Intercropping fruit trees in tea plantation improves soil properties and the formation of tea quality components.

Intercropping has been recommended as a beneficial cropping practice for improving soil characteristic and tea quality. However, there is limited research on the effects of intercropping fruit trees on soil chemical properties, soil aggregate structure, and tea quality components. In this study, intercropping fruit trees, specifically loquats and citrus, had a significant impact on the total available nutrients, AMN, and AP in soil. During spring and autumn seasons, the soil large-macroaggregates (>2 mm) proportion increased by 5.93% and 19.03%, as well as 29.23% and 19.14%, respectively, when intercropping loquats and citrus. Similarly, intercropping waxberry resulted in a highest small-macroaggregates (0.25 mm-2 mm) proportion at 54.89% and 77.32%. Soil aggregate stability parameters of the R0.25, MWD, and GMD were generally considered better soil aggregate stability indicators, and significantly improved in intercropping systems. Intercropping waxberry with higher values for those aggregate stability parameters and lower D values, showed a better soil aggregate distribution, while intercropping loquats and citrus at higher levels of AMN and AP in different soil aggregate sizes. As the soil aggregate sizes increased, the AMN and AP contents gradually decreased. Furthermore, the enhanced levels of amino acids were observed under loquat, waxberry, and citrus intercropping in spring, which increased by 27.98%, 27.35%, and 26.21%, respectively. The contents of tea polyphenol and caffeine were lower under loquat and citrus intercropping in spring. These findings indicated that intercropping fruit trees, specifically loquat and citrus, have immense potential in promoting the green and sustainable development of tea plantations.

Leguminous green manure intercropping changes the soil microbial community and increases soil nutrients and key quality components of tea leaves.

Intercropping, a green and sustainable planting pattern, has demonstrated positive effects on plant growth and the soil environment. However, there is currently little research on the influence of intercropping leguminous plants and using them as green manure on the soil environment and tea quality. During the profuse flowering period of Chinese milkvetch, the contents of tea amino acids and soluble sugar in intercropping tea plants with soybean increased by 6.89 and 54.58%. Moreover, there was 27.42% increase in soil ammonium nitrogen and 21.63% increase in available nitrogen. When Chinese milkvetch was returned to soil for 1 month during its profuse flowering period, the soybean and Chinese milkvetch as green manure enhanced tea amino acids and soluble sugar by 9.11 and 33.96%, and soil ammonium nitrogen, nitrate nitrogen and available nitrogen increased by 25.04, 77.84, and 48.90%. Intercropping systems also have positive effects on tea quality components, soil fertility, and soil microbial communities during the profuse flowering period of soybeans and when soybeans with this period were returned to the field for 1 month. Furthermore, the soil fertility index was significantly increased, especially in the intercropping system of tea-soybean-Chinese milkvetch. The soil bacterial community complexity and fungal community interactions were significantly increased. Soil pH, nitrate nitrogen, and available phosphorus were found to be crucial influencing factors on soil microbial communities, specifically bacterial communities. These results highlight the significance of optimizing intercropping systems to improve the soil environment and tea quality components. They also provide a theoretical foundation for promoting the sustainable development of tea plantations.

Causality orientations and spontaneous mental contrasting.

Mental contrasting is a motivational behavior change strategy necessary for strong goal commitment. Meanwhile, general causality orientations are motivational patterns that represent individuals' motivation for behavior change and the reason for their goal commitment. The current study explored whether causality orientations predict spontaneous mental contrasting in Chinese university students. Study 1 investigated whether academic autonomy, control, and amotivated orientations correlate with spontaneous mental contrasting about an important academic goal. The findings of Study 1 reveal that autonomy orientation did not correlate with mental contrasting, whereas control and amotivated orientations were negatively correlated with mental contrasting. Study 2 investigated whether priming autonomy and control orientations, in addition to the neutral condition, would induce spontaneous mental contrasting about an academic goal related to the students' research topic. The results of Study 2 revealed that the autonomy condition orientation did not differ significantly from the controlled orientation condition. However, when compared to the neutral condition, the autonomy condition significantly predicted mental contrasting, whereas the controlled orientation condition did not show any significant difference. In Study 2, the autonomy-oriented participants generated more spontaneous mental contrast than the control orientation and neutral conditions. The findings show that controlled and amotivated orientations predicted negative mental contrasting. As a result, controlled and amotivated students must learn how to use mental contrasting to achieve high levels of goal commitment and achievement. Lastly, the study discussed its implications, limitations, and suggestions for future research.

The effect of intercropping leguminous green manure on theanine accumulation in the tea plant: A metagenomic analysis.

Intercropping is a widely recognised technique that contributes to agricultural sustainability. While intercropping leguminous green manure offers advantages for soil health and tea plants growth, the impact on the accumulation of theanine and soil nitrogen cycle are largely unknown. The levels of theanine, epigallocatechin gallate and soluble sugar in tea leaves increased by 52.87% and 40.98%, 22.80% and 6.17%, 22.22% and 29.04% in intercropping with soybean-Chinese milk vetch rotation and soybean alone, respectively. Additionally, intercropping significantly increased soil amino acidnitrogen content, enhanced extracellular enzyme activities, particularly ß-glucosidase and N-acetyl-glucosaminidase, as well as soil multifunctionality. Metagenomics analysis revealed that intercropping positively influenced the relative abundances of several potentially beneficial microorganisms, including Burkholderia, Mycolicibacterium and Paraburkholderia. Intercropping resulted in lower expression levels of nitrification genes, reducing soil mineral nitrogen loss and N2 O emissions. The expression of nrfA/H significantly increased in intercropping with soybean-Chinese milk vetch rotation. Structural equation model analysis demonstrated that the accumulation of theanine in tea leaves was directly influenced by the number of intercropping leguminous green manure species, soil ammonium nitrogen and amino acid nitrogen. In summary, the intercropping strategy, particularly intercropping with soybean-Chinese milk vetch rotation, could be a novel way for theanine accumulation.

Genome-Wide Association Studies of Biluochun Tea Plant Populations in Dongting Mountain and Comprehensive Identification of Candidate Genes Associated with Core Agronomic Traits by Four Analysis Models.

The elite germplasm resources are key to the beautiful appearance and pleasant flavor of Biluochun tea. We collected and measured the agronomic traits of 95 tea plants to reveal the trait diversity and breeding value of Biluochun tea plant populations. The results revealed that the agronomic traits of Biluochun tea plant populations were diverse and had high breeding value. Additionally, we resequenced these tea plant populations to reveal genetic diversity, population structure, and selection pressure. The Biluochun tea plant populations contained two groups and were least affected by natural selection based on the results of population structure and selection pressure. More importantly, four non-synonymous single nucleotide polymorphisms (nsSNPs) and candidate genes associated with (-)-gallocatechin gallate (GCG), (-)-gallocatechin (GC), and caffeine (CAF) were detected using at least two GWAS models. The results will promote the development and application of molecular markers and the utilization of elite germplasm from Biluochun populations.

Minigene Assay as an Effective Molecular Diagnostic Strategy in Determining the Pathogenicity of Noncanonical Splice-Site Variants in FLCN.

Primary spontaneous pneumothorax (PSP) or pulmonary cyst is one of the manifestations of Birt-Hogg-Dubé syndrome, which is caused by pathogenic variants in FLCN gene. Genetic testing in patients with PSP identifies a certain number of missense or intronic variants. These variants are usually considered as variants of uncertain significance, whose functional interpretations pose a challenge in clinical genetics. To improve recognition of pathogenic splice-altering variants in FLCN gene, computational tools are used to prioritize potential splice-altering variants and then a hybrid minigene assay is performed to verify the RNA splicing pattern. Herein, variants in FLCN exon 11 and its flanking sequence are focused. Eight variants detected in 11 patients with PSP are evaluated, and six variants are prioritized by in silico tools as potential splice-altering variants of uncertain significance. Four variants (c.1177-5_1177-3delCTC, c.1292_1300+4del, c.1300+4C>T, and c.1300+5G>A) are demonstrated by minigene assay to alter RNA splicing of FLCN, and the last three of them are novel. RT-PCR of patient-derived RNA gives consistent results. Genotype-phenotype correlation analysis in patients with PSP with these variants demonstrates good concordance. Our results underline the importance of RNA analysis, which could provide molecular evidence for pathogenicity of a variant, and provide essential information for the clinical interpretation of variants. Combining the clinical information, a definitive diagnosis could be made.

Development of Novel Lipid-Based Formulations for Water-Soluble Vitamin C versus Fat-Soluble Vitamin D3.

The aim of this study was to develop a facile and novel lipid-based formulation of vitamin C and vitamin D3. Liposomes loaded with vitamin C and D3 were characterized using transmission electron microscopy (TEM) and zeta potential measurements for evaluating morphology, particle size and physical stability. HPLC was employed to quantify the content of vitamin C and vitamin D3 in their liposomal forms. The UHPLC analysis of the lipid-based vitamin formulation is an easy and rapid method for the characterization as well as the quantification of all components. In addition, encapsulation efficiency, vitamin loading and stability analysis were performed by the UHPLC method, in order to evaluate the reliability of the optimized lipid-based formulation. The TEM results provided key support for the core type of liposome structure in the formulations, whereas the HPLC results indicated that the liposomal vitamin C and D3 systems were homogeneous, and did not undergo phase separation. Taken together, the results demonstrate that liposomal encapsulated vitamins (vitamin C and D3) possess a unilamellar vesicle morphology with uniform particle size, despite differences in the hydrophile-lipophile profiles of the vitamins. The highly efficient encapsulation properties of such liposomal constructs are proposed to contribute to enhanced vitamin bioavailability.

CsMYB Transcription Factors Participate in Jasmonic Acid Signal Transduction in Response to Cold Stress in Tea Plant (Camellia sinensis).

Low-temperature stress is an increasing problem for the cultivation of tea (Camellia sinensis), with adverse effects on plant growth and development and subsequent negative impacts on the tea industry. Methyl jasmonate (MeJA), as a plant inducer, can improve the cold-stress tolerance in tea plants. R2R3-MYB transcription factors (TFs) are considered potentially important regulators in the resistance to cold stress in plants. However, the molecular mechanisms, by which MYB TFs via the jasmonic acid pathway respond to cold stress in the tea plant, remain unknown. In this study, physiological and biochemical assays showed that exogenous MeJA application could effectively promote ROS scavenging in the tea plant under cold stress, maintaining the stability of the cell membrane. Sixteen R2R3-MYB TFs genes were identified from the tea plant genome database. Quantitative RT-PCR analysis showed that three CsMYB genes were strongly induced under a combination of MeJA and cold-stress treatment. Subcellular localization assays suggest CsMYB45, CsMYB46, and CsMYB105 localized in the nucleus. Exogenous MeJA treatment enhanced the overexpression of CsMYB45, CsMYB46, and CsMYB105 in E. coli and improved the growth and survival rates of recombinant cells compared to an empty vector under cold stress. Yeast two-hybrid and bimolecular fluorescence complementation experiments confirmed that CsMYB46 and CsMYB105 interacted with CsJAZ3, CsJAZ10, and CsJAZ11 in the nucleus. Taken together, these results highlight that CsMYB45, CsMYB46, and CsMYB105 are not only key components in the cold-stress signal response pathway but also may serve as points of confluence for cold stress and JA signaling pathways. Furthermore, our findings provide new insight into how MYB TFs influence cold tolerance via the jasmonic acid pathway in tea and provide candidate genes for future functional studies and breeding.

Genome-Wide Analysis of the TCP Gene Family and Their Expression Pattern Analysis in Tea Plant (Camellia sinensis).

TEOSINTE BRANCHED1/CYCLOIDEA/PCF (TCP) transcription factors TEOSINTE BRANCHED1/CYCLOIDEA/PCF have been suggested to control the cell growth and proliferation in meristems and lateral organs. A total of 37 CsTCP genes were identified and divided into two classes, class I (PCF, group 1) and class II (CIN CYC/TB1, groups 2, and 3). The residues of TEOSINTE BRANCHED1/CYCLOIDEA/PCF of Camellia sinensis (Tea plant) (CsTCP) proteins between class I and class II were definitely different in the loop, helix I, and helix II regions; however, eighteen conserved tandem was found in bHLH. There are a large number of CsTCP homologous gene pairs in three groups. Additionally, most CsTCP proteins have obvious differences in motif composition. The results illuminated that CsTCP proteins in different groups are supposed to have complementary functions, whereas those in the same class seem to display function redundancies. There is no relationship between the number of CsTCP gene members and genome size, and the CsTCP gene family has only expanded since the divergence of monocots and eudicots. WGD/segmental duplication played a vital role in the expansion of the CsTCP gene family in tea plant, and the CsTCP gene family has expanded a lot. Most CsTCP genes of group 1 are more widely and non-specifically expressed, and the CsTCP genes of group 2 are mainly expressed in buds, flowers, and leaves. Most genes of group 1 and some genes of group 2 were up-/downregulated in varying degrees under different stress, CsTCP genes of group 3 basically do not respond to stress. TCP genes involved in abiotic stress response mostly belong to PCF group. Some CsTCP genes may have the same function as the homologous genes in Arabidopsis, but there is functional differentiation.

CsAAP7.2 is involved in the uptake of amino acids from soil and the long-distance transport of theanine in tea plants (Camellia sinensis L.).

Tea plant roots can uptake both inorganic nitrogen (NH4+ and NO3-) and organic nitrogen (amino acids) from the soil. These amino acids are subsequently assimilated into theanine and transported to young shoots through the xylem. Our previous study showed that CsLHT1 and CsLHT6 transporters take up amino acids from the soil, and CsAAPs participate in the transport of theanine. However, whether other amino acid transporters are involved in this process remains unknown. In this study, we identified two new CsAAPs homologous to CsAAP7, named CsAAP7.1 and CsAAP7.2. Heterologous expression of CsAAP7.1 and CsAAP7.2 in the yeast mutant 22Δ10α showed that CsAAP7.2 had the capacity to transport theanine and other amino acids, whereas CsAAP7.1 had no transport activity. Transient expression of the CsAAP7.2-GFP fusion protein in tobacco leaf epidermal cells confirmed its localization to the endoplasmic reticulum. Tissue-specific analysis showed that CsAAP7.2 was highly expressed in roots and stems. In addition, CsAAP7.2 overexpression lines were more sensitive to high concentrations of theanine due to the high accumulation of theanine in seedlings. Taken together, these findings suggested that CsAAP7.2 plays an important role in the uptake of amino acids from soil and the long-distance transport of theanine. These results provide valuable tools for nitrogen nutrition studies and enrich our understanding of theanine transport in tea plants.

CsCuAOs and CsAMADH1 Are Required for Putrescine-Derived γ-Aminobutyric Acid Accumulation in Tea.

Polyamines are a potential source of γ-aminobutyric acid (GABA) in plants under abiotic stress. However, studies on GABA enrichment in tea mostly focus on the GABA shunt, while the correlation between polyamine degradation and GABA formation in tea is largely unknown. In this study, tea plants responded to exogenous putrescine, resulting in a significant increase in GABA content, while the glutamate level did not change. At the same time, five copper-containing amine oxidase (CuAO) and eight aminoaldehyde dehydrogenase (AMADH) genes involved in the putrescine-derived GABA pathway were identified from the Tea Plant Information Archive. Expression analysis indicated that CsCuAO1, CsCuAO3 as well as CsAMADH1 were induced to play an important function in response to exogenous putrescine. Thus, the three genes were cloned and the catalytic efficiency of soluble recombinant proteins was determined. CsCuAOs and CsAMADH1 exhibited indispensable functions in the GABA production from putrescine in vitro. Subcellular localization assays indicated that CsAMADH1 was localized in plastid, while both CsCuAO1 and CsCuAO3 were localized in peroxisome. In addition, the synergistic effects of CsCuAOs and CsAMADH1 were investigated by a transient co-expression system in Nicotiana benthamiana. Our data suggest that these three genes regulate the accumulation of GABA in tea by participating in the polyamine degradation pathway and improve the content of GABA in tea to a certain extent. The results will greatly contribute to the production of GABA tea.

Functional analysis of variants in DMD exon/intron 10 predicted to affect splicing.

Duchenne muscular dystrophy (DMD, MIM #310200) and Becker muscular dystrophy (BMD, MIM #300376) are X-linked recessive hereditary diseases caused by pathogenic variants in the DMD gene. Genetic testing of DMD identifies a certain number of variants of uncertain clinical significance (VUS) whose functional interpretations pose a challenge for gene-based diagnosis. To improve the accuracy of variant interpretation in public mutation repositories, we used computational tools to prioritize VUS and developed a cell-based minigene assay to confirm aberrant splicing. Using this procedure, we evaluated rare variants in exon and intron 10 of the DMD gene. We demonstrated that 16 variants, including both canonical and non-canonical splice sites, altered RNA splicing in variable patterns. Using the example of exon and intron 10 of the DMD gene, we demonstrated the utility of the in vitro minigene assay in the effective assessment of the spliceogenic effect for VUS identified in clinical practice and underlined the necessity of precise variant classification. This is the first systematic characterization of DMD splicing variants, besides, through our study, some undetermined variants are demonstrated to be pathogenic by altering RNA splicing of DMD.

Distribution of trace metals in a soil-tea leaves-tea infusion system: characteristics, translocation and health risk assessment.

The effects of metal pollution on tea are of great concern to consumers. We apply Geographic information systems technology to study the distribution of heavy metal elements in tea plantation ecosystems in Jiangsu Province, explore the relationships among metals in the soil, tea leaves and tea infusions, and assess the human safety risks of metals. The concentrations of nine metals in a soil-tea leaves-tea infusion system were studied at 100 randomly selected tea plantations in Jiangsu Province, China. Concentrations of selected metals, zinc (Zn), nickel (Ni), manganese (Mn), chromium (Cr) and copper (Cu), were quantified using an inductively coupled plasma-optical emission spectrometer (ICP-OES), and cadmium (Cd), arsenic (As), plumbum (Pb) and mercury (Hg) were quantified using inductively coupled plasma-mass spectrometry (ICP-MS). Arc-Map 10.3 was used for the spatial analysis of metals in soil, tea leaves and tea infusions. We found that the contents of Mn, Ni and Zn are high level in soil, tea leaves and tea infusions. The Mn level showed a spatial distribution pattern with greater concentrations at the junction of Nanjing and Yangzhou, southwest of Changzhou and west of Suzhou. The hazard index (HI) values in north-central Nanjing, southern Suzhou, southwestern Changzhou and northern Lianyungang were relatively greater. The Zn, Ni, Mn, Cr and Cu levels in the soil-tea infusion system were 17.3, 45.5, 54.5, 1.5 and 14.3%, respectively. The order of the leaching rates of the elements was Ni > Cr > Zn > Mn > Cu. The relative contribution ratios of HI were in the order of Mn > Ni > Cu > Zn > Cr > Pb > Cd > As > Hg. In tea infusions, the Mn level has the greatest potential health risks to consumers. Moreover, using Csoil it was inferred that the safety thresholds of Zn, Ni, Mn, Cr and Cu in soil were 27,700, 50, 1230, 493,000 and 16,800 mg L-1, respectively. The content of heavy metals in soil and tea varies greatly in different regions of Jiangsu Province, 92% of the soil has heavy metal content that meets the requirements of pollution-free tea gardens, 91% of tea samples met the requirements of green food tea. The thresholds for Ni (50 mg L-1) and Mn (1230 mg L-1) can be used as maximum limits in tea plantation soils. The consumption of tea infusions did not pose metal-related risks to human health.

The effects of tea plants-soybean intercropping on the secondary metabolites of tea plants by metabolomics analysis.

BACKGROUND: Intercropping, especially with legumes, as a productive and sustainable system, can promote plants growth and improves the soil quality than the sole crop, is an essential cultivation pattern in modern agricultural systems. However, the metabolic changes of secondary metabolites and the growth in tea plants during the processing of intercropping with soybean have not been fully analyzed. RESULTS: The secondary metabolomic of the tea plants were significant influence with intercropping soybean during the different growth stages. Especially in the profuse flowering stage of intercropping soybean, the biosynthesis of amino acids was significantly impacted, and the flavonoid biosynthesis, the flavone and flavonol biosynthesis also were changed. And the expression of metabolites associated with amino acids metabolism, particularly glutamate, glutamine, lysine and arginine were up-regulated, while the expression of the sucrose and D-Glucose-6P were down-regulated. Furthermore, the chlorophyll photosynthetic parameters and the photosynthetic activity of tea plants were higher in the tea plants-soybean intercropping system. CONCLUSIONS: These results strengthen our understanding of the metabolic mechanisms in tea plant's secondary metabolites under the tea plants-soybean intercropping system and demonstrate that the intercropping system of leguminous crops is greatly potential to improve tea quality. These may provide the basis for reducing the application of nitrogen fertilizer and improve the ecosystem in tea plantations.

An ancient whole-genome duplication event and its contribution to flavor compounds in the tea plant (Camellia sinensis).

Tea, coffee, and cocoa are the three most popular nonalcoholic beverages in the world and have extremely high economic and cultural value. The genomes of four tea plant varieties have recently been sequenced, but there is some debate regarding the characterization of a whole-genome duplication (WGD) event in tea plants. Whether the WGD in the tea plant is shared with other plants in order Ericales and how it contributed to tea plant evolution remained unanswered. Here we re-analyzed the tea plant genome and provided evidence that tea experienced only WGD event after the core-eudicot whole-genome triplication (WGT) event. This WGD was shared by the Polemonioids-Primuloids-Core Ericales (PPC) sections, encompassing at least 17 families in the order Ericales. In addition, our study identified eight pairs of duplicated genes in the catechins biosynthesis pathway, four pairs of duplicated genes in the theanine biosynthesis pathway, and one pair of genes in the caffeine biosynthesis pathway, which were expanded and retained following this WGD. Nearly all these gene pairs were expressed in tea plants, implying the contribution of the WGD. This study shows that in addition to the role of the recent tandem gene duplication in the accumulation of tea flavor-related genes, the WGD may have been another main factor driving the evolution of tea flavor.

Meta-analysis of the effect of expression of MYB transcription factor genes on abiotic stress.

BACKGROUND: MYB proteins are a large group of transcription factors. The overexpression of MYB genes has been reported to improve abiotic stress tolerance in plant. However, due to the variety of plant species studied and the types of gene donors/recipients, along with different experimental conditions, it is difficult to interpret the roles of MYB in abiotic stress tolerance from published data. METHODS: Using meta-analysis approach, we investigated the plant characteristics involved in cold, drought, and salt stress in MYB-overexpressing plants and analyzed the degrees of influence on plant performance by experimental variables. RESULTS: The results show that two of the four measured plant parameters in cold-stressed plants, two of the six in drought-stressed, and four of the 13 in salt-stressed were significantly impacted by MYB overexpression by 22% or more, and the treatment medium, donor/recipient species, and donor type significantly influence the effects of MYB-overexpression on drought stress tolerance. Also, the donor/recipient species, donor type, and stress duration all significantly affected the extent of MYB-mediated salt stress tolerance. In summary, this study compiles and analyzes the data across studies to help us understand the complex interactions that dictate the efficacy of heterologous MYB expression designed for improved abiotic stress tolerance in plants.

Genome-Wide Identification and Expression Analysis of the NRAMP Family Genes in Tea Plant (Camellia sinensis).

The natural resistant-associated macrophage protein (NRAMP) is a kind of integral membrane transporter which could function on a wide range of divalent metal ions in plants. Little is known about the NRAMP family in Camellia sinensis. In this study, 11 NRAMP genes were identified from the tea plant genome. Phylogenetic analysis showed that the 11 CsNRAMP proteins were split into two groups. The proteins of group 1 contained the conserved motif 6 (GQSSTxTG), while most proteins in group 2 (excepting CsNRAMP7 and CsNRAMP10) contained the conserved residues of motif 6 and motif 2 (GQFIMxGFLxLxxKKW). The number of amino acids in coding regions of 11 CsNRAMP genes ranged from 279-1373, and they contained 3-12 transmembrane domains. Quantitative RT-PCR analysis showed that G1 genes, CsNRAMP3, CsNRAMP4, and CsNRAMP5, were extraordinarily expressed in roots, while G2 genes showed higher expression levels in the stems and leaves. The expression levels of CsNRAMPs in roots and leaves were detected to assess their responses to Pb treatment. The results indicated that CsNRAMPs were differentially regulated, and they might play a role in Pb transportation of tea plant. Subcellular localization assay demonstrated that CsNRAMP2 and CsNRAMP5 fused proteins were localized in the plasma membrane. Overall, this systematic analysis of the CsNRAMP family could provide primary information for further studies on the functional roles of CsNRAMPs in divalent metal transportation in tea plants.

A traditional medicine, respiratory detox shot (RDS), inhibits the infection of SARS-CoV, SARS-CoV-2, and the influenza A virus in vitro.

BACKGROUND: The ongoing global pandemic of coronavirus disease 2019 (COVID-19) has resulted in the infection of over 128 million people and has caused over 2.8 million deaths as of April 2021 in more than 220 countries and territories. Currently, there is no effective treatment for COVID-19 to reduce mortality. We investigated the potential anti-coronavirus activities from an oral liquid of traditional medicine, Respiratory Detox Shot (RDS), which contains mostly herbal ingredients traditionally used to manage lung diseases. RESULTS: Here we report that RDS inhibited the infection of target cells by lenti-SARS-CoV, lenti-SARS-CoV-2, and hybrid alphavirus-SARS-CoV-2 (Ha-CoV-2) pseudoviruses, and by infectious SARS-CoV-2 and derived Ha-CoV-2 variants including B.1.1.7, B.1.351, P.1, B.1.429, B.1.2, B.1.494, B.1.1.207, B.1.258, and B.1.1.298. We further demonstrated that RDS directly inactivates the infectivity of SARS-CoV-2 virus particles. In addition, we found that RDS can also block the infection of target cells by Influenza A virus. CONCLUSIONS: These results suggest that RDS may broadly inhibit the infection of respiratory viruses.

Preparation and characterization of cyclic citrullinated peptide-immobilized latex beads for measurement of anti-citrillinated protein antibody through latex particle-enhanced turbidimetric immunoassay.

The anti-citrillinated protein antibody (ACPA) plays an important role in early diagnosis of rheumatoid arthritis (RA), and is usually detected by using cyclic citrullinated peptide (CCP) as antigen. The ACPA against CCP test is usually performed utilizing enzyme-linked immunosorbent assay (ELISA), but the ELISA is expensive and time-consuming. Here, latex particle-enhanced turbidimetric immunoassay (LTIA) based on CCP-immobilized latex bead was proposed for fast measurements of ACPA of RA patients. CCP-immobilized latex bead was fabricated through three methods, including direct coupling, overall coupling and layer by layer coupling. According to the optimized experiments, layer-by-layer coupling was the best method with advantages of time-saving, simple operation and good repeatability. In addition, a spacer arm of appropriate length between latex beads and CCP could avoid stereoscopic obstacles and make ACPA closer to CCP. The CCP-immobilized latex bead based on layer by layer coupling (CCP-LB-LLC) was used for assembling the homemade kit, which was applied in fast measurements of ACPA through LTIA. The homemade kit possessed a low limit of detection (0.2 U/mL) and an acceptable the batch-to-batch reproducibility. In addition, the homemade kit can be stored at 4 °C for at least one month. When used to detect 20 clinical samples, the results of homemade kit were consistent with commercial ELISA. Furthermore, LTIA based on the homemade kit was simpler and cheaper than ELISA. These results demonstrated that the homemade kit could be useful for diagnosis of RA patients.

Effects of GABA on the polyphenol accumulation and antioxidant activities in tea plants (Camellia sinensis L.) under heat-stress conditions.

Polyphenols are important active components in tea plants, which have strong biological activity and antioxidant activity. A certain degree of stress or exogenous substances can significantly increase the content of polyphenols in plants. γ-Aminobutyric acid (GABA), a natural functional amino acid, was used to study whether exogenous GABA can increase the content of polyphenols and enhance antioxidant activity in tea plants under heat-stress conditions. The results showed that the content of GABA was positively correlated with the content of polyphenols (r = 0.649), especially with the content of total catechins (r = 0.837). Most of the related genes encoding flavonoid metabolism (PAL, C4H, 4CL, CHS, CHI, F3H, F3'H, F3'5'H, DFR, LAR, ANS, ANR and FLS) as well as enzyme activities (PAL, C4H and 4CL) were upregulated. In addition, the activities of antioxidant enzymes were induced under heat-stress conditions. However, 3-mercaptopropionic acid (3-MPA), an inhibitor of GABA synthesis, exhibited opposite results under heat-stress conditions compared with GABA treatment. These results indicated that GABA plays a key role in the accumulation of polyphenols and the upregulation of the antioxidant system in tea plants under heat-stress conditions.