In vitro generation of genetic diversity for directed evolution by error-prone artificial DNA synthesis.

Generating genetic diversity lies at the heart of directed evolution which has been widely used to engineer genetic parts and gene circuits in synthetic biology. With the ever-expanding application of directed evolution, different approaches of generating genetic diversity are required to enrich the traditional toolbox. Here we show in vitro generation of genetic diversity for directed evolution by error-prone artificial DNA synthesis (epADS). This approach comprises a three-step process which incorporates base errors randomly generated during chemical synthesis of oligonucleotides under specific conditions into the target DNA. Through this method, 200 ~ 4000 folds of diversification in fluorescent strength have been achieved in genes encoding fluorescent proteins. EpADS has also been successfully used to diversify regulatory genetic parts, synthetic gene circuits and even increase microbial tolerance to carbenicillin in a short time period. EpADS would be an alternative tool for directed evolution which may have useful applications in synthetic biology.

Exosomal MALAT1 from macrophages treated with high levels of glucose upregulates LC3B expression via miR-204-5p downregulation.

BACKGROUND: Metastasis-associated lung adenocarcinoma transcript 1 ( MALAT1 ) plays a critical role in the pathophysiology of diabetes-related complications. However, whether macrophage-derived MALAT1 affects autophagic activity under hyperglycemic conditions is unclear. Therefore, we investigated the molecular regulatory mechanisms of macrophage-derived MALAT1 and autophagy under hyperglycemic conditions. METHODS: Hyperglycemia was induced by culturing macrophages in 25 mM glucose for 1 hour. Exosomes were extracted from the culture media. A rat model of carotid artery balloon injury was established to assess the effect of MALAT1 on vascular injury. Reverse transcription, real-time quantitative polymerase chain reaction, western blotting, immunohistochemical staining, and luciferase activity assays were performed. RESULTS: Stimulation with high levels of glucose significantly enhanced MALAT1 expression in macrophage-derived exosomes. MALAT1 inhibited miR-204-5p expression in macrophage-derived exosomes under hyperglycemic conditions. siRNA-induced silencing of MALAT1 significantly reversed macrophage-derived exosome-induced miR-204-5p expression. Hyperglycemic treatment caused a significant, exosome-induced increase in the expression of the autophagy marker LC3B in macrophages. Silencing MALAT1 and overexpression of miR-204-5p significantly decreased LC3B expression induced by macrophage-derived exosomes. Overexpression of miR-204-5p significantly reduced LC3B luciferase activity induced by macrophage-derived exosomes. Balloon injury to the carotid artery in rats significantly enhanced MALAT1 and LC3B expression, and significantly reduced miR-204-5p expression in carotid artery tissue. Silencing MALAT1 significantly reversed miR-204-5p expression in carotid artery tissue after balloon injury. MALAT1 silencing or miR-204-5p overexpression significantly reduced LC3B expression after balloon injury. CONCLUSION: This study demonstrated that hyperglycemia upregulates MALAT1 . MALAT1 suppresses miR-204-5p expression and counteracts the inhibitory effect of miR-204-5p on LC3B expression in macrophages to promote vascular disease.

Effect of DBD Plasma Treatment on Activity of Mo-Based Sulfur-Resistant Methanation Catalyst.

By combining the advantages of dielectric barrier discharge (DBD) low temperature plasma and fluidized bed, the effect of plasma on the performance of supported Mo-based catalyst was studied in this paper. The performance of the catalyst obtained by plasma treatment, calcined, plasma+calcined was compared, and the appropriate catalyst preparation scheme was explored. Comparing with the three catalysts, it was concluded that the catalyst average conversion after 30 W plasma treatment is 33.40 %, which was 8.94 % and 12.75 % higher than the other two, respectively. The structure and properties of the catalyst were characterized by N2-Physisorption, H2-chemisorption, XRD, TEM, XPS, Raman and NO-pulse adsorption. Then, by analyzing the characterization results, it can be seen that plasma can make the catalyst have a higher specific surface area and a more dispersed active metal with smaller grain size. Through the surface species identification characterization, it was found that plasma can produce more defective structures and expose more active sites, which is the main reason for the difference in conversion.

Impact of Functional Conservation Surgery on Sleep Respiration of Patients with Glottic Cancer.

Objective: Laryngeal cancer is a common tumor in the head and neck, and surgery is one of the main treatment methods for laryngeal cancer. Laryngeal cancer surgery destroys the laryngeal cartilage scaffold, leading to structural changes in the laryngeal cavity and affecting respiratory compliance during sleep. However, less attention has been paid to the impact of changes in laryngeal structure on sleep breathing conditions. This article conducts a prospective study on the effects of preserving laryngeal function in cancer glottic surgery on sleep and respiratory status in patients, in order to understand the preoperative and postoperative OSAHS(obstructive sleep apnea-hypopnea syndrome) of glottic cancer patients , as well as the impact of surgery on OSAHS of patients. Provide a reference for improving the sleep quality of postoperative patients with laryngeal cancer. Methods: 47 patients with glottic cancer who underwent laryngeal function preservation surgeries were studied. They are divided into 28 cases of T1 and T2, who underwent vertical lateral frontal partial laryngectomy (VLFPL), and 19 cases of T3 and T4 who underwent vertical lateral frontal subtotal laryngectomy (VLFSL). All patients' sleep breathing statuses were recorded using a portable sleep breathing monitor one week before surgery and two months after the removal of the tracheal cannula. Analyze the proportion of OSAHS in the 47 patients before and after surgery and compare the obstructive sleep apnea scores of these patients using the Wilcoxon rank sum test of paired grade data. The paired data t-test was used to analyze the apnea-hypopnea index (AHI), apnea index (AI), hypopnea index (HI), minimum blood oxygen saturation (LSaO2), and mean blood oxygen saturation (MSaO2) of all study subjects, patients undergoing VLFPL and VLFSL. Results: 1. Among the 47 patients with glottic cancer, 42.6% (20 / 47) were in line with OSAHS before the operation, and 57.4% (27 / 47) were in line with OSAHS after the operation. 47 patients showed an increasing trend in the OSAHS scores (no, mild, moderate, and severe) after surgery compared to the before-surgery scores, and the difference was statistically significant (P < 0.05). 2. Among the 47 study subjects, AHI and HI increased after surgery compared to preoperative, while LSaO2 and MSaO2 decreased after surgery compared to preoperative (P < .05); There was no statistically significant difference between postoperative and preoperative AI (P > .05). Among 28 patients undergoing VLFPL, LSaO2 decreased after surgery compared to before surgery (P < .05) and there was no statistically significant difference in AHI, AI, MSaO2, and HI after surgery compared to before surgery (P > .05). Among 19 patients undergoing VLFSL, AHI, and HI increased after surgery, while LSaO2 and MSaO2 decreased after surgery (P < .05) and there was no statistically significant difference in postoperative AI compared to preoperative (P > .05). Conclusion: The prevalence of OSAHS in patients with glottic cancer before and after surgery was higher than that in the general population. The effect of functional preservation surgery on sleep breathing in patients with glottic cancer is related to the degree of destruction of the thyroid cartilage scaffold and the scope of surgery. Surgery mainly increases the degree of OSAHS by aggravating patients' hypoventilation rather than apnea.

A New Sufficient & Necessary Condition for Testing Linear Separability Between Two Sets.

As a fundamental mathematical problem in the field of machine learning, the linear separability test still lacks a theoretically complete and computationally efficient method. This paper proposes and proves a sufficient and necessary condition for linear separability test based on a sphere model. The advantage of this test method is two-fold: (1) it provides not only a qualitative test of linear separability but also a quantitative analysis of the separability of linear separable instances; (2) it has low time cost and is more efficient than existing test methods. The proposed method is validated through a large number of experiments on benchmark datasets and artificial datasets, demonstrating both its correctness and efficiency.

Effects of dietary vitamin K3 supplementation on production performance, egg quality, vitamin K-dependent proteins, and antioxidant properties in breeding geese during the laying period.

The aim of this work was to investigate the effects of dietary vitamin K3 (VK3) supplementation on production performance, egg quality, vitamin K-dependent proteins, and antioxidant properties in breeding geese during the laying period. A total of one hundred twenty 82-wk-old Wulong geese with similar body weights were randomly divided into 6 groups with 4 replicates and 5 geese each (1 male and 4 female). The geese in the control group were fed a basal diet, and the geese in the treatment groups were fed diets supplemented with different levels of VK3 (2.5, 5.0, 7.5, 10.0, and 12.5 mg/kg) for 11 wk. Dietary VK3 supplementation linearly and quadratically increased feed intake, egg mass, egg weight, and egg production (P < 0.05). Increasing VK3 levels linearly and quadratically increased albumen height, shell thickness and Haugh unit of eggs (P < 0.05). VK3 reduced osteocalcin (OC) and uncarboxylated osteocalcin (ucOC) levels in the serum. Dietary VK3 addition linearly decreased serum malondialdehyde (MDA) levels (P < 0.01). There was linear and quadratic effect in the activity of serum total superoxide dismutase (T-SOD) (P < 0.01), and linear effect in serum total antioxidant capacity (T-AOC) (P < 0.01). In conclusion, dietary VK3 supplementation enhanced the production performance, egg quality, vitamin K-dependent proteins, and antioxidant properties in breeding geese during the laying period. The optimal dose of dietary VK3 supplementation was 10.0 mg/kg.

Formulation improvement of a concentrated enzyme detergent for high-speed rail trains through life cycle assessment methodology.

High-speed rail has been operating in more than 25 countries (mainly in Asia, Europe and North America), and has become an important part of global economic development. However, the cleaning and maintenance of high-speed rail is a comprehensive task, which may easily cause environmental pollution. This study aims to analyze and improve the sustainability of the formulation and production process of a concentrated complex enzyme detergent used as the maintenance agent for high-speed trains via the life cycle assessment (LCA) method. The eFootprint software system with built-in China, European and Swiss Ecoinvent databases was used to establish the LCA model with the system boundary being from cradle to gate. The LCA model showed that the production of 1 kg of concentrated detergent generates the global warming potential of 2.53 kg CO2 eq, and other environmental emissions including acidification potential of 0.01 kg SO2 eq, eutrophication potential of 3.76E-03 kg PO43-eq, inhalable inorganic matter of 3.17E-03 kg PM2.5 eq, ozone depletion potential of 5.3E-06 kg CFC-11 eq, photochemical ozone formation potential of 3.44E-03 kg NMVOC eq, primary energy demand of 3.17 MJ, abiotic depletion potential of 4.97E-6 kg antimony eq, and water use of 0.84 kg. LCA results are not strongly dependent to the assumptions of the research, and the uncertainties of LCA results are between 8 and 16%, which is mainly due to the regional differences in technology sources, the year of technical data collection, and the representativeness of technology collection companies. Carbon footprint analysis showed that the production processes of enzyme stabilizer (glycerol) and surfactants contributed the most, while changes in power consumption during production and transportation distance of raw materials had limited effect on total carbon emissions. Therefore, the formulation of the concentrated complex enzymatic detergent was improved based on the LCA results. The new formulations with less enzyme stabilizer showed similar detergency to the original formulation. The new formulations could reduce carbon emissions by 5,500-9,200 tons per year and save between $4.4 and $7.4 million in annual production of 10,000 tons. Supplementary Information: The online version contains supplementary material available at 10.1007/s10668-023-03122-2.

Effects of dietary arginine supplementation on production performance, serum biochemicals, antioxidant capacity, and immunity of laying Wulong geese.

This study investigated the effects of dietary arginine supplementation on the production performance, serum biochemicals, antioxidant capacity, and immunity of laying Wulong geese. A total of 150 Wulong geese (34-wk old) with similar body weights were randomly divided into 6 groups with 5 replicates and 5 geese each (1 male and 4 female). The geese in the control group were fed a corn-rapeseed meal basal diet, and the geese in the treatment groups were fed the basal diet supplemented with 0.1, 0.2, 0.3, 0.4, and 0.5% arginine. The experiment lasted for 17 wk. Our results showed that dietary arginine increased the egg production rate (LR) and average egg weight (AEW) of geese in a quadratic manner (P < 0.05). Dietary arginine had a quadratic effect on the contents of total protein (TP) and triglyceride (TG) (P < 0.05) in the serum. Dietary arginine quadratically decreased the content of malondialdehyde (MDA) and increased the activity of total superoxide dismutase (T-SOD) (P < 0.05). Dietary arginine supplementation linearly and quadratically increased the contents of immunoglobulin A (IgA) and immunoglobulin G (IgG), and linearly increased the content of nitric oxide (NO) (P < 0.05). In conclusion, dietary arginine supplementation can significantly improve the production performance, serum biochemicals, antioxidant capacity, and immunity of laying Wulong geese. Therefore, 0.3% arginine (actual content: 1.02%) is recommended in the diet.

Novel Nonstoichiometric Niobium Oxide Anode Material with Rich Oxygen Vacancies for Advanced Lithium-Ion Capacitors.

Given the inherent features of open tunnel-like structures, moderate lithiation potential (1.0-3.0 V vs Li/Li+), and reversible redox couples (Nb5+/Nb4+ and Nb4+/Nb3+ redox couples), niobium-based oxides with Wadsley-Roth crystallographic shear structure are promising anode materials. However, their practical rate capability and cycling stability are still hindered by low intrinsic electronic conductivity and structural stability. Herein, ultrathin carbon-confined Nb12O29 materials with rich oxygen vacancies (Nb12O29-x@C) were designed and synthesized to address above-mentioned challenges. Computational simulations combined with experiments reveal that the oxygen vacancies can regulate the electronic structure to increase intrinsic electronic conductivity and reduce the Li+ diffusion barrier. Meanwhile, the carbon coating can enhance structural stability and further improve the electronic conductivity of the Nb12O29 material. As a result, the as-prepared Nb12O29-x@C exhibits high reversible capacity (226 mAh g-1 at 0.1 A g-1), excellent high-rate performance (83 mAh g-1 at 5.0 A g-1), and durable cycling life (98.1% capacity retention at 1.0 A g-1 after 3000 cycles). The lithium storage mechanism and structural stability of Nb12O29-x@C were also revealed by in situ X-ray diffraction (XRD), ex situ X-ray photoelectron spectroscopy (XPS), and ex situ Raman spectroscopy. When applied as the anode of lithium-ion capacitors (LICs), the as-built LIC achieves high energy density (72.4 Wh kg-1) within the voltage window of 0.01-3.5 V, demonstrating the practical application potential of the Nb12O29-x@C materials.

Cationic-potential tuned biphasic layered cathodes for stable desodiation/sodiation.

Sodium layered oxides generally suffer from deep-desodiation instability in P2 structure and sluggish kinetics in O3 structure. It will be great to design P2/O3 biphasic materials that bring the complementary merits of both structures. However, such exploration is hindered by the ambiguous mechanism of material formation. Herein, supported by theoretical simulations and various spectroscopies, we prove that P2/O3 biphasic structures essentially originate from the internal heterogeneity of cationic potential, which can be realized by constraining the temperature-driven ion diffusion during solid-state reactions. Consequently, P2/O3 biphasic Na0.7Ni0.2Cu0.1Fe0.2Mn0.5O2-δ with well-designed quaternary composition is successfully obtained, exhibiting much-improved rate capabilities (62 mAh g-1 at 2.4 A g-1) and cycling stabilities (84% capacity retention after 500 cycles) than its single-phase analogues. Furthermore, synchrotron-based diffraction and X-ray absorption spectroscopy are employed to unravel the underlying sodium-storage mechanism of the P2/O3 biphasic structure. This work presents new insights toward the rational design of advanced layered cathodes for sodium-ion batteries.

Quality changes of duck meat during thermal sterilization processing caused by microwave, stepwise retort, and general retort heating.

The quality changes of duck meat during thermal sterilization using microwave, stepwise retort and general retort heating were evaluated. Results showed that compared with stepwise retort and general retort, duck meat subjected to microwave showed significantly higher gumminess, chewiness, cohesiveness and resilience as well as glutamic acid, lysine and total amino acids. Low-field NMR revealed that the relative content of immobilized water after microwave and stepwise retort treatment was significantly higher than that after general retort treatment. The relative content of 1-octen-3-ol with characteristic mushroom aroma was significantly higher with microwave and stepwise retort heating than with general retort heating, while 2-pentyl-furan with poor taste was only detected with general retort heating. The muscle bundles subjected to microwave were neatly arranged, similar to those with no thermal sterilization. Overall, the meat quality after three thermal sterilization treatment was microwave > stepwise retort > general retort.

Fabrication and Characterization of Chicken- and Bovine-Derived Chondroitin Sulfate/Sodium Alginate Hybrid Hydrogels.

The physicochemical properties and microstructure of hybrid hydrogels prepared using sodium alginate (SA) and chondroitin sulfate (CS) extracted from two animal sources were investigated. SA-based hybrid hydrogels were prepared by mixing chicken- and bovine-derived CS (CCS and BCS, respectively) with SA at 1/3 and 2/3 (w/w) ratios. The results indicated that the evaporation water loss rate of the hybrid hydrogels increased significantly upon the addition of CS, whereas CCS/SA (2/3) easily absorbed moisture from the environment. The thermal stability of the BCS/SA (1/3) hybrid hydrogel was higher than that of CCS/SA (1/3) hybrid hydrogel, whereas the hardness and adhesiveness of the CCS/SA (1/3) hybrid hydrogel were lower and higher, respectively, than those of the BCS/SA (1/3) hybrid hydrogel. Low-field nuclear magnetic resonance experiments demonstrated that the immobilized water content of the CCS/SA (1/3) hybrid hydrogel was higher than that of the BCS/SA (1/3) hybrid hydrogel. FTIR showed that S=O characteristic absorption peak intensity of BCS/SA (2/3) was obviously higher, suggesting that BCS possessed more sulfuric acid groups than CCS. SEM showed that the hybrid hydrogels containing CCS have more compact porous microstructure and better interfacial compatibility compared to BCS.

Circadian clock disruptions link oxidative stress and systemic inflammation to metabolic syndrome in obstructive sleep apnea patients.

Objectives: Obstructive sleep apnea (OSA) is an independent risk factor for metabolic syndrome (MetS). Recent studies have indicated that circadian clock genes were dysregulated in OSA. In addition, it is clear that the impairment of circadian clocks drives the progression of MetS. Therefore, we hypothesized that circadian rhythm disruption links OSA with MetS. Methods: A total of 118 participants, who underwent polysomnography (PSG) and were diagnosed as healthy snorers (control, n = 29) or OSA (n = 89) patients based on the apnea-hypopnea index (AHI), were enrolled in the present study. General information, anthropometric data, blood biochemical indicators, clock gene expressions, and levels of oxidative and inflammatory indicators were collected, determined, and compared in all the participants. Results: We found that Brain and muscle aryl hydrocarbon receptor nuclear translocator-like protein 1 (Bmal1) and Differentiated embryo chondrocyte 1 (Dec1) were upregulated, while Period 1 (Per1) was reduced in OSA patients. In addition, these changing trends were closely associated with the hypoxia indicator of AHI and have a significant impact on the presence of MetS components, such as hyperglycemia (Dec1 and Per1, p < 0.05 and 0.001, respectively), hypertension (Bmal1 and Dec1, p < 0.001 and 0.01, respectively), hyperlipidemia (Dec1, p < 0.01), and obesity (Dec1, p < 0.05). Notably, expressions of Dec1 correlated with IR and predicted the presence of MetS in OSA patients. Finally, we also observed that Dec1 expression was interrelated with levels of both oxidative indicators and inflammatory biomarkers (IL-6) in OSA. Conclusion: This study concluded that circadian clock disruptions, especially Dec1, link OSA with MetS in an oxidative and inflammatory-related manner. Circadian clock Dec1 can be used as a specific biomarker (p < 0.001) and therapeutic target in OSA combined with Mets patients.

Degradation of pefloxacin by hybrid hydrodynamic cavitation with H2O2 and O3.

The degradation of toxic chemicals, antibiotics and other residues in organic wastewater has attracted much attention. Among various degradation technologies, hydrodynamic cavitation (HC) reactors have the advantage of being simple to operate. Through the combination of HC and other oxidants, the removal efficiency and energy efficiency of organic matter can be greatly improved, and the consumption of chemicals and the processing costs can be reduced. In this work, HC technology combined with oxidants was used to degrade pefloxacin (PEF), and the effect of different operating conditions on PEF degradation was investigated. The results indicated that the removal efficiency of PEF treated with HC alone was 84.9% under the optimal HC conditions of pH 3.3 and 120 min, which is much higher than that (35.5%) of pH 5.3. When co-treating the PEF solution with HC and H2O2 at 0.3 MPa and pH 5.3, the optimal molar ratio of PEF to H2O2 was 1:5, the highest PEF removal efficiency was 69.7%, and the synergy index (SI) was 4.4. When combining HC with O3, the PEF removal efficiency gradually elevated with increasing ozone addition. When the addition amount of ozone was 0.675 g/h, the removal efficiency of PEF was the highest, which was 91.5% after treatment of 20 min. The intermediate products in the reaction process were analyzed based on UV-Vis spectroscopy and LC-MS, and the mechanism and reaction pathways of PEF were proposed.

Effect of High-Voltage Electrostatic Field Heating on the Oxidative Stability of Duck Oils Containing Diacylglycerol.

High-voltage electrostatic field (HVEF) as an emerging green technology is just at the beginning of its use in meat products and by-products processing. In this study, we employed duck oil to produce duck-oil-based diacylglycerol (DAG), termed DDAG. Three different DDAG volume concentrations (0, 20%, and 100%) of hybrid duck oils, named 0%DDAG, 20%DDAG, and 100%DDAG, respectively, were used to investigate their thermal oxidation stability in high-voltage electrostatic field heating and ordinary heating at 180 ± 1 ℃. The results show that the content of saturated fatty acids and trans fatty acids of the three kinds of duck oils increased (p < 0.05), while that of polyunsaturated fatty acids decreased (p < 0.05) from 0 h to 8 h. After heating for 8 h, the low-field nuclear magnetic resonance showed that the transverse relaxation time (T21) of the three oils decreased (p < 0.05), while the peak area ratio (S21) was increased significantly (p < 0.05). The above results indicate that more oxidation products were generated with heating time. The peroxide value, the content of saturated fatty acids, and the S21 increased with more DAG in the duck oil, which suggested that the oxidation stability was likely negatively correlated with the DAG content. Moreover, the peroxide value, the content of saturated fatty acids and trans fatty acids, and the S21 of the three concentrations of duck oils were higher (p < 0.05) under ordinary heating than HVEF heating. It was concluded that HVEF could restrain the speed of the thermal oxidation reaction occurring in the duck oil heating and be applied in heating conditions.

Effects of heat treatment on physicochemical and microstructure properties of myofibrillar proteins combined with glucose and cellulose nanofibers.

The effects of unheated, 80 °C, 100 °C, and 121 °C on the physicochemical and microstructural properties of myofibrillary protein (MP) supplemented with glucose (Glc) and cellulose nanofibers (CNFs) were studied. The results showed that Glc and CNFs cross-linked with MP through non-covalent bonds when unheated compared with the unheated MP group, which increased the particle size and surface hydrophobicity of MP, and changed the secondary structure. At 80 °C and 100 °C, the particle size and hydrophobicity of MP significantly increased. At 121 °C, the contents of leucine and lysine in MP decreased by 16.2% and 19.9%, respectively, however, both Glc and CNFs could prevent this decrease. Meanwhile, microscopic images showed that the dispersion of Glc-MP and CNFs-MP were more uniform than the MP groups. Compared with Glc-MP, CNFs-MP had more uniform particle size distribution and better thermal stability at high temperatures (121 °C).

A comprehensive review on persulfate activation treatment of wastewater.

With increasingly serious environmental pollution and the production of various wastewater, water pollutants have posed a serious threat to human health and the ecological environment. The advanced oxidation process (AOP), represented by the persulfate (PS) oxidation process, has attracted increasing attention because of its economic, practical, safety and stability characteristics, opening up new ideas in the fields of wastewater treatment and environmental protection. However, PS does not easily react with organic pollutants and usually needs to be activated to produce oxidizing active substances such as sulfate radicals (SO4-) and hydroxyl radicals (OH) to degrade them. This paper summarizes the research progress of PS activation methods in the field of wastewater treatment, such as physical activation (e.g., thermal, ultrasonic, hydrodynamic cavitation, electromagnetic radiation activation and discharge plasma), chemical activation (e.g., alkaline, electrochemistry and catalyst) and the combination of the different methods, putting forward the advantages, disadvantages and influencing factors of various activation methods, discussing the possible activation mechanisms, and pointing out future development directions.

Exosomal MALAT1 Derived from High Glucose-Treated Macrophages Up-Regulates Resistin Expression via miR-150-5p Downregulation.

Metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) plays a crucial role in the pathophysiological process associated with diabetes-related complications. The effect of high glucose levels on macrophage-derived exosomal MALAT1 is unknown. Therefore, we investigated the molecular regulatory mechanisms controlling exosomal MALAT1 in macrophages under high glucose treatment and the therapeutic target of macrophage-derived exosomal MALAT1 using a balloon injury model of vascular disease in diabetic rats. High glucose (25 mM) significantly increased MALAT1 expression in macrophage-derived exosomes. MALAT1 suppressed miR-150-5p expression in macrophage-derived exosomes under high-glucose conditions. Silencing MALAT1 using MALAT1 siRNA significantly reversed miR-150-5p expression induced by macrophage-derived exosomes. Macrophage-derived exosomes under high-glucose treatment significantly increased resistin expression in macrophages. Silencing MALAT1 and overexpression of miR-150-5p significantly decreased resistin expression induced by macrophage-derived exosomes. Overexpression of miR-150-5p significantly decreased resistin luciferase activity induced by macrophage-derived exosomes. Macrophage-derived exosome significantly decreased glucose uptake in macrophages and silencing MALAT1, resistin or overexpression of miR-150-5p significantly reversed glucose uptake. Balloon injury to the carotid artery significantly increased MALAT1 and resistin expression and significantly decreased miR-150-5p expression in arterial tissue. Silencing MALAT1 significantly reversed miR-150-5p expression in arterial tissue after balloon injury. Silencing MALAT1 or overexpression of miR-150-5p significantly reduced resistin expression after balloon injury. In conclusion, high glucose up-regulates MALAT1 to suppress miR-150-5p expression and counteracts the inhibitory effect of miR-150-5p on resistin expression in macrophages to promote vascular disease. Macrophage-derived exosomes containing MALAT1 may serve as a novel cell-free approach for the treatment of vascular disease in diabetes mellitus.

Effects of zinc and Bacillus subtilis on the reproductive performance, egg quality, nutrient digestion, intestinal morphology, and serum antioxidant capacity of geese breeders.

The effects of zinc (Zn) and Bacillus subtilis (B. subtilis) on reproductive performance, egg quality, nutrient digestion, intestine morphology, and antioxidant capacity were explored in geese breeders. Geese breeders (n = 120, 46-wk of age) were randomly assigned into 6 groups with 4 replicates of 5 birds each (1 male and 4 female). Breeders were fed diets with 2 levels of B. subtilis (2.5 × 109 and 5 × 109 CFU/kg) crossed with three levels of Zn (25, 45, and 65 mg/kg) for duration of 10-wk. The results showed that the egg laying rate (P < 0.05), fertility rate (P < 0.01), hatchability rate (P < 0.05), yolk color (P < 0.05), and the retentions of crude protein (P < 0.05), ether extract (P < 0.05) and phosphorus of geese breeders were improved by dietary supplementation of 5 × 109 CFU/kg B. subtilis and 25 mg or 45 mg/kg Zn. The serum T-SOD (P < 0.05) was increased by 45 mg/kg Zn supplementation. The serum T-AOC (P < 0.05) and retention of Zn (P < 0.05) were increased by 5 × 109 CFU/kg B. subtilis supplementation. The birds fed with 5 × 109 CFU/kg B. subtilis and 25 mg or 45 mg/kg Zn showed improved villus length (P < 0.01) and villus length/ crypt depth (P < 0.01) in both the jejunum and ileum. In conclusion, the combination of B. subtilis and Zn may have synergistic effects on these parameters, and dietary inclusion of 5 × 109 CFU/kg B. subtilis and 45 mg/kg Zn is recommended for improving the reproductive performance of geese breeders.

[Current status and prospect of DNA synthesis in industrial biotechnology].

DNA synthesis is one of the most basic, widely-used tools in life science as well as a key enabling technology in synthetic biology. The rapid development of industrial biotechnology promoted by synthetic biology is creating an insatiable demand for large-scale DNA synthesis from more convenient, economical and safe sources. Industrial DNA synthesis platforms have remarkable advantages in terms of throughput, cost and speed. The research and development processes of industrial biotechnology benefit from these advantages, achieving a higher efficiency and lower cost. However, challenges in DNA manufacturing process remain, such as the use of large amounts of organic reagents, waste of resources and so on. With the continuous and rapid increase of DNA synthesis scale, the hazard of toxic chemicals, cost burden and environmental burden are becoming prominent. Based on our practical work on DNA synthesis, we discuss the demand and strategies for large-scale DNA synthesis in industrial biotechnology as well as the issues and potential solutions for sustainable development.