Native promoter-mediated transcriptional regulation of crucial oleosin protein OLE1 from Prunus sibirica for seed development and high oil accumulation.

Oleosin (OLE) is vital to stabilize lipid droplet for seed triacylglycerol (TAG) storage. This work aimed to determine key OLE and to unravel mechanism that governed seed oil accumulation of Prunus sibirica for developing biodiesel. An integrated assay of global identification of LD-related protein and the cross-accessions/developing stages comparisons associated with oil accumulative amount and OLE transcript level was performed on seeds of 12 plus trees of P. sibirica to identify OLE1 (15.5 kDa) as key oleosin protein crucial for high seed oil accumulation. The OLE1 gene and its promoter were cloned from P. sibirica seeds, and overexpression of PsOLE1 in Arabidopsis was conducted under the controls of native promoter and constitutive CaMV35S promoter, respectively. PsOLE1 promoter had seed-specific cis-elements and showed seed specificity, by which PsOLE1 was specifically expressed in seeds. Ectopic overexpression of PsOLE1, especially driven by its promoter, could facilitate seed development and oil accumulation with an increase in unsaturated FAs, and upregulate transcript of TAG assembly enzymes, but suppress transcript of LD/TAG-hydrolyzed lipases and transporters, revealing a role of native promoter-mediated transcription of PsOLE1 in seed development and oil accumulation. PsOLE1 and its promoter have considerable potential for engineering oil accumulation in oilseed plants.

Dietary Macroalgae Saccharina japonica Ameliorates Liver Injury Induced by a High-Carbohydrate Diet in Swamp Eel (Monopterus albus).

A high-carbohydrate diet lowers the rearing cost and decreases the ammonia emission into the environment, whereas it can induce liver injury, which can reduce harvest yields and generate economic losses in reared fish species. Macroalgae Saccharina japonica (SJ) has been reported to improve anti-diabetic, but the protective mechanism of dietary SJ against liver injury in fish fed a high-carbohydrate diet has not been studied. Therefore, a 56-day nutritional trial was designed for swamp eel Monopterus albus, which was fed with the normal diet [20% carbohydrate, normal carbohydrate (NC)], a high carbohydrate diet (32% carbohydrate, HC), and a HC diet supplemented with 2.5% SJ (HC-S). The HC diet promoted growth and lowered feed coefficient (FC), whereas it increased hepatosomatic index (HSI) when compared with the NC diet in this study. However, SJ supplementation increased iodine contents in muscle, reduced HSI, and improved liver injury, such as the decrease of glucose (GLU), total bile acid (TBA), and alanine aminotransferase (ALT) in serum, and glycogen and TBA in the liver. Consistently, histological analysis showed that SJ reduced the area of lipid droplet, glycogen, and collagen fiber in the liver (p < 0.05). Thoroughly, the underlying protective mechanisms of SJ supplementation against HC-induced liver injury were studied by liver transcriptome sequencing coupled with pathway analysis. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of the differentially expressed genes (DEGs), such as the acetyl-coenzyme A synthetase (acss1), alcohol dehydrogenase (adh), interferon-induced protein with tetratricopeptide repeats 1 (ifit1), aldo-keto reductase family 1 member D1 (akr1d1), cholesterol 7-alpha-monooxygenase (cyp7a1), and UDP-glucuronosyltransferase (ugt), indicated that the pathway of glycolysis/gluconeogenesis was the main metabolic pathway altered in the HC group compared with the NC group. Meanwhile, hepatitis C, primary BA biosynthesis, and drug metabolism-cytochrome P450 were the three main metabolic pathways altered by SJ supplementation when compared with the HC group. Moreover, the BA-targeted metabolomic analysis of the serum BA found that SJ supplementation decreased the contents of taurohyocholic acid (THCA), taurochenodeoxycholic acid (TCDCA), taurolithocholic acid (TLCA), nordeoxycholic acid (NorDCA), and increased the contents of ursocholic acid (UCA), allocholic acid (ACA), and chenodeoxycholic acid (CDCA). In particular, the higher contents of UCA, ACA, and CDCA regulated by SJ were associated with lower liver injury. Overall, these results indicate that the 2.5% supplementation of SJ can be recommended as a functional feed additive for the alleviation of liver injury in swamp eel-fed high-carbohydrate diets.

Progress in traditional Chinese medicine and natural extracts for the treatment of lupus nephritis.

Lupus nephritis (LN) is an autoimmune disease with multiple system involvement and is also one of the most serious forms of organ damage in systemic lupus erythematosus (SLE), which is mainly caused by the formation and deposition of immune complexes in glomeruli. More than 50% of SLE patients have clinical manifestations of renal damage. At present, the treatment of lupus nephritis is mainly based on glucocorticoids and immunosuppressants. However, due to adverse drug reactions and frequent recurrence or aggravation after drug reduction or withdrawal, the prognosis remains poor; thus, it is still one of the most important causes of end-stage renal failure. Therefore, new treatment strategies are urgently needed. This article aims to review the application of traditional Chinese medicine and natural extracts in the treatment of lupus nephritis to provide the basic mechanisms of treatment and a new treatment strategy with clear effects and high safety performance.

Characterization of genes encoding ω-6 desaturase PoFAD2 and PoFAD6, and ω-3 desaturase PoFAD3 for ALA accumulation in developing seeds of oil crop Paeonia ostii var. lishizhenii.

Paeonia ostii var. lishizhenii has emerged as a valuable oil-producing crop with splendid characteristic of high α-linolenic acid (C18:3, ALA) content in its seed oil for healthy food supplement, but the molecular mechanism for seed ALA accumulation remains enigmatic. In our previous report, a PoSAD gene encoding stearoyl-ACP desaturase had been cloned and functional charactered for the first desaturation procedure involved in ALA biosynthesis pathway in P. ostii var. lishizhenii endosperms, while other participants have not been identified to date. In this study, full-length cDNAs of PoFAD2 (1489 bp), PoFAD6 (1638 bp), and PoFAD3 (1709 bp) were isolated based on our recent transcriptome sequencing data. Bioinformatic analyses revealed that the PoFADs were closest to their counterparts from Paeoniaceae species P. ludlowii, P. rockii, and P. suffruticosa in phylogenetic tree, which shared highly conserved histidine boxes (HXXXH, HXXHH, and HXXHH), exhibiting typical characters of membrane-bound desaturases in higher plants. Additionally, the PoFAD2 and PoFAD3 were specifically expressed and highly associated with LA and ALA accumulation in developing endosperms, whereas PoFAD6 expression has no significantly difference during whole seed developing stages. The catalytic function of these PoFADs were further analyzed by heterologous expression in Saccharomyces cerevisiae and Arabidopsis thaliana. The results showed that PoFAD2 and PoFAD6 could catalyze linoleic acid (C18:2) synthesis, while PoFAD3 had ability to produce ALA. This study functional identified three PoFAD genes, which indicates their critical roles in ALA biosynthesis pathway in P. ostii var. lishizhenii, and is of great theoretical and practical meaning on breeding and cultivating new tree peony varieties to promote human health and nutrition supplement.

Melatonin increases leaf disease resistance and saponin biosynthesis in Panax notogiseng.

Panax notoginseng (Bruk.) FH Chen is a valuable traditional herb in China, with saponins being the main medicinal components in its roots. However, leaf diseases are a major factor limiting growth and production of P. notoginseng. Melatonin is a ubiquitous signaling molecule associated with abiotic stress resistance. In this study, we investigated the role of melatonin in leaf disease resistance of P. notoginseng in field conditions. Additionally, saponin concentrations were analyzed to evaluate the suitability of melatonin use in agricultural practice. Our results showed that exogenous application of melatonin promoted the endogenous phytomelatonin accumulation via upregulation of genes involved in its biosynthesis. The application of 10 µM melatonin decreased the incidence of leaf diseases (gray mold, round spot, and black spot) by about 40% compared with the solvent control, which might have been due to the increased expression of genes associated with immunity and disease resistance. Furthermore, concentrations of saponins and expression of their biosynthesis-related genes were significantly increased by melatonin. Taken together, the data presented here suggested that melatonin could be used in agricultural management of P. notoginseng because it increased leaf disease resistance and biosynthesis of saponins.

Methyl-Donor Micronutrient for Gestating Sows: Effects on Gut Microbiota and Metabolome in Offspring Piglets.

This study aimed to investigate the effects of maternal methyl-donor micronutrient supplementation during gestation on gut microbiota and the fecal metabolic profile in offspring piglets. Forty-three Duroc × Erhualian gilts were assigned to two dietary groups during gestation: control diet (CON) and CON diet supplemented with MET (folic acid, methionine, choline, vitamin B6, and vitamin B12). The body weights of offspring piglets were recorded at birth and weaning. Besides this, fresh fecal samples of offspring piglets were collected at 7, 14, and 21 days. The gut microbiota composition, metabolic profile, and short-chain fatty acid (SCFA) profiles in the fecal samples were determined using 16S rDNA sequencing, liquid chromatography-mass spectrometry metabolomics, and gas chromatography methods, respectively. The results showed that maternal methyl-donor micronutrient supplementation increased the microbiota diversity and uniformity in feces of offspring piglets as indicated by increased Shannon and Simpson indices at 7 days, and greater Simpson, ACE, Chao1 and observed species indices at 21 days. Specifically, at the phylum level, the relative abundance of Firmicutes and the Firmicutes to Bacteroidetes ratio were elevated by maternal treatment. At the genus level, the relative abundance of SCFA-producing Dialister, Megasphaera, and Turicibacter, and lactate-producing Sharpea as well as Akkermansia, Weissella, and Pediococcus were increased in the MET group. The metabolic analyses show that maternal methyl-donor micronutrient addition increased the concentrations of individual and total SCFAs of 21-day piglets and increased metabolism mainly involving amino acids, pyrimidine, and purine biosynthesis. Collectively, maternal methyl-donor micronutrient addition altered gut microbiota and the fecal metabolic profile, resulting in an improved weaning weight of offspring piglets.

Spatial patterns and influencing factors of sewage treatment plants in the Guangdong-Hong Kong-Macau Greater Bay Area, China.

Over the past few years the discharge of waste and sewage in the Guangdong-Hong Kong-Macau Greater Bay Area (GHMB) of China has increased, exerting a great amount of pressure on ecological protection. In this study, we focus on achieving a balanced spatial layout of sewage treatment plants in order to reveal the regional differences and spatial patterns of sewage treatment plants in the GHMB and identify the key factors influencing the spatial patterns. In particular, we employ POI (point of interest) geographical data to evaluate the spatial patterns and agglomeration status of sewage treatment plants in the GHMB using Exploratory Spatial Data Analysis (ESDA). We then explore the principle influencing factors of the determined spatial patterns using the geographical weighted regression model (GWR). Results demonstrate that: (1) the absolute number of sewage treatment plants is highest in the central and western regions, while the per capita of sewage treatment plants is clearly clustered in the northwest and southwest regions; (2) the absolute number of sewage treatment plants exhibits larger spatial dissimilarity than that of the per capita values, with High-High cluster types principally distributed in the conjunction areas of Jiangmen, Foshan and Zhaoqing in western GHMB and Low-Low clusters in the western Pearl River Delta (PRD) estuary; and (3) the key influencing factors are identified as GDP per capita, the output value of the primary and secondary industries and industrial water consumption. Regions with high regression coefficient fluctuations of the four influencing factors are centralized within the PRD estuary and adjacent regions. Policy recommendations including optimizing the sewage treatment plant layout, enhancing the service capacity potential of the existing sewage treatment plants and upgrading the industrial structure are proposed to promote a spatial equilibrium configuration of sewage treatment plants in the GHMB.

Maternal Methyl-Donor Micronutrient Supplementation During Pregnancy Promotes Skeletal Muscle Differentiation and Maturity in Newborn and Weaning Pigs.

Adequate maternal methyl-donor micronutrient (MET) intake is an important determinant of the organ development and metabolic renovation of offspring. The mechanism involved in skeletal myogenesis and the effect of MET supplementation during pregnancy on the maternal body remain unclear. Thus, this study aimed to investigate the potential effect of methyl donor micronutrients (MET) on skeletal muscle development and metabolism in offspring using pig models. Forty-three Duroc × Erhualian gilts were assigned to two dietary groups during gestation: control diet (CON) and CON diet supplemented with MET (folic acid, methionine, choline, vitamin B6, and vitamin B12). The results showed that maternal MET exposure during pregnancy significantly increased the concentrations of protein, triiodothyronine (T3), and thyroxine (T4) in colostrum and methyl metabolites, including S-adenosylmethionine (SAM), S-adenosyl-L-homocysteine (SAH), 5-methyl-tetrahydrofolate (5-MTHF), and betaine, in the maternal and offspring umbilical vein serum. A similar pattern was demonstrated in the body weight gain and myofiber diameters in offspring. In addition, maternal MET supplementation significantly increased the concentration of offspring serum insulin-like growth factor 1 (IGF-1), T3, and T4; upregulated the mRNA expression of IGF-1 and IGF-1 receptor (IGF-1r) and the phosphorylation level of protein kinases in offspring longissimus dorsi muscle; and upregulated the expression of myogenic genes and fast myosin heavy chain (fast MyHC) in offspring skeletal muscle. Supplementing sows with higher levels of MET during gestation may promote skeletal muscle differentiation and maturity and improve the skeletal muscle mass of the piglets.

Dietary guanidinoacetic acid improves the growth performance and skeletal muscle development of finishing pigs through changing myogenic gene expression and myofibre characteristics.

This study aimed to evaluate the effects of dietary guanidine acetic acid (GAA) supplementation on growth performance, carcass traits and the expression of muscle growth-related genes in finishing pigs. A total of 128 (81.03 ± 1.09 kg body weight) crossbred pigs (Duroc × Landrace ×Yorkshire) were blocked by body weight and allotted to 16 pens (eight pigs per pen), and pens were randomly assigned within blocks to one of five dietary treatments, with a basal diet (control group) or a basal diet supplemented with 0.03%, 0.06% and 0.09% GAA respectively. During the 60-day trial, GAA increased the average dairy gain (ADG) and average daily feed intake (ADFI) (p < .05). The back fat thickness of pigs fed 0.06% GAA was lower than other groups (p < .05). Pigs fed 0.06% GAA had improved lean meat percentage, loin muscle area, shear force and cross-sectional area of muscle fibre in comparison with control group (p < .05). The drop loss and the muscle fibre density in pigs fed 0.06% GAA were lower than control (p < .05). In addition, dietary GAA enhanced the expression of myosin heavy chain gene (MYH4), myogenic determination (Myod) and myogenic factor 5 (Myf5) in longissimus dorsi and carnitine palmitoyltransferase-1(CPT-1) in liver (p < .05). Meanwhile, GAA decreased the expression of Myostatin in longissimus dorsi and fatty acid synthase (FAS) in liver (p < .05). In conclusion, our results showed that appropriate dietary GAA supplementation (0.06%) promotes skeletal muscle development through changing myogenic gene expression and myofibre characteristics.

Microfabrication-Based Three-Dimensional (3-D) Extracellular Matrix Microenvironments for Cancer and Other Diseases.

Exploring the complicated development of tumors and metastases needs a deep understanding of the physical and biological interactions between cancer cells and their surrounding microenvironments. One of the major challenges is the ability to mimic the complex 3-D tissue microenvironment that particularly influences cell proliferation, migration, invasion, and apoptosis in relation to the extracellular matrix (ECM). Traditional cell culture is unable to create 3-D cell scaffolds resembling tissue complexity and functions, and, in the past, many efforts were made to realize the goal of obtaining cell clusters in hydrogels. However, the available methods still lack a precise control of cell external microenvironments. Recently, the rapid development of microfabrication techniques, such as 3-D printing, microfluidics, and photochemistry, has offered great advantages in reconstructing 3-D controllable cancer cell microenvironments in vitro. Consequently, various biofunctionalized hydrogels have become the ideal candidates to help the researchers acquire some new insights into various diseases. Our review will discuss some important studies and the latest progress regarding the above approaches for the production of 3-D ECM structures for cancer and other diseases. Especially, we will focus on new discoveries regarding the impact of the ECM on different aspects of cancer metastasis, e.g., collective invasion, enhanced intravasation by stress and aligned collagen fibers, angiogenesis regulation, as well as on drug screening.