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Shale gas is a green, low-carbon and clean unconventional natural gas energy. China is rich in shale gas resources. Accelerating shale gas exploration and development is of great practical significance to improve China's energy structure and achieve the goal of “2030 carbon peak neutralization and 2060 carbon neutralization”. The Ordovician Wufeng Formation to Silurian Longmaxi Formation in Sichuan Basin is the key strata for shale gas exploration and development in China. Based on the experimental analysis of shale gas in Wufeng Formation to Longmaxi Formation in Weiyuan, Changning, Zhaotong, Fuling and Weirong areas of Sichuan Basin, this paper systematically analyzes the geochemical characteristics of shale gas in Wufeng Formation to Longmaxi Formation, discusses the genesis of shale gas, the causes of hydrocarbon isotope inversion and the source of shale gas, and looks forward to the exploration prospects of shale gas resources in the basin. It has important guiding significances for shale gas exploration and development in Sichuan Basin and even in China. The results show that: (1) The shale gas of Wufeng-Longmaxi formations is a typical dry gas, and the hydrocarbon isotopes are distributed in negative sequence. The carbon isotopes of alkane gas in Changning, Zhaotong and Fuling areas are heavier and have a higher degree of thermal evolution than those in Weiyuan and Weirong areas, and the rare gas is of crustal origin. (2) Shale gas of Wufeng-Longmaxi formations is a thermogenic oil type gas at high-over mature stage, which is mainly the mixture of crude oil cracking gas and kerogen cracking gas. The hydrocarbon isotope inversion of alkane gas is mainly caused by the mixing of crude oil cracking gas and kerogen cracking gas at high-over mature stage, the exchange between formation water and methane at high evolution stage and so on. (3) The carbon isotopic values of methane in shale gas of Wufeng-Longmaxi formations match the carbon isotopic values of kerogen in mudstone of Longmaxi Formation of Lower Silurian, which conforms to the law of carbon isotopic fractionation δ13Ckerogen>δ13Coil>δ13Calkane gas. (4) The total amount of marine, marine continental transitional and continental facies shale gas resources in Sichuan Basin is about 41.5 × 1012 m3, with broad exploration prospects in resource potential.

The Taodonggou group of Middle Permian is an important source rock in Taibei sag of Turpan-Hami basin. Due to its deep burial, drilling has only been revealed in recent years. Based on organic petrology and organic geochemistry experiments, this paper studies the organic petrology, organic geochemistry, sedimentary environment, and hydrocarbon generation potential of source rocks in Taibei sag, Turpan-Hami basin, and reveals the influence of the sedimentary environment on the organic matter abundance of source rocks. The results are as follows: (1) The organic matter of the Middle Permian source rocks in Taibei sag of Turpan-Hami basin is mainly sapropelite and exinite. The vitrinite is mainly vitrodetrinite, and the exinite is mainly lamalginiite. (2) The total organic carbon content value is 0.55–6.08 wt %, and the average value is 2.58 wt %. The PG value ranges from 0.78 mg HC/g to 30.86 mg HC/g, and the average value is 4.88 mg HC/g. Chloroform asphalt “A” is 0.046–0.8767 wt %, and the average value is 0.285 wt %. The types of organic matter are mainly III and II–III, and the Ro value is 0.628–1.49 wt % (average = 0.988 wt %). The Tmax distribution is 329–465 °C. The average temperature is 434.7 °C, which is in the mature stage (oil window stage). The Middle Permian source rocks are mainly very good to excellent source rocks with a good hydrocarbon generation potential. (3) The source rocks are deposited in a semihumid and semiarid climate. Organic matter is input as a mixed source. The early and late stages is dominated by terrestrial higher plants. The middle stage is dominated by lower aquatic organisms, and the sedimentary environment consists of weak reduction and weak oxidation environments. (4) In the study area, the abundance of organic matter has a weak negative correlation with CPI and a positive correlation with Pr/Ph and ∑C21–/∑C22+. Under the coaction of paleoclimate, organic matter input, and redox environment, the enrichment model of organic matter with high productivity and weak oxidation environment characteristics can also form excellent source rocks. This study is of great significance and provides theoretical guidance for the exploration of deep oil and gas resources.

Porcine reproductive and respiratory syndrome virus (PRRSV) continues to cause significant economic loss worldwide and remains a serious threat to the pork industry. Currently, vaccination strategies provide limited protection against PRRSV infection, and consequently, new antiviral strategies are urgently required. Andrographolide (Andro) and its derivative potassium dehydrographolide succinate (PDS) have been used clinically in China and other Asian countries as therapies for inflammation-related diseases, including bacterial and viral infections, for decades. Here, we demonstrate that Andro and PDS exhibit robust activity against PRRSV replication in Marc-145 cells and primary porcine alveolar macrophages (PAMs). The two compounds exhibited broad-spectrum inhibitory activities in vitro against clinically circulating type 2 PRRSV GD-HD, XH-GD, and NADC30-like HNhx strains in China. The EC(50) values of Andro against three tested PRRSV strain infections in Marc-145 cells ranged from 11.7 to 15.3 μmol/L, with selectivity indexes ranging from 8.3 to 10.8, while the EC(50) values of PDS ranged from 57.1 to 85.4 μmol/L, with selectivity indexes ranging from 344 to 515. Mechanistically, the anti-PRRSV activity of the two compounds is closely associated with their potent suppression on NF-κB activation and enhanced oxidative stress induced by PRRSV infection. Further mechanistic investigations revealed that PDS, but not Andro, is able to directly interact with PRRSV particles. Taken together, our findings suggest that Andro and PDS are promising PRRSV inhibitors in vitro and deserves further in vivo studies in swine. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12250-021-00455-y.

Breakthroughs have been made for the natural gas exploration of the Middle Devonian and the Middle Permian formations in the Sichuan Basin in recent years. Nevertheless, its natural gas source is still not clear, which then affects the next exploration deployment decision. A comprehensive study is conducted on the geochemical characteristics of the Middle Devonian and the Middle Permian natural gas, the biomarkers of reservoir bitumen, and the source rocks. The results show that these natural gases are secondary cracking dry gas that includes more than 86% of methane content, a small amount of ethane, propane, and a non-hydrocarbon gas (e.g., nitrogen, carbon dioxide, and hydrogen sulfide). The natural gas maturity varies slightly in different regions. The carbon and hydrogen isotope values of natural gas are as follows: δ13C1 ranges from −35.7‰ to −27.3‰, δ13C2 ranges from −38.7‰ to −26.6‰, δ13C3 ranges from −37‰ to −26.5‰, δ2HCH4 ranges from −141‰ to −138‰, and δ2HC2H6 ranges from −164‰ to −112‰. The isotope values indicate that the genetic type of natural gas is the sapropelic type and the mixture type is dominated by sapropelic type. The difference between natural gas values of δ13C1, δ13C2, and δ2HCH4 in different regions is related to the contribution ratio of source rocks from different ages. The gases of the Middle Devonian and the Middle Permian of the Shuangyushi structure in Northwestern Sichuan, the Middle Permian of Southwestern Sichuan, and the Middle Permian of Paleo-uplift in Central Sichuan are mainly derived from the Cambrian Qiongzhusi Formation and the Middle-Lower Permian source rocks. The Middle-Upper Permian natural gases of the volcanic reservoir in Southwestern Sichuan is mainly from the Qiongzhusi source rock. The Middle Permian natural gases of the Hewanchang structure in Northwestern Sichuan, Eastern Sichuan, and Southern Sichuan are mainly derived from the Silurian Longmaxi source rock. The research results have important guiding significance for developing exploration and deployment solutions for natural gas of the Middle Devonian and the Middle Permian formation in the Sichuan Basin.

The purpose of this research was to investigate whether copper (Cu) exposure could induce apoptosis via endoplasmic reticulum stress (ERS) in skeletal muscle of broilers. A total of 240 one-day-old chickens were randomly divided into four groups by free access; the diets are as follows: control diet (Cu 11 mg/kg, control group) and high level of Cu diets (Cu 110 mg/kg, group I; Cu 220 mg/kg, group II; Cu 330 mg/kg, group III). The skeletal muscle tissues were collected on day 49 for further examination. The content of Cu, histopathology, and the expression levels of the genes and proteins related to ERS and apoptosis were detected. Results showed that the Cu levels in skeletal muscle were increased in a dose-dependent manner. Meanwhile, the spaces between the muscle fibers were wider with the increase of Cu content, and the myolysis was observed in group III. Besides, the mRNA expression levels of GRP78, GRP94, eIF2α, ATF6, XBP1, CHOP, Caspase-12, and Caspase3 were markedly increased in treated groups compared with control group, and the protein expression levels of GRP78, Caspase3, Active-Caspase3 and JNK were significantly elevated with the increase of dietary Cu. In summary, these findings suggested that Cu could induce apoptosis through ERS in skeletal muscle of broilers.

Fluorine being a well-known and essential element for normal physiological functions of tissues of different organisms is frequently used for growth and development of body. The mechanisms of adverse and injurious impacts of fluoride are not clear and still are under debate. Therefore, this study was executed to ascertain the potential mechanisms of sodium fluoride in liver tissues of ducks. For this purpose, a total of 14 ducks were randomly divided and kept in two groups including control group and sodium fluoride treated group. The ducks in control group were fed with normal diet while the ducks in other group were exposed to sodium fluoride (750 mg/kg) for 28 days. The results showed that exposure to sodium fluoride induced deleterious effects in different liver tissues of ducks. The results indicated that mRNA levels of Cas-3, Cas-9, p53, Apaf-1, Bax and Cyt-c were increased in treated ducks with significantly higher mRNA level of Cas-9 and lower levels of the mRNA level of Bcl-2 as compared to untreated control group (P < 0.01). The results showed that protein expression levels of Bax and p53 were increased while protein expression level of Bcl-2 was reduced in treated ducks. No difference was observed in protein expression level of Cas-3 between treated and untreated ducks. The results of this study suggest that sodium fluoride damages the normal structure of liver and induces abnormal process of apoptosis in hepatocyte, which provide a new idea for elucidating the mechanisms of sodium fluoride induced hepatotoxicity in ducks.

Copper (Cu), one of the heavy metals, is far beyond the carrying capacity of the environment with Cu mining, industrial wastewater discharging and the use of Cu-containing pesticides. Intaking excess Cu can cause toxic effects on liver, kidney, heart, but few studies report Cu toxicity on brain tissue. It is noteworthy that most toxicity tests are based on rodent models, but large mammals chosen as animal models has no reported. To explore the relationship of the Cu toxicity and mitochondria-mediated apoptosis on hypothalamus in pigs, the content of Cu, histomorphology, mitochondrial related indicators, apoptosis, and AMPK-mTOR signaling pathway were detected. Results showed that Cu could accumulate in hypothalamus and lead to mitochondrial dysfunction, evidenced by the decrease of ATP production, activities of respiratory chain complex I-IV, and mitochondrial respiratory function in Cu-treated groups. Additionally, the genes and proteins expression of Bax, Caspase-3, Cytc in treatment group were higher than control group. Furthermore, the protein level of p-AMPK was enhanced significantly and p-mTOR was declined, which manifested that AMPK-mTOR signaling pathway was activated in Cu-treated groups. In conclusion, this study illuminated that the accumulation of Cu could cause mitochondrial dysfunction, induce mitochondria-mediated apoptosis and activate AMPK-mTOR pathway in hypothalamus.

Colorectal cancer (CRC) is the fourth most lethal cancer in the world. Heat shock protein 60 (HSP60), a mitochondrial chaperone that maintains mitochondrial proteostasis, is highly expressed in tumors compared with in paracancerous tissues, suggesting that high HSP60 expression benefits tumor growth. To determine the effects of HSP60 expression on tumor progression, stable HSP60-knockdown HCT116 cells were constructed in the present study, revealing that knockdown of HSP60 inhibited cell proliferation. Proteomic analysis demonstrated that mitochondrial proteins were downregulated, indicating that knockdown of HSP60 disrupted mitochondrial homeostasis. Metabolomic analysis demonstrated that cellular adenine levels were >30-fold higher in HSP60-knockdown cells than in control cells. It was further confirmed that elevated adenine activated the AMPK signaling pathway, which inhibited mTOR-regulated protein synthesis to slow down cell proliferation. Overall, the current results provide a valuable resource for understanding mitochondrial function in CRC, suggesting that HSP60 may be a potential target for CRC intervention.

Ovarian cancer (OC) is the most lethal gynecological carcinoma due to the lack of diagnostic markers and effective drug targets. Discovery of new therapeutic targets in OC to improve the treatment outcome is urgently needed. We performed proteomic analysis of OC specimens and the paired normal tissues and revealed that proteins associated with mitochondrial proteostasis and protein translation were highly expressed in ovarian tumor tissues, indicating that mitochondria are required for tumor progression of OC. Heat shock protein 60 (HSP60), an important mitochondrial chaperone, was upregulated in ovarian tumors. HSP60 silencing significantly attenuated growth of OC cells in both cells and mice xenografts. Proteomic analysis revealed that HSP60 silencing downregulated proteins involved in mitochondrial functions and protein synthesis. Metabolomic analysis revealed that HSP60 silencing resulted in a more than 100-fold increase in cellular adenine levels, leading to increased adenosine monophosphate and an activated AMPK pathway, and consequently reduced mTORC1-mediated S6K and 4EBP1 phosphorylation to inhibit protein synthesis that suppressed the proliferation of OC cells. These results suggest that HSP60 knockdown breaks mitochondrial proteostasis, and inactivates the mTOR pathway to inhibit OC progression, suggesting that HSP60 is a potential therapeutic target for OC treatment.

Due to Van der Waals or electrostatic interactions of surfactants active to all kinds of minerals, in low-rank coal (LRC) flotation, surfactants could both adsorb on the organic and mineral matter of LRC, thus increasing the hydrophobicity of them and resulting in the increase of clean coal yield as well as the ash content. In this work, density functional theory (DFT) calculations were clarified the adsorption mechanism of surfactant (C12EO, lauryl polyoxyethylene ether) on the LRC model molecules and kaolinite surfaces (the representative of organic and mineral matter in LRC). The DFT results demonstrate that the interaction strength between C12EO and LRC model molecules is stronger than that of kaolinite. Based on the calculation results, ultrasonic treatment was selected to expand the wettability difference between organic and mineral matter that adsorbed C12EO. The microcalorimetry results show that the wetting heat of organic matter that is adsorbed C12EO is unchanged basically after 30 s ultrasonic treatment. However, there is an obvious increase trend for the wetting heat of kaolinite. Flotation experiments of artificial mixed samples proved that ultrasonic treatment is useful in decreasing the extent of ash content in LRC flotation, thus improving the quality of clean coal in LRC flotation.

The aim of this study was to investigate the effects of excessive copper (Cu)-induced cytotoxicity on oxidative stress and mitochondrial apoptosis in chicken hepatocytes. Chicken hepatocytes were cultured in medium in the absence and presence of copper sulfate (CuSO4) (10, 50, 100 μM), in N-acetyl-L-cysteine (NAC) (1 mM), and the combination of CuSO4 and NAC for 24 h. Morphologic observation and function, reactive oxygen species (ROS) level, antioxidant indices, nitric oxide (NO) content, mitochondrial membrane potential (MMP), and apoptosis-related mRNA and protein levels were determined. These results indicated that excessive Cu could induce release of intracellular lactate dehydrogenase (LDH), aspartate aminotransferase (AST), and alanine aminotransferase (ALT); increase levels of ROS, superoxide dismutase (SOD), malondialdehyde (MDA), catalase (CAT), lipid peroxidation (LPO), and NO; decrease glutathione (GSH) content and MMP; upregulated Bak1, Bax, CytC, and Caspase3 mRNA and protein expression, inhibited Bcl2 mRNA and protein expression, and induced cell apoptosis in a dose effect. The Cu-caused changes of all above factors were alleviated by treatment with NAC. These results suggested that excessive Cu could induce oxidative stress and apoptosis via mitochondrial pathway in chicken hepatocytes.

Effect of binary surfactant mixtures on hydrophobic modification and moisture re-adsorption inhibition of lignite are investigated, along with that of dodecyltrimethylammonium bromide (DTAB), sodium dodecyl sulfonate (SDS) and dodecyl nonaethoxyl ether (C12(EO)9). Decrease in hydrophilicity of lignite is measured by determination of wetting heat and moisture re-adsorption rate of lignite before and after adsorbing surfactant. The differences in hydrophilicity change of lignite by using single and binary surfactant mixtures are discussed to disclose the synergistic or antagonism effect on hydrophobic modification of lignite. The experimental results show that, with adsorption of both single and binary mixtures of surfactants, lignite surface become hydrophobic. Compared to single component, DTAB-C12(EO)9 and DTAB-SDS display a higher efficiency on the hydrophilicity reduction and moisture re-absorption inhibition of lignite, while C12(EO)9-SDS does not. The calculated results indicate that there exists intermolecular synergism in the mixtures of DTAB-C12(EO)9 and DTAB-SDS, with forming synergetic aggregates “co-surfactant” DTAB-C12(EO)9 and DTAB-SDS, and C12(EO)9-SDS acts no synergism. These experimental results correspond to the calculation results of the properties of surfactant solution. Therefore, the solution properties of binary surfactant mixtures may provide a theoretical guiding in binary surfactant selection for reducing the hydrophilicity of lignite.

Cu is a metallic element that widely spread over in the environment, which have raised wide concerns about the potential toxic effects and public health threat. The objective of this study aimed to investigate the impression of copper (Cu)-triggered toxicity on mitochondrial dynamic, oxidative stress, and unfolded protein response (UPRmt) in fundic gland of pigs. Weaned pigs were randomly distributed into three groups, fed with different Cu of 10 mg/kg (control group), 125 mg/kg (group I), and 250 mg/kg (group Ⅱ). The trial persisted for 80 days and the fundic gland tissues were collected for further researches. Moreover, the markers participated to mitochondrial dynamic, UPRmt，and oxidative stress in fundic gland were determined. Results revealed that vacuolar degeneration were observed in the treated groups contrast with control group, and the Cu level was boosted with the increasing intake of Cu. Besides that, the levels of CAT, TRX, H2O2, and G6PDH were reduced in group Ⅰ and group Ⅱ, the mRNA levels of NRF2, HO-1, SOD-1, CAT, SOD-2, GSR, GPX1, GPX4, and TRX in the treated groups were promoted contrast to control group. Furthermore, the protein expression of KEAP1 was dramatically decreased, and the protein expression of NRF2, TRX and HO-1 were markedly enhanced in group Ⅰ and Ⅱ at 80 days. Moreover, the mRNA and protein expression levels of MFN1, MFN2, and OPA1 down-regulated and protein level of DRP1 was increased with the adding levels of Cu. Nevertheless, the UPRmt-related mRNA levels of CLPP, HTRA-2, CHOP, HSP10, and HSP60 were enhanced dramatically in Cu treatment group compared with control group. In general, our current study demonstrated that excessive absorption of Cu in fundic gland were related with stimulating UPRmt, oxidative stress, and the NRF2 interceded antioxidant defense. These results could afford an updated evidence on molecular theory of Cu-invited toxicity.

Despite the fact that copper (Cu) is a vital micronutrient to maintain body function, high doses of Cu through environmental exposure damage various organs, especially the liver, which is the main metabolic organ. To investigate the influence of long-term Cu-induced toxicity on mitophagy and apoptosis in rat liver, 96 seven-month-old male Sprague-Dawley rats were fed TBCC for 24 weeks. The results revealed that exposure to high Cu concentrations could promote oxidative stress liver injury by increasing the hepatic function index (ALT, AST and ALP) and MDA content, while reducing the activity of antioxidant enzymes (T-SOD, GSH-Px and CAT) related to oxidative stress. Consistent with histopathological observations, proper dietary Cu (15–60 mg/kg) could improve antioxidant stress levels and induce a dose-dependent increase in the mRNA expression of mitophagy-related genes, whereas a high Cu concentration (120 mg/kg) could cause severe liver impairment and ultrastructural changes and a reduction in mitophagosomes, accompanied by downregulation of Atg5, Beclin1, Pink1, Parkin, NIX, P62 and LC3B. The expression of apoptosis-related genes (Bax, Bax/Bcl-2, Caspase3, Cytc and p53) and proteins (Caspase3 and p53) was upregulated with the addition of dietary Cu. The results demonstrated that an appropriate dose of TBCC could improve liver function by promoting mitophagy and Cu enzymes that play antioxidative roles, while the accumulation of excess Cu could induce liver lesions by enhancing apoptosis and inhibiting mitophagy pathways.

Background: Copper (Cu), a common feed additive in diets for animals, is effective in improving growth performance and feed efficiency. However, excessive intake of Cu can cause toxic effects. Kidney is the main target organ of Cu, but the relationship between Cu-induced nephrotoxicity and its metabolic process remains unclear.Results: For deeply investigating the nephrotoxicity induced by Cu, a total of 240 broiler chicks were fed with different contents of Cu (11, 110, 220, and 330 mg/kg Cu) for 49 d in this study (60 chicks per group). The results of TUNEL staining showed that Cu could induce apoptosis in kidney with increasing of TUNEL-positive cells. Additionally, a total of 62 differential metabolites were detected by liquid chromatography-mass spectrometry (LC-MS), and mainly enriched in the metabolic pathways including riboflavin metabolism, glutathione metabolism, sphingolipid metabolism, and glycerophospholipid metabolism, which were closely related to mitochondrial metabolism. Meanwhile, the decrease of mitochondrial membrane potential, impairment of mitochondrial respiratory function, and the increase of mitochondrial membrane permeability indicated that renal mitochondria were damaged by excess Cu. Furthermore, the increase of mRNA and protein levels of Drp1, Bax, Bak-1, CytC, Caspase-3/cleaved Caspase-3 and the decrease of mRNA and protein levels of OPA1, Mfn1, Mfn2, and Bcl-2 confirmed that Cu could induce mitochondria-mediated apoptosis in kidney.Conclusions: These results highlighted that mitochondrial metabolism could be considered as an important factor in influencing Cu toxicity, which for further demonstrating that Cu could induce mitochondria-mediated apoptosis in kidney of broilers.

Receptor-interacting serine/threonine-protein kinase 3 (RIPK3) normally signals to necroptosis by phosphorylating MLKL. We report here that when the cellular RIPK3 chaperone Hsp90/CDC37 level is low, RIPK3 also signals to apoptosis. The apoptotic function of RIPK3 requires phosphorylation of the serine 165/threonine 166 sites on its kinase activation loop, resulting in inactivation of RIPK3 kinase activity while gaining the ability to recruit RIPK1, FADD, and caspase-8 to form a cytosolic caspase-activating complex, thereby triggering apoptosis. We found that PGF2α induces RIPK3 expression in luteal granulosa cells in the ovary to cause luteal regression through this RIPK3-mediated apoptosis pathway. Mice carrying homozygous phosphorylation-resistant RIPK3 S165A/T166A knockin mutations failed to respond to PGF2α but retained pro-necroptotic function, whereas mice with phospho-mimicking S165D/T166E homozygous knockin mutation underwent spontaneous apoptosis in multiple RIPK3-expressing tissues and died shortly after birth. Thus, RIPK3 signals to either necroptosis or apoptosis depending on its serine 165/threonine 166 phosphorylation status.

Fluorine (F) and its compounds produced from industrial production and coal combustion can cause air, water and soil contamination, which can accumulate in animals, plants and humans via food chain threatening public health. Fluoride exposure affects liver, kidney, gastrointestinal and reproductive system in humans and animals. Literature regarding fluoride influence on intestinal structure and microbiota composition in ducks is scarce. This study was designed to investigate these effects by using simple and electron microscopy and 16S rRNA sequencing techniques. Results indicated an impaired structure with reduced relative distribution of goblet cells in the fluoride exposed group. Moreover, the gut microbiota showed a significant decrease in alpha diversity. Proteobacteria, Firmicutes and Bacteroidetes were the most abundant phyla in both control and fluoride-exposed groups. Specifically, fluoride exposure resulted in a significant decrease in the relative abundance of 9 bacterial phyla and 15 bacterial genera. Among them, 4 phyla (Latescibacteria, Dependentiae, Zixibacteria and Fibrobacteres) and 4 genera (Thauera, Hydrogenophaga, Reyranella and Arenimonas) weren't even detectable in the gut microbiota of the ducks. In summary, higher fluoride exposure can significantly damage the intestinal structure and gut microbial composition in ducks.

Background The yield of microalgae biomass is the key to affect the accumulation of fatty acids. A few microalgae can assimilate organic carbon to improve biomass yield. In mixotrophic cultivation, microalgae can use organic carbon source and light energy simultaneously. The preference of the main energy source by microalgae determines the biomass yield. Auxenochlorella protothecoides is an oleaginous mixotrophic microalga that can efficiently assimilate glucose and accumulate a large amount of biomass and fatty acids. The current study focused on the effect of light on the growth and glucose assimilation of A. protothecoides. Results In this study, we found that the uptake and metabolism of glucose in A. protothecoides could be inhibited by light, resulting in a reduction of biomass growth and lipid accumulation. We employed comparative proteomics to study the influence of light on the regulation of glucose assimilation in A. protothecoides. Proteomics revealed that proteins involving in gene translation and photosynthesis system were up-regulated in the light, such as ribulose-phosphate 3-epimerase and phosphoribulokinase. Calvin cycle-related proteins were also up-regulated, suggesting that light may inhibit glucose metabolism by enhancing the production of glyceraldehyde-3-phosphate (G3P) in the Calvin cycle. In addition, the redox homeostasis-related proteins such as thioredoxin reductase were up-regulated in the light, indicating that light may regulate glucose uptake by changing the redox balance. Moreover, the increase of NADH levels and redox potential of the medium under illumination might inhibit the activity of the glucose transport system and subsequently reduce glucose uptake. Conclusions A theoretical model of how glucose assimilation in A. protothecoides is negatively influenced by light was proposed, which will facilitate further studies on the complex mechanisms underlying the transition from autotrophy to heterotrophy for improving biomass accumulation.

To explore the effects of copper (Cu) on energy metabolism and AMPK-mTOR pathway-mediated autophagy in kidney, a total of 240 one-day-old broiler chickens were randomized into four equal groups and fed on the diets with different levels of Cu (11, 110, 220, and 330 mg/kg) for 49 d. Results showed that excess Cu could induce vacuolar degeneration and increase the number of autophagosomes in kidney, and the adenosine triphosphate (ATP) level and mRNA levels of energy metabolism-related genes were decreased with the increasing dietary Cu level. Moreover, immunohistochemistry and immunofluorescence showed that the positive expressions of Beclin1 and LC3-II were mainly located in cytoplasm of renal tubular epithelial cells and increased significantly with the increasing levels of Cu. The mRNA levels of Beclin1, Atg5, LC3-I, LC3-II, Dynein and the protein levels of Beclin1, Atg5, LC3-II/LC3-I and p-AMPKα1/AMPKα1 were markedly elevated in treated groups compared with control group (11 mg/kg Cu). However, the mRNA and protein levels of p62 and p-mTOR/mTOR were significantly decreased with the increasing levels of Cu. These results suggest that impaired energy metabolism induced by Cu may lead to autophagy via AMPK-mTOR pathway in kidney of broiler chickens.

Copper (Cu) is a necessary trace mineral due to its biological activity. Excessive Cu can induce inflammatory response in humans and animals, but the underlying mechanism is still unknown. Here, 240 broilers were used to study the effects of excessive Cu on oxidative stress and NF-κB-mediated inflammatory responses in immune organs. Chickens were fed with diet containing different concentrations of Cu (11, 110, 220, and 330 mg of Cu/kg dry matter). The experiment lasted for 49 days. Spleen, thymus, and bursa of Fabricius (BF) on day 49 were collected for histopathological observation and assessment of oxidative stress status. Additionally, the mRNA and protein levels of NF-κB and inflammatory cytokines were also analyzed. The results indicated that excess Cu could increase the number and area of splenic corpuscle as well as the ratio of cortex and medulla in thymus and BF. Furthermore, excessive Cu intake could decrease activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px); but increase contents of malondialdehyde (MDA), TNF-α, IL-1, IL-1β; up-regulate mRNA levels of TNF-α, IFN-γ, IL-1, IL-1β, IL-2, iNOS, COX-2, NF-κB and protein levels of TNF-α, IFN-γ, NF-κB, p–NF–κB in immune organs. In conclusion, excessive Cu could cause pathologic changes and induce oxidative stress with triggered NF-κB pathway, and might further regulate the inflammatory response in immune organs of chicken.

Low plasma triiodothyronine (T3) concentration indicates nonthyroidal illness syndrome (NTIS), which might be associated with a poor outcome in patients in the intensive care unit (ICU). This study evaluated the relationship between NTIS and prognostic indicators in patients admitted to the ICU and examined the fT3 cut-off points that could be associated with 28-day mortality. This prospective observational study included patients admitted to the ICU of The Third Hospital of Hebei Medical University from February to November 2018. The baseline variables and the occurrence of low free T3 (FT3) were collected. The patients were divided into the NTIS (FT3<3.28) and non-NTIS groups. Among 305 patients, 118 (38.7%) were in the NTIS group. FT3 (P<0.001) and FT4 (P=0.001) were lower, while the 28-day mortality rate (P<0.001) and hospitalization expenses in ICU (P=0.001) were higher in the NTIS group. The univariable analyses identified NTIS, FT3, free thyroxine/FT3, APACHEII, sequential organ failure score, duration of mechanical ventilation, creatinine, oxygenation index, white blood cells, albumin, age, and brain natriuretic peptide as being associated with 28-day mortality (all P<0.05). The cut-off value of FT3 for 28-day mortality was 2.88 pmol/L. The 28-day mortality rate and hospitalization expenses in the ICU were higher in patients with NTIS. NTIS was independently associated with 28-day mortality.

Butachlor being an important member of chloroacetanilide herbicides, is frequently used in agriculture to control unwanted weeds. Exposure to butachlor can induce cancer, human lymphocyte aberration, and immunotoxic effects in animals. The current experimental trial was executed to determine the potential risks of herbicide butachlor to immunotoxicity and its mechanism of adverse effects on the spleen. For this purpose, mice were exposed to 8 mg/kg butachlor for 28 days, and the toxicity of butachlor on the spleen of mice was evaluated. We found that butachlor exposure led to an increase in serum ALB, GLU, TC, TG, and TP and changes in the morphological structure of the spleen of mice. More importantly, results showed that butachlor significantly increased the expression level of ATG-5, decreased the protein expression of LC3B and M-TOR, and significantly decreased the mRNA content of M-TOR and p62. Results revealed that the mRNA contents of APAF-1, CYTC, and CASP-9 related genes were significantly decreased after butachlor treatment. Subsequently, the mRNA levels of inflammatory cytokines (IL-1β, TNF-α, IL-10) were reduced in the spleen of treated mice. This study suggested that butachlor induce spleen toxicity and activate the immune response of spleen tissue by targeting the CYTC/BCL2/M-TOR pathway and caspase cascading activation of spleen autophagy and apoptosis pathways which may ultimately lead to immune system disorders.

Copper (Cu), a hazardous heavy metal, can lead to toxic effects on host physiology. Recently, specific mitochondria-localized miRNAs (mitomiRs) were shown to modulate mitochondrial function, but the underlying mechanisms remain undefined. Here, we identified mitomiR-1285 as an important molecule regulating mitochondrial dysfunction and mitophagy in jejunal epithelial cells under Cu exposure. Mitochondrial dysfunction and mitophagy were the important mechanisms of Cu-induced pathological damage in jejunal epithelial cells, which were accompanied by significant increase of mitomiR-1285 in vivo and in vitro. Knockdown of mitomiR-1285 significantly attenuated Cu-induced mitochondrial respiratory dysfunction, ATP deficiency, mitochondrial membrane potential reduction, mitochondrial reactive oxygen species accumulation, and mitophagy. Subsequently, bioinformatics analysis and luciferase reporter assay demonstrated that IDH2 was a direct target of mitomiR-1285. RNA interference of IDH2 dramatically reversed the effect that mitomiR-1285 knockdown relieved mitochondrial dysfunction and mitophagy induced by Cu, and the opposite effect was shown by overexpression of IDH2. Therefore, our results suggested that mitomiR-1285 aggravated Cu-induced mitochondrial dysfunction and mitophagy via suppressing IDH2 expression. These findings identified the important mechanistic connection between mitomiRs and mitochondrial metabolism under Cu exposure, providing a new insight into Cu toxicology.

Abundant oxygen-containing functional groups make lignite high moisture content, which reduces the utilization efficiency of lignite. The decrease in hydrophilicity of lignite surface can be achieved by treating with surfactant. In the present work, two kinds of nonionic surfactants with different hydrophilic groups, n-dodecyl β-D-maltoside (C12G2) and dodecyl hepta glycol (C12E7), were selected to modify the wettability of lignite surface by molecular dynamics simulation. Because of the drastic differences in compositions and structure of their headgroups, different behaviors were observed. The adsorption results of simulation indicate that polyhydroxy surfactant, C12G2, adsorbs strongly on lignite surface as a comparison to the poly ether surfactant, C12E7. However, the extent of hydrophobicity of modified lignite surface by these surfactants is inconsistent with their adsorption capabilities. Compared to the raw lignite, the hydrophilicity of lignite significantly decreases by adsorption of C12E7, while the C12G2 makes the lignite even more hydrophilicity. The strong polar oxygen-containing functional groups of lignite surface are covered by ethers in C12E7 with weaker polarity, which weakens the interaction between water and lignite. The hydrophilicity of lignite adsorbed C12G2 is strengthened due to the increase in surface polarity by the introduction of polar hydroxyl groups. The results of simulations are in accord with the available experimental data.

Decrease in hydrophilicity of lignite and moisture readsorption after adsorption of a series of lauryl polyoxyethylene ether (C12(EO)n) were studied with the measurement of wetting heat and moisture readsorption ratio. Effect of the hydrophilic lipophilic balance (HLB) of C12(EO)n on efficiency of the decrease in lignite hydrophilicity was investigated. X-ray photoelectron spectroscopy (XPS) and nitrogen adsorption/desorption were used to disclose the coverage characteristics of the different functional groups and the aggregations in pores of lignite by the adsorbed surfactant. Results indicated that both wetting heat and moisture readsorption ratio of lignite decreased significantly using different C12(EO)n. The size order of wetting heat of lignite after adsorption of different C12(EO)n with variation of the HLB values were almost consistent with that of moisture readsorption. The low HLB values of C12(EO)n especially with HLB of around 13.62 and 12.49/9.75 are most efficient respectively for the decrease in wetting heat and moisture readsorption of lignite. Coverage of oxygen-containing groups by surfactant was mainly devoted to the decrease in hydrophilicity of lignite. Among those oxygen groups in lignite surface, the COO/COOH and C-O/OH were responsible for adsorption priorities. Aggregations of surfactant in pores were beneficial for decreasing moisture readsorpiton of lignite surface due to the decreased storing space of moisture.

The Upper Paleozoic in the north part of Tianhuan depression in the Ordos Basin, NW China has lower hydrocarbon generation intensity and complex gas-water relationship, the main factors controlling the formation of tight sandstone gas and the distribution of tight sandstone gas in the low hydrocarbon generation intensity area are studied. Through two-dimensional physical simulation experiment of hydrocarbon accumulation, analysis of reservoir micro-pore-throat hydrocarbon system and dissection of typical gas reservoirs, the evaluation models of gas injection pressure, reservoir physical property, and gas generation threshold were established to determine the features of tight gas reservoirs in low hydrocarbon intensity area: (1) at the burial depth of less than 3 000 m, the hydrocarbon generation intensity of (7−10)×108 m3/km2 is high enough to maintain effective charging; (2) tight sandstone in large scale occurrence is conducive to accumulation of tight gas; (3) differences in reservoir physical property control the distribution of gas pool, for the channel sandstone reservoirs, ones with better physical properties generally concentrate in the middle of sandstone zone and local structural highs; ones with poor physical properties have low gas content generally. Based on the dissection of the gas reservoir in the north Tianhuan depression, the formation of tight gas reservoirs in low hydrocarbon generating intensity area are characterized by “long term continuous charging under hydrocarbon generation pressure, gas accumulation in large scale tight sandstone, pool control by difference in reservoir physical property, and local sweet spot”, and the tight gas pools are distributed in discontinuous “sheets” on the plane. This understanding has been proved by expanding exploration of tight sandstone gas in the north Tianhuan depression. Key words: Ordos Basin, Tianhuan depression, hydrocarbon generation intensity, tight sandstone gas, reservoir-forming mechanism, distribution pattern

Copper (Cu) is one of the ubiquitous environmental pollutants which have raised wide concerns about the potential toxic effects and public health threat. For deeply investigating the nephrotoxicity induced by Cu, the effects of Cu on mitochondria-mediated apoptosis in kidney were first to analyze by combining metabolomics and molecular biology techniques. In this study, broiler chicks were fed with different contents of Cu (11, 110, 220, and 330 mg/kg Cu) for 49 d. The results of terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) staining and transmission electron microscope showed that Cu could induce apoptosis in kidney, characterized by the increasing of TUNEL-positive cells and mitochondrial vacuolation. Additionally, a total of 62 differential metabolites were detected by liquid chromatography-mass spectrometry (LC-MS), and mainly enriched in the metabolic pathways including riboflavin metabolism, glutathione metabolism, sphingolipid metabolism, and glycerophospholipid metabolism, which were closely to mitochondrial metabolism. Meanwhile, the decreased mitochondrial membrane potential (MMP), increased mitochondrial membrane permeability and the change of mRNA and protein expression levels associated with mitochondria-mediated apoptosis and mitochondrial dynamics confirmed that Cu could induce mitochondria-mediated apoptosis. Therefore, our results demonstrated that Cu induced mitochondria-mediated apoptosis in kidney. Moreover, this study highlighted the metabolic characteristics of Cu to kidney, which suggested that mitochondrial metabolism could be considered as an important factor influencing toxicity.

The poor flotation behavior of low-rank coal is mainly caused by the high content of functional oxygen groups present on the coal surface. Polar collectors, especially oxygenated collectors, play a significant role in the flotation enhancement of low-rank coal. In this study, to improve the low-rank coal flotation performance, octanoic acid was selected as a polar collector and the corresponding promotion mechanism was investigated at the molecular scale theoretically. The flotation results showed that an increased low-rank coal flotation response can be obtained by using an octanoic acid collector compared to a nonpolar dodecane collector. To obtain a molecular-level explanation of this phenomenon, a method of density functional theory (DFT) combined with molecular dynamics simulation was employed to explore the involved micro-mechanism. DFT calculation results suggested that the octanoic acid collector can interact with benzoic acid and phenol (representatives of oxygen-containing groups on the low-rank coal surface) through hydrogen bonding and repel water molecules present on benzoic acid and phenol. Based on theoretical calculations, in the presence or absence of water molecules, hydrogen bonding and electrostatic interactions are responsible for the strong affinity of octanoic acid to carboxyl group and hydroxyl group on the low-rank coal surface. Therefore, it was favorable for octanoic acid to form a stable adsorption layer on the low-rank coal surface, thus contributing to low-rank coal flotation significantly.

The alternative development of coal-bearing hydrocarbon source rocks and low-porosity and low-permeability tight sandstone reservoirs of the Triassic Xujiahe Formation in the Sichuan Basin is favorable for near-source hydrocarbon accumulation. The natural gas composition of the Xujiahe Formation in the Sichuan Basin is dominated by hydrocarbon gases, of which the methane content is 80.16%–98.67%. Typically, the C2+ content is larger than 5% in main wet gas. The dry gas is mainly distributed in the western and northern regions of the basin. The non-hydrocarbon gases mainly contain nitrogen, carbon dioxide, hydrogen, and helium, with a total content of δ13C4 occurs in some natural gas with a low evolution degree; such gas is mainly coal-related gas from humic-type source rocks of the Xujiahe Formation. As for the natural gas, δ2HCH4 values ranged from −195‰ to −161‰, δ2HC2H6 values ranged from −154‰ to −120‰, and δ2HC3H8 values ranged from −151‰ to −108‰. The dry coefficient, δ13C and δ2HCH4 are all positively correlated with the maturity of source rocks. The higher the maturity of source rocks is, the larger the natural gas dry coefficient is and the larger the δ13C and δ2HCH4 values are, indicative of the characteristic of near-source accumulation. The δ2HC2H6 value of natural gas is influenced by paleosalinity to a relatively large extent; the higher the paleosalinity is, the larger the δ2HC2H6 value is. The Pr/Ph value of the condensate oil ranged from 1.60 to 3.43, illustrating light oxidization–light reduction and partial-oxidization characteristics of the depositional environment of coal-bearing source rocks of the Xujiahe Formation. The natural gas light hydrocarbon (C5–C7) from the Xujiahe Formation presented two characteristics: the first was the relatively high aromatic hydrocarbon content (19%–32.1%), which reveals the characteristic of natural gas with humic substances of high-maturity; the second was the low content of aromatic hydrocarbon (0.4%–9.3%), reflecting water-washing during the accumulation of the natural gas. The reported research outcomes indicate a potential mechanism for natural gas accumulation in the Xujiahe Formation, which will further guide natural gas exploration in this region.

Copper is an important trace mineral in the diet of poultry due to its biological activity. However, limited information is available concerning the effects of high copper on mitochondrial dysfunction. In this study, 72 broilers were used to investigate the effects of high dietary copper on liver mitochondrial dysfunction and electron transport chain defect. Birds were fed with different concentrations [11, 110, 220, and 330 mg of copper/kg dry matter (DM)] of copper from tribasic copper chloride (TBCC). The experiment lasted for 60 d. Liver tissues on d 60 were subjected to histopathological observation. Additionally, liver mitochondrial function was recorded on d 12, 36, and 60. Moreover, a site-specific defect in the electron transport chain in liver mitochondria was also identified by using various chemical inhibitors of mitochondrial respiration. The results showed different degrees of degeneration, mitochondrial swelling, and high-density electrons in hepatocytes. In addition, the respiratory control ratio (RCR) and oxidative phosphorylation rate (OPR) in liver mitochondria increased at first and then decreased in high-dose groups. Moreover, hydrogen peroxide (H2O2) generation velocity in treated groups was higher than that in control group, which were magnified by inhibiting electron transport at Complex IV. The results indicated that high dietary copper could decline liver mitochondrial function in broilers. The presence of a site-specific defect at Complex IV in liver mitochondria may be responsible for liver mitochondrial dysfunction caused by high dietary copper.

This clinical observation study aimed to investigate the relationship between the serum levels of vascular endothelial growth factor (VEGF) and its soluble receptors with the severity and the occurrence of late acute respiratory distress syndrome (ARDS) in early trauma.Sixty patients with multiple injuries were divided into three groups according to the Injury Severity Score (ISS) and the serum levels of VEGF, soluble VEGF receptor 1 (sVEGFR1), and sVEGFR2, were measured. Ten healthy people were recruited as controls. The incidence of late ARDS was also monitored, and its relationship to the above measures analyzed.VEGF was not associated with ISS (p0.05); sVEGFR1 was positively associated with ISS (r = 0.459, p0.0001); however, sVEGFR2 was negatively associated with ISS (r = 0.510, p0.0001). The serum VEGF levels between the ARDS group and the non-ARDS group showed no significant difference (p0.05). sVEGFR1 in the ARDS group was significantly higher than that in the non-ARDS group (p0.0001), and sVEGFR2 in the ARDS group was significantly lower than that in the non-ARDS group (p0.0001).In conclusion, the increasing of sVEGFR1 and the decreasing of sVEGFR2 in early trauma might be closely related to the occurrence of late ARDS.Prognostic study, level III.

To investigate the cellular mechanisms of multiple myeloma (MM), we used liquid chromatography-tandem mass spectrometry for proteomics analysis of CD138+ plasma cells from patients with MM and healthy controls. We found that the 60-kDa heat shock protein (HSP60, also known as HSPD1) was significantly upregulated in myeloma cells. HSP60 is an important chaperone protein that regulates the homeostasis of mitochondrial proteins and maintains mitochondrial function. Knockdown (KD) of HSP60 in myeloma cells resulted in inhibition of proliferation and reduced the quality of the mitochondria. Mitochondrial stress tests showed that HSP60 KD inhibited glycolysis and mitochondrial activity. Metabolomics showed a decrease in glycolysis and tricarboxylic acid cycle metabolites, and inhibited the formation of creatine and phosphocreatine by the reaction of S-adenosylmethionine (SAM) with amino acids mediated by demethyladenosine transferase 1, mitochondrial (TFB1M) and reduced energy storage substances. Moreover, HSP60 silencing influenced the synthesis of ribonucleotides and nicotinamide adenine dinucleotide phosphate (NADPH) by the pentose phosphate pathway to inhibit cell proliferation. HSP60 KD inhibited 5' adenosine monophosphate-activated protein kinase (AMPK), which inhibited the key enzyme 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3), effecting the metabolism of fatty acids by inhibiting malonyl-coenzyme A. Our data suggest that reduced HSP60 expression alters metabolic reprogramming in MM, inhibits tumour progression and reduces mitochondrial-dependent biosynthesis, suggesting that HSP60 is a potential therapeutic target for MM treatment.

Considering the predicted rising temperatures under current climate change and heat wave scenarios, organisms are expected to suffer more intense and frequent thermal stress. Induced heat is accumulated by organisms and can cause a variety of physiological stress responses. Ophraella communa is an effective biological control agent of common ragweed, Ambrosia artemisiifolia, but the responses of this biocontrol agent to heat stress have not been fully elucidated and, therefore, its potential responses to climate change are uncertain. We investigated the physiological metabolism of subsequent O. communa adults after: (1) different developmental stages (egg, larval, pupal, and adult) were exposed to thermal stress for 3 h each day for 3, 5, 5, and 5 days, respectively (by stage); and (2) individuals were exposed to thermal stress throughout the egg-to-adult period for 3 h each day. The high temperatures of 40, 42, and 44°C were used to induce thermal stress. A control group was reared at 28 ± 2°C. The results showed that short- or long-term exposure to daily phasic high temperatures significantly decreased water and lipid contents and significantly increased glycogen and glycerol contents in all adults (i.e., after exposure of different stages or throughout the egg-to-adult period). However, the total sugar content significantly increased in adults after the eggs and larvae were exposed to brief short-term thermal stress. Compared to the control, the total sugar content was also significantly higher in the adults and pupae exposed to 44°C. Total sugar content in females increased significantly in response to long-term phasic thermal stress at 40°C. However, sugar content of males exposed to 44°C decreased significantly. After long-term phasic thermal stress, water and glycogen contents in males were significantly higher than in females; however, females had higher total sugar and lipid contents. Therefore, our study provides a basic understanding of the metabolic responses of O. communa to thermal stress and offers insights into its potential as a natural biocontrol agent against A. artemisiifolia during the summer season and under predicted climate change scenarios.

Among different synthetic compounds copper (Cu) is persistently and frequently used as growth promoter, antibacterial, antifungal and antiparasitic agent and has become common environmental pollutant. Therefore, this study explores the cardio-toxic effects of control group (10 mg/kg bw Cu) and treatment group (125 and 250 mg/kg bw Cu), and it association with process of autophagy and metabolomics in myocardium of pigs kept in three different experimental treatments for a period of 80 days. The results of serum biochemical parameters showed a significantly increase in creatinine kinase (CK), creatine kinase-MB (CK-MB), high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C) and aspartate aminotransferase (AST) in pigs exposed to 125 mg/kg bw and 250 mg/kg bw Cu. Meanwhile, the severe structural abnormalities in cardiomyocytes were found when exposed to 250 mg/kg Cu at day 80. In addition, the mRNA and proteins (Beclin1, ATG5 and LC3II) expression levels were significantly increased and p62 was significantly decreased in cardiomyocytes exposed to 250 mg/kg Cu at day 80 of the trial. Further, UPLC-QTOF/MS technique showed that 7 metabolites were up-regulated and 37 metabolites were down-regulated in cardiomyocytes after 250 mg/kg Cu treatment, with a principal impact on the metabolic pathways including glycerophospholipid metabolism, one carbon pool by folate, fatty acid elongation and fatty acid degradation, which were related to autophagy. Overall, our study identified the autophagy processes and metabolites in metabolic pathways in Cu-induced myocardium injury, which provided useful evidence of myocardium toxicity caused by Cu exposure via metabolomics and multiple bioanalytic methods.

Numerous oxygen-containing groups present on the low rank coal (LRC) surface makes a poor flotation performance of LRC only using common oily collector. Increasing the spreading strength of oily collector on the LRC surface is of great significance for improving its flotation behavior. In this study, molecular dynamics simulation (MDS) was employed to investigate the microscopic spreading characteristics of non-polar oil droplet on the LRC surface. For this purpose, the configurations of non-polar oil droplet, spreading area, and interaction energy were calculated. It was found that on the raw LRC surface, the non-polar oil droplet remained spherical with a small spreading area and weak interaction strength. Besides, a non-ionic surfactant, dodecyl nonaethoxyl ether (C12EO9) was used to modify the surface wettability of LRC to enhance the non-polar oil droplet spreading. After surfactant pretreatment, a pancake-like oil film was formed on the LRC surface with a larger spreading area and more negative interaction energy compared with the case of the raw LRC surface. Based on these observations, the microscopic insights were provided about the spreading of non-polar oil droplet on the surface of LRC, as well as the role of surfactant enhancing its spreading intensity. In addition, the effect of the oxygen-containing groups on the spreading behaviors of non-polar oil droplet was also explored. The results suggested that carboxyl and hydroxyl are the main groups that hinder the spreading of the non-polar oil droplet on LRC surface. The simulation results are in accordance with the available experimental data.

The diet of broiler chickens supplemented with increasing concentrations of copper (Cu) was assessed for the effect of Cu on liver mitochondrial function. A total of 160, 1-d-old Cobb 500 broilers (Gallus domesticus) were randomly assigned in equal numbers into 4 groups, which differed in the concentration of copper supplements in the diet; 11 (control), 110, 220 and 330 mg of Cu/kg dry matter. Liver mitochondrial function was recorded at 12, 24, 36, 48 and 60 d of age. Supplementation with 110 mg Cu/kg dry matter enhanced mitochondrial function and activities of complexes I-V, and this was significant at 36 d of age compared with the other diets (P 0.05). Supplementation with 220 mg Cu/kg dry matter and 330 mg Cu/kg dry matter enhanced mitochondrial function and activities of complexes I-V at 12, 24 and 36 d of age, but displayed reduced function (P 0.05) at 48 and 60 d of age except in complex IV (P 0.05). Mitochondrial hydrogen peroxide (H2O2) production was also increased (P 0.05) with an increase of copper supplementation in the diet. The results indicate that appropriate dietary copper supplements are sufficient for improving mitochondrial function and activities of the respiratory complexes. Higher concentrations of copper, on the other hand, lead to copper toxicity by affecting certain respiratory complexes.

The aim of this study was to compare the oil content, fatty acid composition, and biodiesel properties of eight Prunus L. species in Inner Mongolia, to provide a basis for selecting appropriate species to produce biodiesel. In the eight species, oil content ranged from 38.92% to 58.82%. In Prunus pedunculata Pall., Prunus sibirica L., Prunus mongolica Maxim., Prunus humilis Bunge, Prunus ansu Kom., and Prunus triloba Lindl., oil contents were higher than 45%. Percentages of fatty acids varied greatly among different species, but all species contained a large percentage of critical oleic acid (35.03–77.02%). The biodiesel properties of seed kernel oil methyl ester of the eight species were determined and compared with the relevant specifications of ASTM D6751, EN 14214, and GB/T 20828. P. triloba Lindl., P. ansu Kom., P. pedunculata Pall., and P. sibirica L. were the most efficient feedstock for biodiesel production and suitable for further use and development.

Flotation collectors have a significant effect on the flotation performance and efficiency of fine low rank coal (LRC) particles. In this study, a mixed collector consisting of a nonpolar collector (dodecane) and a polar collector (C12EO4, tetraethylene glycol monododecyl ether) was used to improve the flotation performance of LRC. The results show that flotation behavior can be enhanced with the addition of C12EO4 in dodecane. The yield, combustible matter recovery, and flotation efficiency index increase with a larger volume ratio of C12EO4 in the mixed collector. A molecular dynamics simulation (MD) approach was applied to explore the microscopic strengthening mechanism of the mixed collector. The simulation demonstrated that the interaction strength and spreading ability of the mixed collector on the LRC surface increased due to electrostatic attraction and hydrogen bonding between the mixed collector and the LRC surface (as indicated by interaction energy and contact surface area), thereby enhancing flotation performance of the LRC. The effect of C12EO4 on the emulsification and dispersibility of dodecane is also discussed.

In this work, a combined molecular dynamics (MD) simulations and experiments were adopted to study the microscopic mechanism of the benzene ring in nonionic surfactants on low-rank coal (LRC) flotation. Two nonionic surfactants with identical headgroups, nonylphenol ethoxylate (NPEO) and dodecyl ethoxyl ethers (C12EO), were selected as the research objects. Two LRC surface models with varying metamorphism degrees were constructed. The rationality of the constructed surface models was firstly discussed from the chemical composition and structure. The morphology of the surfactants and the adsorption energy obtained by MD suggested that the arrangement of polar oxygen atoms in the surfactant depended on its interaction with the polar groups of LRC, and the presence of the benzene ring in the nonionic surfactant hardly changed its arrangement on the LRC surface. The benzene ring in the nonionic surfactant can enhance its adsorption on the higher rank coal surface via the π-bonds while it cannot promote its adsorption on the lower rank coal surface. The results obtained from the experiments are in accordance with the simulation results.

To study the effects of nonionic surfactants polarity on the quality of flotation clean low-rank coal (LRC), three nonionic surfactants of different polarity, n-dodecyl β-D-maltoside (DDM), nonylphenol ethoxylate (NPEO) and dodecyl ethoxyl ethers (C12EO) were selected and their flotation behaviors were experimentally investigated. By analysing the adsorption behaviors of surfactants and interfacial water on the kaolinite (0 0 1) surface and (0 0 1 ¯ ) surface, it could be concluded that the three surfactants could not replace the water molecules adsorbed on kaolinite (0 0 1) surface due to the strong adsorption between the water molecule and the kaolinite (0 0 1) surface. And the ability of the three surfactants to replace the water molecules adsorbed on kaolinite (0 0 1 ¯ ) surface is as follow: DDM > NPEO > C12EO, i.e. increasing the polarity of surfactants makes it easier to adsorb on kaolinite surface. In addition, the surfactants adsorption layer on kaolinite (0 0 1 ¯ ) surface is more compact with the increase in nonionic surfactant polarity. The results of simulations are in according with the experimental data. The presented study offers directions in surfactant selection and design in LRC cleaning.

The purpose of this research was to discuss the effects of copper (Cu)-induced toxicity on oxidative stress and autophagy in hypothalamus of broilers. In this study, 240 one-day-old broilers were randomly divided into 4 groups and the contents of dietary Cu in 4 groups were 11 mg/kg (control group), 110 mg/kg (group I), 220 mg/kg (group II), and 330 mg/kg (group III). The experiment lasted for 49 days and the hypothalamus tissues were collected for histological observation and detection of Cu content. Additionally, the indicators related to oxidative stress in hypothalamus were determined. Moreover, the mRNA expression levels of autophagy-related genes and the protein expression levels of Beclin1, LC3-II/LC3-I, and p62 in hypothalamus were measured. Results showed that the treated groups were observed vacuolar degeneration in hypothalamus compared to control group, and the Cu content in hypothalamus was increased with the increase of dietary Cu. Furthermore, the activities of SOD, CAT, T-AOC were increased in group I and group II and then decreased in group III, and the content of MDA and the mRNA levels of Nrf2, HO-1, SOD-1, CAT, GCLC, GCLM, and GST in treated groups were elevated compared to control group. Moreover, the mRNA expression levels of Beclin1, Atg5, LC3-I, LC3-II and the protein expression levels of Beclin1 and LC3-II/LC3-I up-regulated significantly with the increasing levels of Cu. However, the mRNA expression levels of p62 and mTOR and the protein expression level of p62 down-regulated remarkably. Taken together, our present study evidenced that excessive intake of Cu could induce oxidative stress and autophagy in hypothalamus of broilers.

The influence of water in samples on matrix effects and recoveries of analytes were investigated during the determination of residues of nine beta-agonists including salbutamol, terbutaline, cimaterol, fenoterol, clorprenaline, ractopamine, tulobuterol, clenbuterol, and penbuterol in porcine liver, muscle, and urine samples with liquid chromatography tandem mass spectrometry. Lyophilization procedure was adopted for sample moisture removal. The results showed that there were weak suppression effects for all target analytes except ractopamine, and no significant differences were found between fresh and lyophilized samples with respect to matrix effects of nine beta-agonists (P > 0.05). However, most analytes’ recoveries in the range of 85.0–115 % were obtained in lyophilized samples, while only a few ones were found in fresh samples. There are significant differences in recoveries for most analytes between fresh and lyophilized samples (P

Middle East respiratory syndrome coronavirus (MERS-CoV) infects host cells through binding the receptor binding domain (RBD) on its spike glycoprotein to human receptor dipeptidyl peptidase 4 (hDPP4). Here, we report identification of critical residues on hDPP4 for RBD binding and virus entry through analysis of a panel of hDPP4 mutants. Based on the RBD–hDPP4 crystal structure we reported, the mutated residues were located at the interface between RBD and hDPP4, which potentially changed the polarity, hydrophobic or hydrophilic properties of hDPP4, thereby interfering or disrupting their interaction with RBD. Using surface plasmon resonance (SPR) binding analysis and pseudovirus infection assay, we showed that several residues in hDPP4–RBD binding interface were important on hDPP4–RBD binding and viral entry. These results provide atomic insights into the features of interactions between hDPP4 and MERS-CoV RBD, and also provide potential explanation for cellular and species tropism of MERS-CoV infection., Highlights • It has been demonstrated that MERS-CoV infects host cells through binding its envelope spike (S) glycoprotein to the host cellular receptor dipeptidyl peptidase 4 (DPP4). • To identify the critical residues on hDPP4 for RBD binding and virus entry, we constructed a panel of hDPP4 mutants based on structure-guided mutagenesis. • Using surface plasmon resonance (SPR) binding analysis and pseudovirus infection assay, we showed that several residues on hDPP4 had significant impacts on virus/receptor interactions and viral entry. • Our study has provided new insights into the features of interactions between hDPP4 and MERS-CoV RBD, and provides potential explanation for cellular and species tropism of MERS-CoV infection.

Copper (Cu) is an essential trace element that is required for the catalysis of several cellular enzymes. Excessive Cu could induce hepatotoxicity in humans and multiple animals. The purpose of this study was to investigate the effects of autophagy machinery on Cu-induced hepatotoxicity. Chicken hepatocytes were cultured in medium in the absence and presence of Cu sulfate (CuSO4) (0, 10, 50, and 100 μM) for 0, 6, 12, and 24 h, and in the combination of CuSO4 and N-acetyl-l-cysteine (NAC) (1 mM), rapamycin (10 nM), and 3-methyladenine (3-MA) (5 mM) for 24 h. Results showed that Cu could markedly increase the number of autophagosomes and LC3 puncta, induce autophagy-related genes (Beclin1, ATG5, LC3Ⅰ, LC3Ⅱ, mTOR, and Dynein) mRNA expression and proteins (BECN1, LC3Ⅱ/LC3Ⅰ) expression. NAC could relieve Cu-induced the changes of above genes and proteins. Additionally, rapamycin attenuated Cu-induced the increased lactic dehydrogenase (LDH), aspartate amino transferase (AST), and alanine aminotransferase (ALT) activities, and SOD-1 mRNA expression as well as the decreased cell viability, reactive oxygen species (ROS), hydrogen peroxide, total superoxide dismutase (T-SOD), malonaldehyde (MDA), catalase (CAT), HO-1 mRNA expression, adenosine triphosphate (ATP) levels, mitochondrial mass, and mitochondria membrane potential (MMP). But 3-MA had the opposite effects on above factors. Collectively, these findings provide strong evidence that Cu could induce autophagy by generating excessive ROS in hepatocytes, and autophagy might attenuate Cu-induced mitochondrial dysfunction by regulating oxidative stress.

ρ-Cymene was one of the major components of volatiles released by croftonweed. The allelopthy of ρ-cymene on the growth of upland rice seedlings was performed. Hydrogen peroxide generation, malondialdehyde (MDA) content, proline content, total ascorbate (ascorbate/dehydroascorbate), reduced/oxidized glutathione, and the levels of induction of antioxidant enzyme were studied in the seedlings of upland rice. ρ-Cymene inhibited the growth of upland rice seedlings. Exposure of upland rice seedlings to ρ-cymene increased levels of H2O2, MDA, and proline, indicating lipid peroxidation and induction of oxidative stress. Activities of the antioxidant enzymes superoxide dismutase, catalase, peroxidase, guaiacol peroxidase, ascorbate peroxidase, and glutathione reductase were significantly elevated during the treatment period (7–15 d) compared with enzymes in the upland rice seedlings unexposed to ρ-cymene, thereby indicating the enhanced generation of reactive oxygen species (ROS) upon ρ-cymene exposure. These results suggest that activation of the antioxidant system by ρ-cymene led to the formation of ROS that resulted in cellular damage and decreased growth of upland rice seedlings.

Several volatile allelochemicals were identified and characterized from fresh leaf tissues of the invasive croftonweed. A simple bioassay was used to demonstrate the release of volatile allelochemicals from leaf tissues. The bioassays revealed that foliar volatile components of croftonweed exhibited significant effects on the seedling growth of upland rice. Peroxidase (POD) activity, superoxide dismutase (SOD) activity, and root oxidizability rose as the concentration of volatiles increased. Activity for both POD and SOD significantly increased with exposure to 15 g and 20 g of croftonweed leaf tissue for 5 d. Root activity was significant at 10 g compared to the control. The volatile components also stimulated the development of the aerenchyma tissue and inhibited lateral root formation. Leaf volatiles of croftonweed were identified by gas chromatography coupled with mass spectrometry (GC–MS). Some of the compounds identified included α-phellandrene, camphene, ρ-cymene, 2-carene, α-pinene, limonene, and (z)-3-hexen-1-ol. Bioassays showed that four of these compounds could account for the observed phytotoxicity imparted by total leaf volatiles. Limonene, 2-carene, α-pinene and camphene had no phytotoxic effect on shoot elongation. Phellandrene did cause inhibition in shoot growth at all concentrations. Both (z)-3-hexene-1-ol and ρ-cymene inhibited both shoot elongation and root elongation, but the effects of the two compounds on root length were more significant than on the shoot length.

Abies fabri (Mast.) Craib is an endemic and dominant species in typical sub-alpine dark coniferous forests distributed in mountainous regions of the eastern Tibetan Plateau, China. We investigated the ecophysiological responses of A. fabri seedlings to short-term artificially-applied drought, nitrogen addition alone, and the combination of these treatments. Drought was created by excluding natural precipitation with an automatically controlled plastic roof that covered the seedlings. Nitrogen fertilization was applied weekly by spraying over seedlings with ammonium nitrate solution. Experiment results showed that drought caused a reduction in photosynthetic nitrogen use efficiency and leaf mass per area. Nitrogen addition enhanced photosynthetic performance by increasing net photosynthetic rate. In the drought plots, nitrogen addition increased net photosynthetic rate and instantaneous water use efficiency. These results showed that applied nitrogen improved plant water use efficiency and N accumulation in plant organs under drought conditions. Especially under drought conditions more N was concentrated into needles by applied nitrogen as compared with other organs. In conclusion, our results indicated that the combination of nitrogen addition and drought may result in positive effects on A. fabri seedlings in the short-term.