Genome-Wide Identification of Sweet Orange WRKY Transcription Factors and Analysis of Their Expression in Response to Infection by Penicillium digitatum.

WRKY transcription factors (TFs) play a vital role in plant stress signal transduction and regulate the expression of various stress resistance genes. Sweet orange (Citrus sinensis) accounts for a large proportion of the world's citrus industry, which has high economic value, while Penicillium digitatum is a prime pathogenic causing postharvest rot of oranges. There are few reports on how CsWRKY TFs play their regulatory roles after P. digitatum infects the fruit. In this study, we performed genome-wide identification, classification, phylogenetic and conserved domain analysis of CsWRKY TFs, visualized the structure and chromosomal localization of the encoded genes, explored the expression pattern of each CsWRKY gene under P. digitatum stress by transcriptome data, and made the functional prediction of the related genes. This study provided insight into the characteristics of 47 CsWRKY TFs, which were divided into three subfamilies and eight subgroups. TFs coding genes were unevenly distributed on nine chromosomes. The visualized results of the intron-exon structure and domain are closely related to phylogeny, and widely distributed cis-regulatory elements on each gene played a global regulatory role in gene expression. The expansion of the CSWRKY TFs family was probably facilitated by twenty-one pairs of duplicated genes, and the results of Ka/Ks calculations indicated that this gene family was primarily subjected to purifying selection during evolution. Our transcriptome data showed that 95.7% of WRKY genes were involved in the transcriptional regulation of sweet orange in response to P. digitatum infection. We obtained 15 differentially expressed genes and used the reported function of AtWRKY genes as references. They may be involved in defense against P. digitatum and other pathogens, closely related to the stress responses during plant growth and development. Two interesting genes, CsWRKY2 and CsWRKY14, were expressed more than 60 times and could be used as excellent candidate genes in sweet orange genetic improvement. This study offers a theoretical basis for the response of CSWRKY TFs to P. digitatum infection and provides a vital reference for molecular breeding.

Isopimaric acid, an ion channel regulator, regulates calcium and oxidative phosphorylation pathways to inhibit breast cancer proliferation and metastasis.

Breast cancer is the globally most common malignant tumor and the biggest threat to women. Even though the diagnosis and treatment of breast cancer are progressing continually, a large number of breast cancer patients eventually develop a metastatic tumor, especially triple-negative breast cancer (TNBC). Recently, metal ion homeostasis and ion signaling pathway have become important targets for cancer therapy. In this study, We analyzed the effects and mechanisms of isopimaric acid (IPA), an ion channel regulator, on the proliferation and metastasis of breast cancer cells (4 T1, MDA-MB-231and MCF-7) by cell functional assay, flow cytometry, western blot, proteomics and other techniques in vitro and in vivo. Results found that IPA significantly inhibited the proliferation and metastasis of breast cancer cells (especially 4 T1). Further studies on the anti-tumor mechanism of IPA suggested that IPA might affect EMT and Wnt signaling pathways by targeting mitochondria oxidative phosphorylation and Ca2+ signaling pathways, and then inducing breast cancer cell cycle arrest and apoptosis. Our research reveals the therapeutic value of IPA in breast cancer and provides a theoretical basis for the new treatment of breast cancer.

Z-Guggulsterone Induces Cell Cycle Arrest and Apoptosis by Targeting the p53/CCNB1/PLK1 Pathway in Triple-Negative Breast Cancer.

Myrrh is the dried resin of Commiphora Myrrh Engl., which exerts anticancer properties. However, its effects and molecular mechanisms in triple-negative breast cancer (TNBC) remain unclear. In this study, we used network pharmacology to screen Z-Guggulsterone (Z-GS) as a characteristic active component of myrrh. Cell Counting Kit-8 proliferation assays showed that Z-GS inhibited proliferation of the TNBC cell lines MDA-MB-468 and BT-549. Transwell assays also showed that Z-GS inhibited TNBC migration and invasion phenotypes. Our network pharmacology combined with RNA-sequencing analyses showed that Z-GS affected cell cycle and apoptosis processes in TNBC cells, mainly via p53 signaling, to regulate key CCNB1 (cyclin B1), PLK1 (polo-like kinase 1), and p53 targets. Flow cytometry revealed that Z-GS arrested the cell cycle at the G2/M phase and increased apoptosis in TNBC cells. Western blotting and quantitative real-time polymerase chain reaction studies confirmed that Z-GS functioned via the p53-mediated downregulation of CCNB1 and PLK1 expression. In vivo studies showed that Z-GS effectively inhibited TNBC progression. Collectively, Z-GS exhibited potential anti-TNBC activity and may functions via the p53/CCNB1/PLK1 pathway.

Bacterial second messenger c-di-GMP: Emerging functions in stress resistance.

In natural environments, bacteria constantly encounter various stressful conditions, including nutrient starvation, toxic chemicals, and oxidative stress. The ability to adapt to these adverse conditions is crucial for bacterial survival. Frequently, bacteria utilize nucleotide signaling molecules such as cyclic diguanylate (c-di-GMP) to regulate their behaviors when encounter stress conditions. c-di-GMP is a ubiquitous bacterial second messenger regulating the transition between the planktonic state and biofilm state. An essential feature of biofilms is the production of extracellular matrix that covers bacterial cells and offers a physical barrier protecting the cells from environmental assaults. Beyond that, accumulating evidences have demonstrated that changes in the environment, including stress stimuli, cause the alteration of intracellular levels of c-di-GMP in bacterial cells, which is immediately sensed by a variety of downstream effectors that induce an appropriate stress response. In this review, we summarize recent research on the role of c-di-GMP signaling in bacterial responses to diverse stress conditions.

Structural characteristics and ameliorative effect of a polysaccharide from Corbicula fluminea industrial distillate against acute liver injury induced by CCl4 in mice.

Corbicula fluminea distillate as an important industrial by-product of C. fluminea during steaming process is rich in amino acids, proteins and polysaccharides, showing potential hepatoprotective effect. In this study, a polysaccharide (CFDP) was obtained from C. fluminea distillate by three-phase partitioning combined with (NH4)2SO4 precipitation at a saturation of 60 %. The structural characteristics, antioxidant activity in vitro, and hepatoprotection against mice CCl4-induced acute liver damage of CFDP were studied. Results demonstrated that CFDP was a water-soluble homogenous polysaccharide predominantly comprising glucose (>98 %), with a weight-average molecular weight of 1.4 × 107 Da, and exhibiting potent antioxidant benefits in vitro. CFDP had a backbone of (1 â†’ 4)-α-d-glucopyranosyl (Glcp) and a small amount of (1 â†’ 4, 6)-α-D-Glcp. The branch formed at C-6 comprised by (1→)-α-D-Glcp and (1→)-α-D-N-acetylglucosamine. CFDP possessed excellent hepatoprotective activity against acute liver damage caused by CCl4 in mice, mainly by ameliorating weight reduction and organ injures, alleviating hepatic function and serum lipid metabolism, suppressing oxidative stress and inflammatory responses, as directly verified by histopathological examination. Moreover, CFDP improved gut microbiota by up-regulating the relative abundance of total bacteria and probiotics such as Firmicutes, Bacteroidete, Rumminococcaceae, Lactobacillaceae, accompanied by promoting short chain fatty acid production. Therefore, our findings indicated that CFDP can be developed as a healthy food supplement for the prevention of chemical livery injury.

Genome-wide identification, characterization, and expression profile ofNBS-LRRgene family in sweet orange (Citrussinensis).

BACKGROUND: The NBS-LRR (nucleotide-binding site-leucine-rich repeat gene) gene family, known as the plant R (resistance) gene family with the most members, plays a significant role in plant resistance to various external adversity stresses. The NBS-LRR gene family has been researched in many plant species. Citrus is one of the most vital global cash crops, the number one fruit group, and the third most traded agricultural product world wild. However, as one of the largest citrus species, a comprehensive study of the NBS-LRR gene family has not been reported on sweet oranges. METHODS: In this study, NBS-LRR genes were identified from the Citrus sinensis genome (v3.0), with a comprehensive analysis of this gene family performed, including phylogenetic analysis, gene structure, cis-acting element of a promoter, and chromosomal localization, among others. The expression pattern of NBS-LRR genes was analyzed when sweet orange fruits were infected by Penicillium digitatum, employing experimental data from our research group. It first reported the expression patterns of NBS-LRR genes under abiotic stresses, using three transcript data from NCBI (National Center for Biotechnology Information). RESULTS: In this study, 111 NBS-LRR genes were identified in the C. sinensis genome (v3.0) and classified into seven subfamilies according to their N-terminal and C-terminal domains. The phylogenetic tree results indicate that genes containing only the NBS structural domain are more ancient in the sweet orange NBS-LRR gene family. The chromosome localization results showed that 111 NBS-LRR genes were distributed unevenly on nine chromosomes, with the most genes distributed on chromosome 1. In addition, we identified a total of 18 tandem duplication gene pairs in the sweet orange NBS-LRR gene family, and based on the Ka/Ks ratio, all of the tandem duplication genes underwent purifying selection. Transcriptome data analysis showed a significant number of NBS-LRR genes expressed under biotic and abiotic stresses, and some reached significantly different levels of expression. It indicates that the NBS-LRR gene family is vital in resistance to biotic and abiotic stresses in sweet oranges. CONCLUSION: Our study provides the first comprehensive framework on the NBS-LRR family of genes, which provides a basis for further in-depth studies on the biological functions of NBS-LRR in growth, development, and response to abiotic stresses in sweet orange.

A pectic polysaccharide from fresh okra (Abelmoschus esculentus L.) beneficially ameliorates CCl4-induced acute liver injury in mice by antioxidation, inhibition of inflammation and modulation of gut microbiota.

Okra [Abelmoschus esculentus (Linn.) Moench], as a well-known medicinal and food plant, has important physiological activities and health benefits, and polysaccharide is its main bioactive component. In this study, a pectic polysaccharide (OPS-50) prepared from fresh okra pods by three-phase partitioning and gradient (NH4)2SO4 precipitation at a saturation of 50% was employed in carbon tetrachloride (CCl4)-caused acute liver damage in mice to evaluate the hepatoprotective potential. Results indicated that OPS-50 was mainly composed of a limited linear homogalacturonan backbone and abundant rhamnogalacturonan-I domains as side chains. OPS-50 exerted positively protective effects on acute liver damage induced by CCl4 in mice through relieving weight reduction and organ damage, ameliorating liver function and dyslipidemia, alleviating oxidative stress, suppressing pro-inflammatory cytokines, modulating gut microbiota, and promoting short-chain fatty acid secretion. Moreover, liver histopathology demonstrated the protective benefit of OPS-50 on CCl4-caused acute liver damage in mice. Therefore, our data suggested that the pectic OPS-50, as a dietary supplement, have great potential in preventing and treating chemical liver damages.

All-trans retinoic acid enhances the cytotoxic effect of decitabine on myelodysplastic syndromes and acute myeloid leukaemia by activating the RARα-Nrf2 complex.

BACKGROUND: Decitabine (DAC) is used as the first-line therapy in patients with higher-risk myelodysplastic syndromes (HR-MDS) and elderly acute myeloid leukaemia (AML) patients unsuitable for intensive chemotherapy. However, the clinical outcomes of patients treated with DAC as a monotherapy are far from satisfactory. Adding all-trans retinoic acid (ATRA) to DAC reportedly benefitted MDS and elderly AML patients. However, the underlying mechanisms remain unclear and need further explorations from laboratory experiments. METHODS: We used MDS and AML cell lines and primary cells to evaluate the combined effects of DAC and ATRA as well as the underlying mechanisms. We used the MOLM-13-luciferase murine xenograft model to verify the enhanced cytotoxic effect of the drug combination. RESULTS: The combination treatment reduced the viability of MDS/AML cells in vitro, delayed leukaemia progress, and extended survival in murine xenograft models compared to non- and mono-drug treated models. DAC application as a single agent induced Nrf2 activation and downstream antioxidative response, and restrained reactive oxygen species (ROS) generation, thus leading to DAC resistance. The addition of ATRA blocked Nrf2 activation by activating the RARα-Nrf2 complex, leading to ROS accumulation and ROS-dependent cytotoxicity. CONCLUSIONS: These results demonstrate that combining DAC and ATRA has potential for the clinical treatment of HR-MDS/AML and merits further exploration.

A secreted effector with a dual role as a toxin and as a transcriptional factor.

Bacteria have evolved multiple secretion systems for delivering effector proteins into the cytosol of neighboring cells, but the roles of many of these effectors remain unknown. Here, we show that Yersinia pseudotuberculosis secretes an effector, CccR, that can act both as a toxin and as a transcriptional factor. The effector is secreted by a type VI secretion system (T6SS) and can enter nearby cells of the same species and other species (such as Escherichia coli) via cell-cell contact and in a contact-independent manner. CccR contains an N-terminal FIC domain and a C-terminal DNA-binding domain. In Y. pseudotuberculosis cells, CccR inhibits its own expression by binding through its DNA-binding domain to the cccR promoter, and affects the expression of other genes through unclear mechanisms. In E. coli cells, the FIC domain of CccR AMPylates the cell division protein FtsZ, inducing cell filamentation and growth arrest. Thus, our results indicate that CccR has a dual role, modulating gene expression in neighboring cells of the same species, and inhibiting the growth of competitors.

Transcriptome Analysis of Low-Temperature-Treated Tetraploid Yellow Actinidia chinensis Planch. Tissue Culture Plantlets.

The cold-resistant mechanism of yellow kiwifruit associated with gene regulation is poorly investigated. In this study, to provide insight into the causes of differences in low-temperature tolerance and to better understand cold-adaptive mechanisms, we treated yellow tetraploid kiwifruit 'SWFU03' tissue culture plantlets at low temperatures, used these plantlets for transcriptome analysis, and validated the expression levels of ten selected genes by real-time quantitative polymerase chain reaction (RT-qPCR) analysis. A number of 1630 differentially expressed genes (DEGs) were identified, of which 619 pathway genes were up-regulated, and 1011 were down-regulated in the cold treatment group. The DEGs enriched in the cold tolerance-related pathways mainly included the plant hormone signal transduction and the starch and sucrose metabolism pathway. RT-qPCR analysis confirmed the expression levels of eight up-regulated genes in these pathways in the cold-resistant mutants. In this study, cold tolerance-related pathways (the plant hormone signal transduction and starch and sucrose metabolism pathway) and genes, e.g., CEY00_Acc03316 (abscisic acid receptor PYL), CEY00_Acc13130 (bZIP transcription factor), CEY00_Acc33627 (TIFY protein), CEY00_Acc26744 (alpha-trehalose-phosphate synthase), CEY00_Acc28966 (beta-amylase), CEY00_Acc16756 (trehalose phosphatase), and CEY00_Acc08918 (beta-amylase 4) were found.

Analysis of RPGR gene mutations in 41 Chinese families affected by X-linked inherited retinal dystrophy.

Background: This study analyzed the phenotypes and genotypes of 41 Chinese families with inherited retinal dystrophy (IRD) and RPGR gene mutations. Methods: This retrospective analysis evaluated a cohort of 41 patients who were subjected to a specific Hereditary Eye Disease Enrichment Panel (HEDEP) analysis. All (likely) pathogenic variants were determined by Sanger sequencing, and co-segregation analyses were performed on the available family members. All cases were subjected to Sanger sequencing for RPGR open reading frame 15 (ORF15) mutations. Results: A total of 41 probands from different families with a clinical diagnosis of retinitis pigmentosa (RP; 34 cases) and cone-rod dystrophy (CORD; 7 cases) were included in this cohort. According to clinical information, 2, 18, and 21 cases were first assigned as autosomal dominant (AD), sporadic, and X-linked (XL) inheritance, respectively. Several cases of affected females who presented with a male phenotype have been described, posing challenges at diagnosis related to the apparent family history of AD. Mutations were located in RPGR exons or introns 1-14 and in ORF15 of 12 of 41 (29.3%) and 29 of 41 (70.7%) subjects, respectively. Thirty-four (likely) pathogenic mutations were identified. Frameshifts were the most frequently observed variants, followed by nonsense, splice, and missense mutations. Herein, a detailed description of four RP patients carrying RPGR intronic mutations is reported, and in vitro splice assays were performed to confirm the pathogenicity of these intronic mutations. Conclusion: Our findings provide useful insights for the genetic and clinical counseling of patients with XL IRD, which will be useful for ongoing and future gene therapy trials.

Exploring AI-2-mediated interspecies communications within rumen microbial communities.

BACKGROUND: The rumen is an ecosystem with a complex microbial microflora in which microbes initiate biofilm formation by attaching to plant surfaces for plant degradation and are capable of converting feed to nutrients and energy via microbial processes. Quorum sensing (QS) is a cell-to-cell communication mechanism that allows microbes to synchronize the expression of multiple genes in the group to perform social behaviors such as chemotaxis and biofilm formation using self-synthesized QS signaling molecules. Whereas QS has been extensively studied in model microorganisms under pure culture conditions, QS mechanisms are poorly understood in complex bacterial communities, such as the rumen microflora, in which cell-to-cell communication may be common. RESULTS: Here, we analyzed 981 rumens bacterial and archaeal genomes from the Joint Genome Institute (JGI) and GenBank databases and identified 15 types of known QS signaling molecule-related genes. The analysis of the prevalence and abundance of genes involved in QS showed that 767 microbial genomes appeared to possess QS-related genes, including 680 bacterial genomes containing autoinducer-2 (AI-2) synthase- or receptor-encoding genes. Prevotella, Butyivibrio, Ruminococcus, Oribacterium, Selenomonas, and Treponema, known abundant bacterial genera in the rumen, possessed the greatest numbers of AI-2-related genes; these genes were highly expressed within the metatranscriptome dataset, suggesting that intra- and interspecies communication mediated by AI-2 among rumen microbes was universal in the rumen. The QS processes mediated by the dCache_1-containing AI-2 receptors (CahRs) with various functional modules may be essential for degrading plants, digesting food, and providing energy and nutrients to the host. Additionally, a universal natural network based on QS revealed how rumen microbes coordinate social behaviors via the AI-2-mediated QS system, most of which may potentially function via AI-2 binding to the extracellular sensor dCache_1 domain to activate corresponding receptors involved in different signal transduction pathways, such as methyl-accepting chemotaxis proteins, histidine kinases, serine phosphatases, c-di-GMP synthases and phosphodiesterases, and serine/threonine kinases in the rumen. CONCLUSIONS: The exploration of AI-2-related genes, especially CahR-type AI-2 receptors, greatly increased our insight into AI-2 as a potentially "universal" signal mediating social behaviors and will help us better understand microbial communication networks and the function of QS in plant-microbe interactions in complex microecosystems. Video Abstract.

Recent advances in the bioactive polysaccharides and other key components from Phellinus spp. and their pharmacological effects: A review.

Phellinus spp. is one of the largest genera of Hymenochaetaceae with approximately 220 species, such as P. vaninii, P. buamii, P. linteus, and P. igniarius, these species are considered as precious food supplements and medicinal ingredients in China, Korea, Japan, and other Asian countries for over 2000 years. Phellinus spp. contains abundant bioactive polysaccharides and other key components (e.g., phenolics, terpenes, steroids, etc.). Pharmacological investigations have confirmed that bioactive polysaccharides and other important secondary metabolites from Phellinus spp. possess multiple health-promoting benefits, including antitumor, immunomodulatory, anti-inflammatory, antidiabetic, antioxidant, and antimicrobial effects. However, comprehensive evaluations on the preparation and structural characteristics, bioactivities, and toxicology of these functional components (e.g., polysaccharides, phenolics, terpenes, steroids) from various Phellinus spp. species are very limited, which may restrict the practical application of Phellinus spp. This review summarizes the physicochemical characteristics, pharmacological activities, and possible mechanisms of bioactive components from Phellinus spp. according to published studies from 2017 to 2022. It also surveyed the toxicological assessment for safety and applications of different Phellinus spp. species. This review aims to provide useful references and promising directions for the comprehensive development and utilization of Phellinus spp. in functional foods, pharmaceuticals, and cosmetics.

Experimental Study on Mechanical Properties of Rebar in Steel Half-Grouted Sleeve Connections with Construction Defects.

A prefabricated concrete structure is a building structure designed for sustainability and low comprehensive carbon emission. The grouted sleeve splice is a major connection method for prefabricated concrete structures. However, construction defects occur easily in the grouted sleeve splice connection at construction sites because of complex construction environments and the high connection accuracy. To determine the influence of rebar in steel half-grouted sleeve connections with construction defects, investigations were conducted using four different test groups (rebar offset, rebar bended, insufficient fluidity of grout, and control group). The load-displacement curve and load-stress curve were analyzed on 24 different specimens through uniaxial tension experiments. The experimental results showed that rebar fracture was the failure of specimens. The load-displacement curves consisted of elastic, yield, strength, and tight stages. The curves were similar to rebar under uniaxial tension, except for the rebar bended group. The axial stress and circumferential stress on the sleeve surface consistently followed a linear response before the specimen yield, whereas the axial stress and circumferential stress showed a rebound response after the specimen yielded. Different finite element models were established based on the different defects. Compared with the experimental results, the finite element analysis results coincided with those of the experimental results, and the errors were within 8% to evaluate the performance of steel half-grouted sleeve connections in construction.

Fungal dynamic changes in naturally fermented 'Kyoho' grape juice.

The 'Kyoho' grape (Vitaceae, Plantae) has large ears, plenty of flesh, and rich nutrition and is planted across a large area in China. There are few reports on this variety in winemaking, especially on the dynamic changes of fungi in the wine fermentation broth. In this study, we used the 'Kyoho' grapes as raw materials and adopted a high throughput to analyze dynamic changes in fungal species composition of the natural fermentation broth at four time points: day 1 (D1P), day 3 (D3P), day 5 (D5P), and day 15 (D15P). Changes in fungal metabolic pathways and dominant yeasts were also analyzed. A total of 78 families, 110 genera, and 137 species were detected, in the natural fermentation broth samples. Forty-nine families, 60 genera, and 72 species were found in the control check (CK). A total of 66 differential metabolic pathways were enriched; of those, 41 were up-regulated compared to CK, such as CDP-diacylglycerol biosynthesis I (PWY 5667), chitin degradation to ethanol (PWY 7118), and the super pathway of phosphatidate biosynthesis (PWY 7411). Changes in fungal metabolic pathways were in line with the dynamic changes of dominant yeast species in the whole process of fermentation. Pichia kluyveri, P. membranifaciens, and Citeromyces matritensis are the dominant species in the later stages of natural fermentation. These yeast species may play vital roles in the 'Kyoho' wine industry in the future.

AAV-Mediated Gene Replacement Therapy Restores Viability of BCD Patient iPSC Derived RPE Cells and Vision of Cyp4v3 Knockout Mice.

Bietti crystalline corneoretinal dystrophy (BCD) is an autosomal recessive retinal degenerative disease characterized by yellow-white crystal deposits in the posterior pole, degeneration of the retinal pigment epithelium (RPE), and sclerosis of the choroid. Mutations in the cytochrome P450 4 V2 gene (CYP4V2) cause BCD, which is associated with lipid metabolic disruption. The use of gene replacement therapy in BCD has been hampered by the lack of disease models. To advance CYP4V2 gene replacement therapy, we generated BCD patient-specific induced pluripotent stem cell (iPSC)-RPE cells and Cyp4v3 knockout (KO) mice as disease models and AAV2/8-CAG-CYP4V2 as treatment vectors. We demonstrated that after AAV-mediated CYP4V2 gene replacement therapy BCD-iPSC-RPE cells presented restored cell survival and reduced lipid droplets accumulation; restoration of vision in Cyp4v3 KO mice was revealed by elevated electroretinogram amplitude and ameliorated RPE degeneration. These results suggest that AAV-mediated gene replacement therapy in BCD patients is a promising strategy.

Mediastinal Nodal Staging Performance of Combined Endobronchial and Esophageal Endosonography in Lung Cancer Cases: A Systematic Review and Meta-Analysis.

By searching lliteratures till January 5, 2022, we evaluated the role of the mediastinal nodal staging of endobronchial ultrasound-guided fine-needle aspiration (EBUS) and endoscopic ultrasound-guided fine-needle aspiration (EUS) in lung cancer. A total of 20 studies with 2,961 patients were included in this study. The pooled sensitivity, specificity, PLR, and NLR for EBUS were 0.79, 0.97, 27.29, and 0.25, respectively. EUS showed staging performance similar to EBUS. The staging performance was significantly improved when combining EBUS + EUS.

Degradation of pesticide wastewater with simultaneous resource recovery via ozonation coupled with anaerobic biochemical technology.

The effective treatment of pesticide wastewater with high organic content, complex composition and high-toxicity has attracted enormous attention of researchers. This work proposes a new idea for removing the pesticide wastewater with simultaneous resource recovery, which is different from the traditional view of mineralization of pesticide wastewater via composite technology. This novel strategy involved a sequential three-step treatment: (a) acidic Ozonation process, to remove the venomous aromatic heterocyclic compounds; (b) hydrolysis and ozonation in alkaline conditions, enhancing the biodegradability of pesticide wastewater, mainly due to the dehalogenation, elimination of C=C bonds and production of low molecular-weight carboxylate anions; (c) the final step is anaerobic biological reactions. Based on the characterizations, this two-stage acidic-alkaline ozonation can efficiently degraded the virulence of pesticide wastewater and enhance its biodegradability from 0.08 to 0.32. The final anaerobic biochemical treatment can stably remove the residuals and convert the low molecular-weight organics into CH4, achieving the resource recovery. This work explored the pH-dependent of ozonized degradation of pesticide wastewater and gives a new perspective of wastewater treatment.

Analysis of fungal dynamic changes in the natural fermentation broth of 'Hongyang' kiwifruit.

'Hongyang' kiwifruit (Actinidia chinensis Planch.) is an ideal kiwifruit wine variety. At present, there is no research on the dynamic changes of yeast during the natural fermentation of kiwifruit wine. In this study, a high-throughput was employed to analyze the fungal population composition and diversity in the samples cultured in yeast extract peptone dextrose (YPD) medium and enriched in the natural fermentation process of 'Hongyang' kiwifruit at four time points, day one (D1T), day three (D3T), day five (D5T), and day fifteen (D15T). Five hundred and eighty-two operational taxonomic units (OTUs) were obtained from 131 genera and 178 species samples. The diversity analysis results showed that in the early natural fermentation stage, the dominant species was Aureobasidium pullulans, and as natural fermentation proceeded, the genus Pichia became the dominant species. Pichia kluyveri was an important species at the later stages of natural fermentation. An analysis of the metabolic pathways shows that P. kluyveri plays an aromatic-producing role in the natural fermentation of 'Hongyang' kiwifruit. These results could provide a theoretical basis for the studies of kiwifruit fungal diversity and fungal changes during fermentation. The findings could fix a major deficiency in the production of kiwifruit fruit wine, which lacks a specific flavor-producing yeast species or strain.

4-[1-Ethyl-1-methylhexy]-phenol induces apoptosis and interrupts Ca2+ homeostasis via ROS pathway in Sertoli TM4 cells.

Biological effect of an individual nonylphenol (NP) isomer extremely relies upon the side chain structure. This research was designed to evaluate the impact of NP isomer, 4-[1-ethyl-1-methylhexy]-phenol (NP65), on Sertoli cells in vitro. Sertoli TM4 cells were exposed to various concentration (0, 0.1, 1, 10, or 20 µM) of NP65 for 24 h, and the outcomes indicated that treatment of NP65 induced reactive oxygen species (ROS) generation, oxidative stress, and apoptosis for Sertoli TM4 cells. In addition, it was found that NP65 exposure affected homeostasis of Ca2+ in Sertoli TM4 cells by increasing cytoplasm [Ca2+]i, inhibiting Ca2+-ATPase activity and decreasing cyclic adenosine monophosphate (cAMP) concentration. Pretreatment with ROS scavenger, N-acetylcysteine (NAC), attenuated NP65-induced oxidative stress as well as apoptosis for TM4 cells. Furthermore, NAC blocked NP65-induced disorders of Ca2+ homeostasis by attenuating the growth of intracellular [Ca2+]i and the inhibition of Ca2+-ATPase and cAMP activities. Thus, we have demonstrated that NP65 induced apoptosis as well as acted as a potent inhibitor of Ca2+-ATPase activity and resulted in disorder of Ca2+ homeostasis in Sertoli TM4 cells; ROS participated in the process. Our results supported the view that oxidative stress acted an essential role within the development of apoptosis and Ca2+ overload in TM4 cells as a consequence of NP65 stimulation.