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Abstract Background Hypoxia-induced pulmonary hypertension (HPH) is a lethal cardiovascular disease with the characteristic of severe remodeling of pulmonary vascular. Although a large number of dysregulated mRNAs, lncRNAs, circRNAs, and miRNAs related to HPH have been identified from extensive studies, the competitive endogenous RNA (ceRNA) regulatory network in the pulmonary artery that responds to hypoxia remains largely unknown. Results Transcriptomic profiles in the pulmonary arteries of HPH rats were characterized through high-throughput RNA sequencing in this study. Through relatively strict screening, a set of differentially expressed RNAs (DERNAs) including 19 DEmRNAs, 8 DElncRNAs, 19 DEcircRNAs, and 23 DEmiRNAs were identified between HPH and normal rats. The DEmRNAs were further found to be involved in cell adhesion, axon guidance, PPAR signaling pathway, and calcium signaling pathway, suggesting their crucial role in HPH. Moreover, a hypoxia-induced ceRNA regulatory network in the pulmonary arteries of HPH rats was constructed according to the ceRNA hypothesis. More specifically, the ceRNA network was composed of 10 miRNAs as hub nodes, which might be sponged by 6 circRNAs and 7 lncRNAs, and directed the expression of 18 downstream target genes that might play important role in the progression of HPH. The expression patterns of selected DERNAs in the ceRNA network were then validated to be consistent with sequencing results in another three independent batches of HPH and normal control rats. The diagnostic effectiveness of several hub mRNAs in ceRNA network was further evaluated through investigating their expression profiles in patients with pulmonary artery hypertension (PAH) recorded in the Gene Expression Omnibus (GEO) dataset GSE117261. Dysregulated POSTN, LTBP2, SPP1, and LSAMP were observed in both the pulmonary arteries of HPH rats and lung tissues of PAH patients. Conclusions A ceRNA regulatory network in the pulmonary arteries of HPH rats was constructed, 10 hub miRNAs and their corresponding interacting lncRNAs, circRNAs, and mRNAs were identified. The expression patterns of selected DERNAs were further validated to be consistent with the sequencing result. POSTN, LTBP2, SPP1, and LSAMP were suggested to be potential diagnostic biomarkers and therapeutic targets for PAH.

Abstract. Objective:. Rubicon is an inhibitory interacting protein of the autophagy-related protein Uvrag. We previously showed that Rubicon deficiency promotes autophagic flux in vivo and that autophagy can degrade lipid droplets. This study aimed to investigate the effects of Rubicon deficiency on fasting-induced hepatic steatosis. Methods:. Two-month-old wild-type (WT) and Rubicon-deficient mice were subjected to feeding or fasting for 24 hours to induce hepatic steatosis. The distribution of liver lipid droplets was revealed by oil red O staining. Hepatic and plasma triglyceride, non-esterified fatty acid (NEFA), and cholesterol levels were detected using commercially available kits. Real-time reverse transcriptase-polymerase chain reaction was performed to analyze the mRNA expression of genes related to lipid metabolism in the liver. Western blot was conducted to assess autophagy-related protein levels in the liver. The animal experiments were approved by the Institutional Animal Care and Use Committee at Shanghai Jiao Tong University, China. Results:. We showed that under fasting conditions, Rubicon-deficient mice had more lipid droplets in the liver than WT controls. Consistent with these results, the hepatic triglyceride, NEFA, and cholesterol levels in fasted Rubicon-deficient mice were significantly higher than those of fasted WT controls. The levels of SREBP-1, a key regulator of lipid synthesis, were significantly lower in livers from fasted WT mice than those of fed WT mice. However, the decrease in SREBP-1 in fasted mice was attenuated by Rubicon deficiency. Western blot analysis demonstrated that the fasting-induced increase in autophagic flux was amplified by Rubicon deficiency. Finally, we showed that Rubicon deficiency in mice led to elevated plasma triglyceride and NEFA acid levels under fasting conditions. Conclusion:. Rubicon deficiency exacerbates fasting-induced hepatic steatosis in mice.

As the depth of mineral exploration increases, integrated geophysical methods are increasingly playing a crucial role in prospecting deep structures at the district scale. The Nanling-Xuancheng ore district is the eighth ore district in the middle-lower Yangtze metallogenic belt in China. To reveal the deep structure of the mining district, this study mainly focuses on regional high-precision gravity and magnetic data and integrates the interpretation of magnetotelluric and reflection seismic data from a key area. By using a 2.5D joint inversion method with prior information constraints, new insights into the deep structures, tectonic deformation, and magmatic activity are obtained. Structurally, the Nanling-Xuancheng ore district presents a structural pattern of “two uplifts and two depressions” composed of multi-level thrust-overturned and folds formed by Mesozoic depressions, which has a three-layer structure in the vertical direction (shallower than 10 km). Tectonically, the main faults in the study area trend NW, which intersect with NE-trending and EW-trending faults to form a branching structure from deep to shallow. The fault intersections provide pathways for magma intrusion. The distribution of deep-seated concealed magmatic rocks shows the characteristic pattern of “a primary magma source spawning multiple subsidiary intrusion”.

Atrioventricular septal defect (AVSD) is a deleterious subtype of congenital heart diseases (CHD) characterized by atrioventricular canal defect. The pathogenic genetic changes of AVSD remain elusive, particularly for copy number variation (CNV), a large segment variation of the genome, which is one of the major forms of genetic variants resulting in congenital heart diseases. In the present study, we recruited 150 AVSD cases and 100 healthy subjects as controls for whole exome sequencing (WES). We identified total 4255 rare CNVs using exon Hidden Markov model (XHMM) and screened rare CNVs by eliminating common CNVs based on controls and Database of Genomic Variants (DGV). Each patient contained at least 9 CNVs, and the CNV burden was prominently presented in chromosomes 19,22,21&16. Small CNVs (

Abstract Conversion of natural and secondary mixed forests to plantation monocultures can alter soil physical, chemical, and biological properties, which may lead to changes in ecosystem nitrogen (N) status by altering soil N availability. However, assessing and comparing N status between forest types within and across sites remain challenging because different indicators often yield contradicting conclusions. We developed a multiple‐indicator approach to evaluate N status across sites and between larch monocultures and mixed secondary forests paired at three sites (Daxinganling, Maoershan, and Qingyuan) in northeastern China. The selected indicators were foliar N content (%N), soil carbon‐to‐N ratio (C/N), soil inorganic N, foliar 15N natural abundance (δ15N), and δ15N of bulk soil, ammonium, and nitrate in the soil profiles. Our results, as shown mainly by C/N of soil organic layer and the fraction of nitrate in soil inorganic N, showed that N availability increased from north to south in the order Daxinganling

The interferon (IFN) response is the first line of defense against viral invasion and thus plays a central role in the regulation of the immune response. IFN-epsilon (IFN-ε) is a newly discovered type I IFN that does not require viral induction, unlike other type I IFNs. IFN-ε is constitutively expressed in epithelial cells and plays an important role in mucosal immunity. In this study, we evaluated the biological activity of the mink-IFN (MiIFN)-ε gene in prokaryotic cells. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used to evaluate IFN-ε expression in different mink tissues. MiIFN-ε was highly expressed in brain, lung, tracheal, kidney, intestinal, bladder, ovarian, and testis tissues. There was no significant difference in MiIFN-ε expression between female and male minks, except in the reproductive system. Expression of the small ubiquitin-like modifier (SUMO3)-MiIFN-ε fusion gene was induced by isopropylβ-d-thiogalactoside, and MiIFN-ε was collected after SUMO-specific protease digestion. We tested the antiviral activity of MiIFN-ε against vesicular stomatitis virus (VSV) in epithelial cells of feline kidney 81 (F81). We used qRT-PCR to analyze the expression of several IFN-stimulated genes (ISGs), including ISG15, 2′-5′ oligoadenylate synthetase (2′-5′OAS1), and myxovirus resistance protein 1 (Mx1). Recombinant IFN-ε induced high ISG expression in F81 cells. Compared with those in the cell control group, expressions of ISG15, Mx1, and 2′-5′ OAS1 in the VSV-GFP control, IFN-ε, and MiIFN-ε-inhibited VSV-GFP groups were significantly increased. Compared with those in the VSV-GFP control group, expressions of ISG15 and 2′-5′ OAS1 in the IFN-ε and MiIFN-ε-inhibited VSV-GFP groups were significantly increased, and the differences were highly significant (p < 0.0001). IFN-ε played an indirect antiviral role. These findings lay the foundation for detailed investigation of IFN-ε in the future.

Coalbed methane (CBM) resources are abundant in the Baoan blocks of the Xinjing coalfield, China; their exploration and development remain in infancy. Suitable development blocks can be selected by evaluating their development potential; that of the No. 8 and 15 CBM systems in the Baoan blocks was evaluated using three quantitative evaluation methods via existing geological data: multi-level fuzzy mathematics (MLFM), technique for order preference by similarity to ideal solution (TOPSIS), and principal component analysis (PCA). The results demonstrated that the CBM development potential in the central part of the Baoan blocks is low and unbecoming. The favorable areas of the No. 8 coal seam are distributed in the northeast and northwest, and that of No. 15 in the northwest and southwest Differences exist in quantitative selection; with limited data, the evaluation of the TOPSIS method is considerably different from actuality, and is unbecoming for evaluating the development potential.

Around 11% of all known gene lesions causing human genetic diseases are nonsense mutations that introduce a premature stop codon (PTC) into the protein-coding gene sequence. Drug-induced PTC readthrough is a promising therapeutic strategy for treating hereditary diseases caused by nonsense mutations. To date, it has been found that more than 50 small-molecular compounds can promote PTC readthrough, known as translational readthrough-inducing drugs (TRIDs), and can be divided into two major categories: aminoglycosides and non-aminoglycosides. This review summarizes the pharmacodynamics and clinical application potential of the main TRIDs discovered so far, especially some newly discovered TRIDs in the past decade. The discovery of these TRIDs brings hope for treating nonsense mutations in various genetic diseases. Further research is still needed to deeply understand the mechanism of eukaryotic cell termination and drug-induced PTC readthrough so that patients can achieve the greatest benefit from the various TRID treatments.

BackgroundImmune checkpoint inhibitors (ICIs) combined with chemotherapy have been widely employed to improve the outcome of gastric cancer patients. In the present study, the impact of posttreatment growth hormone (GH) levels on the treatment efficacy of ICIs for advanced gastric cancer (AGC) patients was assessed.MethodsSeventy-five AGC patients treated with anti-PD-1 antibodies at The Fourth Hospital of Hebei Medical University were involved. We divided AGC patients into two groups as high-GH group and low-GH group based on the GH level. Immunotherapy efficacy was assessed in terms of objective response rate, disease control rate (DCR), progression-free survival (PFS), and overall survival (OS) based on the National Comprehensive Cancer Network Guidelines. The enumeration data were compared by χ2 test or Fisher’s exact test. Survival curves were drawn by the Kaplan–Meier method, and comparisons between the curves were made using the log-rank test. Multivariate survival analysis was performed using a Cox proportional hazards model.ResultsThe higher GH levels were associated with a lower DCR of ICIs with a DCR of 30.0% in the high-GH group and 53.3% in the low-GH group (P = 0.046). The subsequent univariate analysis showed that a high GH level was associated with both shorter PFS (P = 0.016) and shorter OS at the borderline statistical level (P = 0.052) in AGC patients treated with ICIs. Cox model analysis also proved that the GH level was an independent risk factor for the outcome of AGC patients (PFS: P = 0.013, HR, 2.424, 95% CI, 1.202–4.890; OS: P = 0.014, HR, 3.301, 95% CI, 1.279–8.519).ConclusionsThe post-treatment GH level might be a predictor for ICIs treatment in AGC patients.

Hereditary Spastic Paraplegia (HSP) is considered to be one of the common neurodegenerative diseases with marked genetic heterogeneity. Recently, the mutations in ubiquitin-associated protein 1 (UBAP1) have been described in patients with HSP, known as spastic paraplegias 80 (SPG80). Here, we reported a Chinese HSP family presenting a frameshift mutation in the UBAP1 gene leading to complex HSP. Their clinical features encompassed spastic paraparetic gait, exaggerated patellar tendon reflexes, bilateral Babinski signs, and hyperactive Achilles tendon reflex. The proband also had severe urinary incontinence and a dermoid cyst at the lumbar 4–5 spinal cord, which rarely occurs in HSP patients. Following whole-exome sequencing, a novel heterozygous mutation (c.437dupG, NM_016,525) was identified in the UBAP1 that segregated with the family’s phenotype and resulted in truncating UBAP1 protein (p.Ser146ArgfsTer13). Moreover, we reviewed the genotypes of UBAP1 and the phenotypic variability in 90 HSP patients reported in the literature. We found that the age of onset in UBAP1-related patients was juvenile, and there were population differences in the age of onset. The main complications were lower extremity spasticity, hyperreflexia, and the Babinski sign. Exon 4 of UBAP1 was identified as a mutation hotspot region. Our study expands the knowledge of UBAP1 mutations, which will aid in HSP patient counseling. Further molecular biological research is needed to explore the genotype-phenotype correlations of UBAP1-related HSP.

Phenoloxidase (PO)–catalyzed melanization is a vital immune response in insects for defense against pathogen infection. This process is mediated by clip domain serine proteases and regulated by members of the serpin superfamily. We here revealed that the infection of Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) significantly inhibited the PO activity in Ostrinia furnacalis hemolymph and induced the expression of O. furnacalis serpin–4. Addition of recombinant serpin-4 protein to O. furnacalis hemolymph resulted in a great increase of AcMNPV copies. Serpin-4 significantly suppressed the PO activity and the amidase activity in cleaving colorimetric substrate IEARpNA (IEARase activity) of hemolymph. Further experiments indicated it formed covalent complexes with three serine proteases (SP1, SP13 and SP105) and prevented them from cleaving their cognate downstream proteases in vitro. Altogether, O. furnacalis melanization restricted AcMNPV replication and serpin-4 facilitated AcMNPV infection by inhibiting serine proteases, SP1, SP13, and SP105 which were all involved in the melanization response.

Aim of study: To explore the different patterns of fine-root traits by elucidating changes in their chemistries and morphologies in Chinese fir plantations along an altitudinal gradient. Area of study: National Mazongling Nature Reserve (Anhui Province). Material and methods: Soil and fine roots (≤ 2 mm) samples were extracted from three soil layers (0-10 cm, 10-20 cm, and 20-30 cm) at four altitudes (750 m, 850 m, 1000 m, and 1150 m), after which their nutrient concentrations and morphological traits, respectively, were quantified. We employed mixed model ANOVA to test the effects of altitude, soil layer, and their interactions on the characteristics of soil and fine roots. The relationships between the functional traits of fine roots and climate, soil and stand structures were evaluated by the standard major axis regression and the structural equation model. Main results: The chemical traits of fine roots were higher at medium altitudes (which decreased significantly with the soil layer). In contrast the morphological traits of fine roots did not change significantly. In chemical traits, both altitude, organic matter components, and soil total phosphorus (TP) exerted dominant effects on fine-root N, and both altitude and soil TP exerted dominant effects on fine-root P. However, in morphological traits, we found that altitude and soil C:N were crucial impact factors. Research highlights: Fine roots might preferentially adjust their chemical traits rather than morphological traits to facilitate higher root efficiencies in response to variable environmental conditions.

Dync1li1, a subunit of cytoplasmic dynein 1, is reported to play important roles in intracellular retrograde transport in many tissues. However, the roles of Dync1li1 in the mammalian cochlea remain uninvestigated. Here we first studied the expression pattern of Dync1li1 in the mouse cochlea and found that Dync1li1 is highly expressed in hair cells (HCs) in both neonatal and adult mice cochlea. Next, we used Dync1li1 knockout (KO) mice to investigate its effects on hearing and found that deletion of Dync1li1 leads to early onset of progressive HC loss via apoptosis and to subsequent hearing loss. Further studies revealed that loss of Dync1li1 destabilizes dynein and alters the normal function of dynein. In addition, Dync1li1 KO results in a thinner Golgi apparatus and the accumulation of LC3+ autophagic vacuoles, which triggers HC apoptosis. We also knocked down Dync1li1 in the OC1 cells and found that the number of autophagosomes were significantly increased while the number of autolysosomes were decreased, which suggested that Dync1li1 knockdown leads to impaired transportation of autophagosomes to lysosomes and therefore the accumulation of autophagosomes results in HC apoptosis. Our findings demonstrate that Dync1li1 plays important roles in HC survival through the regulation of autophagosome transportation.

Forests worldwide store large quantities of carbon (C), particularly in soils as soil organic C (SOC). In northeastern China, two dominant forest types, secondary mixed forest (MF) and larch plantation forest (LF), cover extensive areas. However, we lack an understanding of the patterns and the mechanisms of SOC storage and stabilization in MF and LF, especially in deep soil layers. This research aims to illustrate the vertical distribution and mineral protection of SOC over soil depth; we also used δ13C values of soil fractions to evaluate SOC stability. Samples from the surface litter (Oi), organic layer (Oa+e), and 0–40 cm mineral soils were collected from both MF and LF plots. We used two different methods to separate bulk soils into distinguished fractions: (1) macro- and micro-aggregates and the non-aggregated fraction, and (2) particulate organic matter (POM) and mineral-associated organic matter (MAOM). The C concentrations, C stocks, and δ13C of all soil fractions were determined. Our findings were as follows: (1) SOC was mainly stored in mineral soils and was 13.6% lower in LF (8609 ± 1180 g C m−2) than MF (9969 ± 2084 g C m−2). (2) In both MF and LF, the SOC stock was mainly stored in aggregates (averaged 92.7%); macroaggregates dominated in the surface layers (Oa+e layer and 0–10 cm) but microaggregates dominated in the deep layers (10–20 cm and 20–40 cm). In mineral soils, MAOM was the dominant fraction of the C stock (averaged 81.6%). (3) The proportion of C distributed in microaggregates and MAOM increased from Oa+e to the 20–40 cm layer. (4) The C/N ratios and δ13C values of MAOM were smaller and heavier compared to those of POM. Our study demonstrated that in both forests, aggregate formation and mineral association predominantly contributed to SOC storage, and large stocks of SOC were distributed in the deep soil. The increasing proportion of SOC in microaggregates and MAOM along the soil depth, most likely derived from microbial turnover and microbial necromass, influenced SOC stability in both forest types.

Listeria monocytogenes is an important foodborne pathogen, and is ubiquitously distributed in the natural environment. Cattle and sheep, as natural hosts, can transmit L. monocytogenes to related meat and dairy products. In this study, the prevalence, distribution, and transmission characteristics of Listeria were analysed by investigating 5214 samples of cattle and sheep in farm and slaughtering environments in China. A low contamination incidence of L. monocytogenes (0.5%, 20/4430) was observed in farm environment, but there was a high contamination incidence in slaughtering environment (9.4%, 74/784). The incidence of L. innocua in cattle and sheep farm and slaughtering environments is more common and significantly higher (9.7%, 508/5214) than that of L. monocytogenes (1.8%, 94/5214). The distinct molecular and genetic characteristics of Listeria by PFGE and MLST indicated that L. monocytogenes and L. innocua were gradually transmitted from the farm and slaughtering environments to end products, such as beef and mutton along the slaughtering chain. The ST7, ST9, ST91, and ST155 found in our study were associated with the human listeriosis cases in China. In addition, the findings of virulence markers (inlC, inlJ, LIPI-3, LIPI-4, and ECIII) concerned with the pathogenesis of human listeriosis and antibiotics resistance of L. monocytogenes in this study implies a potential public health risk. This study fills the gap in the epidemiology of beef cattle and sheep that carry Listeria in farm and slaughtering environments in major cattle and sheep producing areas in China.

Abstract Assessment of nitrogen (N) saturation of forests is critical to the evaluation of the manner in which ecosystems will respond to current and future global changes such as N deposition. However, quantifying N saturation remains a challenge. We developed a conceptual model of N saturation stages in forest ecosystems based on (1) a hypothetical relative rate of ammonification, nitrification, and denitrification, (2) concentrations of ammonium and nitrate in the soil, and (3) 15N enrichment pattern of bulk soil N, ammonium, and nitrate in the soil profile. We tested the hypotheses using data from forests located at five sites across eastern Asia. The fraction of nitrate in total inorganic N indicated that the sites represent an N saturation gradient with one boreal forest at stage 1 (least saturated), three temperate forests at stage 2, and one tropical forest at stage 3 (most saturated). The δ15N of bulk soil N increased from topsoil to subsoil more sharply at N‐limited sites than at the N‐rich sites along the N deposition gradient. We also found distinct 15N enrichment patterns of bulk soil N, ammonium, and nitrate in the soil profile across the study sites. At stage 1, nitrate was more 15N‐depleted than ammonium only in the organic soil horizon, indicating limited nitrification, while the 15N depletion of nitrate to ammonium was observed in the deeper mineral soil at stages 2 and 3. Furthermore, ammonium was more 15N‐depleted than bulk soil N at stages 1 and 2 but more 15N‐enriched than bulk soil N at stage 3. Our study suggests that soil profile patterns of δ15N of bulk soil N, ammonium, and nitrate provide information about the relative rates of mineralization, nitrification, and denitrification and thus can be an additional measure of N saturation of forest ecosystems across broad environmental gradients.

Abstract Background The biosynthesis of high value-added compounds using metabolically engineered strains has received wide attention in recent years. Myo-inositol (inositol), an important compound in the pharmaceutics, cosmetics and food industries, is usually produced from phytate via a harsh set of chemical reactions. Recombinant Escherichia coli strains have been constructed by metabolic engineering strategies to produce inositol, but with a low yield. The proper distribution of carbon flux between cell growth and inositol production is a major challenge for constructing an efficient inositol-synthesis pathway in bacteria. Construction of metabolically engineered E. coli strains with high stoichiometric yield of inositol is desirable. Results In the present study, we designed an inositol-synthesis pathway from glucose with a theoretical stoichiometric yield of 1 mol inositol/mol glucose. Recombinant E. coli strains with high stoichiometric yield (> 0.7 mol inositol/mol glucose) were obtained. Inositol was successfully biosynthesized after introducing two crucial enzymes: inositol-3-phosphate synthase (IPS) from Trypanosoma brucei, and inositol monophosphatase (IMP) from E. coli. Based on starting strains E. coli BW25113 (wild-type) and SG104 (ΔptsG::glk, ΔgalR::zglf, ΔpoxB::acs), a series of engineered strains for inositol production was constructed by deleting the key genes pgi, pfkA and pykF. Plasmid-based expression systems for IPS and IMP were optimized, and expression of the gene zwf was regulated to enhance the stoichiometric yield of inositol. The highest stoichiometric yield (0.96 mol inositol/mol glucose) was achieved from recombinant strain R15 (SG104, Δpgi, Δpgm, and RBSL5-zwf). Strain R04 (SG104 and Δpgi) reached high-density in a 1-L fermenter when using glucose and glycerol as a mixed carbon source. In scaled-up fed-batch bioconversion in situ using strain R04, 0.82 mol inositol/mol glucose was produced within 23 h, corresponding to a titer of 106.3 g/L (590.5 mM) inositol. Conclusions The biosynthesis of inositol from glucose in recombinant E. coli was optimized by metabolic engineering strategies. The metabolically engineered E. coli strains represent a promising method for future inositol production. This study provides an essential reference to obtain a suitable distribution of carbon flux between glycolysis and inositol synthesis.

Analyzing the characteristics of rock brittleness in low-permeability mudstone and shale (MS) formations is imperative for efficient hydraulic fracturing stimulation. Rock brittleness depends on the mineral composition, organic matter abundance, and bedding structure. Based on the MS from Shahejie Formation mineral composition (clay mineral, felsic mineral, and calcareous mineral contents), total organic content, and bedding structure (laminated, laminar, and massive), six types of lithofacies were identified: clay-rich MS, felsic-rich MS, calcareous-rich MS, clay MS, felsic MS, and calcareous MS. The quartz, feldspar, calcite, and dolomite of the Shahejie Formation are brittle minerals. Consequently, lithofacies with high felsic and calcareous mineral contents are more brittle. In addition, laminated and laminar MS are also conducive to hydraulic fracturing. Therefore, laminated, organic-rich, and calcareous-rich MS are the dominant lithofacies for hydraulic fracturing in the Shahejie Formation. The lithofacies and brittleness index were predicted by the response characteristics between mineral compositions and logging curves. The 3521–3552 m section of well B11x is dominated by calcareous-rich MS with developed laminae, representing a favorable section for hydraulic fracturing. Fragile minerals and oil are widely developed in the lower part of the lower 1st member of the Shahejie Formation (Es1L) in the southwestern part of Zhaohuangzhuang-Suning, where hydraulic fracturing can be used to increase shale oil production.

ABSTRACT Analysis of nitrogen isotope fractionation effects is useful for tracing biogeochemical nitrogen cycle processes. Nitrification can cause large nitrogen isotope effects through the enzymatic oxidation of ammonia (NH3) via nitrite (NO2−) to nitrate (NO3−) (15εNH4+→NO2- and 15εNO2-→NO3-). The isotope effects of ammonia-oxidizing bacteria (AOB) and archaea (AOA) and of nitrite-oxidizing bacteria (NOB) have been analyzed previously. Here, we studied the nitrogen isotope effects of the complete ammonia oxidizer (comammox) Nitrospira inopinata that oxidizes NH3 to NO3−. At high ammonium (NH4+) availability (1 mM) and pH between 6.5 and 8.5, its 15εNH4+→NO2- ranged from −33.1 to −27.1‰ based on substrate consumption (residual substrate isotopic composition) and −35.5 to −31.2‰ based on product formation (cumulative product isotopic composition), while the 15εNO2-→NO3- ranged from 6.5 to 11.1‰ based on substrate consumption. These values resemble isotope effects of AOB and AOA and of NOB in the genus Nitrospira, suggesting the absence of fundamental mechanistic differences between key enzymes for ammonia and nitrite oxidation in comammox and canonical nitrifiers. However, ambient pH and initial NH4+ concentrations influenced the isotope effects in N. inopinata. The 15εNH4+→NO2- based on product formation was smaller at pH 6.5 (−31.2‰) compared to pH 7.5 (−35.5‰) and pH 8.5 (−34.9‰), while 15εNO2-→NO3- was smaller at pH 8.5 (6.5‰) compared to pH 7.5 (8.8‰) and pH 6.5 (11.1‰). Isotopic fractionation via 15εNH4+→NO2- and 15εNO2-→NO3- was smaller at 0.1 mM NH4+ compared to 0.5 to 1.0 mM NH4+. Environmental factors, such as pH and NH4+ availability, therefore need to be considered when using isotope effects in 15N isotope fractionation models of nitrification. IMPORTANCE Nitrification is an important nitrogen cycle process in terrestrial and aquatic environments. The discovery of comammox has changed the view that canonical AOA, AOB, and NOB are the only chemolithoautotrophic organisms catalyzing nitrification. However, the contribution of comammox to nitrification in environmental and technical systems is far from being completely understood. This study revealed that, despite a phylogenetically distinct enzymatic repertoire for ammonia oxidation, nitrogen isotope effects of 15εNH4+→NO2- and 15εNO2-→NO3- in comammox do not differ significantly from those of canonical nitrifiers. Thus, nitrogen isotope effects are not suitable indicators to decipher the contribution of comammox to nitrification in environmental samples. Moreover, this is the first systematic study showing that the ambient pH and NH4+ concentration influence the isotope effects of nitrifiers. Hence, these key parameters should be considered in comparative analyses of isotope effects of nitrifiers across different growth conditions and environmental samples.

An electronic nose (E-Nose) and gas chromatography-ion mobility spectrometry (GC-IMS) were used to analyze the volatile flavor compounds (VFCs) of the enzymatic hydrolysate of Lanmaoa asiatica and its Maillard reaction products (MRPs). E-Nose sensors have strong response signals to sulfide, nitrogen oxides, alcohols, and aldehyde ketone, and the aroma profile was increased after the Maillard reaction (MR). According to GC-IMS, A total of 84 known compounds were identified. Aldehydes, ketones and alcohols are the main VFCs. After MR, the concentrations of some alcohols decreased, and the concentration of pyrazines and ketones increased. Principal component analysis (PCA) and similarity analysis showed that the enzymatic hydrolysate and MRPs were different and could be effectively distinguished. In conclusion, this study clarified the changes in VFCs before and after the MR. The results can provide a theoretical basis for the quality control and flavor changes during the processing of Lanmaoa asiatica and provide a new method for flavor analysis of edible mushrooms and their products.

The super elongation complex (SEC) has been reported to play a key role in the proliferation and differentiation of mouse embryonic stem cells. However, the expression pattern and function of the SEC in the inner ear has not been investigated. Here, we studied the inner ear expression pattern of three key SEC components, AFF1, AFF4, and ELL3, and found that these three proteins are all expressed in both cochlear hair cells (HCs)and supporting cells (SCs). We also cultured Lgr5+ inner ear progenitors in vitro for sphere-forming assays and differentiation assays in the presence of the SEC inhibitor flavopiridol. We found that flavopiridol treatment decreased the proliferation ability of Lgr5+ progenitors, while the differentiation ability of Lgr5+ progenitors was not affected. Our results suggest that the SEC might play important roles in regulating inner ear progenitors and thus regulating HC regeneration. Therefore, it will be very meaningful to further investigate the detailed roles of the SEC signaling pathway in the inner ear in vivo in order to develop effective treatments for sensorineural hearing loss.

Diabetes is an increasingly common disease that poses an immense challenge to public health. Hyperglycemia is also a common complication in clinical patients in the intensive care unit, increasing the rate of infection and mortality. The accurate and real-time prediction of blood glucose concentrations after each short-acting insulin injection has great clinical significance and is the basis of all intelligent blood glucose control systems. Most previous prediction methods require long-term continuous blood glucose records from specific patients to train the prediction models, resulting in these methods not being used in clinical practice. In this study, we construct 13 deep neural networks with different architectures to atomically predict blood glucose concentrations after arbitrary independent insulin injections without requiring continuous historical records of any patient. Using our proposed models, the best root mean square error of the prediction results reaches 15.82 mg/dL, and 99.5% of the predictions are clinically acceptable, which is more accurate than previously proposed blood glucose prediction methods. Through the re-validation of the models, we demonstrate the clinical practicability and universal accuracy of our proposed prediction method.

The dairy industry occupies an important position in the development of China's animal husbandry. It is a prominently developed industry in the animal husbandry industry and an important symbol of the country's agricultural development. The development of the dairy industry is of extremely important significance for promoting the transformation and upgrading of China's animal husbandry industry, increasing farmers' income, and enhancing the health quality of the entire nation. Therefore, the study aimed to explore the genetic effect of selective functional groups on milk production traits in Chinese Holstein cows to find the genes that caused mutations, and to study the identification strategies of the major genes that determined milk production traits. DNA was extracted from the frozen sperm genomes of 1,109 Holstein cows, and coagulated DNA from their blood was extracted. The mixed gene pool was sequenced, and different mutation sites were screened to classify genes. The collected data were statistically processed to analyze the correlation between genes. Haploid association analysis showed that all 35 Single Nucleotide Polymorphisms (SNP) and 3 indels were strongly associated with at least one trait, and 1, 1, 3, and 1 haplotype blocks were found in DDIT3, RPL23A, SESN2, and NR4A1 genes after continuous disequilibrium analysis. Correlation analysis showed that some haplotypes were correlated with lactation traits in dairy cows. The analysis of JASPAR software showed that 11 SNP sites and 3 binding sites had different degrees of changes, suggesting that this site would affect the transcriptional activity of transcription factor. The genetic effects of four candidate functional genes were further verified, and the four genes were identified as significantly associated with milk yield and milk composition traits in Chinese Holstein cow for the first time. [ABSTRACT FROM AUTHOR]

Abstract Background Acetyl-CoA is an important metabolic intermediate and serves as an acetylation precursor for the biosynthesis of various value-added acetyl-chemicals. Acetyl-CoA can be produced from glucose, acetate, or fatty acids via metabolic pathways in Escherichia coli. Although glucose is an efficient carbon source for acetyl-CoA production, the pathway from acetate to acetyl-CoA is the shortest and fatty acids can produce acetyl-CoA through fatty acid oxidation along with abundant NADH and FADH2. In this study, metabolically engineered E. coli strains for efficiently supplying acetyl-CoA from glucose, acetate, and fatty acid were constructed and applied in one-step biosynthesis of N-acetylglutamate (NAG) from glutamate and acetyl-CoA. Results A metabolically engineered E. coli strain for NAG production was constructed by overexpressing N-acetylglutamate synthase from Kitasatospora setae in E. coli BW25113 with argB and argA knockout. The strain was further engineered to utilize glucose, acetate, and fatty acid to produce acetyl-CoA. When glucose was used as a carbon source, the combined mutants of ∆ptsG::glk, ∆galR::zglf, ∆poxB::acs, ∆ldhA, and ∆pta were more efficient for supplying acetyl-CoA. The acetyl-CoA synthetase (ACS) pathway and acetate kinase-phosphate acetyltransferase (ACK-PTA) pathway from acetate to acetyl-CoA were investigated, and the ACK-PTA pathway showed to be more efficient for supplying acetyl-CoA. When fatty acid was used as a carbon source, acetyl-CoA supply was improved by deletion of fadR and constitutive expression of fadD under the strong promoter CPA1. Comparison of acetyl-CoA supply from glucose, acetate and palmitic acid revealed that a higher conversion rate of glutamate (98.2%) and productivity (an average of 6.25 mmol/L/h) were obtained when using glucose as a carbon source. The results also demonstrated the great potential of acetate and fatty acid to supply acetyl-CoA, as the molar conversion rate of glutamate was more than 80%. Conclusions Metabolically engineered E. coli strains were developed for NAG production. The metabolic pathways of acetyl-CoA from glucose, acetate, or fatty acid were optimized for efficient acetyl-CoA supply to enhance NAG production. The metabolic strategies for efficient acetyl-CoA supply used in this study can be exploited for other chemicals that use acetyl-CoA as a precursor or when acetylation is involved.

Long term stability is a major barrier for the commercialization of halide perovskite solar cells. Here Wu et al. demonstrate that a chemically inert and structural impermeability bismuth electrode interlayer greatly increases the stability of unencapsulated perovskite solar cells under harsh conditions.

Improving the entrepreneurial performance of farmers is conducive to increasing their income, reducing poverty, and securing livelihoods. In the quest for sustainable livelihoods, non-cognitive skills are becoming increasingly significant. Based on data from the 2018 Chinese Family Panel Study, this paper uses the ‘Big Five’ personality traits and a cross-sectional regression model to construct an indicator system to analyze farmers’ non-cognitive skills empirically and determine how these skills affect entrepreneurial performance. The results are as follows: (1) non-cognitive skills that significantly affect farmers’ entrepreneurial performance are, in order of influence, openness, extroversion, conscientiousness, and agreeableness. After considering endogeneity and substitution-explained variables, the above conclusions are still robust. (2) Heterogeneity analysis finds that farmers over 40 and those who start their businesses in cities have higher returns on non-cognitive entrepreneurship. (3) The paper confirms that non-cognitive skills improve farmers’ entrepreneurial performance through human and social capital effects. Therefore, entrepreneurs should consciously improve their non-cognitive skills by cultivating an enterprising and innovative spirit and social skills. The government’s entrepreneurship training for farmers should also focus on setting up courses in non-cognitive skills development to enhance farmers’ entrepreneurial literacy and skills.

A theoretical model has been proposed to predict the horizontal dimensional accuracy of selective laser melting (SLM) in this paper. It is found that the horizontal dimensional deviation of SLM consists of two parts. The first part is related to the mode of track filling and the track width. The effect of heat accumulation on track width is also taken into account in this model. The second part is the solidification shrinkage which is dominated by the temperature history of the tracks of a layer when the material and dimension size are fixed. Then, the SLMed Ti6Al4V parts fabricated to verify this theoretical model has been performed in this paper too. The results show that the predicted and experimental results has a good correspondence. At last, less than 20μm dimension deviation of SLMed thin-wall samples was achieved by pre-compensation using this theoretical model. Keywords: Theoretical model, Horizontal dimensional accuracy, Selective laser melting, Track width, Solidification shrinkage

In view of the widely existed two-dimensional friction problems in engineering, an oscillator model which considers the coupling of and direction is established by introducing oblique springs. With the concept of the friction force direction angle being defined to determine the components of the static and kinetic friction force vector, and considering that stick-slip motion may occur when the oscillator is vibrating, a method for analyzing stick-slip motion of a two-dimensional coupled dry friction oscillator is proposed, and complex stick-slip transition boundaries of the two-dimensional coupled dry friction oscillator are given. Based on an exponential-type dynamic friction model, numerical simulation is carried out for stick-slip motion of the two-dimensional coupled dry friction oscillator, with the simulation results and the law of the system solution changing with excitation frequency and phase angle under three different working conditions at x direction and y direction, namely the same excitation frequency and phase angle, the same excitation frequency and different phase angle, and different excitation frequency and phase angle, are provided. Numerical results indicate that transition of stick-slip state may occur in the motion of the two-dimensional coupled dry friction oscillator; in comparison with the first two conditions, when frequencies and phase angle of and direction of the excitation are not equal, trajectory of the mass is a more complicated plane curve, the system can have multiple stick-slip transitions in one cycle at the same time; with the same frequency of and direction, the response of the oscillator has no fractional frequencies and is periodic by altering the frequency and phase angle. The proposed method can provide reference for further study of dynamics and motion stability of a two-dimensional coupled dry friction oscillator.

Recent advances in neuroscience indicate that analysis of bio-signals such as rest state electroencephalogram (EEG) and eye-tracking data can provide more reliable evaluation of children autism spectrum disorder (ASD) than traditional methods of behavior measurement relying on scales do. However, the effectiveness of the new approaches still lags behind the increasing requirement in clinical or educational practices as the “bio-marker” information carried by the bio-signal of a single-modality is likely insufficient or distorted. This study proposes an approach to joint analysis of EEG and eye-tracking for children ASD evaluation. The approach focuses on deep fusion of the features in two modalities as no explicit correlations between the original bio-signals are available, which also limits the performance of existing methods along this direction. First, the synchronization measures, information entropy, and time-frequency features of the multi-channel EEG are derived. Then a random forest applies to the eye-tracking recordings of the same subjects to single out the most significant features. A graph convolutional network (GCN) model then naturally fuses the two group of features to differentiate the children with ASD from the typically developed (TD) subjects. Experiments have been carried out on the two types of the bio-signals collected from 42 children (21 ASD and 21 TD subjects, 3–6 years old). The results indicate that (1) the proposed approach can achieve an accuracy of 95% in ASD detection, and (2) strong correlations exist between the two bio-signals collected even asynchronously, in particular the EEG synchronization against the face related/joint attentions in terms of covariance.

Striatin-interacting protein 1 (Strip1) is a core component of the striatin interacting phosphatase and kinase (STRIPAK) complex, which is involved in embryogenesis and development, circadian rhythms, type 2 diabetes, and cancer progression. However, the expression and role of Strip1 in the mammalian cochlea remains unclear. Here we studied the expression and function of Strip1 in the mouse cochlea by using Strip1 knockout mice. We first found that the mRNA and protein expression of Strip1 increases as mice age starting from postnatal day (P) 3 and reaches its highest expression level at P30 and that the expression of Strip1 can be detected by immunofluorescent staining starting from P14 only in cochlear HCs, and not in supporting cells (SCs). Next, we crossed Strip1 heterozygous knockout (Strip +/−) mice to obtain Strip1 homozygous knockout (Strip1−/−) mice for studying the role of Strip1 in cochlear HCs. However, no Strip1−/− mice were obtained and the ratio of Strip +/− to Strip1+/+ mice per litter was about 2:1, which suggested that homozygous Strip1 knockout is embryonic lethal. We measured hearing function and counted the HC number in P30 and P60 Strip +/− mice and found that they had normal hearing ability and HC numbers compared to Strip1+/+ mice. Our study suggested that Strip1 probably play important roles in HC development and maturation, which needs further study in the future.

In this paper, we compare several non-tight degree evaluation methods i.e., Boura and Canteaut’s formula, Carlet’s formula as well as Liu’s numeric mapping and division property proposed by Todo, and hope to find the best one from these methodsfor practical applications. Specifically, for the substitution-permutation-network (SPN) ciphers, we first deeply explore the relationships between division property of an Sbox and its algebraic properties (e.g., the algebraic degree of its inverse). Based on these findings, we can prove theoretically that division property is never worse than Boura and Canteaut’s and Carlet’s formulas, and we also experimentally verified that the division property can indeed give a better bound than the latter two methods. In addition, for the nonlinear feedback shift registers (NFSR) based ciphers, according to the propagation of division property and the core idea of numeric mapping, we give a strict proof that the estimated degree using division property is never greater than that of numeric mapping. Moreover, our experimental results on Trivium and Kreyvium indicate the division property actually derives a much better bound than the numeric mapping. To the best of our knowledge, this is the first time to give a formal discussion on the relationships between division property and other degree evaluation methods, and we present the first theoretical proof and give the experimental verification to illustrate that division property is the optimal one among these methods in terms of the accuracy of the upper bounds on algebraic degree.

Humoral immune responses in animals are often tightly controlled by regulated proteolysis. This proteolysis is exerted by extracellular protease cascades, whose activation culminates in the proteolytic cleavage of key immune proteins and enzymes. A model for such immune system regulation is the melanization reaction in insects, where the activation of prophenoxidase (proPO) leads to the rapid formation of eumelanin on the surface of foreign entities such as parasites, bacteria and fungi. ProPO activation is tightly regulated by a network of so-called clip domain serine proteases, their proteolytically inactive homologs, and their serpin inhibitors. In Anopheles gambiae, the major malaria vector in sub-Saharan Africa, manipulation of this protease network affects resistance to a wide range of microorganisms, as well as host survival. However, thus far, our understanding of the molecular make-up and regulation of the protease network in mosquitoes is limited. Here, we report the function of the clip domain serine protease CLIPB10 in this network, using a combination of genetic and biochemical assays. CLIPB10 knockdown partially reversed melanotic tumor formation induced by Serpin 2 silencing in the absence of infection. CLIPB10 was also partially required for the melanization of ookinete stages of the rodent malaria parasite Plasmodium berghei in a refractory mosquito genetic background. Recombinant serpin 2 protein, a key inhibitor of the proPO activation cascade in An. gambiae, formed a SDS-stable protein complex with activated recombinant CLIPB10, and efficiently inhibited CLIPB10 activity in vitro at a stoichiometry of 1.89:1. Recombinant activated CLIPB10 increased PO activity in Manduca sexta hemolymph ex vivo, and directly activated purified M. sexta proPO in vitro. Taken together, these data identify CLIPB10 as the second protease with prophenoloxidase-activating function in An. gambiae, in addition to the previously described CLIPB9, suggesting functional redundancy in the protease network that controls melanization. In addition, our data suggest that tissue melanization and humoral melanization of parasites are at least partially mediated by the same proteases.