Degradation and transformation mechanisms of pungent substances in huajiao (Zanthoxylum bungeanum) oil during storage: Induced by ultraviolet irradiation.

The pungency of huajiao (scientifically known as Zanthoxylum bungeanum) oil (ZBO), a crucial seasoning oil, is notably influenced by storage conditions, an aspect insufficiently explored in current research. Through the use of high-performance liquid chromatography and liquid chromatography-mass spectrometry, this study systematically investigated the stability of pungent compounds in ZBO under various storage conditions. It also elucidated the degradation and transformation mechanisms of these substances when exposed to ultraviolet (UV) irradiation. The results underscore elevated temperature, light exposure, oxygen, and storage duration as pivotal factors influencing compound degradation, with UV light emerging as the primary driving force. After 48 h of UV exposure, the primary pungent compound, hydroxy-α-sanshool, experienced a significant loss of 85.49%, indicating a pronounced inclination towards isomerization and oxidation. Notably, this study reveals, for the first time, the possible degradation-transformation pattern of hydroxy-γ-sanshool: a mutual conversion with hydroxy-γ-isosanshool and isomerization to (2E,4E,8Z,10E,12Z)-N-(2-hydroxy-2-methylpropyl) tetradeca-2,4,8,10,12-pentaenamide.

Postoperative rehabilitation and quality of life evaluation for transoral endoscopic thyroidectomy vestibular approach.

There are no targeted rehabilitation training modalities and assessment tools for patients after transoral endoscopic thyroidectomy vestibular approach (TOETVA). Herein, we develop a new assessment questionnaire and rehabilitation training modality and evaluate its safety and effectiveness. The THYCA-QoL-TOETVA questionnaire was compiled, and reliability and validity analyses were performed. Patients were divided into the new rehabilitation training group (N) or the conventional rehabilitation training group (C), and 1:1 propensity score matching (PSM) was performed after administering questionnaires to patients in both groups. Cervical range of motion (CROM) data were also measured and collected for statistical analysis. The questionnaire used in this study showed good expert authority, coordination, internal consistency, and questionnaire reliability. A total of 476 patients were included after PSM, and the questionnaire results showed that recovery and quality of life were better in the N group than in the C group (124.55 ± 8.171 vs. 122.94 ± 8.366, p = 0.026). Analysis of cervical spine mobility showed that rehabilitation was better in the N group compared to the C group at postoperative one month (flexion: 1.762°, extension: 4.720°, left lateral bending: 3.912°, right lateral bending: 4.061°, left axial rotation: 5.180°, right axial rotation: 5.199°, p value all of these < 0.001), and at postoperative three months (flexion: 2.866°, extension: 2.904°, left lateral bending: 3.927°, right lateral bending: 3.330°, left axial rotation: 4.395°, right axial rotation: 3.992°, p value all of these < 0.001). The THYCA-QoL-TOETVA provides an appropriate and effective tool for measuring the postoperative quality of life of TOETVA patients. This new rehabilitation training can effectively alleviate the problem of limited neck movement and improve the quality of life of patients after TOETVA surgery.Trial registration: ChiCTR2300069097.

MUC4 O-GlcNAcylation Regulates the Epithelial Phenotype in Conjunctival Epithelial Cells.

In the present study, our aim was to explore the role of MUC4 in IL-4-stimulated conjunctival epithelial cells and the underlying mechanisms. Human recombinant IL-4 was employed in human conjunctival epithelial cells (HConEpic) cells, and MUC4 shRNA (sh-MUC4) was constructed to explore the functional role of MUC4. The protein level of MUC4, O-GlcNAc transferase (OGT), O-GlcNAc hydrolase (OGA), zonula occludens 1 (ZO-1), gap junction protein beta 2 (GJB2), claudin-8 (CLDN8), and E-cadherin were detected by Western blot in HConEpic cells, the interaction between MUC4 and OGT/OGA was assessed by co-immunoprecipitation (IP) and Western blot in 293T cells. Our results showed that IL-4 significantly up-regulated MUC4 and OGT protein levels in HConEpic cells, while down-regulated OGA protein level. Also, IL-4 down-regulated ZO-1, GJB2, CLDN8, and E-cadherin protein levels in HConEpic cells, while which was markedly reversed by sh-MUC4. Additionally, OGT inhibitor significantly reduced MUC4 protein level, and elevated ZO-1, GJB2, CLDN8, and E-cadherin protein levels in HConEpic cells, while OGA inhibitor resulted in the opposite results. Furthermore, in addition to the interaction between OGT/OGA and MUC4, Co-IP and Western blot also revealed the alteration of MUC4 O-GlcNAcylation in 293T cells treated with OGT/OGA inhibitor. Above findings suggested that OGT/OGA inhibitor regulated MUC4 protein level by affecting MUC4 O-GlcNAcylation to regulate ZO-1, GJB2, CLDN8, and E-cadherin protein levels in HConEpic cells, which was achieved via inhibiting the interaction between OGT/OGA and MUC4. This study may provide a better understanding of the pathogenesis of allergic conjunctivitis (AC).

Impact of aroma-enhancing microorganisms on aroma attributes of industrial Douchi: An integrated analysis using E-nose, GC-IMS, GC-MS, and descriptive sensory evaluation.

In order to enhance the aromatic profile of industrial Douchi, a comprehensive investigation was undertaken to assess the impact of aroma-enhancing microorganisms on the sensory attributes of Douchi. This evaluation utilized a combination of analytical techniques, including electronic nose analysis, gas chromatography-ion mobility spectrometry (GC-IMS), gas chromatography-mass spectrometry (GC-MS), and descriptive sensory analysis (DA). Both GC-IMS and GC-MS revealed significant changes in the volatile composition of Douchi following the addition of aroma-enhancing microorganisms (p < 0.05). Partial least squares-discriminant analysis (PLS-DA) identified benzaldehyde, benzene acetaldehyde, 3-octanone, and ethyl 2-methylbutyrate as significant differentiating volatile compounds. Additionally, compared to the control group, the sensory attributes of sourness in Douchi were significantly reduced (p < 0.001), while the attributes of wine-like and sweetness were notably enhanced (p < 0.05) when the ratio of G. candidum to C. versatilis was 1:1 (GCC group). By calculating the odor-activity values (OAVs) of key volatiles, it can be hypothesized that this aroma improvement of Douchi may be attributed to an increase in the typical volatiles (3-methyl-1-butanol, 1-octen-3-ol, 3-octanol, and 3-octanone) and ethyl 2-methylbutanoate with high OAVs (2340849.64 âˆ¼ 16695327.86), as well as to decreases in the musty acetophenone. In conclusion, the aroma profile of Douchi was significantly enhanced when G. candidum and C. versatilis were added at a ratio of 1:1. This study provides valuable insights into the development of aroma enhancers for improving the sensory profile of Douchi.

Virus-host coevolutionary analyses of an Alphabaculovirus with a wide host range.

Baculoviruses are highly host specific, and their host range is usually restricted to a single or a few closely related insect species, except for few virus species, e.g. Alphabaculovirus aucalifonicae and Alphabaculovirus mabrassicae. In this study, two new alphabaculovirus isolates were isolated from the larvae of Mamestra brassicae and Mythimna separata, which were named as Mamestra brassicae multiple nucleopolyhedrovirus isolate QD (MbMNPV-QD) and Mythimna separata multiple nucleopolyhedrovirus isolate Hb (MyseMNPV-Hb), respectively. The Kimura two-parameter values based on the concatenated 38 core genes of baculovirus revealed that MbMNPV (isolates QD/CHb1/K1/CTa), MyseMNPV-Hb, Helicoverpa armigera multiple nucleopolyhedrovirus (HearMNPV) and Mamestra configurata nucleopolyhedrovirus B (MacoNPV-B) were different isolates of a same virus species. A phylogenetic tree of baculoviruses and nudiviruses constructed from their 20 homologous gene sequences, and that of their isolated hosts constructed from 13 protein-coding genes of the insect mitochondrial genomes, were used to analyse the coevolution of baculoviruses with their isolated hosts. The results showed that M. brassicae was the most likely ancestral host of these virus isolates, included MbMNPV isolates, MyseMNPV-Hb, HearMNPV, and MacoNPV-B. Therefore, we concluded that these virus isolates belong to the existing virus species - Alphabaculovirus mabrassicae with M. brassicae as their ancestral host.

Causal associations between gut microbiota, gut microbiota-derived metabolites, and cerebrovascular diseases: a multivariable Mendelian randomization study.

Background: Mounting evidence has demonstrated the associations between gut microbiota, gut microbiota-derived metabolites, and cerebrovascular diseases (CVDs). The major categories of CVD are ischemic stroke (IS), intracerebral hemorrhage (ICH), and subarachnoid hemorrhage (SAH). However, the causal relationship is still unclear. Methods: A two-sample Mendelian randomization (MR) study was conducted leveraging the summary data from genome-wide association studies. The inverse variance-weighted, maximum likelihood, weighted median, and MR.RAPS methods were performed to detect the causal relationship. Several sensitivity analyses were carried out to evaluate potential horizontal pleiotropy and heterogeneity. Finally, reverse MR analysis was conducted to examine the likelihood of reverse causality, and multivariable MR was performed to adjust the potential confounders. Results: We collected 1,505 host single nucleotide polymorphisms (SNPs) linked to 119 gut microbiota traits and 1,873 host SNPs associated with 81 gut metabolite traits as exposure data. Among these, three gut bacteria indicated an elevated risk of IS, two of ICH, and one of SAH. In contrast, five gut bacteria were associated with a reduced risk of IS, one with ICH, and one with SAH. Our study also demonstrated the potential causal associations between 11 gut microbiota-derived metabolites and CVD. Conclusions: This study provided evidence of the causal relationship between gut microbiota, gut microbiota-derived metabolites, and CVD, thereby offering novel perspectives on gut biomarkers and targeted prevention and treatment for CVD.

High-throughput Sequencing Analysis of Differentially Expressed microRNAs Associated With SREBP1 in Differentiated Thyroid Carcinoma.

BACKGROUND/AIM: MicroRNAs (miRNAs) interact with mRNAs and play important roles in progression and prognosis in multiple cancers. Sterol regulatory element-binding protein 1 (SREBP1) is an important lipid metabolism regulatory gene. The aim of the present study was to analyze the profiles of miRNAs that are associated with SREBP1 expression in differentiated thyroid carcinoma (DTC). MATERIALS AND METHODS: In the present study, a high-throughput small RNA sequencing (miRNA-Seq) method was used to investigate differences in miRNA profiling with versus without interference with SREBP1 expression via small interfering RNA. Real-time qPCR (qRT-PCR) was performed to confirm the results. RESULTS: A total of 1,393 conserved and 84 novel miRNAs were successfully discovered. In two separate batches, a total of 27 differentially expressed miRNAs (11 up-regulated and 16 down-regulated) were observed in BCPAP cells after SREBF1 interference with two distinct siRNA fragments, as compared to the control siRNA treatment. Hsa-miR-941, hsa-miR-27a-5p, hsa-miR-29a-3p, hsa-miR-100-5p, and hsa-miR-21-3p were selected for validation using qRT-PCR. The qRT-PCR results were consistent with the sequencing data. Gene Ontology enrichment showed that the predicted targets of these miRNAs were mainly involved in the regulation of system development, metabolism and protein binding cellular processes, and metabolic processes. Kyoto Encyclopedia of Genes and Genomes pathways analysis showed that the predicted target genes were involved in several signaling pathways, including the Ras, MAPK, insulin, thyroid hormone, and metabolic pathway signaling pathways. CONCLUSION: Differentially expressed miRNAs and their target genes may play an important role in the progression and prognosis of DTC that is associated with SREBP1 expression.

Transcriptome and the gut microbiome analysis of the impacts of Brucella abortus oral infection in BALB/c mice.

Brucellosis is a zoonotic infectious disease caused by Brucella spp, which could cause serious economic losses to animal husbandry and threaten human public health. Ingestion of contaminated animal products is a common way to acquire Brucella infection in humans, while research on effect of oral Brucella infection on host gut microbiota and the gene expression in intestinal tissues is limited. In the present study, 16S rRNA sequencing and RNA sequencing were conducted to explore gut microbiota and expression profiles of mRNAs in the colon of BALB/c mice, which were infected by Brucella abortus 2308. The fecal samples were collected at 7 and 28 days post infection to observe changes in the gut microbiota during Brucella infection. In the alpha diversity analysis, significantly increased Chao 1 index was observed at 28 days after Brucella infection. The Bray-Curtis distancebased principal coordinate analysis indicated that the WT group showed a separation from the Brucella infection groups. In addition, analysis of composition of microbes revealed that Prevotellaceae_NK3B31_group were more abundant in 1 week and 4 week infection groups, while Turicibacter was only more abundant in 4 week infection group. Based on the RNA-seq assay, a total of 45 differentially expressed genes were detected between Brucella abortus infection group and control group. Furthermore, KEGG pathway enrichment analysis showed that protein processing in endoplasmic reticulum, Legionellosis, Spliceosome, Hippo signaling pathway and Influenza A were significantly enriched in response to Brucella abortus infection. Our finding will help to improve the knowledge of the mechanisms underlying Brucella infection and may provide novel targets for future treatment of this pathogen infection.

ClpP protease modulates bacterial growth, stress response, and bacterial virulence in Brucella abortus.

The process of intracellular proteolysis through ATP-dependent proteases is a biologically conserved phenomenon. The stress responses and bacterial virulence of various pathogenic bacteria are associated with the ATP-dependent Clp protease. In this study, a Brucella abortus 2308 strain, ΔclpP, was constructed to characterize the function of ClpP peptidase. The growth of the ΔclpP mutant strain was significantly impaired in the TSB medium. The results showed that the ΔclpP mutant was sensitive to acidic pH stress, oxidative stress, high temperature, detergents, high osmotic environment, and iron deficient environment. Additionally, the deletion of clpP significantly affected Brucella virulence in macrophage and mouse infection models. Integrated transcriptomic and proteomic analyses of the ΔclpP strain showed that 1965 genes were significantly affected at the mRNA and/or protein levels. The RNA-seq analysis indicated that the ΔclpP strain exhibited distinct gene expression patterns related to energy production and conversion, cell wall/membrane/envelope biogenesis, carbohydrate transport, and metabolism. The iTRAQ analysis revealed that the differentially expressed proteins primarily participated in amino acid transport and metabolism, energy production and conversion, and secondary metabolites biosynthesis, transport and catabolism. This study provided insights into the preliminary molecular mechanism between Clp protease to bacterial growth, stress response, and bacterial virulence in Brucella strains.

From methane to value-added bioproducts: microbial metabolism, enzymes, and metabolic engineering.

Methane is abundant in nature, and excessive emissions will cause the greenhouse effect. Methane is also an ideal carbon and energy feedstock for biosynthesis. In the review, the microorganisms, metabolism, and enzymes for methane utilization, and the advances of conversion to value-added bioproducts were summarized. First, the physiological characteristics, classification, and methane oxidation process of methanotrophs were introduced. The metabolic pathways for methane utilization and key intermediate metabolites of native and synthetic methanotrophs were summarized. Second, the enzymatic properties, crystal structures, and catalytic mechanisms of methane-oxidizing and metabolizing enzymes in methanotrophs were described. Third, challenges and prospects in metabolic pathways and enzymatic catalysis for methane utilization and conversion to value-added bioproducts were discussed. Finally, metabolic engineering of microorganisms for methane biooxidation and bioproducts synthesis based on different pathways were summarized. Understanding the metabolism and challenges of microbial methane utilization will provide insights into possible strategies for efficient methane-based synthesis.

Challenging routine: technical difficulties and solutions of endoscopic thyroidectomy via a combined transoral and breast approach - a case-series and learning curve.

OBJECTIVE: Conventional cervical lymph node dissection often leaves large surgical scars, which seriously compromises the postoperative aesthetic effect and can affect the quality of life of patients. In this study, the safety and feasibility of robotic-assisted endoscopic thyroidectomy with central neck dissection (CND) and lateral neck dissection (LND) via a combined transoral and breast approach are discussed in detail. MATERIALS AND METHODS: A retrospective analysis was made of the data of 26 patients with stage cN1b papillary thyroid carcinoma who were admitted to the Thyroid Surgery Department of the Hunan Cancer Hospital from March 2021 to September 2022 and who underwent robotic-assisted endoscopic thyroidectomy with LND via a combined transoral and breast approach. The demographic data, surgical indicators, postoperative data, and the postoperative complication rate of the patients were analyzed, and the learning curve was analyzed by cumulative summation. RESULTS: All the patients underwent endoscopic surgery without any conversion to open surgery. The mean operation time was 313.7±50.3 min and the mean number of total positive/retrieved lymph nodes was 11.2±8.1/36.8±13.7. Two patients developed temporary laryngeal recurrent nerve palsy and three patients developed temporary hypoparathyroidism, all of whom recovered within 3 months postoperatively. No tumor recurrence occurred during follow-ups that ranged from 6 to 24 months. The mean postoperative quality of life (QOL) score was 189.1±118.2, test results ranging from 0 to 1300 with a lower score indicating a higher QOL, and the aesthetic satisfaction score was 4.2±0.7, test scores ranging from 0 to 5 with higher scores indicate higher satisfaction. The turning point of the learning curve was in the 11th case. CONCLUSIONS: The robotic-assisted endoscopic thyroidectomy with CND and LND via a combined transoral and breast approach is safe and feasible, and the improved cosmetic effect is remarkable, which is conducive to improving the postoperative QOL of patients. It provides a new surgical option for patients.

Comparative study of different sewage sludge incineration treatments based on environmental and economic life cycle assessment.

Incineration is one of the most widely used treatments in the field of sewage sludge disposal. However, the choice of sewage sludge incineration process is still controversial. In this study, the comparative life cycle assessment of sewage sludge incineration processes, including the mono-incineration, co-incineration in coal-fired power plants and co-incineration in municipal solid waste (MSW) incineration plants, was carried out from the perspective of environment, carbon footprint and economy. The environmental assessment results show that terrestrial ecotoxicity, freshwater ecotoxicity, marine ecotoxicity, human carcinogenic toxicity and human non-carcinogenic toxicity are the most significant environmental impacts. And the environmental performance of co-incineration in coal-fired power plants is the best. Moreover, the environmental impact is most sensitive to the dehydrant, electricity and fly ash chelating agent. Co-incineration in MSW incineration plants has the lowest carbon emissions, with only 70.50% and 82% of the carbon emissions from mono-incineration and co-incineration in coal-fired power plants, respectively. Furthermore, sewage sludge mono-incineration has the highest disposal costs because of the higher depreciation and solid waste disposal costs. The comprehensive evaluation results reveal that the optimization should focus on the selection of dehydrant and fly ash chelating agent, as well as the improvement of the equipment efficiency.

Sequential extraction, structural characterization, and antioxidant activity of polysaccharides from Dendrocalamus brandisii bamboo shoot shell.

Polysaccharides including water-soluble fraction (W), 1,2-Cyclohexanediaminetetraacetic acid (CDTA)-soluble fraction (CA), sodium carbonate (Na2CO3)-soluble fraction (SC), 1 M potassium hydroxide (KOH)-soluble fractions (PH1), and 4 M KOH-soluble fraction (PH4) were successively extracted from Dendrocalamus brandisii bamboo shoot shells using water, CDTA, Na2CO3, and KOH solution. The analytical methods were employed to initially identify the structural characteristics of the five polysaccharide fractions, and their antioxidant capacities in vitro were determined. According to the data, the average molecular weight of the five polysaccharide fractions was between 4 816 and 993 935 Da. In all four types (CA, SC, PH1, and PH4), xylose was the most abundant monosaccharide, especially in PH1 and PH4. Both PH1 and PH4 were found to contain 1,4-ß-d-Xylp as their main chain, as determined by nuclear magnetic resonance (NMR) spectroscopy. Additional research into CA and SC's antioxidant potential is required since they both showed potent in vitro antioxidant activities.

Comprehensive evaluation of sewage sludge anaerobic digestion process with different digestate treatments.

Anaerobic digestion is one of the most promising methods for reducing sewage sludge and recovering energy. In the present study, a comparative life cycle assessment (LCA) of sewage sludge anaerobic digestion processes with different digestate treatments, including mesophilic anaerobic digestion with digestate landfilling (CAD-1) and digestate incineration (CAD-2), thermophilic anaerobic digestion combined with thermal hydrolysis pre-treatment with digestate land use (THPAD-1), and digestate incineration (THPAD-2), was performed to evaluate their environmental, resource, economic, and comprehensive performances using the SimaPro software. Environmental impact analysis revealed marine ecotoxicity, freshwater ecotoxicity, and human carcinogenic toxicity as the most obvious impacts, resulting in the most significant damage to human health. Resource analysis indicated that anaerobic digestion combined with cogeneration and digestate incineration is advantageous to high energy recovery, but digestate incineration is disadvantageous to economic performance because of increased investment costs. Comparison of the four processes revealed that THPAD-2 results in the largest environmental damage, whereas CAD-1 has the smallest load. Meanwhile, THPAD-2 and THPAD-1 exhibit the best resource performance and net economic benefit, respectively. The comprehensive evaluation indices revealed that THPAD-1 and CAD-2 show better comprehensive performance. And the deep drying incineration process exhibited better comprehensive performance than sewage sludge anaerobic digestion processes.

Co-conversion of lignocellulose-derived glucose, xylose, and aromatics to polyhydroxybutyrate by metabolically engineered Cupriavidus necator.

Utilization of all major components of lignocellulose is essential for biomass biorefining. Glucose, xylose, and lignin-derived aromatics can be generated from cellulose, hemicellulose, and lignin of lignocellulose degradation through pretreatment and hydrolysis. In present work, Cupriavidus necator H16 was engineered to utilize glucose, xylose, p-coumaric acid, and ferulic acid simultaneously by multi-step genetic engineering. Firstly, genetic modification and adaptive laboratory evolution were performed to promote glucose transmembrane transport and metabolism. Xylose metabolism was then engineered by integrating genes xylAB (xylose isomerase and xylulokinase) and xylE (proton-coupled symporter) in the locus of ldh (lactate dehydrogenase) and ackA (acetate kinase) on the genome, respectively. Thirdly, p-coumaric acid and ferulic acid metabolism was achieved by constructing an exogenous CoA-dependent non-ß-oxidation pathway. Using corn stover hydrolysates as carbon sources, the resulting engineered strain Reh06 simultaneously converted all components of glucose, xylose, p-coumaric acid, and ferulic acid to produce 11.51 g/L polyhydroxybutyrate.

Applications and Challenges of Supercritical Foaming Technology.

With economic development, environmental problems are becoming more and more prominent, and achieving green chemistry is an urgent task nowadays, which creates an opportunity for the development of supercritical foaming technology. The foaming agents used in supercritical foaming technology are usually supercritical carbon dioxide (ScCO2) and supercritical nitrogen (ScN2), both of which are used without environmental burden. This technology can reduce the environmental impact of polymer foam production. Although supercritical foaming technology is already in production in some fields, it has not been applied on a large scale. Here, we present a detailed analysis of the types of foaming agents currently used in supercritical foaming technology and their applications in various fields, summarizing the technological improvements that have been made to the technology. However, we have found that today's supercritical technologies still need to address some additional challenges to achieve large-scale production.

The Pseudogene BMEA_B0173 Deficiency in Brucella melitensis Contributes to M-epitope Formation and Potentiates Virulence in a Mice Infection Model.

It is widely acknowledged that pseudogenes play important roles in bacterial diversification and evolution and participate in gene regulation and RNA interference (RNAi). However, the function of most pseudogenes in Brucella spp remains poorly understood, warranting further studies.To comprehensively analyze the function of the pseudogenes BMEA_B0173 in Brucella melitensis strain 63/9, a BMEA_B0173 in-frame deleted mutant strain was constructed. Then, the phenotypes of the mutant strain, such as growth characteristics and bacterial virulence, were assessed in mice infection models. Finally, iTRAQ analysis was performed to investigate the gene expression profile affected by the pseudogenes BMEA_B0173. In this study, we found that BMEA_B0173 deletion exhibited increased agglutination with M monospecific sera. In a mouse model of chronic infection, the BMEA_B0173 deletion strain displayed increased colonization in the spleen compared to the wild-type pathogen. The iTRAQ assay revealed that 252 proteins were differentially expressed between the BMEA_B0173 deletion and the wild-type strains. In addition, deletion of BMEA_B0173 significantly increased the expression of proteins involved in the denitrification pathway, iron metabolism, and several transcriptional regulators, which might cause increased virulence of the mutant strain. In conclusion, this study preliminary uncovered the function of the pseudogene BMEA_B0173 in Brucella melitensis 63/9 and provided novel insights for studying the pathogenesis of Brucella strains.

Pal Affects the Proliferation in Macrophages and Virulence of Brucella, and as Mucosal Adjuvants, Provides an Effective Protection to Mice Against Salmonella Enteritidis.

The purpose of this study was to elucidate the roles of peptidoglycan-associated lipoprotein (Pal protein) in the proliferation of Brucella in macrophage and bacterial virulence, and to evaluate the immune effect of Pal protein to Salmonella enteritidis. Murine macrophage-like cell line Raw264.7 was stimulated by recombinant Pal protein, and the expression of TNF-α and IFN-γ were up-regulated, but not it of IL-1ß and IL-6. The macrophages infection and in vitro simulated stress assays showed that deletion of pal gene reduced the proliferation of Brucella in macrophages, the survival in acidic, oxidative and polymyxin B-contained environment. The mice infection assay showed that mice challenged with the pal mutant strain were found to have more severe splenomegaly, but less bacterial load. After oral immunization of mice, Pal protein induced a higher titer of mucosal and humoral antibody (IgA and IgG) against heat-killed Salmonella enteritidis, and a stronger Th1 cellular immune response. The challengte experiments showed Pal protein elevated the survival rate and reduced the bacterial load of spleens in immunized mice. In conclusion, our results revealed the important roles of pal gene in Brucella virulence, and Pal protein was a potentially valuable adjuvant against mucosal pathogens, such as Salmonella enteritidis.

Combination effect between gut microbiota and traditional potentially modifiable risk factors for first-ever ischemic stroke in Tujia, Miao and Han populations in China.

China has had explosive growth in ischemic stroke (IS) burden with significant ethnic and geographic disparities. The aim of this study was to explore the possible combination effect between gut microbiota and traditional potentially modifiable risk factors for IS among two ethnic minorities (Tujia and Miao) and the Han population. Herein, we first used the 16 S rRNA sequencing to compare the gut microbial compositions of 82 patients with first-ever IS vs. 82 normal controls (NCs) among Han, Tujia, and Miao people between 1 May 2018 and 30 April 2019, from Xiangxi Tujia and Miao Autonomous Prefecture in China. An additive model was used to study the interaction between traditional risk factors and gut microbiota with R software. Linear discriminant analysis (LDA) and LDA effect size (LEfSe) results showed that the identified key gut microbiota's taxonomic composition varied in different ethnicity between the IS patients and NCs. Furthermore, families Lactobacillaceae, Enterococcaceae, Streptococcaceae, and Enterobacteriaceae were found to be positively correlated with high-risk factors and negatively correlated with preventive factors in the IS patients, but families Ruminococcaceae and Lachnospiraceae were just the opposite in the NCs. There were additive interactions between traditional risk factors (systolic blood pressure, diastolic blood pressure, and high-sensitive C-reactive protein) and family Enterococcaceae for first-ever IS with the attributable proportion due to the interaction was 0.74, 0.71, and 0.85, respectively; and the synergy index was 4.45, 3.78, and 7.01, respectively. This preliminary but promising study showed that the gut microbiota disturbances may potentially interact to IS with different ethnic host's traditional risk factors.

Evaluation of the in vitro antioxidant and antitumor activity of extracts from Camellia fascicularis leaves.

Camellia fascicularis is a unique plant rich in bioactive components. However, the isolation of the active substances in C. fascicularis leaves via sequential extraction with solvents of different polarity and the determination of their antioxidant and antitumor activities have not been reported. In this study, the total methanol extract of C. fascicularis leaves was sequentially extracted with different polar solvents, and the corresponding petroleum ether extract (PEE), ethyl acetate extract (EAE), and water extract (WE) were analyzed for their contents in active substances such as flavonoids, polyphenols, polysaccharides, and saponins. The antioxidant ability of the polar extracts was investigated by determining their reducing power and the radical scavenging rate on 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and hydroxyl radicals, and CCK-8 and Annexin-FITC/propidium iodide staining assays were conducted to investigate their inhibitory effects on HCCLM6 and HGC27 tumor cells. The results showed that PEE had a high saponin content of 197.35 ± 16.21 mg OAE/g, while EAE and WE exhibited a relatively higher polysaccharide content of 254.37 ± 1.99 and 373.27 ± 8.67 mg GE/g, respectively. The EAE demonstrated the greatest reducing power and the strongest clearing abilities on ABTS and DPPH radicals with respective EC50 values of 343.45 ± 20.12 and 14.07 ± 0.06 µg/ml. Moreover, the antitumor ability of the different polar extracts was dose-dependent, with WE showing the most potent inhibitory ability against HCCLM6 and HGC27 cells.