With growing concerns regarding the global environment, the industrial sector has played a significant role; that is, it is responsible for consuming large amount of energy and resources, while simultaneously producing wastes and carbon dioxide. A quantitative calculation of carbon emission in the turning process is presented in this paper. A generic carbon emission system boundary based on exergy balance is proposed first to avoid blurred system boundaries or the omission of elements. Then, a carbon emission model (iERWC) is formed by converting energy consumption (E), resource depletion (R) and waste generation (W) to equivalent carbon emissions (C) based on information flow (i), which effectively solves the problem of quantifying the impact of the machining process on the environment. Finally, the model is verified by experiments, and a simulation analysis is carried out. Additionally, the influence rule of processing parameters on carbon emissions is analyzed, and the cutting parameter that produces the lowest carbon emission is given.

In the normal heart, frequently used anesthetics such as isoflurane and propofol can reduce inotropy. However, the impact of these agents on the failing myocardium is unclear. Here, we examined whether and how isoflurane and propofol influence cardiac contractility in intact cardiac muscles from rats treated with monocrotaline to induce heart failure. We measured force and intracellular Ca(2+) ([Ca(2)(+)](i)) in trabeculae from the right ventricles of the rats in the absence or presence of propofol or isoflurane. At low to moderate concentrations, both propofol and isoflurane dose-dependently depressed cardiac force generation in failing trabeculae without altering [Ca(2+)](i). At high doses, propofol (but not isoflurane) also decreased amplitude of [Ca(2+)](i) transients. During steady-state activation, both propofol and isoflurane impaired maximal Ca(2+)-activated force (F(max)) while increasing the amount of [Ca(2+)](i) required for 50% of maximal activation (Ca(50)). These events occurred without apparent change in the Hill coefficient, suggesting no impairment of cooperativity. Exposing these same muscles to the anesthetics after fiber skinning resulted in a similar decrement in F(max) and rise in Ca(50) but no change in the myofibrillar ATPase-Ca(2+) relationship. Thus, our study demonstrates that challenging the failing myocardium with commonly used anesthetic agents such as propofol and isoflurane leads to reduced force development as a result of lowered myofilament responsiveness to Ca(2+). SIGNIFICANCE STATEMENT: Commonly used anesthetics such as isoflurane and propofol can impair myocardial contractility in subjects with heart failure by lowering myofilament responsiveness to Ca(2+). High doses of propofol can also reduce the overall amplitude of the intracellular Ca(2+) transient. These findings may have important implications for the safety and quality of intra- and perioperative care of patients with heart failure and other cardiac disorders.

The Altun orogenic belt in northwest China is part of the northern margin of the Tibetan Plateau. The North Altun ophiolitic melange belt is an important tectonic unit within the Altun orogenic belt that contains voluminous early Paleozoic granitoids. In this study, we report the petrological features, geochemical compositions, and zircon U Pb and Hf isotope data of three granitic plutons from the western segment of the North Altun ophiolitic melange belt. Zircon U Pb dating yields magmatic crystallization ages of 499, 493 and 496 Ma for samples of granodiorite, quartz diorite and syenogranite, respectively. The granitoids have metaluminous to weakly peraluminous and medium-K to high-K calc–alkaline characteristics and display relative enrichments in large ion lithophile elements (Rb, Th, U, K) and light rare earth elements (LREE) and relative depletions in Nb, Ta, Sr, P and Ti, suggesting an arc-related origin. The granodiorites (499 Ma) have positive zircon eHf(t) values ranging from +1.87 to +6.59 with two-stage Hf model ages (TDMC) of 1.05 to 1.35 Ga, implying that the granodiorites were derived from juvenile crust. The quartz diorites (493 Ma) have similar Hf isotopic characteristics to the granodiorites (eHf(t) = +2.59 to +6.04, TDMC = 1.08 to 1.30 Ga), indicating derivation from juvenile crust. The syenogranites (496 Ma) have high total REE and K2O contents, and low zircon eHf(t) values (−1.69 to +1.54), suggesting that they were derived mainly from juvenile crust mixed with ancient crustal materials. Combined with data from previous studies, we conclude that magmatism in the North Altun ophiolitic melange belt can be subdivided into three episodes: Episode 1 (520–470 Ma) granitoids are related to subduction; Episode 2 (460–425 Ma) granitoids formed in a continent–continent collisional setting; and Episode 3 (

Objective: To evaluate whether the combination of Uremic clearance granule with Alprostadil is superior to Alprostadil alone for CRF. Methods: Relevant RCTs were searched through March 2019. Data were analyzed by Stata 15.0. Results: Nine articles involving 726 patients were enrolled in this study. Meta-analysis showed that the total effective rate [OR = 3.68 (2.44, 5.55), P < 0.001], Scr [SMD = -2.34 (-3.49, -1.19), P < 0.001], BUN [SMD = -1.80 (-2.73, -0.87), P < 0.001], Ccr [SMD = 0.71 (0.44, 0.97), P < 0.001] were better in the experimental group. But there were no significant difference in UA, CysC, 24h-Upro and incidence of adverse reactions (all P > 0.05) between two groups. No serious adverse reactions were found. Conclusions: The effect of the integrated medicine on CRF was better than Alprostadil alone. Uremic clearance granule is safe and has no obvious adverse reactions.

The main objective of this paper is to propose a newly developed ocean Significant Wave Height (SWH) retrieval method from Envisat Advanced Synthetic Aperture Radar (ASAR) imagery. A series of wave mode imagery from January, April and May of 2011 are collocated with ERA-Interim reanalysis SWH data. Based on the matched datasets, a simplified empirical relationship between 22 types of SAR imagery parameters and SWH products is developed with the Genetic Algorithms Partial Least-Squares (GA-PLS) model. Two major features of the backscattering coefficient S10 and the frequency parameter S10 are chosen as the optimal training feature subset of SWH retrieval by using cross validation. In addition, we also present a comparison of the retrieval results of the simplified empirical relationship with the collocated ERA-Interim data. The results show that the assessment index of the correlation coefficient, the bias, the root-mean-square error of cross validation (RMSECV) and the scattering index (SI) are 0.78, 0.07 m, 0.76 m and 0.5, respectively. In addition, the comparison of the retrieved SWH data between our simplifying model and the Jason-2 radar altimeter data is proposed in our study. Moreover, we also make a comparison of the retrieval of SWH data between our developed model and the wellknown CWAVE_ENV model. The results show that satisfying retrieval results are acquired in the low-moderate sea state, but major bias appears in the high sea state, especially for SWH>5 m.

On the basis of our previous work defining the molecular rationale for combined targeting of the PI3K and AR pathways in PTEN loss prostate cancer, the first clinical trial was recently reported demonstrating a significant benefit for combination therapy in patients with metastatic prostate cancer. In this phase II trial, loss of PTEN was a biomarker predictive of response to combined AKT and AR inhibition. Given that PTEN loss prostate cancers are significantly enriched for ERG genomic rearrangements, we evaluated how the aberrant expression of ERG may impact response to PI3K/AR-targeted therapy. Here, we show that overexpression of ERG in the setting of Pten loss promotes resistance to combined PI3K and AR pathway inhibition with associated maintenance of AR target gene expression. Importantly, following AR knockout in the setting of ERG overexpression, there is maintenance of a subset of AR lineage–specific target genes, making AR dispensable in this context. This has important clinical implications as even in the setting of the androgen-regulated TMPRSS2:ERG genomic rearrangement, ERG expression is never abolished following AR inhibition and may allow for cell survival following AR (lineage)–targeted therapies.

The selectively direct conversion of methane to methanol on Cu-exchanged zeolites has attracted a lot of interest because of the abundant availability of methane. A detailed reaction mechanism of t...

We report our studies on the multiferroic orders in 0.5BiFeO3–0.5Bi0·5K0·5TiO3 (0.5BF–0.5BT) in this paper. The XRD pattern in the powdered form suggests a pseudocubic phase on average, with a lattice parameter of ∼3.9572 A. However, the bulk sample shows a coexistence of pseudocubic and tetragonal phases. The dielectric anomaly is relaxor-like, with the Burns temperature (TB) of ∼760 K. The remnant polarization (Pr, 4.02–4.25 μC/cm2) is nearly unchanged in the temperature range of 298 K and 398 K. In addition, the normalized strain coefficient (d × 33) increases on heating, from 100 pm/V at 298 K up to 240 pm/V at 453 K. On the other side, two anomalies were confirmed in the temperature dependence of the magnetic susceptibilities. The high-temperature anomaly is related to a transition between the antiferromagnetic and paramagnetic states around ∼143 K. The low-temperature anomaly around ∼5.5 K might hint the onset of the magnetic spin glass state. We believe that some common features should be present in (BixA1-x) (Fe0·5B0.5)O3 (A cations are not ferroelectric active while B cations are not magnetic active).

Organoid technology has been remarkably improved over the last decade. Various organoids have been derived from different types of tissues and recapitulate their organ-specific gene expression signatures, particular tissue spatial structures and functions of their original tissue. The patient-derived organoids (PDOs) have been used to elucidate crucial scientific questions, including the relationships between genetic/epigenetic alterations and drug responses, cell plasticity during disease progressions, and mechanisms of drug resistances. With the great expectations, PDOs will be widely used to facilitate the personalized medical decisions, which have the potential to profoundly improve patient outcomes. In this review, we will discuss the developmental details, current achievements, applications and challenges of organoid technology in precision cancer medicine.

Eu2+-activated phosphors are widely applied in lighting and display areas because of their good optical performance. In this paper, an excellent green-emitting zeolite-3A: 1.3 wt% Eu phosphor is prepared by a green and eco-friendly high-thermal reaction method without any reducing atmosphere or agents. Meanwhile, the reducing mechanism from Eu3+ ions to Eu2+ ions is investigated. The experiment results show that the morphology, crystal structure, and luminescent property are affected by sintering temperature. The resulting sample shows the broad excitation band is in the range of 310–450 nm and the peak of the broad emission band is located at 523 nm. Furthermore, zeolite-3A: 1.3 wt% Eu phosphor is encapsulated on a commercial UV-emitting chip to fabricate a purity green light-emitting diode (LED) with the Commission Internationale de L’Eclairage (CIE) color coordinates at (0.295, 0.537).

With rapid advances in flexible electronics and communication devices, flexible dielectric capacitors exhibiting high permittivity, low loss, and large electric-field tunability over a wide frequency range have attracted increasing attention. Here, a large-scale Ba0.67Sr0.33TiO3 (BST) dielectric thin film sandwiched between SrRuO3 (SRO) bottom electrode and Pt top electrode is fabricated on a flexible mica substrate. The mica/SRO/BST/Pt capacitor exhibits a dielectric constant (er′) of more than 1200, a loss tangent [tan(δ)] as low as 0.16, and a tunability of 67% at low frequencies around 10 kHz. Simultaneously, the capacitor can retain an er′ of 540 and a tan(δ) of 0.07 at microwave frequencies, e.g., 18.6 GHz. Moreover, even when the capacitor is bent to a small radius of 5 mm or undergoes 12 000 bending cycles (at 5 mm radius), almost no deterioration in er′, tan(δ), and tunability is observed. The excellent dielectricity and mechanical flexibility and durability endow the mica/SRO/BST/Pt capacitor wi...

Summary: Drug resistance is a significant hindrance to effective cancer treatment. Although resistance mechanisms of epidermal growth factor receptor (EGFR) mutant cancer cells to lethal EGFR tyrosine kinase inhibitors (TKI) treatment have been investigated intensively, how cancer cells orchestrate adaptive response under sublethal drug challenge remains largely unknown. Here, we find that 2-h sublethal TKI treatment elicits a transient drug-tolerant state in EGFR mutant lung cancer cells. Continuous sublethal treatment reinforces this tolerance and eventually establishes long-term TKI resistance. This adaptive process involves H3K9 demethylation-mediated upregulation of branched-chain amino acid aminotransferase 1 (BCAT1) and subsequent metabolic reprogramming, which promotes TKI resistance through attenuating reactive oxygen species (ROS) accumulation. Combination treatment with TKI- and ROS-inducing reagents overcomes this drug resistance in preclinical mouse models. Clinical information analyses support the correlation of BCAT1 expression with the EGFR TKI response. Our findings reveal the importance of BCAT1-engaged metabolism reprogramming in TKI resistance in lung cancer. : How cancer cells, with strong plasticity, orchestrate their adaptive response under sublethal drug exposure remains largely unknown. Wang et al. show that sublethal tyrosine kinase inhibitor (TKI) treatment elicits drug resistance in EGFR-mutant lung cancer cells through H3K9 demethylation-mediated reprogramming of branched-chain amino acid (BCAA) metabolism. Keywords: lung cancer, EGFR tyrosine kinase inhibitors, drug resistance, metabolic reprogramming, branched-chain amino acids, BCAT1

Energy conservation in manufacturing sector has received growing attention in an increasingly more carbon-restrained world due to the great concerns over climate change. The heavy-duty machine tool is usually the critical equipment in a factory, which consumes much more power than normal-sized machine tool but received less attention regarding energy saving. In this research, a generic power consumption model was developed from three hierarchies (i.e. system boundary definition, generic power consumption framework and detailed power consumption), which integrates the design parameters of the machine tool thus enabling the prediction of the power consumption even when the machine tool is physically unavailable. In addition, the relations between the power consumed in air-cutting and cutting states were also discussed, which can significantly simplify the study on machine tool power characteristics. The proposed model was verified by experiments and five power consumption characteristics of the tested heavy-duty machine tools were summarised. Additionally, four strategies for designing an energy-efficient machine tool and four tactics for using an existing machine tool more energy efficiently were proposed. This study represents part of a major comprehensive energy conservation research programme for heavy-duty machine tools, which aims to find solutions to improved energy efficiency for the manufacturing industry.

Due to the advantages of narrow energy distribution of plasmonic hot-electrons in Ag and the high density of states in the TiO2 conduction band, an Ag/TiO2 composite is considered to be an ideal combination to construct a plasmonic hot-electron photodetector with high detectivity and a high response speed. In this work, we fabricate a porous Ag/TiO2-Schottky-diode based plasmonic hot-electron photodetector. This detector shows a high detectivity of 9.8 × 1010 cmHz1/2/W and a fast response speed, with a rise and fall time of 112 μs and 24 μs, respectively, under 450 nm light illumination at zero bias voltage. In addition, the height of the Ag/TiO2 Schottky barrier can be decreased by removing the chemisorbed oxygen from the surface of TiO2 with ultraviolet light illumination, and as a result, the responsivity of the Ag/TiO2 plasmonic hot-electron photodetector at 450 nm can increase from 3.4 mA/W to 7.4 mA/W.

Right ventricular (RV) failure (RVF) has garnered significant attention in recent years because of its negative impact on clinical outcomes in patients with pulmonary hypertension (PH). PH triggers a series of events, including activation of several signaling pathways that regulate cell growth, metabolism, extracellular matrix remodeling, and energy production. These processes render the RV adaptive to PH. However, RVF develops when PH persists, accompanied by RV ischemia, alterations in substrate and mitochondrial energy metabolism, increased free oxygen radicals, increased cell loss, downregulation of adrenergic receptors, increased inflammation and fibrosis, and pathologic microRNAs. Diastolic dysfunction is also an integral part of RVF. Emerging non-invasive technologies such as molecular or metallic imaging, cardiac MRI, and ultrafast Doppler coronary flow mapping will be valuable tools to monitor RVF, especially the transition to RVF. Most PH therapies cannot treat RVF once it has occurred. A variety of therapies are available to treat acute and chronic RVF, but they are mainly supportive, and no effective therapy directly targets the failing RV. Therapies that target cell growth, cellular metabolism, oxidative stress, and myocyte regeneration are being tested preclinically. Future research should include establishing novel RVF models based on existing models, increasing use of human samples, creating human stem cell-based in vitro models, and characterizing alterations in cardiac excitation–contraction coupling during transition from adaptive RV to RVF. More successful strategies to manage RVF will likely be developed as we learn more about the transition from adaptive remodeling to maladaptive RVF in the future.

Two cases of medical malpractice for sciatic nerve injury caused by gluteal intramuscular injection in China are reported. Two children presented with foot drop indicative of sciatic nerve injury following gluteal intramuscular injections. The appraisal of whether there was medical negligence, the causal relationship between the patients’ nerve injuries and medical standard of care, and the causative potency were entrusted to us by the court. Based on each patient’s original medical history, imaging examination results, limb dysfunction and interviews with their relatives, there was a causal relationship between the children’s injuries and the medical treatment. The causative potency of medical negligence was complete effect in one case and main effect in the other case.

Background Glycosyltransferases are type II membrane proteins that are responsible for glycan modification of proteins and lipids, and localize to distinct cisternae in the Golgi apparatus. During cisternal maturation, retrograde trafficking helps maintain the steady-state localization of these enzymes in the sub-compartments of the Golgi. Methods To understand how glycosyltransferases are recycled in the late Golgi complex, we searched for genes that are essential for budding yeast cell growth and that encode proteins localized in endosomes and in the Golgi. We specifically analyzed the roles of Dop1 and its binding partner Neo1 in retaining Golgi-resident glycosyltransferases, in the late Golgi complex. Results Dop1 primarily localized to younger compartments of the trans-Golgi network (TGN) and seemed to cycle within the TGN. In contrast, Neo1, a P4-ATPase that interacts with Dop1, localized to the TGN. Abolition of DOP1 expression led to defects in the FM4-64 endocytic pathway. Dop1 and Neo1 were required for correct glycosylation of invertase, a secretory protein, at the Golgi. In DOP1-shutdown cells, Och1, a mannosyltransferase that is typically located in the cis-Golgi, mislocalized to the TGN. In addition, the function of multiple glycosyltransferases required for N- and O-glycosylation were impaired in DOP1-shutdown cells. Conclusions Our results indicate that Dop1 is involved in vesicular transport at the TGN, and is critical for retrieving glycosyltransferases from the TGN to the Golgi in yeast. General significance Golgi-resident glycosyltransferases recycling from the TGN to the Golgi is dependent on Dop1 and the P4-ATPase Neo1.

The Tagged Neutron Method (TNM) widely used in recent years has been applied to detect explosives to fight against terrorism. Because of the existence of a high-density (about 2.5 g/cm3) dry concrete wall, neutrons are moderated and γ -ray are attenuated and absorbed, which causes great interference to the detection of hidden explosives. A movable device has been designed for detection of explosive embedded in a concrete wall by using TNM technique. The experimental set-up was modeled and optimized with the MCNP5 Monte Carlo code. The γ -ray energy spectrum of graphite sample was firstly collected to calibrate the detection system and verify the validity of the MCNP5 model. Then γ -ray energy spectra of different samples under different situation are accumulated and compared with the simulation results. The samples of TNT and ammonium nitrate surrogate were placed behind a concrete wall with thickness of 10 cm, 15 cm and 20 cm, respectively. The comparison of the MCNP simulation results with experiment is in a good agreement. The results obtained provide a good reference for the subsequent data acquisition and analysis by using this model.

The growth mechanism of V-defects in GaN films was investigated. It was observed that the crystal faces of both the sidewall of a V-defect and the sidewall of the GaN film boundary belong to the same plane family of \{ {{{10\bar 11}}} \}, which suggests that the formation of the V-defect is a direct consequence of spontaneous growth like that of the boundary facet. However, the growth rate of the V-defect sidewall is much faster than that of the boundary facet when the V-defect is filling up, implying that lateral growth of \{ {{{10\bar 11}}} \} planes is not the direct cause of the change in size of V-defects. Since V-defects originate from dislocations, an idea was proposed to correlate the growth of V-defects with the presence of dislocations. Specifically, the change in size of the V-defect is determined by the growth rate around dislocations and the growth rate around dislocations is determined by the growth conditions.

Premise Evolutionary transitions among floral morphologies, many of which provide evidence for adaptation to novel pollinators, are common. Some trumpet-shaped flowers are among the largest flowers in angiosperms, occurring in different lineages. Our goal was to investigate the role of pollinators in the evolution of these flowers using Lilium. Methods We investigated floral traits and pollinators of L. primulinum var. ochraceum and L. brownii var. viridulum and reviewed reports of visitors to huge trumpet-shaped flowers. Using a published phylogeny of Lilium, we reconstructed ancestral floral morphological states in Lilium to elucidate the origins of trumpet-shaped lilies. Results Both lilies are largely self-incompatible and show floral syndromes indicative of hawkmoth pollination. The short trumpet-shaped lily can be pollinated by short-tongued ( 65 mm), while the huge trumpet-shaped lily can be pollinated by both. A literature review including 22 species of trumpet-shaped flowers suggests that their pollinator guilds commonly include both short- and long-tongued moths. A phylogenetic reconstruction indicates that trumpet-shaped lilies possibly have multiple origins from tepal-reflexed ancestors, at least six of which have evolved huge flowers (>50 mm). Conclusions Adaptation to short-tongued hawkmoths may have initiated the evolution of trumpet-shaped lilies. Huge trumpet-shaped lilies may have evolved as a response to selection by long-tongued hawkmoths, without excluding the short-tongued ones. This evolutionary pathway leads to a functionally more generalized pollination system instead of an increasingly specialized one and is not necessarily associated with pollinator shifts.

The asparagine (N)-linked Man9GlcNAc2 is required for glycoprotein folding and secretion. Understanding how its structure contributes to these functions has been stymied by our inability to produce this glycan as a homogenous structure of sufficient quantities for study. Here, we report the high yield chemoenzymatic synthesis of Man9GlcNAc2 and its biosynthetic intermediates by reconstituting the eukaryotic lipid-linked oligosaccharide (LLO) pathway. Endoplasmic reticulum mannosyltransferases (MTases) are expressed in E. coli and used for mannosylation of the dolichol mimic, phytanyl pyrophosphate GlcNAc2. These recombinant MTases recognize unique substrates and when combined, synthesize end products that precisely mimic those in vivo, demonstrating that ordered assembly of LLO is due to the strict enzyme substrate specificity. Indeed, non-physiological glycans are produced only when the luminal MTases are challenged with cytosolic substrates. Reconstitution of the LLO pathway to synthesize Man9GlcNAc2 in vitro provides an important tool for functional studies of the N-linked glycoprotein biosynthesis pathway., Attachment of the oligosaccharide Man9GlcNAc2 is required for glycoprotein folding and secretion but synthesizing this compound for structural and functional studies has remained challenging. Here, the authors achieve efficient Man9GlcNAc2 synthesis by reconstituting its biosynthetic pathway in vitro.

Cutting force and temperature are the key factors to be controlled during the orthopaedic surgery which could result in mechanical damage and necrosis of the bone tissue. Mechanistic modelling of the bone cutting process is expected to be an efficient method to understand and control these process challenges. However, due to the special structure and properties of the bone tissue (consist of osteon fibres and interstitial lamellae matrix), the conventional metal cutting models are not applicable in bone cutting process. This paper presents a novel cutting force and temperature mechanistic models for milling of bone. A cutting stress model of bone material was developed which takes into account its anisotropic characteristics based on the orthogonal cutting data. The cutting force coefficients are predicted incorporating the osteon orientation, tool geometry and edge effect with unified mechanics of cutting approach. Furthermore, a model of the induced cutting temperature based on heat flux developed during the process was proposed to predict the temperature distribution on bone cut surface. The experimental results showed a better consistency with the proposed model compared with the conventional Johnson-Cook model under different cutting conditions. A necrosis (potential cell injury from thermal effect) penetration depth was also proposed to evaluate the extent of thermal damage of bone tissue by the developed models. The proposed model can be used to assist the robotic surgery, to optimize the cutting parameters as well as to guide the orthopaedic tool design.

This paper presents a systematic study of Raman spectra of nanocomposite photonic crystals produced using opal matrices or anodic aluminum oxide and filled with various inorganic compounds. We compare Raman spectra of the starting inorganic compounds (K2Cr2O7, CrO3, and KIO3) and nanocomposite photonic crystals whose pores are filled with the inorganic substances. Our results open up the possibility of producing high-performance sensors of inorganic substances introduced into the pores of photonic crystals, based on analysis of enhanced Raman scattering in a nanocomposite photonic crystal.

To investigate the effect and mechanism of Xuefu Zhuyu Capsule (血府逐瘀胶囊, XZC) on pro-angiogenesis in the hindlimb ischemic model rats. A total of 100 Sprague Dawley rats were randomly divided into a model group, a regular-dose XZC group (0.48 g•kg-1•d-1) and a high-dose XZC group (0.96 g•kg-1•d-1) using random number table method. The model of hindlimb ischemic rats were made through femoral artery embolization with Bletilla microsphere agent. XZC were given on the first day after embolization surgery and lasted 5 days. Finally 72 models were obtained with 12 in each group for each time point. The lower ischemic limb was amputated on the third day after embolization surgery. Histopathological characters and the number of blood vessels of granulation tissues were observed at 36 and 48 h after amputation, respectively. The main genes were obtained from microarray analysis and were validated using real-time quantitative polymerase chain reaction. The vascular number of granulation tissues at both 36 and 48 h were characterized by new and fresh vessels. The number of angiogenesis in the high-dose XZC group at 36 and 48 h was greater compared with that in the regular-dose XZC and model groups (P