Protective Effect of <i>Eurotium cristatum</i> Fermented Loose Dark Tea and <i>Eurotium cristatum</i> Particle on MAPK and PXR/AhR Signaling Pathways Induced by Electronic Cigarette Exposure in Mice

Electronic-cigarette smoke (eCS) has been shown to cause a degree of oxidative stress and inflammatory damage in lung tissue. The aim of this study was to evaluate the repair mechanism of Eurotium cristatum fermented loose dark tea (ECT) and Eurotium cristatum particle metabolites (ECP) sifted from ECT after eCS-induced injury in mice. Sixty C57BL/6 mice were randomly divided into a blank control group, an eCS model group, an eCS + 600 mg/kg ECP treatment group, an eCS + 600 mg/kg ECT treatment group, an eCS + 600 mg/kg ECP prevention group, and an eCS + 600 mg/kg ECT prevention group. The results show that ECP and ECT significantly reduced the eCS-induced oxidative stress and inflammation and improved histopathological changes in the lungs in mice with eCS-induced liver injury. Western blot analysis further revealed that ECP and ECT significantly inhibited the eCS-induced upregulation of the phosphorylation levels of the extracellular Regulated protein Kinases (ERK), c-Jun N-terminal kinase (JNK) and p38mitogen-activated protein kinases (p38MAPK) proteins, and significantly increased the eCS-induced downregulation of the expression levels of the pregnane X receptor (PXR) and aryl hydrocarbon receptor (AhR) proteins. Conclusively, these findings show that ECP and ECT have a significant repairing effect on the damage caused by eCS exposure through the MAPK and PXR/AhR signaling pathways; ECT has a better effect on preventing eCS-induced injury and is suitable as a daily healthcare drink; ECP has a better therapeutic effect after eCS-induced injury, and might be a potential therapeutic candidate for the treatment of eCS-induced injury

Effect of Fermentation by Eurotium cristatum LJSC.2001 on the Fermentation Quality of Raw Dark Tea Made from Different Varieties

The current study was performed in order to explore the effect of Eurotium cristatum LJSC.2001 on the fermentation quality of raw dark tea. Golden flower loose tea was produced by using E. cristatum LJSC.2001 to ferment raw dark tea from the cultivars Zhuyeqi, Yuntaidaye and Taoyuandaye provided by representative manufacturers (Yuntaishan, Yunshang, Baishaxi and Junhe) in Anhua, Hunan province, and comparative analysis of their taste and aroma quality was carried out by sensory evaluation, basic physicochemical analysis, analysis of catechin components by high performance liquid chromatography (HPLC), and headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS). Results showed that after fermentation by E. cristatum LJSC.2001, the dry tea samples were darker in color and fully covered with golden flower; their infusions were darker in color, and had a strong fungal aroma and a mellow and full taste, and the brewed tea was soft. Raw dark tea from Baishaxi had a stronger fungal aroma after fermentation, while retaining a distinct pine smoke-like aroma. The contents of taste compounds including tea polyphenols, free amino acids, soluble carbohydrates, flavonoids, total catechins, myricetin, quercetin, and kaempferol showed an overall decreasing trend. The content of catechins in Yuntaidaye raw dark tea from Yuntaishan and Yunshang declined most. The relative contents of esters and aldehydes increased after fermentation. There were differences in the content of volatile components among the different fermented tea samples, methyl salicylate and (E,E)-2,4-heptadienal being the major ones. Multivariate statistical analysis showed significant differences in volatile composition between golden flower loose tea and raw black tea. There were 19 characteristic volatile components identified including methyl salicylate, (E,E)-2,4-heptadienal, (E)-linalool 3,7-oxide, (E)-furan oxidized linaloa, acetophenone, (E)-2-,(Z)-6-nonadienal, (E)-2-hexenal, (E,E)-3,5-octadien-2-one and n-tridecane. The contents of the nine volatile components increased significantly after fermentation, and they all contributed to the fungal aroma of golden flower loose tea. It can be seen that fermentation by E. cristatum LJSC.2001 can impart a more mellow and fuller taste and a more prominent fungal aroma to raw black tea

Risk factors associated with the prevalence of thyroid nodules in adults in Northeast China: a cross-sectional population-based study

Objectives This study examined the association between anthropometric measurements, lifestyle factors and the prevalence of thyroid nodules among adults in Northeast China.Design We employed a cross-sectional approach involving a questionnaire survey, which focused on participants’ living habits, and a physical examination that included anthropometry and ultrasound imaging.Setting The data were procured during multiple trips by medical teams from the first hospital of China Medical University to towns in Northeast China.Participants Of the 1092 participants, 489 did not have thyroid nodules (mean age: 54.02±11.49 years; 297 females (60.7%)), 99 had single thyroid nodules (mean age: 58.19±10.77 years; 59 females (59.6%)) and 504 had multiple thyroid nodules (mean age: 60.05±10.68 years; 394 females (78.2%)). Inclusion criteria mandated participants be over 20 years old without other medical conditions. We excluded individuals who had undergone surgical resection for thyroid nodules.Results The prevalence of thyroid nodules was significantly associated with being female (OR 2.569, 95% CI 1.937 to 3.405, p&lt;0.001) and increased age (OR 1.054, 95% CI 1.041 to 1.066, p&lt;0.001). This association was more pronounced in those with multiple thyroid nodules. For males under 60, non-smoking was inversely correlated with the prevalence of multiple thyroid nodules (OR 0.321, 95%CI 0.149 to 0.69, p&lt;0.05). For females under 60, diastolic blood pressure (DBP) was significantly linked with the prevalence of thyroid nodules (OR 0.978, 95% CI 2.614 to 2.705, p&lt;0.05).Conclusions Besides gender and age, the prevalence of thyroid nodules in Northeast China correlates with smoking habits and DBP

Palladium-Catalyzed Oxidative C–N Bond Coupling Involving a Solvent-Controlled Regioselective Bromination Process

Stereoselective palladium-catalyzed oxidative C–N bond coupling reactions between aromatic amines and alkenes involving a solvent-controlled regioselective bromination process under 1 atm of oxygen atmosphere are disclosed, providing easy access to two different brominated enamines. The addition of hydrogen peroxide (30% aq) as a co-oxidant in the system is crucial for the dehydrogenative aminohalogenation under molecular oxygen (1 atm), and in such a case, the C–N bond coupling/electrophilic bromination reaction cascade is proposed. Furthermore, the different reaction media leads to a switched regioselectivity of the process

The WRKY Transcription Factor CsWRKY70 Regulates EGCG Biosynthesis by Affecting <i>CsLAR</i> and <i>CsUGT84A</i> Expressions in Tea Leaves (<i>Camellia sinensis</i>)

Epigallocatechin gallate (EGCG) is an important contributor to bitterness and astringency in summer tea leaves; however, the transcriptional regulatory mechanisms of EGCG biosynthesis remain unclear. In this study, EGCG content was significantly decreased after foliar spraying with nano-Se fertilizers in tea leaves. A WRKY transcription factor (TF), CsWRKY70, was found to be positively related to EGCG content. The open reading frame of CsWRKY70 was 891 bp encoding 296 amino acids. CsWRKY70 is localized to the nucleus and has transcriptional activation activity. The electrophoretic mobility shift assay indicated that CsWRKY70 can directly bind to the promoters of CsLAR and CsUGT84A containing W-box (5′-C/TTGACT/C-3′) sequences. Dual-luciferase reporter experiment verified that CsWRKY70 activated CsLAR and CsUGT84A expressions in tobacco leaves. In summary, these results demonstrated that CsWRKY70 may reduce EGCG biosynthesis by inhibiting the CsLAR and CsUGT84A expressions under nano-Se treatment. Our findings provide new insight into the regulatory mechanism of WRKY TFs involved in catechin biosynthesis and offer a theoretical basis for breeding low or high EGCG content tea cultivars

Identification, Molecular Characteristic, and Expression Analysis of PIFs Related to Chlorophyll Metabolism in Tea Plant (Camellia sinensis)

The phytochrome-interacting factors (PIFs) proteins belong to the subfamily of basic helix–loop–helix (bHLH) transcription factors and play important roles in chloroplast development and chlorophyll biosynthesis. Currently, knowledge about the PIF gene family in Camellia sinensis remains very limited. In this study, seven PIF members were identified in the C. sinensis genome and named based on homology with AtPIF genes in Arabidopsis thaliana. All C. sinensis PIF (CsPIF) proteins have both the conserved active PHYB binding (APB) and bHLH domains. Phylogenetic analysis revealed that CsPIFs were clustered into four groups—PIF1, PIF3, PIF7, and PIF8—and most CsPIFs were clustered in pairs with their corresponding orthologs in Populus tremula. CsPIF members in the same group tended to display uniform or similar exon–intron distribution patterns and motif compositions. CsPIF genes were differentially expressed in C. sinensis with various leaf colors and strongly correlated with the expression of genes involved in the chlorophyll metabolism pathway. Promoter analysis of structural genes related to chlorophyll metabolism found DNA-binding sites of PIFs were abundant in the promoter regions. Protein–protein interaction networks of CsPIFs demonstrated a close association with phytochrome, PIF4, HY5, TOC1, COP1, and PTAC12 proteins. Additionally, subcellular localization and transcriptional activity analysis suggested that CsPIF3b was nuclear localized protein and possessed transcriptional activity. We also found that CsPIF3b could activate the transcription of CsHEMA and CsPOR in Nicotiana benthamiana leaves. This work provides comprehensive research of CsPIFs and would be helpful to further promote the regulation mechanism of PIF on chlorophyll metabolism in C. sinensis

A Comparative Proteomic Analysis of the Buds and the Young Expanding Leaves of the Tea Plant (Camellia sinensis L.)

Tea (Camellia sinensis L.) is a perennial woody plant that is widely cultivated to produce a popular non-alcoholic beverage; this beverage has received much attention due to its pleasant flavor and bioactive ingredients, particularly several important secondary metabolites. Due to the significant changes in the metabolite contents of the buds and the young expanding leaves of tea plants, high-performance liquid chromatography (HPLC) analysis and isobaric tags for relative and absolute quantitation (iTRAQ) analysis were performed. A total of 233 differentially expressed proteins were identified. Among these, 116 proteins were up-regulated and 117 proteins were down-regulated in the young expanding leaves compared with the buds. A large array of diverse functions was revealed, including roles in energy and carbohydrate metabolism, secondary metabolite metabolism, nucleic acid and protein metabolism, and photosynthesis- and defense-related processes. These results suggest that polyphenol biosynthesis- and photosynthesis-related proteins regulate the secondary metabolite content of tea plants. The energy and antioxidant metabolism-related proteins may promote tea leaf development. However, reverse transcription quantitative real-time PCR (RT-qPCR) showed that the protein expression levels were not well correlated with the gene expression levels. These findings improve our understanding of the molecular mechanism of the changes in the metabolite content of the buds and the young expanding leaves of tea plants