Calcium-peroxide-mediated cascades of oxygen production and glutathione consumption induced efficient photodynamic and photothermal synergistic therapy.

Photodynamic therapy (PDT) and photothermal therapy (PTT) are potent approaches to cancer treatment. However, the tumor microenvironment (TME) characterized by severe hypoxia and abundant glutathione (GSH) significantly reduces the effectiveness of PDT. In this study, we developed an oxidative stress amplifier CaO2/ICG@ZIF-8, which was capable of self-sufficient O2 delivery and GSH depletion to enhance PDT and PTT synergistic therapy. We utilized ZIF-8 as nanocarriers that when loaded with CaO2 and indocyanine green (ICG) form CaO2/ICG@ZIF-8 nanoparticles, which exhibit a uniform particle size distribution and a hydrated particle size of about 215 nm. CaO2 reacts with water under acidic conditions to produce O2 so CaO2/ICG@ZIF-8 has an excellent O2 supply capacity, which is essential for PDT. Moreover, CaO2/ICG@ZIF-8 also reacts with GSH to form glutathione disulfides (GSSH), enhancing the therapeutic outcome of PDT by preventing the consumption of local ractive oxygen species. Beyond that, CaO2/ICG@ZIF-8 can produce strong hyperthermia with a photothermal conversion efficiency of about 44%, which is exceedingly appropriate for PTT. Owing to its augmentation, PTT/PDT mediated by CaO2/ICG@ZIF-8 demonstrates intense tumor inhibitory effects in both in vitro and in vivo studies. Notably, the Zn and Ca generated by CaO2/ICG@ZIF-8 degradation are essential elements for the body, so CaO2/ICG@ZIF-8 shows favorable safety. Altogether, the research provides a promising PDT/PTT synergistic therapeutic strategy for cancer and may show more medical applications in the future.

Sulfotransferase 1C2 promotes hepatocellular carcinoma progression by enhancing glycolysis and fatty acid metabolism.

BACKGROUND: Hepatocellular carcinoma (HCC) is aggressive liver cancer. Despite advanced imaging and other diagnostic measures, HCC in a significant portion of patients had reached the advanced stage at the first diagnosis. Unfortunately, there is no cure for advanced HCC. As a result, HCC is still a leading cause of cancer death, and there is a pressing need for new diagnostic markers and therapeutic targets. METHODS: We investigated sulfotransferase 1C2 (SUTL1C2), which we recently showed was overexpressed in human HCC cancerous tissues. Specifically, we analyzed the effects of SULT1C2 knockdown on the growth, survival, migration, and invasiveness of two HCC cell lines, i.e., HepG2 and Huh7 cells. We also studied the transcriptomes and metabolomes in the two HCC cell lines before and after SULT1C2 knockdown. Based on the transcriptome and metabolome data, we further investigated the SULT1C2 knockdown-mediated shared changes, i.e., glycolysis and fatty acid metabolism, in the two HCC cell lines. Finally, we performed rescue experiments to determine whether the inhibitory effects of SULT1C2 knockdown could be rescued via overexpression. RESULTS: We showed that SULT1C2 overexpression promoted the growth, survival, migration, and invasiveness of HCC cells. In addition, SULT1C2 knockdown resulted in a wide range of gene expression and metabolome changes in HCC cells. Moreover, analysis of shared alterations showed that SULT1C2 knockdown significantly suppressed glycolysis and fatty acid metabolism, which could be rescued via SULT1C2 overexpression. CONCLUSIONS: Our data suggest that SULT1C2 is a potential diagnostic marker and therapeutic target for human HCC.

A novel autophagy-related long non-coding RNAs signature predicting progression-free interval and I-131 therapy benefits in papillary thyroid carcinoma.

This study aimed to explore the prognostic and predictive value of autophagy-related lncRNAs in papillary thyroid carcinoma (PTC). The expression data of autophagy-related genes and lncRNAs of the PTC patients were obtained from TCGA database. Autophagy-related-differentially expressed lncRNAs (DElncs) were identified and used to establish the lncRNAs signature predicting patients' progression-free interval (PFI) in the training cohort. Its performance was assessed in the training cohort, validation cohort, and entire cohort. Effects of the signature on I-131 therapy were also explored. We identified 199 autophagy-related-DElncs and constructed a novel six-lncRNAs signature was constructed based on these lncRNAs. This signature had a good predictive performance and was superior to TNM stages and previous clinical risk scores. I-131 therapy was found to be associated with favorable prognosis in patients with high-risk scores but not those with low-risk scores. Gene set enrichment analysis suggested that a series of hallmark gene sets were enriched in the high-risk subgroup. Single-cell RNA sequencing analysis suggested that the lncRNAs were mainly expressed in thyroid cells but not stromal cells. In conclusion, our study constructed a well-performed six-lncRNAs signature to predict PFI and I-131 therapy benefits in PTC.

Early Supplementation with Starter Can Improve Production Performance of Lambs but this Growth Advantage Disappears after 154 Days of Age.

The aim of this experiment was to study the sustained effects of early supplementation with a starter on the performance of lambs during the pre-weaning, post-weaning, and fattening periods. Sixty male Hu lambs (3.59 ± 0.05 kg) were randomly assigned to two (30 lambs per group) treatments. The lambs were fed milk replacer from three days of age. The early supplementation (ES) group was supplemented with a starter ration at seven days of age, the control (CON) group was supplemented at 21 days of age, and lambs in both groups were weaned from milk replacer at 28 days of age. Eight lambs from each group were randomly slaughtered at 98 days of age, and the remaining lambs were fed the same nutrient level of a fattening ration until slaughter at 200 days of age. The results showed that early supplementation with starter significantly improved average daily feed intake (ADFI) and average daily weight gain (ADG) in the pre-weaning period (7-28 days of age), and ADFI and slaughter performance (live weight before slaughter, carcass weight and dressing percentage) in the post-weaning period (29-98 days of age, p < 0.05). In addition, early supplementation with the starter had no significant effect (p > 0.05) on ADFI, ADG, and slaughter performance, but significantly decreased (p < 0.05) the feed-to-gain ratio (F/G) of lambs during the fattening period (99-200 days of age). In addition, early supplementation with starter increased the ratio of rumen and reticulum weight to total stomach weight in lambs (p < 0.05). In conclusion, early supplementation with starter can reduce feed costs and improve the performance of lambs, while the growth advantage produced by early supplementation had initially disappeared by 154 days of age.

Multi-Disaster Integrated Risk Assessment in City Range-A Case Study of Jinan, China.

Urban multi-disaster integrated risk assessment is an important part of urban sustainable development and territorial spatial planning. Based on the results of integrated risk assessment, the scientific and effective performance of disaster prevention and reduction can be effectively improved. This study determines a multi-disaster integrated risk assessment system. The system evaluates the hazard level of disasters, the exposure level of disaster bearing bodies, the vulnerability level of disaster bearing bodies, and the urban resilience level, and determines the city's integrated risk level on this basis. Taking Jinan as an example, the risk, exposure, vulnerability, resilience, and integrated risk level of Jinan City were analyzed. The results show that the system reasonably analyzes the multi-disaster integrated risk level, and according to the assessment results, countermeasures for disaster prevention and suggestions for territorial spatial planning were put forward.

miR-21 Negatively Regulates the PTEN-PI3K-Akt-mTOR Signaling Pathway in Crohn's Disease by Altering Immune Tolerance and Epithelial-Mesenchymal Transition.

miR-21 is involved in the mechanisms of inflammatory bowel disease (IBD). It negatively regulates PTEN, which is an upstream regulatory gene of the PI3K/Akt/mTOR signaling pathway. But the relationship between miR-21 and immune tolerance and intestinal epithelial injury, and the mechanism by which miR-21 participates in Crohn's disease (CD) have not been studied. We aimed to address these two questions. The results of the present study showed that the levels of miR-21 and PTEN respectively decreased and increased significantly in the intestinal mucosa from active CD compared with control ones. Transfection with miR-21-5p mimics significantly downregulated the expression of PTEN and upregulated PI3K-Akt-mTOR signaling and the downstream pathway in PBMCs, while transfection with a miR-21-5p inhibitor antagomiR-21had the opposite effect. Moreover, the ratio of Treg/Th1 cells differentiated from peripheral blood mononuclear cells (PBMCs) decreased after being transfected with mimics, and increased with the inhibitor. AntagomiR-21 significantly relieved the lesions in colons of mice with TNBS-induced colitis, accompanied by the upregulation of PTEN and downregulation of mTOR. Inhibition of miR-21 also effectively suppressed the process of epithelial-mesenchymal transition (EMT) in vivo. In conclusion, the level of miR-21 decreased in CD, resulting in an upregulated PI3K-Akt-mTOR signaling pathway, compromised immune tolerance, and elevated inflammation.

Study on the Solidification Effect of Dredger Fill by Microbial-Induced Calcium Precipitation (MICP).

This paper puts forward a new soft soil reinforcement technology-microbial-induced calcite precipitation (MICP) technology-which considers the problem of dredger fill soft-soil reinforcement in Dalian Taiping Bay. In this paper, the calcium carbonate content (CCC) and unconfined compressive strength (UCS) of microbial solidified dredger fill (MSDF) samples were determined using laboratory experiments. The microstructure and chemical composition of MSDF samples were studied by SEM-EDS and XRD. The failure and reinforcement mechanism of MSDF under different experimental conditions (ambient temperature, cementation solution concentration, and clay content) were investigated. The results showed that there was a certain residual strength after the peak strength of MSDF. With the increase of ambient temperature, the number of microorganisms increased, but the activities of urease, CCC, and UCS decreased. The UCS and CCC increased with the increase of cementation solution concentration, while they first increased and then decreased with the increase of clay content. The clay content enhanced the compactness of MSDF samples but reduced the soil permeability and weakened the mineralization. There were significant differences in the morphology of microbial-induced precipitation caused by different concentrations of cementation solution.

Numerical simulation of closed plastic impeller molding process and its parameter optimization.

Aiming at the warping deformation and volume shrinkage of the closed plastic impeller, the relevant compression molding process parameters were optimized.Based on the theoretical equation of the pressing pressure for general sheet plastics, the theoretical derivation of the pressing pressure suitable for cylindrical cavities was carried out, and the theoretical derivation of the pressing pressure suitable for closed plastic impeller molding was derived, and the pressing pressure was calculated to be 15 MPa. The warpage deformation and shrinkage rates under different combinations of process parameters were obtained, and the influence of process parameters on warpage deformation and shrinkage rates, as well as the two combinations of process parameters that satisfy the best warpage deformation and shrinkage rates, respectively, were obtained by extreme difference analysis. A GA-BP neural network model was established for the prediction of the process parameters of closed plastic impeller compression molding, and the prediction curves were fitted into a function to obtain a set of process parameter combinations with optimal warpage and shrinkage at the same time by using the multi-objective optimization function of NSGA-II algorithm.

The economic impact of porcine reproductive and respiratory syndrome outbreak in four Chinese farms: Based on cost and revenue analysis.

The economic impact after the outbreak of porcine reproductive and respiratory syndrome (PRRS) has been proven to be tremendous for pig production worldwide. However, the economic impact of the disease is not well understood in China. In our previous study, we acquired and analyzed the main production data (the number of weaned piglets, health costs, delayed marketing age, etc.) from the management system before and after the PRRS outbreaks occurring in November 2014, March 2015, December 2016, and February 2017. This study aimed to analyze and quantify the economic losses of the four PRRS outbreaks in Chinese herds. A straightforward approach was used to calculate additional costs and decreased revenues based on the PRRS-induced production deficiencies by average cost-of-production indices calculated from annual estimates of costs between 2014 and 2017. The results showed that economic losses varied between ¥668.14 and ¥1004.43 per sow in breeding herds from the outbreaks to regain the basic performance, with an average of ¥822.75 per sow, and the mean costs in the fattening herds (including nursery pigs) were ¥601.62 per sow, ranging from ¥318.64 to ¥937.14. Overall, the economic impact of PRRS on the whole herd was ¥1424.37 per sow. The majority of the losses were due to the reduction in the number of weaned piglets for breeding herds, and the increased feed cost (occupying 44.88%) was the primary source of loss for fattening herds. Our study fills the gap in knowledge of PRRS economics in China, enriches the data for veterinary economics, and re-stresses the necessity for producers and veterinarians to control PRRS effectively.

Accelerated degradation of cellulose in silkworm excrement by the interaction of housefly larvae and cellulose-degrading bacteria.

The environmental pollution caused by silkworm (Bombyx mori) excrement is prominent, and rich in refractory cellulose is the bottleneck restricting the efficient recycling of silkworm excrement. This study was performed to investigate the effects of housefly larvae vermicomposting on the biodegradation of cellulose in silkworm excrement. After six days, a 58.90% reduction of cellulose content in treatment groups was observed, which was significantly higher than 11.5% of the control groups without housefly larvae. Three cellulose-degrading bacterial strains were isolated from silkworm excrement, which were identified as Bacillus licheniformis, Bacillus amyloliquefaciens, and Bacillus subtilis based on 16S rRNA gene sequence analysis. These three bacterial stains had a high cellulose degradation index (HC value ranged to between 1.86 and 5.97 and FPase ranged from 5.07 U/mL to 7.31 U/mL). It was found that housefly larvae increased the abundance of cellulose-degrading bacterial genus (Bacillus and Pseudomonas) by regulating the external environmental conditions (temperature and pH). Carbohydrate metabolism was the bacterial communities' primary function during vermicomposting based on the PICRUSt. The results of Tax4Fun indicated that the abundance of endo-ß-1,4-glucanase and exo-ß-1,4-glucanase increased rapidly and maintained at a higher level in silkworm excrement due to the addition of housefly larvae, which contributed to the accelerated degradation of cellulose in silkworm excrement. The finding of this investigation showed that housefly larvae can significantly accelerate the degradation of cellulose in silkworm excrement by increasing the abundance of cellulose-degrading bacterial genera and cellulase.

Impact of Coastal Urbanization on Marine Pollution: Evidence from China.

The increasing marine pollution in China's coastal areas has seriously affected the sustainable development of the economy and the living standards of residents. It is of great significance to explore the relationship between urbanization and marine pollution in coastal areas for the sustainable development of coastal cities. Based on the marine pollution data and nighttime light (NTL) data of 46 coastal cities from 2006 to 2015, the paper discusses the impact of urbanization on marine pollution by using the generalized spatial two-stage least square method (GS2SlS), and analyzes the role of technological innovation, financial development, and human capital in the impact of urbanization on marine pollution by using the three-stage least square method (3SLS). Results show that China's coastal marine pollution has a strong spatial spillover effect, and a U-shaped relationship exists between urbanization and marine pollution. Regional heterogeneity analysis shows that an inverted U-shaped relationship was found between coastal urbanization and marine pollution in the northern marine economic circle, while the eastern and southern marine economic circles have a U-shaped correlation. The heterogeneity of the urbanization pattern indicates that the relationship between different urbanization patterns and marine pollution in coastal areas is generally in a positive correlation stage, but the depth of urbanization occupies a dominant position. Further mechanism tests show that urbanization can effectively reduce coastal marine pollution and improve the marine environment through the technological innovation effect, financial development effect, and human capital effect.

A New Parameter of Hip Instability in Developmental Dysplasia of the Hip (DDH): Teardrop Distance.

Background: Hip instability is one of the etiologies of accelerated onset of osteoarthritis in developmental dysplasia of the hip (DDH). There are some radiological parameters for hip instability in hip dysplasia like broken shenton's line, elevated acetabular index, reduced lateral center edge angle (LCEA), upsloping lateral sourcil. We have discovered a new index of teardrop distance (TD) for assessing instability. Herein, we hypothesized that increased TD could be used as evidence of hip instability in DDH patients, which we verified using TD as an auxiliary diagnostic parameter for DDH, from supine to standing position. Methods: Female DDH patients undergoing Bernese periacetabular osteotomy (PAO) were enrolled in the DDH group, and normal female volunteers were in the control group. Anteroposterior radiographs of the pelvis in the supine and standing positions were taken, and LCEA, Tönnis angle (TA), sharp angle (SA), and TD were tested using Stata software to analyze the changes between supine and standing anteroposterior pelvic radiographs. Results: There were 26 female volunteers with 52 hips in the control group: supine TD 6.80 ± 0.98 mm, standing TD 6.65 ± 1.3 mm (P > 0.05). A total of 78 patients with 135 hips were included in the DDH group: supine TD 10.51 ± 3.50 mm, standing TD 10.93 ± 4.23 mm (P < 0.05). In either supine or standing position, TD in the DDH group was significantly wider than that in the control group (P < 0.05). In the DDH group, TD was correlated with TA and LCEA (rp 0.494-0.588, P < 0.05); TD was not correlated with SA, weight, or BMI (P > 0.05). There was a weak correlation between TD difference and standing LCEA (rp -0.276, P < 0.05). Conclusion: TD > 10 mm was a common imaging feature of DDH. It increased from supine to standing position, thus indicating hip instability in DDH patients. The hip parameters of both positions should be compared, fully considering the factors of hip stability.

Aspirin eugenol ester alleviates lipopolysaccharide-induced acute lung injury in rats while stabilizing serum metabolites levels.

Aspirin eugenol ester (AEE) was a novel drug compound with aspirin and eugenol esterified. AEE had various pharmacological activities, such as anti-inflammatory, antipyretic, analgesic, anti-oxidative stress and so on. In this study, it was aimed to investigate the effect of AEE on the acute lung injury (ALI) induced by lipopolysaccharide (LPS) in rats. In vitro experiments evaluated the protective effect of AEE on the LPS-induced A549 cells. The tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1ß (IL-1ß) were measured in the cell supernatant. The Wistar rats were randomly divided into five groups (n = 8): control group, model group (LPS group), LPS + AEE group (AEE, 54 mg·kg-1), LPS + AEE group (AEE, 108 mg·kg-1), LPS + AEE group (AEE, 216 mg·kg-1). The lung wet-to-dry weight (W/D) ratio and immune organ index were calculated. WBCs were counted in bronchoalveolar lavage fluid (BALF) and total protein concentration was measured. Hematoxylin-Eosin (HE) staining of lung tissue was performed. Glutathione (GSH), glutathione peroxidase (GPx), catalase (CAT), antioxidant superoxide dismutase (SOD), total antioxidant capacity (T-AOC), lactate dehydrogenase (LDH), C-reactive protein (CRP), myeloperoxidase (MPO), malondialdehyde (MDA), macrophage mobility inhibitory factor (MIF), TNF-α, IL-6, and IL-1ß activity were measured. The metabolomic analysis of rat serum was performed by UPLC-QTOF-MS/MS. From the results, compared with LPS group, AEE improved histopathological changes, reduced MDA, CRP, MPO, MDA, and MIF production, decreased WBC count and total protein content in BALF, pro-inflammatory cytokine levels, immune organ index and lung wet-dry weight (W/D), increased antioxidant enzyme activity, in a dose-dependent manner. The results of serum metabolomic analysis showed that the LPS-induced ALI caused metabolic disorders and oxidative stress in rats, while AEE could ameliorate it to some extent. Therefore, AEE could alleviate LPS-induced ALI in rats by regulating abnormal inflammatory responses, slowing down oxidative stress, and modulating energy metabolism.

Exogenous Melatonin Improves Seed Germination of Wheat (Triticum aestivum L.) under Salt Stress.

Melatonin (MT) can effectively reduce oxidative damage induced by abiotic stresses such as salt in plants. However, the effects of MT on physiological responses and molecular regulation during wheat germination remains largely elusive. In this study, the response of wheat seeds to MT under salt stress during germination was investigated at physiological and transcriptome levels. Our results revealed that application of MT significantly reduced the negative influence of salt stress on wheat seed germination. The oxidative load was reduced by inducing high activities of antioxidant enzymes. In parallel, the content of gibberellin A3 (GA3) and jasmonic acid (JA) increased in MT-treated seedling. RNA-seq analysis demonstrated that MT alters oxidoreductase activity and phytohormone-dependent signal transduction pathways under salt stress. Weighted correlation network analysis (WGCNA) revealed that MT participates in enhanced energy metabolism and protected seeds via maintained cell morphology under salt stress during wheat seed germination. Our findings provide a conceptual basis of the MT-mediated regulatory mechanism in plant adaptation to salt stress, and identify the potential candidate genes for salt-tolerant wheat molecular breeding.

Low-Velocity Impact Behavior of Sandwich Plates with FG-CNTRC Face Sheets and Negative Poisson's Ratio Auxetic Honeycombs Core.

The combination of auxetic honeycomb and CNT reinforcement composite is expected to further improve the impact protection performance of sandwich structures. This paper studies the low-velocity impact response of sandwich plates with functionally graded carbon nanotubes reinforced composite (FG-CNTRC) face sheets and negative Poisson's ratio (NPR) auxetic honeycomb core. The material properties of FG-CNTRC were obtained by the rule of mixture theory. The auxetic honeycomb core is made of Ti-6Al-4V. The governing equations are derived based on the first-order shear deformation theory and Hamilton's principle. The nonlinear Hertz contact law is used to calculate the impact parameters. The Ritz method with Newmark's time integration schemes is used to solve the response of the sandwich plates. The (20/-20/20)s, (45/-45/45)s and (70/-70/70)s stacking sequences of FG-CNTRC are considered. The effects of the gradient forms of FG-CNTRC surfaces, volume fractions of CNTs, impact velocities, temperatures, ratio of plate length, width and thickness of surface layers on the value of the plate center displacement, the recovery time of deformation, contact force and contact time of low-velocity impact were analyzed in detail.

[Isolation, identification and biodegradation characterization of a novel monensin-degrading bacterial strain].

Biodegradation of antibiotic pollutants by microorganisms has received widespread attention, to which the identification of microorganisms capable of efficiently degrading antibiotics is a key. In this study, a strain DM-1 with high degradation capability was successfully isolated from monensin-contaminated chicken manure by using monensin as the sole carbon source. The strain was further identified basing on morphological, physiological and biochemical characteristics and 16S rRNA gene sequence-based phylogenetic analysis. The degradation efficiency of DM-1 for monensin was determined by HPLC post-column derivatization, and then the degradation conditions of DM-1 were optimized. DM-1 was identified as a strain of Acinetobacter and named as Acinetobacter baumannii DM-1. The optimal conditions for monensin degradation by strain DM-1 were pH 7.0, 30 ℃, and initial monensin concentration of 50 mg/L. The strain DM-1 degraded more than 87.51% of monensin at an initial concentration of 10 mg/L in 28 days, while only a slight decrease in monensin concentration was observed in the control without monensin-degrading strain. This study indicates that the strain DM-1 has a promising application prospect in the bioremediation of monensin-contaminated environment.

Analysis of bacterial diversity and genetic evolution of Lacticaseibacillus paracasei isolates in fermentation pit mud.

AIMS: Since little is known about the genetic diversity of lactic acid bacteria (LAB) isolates from the fermentation pit mud (FPM), we sought to evaluate the bacterial structure, identify the LAB isolates and investigate the genotype and genetic diversity of the LAB isolates. METHODS AND RESULTS: Using high-throughput MiSeq sequencing, we identified seven dominant bacterial genera in FPM. Lactobacillus had the highest abundance. We isolated 55 LAB strains. These isolates were all identified as Lacticaseibacillus paracasei. Using an extant multilocus sequence typing (MLST) scheme, isolates were assigned to 18 sequence types (STs) and three clonal complexes. ST1, the largest group, mainly comprised FPM isolates. Niche-specific ST2 to ST18 only contained FPM isolates. Isolates could be divided into four lineages, with most assigned to Lineage 1. Only one FPM isolate was classified as L. paracasei subsp. paracasei. Other isolates could not be classified at the subspecies level using the seven MLST loci. CONCLUSIONS: Lactobacilli account for a high proportion of bacteria in pit mud. Based on the traditional culture method, L. paracasei was the dominant species, and these isolates exhibit a high ethanol tolerance, high intraspecific diversity and specific genetic profiles. SIGNIFICANCE AND IMPACT OF THE STUDY: The study described the characterization of FPM bacterial diversity, giving an insight into the genetic diversity of L. paracasei strains present in FPM.

Characterization of two novel pentose-fermenting and GABA-producing species: Levilactobacillus tujiorum sp. nov. and Secundilactobacillus angelensis sp. nov. Isolated from a solid-state fermented zha-chili.

Lactobacilli are dominant in zha-chili. This study provides a taxonomic characterization of five bacterial strains isolated from zha-chili in China. The cells were Gram-positive, facultative anaerobic, non-spore-forming, flagella-free, catalase-negative, heterofermentative, pentose-fermenting, and gamma-aminobutyric acid (GABA)-producing rods. For HBUAS51241T, HBUAS51329, and HBUAS51416, C16:0, C18:1ω9c and C19:0 iso were the predominant cellular fatty acids; diphosphatidylglycerol (DPG), phosphatidylglycerol (DP), glycolipids (GL), and glycolipids (AL) were the major phospholipids. While for HBUAS51383T and HBUAS58055, C16:0, C18:1ω9c, C19:0 cyclo ω8c were the predominant cellular fatty acids; DPG, DP, GL, and AL were the major phospholipids. Strains HBUAS51241T, HBUAS51329, and HBUAS51416 showed 98.1-99.1% 16S rRNA gene sequence similarity, 80.2-81.4% ANI, 87.7-90.0% AAI, and 23.8-32.8% digital DDH to their closest related type strains Levilactobacillus hammesii DSM 16381T, Levilactobacillus parabrevis ATCC 53295T, and Levilactobacillus fuyuanensis 244-4T. Strains HBUAS51383T and HBUAS58055 showed 98.7-99.5% 16S rRNA gene sequence similarity, 75.4-81.4% ANI, 75.5-89.1% AAI, and 19.7-24.0% digital DDH to their closest related type strains Secundilactobacillus silagincola IWT5T, Secundilactobacillus silagei JCM 19001T, Secundilactobacillus pentosiphilus IWT25T, Secundilactobacillus mixtipabuli IWT30T, Secundilactobacillus odoratitofui DSM 19909T, and Secundilactobacillus similis DSM 23365T. The central carbon metabolism pathways for the five strains were summarizeded. Based on the phenotypic, chemotaxonomic, and genomic data, we propose two novel species Levilactobacillus tujiorum sp. nov. whose type strain is HBUAS51241T (=GDMCC 1.3022T = JCM 35241T), and Secundilactobacillus angelensis sp. nov. whose type strain is HBUAS51383T (=GDMCC 1.3021T = JCM 35209T).

Metagenomic and physicochemical analyses reveal microbial community and functional differences between three types of low-temperature Daqu.

Complex microbes of different types of low-temperature Daqu (LTD) play an important role in the formation of flavors and qualities of light-flavor Baijiu during fermentation. However, characterizing the taxonomic and functional diversity of microbiota in three types of LTD (Houhuo, Hongxin, Qingcha) remains a major challenge. The present study combined metagenomic sequencing with culture-based methods and physicochemical analysis to compare the three LTD microbiota and elucidate their function in LFB brewing. The results revealed a high diversity of microbes in LTD, with 1286 genera and 4157 species detected across all studied samples. Bacteria and fungi were the main microbes in LTD, with a bacterial to fungal relative abundance ratio of above 4:1. Bacillus (21.18%) and Bacillus licheniformis (17.45%) were the most abundant microbes in the LTD microbiota at the genus and species levels, respectively. Culture-dependent analysis found the highest abundances of bacteria, fungi, and lactic acid bacteria in Houhuo, while the metagenomic-based microbiota found that the relative abundance of bacteria and fungi were highest in Houhuo and Hongxin among the three types of LTD, respectively. The different production temperatures of LTD had little effect on its microbial variety, but obviously impacted the microbiota structure and metagenomic function of LTD. Although the microbiota of the three types of LTD shared a high commonality, each had specific microbiota and functional metagenomic features, suggesting their different but complementary roles in the LFB fermentation process. The representative dominant microbes in Houhuo were mostly involved in metabolic pathways associated with the production of flavor substances in liquor. In contrast, the enriched microbes in Qingcha and Hongxin were not only capable of producing specific flavor substances but also had a strong ability to degrade macromolecular substances in raw materials, promoting microbial growth. This study has greatly enriched our knowledge of the effect of LTD fermentation temperature on its quality, providing practical and interesting information for future improvement of LTD and light-flavor Baijiu products.

Uptake and Transport of Naringenin and Its Antioxidant Effects in Human Intestinal Epithelial Caco-2 Cells.

Naringenin, a flavanone, has been reported for a wide range of pharmacological activities. However, there are few reports on the absorption, transport and antioxidant effects of naringenin. The study was to explore the uptake, transport and antioxidant effects of naringenin in vitro. Cell transmembrane resistance, lucifer yellow transmission rate, and alkaline phosphatase activity were used to evaluate the successful construction of cell model. The results showed that the absorption and transport of naringenin by Caco-2 cells were time- and concentration-dependent. Different temperatures (37 and 4°C) had a significant effect on the uptake and transport of naringenin. Verapamil, potent inhibitor of P-glycoprotein, significantly inhibit naringenin transport in Caco-2 cells. The results revealed that naringenin was a moderately absorbed biological macromolecule and can penetrate Caco-2 cells, mainly mediated by the active transport pathway involved in P-glycoprotein. At the same time, naringenin pretreatment could significantly increase the viability of H2O2-induced Caco-2 cells. Twenty four differential metabolites were identified based on cellular metabolite analysis, mainly including alanine, aspartate and glutamate metabolism, histidine metabolism, taurine and hypotaurine metabolism, pyruvate metabolism, purine metabolism, arginine biosynthesis, citrate cycle, riboflavin metabolism, and D-glutamine and D-glutamate metabolism. We concluded that the transport of naringenin by Caco-2 cells is mainly involved in active transport mediated by P-glycoprotein and naringenin may play an important role in oxidative stress-induced intestinal diseases.