Amino acids and flavonoids analysis reveals quality constituents difference among different albino tea resources.

Albino tea has attracted increased attention due to its unique flavor. To reveal the difference in key metabolites constituting the important quality of different tea resources, amino acids and flavonoids profiles in three albino resources with different degrees of albinism and one normal green variety were comprehensively investigated. K-means analysis revealed 35 amino acids were significantly enriched in 'Jibai', while 3 and 2 were specifically accumulated in 'Huangjinya' and 'Anjibaicha', respectively. Based on OPLS-DA models, 40, 31 and 45 significantly differential flavonoids were determined in 'Huangjinya', 'Anjibaicha' and 'Jibai' compared to 'Fudingdabaicha', and most were down-regulated. Among them, 10, 5 and 13 differential flavonoids were exclusively found in 'Huangjinya', 'Anjibaicha' and 'Jibai', respectively, which may contribute to unique quality for different resources. The differential flavonoids and amino acids involved in their metabolic pathways were obviously different among four resources, resulting in the difference in tea quality and flavor.

Effect of blister blight disease caused by Exobasidium on tea quality.

Blister blight, as one of the most threatening and damaging disease worldwide, mainly infects young organs and tissues seriously affecting tea growth and quality. In this study, the spread of pathogen on tea leaves were examined by toluidine blue staining, scanning electron microscope and transmission electron microscope analysis. The composition and abundance of fungal community on leaf tissues were firstly analyzed. Sensory evaluation and metabolites analysis indicated that diseased tea leaves had strong sweet taste and soluble sugars contributed significantly to the taste, while metabolites showing bitter and astringent taste (caffeine, catechins) were significantly decreased. According to the biological functions of differential metabolites, sugars including 7 monosaccharides (d-xylose, d-arabinose, d-mannose, d-glucuronic acid, glucose, d-galactose and d-fructose), 2 disaccharide (sucrose and maltose) and 1 trisaccharide (raffinose) were the main differential sugars increased in content (>2 fold change), which was of great significance to sweet taste of diseased tea.

Metabolites profiling reveals the dynamic changes of non-volatiles in Pu-erh during Ganpu tea processing.

Ganpu is an unique tea product made by Pu-erh tea and citrus peel. In this study, the non-volatiles changes of Pu-erh during Ganpu tea processing were fully analyzed by UPLC-ESI-MS/MS. Total 276 significantly differential metabolites in Pu-erh during Ganpu processing were detected (P < 0.05, VIP > 1), and their change trend were clustered into 8 subclasses by K-means analysis. Metabolites of Pu-erh present at various processes were revealed. 72 differential metabolites (P < 0.05, VIP > 1 and fold change ≥2 or ≤0.5) between any two stages were identified and fixation was the key step with 61 differential metabolites. 39 flavonoids and 2 lignans and coumarins were significantly decreased after fixation, while 5 terpenoids, 3 amino acids, 1 organic acids, 2 nucleotides and derivatives and newly detected jasminoside A (Log2FC = 9.90), picrocrocin (Log2FC = 9.90) and nomilinic acid (Log2FC = 7.56) were significantly increased. The results provided valuable information about the effect of Ganpu processing on dynamic changes of non-volatiles in Pu-erh.

Sensory-directed isolation and identification of an intense salicin-like bitter compound in infected teas with bird's eye spot disease.

Teas infected with bird's eye spot disease generally exhibited a lingering and long-lasting, salicin-like bitter taste, which was unpalatable to consumers. Sensory-directed isolation processes have been performed in this study to investigate the salicin-like bitter compounds in infected teas. Results showed that infected teas were extracted using a 70% methanol aqueous solution to produce methanol extract, which was then further separated by sequential solvent extraction (SSE) to obtain dichloromethane extract, which contained the salicin-like bitter compounds. The dichloromethane extract was then isolated by flash chromatography to produce two salicin-like bitter fractions, eluted using 60% and 65% methanol aqueous solution. Finally, these two salicin-like bitter fractions were analyzed by RP-HPLC using 60-68% and 70-75% methanol aqueous solution, respectively, affording the location of the salicin-like bitter compounds in RP-HPLC chromatograms. Moreover, a new ursane-type triterpenoid, camellisin A methyl ester, was identified from infected teas. This study has provided preliminary isolation methods of salicin-like bitter compounds from the infected teas, which were essential to designing targeted debittering strategies for infected teas and improving the quality of the finished tea and the effective utilization of fresh tea leaves.

Metabolites and metagenomic analysis reveals the quality of Pu-erh "tea head".

Tea head, a derivative product of Pu-erh tea, are tight tea lumps formed during pile-fermentation. The aim of this study was to reveal the differences of quality-related metabolites and microbial communities between ripened Pu-erh tea (PE-21) and tea heads (CT-21). Compared with PE-21, CT-21 showed a more mellow and smooth taste with slight bitterness and astringency, and can withstand multiple infusions. Metabolites analysis indicated CT-21 had more abundant water-soluble substances (47.39%) and showed significant differences with PE-21 in the main compositions of amino acids, catechins and saccharides which contributed to the viscosity of tea liquor, mellow taste and the tight tea lumps formation. Microbial communities and COG annotation analysis revealed CT-21 had lower abundance of Bacteria (84.05%), and higher abundance of Eukaryota (15.10%), carbohydrate transport and metabolism (8.28%) and glycoside hydrolases (37.36%) compared with PE-21. The different microbial communities may cause metabolites changes, forming distinct flavor of Pu-erh.

Metabolomics Analysis Reveals the Effect of Two Alpine Foliar Diseases on the Non-Volatile and Volatile Metabolites of Tea.

Blister blight and small leaf spots are important alpine diseases that mainly attack tender tea leaves, affecting tea quality. However, there is limited information on the effect of these diseases on tea's non-volatile and volatile metabolites. Metabolomic analysis based on UHPLC-Q-TOF/MS, HPLC and GC/MS was used to reveal the characteristic chemical profiles of tea leaves infected with blister blight (BB) and small leaf spots (SS). Flavonoids and monolignols were non-volatile metabolites that were enriched and significantly changed. Six main monolignols involved in phenylpropanoid biosynthesis were significantly induced in infected tea leaves. The accumulation of catechins, (-)-epigallocatechin gallate, (-)-epicatechin gallate, caffeine, amino acids and theanine were significantly decreased in both diseased tea leaves, while soluble sugar, (-)-epigallocatechin and phenol-ammonia were obviously increased. Among them, the amounts of sweet and umami-related soluble sugar, sucrose, amino acids and theanine were much higher in BB, while bitter and astringent taste-related catechins and derivatives were much higher in SS. Volatiles analysis showed that volatiles content in SS and BB was significantly decreased, and styrene was significantly induced in blister blight-infected tea leaves. The results indicate that the type and amount of volatiles were highly and differentially influenced by infection with the two alpine diseases.

Characterization of triterpenoids as possible bitter-tasting compounds in teas infected with bird's eye spot disease.

Tea infected with bird's eye spot disease generally imparts a long-lasting bitter taste, which is unacceptable to most consumers. This study has comprehensively evaluated the taste profiles of infected and healthy teas and investigated their known bitter compounds previously reported in tea. Quantification analyses and calculation of dose-over-threshold (DoT) factors revealed that no obvious difference was visualized in catechins, caffeine, bitter amino acids, and flavonols and their glycosides between infected and healthy tea samples, which was also verified by principal component analysis (PCA) and hierarchical cluster analysis (HCA). Therefore, these known bitter compounds have been ruled out as critical contributors to the long-lasting bitterness of infected teas. Furthermore, Gel permeation chromatography, sensory analysis, and UPLC-Q-TOF-MS were employed and identified 13 substances from the target bitter fractions, including caffeine, ten triterpenoids, and two oxylipins. The higher triterpenoid levels were supposed to be the reason causing the long-lasting bitterness. This study has provided a research direction for the molecular basis of the long-lasting bitterness of infected tea leaves with bird's eye spot disease.

Characteristics of Leaf Spot Disease Caused by Didymella Species and the Influence of Infection on Tea Quality.

Leaf spots are the most damaging and common foliar diseases of tea and are caused by several species of fungi. During 2018 to 2020, leaf spot diseases showing different symptoms (large and small spots) were observed in commercial tea plantations in Guizhou and Sichuan provinces of China. The pathogen causing the two different sized leaf spots was identified as the same species (Didymella segeticola) based on morphological characteristics, pathogenicity, and multilocus phylogenetic analysis using the combined ITS, TUB, LSU, and RPB2 gene regions. Microbial diversity analysis of lesion tissues from small spots on naturally infected tea leaves further confirmed Didymella to be present as the main pathogen. Results of sensory evaluation and quality-related metabolite analysis of tea shoots infected with the small leaf spot symptom indicated that D. segeticola negatively affected the quality and flavor of tea by changing the composition and content of caffeine, catechins, and amino acids. In addition, the significantly reduced amino acid derivatives in tea are confirmed to be positively associated with the enhanced bitter taste. The results improve our understanding of the pathogenicity of Didymella species and the influence of Didymella on the host plant, Camellia sinensis.

Non-volatile metabolites profiling analysis reveals the tea flavor of "Zijuan" in different tea plantations.

Characteristic metabolites including tea polyphenols, amino acids, catechins, caffeine, sugars and anthocyanins were fully analyzed by high performance liquid chromatography (HPLC), gas chromatography tandem mass spectrometry (GC-MS) and ultra-high performance liquid chromatography (UHPLC)-ESI-tandem mass spectrometry (MS/MS), and showed significant differences among Zijuan tea from different plantations in Yunnan province (YN-ZJ), Qijiang (QJ-ZJ) and Ersheng (ES-ZJ) district, China, indicating that Zijuan is significantly influenced by growth conditions. Monosaccharides were the most abundant soluble sugars in YN-ZJ and ES-ZJ, while disaccharides was abundant in QJ-ZJ. d-galactose, d-mannose, d-sorbitol, inositol, d-glucose, d-galacturonic acid and raffinose involved in galactose metabolism were significantly changed (P < 0.05). Delphinidin, cyanidin, pelargonidin and their glycoside derivatives were the major anthocyanins, and showed significant differences among Zijuan samples. Flavonoids and procyanidins abundant in Zijuan provided more substrates for anthocyanins accumulation. This study presented comprehensive chemical profiling and characterized metabolites of Zijuan in different tea plantations.

A New Colletotrichum Species Associated with Brown Blight Disease on Camellia sinensis.

Brown blight, as the most damaging and common foliar disease of the tea plant (Camellia sinensis) in China, has been recently reported to be caused by different species of the genus Colletotrichum. During the years 2016 to 2017, tea plants in commercial tea cultivation areas of Chongqing City that reported significant incidences of brown blight disease were investigated and then analyzed using both morphological characteristics and multilocus phylogenetic analysis. The results showed that at least five species of Colletotrichum were identified, including four well-known species (Colletotrichum gloeosporioides, C. camelliae, C. fioriniae, and C. karstii) and one novel species (C. chongqingense), indicating that there is remarkable species diversity in Colletotrichum spp. present as pathogens. Results of pathogenicity analyses confirmed that C. chongqingense was the causal agent of brown blight and different isolates differed in virulence. C. chongqingense, as a novel pathogen, has never been reported as being associated with brown blight disease in tea plants or anthracnose in other host plants anywhere in the world. Knowledge of the Colletotrichum populations will facilitate further studies addressing the relationships between Colletotrichum spp. and their host plant Camellia sinensis.

Guidelines for the diagnosis and treatment of osteoarthritis in China (2019 edition).

Osteoarthritis (OA) is a degenerative disease of middle-aged and elderly people, contributed a higher burden of disease in China and the world. In 2017, under the support of the Rheumatology and Immunology Expert Committee of the Cross-Strait Medical and Health Exchange Association. The objective was to develop an evidence-based diagnosis and treatment guideline for OA in China based on emerging new evidence. The guideline was registered at International Practice Guidelines Registry Platform (IPGRP-2018CN028). The grading of recommendations assessment, development and evaluation (GRADE) approach was used to rate the quality of evidence and the strength of recommendations, and the RIGHT (Reporting Items for Practice Guidelines in Healthcare) checklist was followed to report the guideline. The guideline provides recommendations for the OA diagnosis, disease risks monitoring and evaluate, treatment purpose and physical, medical and surgical interventions. This guideline is intended to serve as a tool for Chinese clinicians for the best decisions-making on diagnosis and treatment of OA.

Development and formulation of the classification criteria for osteoarthritis.

BACKGROUND: The classification criteria of osteoarthritis (OA) is lack of the support of relevant research evidence and there is no standardized protocol for detailed steps of the development or clinical verification of classification criteria has yet been established. This study aims to describe the development process of the Categorization of Osteoarthritis CHecklist (COACH), which is designed to choose the precise treatment option for patients with OA. METHODS: A multidisciplinary panel was established to gather opinions. We conducted questionnaire survey and literature review to generate and COACH Panel members were invited to review the drafted classification criteria and optimize classification criteria. The final list of items was discussed and reached the agreement by the core group of the panel. RESULTS: Thirty-six experts participated in COACH Panel including rheumatologist (80.6%; 29/36), orthopedist (13.9%; 5/36), methodologist (2.8%; 1/36) and rehabilitation physician (2.8%; 1/36) for classification factors generating and optimizing. The main body of the final classification criteria consists of six types of OA pathogenesis, eight types of medical findings (which can be grouped into two categories), and six types of the location. The final criteria include load-based type, structure-based type, inflammation-based type, metabolic-based type, systemic factor based type and mixed type. CONCLUSIONS: COACH can better help clinicians quickly classify OA patients and help to choose the best treatment option from the aspects of types, findings and locations. What's more, the classification criteria are also helpful to promote the basic medical research and targeted prevention of OA.

Metabolism of Gallic Acid and Its Distributions in Tea (Camellia sinensis) Plants at the Tissue and Subcellular Levels.

In tea (Camellia sinensis) plants, polyphenols are the representative metabolites and play important roles during their growth. Among tea polyphenols, catechins are extensively studied, while very little attention has been paid to other polyphenols such as gallic acid (GA) that occur in tea leaves with relatively high content. In this study, GA was able to be transformed into methyl gallate (MG), suggesting that GA is not only a precursor of catechins, but also can be transformed into other metabolites in tea plants. GA content in tea leaves was higher than MG content-regardless of the cultivar, plucking month or leaf position. These two metabolites occurred with higher amounts in tender leaves. Using nonaqueous fractionation techniques, it was found that GA and MG were abundantly accumulated in peroxisome. In addition, GA and MG were found to have strong antifungal activity against two main tea plant diseases, Colletotrichum camelliae and Pseudopestalotiopsis camelliae-sinensis. The information will advance our understanding on formation and biologic functions of polyphenols in tea plants and also provide a good reference for studying in vivo occurrence of specialized metabolites in economic plants.

Erratum to development and formulation of the classification criteria for osteoarthritis.

[This corrects the article DOI: 10.21037/atm-20-4673.].

Leukocytapheresis Therapy for Rheumatoid Arthritis: Results Compared with Control Trial.

CONTEXT: Rheumatoid arthritis (RA) is a chronic multisystem autoimmune disease, mainly characterized by synovitis and with symmetrical joint involvement. LCAP therapy for RA patients has been shown to be safe and efficacious in some developed countries for over a decade. OBJECTIVE: The study intended to evaluate the efficacy and safety of leukocytopheresis (LCAP) for treatment of rheumatoid arthritis (RA) and to study the influence of treatment on the levels of various serum cytokines. DESIGN: The study was a nonblinded, nonrandomized, controlled trial. SETTING: The study took place in the Department of Rheumatology and Immunology at Beijing Hospital at the National Center of Gerontology in Beijing, China. PARTICIPANTS: Participants were 51 patients with RA at the hospital with a 28-joint disease activity score (DAS28) exceeding the 3.20 needed to fulfill the classification criteria of the American College of Rheumatology (ACR). INTERVENTION: Participants were divided into 2 groups. One group (intervention group) received LCAP therapy (n = 20), while the control group (n = 31) received disease-modifying antirheumatic drugs (DMARDs). Patients receiving the LCAP therapy were treated using a Cellsorba column every 5 days for a total of 5 treatments. OUTCOME MEASURES: Clinical assessment of participants' symptoms included: (1) a tender-joint count, (2) a swollen-joint count, (3) erythrocyte sedimentation rates (ESR), (4) C-reactive protein levels (CRP), (5) a visual analog scale (VAS) for pain, (6) the DAS28 C-reactive protein (DAS28-CRP) scores, and the Health Assessment Questionnaire Disability Index (HAQ-DI). The study also evaluated participants' scores for the American College of Rheumatology (ACR) Core Data Set. Serum collected before and after therapy from both groups was analyzed for the levels of bradykinin, serotonin, heat shock protein 70, human CXC-chemokine ligand 16 (CXCL16), prostaglandin E2, and macrophage inflammation protein 1α. RESULTS: At week 4 for participants receiving the LCAP therapy, ACR20, ACR50, and ACR70 were observed in 55%, 30%, and 20% of patients, respectively, compared to 19.4%, 3.2%, and 0% for patients in the control group (P < .05). Also, at week 24 of LCAP therapy, ACR20, ACR50, and ACR70 were observed in 70%, 50%, and 30% of patients, respectively, which was significantly higher than the 25.8%, 12.9%, and 3.2% of patients in the control group (P < .05). The serum levels of CXCL16 and serotonin were significantly reduced in the LCAP group compared with control group. CONCLUSIONS: This study indicated that LCAP therapy can significantly decrease RA disease activity and is a safe and effective alternative therapy. LCAP therapy significantly reduced serum CXCL16 and serotonin levels, offering a putative mechanism by which it improves the articular symptoms of RA.

Pestalotiopsis-Like Species Causing Gray Blight Disease on Camellia sinensis in China.

Gray blight of tea, caused by several Pestalotiopsis-like species, is one of the most destructive foliar diseases in tea cultivation yet the characteristics of these pathogens have not been confirmed until now. With morphological and multigene phylogenetic analyses, we have identified the gray blight fungi as Pseudopestalotiopsis camelliae-sinensis, Neopestalotiopsis clavispora, and Pestalotiopsis camelliae. Phylogenetic analyses derived from the combined internal transcribed spacer, ß-tubulin, and translation elongation factor 1-α gene regions successfully resolved most of the Pestalotiopsis-like species used in this study with high bootstrap supports and revealed three major clusters representing these three species. Differences in colony appearance and conidia morphology (shape, size, septation, color and length of median cells, and length and number of apical and basal appendages) were consistent with the phylogenetic grouping. Pathogenicity tests validated that all three species isolated from tea leaves were causal agents of gray blight disease on tea plant (Camellia sinensis). This is the first description of the characteristics of the three species Pseudopestalotiopsis camelliae-sinensis, N. clavispora, and Pestalotiopsis camelliae as causal agents of tea gray blight disease in China.

Characterization, Pathogenicity, and Phylogenetic Analyses of Colletotrichum Species Associated with Brown Blight Disease on Camellia sinensis in China.

Brown blight disease caused by Colletotrichum species is a common and serious foliar disease of tea (Camellia sinensis). Fungal isolates from several tea plantations causing typical brown blight symptoms were identified as belonging to the Colletotrichum acutatum species complex and the Colletotrichum gloeosporioides species complex based on morphological characteristics as well as DNA analysis of the internal transcribed spacer (ITS) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH). Colletotrichum acutatum, a new causal agent associated with C. sinensis, showed high phenotypic and genotypic diversity compared with the more commonly reported C. gloeosporioides. Phylogenetic analysis derived from individual and combined ITS and GAPDH sequences clearly clustered C. acutatum and C. gloeosporioides into separate species. Pathogenicity tests validated that both species were causal agents of tea brown blight disease and were highly pathogenic to tea leaves. However, the two groups of C. gloeosporioides with low levels of variability within their ITS and GAPDH regions differed in their virulence. This study reports for the first time the characterization of C. acutatum and C. gloeosporioides causing brown blight disease on tea (Camellia sinensis (L.) O. Kuntze) in China.

Plasmapheresis therapy in combination with TNF-α inhibitor and DMARDs: A multitarget method for the treatment of rheumatoid arthritis.

OBJECTIVE: To evaluate the effects of a multitarget method involving plasmapheresis therapy combined with tumor necrosis factor (TNF)-α inhibitor and disease-modifying antirheumatic drugs (DMARDs) on disease activity parameters in the treatment of active rheumatoid arthritis (RA). METHODS: Sixty-five patients with active RA were divided into two groups according to the treatment administered: the plasmapheresis combination therapy group (Plasmapheresis combination group; 38 cases), in which patients received plasmapheresis therapy along with a TNF-α inhibitor (recombinant human tumor necrosis factor-Fc; rhTNFR:Fc; Etanercept biosimilars) and DMARDs, and a TNF-α inhibitor therapy group (biological agent group; 27 cases), in which patients received a TNF-α inhibitor and DMARDs. Clinical parameters were measured before and at 4 and 24 weeks after treatment. RESULTS: ACR20, ACR50, and ACR70 responses at week 4 were achieved in 84.2%, 78.9%, and 60.5% of the patients in the plasmapheresis combination group, respectively, and 74.1%, 55.6%, and 29.6% of the patients in the biological agent group, respectively. The ACR50 and ACR70 response rates were superior in the former than the latter group (p < 0.05). Similar patterns of statistical significance were observed for ACR20, ACR50, and ACR70 responses at week 24 after the treatment. ACR50 responses were achieved in 84.2% patients and ACR70 responses were achieved in 76.3% patients in the plasmapheresis combination group, and these proportions were better than those in the biological agent group (p < 0.05). CONCLUSIONS: The multitarget method combining plasmapheresis, TNF-α inhibitor, and DMARDs for RA therapy was superior to the combination of TNF-α inhibitor for reducing disease activity parameters in patients with active RA.

Effect of colchicine in prevention of pericardial effusion and atrial fibrillation: a meta-analysis.

Randomized, controlled trials (RCTs) have assessed the effect of colchicine therapy in prevention of pericardial effusion (PE) and atrial fibrillation (AF). However, the effects are still inconclusive. PubMed, Cochrane Library, Google Scholar, and EMBASE database were searched. Primary outcome was the risk of PE and AF. Ten RCTs with 1981 patients and a mean follow-up of 12.6 months were included. Colchicine therapy was not associated with a significantly lower risk of post-operative PE (RR, 0.89; 95 % CI 0.70-1.13; p = 0.33, I (2) = 72.8 %) and AF (RR, 0.77; 95 % CI 0.52-1.13; p = 0.18, I (2) = 47.3 %). However, rates of pericarditis recurrence, symptoms persistence, and pericarditis-related hospitalization were significantly decreased with colchicine treatment. In addition, cardiac tamponade occurrence was similar between groups, and adverse events were significantly higher in the colchicine group. Colchicine may not significantly decrease the post-operative risk of PE and AF. However, only limited studies about patients undergoing cardiac surgery provide data about PE and AF.

A critical domain of Sweet potato chlorotic fleck virus nucleotide-binding protein (NaBp) for RNA silencing suppression, nuclear localization and viral pathogenesis.

RNA silencing is an important mechanism of antiviral defence in plants. To counteract this resistance mechanism, many viruses have evolved RNA silencing suppressors. In this study, we analysed five proteins encoded by Sweet potato chlorotic fleck virus (SPCFV) for their abilities to suppress RNA silencing using a green fluorescent protein (GFP)-based transient expression assay in Nicotiana benthamiana line 16c plants. Our results showed that a putative nucleotide-binding protein (NaBp), but not other proteins encoded by the virus, could efficiently suppress local and systemic RNA silencing induced by either sense or double-stranded RNA (dsRNA) molecules. Deletion mutation analysis of NaBp demonstrated that the basic motif (an arginine-rich region) was critical for its RNA silencing suppression activity. Using confocal laser scanning microscopy imaging of transfected protoplasts expressing NaBp fused to GFP, we showed that NaBp accumulated predominantly in the nucleus. Mutational analysis of NaBp demonstrated that the basic motif represented part of the nuclear localization signal. In addition, we demonstrated that the basic motif in NaBp was a pathogenicity determinant in the Potato virus X (PVX) heterogeneous system. Overall, our results demonstrate that the basic motif of SPCFV NaBp plays a critical role in RNA silencing suppression, nuclear localization and viral pathogenesis.