The diversity of endophytic fungi in Tartary buckwheat (Fagopyrum tataricum) and its correlation with flavonoids and phenotypic traits.

Tartary buckwheat (Fagopyrum tataricum) is a significant medicinal crop, with flavonoids serving as a crucial measure of its quality. Presently, the artificial cultivation of Tartary buckwheat yields low results, and the quality varies across different origins. Therefore, it is imperative to identify an effective method to enhance the yield and quality of buckwheat. Endophytic fungi reside within plants and form a mutually beneficial symbiotic relationship, aiding plants in nutrient absorption, promoting host growth, and improving secondary metabolites akin to the host. In this study, high-throughput sequencing technology was employed to assess the diversity of endophytic fungi in Tartary buckwheat. Subsequently, a correlation analysis was performed between fungi and metabolites, revealing potential increases in flavonoid content due to endophytic fungi such as Bipolaris, Hymenula, and Colletotrichum. Additionally, a correlation analysis between fungi and phenotypic traits unveiled the potential influence of endophytic fungi such as Bipolaris, Buckleyzyma, and Trichosporon on the phenotypic traits of Tartary buckwheat. Notably, the endophytic fungi of the Bipolaris genus exhibited the potential to elevate the content of Tartary buckwheat metabolites and enhance crop growth. Consequently, this study successfully identified the resources of endophytic fungi in Tartary buckwheat, explored potential functional endophytic fungi, and laid a scientific foundation for future implementation of biological fertilizers in improving the quality and growth of Tartary buckwheat.

Serendipita indica Promotes the Growth of Tartary Buckwheat by Stimulating Hormone Synthesis, Metabolite Production, and Increasing Systemic Resistance.

The main objective of this study was to investigate the influence of Serendipita indica on the growth of Tartary buckwheat plants. This study highlighted that the roots of Tartary buckwheat can be colonized by S. indica and that this fungal endophyte improved plants height, fresh weight, dry weight, and grain yield. In the meantime, the colonization of S. indica in Tartary buckwheat leaves resulted in elevated levels of photosynthesis, plant hormone content, antioxidant enzyme activity, proline content, chlorophyll content, soluble sugars, and protein content. Additionally, the introduction of S. indica to Tartary buckwheat roots led to a substantial rise in the levels of flavonoids and phenols found in the leaves and seeds of Tartary buckwheat. In addition, S. indica colonization reduced the content of malondialdehyde and hydrogen peroxide when compared to non-colonized plants. Importantly, the drought tolerance of Tartary buckwheat plants is increased, which benefits from physiology and bio-chemical changes in plants after S. indica colonized. In conclusion, we have shown that S. indica can improve systematic resistance and promote the growth of Tartary buckwheat by enhancing the photosynthetic capacity of Tartary buckwheat, inducing the production of IAA, increasing the content of secondary metabolites such as total phenols and total flavonoids, and improving the antioxidant enzyme activity of the plant.

The Regulatory Functions of the Multiple Alternative Sigma Factors RpoE, RpoHI, and RpoHII Depend on the Growth Phase in Rhodobacter sphaeroides.

Bacterial growth, under laboratory conditions or in a natural environment, goes through different growth phases. Some gene expressions are regulated with respect to the growth phase, which allows bacteria to adapt to changing conditions. Among them, many gene transcriptions are controlled by RpoHI or RpoHII in Rhodobacter sphaeroides. In a previous study, it was proven that the alternative sigma factors, RpoE, RpoHI, and RpoHII, are the major regulators of oxidative stress. Moreover, the growth of bacteria reached a stationary phase, and following the outgrowth, rpoE, rpoHI, and rpoHII mRNAs increased with respect to the growth phase. In this study, we demonstrated the regulatory function of alternative sigma factors in the rsp_0557 gene. The gene rsp_0557 is expressed with respect to the growth phase and belongs to the RpoHI/RpoHII regulons. Reporter assays showed that the antisigma factor ChrR turns on or over the RpoE activity to regulate rsp_0557 expression across the growth phase. In the exponential phase, RpoHII and sRNA Pos19 regulate the expression of rsp_0557 to an appropriate level under RpoE control. In the stationary phase, RpoHI and Pos19 stabilize the transcription of rsp_0557 at a high level. During outgrowth, RpoHI negatively regulates the transcription of rsp_0557. Taken together, our data indicate that these regulators are recruited by cells to adapt to or survive under different conditions throughout the growth phase. However, they still did not display all of the regulators involved in growth phase-dependent regulation. More research is still needed to learn more about the interaction between the regulators and the process of adapting to changed growth conditions and environments.

Diversity, chemical constituents and biological activities of endophytic fungi from Alisma orientale (Sam.) Juzep.

The dried tuber of Alisma orientale (Sam.) Juzep. (AOJ) is a traditional Chinese medicine with high medicinal value. The endophytic fungi of medicinal plants are a treasure house of natural compounds. However, there is a lack of research on the diversity and biological activity of endophytic fungi of AOJ. In this study, high-throughput sequencing technology was used to study the diversity of endophytic fungi in the roots and stems of AOJ, and endophytic fungi with a high output of phenols and flavonoids were screened by chromogenic reaction, and the antioxidant and antibacterial activities and chemical constituents of crude extracts of their fermentation broth were studied. A total of 3,426 amplicon sequence variants (ASVs) belonging to 9 phyla, 27 classes, 64 orders, 152 families, and 277 genera were identified from AOJ. There were significant differences in the endophytic fungal communities of AOJ roots and stems, as well as in the endophytic fungal communities of triangular AOJ and circular AOJ. In addition, 31 strains of endophytic fungi were isolated from AOJ, of which 6 strains had good antioxidant and antibacterial activities. The crude extract of YG-2 had the strongest free radical scavenging ability and bacteriostatic ability, and its IC50 DPPH, IC50 ABTS, and IC50⋅OH values were 0.009 ± 0.000 mg/mL, 0.023 ± 0.002 mg/mL, and 0.081 ± 0.006 mg/mL, respectively. The results of LC-MS showed that the main component of the crude extract of YG-2 was caffeic acid (10.12 µmol/g). Overall, the results of this study preliminarily elucidated the diversity and community composition of endophytic fungi of AOJ, indicating that AOJ endophytic fungi have abundant secondary metabolites and good antioxidant and antibacterial activities. This study provides an important reference for further research, development and utilization of AOJ endophytic fungi and a theoretical basis for the further development of the endophytic fungus YG-2 (Chaetomium globosum) as a source of antioxidants.

The Tartary buckwheat bHLH gene ALCATRAZ contributes to silique dehiscence in Arabidopsis thaliana.

Tartary buckwheat is popular because of its rich nutrients. However, the difficulty in shelling restricts food production. The gene ALCATRAZ (AtALC) plays a key role in silique dehiscence in Arabidopsis thaliana. In this study, an atalc mutant was obtained by CRISPR/Cas9, and a FtALC gene homologous to AtALC was complemented into the atalc mutant to verify its function. Phenotypic observations showed that three atalc mutant lines did not dehiscence, while ComFtALC lines recovered the dehiscence phenotype. The contents of lignin, cellulose, hemicellulose, and pectin in the siliques of all the atalc mutant lines were significantly higher than those in the wild-type and ComFtALC lines. Moreover, FtALC was found to regulate the expression of cell wall pathway genes. Finally, the interaction of FtALC with FtSHP and FtIND was verified by yeast two-hybrid, bimolecular fluorescent complimentary (BIFC) and firefly luciferase completion imaging assays (LCIs). Our findings enrich the silique regulatory network and lay the foundation for the cultivation of easily shelled tartary buckwheat varieties.

Preparation, structural analysis and physicochemical properties of the Cordyceps cicadae exopolysaccharide iron complex.

In this study, EPS-Fe(III) complexes were synthesized, and their structural characteristics, thermal stability, antioxidant activity and digestive properties were evaluated. The content of iron in the EPS-Fe(III) complex was 6.34 ± 1.43 %. The absorbance bands of EPS and EPS-Fe(III) complexes were easily changed, indicating that iron ions can interact with the hydroxyl or carboxyl groups of EPS. Energy spectrometric analysis showed that a strong iron signal was observed in the EPS-Fe(III) complex. The IC50 values of the EPS-Fe(III) complex for DPPH, hydroxyl radical and ABTS were 1.52 mg/mL, 2.63 mg/mL and 1.20 mg/mL, respectively. Under oxidative stress, EPS-Fe(III) can prolong the lifespan of nematodes through the DAF-16 and SKN-1 pathways. Under the condition of gastric juice and intestinal juice, the iron content released from artificial intestinal juice reached 66 %. In addition, the negative effect of trypsin or polyphenols on the solubility of iron in EPS-Fe(III) digestive solution was lower than that in ferric chloride digestive solution. In conclusion, the EPS-Fe(III) complex can be used as a new type of iron supplement, which has good antioxidant activity, high stability and good water solubility.

ABA and SA Participate in the Regulation of Terpenoid Metabolic Flux Induced by Low-Temperature within Conyza blinii.

Under dry-hot valley climates, Conyza blinii (also known as Jin Long Dan Cao), suffers from nocturnal low-temperature stress (LTS) during winter. Here, to investigate the biological significance of terpenoid metabolism during LTS adaptation, the growth state and terpenoid content of C. blinii under different LTS were detected, and analyzed with the changes in phytohormone. When subjected to LTS, the results demonstrated that the growth activity of C. blinii was severely suppressed, while the metabolism activity was smoothly stimulated. Meanwhile, the fluctuation in phytohormone content exhibited three different physiological stages, which are considered the stress response, signal amplification, and stress adaptation. Furthermore, drastic changes occurred in the distribution and accumulation of terpenoids, such as blinin (diterpenoids from MEP) accumulating specifically in leaves and oleanolic acid (triterpenoids from MVA) accumulating evenly and globally. The gene expression of MEP and MVA signal transduction pathways also changes under LTS. In addition, a pharmacological study showed that it may be the ABA-SA crosstalk driven by the LTS signal, that balances the metabolic flux in the MVA and MEP pathways in an individual manner. In summary, this study reveals the different standpoints of ABA and SA, and provides a research foundation for the optimization of the regulation of terpenoid metabolic flux within C. blinii.

Ligusticum chuanxiong Hort as a medicinal and edible plant foods: Antioxidant, anti-aging and neuroprotective properties in Caenorhabditis elegans.

Ligusticum chuanxiong Hort. (CX) is a medicinal and edible plant including a variety of active substances, which may be an available resource for the treatment of related diseases. To expand the medicinal uses of CX, this study aims to explore the antioxidant, anti-aging and neuroprotective effects of the Ligusticum chuanxiong leaves (CXL) and rhizome (CXR) extracts. We first characterize CX phytochemical spectrum by LC-MS as well as antioxidant capacity. Acute toxicity, anti-oxidative stress capacity, lifespan and healthspan was evaluated in C elegans N2. Neuroprotective effect was evaluated in vitro and in vivo (C elegans CL4176 and CL2355). In this study, we detected 74 and 78 compounds from CXR and CXL, respectively, including phthalides, alkaloids, organic acids, terpenes, polyphenols and others. Furthermore, we found that CXs not only protect against oxidative stress, but also prolong the lifespan, alleviate lipofuscin, malondialdehyde (MDA) and reactive oxygen species (ROS) accumulation, and improve movement level, antioxidant enzyme activity in C elegans N2. However, only CXR reduced the ß-amyloid peptide (Aß)-induced paralysis phenotype in CL4176s and alleviated chemosensory behavior dysfunction in CL2355s. In addition, CXR treatment reduced the production of Aß and ROS, enhanced SOD activity in CL4176s. The possible mechanism of anti-aging of CXL and CXR is to promote the expression of related antioxidant pathway genes, increase the activity of antioxidant enzymes, and reduce the accumulation of ROS, which is dependent on DAF-16 and HSF-1 (only in CXR). CXR was able to activate antioxidase-related (sod-3 and sod-5) and heat shock protein genes (hsp-16.1 and hsp-70) expression, consequently ameliorating proteotoxicity related to Aß aggregation. In summary, these findings demonstrate the antioxidant, anti-aging and neuroprotective (only in CXR) activities of the CX, which provide an important pharmacological basis for developing functional foods and drugs to relieve the symptoms of aging and AD. However, the material basis of neuroprotective activity and antiaging effects need to be elucidated, and the relationship between these activities should also be clarified in future studies.

The endophytic fungus Penicillium oxalicum isolated from Ligusticum chuanxiong Hort possesses DNA damage-protecting potential and increases stress resistance properties in Caenorhabditis elegans.

The chemical composition and antioxidant activity of extracts (POE) of Penicillium oxalate isolated from Ligusticum chuanxiong Hort have been investigated. However, the biological activity of POE is limited, and its antioxidant, stress resistance and DNA protection effects in vivo are unclear. The current study aims to explore the beneficial effects of POE on DNA damage protection in pBR322 plasmid and lymphocytes and stress resistance in Caenorhabditis elegans. The results showed that POE increased the survival rate of C. elegans under 35°C, UV and H2O2 stress, attenuated ROS and MDA accumulation, and enhanced the activity of some important enzymes (SOD, CTA, and GSH-PX). In addition, the POE-mediated stress resistance involved the upregulation of the expression of the sod-3, sod-5, gst-4, ctl-1, ctl-2, daf-16, hsp-16.1, hsp-16.2, and hsf-1 genes and acted dependently on daf-16 and hsf-1 rather than skn-1. Moreover, POE also reduced lipofuscin levels, but did not prolong the lifespan or damage the growth, reproduction and locomotion of C. elegans. Furthermore, POE showed a protective effect against DNA scission in the pBR322 plasmid and lymphocytes. These results suggested that P. oxalate extracts have significant anti-stress and DNA protection potential and could be potential drug candidates in the pharmaceutical field, thus greatly broadening the understanding of the biological effects of the endophytic fungus P. oxalate.

CqZF-HD14 enhances drought tolerance in quinoa seedlings through interaction with CqHIPP34 and CqNAC79.

Drought stress is a key agricultural problem that restricts plant development and crop yield. Research on quinoa (Chenopodium quinoa), a nutrient-rich crop with strong stress resistance, has been limited in terms of the molecular regulation of its adaptation to drought stress. This study identified the zinc finger-homeodomain (ZF-HD) family in quinoa and a drought-responsive Chenopodium quinoa ZF-HD14 (CqZF-HD14) through expression profiles. Transient overexpression of CqZF-HD14 promotes photosynthetic pigment accumulation under drought stress, strengthens the antioxidant system, and in turn enhances drought tolerance. Comprehensive genome-wide family analysis and expression profiling identified CqNAC79 and CqHIPP34 regulated by CqZF-HD14, and their interactions were further determined by bimolecular fluorescence complementation (BIFC). Moreover, physiological and biochemical analyses and transient overexpression also revealed that CqNAC79 and CqHIPP34 resist drought by promoting the accumulation of photosynthetic pigments and maintaining antioxidant capacity under drought stress. The synergistic effect of CqZF-HD14 with CqNAC79 or CqHIPP34 further enhanced the drought tolerance of quinoa seedlings. Taken together, the results indicate that CqZF-HD14, CqNAC79 and CqHIPP34 may be important contributors to the drought tolerance regulatory network in quinoa, and these findings add new members to the drought tolerance gene pool.

Involvement of Auxin-Mediated CqEXPA50 Contributes to Salt Tolerance in Quinoa (Chenopodium quinoa) by Interaction with Auxin Pathway Genes.

Soil salinization is a global problem that limits crop yields and threatens agricultural development. Auxin-induced expansins contribute to plant salt tolerance through cell wall loosening. However, how auxins and expansins contribute to the adaptation of the halophyte quinoa (Chenopodium quinoa) to salt stress has not yet been reported. Here, auxin was found to contribute to the salt tolerance of quinoa by promoting the accumulation of photosynthetic pigments under salt stress, maintaining enzymatic and nonenzymatic antioxidant systems and scavenging excess reactive oxygen species (ROS). The Chenopodium quinoa expansin (Cqexpansin) family and the auxin pathway gene family (Chenopodium quinoa auxin response factor (CqARF), Chenopodium quinoa auxin/indoleacetic acid (CqAux/IAA), Chenopodium quinoa Gretchen Hagen 3 (CqGH3) and Chenopodium quinoa small auxin upregulated RNA (CqSAUR)) were identified from the quinoa genome. Combined expression profiling identified Chenopodium quinoa α-expansin 50 (CqEXPA50) as being involved in auxin-mediated salt tolerance. CqEXPA50 enhanced salt tolerance in quinoa seedlings was revealed by transient overexpression and physiological and biochemical analyses. Furthermore, the auxin pathway and salt stress-related genes regulated by CqEXPA50 were identified. The interaction of CqEXPA50 with these proteins was demonstrated by bimolecular fluorescence complementation (BIFC). The proteins that interact with CqEXPA50 were also found to improve salt tolerance. In conclusion, this study identified some genes potentially involved in the salt tolerance regulatory network of quinoa, providing new insights into salt tolerance.

Malate-mediated CqMADS68 enhances aluminum tolerance in quinoa seedlings through interaction with CqSTOP6, CqALMT6 and CqWRKY88.

Aluminum (Al) stress in acidic soils has severe negative effects on crop productivity. In this study, the alleviating effect and related mechanism of malate on Al stress in quinoa (Chenopodium quinoa) seedlings were investigated. The findings indicated that malate alleviated the growth inhibition of quinoa seedlings under Al stress, maintained the enzymatic and nonenzymatic antioxidant systems, and aided resistance to the damage caused by excessive reactive oxygen species (ROS). Under Al stress, malate significantly increased the contents of chlorophyll and carotenoids in quinoa shoots by 103.8% and 240.7%, and significantly increased the ratios of glutathione (GSH)/oxidized glutathione (GSSG), and ascorbate (AsA)/dehydroascorbate (DHA) in roots by 59.9% and 699.2%, respectively. However, malate significantly decreased the superoxide radical (O2•-), hydrogen peroxide (H2O2), malondialdehyde (MDA) and Al contents in quinoa roots under Al stress by 32.7%, 60.9%, 63.1% and 49%, respectively. Moreover, the CqMADS family and the Al stress-responsive gene families (CqSTOP, CqALMT, and CqWRKY) were identified from the quinoa genome. Comprehensive expression profiling identified CqMADS68 as being involved in malate-mediated Al resistance. Transient overexpression of CqMADS68 increased Al tolerance in quinoa seedlings. More importantly, we found that CqMADS68 regulated the expression of CqSTOP6, CqALMT6 and CqWRKY88 and further demonstrated the interaction of CqMADS68 with CqSTOP6, CqALMT6 and CqWRKY88 by bimolecular fluorescence complementation (BIFC) experiments. Moreover, transient overexpression and physiological and biochemical analyses demonstrated that CqSTOP6, CqALMT6 and CqWRKY88 could also improve Al tolerance by maintaining the antioxidant capacity of quinoa seedlings. Taken together, these findings reveal that CqMADS68, CqSTOP6, CqALMT6 and CqWRKY88 may be important contributors to the Al tolerance regulatory network in quinoa, providing new insights into Al stress resistance.

Extraction, Purification, Physicochemical Properties, and Activity of a New Polysaccharide From Cordyceps cicadae.

The polysaccharides from C. cicadae were extracted by ultrasonically-assisted enzymatic extraction (UAEE). Response surface analysis was used to determine the optimum parameters as follows: addition of enzymes, 0.71%; extraction temperature, 60°C; extraction time, 18 min; liquid-solid ratio, 46:1 (mL/g). The extraction yield of polysaccharide was 3.66 ± 0.87%. A novel polysaccharide fraction (JCH-a1) from C. cicadae was extracted and then purified by cellulose DEAE-32 and Sephadex G-100 anion exchange chromatography. The analysis results showed that the molar ratio of galactose, glucose, and mannose in JCH-a1 cells (60.7 kDa) was 0.89:1:0.39. JCH-a1 with a triple helix contains more α-glycosides and has strong thermal stability. Moreover, JCH-a1 showed strong antioxidant activity and acted as a strong inhibitor of α-glucosidase in vitro. In addition, JCH-a1 can prolong the lifespan of C. elegans. The present study might provide a basis for further study of JCH-a1 as an antioxidant and hypoglycemic food or drug.

Dragon fruit-kiwi fermented beverage: In vitro digestion, untargeted metabolome analysis and anti-aging activity in Caenorhabditis elegans.

The research on the development of dragon fruit and kiwi fruit through LAB-yeast compound fermentation is very limited, and there are few related fermentation products on the market. The purpose of this study was to evaluate the stability of the antioxidant capacity of fermented beverages (FB) through in vitro simulated digestion, to evaluate the changes in metabolites of juice after fermentation through untargeted metabolomics, and used Caenorhabditis elegans as a model to evaluate its anti-aging activity. The results showed that FB not only has good in vitro antioxidant activity, but also the total phenol content (TPC), total flavonoid content (TFC), ABTS scavenging ability, and hydroxyl radical scavenging ability of FB were significantly increased during gastric digestion and intestinal digestion. Metabolomics showed that the contents of phenols and flavonoids related to antioxidant increased after fermentation, and fermentation had a significant effect on organic acids and amino acids in FB. Finally, compared with the control group, although the original concentration of FB has a side-toxic effect on nematodes, the mean lifespan of C. elegans fed with 1.56% FB increased by 18.01%, SOD activity significantly increased by 96.16% and MDA content significantly decreased by 40.62%. FB has good antioxidant activity in vitro and in vivo, and the antioxidant activity is stable during the simulated digestion process.

Ligand based 3D-QSAR model, pharmacophore, molecular docking and ADME to identify potential fibroblast growth factor receptor 1 inhibitors.

The FGF/FGFR system may affect tumor cells and stromal microenvironment through autocrine and paracrine stimulation, thereby significantly promoting oncogene transformation and tumor growth. Abnormal expression of FGFR1 in cells is considered to be the main cause of tumorigenesis and a potential target for the treatment of cancer. In this study, a combination of structure-based drug carriers and molecular docking-based virtual screening was used to screen new potential FGFR1 inhibitors. Forty eight known inhibitors were collected to establish 3 D-QSAR models and pharmacophore models, investigate the relationship between the activity and conformation of compounds, and verify the efficiency of pharmacophore. In Accelrys Discovery Studio 2016, the ZINC database was filtered by Lipinski's Rule of Five and SMART's filtration. Then, Hypo01 was used for virtual screening of ZINC database. Compounds with predicted activity values less than 1 µM were molecularly docked with FGFR1 protein crystals, the docking results were observed, and the interaction between compounds and targets was studied. The absorption, distribution, metabolism and excretion (ADME) and toxicity of potential inhibitors were studied, and a compound with new structural scaffolds were obtained. It could be further studied to explore their better therapeutic effects. Communicated by Ramaswamy H. Sarma.

Diversity, Chemical Constituents, and Biological Activities of Endophytic Fungi Isolated From Ligusticum chuanxiong Hort.

The objective of this study was to evaluate the diversity of endophytic fungi of different parts of Ligusticum chuanxiong Hort (CX) and further characterize their biological activities and identify chemical compounds produced by these endophytic fungi. A total of 21 endophytic fungi were isolated and identified from CX. Penicillium oxalicum, Simplicillium sp., and Colletotrichum sp. were identified as promising strains by the color reaction. Comparing different organic extracts of the three strains, it was observed that the ethyl acetate extract of Penicillium oxalicum and Simplicillium sp. and the n-butanol extract of Colletotrichum sp. showed significant antioxidant and antibacterial activities. The ethyl acetate extracts of Penicillium oxalicum had outstanding antioxidant and antibacterial effects, and its radical scavenging rates for ABTS and DPPH were 98.43 ± 0.006% and 90.11 ± 0.032%, respectively. At the same time, their IC50 values were only 0.18 ± 0.02 mg/mL and 0.04 ± 0.003 mg/mL. The ethyl acetate extract of Penicillium oxalicum showed MIC value of only 0.5 mg/mL against Escherichia coli and Staphylococcus aureus. By liquid chromatography-mass spectrometry (LC-MS), we found that Penicillium oxalicum could produce many high-value polyphenols, such as hesperidin (36.06 µmol/g), ferulic acid (1.17 µmol/g), and alternariol (12.64 µmol/g), which can be a potential resource for the pharmaceutical industry. In conclusion, these results increase the diversity of CX endophytic fungi and the antioxidant and antibacterial activities of their secondary metabolites.

Therapeutic effect of gefitinib on patients with advanced EGFR-mutation NSCLC.

This study aims to study the role of gefitinib on patients with advanced EGFR-mutation NSCLC (Non-Small Cell Lung Cancer). Totally 115 patients with advanced EGFR-mutation NSCLC treated in our hospital were enrolled as research objects. They were randomly divided into control group (n=57) applied with cisplatin ± pemetrexed and experimental group (n=58) subject to gefitinib± cisplatin ± pemetrexed, both groups were applied with treatment for 4 cycles. Clinical efficacy: The disease control rate (DCR) was 72.41% in the experimental group, which was higher than that of the control group (54.39%, p<0.05); Serum CEA, CYFRA21-1, MMP-9 levels: after 2 and 4 cycles of treatment, serum CEA, CYFRA21-1, and MMP-9 levels were lower in the experimental group (p<0.05); Immune function: after 2 and 4 cycles of treatment, Th1 cells and Th1/Th2 cell levels were higher in the experimental group, while Th2 cell level was higher in the control group (p<0.05); Angiogenesis related indicators: the levels of VEGF, HIF-1α and sCD105 were lower in the experimental group after 2 and 4 cycles of treatment (p<0.05); (5) Adverse reactions: After 2 and 4 cycles of treatment, the levels of VEGF, HIF-1α, and sCD105 were lower in the experimental group (p<0.05). The application of gefitinib in patients with advanced EGFR-mutation NSCLC can help down-regulate CEA, CYFRA21-1, and MMP-9 levels, inhibit angiopoiesis, enhance immune function, and increase disease control rate.

Tartary Buckwheat (Fagopyrum tataricum) NAC Transcription Factors FtNAC16 Negatively Regulates of Pod Cracking and Salinity Tolerant in Arabidopsis.

The thick and hard fruit shell of Fagopyrum tataricum (F. tataricum) represents a processing bottleneck. At the same time, soil salinization is one of the main problems faced by modern agricultural production. Bioinformatic analysis indicated that the F. tataricum transcription factor FtNAC16 could regulate the hull cracking of F. tataricum, and the function of this transcription factor was verified by genetic transformation of Arabidopsis thaliana (A. thaliana). Phenotypic observations of the wild-type (WT), OE-FtNAC16, nst1/3 and nst1/3-FtNAC16 plant lines confirmed that FtNAC16 negatively regulated pod cracking by downregulating lignin synthesis. Under salt stress, several physiological indicators (POD, GSH, Pro and MDA) were measured, A. thaliana leaves were stained with NBT (Nitroblue Tetrazolium) and DAB (3,3'-diaminobenzidine), and all genes encoding enzymes in the lignin synthesis pathway were analyzed. These experiments confirmed that FtNAC16 increased plant sensitivity by reducing the lignin content or changing the proportions of the lignin monomer. The results of this study may help to elucidate the possible association between changes in lignin monomer synthesis and salt stress and may also contribute to fully understanding the effects of FtNAC16 on plant growth and development, particularly regarding fruit pod cracking and environmental adaptability. In future studies, it may be useful to obtain suitable cracking varieties and salt-tolerant crops through molecular breeding.

Enhanced antioxidant capacity and upregulated transporter genes contribute to the UV-B-induced increase in blinin in Conyza blinii.

Conyza blinii (C. blinii) is a traditional Chinese medicinal plant mainly grown in Sichuan, China. C. blinii is suitable for studying the mechanism of plant tolerance to UV-B due to its living conditions, characterized by a high altitude and exposure to strong ultraviolet radiation. Our results showed that the growth and photosynthetic activity of C. blinii were improved under a specific intensity of UV-B, rather than being significantly inhibited. Although UV-B increased the content of reactive oxygen species (ROS) in C. blinii, the activities of antioxidative enzymes were elevated, including superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX), which contributed to the elimination of ROS. Additionally, the content of blinin, the characteristic diterpene in C. blinii, was markedly increased by UV-B. Furthermore, RNA sequencing analyses were used to explore the molecular mechanism of UV-B tolerance in C. blinii. According to the results, most of the key enzyme genes in the blinin synthesis pathway were upregulated by UV-B. In addition, 23 upregulated terpene transporter genes were identified, and these genes might participate in blinin transport during the response to UV-B. Taken together, these results implied that enhanced antioxidant capacity and upregulated transporter genes contributed to increased synthesis of blinin in response to UV-B in C. blinii.

Improving decolorization of dyes by laccase from Bacillus licheniformis by random and site-directed mutagenesis.

BACKGROUND: Dye wastewater increases cancer risk in humans. For the treatment of dyestuffs, biodegradation has the advantages of economy, high efficiency, and environmental protection compared with traditional physical and chemical methods. Laccase is the best candidate for dye degradation because of its multiple substrates and pollution-free products. METHODS: Here, we modified the laccase gene of Bacillus licheniformis by error-prone PCR and site-directed mutagenesis and expressed in E. coli. The protein was purified by His-tagged protein purification kit. We tested the enzymatic properties of wild type and mutant laccase by single factor test, and further evaluated the decolorization ability of laccase to acid violet, alphazurine A, and methyl orange by spectrophotometry. RESULTS: Mutant laccase Lacep69and D500G were superior to wild type laccase in enzyme activity, stability, and decolorization ability. Moreover, the laccase D500G obtained by site-directed mutagenesis had higher enzyme activity in both, and the specific activity of the purified enzyme was as high as 426.13 U/mg. Also, D500G has a higher optimum temperature of 70 °C and temperature stability, while it has a more neutral pH 4.5 and pH stability. D500G had the maximum enzyme activity at a copper ion concentration of 12 mM. The results of decolorization experiments showed that D500G had a strong overall decolorization ability, with a lower decolorization rate of 18% for methyl orange and a higher decolorization rate of 78% for acid violet. CONCLUSION: Compared with the wild type laccase, the enzyme activity of D500G was significantly increased. At the same time, it has obvious advantages in the decolorization effect of different dyes. Also, the advantages of temperature and pH stability increase its tolerance to the environment of dye wastewater.