In situ profiling reveals spatially metabolic injury in the initiation of polystyrene nanoplastic-derived intestinal epithelial injury in mice.

Despite increasing concerns regarding the harmful effects of plastic-induced gut injury, mechanisms underlying the initiation of plastic-derived intestinal toxicity remain unelucidated. Here, mice were subjected to long-term exposure to polystyrene nanoplastics (PS-NPs) of varying sizes (80, 200, and 1000 nm) at doses relevant to human dietary exposure. PS-NPs exposure did not induce a significant inflammatory response, histopathological damage, or intestinal epithelial dysfunction in mice at a dosage of 0.5 mg/kg/day for 28 days. However, PS-NPs were detected in the mouse intestine, coupled with observed microstructural changes in enterocytes, including mild villous lodging, mitochondrial membrane rupture, and endoplasmic reticulum (ER) dysfunction, suggesting that intestinal-accumulating PS-NPs resulted in the onset of intestinal epithelial injury in mice. Mechanistically, intragastric PS-NPs induced gut microbiota dysbiosis and specific bacteria alterations, accompanied by abnormal metabolic fingerprinting in the plasma. Furthermore, integrated data from mass spectrometry imaging-based spatial metabolomics and metallomics revealed that PS-NPs exposure led to gut dysbiosis-associated host metabolic reprogramming and initiated intestinal injury. These findings provide novel insights into the critical gut microbial-host metabolic remodeling events vital to nanoplastic-derived-initiated intestinal injury.

Gene identification and semisynthesis of the anti-inflammatory oleanane-type triterpenoid wilforlide A.

Wilforlide A is one of the main active constituents produced in trace amounts in Tripterygium wilfordii Hook F, which has excellent anti-inflammatory and immune suppressive effects. Despite the seeming structural simplicity of the compound, the biosynthetic pathway of wilforlide A remains unknown. Gene-specific expression analysis and genome mining were used to identify the gene candidates, and their functions were studied in vitro and in vivo. A modularized two-step (M2S) technique and CRISPR-Cas9 methods were used to construct engineering yeast. Here, we identified a cytochrome P450, TwCYP82AS1, that catalyses C-22 hydroxylation during wilforlide A biosynthesis. We also found that TwCYP712K1 to K3 can further oxidize the C-29 carboxylation of oleanane-type triterpenes in addition to friedelane-type triterpenes. Reconstitution of the biosynthetic pathway in engineered yeast increased the precursor supply, and combining TwCYP82AS1 and TwCYP712Ks produced abrusgenic acid, which was briefly acidified to achieve the semisynthesis of wilforlide A. Our work presents an alternative metabolic engineering approach for obtaining wilforlide A without relying on extraction from plants.

Crucial roles of sorbitol metabolism and energy status in the chilling tolerance of yellow peach.

In this study, we compared sorbitol metabolism, energy metabolism, and CI development in yellow peach fruit at 1 °C (less susceptible to CI) and 8 °C (more susceptible to CI) storage to elucidate potential connections between them. The results indicated that storage at 1 °C effectively maintained the textural quality of yellow peach fruit and delayed the onset of CI by 12 days compared to 8 °C. This positive effect might be attributable to 1 °C storage maintaining higher sorbitol content throughout the storage duration, thus sustaining the higher adenosine triphosphate (ATP) level and energy charge. The regulation of sorbitol accumulation by 1 °C storage was closely linked to the metabolic activity of sorbitol, which stimulated sorbitol synthesis by enhancing sorbitol-6-phosphate dehydrogenase (S6PDH) activity after 12 days while suppressing sorbitol degradation via decreased sorbitol oxidase (SOX) and NAD+-sorbitol dehydrogenase (NAD+-SDH) activities before 24 days. In addition, the notable up-regulation in the NAD+-SDH activity in the late storage period promoted the conversion of sorbitol to fructose and glucose under 1 °C storage, thereby providing ample energy substrate for ATP generation. Moreover, sorbitol acts as a vital signaling molecule, and substantially up-regulated expressions of sorbitol transporters genes (PpeSOT3, PpeSOT5, and PpeSOT7) were observed in fruit stored at 1 °C, which might promote sorbitol transport and improve cold tolerance in peach fruit. Taken together, these findings suggested that 1 °C storage delayed CI by enhancing sorbitol metabolism and transporter activity, promoting sorbitol accumulation, and finally elevating the energy status in yellow peach fruit.

CYP72D19 from Tripterygium wilfordii catalyzes C-2 hydroxylation of abietane-type diterpenoids.

KEY MESSAGE: CYP72D19, the first functional gene of the CYP72D subfamily, catalyzes the C-2 hydroxylation of abietane-type diterpenoids. The abietane-type diterpenoids, e.g., triptolide, tripdiolide, and 2-epitripdiolide, are the main natural products for the anti-tumor, anti-inflammatory, and immunosuppressive activities of Tripterygium wilfordii, while their biosynthetic pathways are not resolved. Here, we cloned and characterized the CYP72D19-catalyzed C-2 hydroxylation of dehydroabietic acid, a compound that has been proven to be a biosynthetic intermediate in triptolide biosynthesis. Through molecular docking and site-directed mutagenesis, L386, L387, and I493 near the active pocket were found to have an important effect on the enzyme activity, which also indicates that steric hindrance of residues plays an important role in function. In addition, CYP72D19 also catalyzed a variety of abietane-type diterpenoids with benzene ring, presumably because the benzene ring of the substrate molecule stabilized the C-ring, allowing the protein and the substrate to form a relatively stable spatial structure. This is the first demonstration of CYP72D subfamily gene function. Our research provides important genetic elements for the structural modification of active ingredients and the heterologous production of other 2-hydroxyl abietane-type natural products.

Multiple-statistical genome-wide association analysis and genomic prediction of fruit aroma and agronomic traits in peaches.

'Chinese Cling' is an important founder in peach breeding history due to the pleasant flavor. Genome-wide association studies (GWAS) combined with genomic selection are promising tools in fruit tree breeding, as there is a considerable time lapse between crossing and release of a cultivar. In this study, 242 peaches from Shanghai germplasm were genotyped with 145 456 single-nucleotide polymorphisms (SNPs). The six agronomic traits of fruit flesh color, fruit shape, fruit hairiness, flower type, pollen sterility, and soluble solids content, along with 14 key volatile odor compounds (VOCs), were recorded for multiple-statistical GWAS. Except the reported candidate genes, six novel genes were identified as associated with these traits. Thirty-nine significant SNPs were associated with eight VOCs. The putative candidate genes were confirmed for VOCs by RNA-seq, including three genes in the biosynthesis pathway found to be associated with linalool, soluble solids content, and cis-3-hexenyl acetate. Multiple-trait genomic prediction enhanced the predictive ability for γ-decalactone to 0.7415 compared with the single-trait model value of 0.1017. One PTS1-SSR marker was designed to predict the linalool content, and the favorable genotype 187/187 was confirmed, mainly existing in the 'Shanghai Shuimi' landrace. Overall, our findings will be helpful in determining peach accessions with the ideal phenotype and show the potential of multiple-trait genomic prediction to improve accuracy for highly correlated genetic traits. The diagnostic marker will be valuable for the breeder to bridge the gap between quantitative trait loci and marker-assisted selection for developing strong-aroma cultivars.

[Gene clone and functional identification of sterol glycosyltransferases from Paris polyphylla var. yunnanensis].

In this study, the authors cloned a glycosyltransferase gene PpUGT2 from Paris polyphylla var. yunnanensis with the ORF length of 1 773 bp and encoding 590 amino acids. The phylogenetic tree revealed that PpUGT2 belonged to the UGT80A subfamily and was named as UGT80A49 by the UDP-glycosyltransferase(UGT) Nomenclature Committee. The expression vector pET28a-PpUGT2 was constructed, and enzyme catalytic reaction in vitro was conducted via inducing protein expression and extraction. With UDP-glucose as sugar donor and diosgenin and pennogenin as substrates, the protein was found with the ability to catalyze the C-3 hydroxyl ß-glycosylation of diosgenin and pennogenin. To further explore its catalytic characteristic, 15 substrates including steroids and triterpenes were selected and PpUGT2 showed its activity towards the C-17 position of sterol testosterone with UDP-glucose as sugar donor. Homology modelling and molecule docking of PpUGT2 with substrates predicted the key residues interacting with ligands. The re-levant residues of PpUGT2-ligand binding model were scanned to calculate the corresponding mutants, and the optimized mutants were obtained according to the changes in binding affinity of the ligand with protein and the surrounding residues within 5.0 Å of ligands, which had reference value for design of the mutants. This study laid a foundation for further exploring the biosynthetic pathway of polyphyllin as well as the structure of sterol glycosyltransferases.

Transcriptome Co-Expression Network Analysis of Peach Fruit with Different Sugar Concentrations Reveals Key Regulators in Sugar Metabolism Involved in Cold Tolerance.

Peach fruits are known to be highly susceptible to chilling injury (CI) during low-temperature storage, which has been linked to the level of sugar concentration in the fruit. In order to better understand the relationship between sugar metabolism and CI, we conducted a study examining the concentration of sucrose, fructose, and glucose in peach fruit with different sugar concentrations and examined their relationship with CI. Through transcriptome sequencing, we screened the functional genes and transcription factors (TFs) involved in the sugar metabolism pathway that may cause CI in peach fruit. Our results identified five key functional genes (PpSS, PpINV, PpMGAM, PpFRK, and PpHXK) and eight TFs (PpMYB1/3, PpMYB-related1, PpWRKY4, PpbZIP1/2/3, and PpbHLH2) that are associated with sugar metabolism and CI development. The analysis of co-expression network mapping and binding site prediction identified the most likely associations between these TFs and functional genes. This study provides insights into the metabolic and molecular mechanisms regulating sugar changes in peach fruit with different sugar concentrations and presents potential targets for breeding high-sugar and cold-tolerant peach varieties.

ß-adrenoreceptor-triggered PKA activation negatively regulates the innate antiviral response.

Upon viral infection, cytoplasmic pattern recognition receptors detect viral nucleic acids and activate the adaptor protein VISA/MAVS- or MITA/STING-mediated innate antiviral response. Whether and how the innate antiviral response is regulated by neuronal endocrine functions is unclear. Here, we show that viral infection reduced the serum levels of the ß-adrenergic hormones epinephrine and norepinephrine as well as the cellular levels of their receptors ADRB1 and ADRB2. We further show that an increase in epinephrine/norepinephrine level inhibited the innate antiviral response in an ADRB1-/2-dependent manner. Mechanistically, epinephrine/norepinephrine stimulation activated the downstream kinase PKA, which catalyzed the phosphorylation of MITA at S241, S243 and T263, inhibiting MITA activation and suppressing the innate immune response to DNA virus. In addition, phosphorylation of VISA at T54 by PKA antagonized the innate immune response to RNA virus. These findings reveal the regulatory mechanisms of innate antiviral responses by epinephrine/norepinephrine and provide a possible explanation for increased host susceptibility to viral infection in stressful and anxiety-promoting situations.

A novel sterol glycosyltransferase catalyses steroidal sapogenin 3-O glucosylation from Paris polyphylla var. yunnanensis.

BACKGROUND: Paris polyphylla var. yunnanensis is an important medicinal plant, and the main active ingredient of the plant is polyphyllin, which is a steroid saponin with pharmacological activities. The central enzyme genes participating in the biosynthesis of polyphyllin are increasingly being uncovered; however, UGTs are rarely illustrated. METHODS AND RESULTS: In this study, we cloned a new sterol glycosyltransferase from Paris polyphylla var. yunnanensis and identified its catalytic function in vitro. PpUGT6 showed the ability to catalyse the C-3 glycosylation of pennogenin sapogenin of polyphyllin, and PpUGT6 showed catalytic promiscuity towards steroids at the C-17 position of testosterone and methyltestosterone and the triterpene at the C-3 position of glycyrrhetinic acid. Homology modelling of the PpUGT6 protein and virtual molecular docking of PpUGT6 with sugar acceptors and donors were performed, and we predicted the key residues interacting with ligands. CONCLUSIONS: Here, PpUGT6, a novel sterol glycosyltransferase related to the biosynthesis of polyphyllin from P. polyphylla, was characterized. PpUGT6 catalysed C-3 glycosylation to pennogenin sapogenin of polyphyllin, which is the first glycosylation step of the biosynthetic pathway of polyphyllins. Interestingly, PpUGT6 demonstrated glycodiversification to testosterone and methyltestosterone at C-17 and triterpene of glycyrrhetinic acid at the C-3 position. The virtual molecular docking of PpUGT6 protein with ligands predicted the key residues interacting with them. This work characterized a novel SGT glycosylating pennogenin sapogenin at C-3 of polyphyllin from P. polyphylla and provided a reference for further elucidation of the phytosterol glycosyltransferases in catalytic promiscuity and key residues interacting with substrates.

Profiling of naturally occurring proanthocyanidins and other phenolic compounds in a diverse peach germplasm by LC-MS/MS.

Peach fruits are rich in phenolic compounds and have considerable health benefits. In this study, 19 proanthocyanidins (PACs) and 37 other phenolic compounds are identified and evaluated in the mature fruits of 217 peach accessions using LC-QTRAP-MS/MS and LC-QTOF-MS analyses. Total PAC quantities ranged from 18.93 to 697.52 µg/g fresh weight with a variance of 36.8-fold, and accounted for 11.2 % - 85.6 % of total phenolics content. Kaempferol-3-O-rutinoside (0.001-0.968 µg/g), isorhamnetin-3-O-rutinoside (0.001-0.300 µg/g), taxifolin (0.006-0.078 µg/g), luteoloside (0.002-0.068 µg/g), prunin (0.043-33.333 µg/g), phlorizin (0.018-1.100 µg/g), and trans-piceid (0.013-0.472 µg/g) were also highly diverse. The fruit ripening period, breeding background and fruit type significantly influenced the PACs and phenolic glycosides. This study presents a complete profile of PACs and other major phenolics in 217 peach germplasms, and is expected to aid future peach breeding procedures targeted at producing plants rich in specific phenolics.

Genome-wide analysis of terpene synthase gene family to explore candidate genes related to disease resistance in Prunus persica.

In plants, a family of terpene synthases (TPSs) is responsible for the biosynthesis of terpenes and contributes to species-specific diversity of volatile organic compounds, which play essential roles in fitness of plants. However, little is known about the TPS gene family in peach and/or nectarine (Prunus persica L.). In this study, we identified 40 PpTPS genes in peach genome v2.0. Although these PpTPSs could be clustered into five classes, they distribute in several gene clusters of three chromosomes, share conserved exon-intron organizations, and code similar protein motifs. Thirty-five PpTPSs, especially PpTPS2, PpTPS23, PpTPS17, PpTPS18, and PpTPS19, altered their transcript levels after inoculation with Botryosphaeria dothidea, a cause of peach gummosis, compared to the mock treatments, which might further affect the contents of 133 terpenoids at 48 hours and/or 84 hours post inoculations in the current-year shoots of 'Huyou018', a highly susceptible nectarine cultivar. Moreover, about fifteen PpTPSs, such as PpTPS1, PpTPS2, PpTPS3, and PpTPS5, showed distinct expression patterns during fruit development and ripening in two peach cultivars, yellow-fleshed 'Jinchun' and white-fleshed 'Hikawa Hakuho'. Among them, the transcription level of chloroplast-localized PpTPS3 was obviously related to the content of linalool in fruit pulps. In addition, elevated concentrations (0.1 g/L to 1.0 g/L) of linalool showed antifungal activities in PDA medium. These results improve our understanding of peach PpTPS genes and their potential roles in defense responses against pathogens.

Controlled atmosphere storage alleviates internal browning in flat peach fruit by regulating energy and sugar metabolisms.

Flat peach fruit are cold-sensitive and vulnerable to chilling injury (CI), particularly internal browning (IB) during cold storage, which limits the consumer acceptance and market value of the fruit. Controlled atmosphere (CA) has been used to alleviate IB in fruit. However, the mechanisms of CA on IB in peach remains unknown. This study investigated the effects of CA (3-3.5% Oxygen, 3-3.5% Carbon dioxide, and 93-94% nitrogen) treatment on IB development, sugar metabolism, and energy metabolism in cold-stored (1 ± 0.5 °C) peach. The CA treatment effectively inhibited the development of IB and markedly inhibited the reduction of sugar contents and energy charge. The protein expression of the V-type proton ATPase subunit was significantly inhibited by the CA treatment, accompanied by higher adenosine triphosphate (ATP) content, and energy charge than the control fruit. Notably, the expressions of the pyruvate kinase family of proteins, pyruvate decarboxylases, and sucrose synthase were induced by CA treatment that had complex protein interactions with the ATPase and the energy metabolism pathway. These results indicated that CA treatment enhanced the chilling tolerance attributed to maintaining higher levels of energy status and sugar contents by regulating the expression of key proteins involved in energy metabolism during cold storage and shelf life. Taken together, our study can provide theoretical support for the research and development of fresh-keeping and cold-chain logistics technology.

Post-veraison different frequencies of water deficit strategies enhance Reliance grapes quality under root restriction.

In this study, two water deficit treatments in the same amount of water but with different frequencies (T1: 2.5 L per 4 d and T2: 5 L per 8 d) were performed on Reliance grapevines from veraison until harvest to explore their effects on grape berries quality under root restriction. Results showed that glucose, fructose and sucrose contents were increased, while malic acid, tartaric acid and citric acid contents were decreased under two treatments. Meanwhile, water deficits also promoted the accumulation of phenylalanine and proline. For phenols, anthocyanins, resveratrol and flavonols contents in the water deficit groups were significantly higher than those in the control group. In addition, two water deficit treatments increased the characteristic aromas contents, especially the esters contents. Overall, T2 treatment had a better effect than T1 treatment. This study provided an idea for improving water use efficiency and grape quality.

Modulation of innate immune response to viruses including SARS-CoV-2 by progesterone.

Whether and how innate antiviral response is regulated by humoral metabolism remains enigmatic. We show that viral infection induces progesterone via the hypothalamic-pituitary-adrenal axis in mice. Progesterone induces downstream antiviral genes and promotes innate antiviral response in cells and mice, whereas knockout of the progesterone receptor PGR has opposite effects. Mechanistically, stimulation of PGR by progesterone activates the tyrosine kinase SRC, which phosphorylates the transcriptional factor IRF3 at Y107, leading to its activation and induction of antiviral genes. SARS-CoV-2-infected patients have increased progesterone levels, and which are co-related with decreased severity of COVID-19. Our findings reveal how progesterone modulates host innate antiviral response, and point to progesterone as a potential immunomodulatory reagent for infectious and inflammatory diseases.

Physical activity among senior primary school students on weekends in Beijing / 中国学校卫生

Objective@#To analyze the duration and influencing factors of moderate and vigorous physical activity(MVPA) on weekends for primary school students in grades 4 to 6 in Beijing, and to provide a reference for formulating health education and promotion measures.@*Methods@#Multi stage stratified cluster sampling method was used to randomly select 2 515 students from grades 4-6 in 14 primary schools in Beijing, and a self administered questionnaire was used to record MVPA on weekend, social demographic characteristics, other related health behaviors and knowledge. Multivariate Logistic regression analysis was used to explore the influencing factors of MVPA on weekends.@*Results@#The prevalence of insufficient MVPA on weekends in Beijing was 63.54 %, and the prevalence was higher among girls (69.92%) than boys (57.81%) ( χ 2=39.65, P <0.01). Multiple Logistic regression analysis revealed that girls ( OR =1.74), living in rural areas ( OR =1.41), participants attending general schools ( OR = 1.34 ), from divorced family ( OR =1.46), and short sleep duration ( OR =1.50) were more likely to fail to meet the MVPA recommendations( P <0.05).@*Conclusion@#It is quite common that no sufficient weekend MVPA among senior primary school students, among them, the outer suburbs and schools with relatively weak teaching resources are the key places that need attention, and girls are the key groups that need attention.

[Roles of ERK/JNK in carbon black induced AP-1 cell signaling pathway changes].

OBJECTIVE: To investigate the role of ERK/JNK in the alteration of activator protein-1(AP-1) signaling pathway in human embryonic lung fibroblasts(HELFs) induced by carbon black. METHODS: HELFs were cultured in RPMI 1640 medium containing 0, 15, 30, 60, 120 or 240 µg/mL carbon black for 24 h, and the appropriate dose of carbon black was determined by MTT assay result. HELFs were divided into three groups: HELFs, HELFs transfected with ERK dominant negative mutant plasmid(DN-ERK) and HELFs transfected with JNK dominant negative mutant plasmid(DN-JNK). 100 µg/mL carbon black was used to treat HELFs(CB), DN-ERK HELFs(CB-DN-ERK), DN-JNK HELFs(CB-DN-JNK), and HELFs without any treatment were considered as control group. At 0, 1, 2, 4, 8, 12, 24 and 36 h of CB and control groups HELFs, the western blot was used to detect ERK, p-ERK, JNK, p-JNK, p38, p-p38, c-Jun, p-c-Jun, c-Fos, p-c-Fos protein expression levels, and AP-1 activity was detected by luciferase method. Whereas CB-DN-ERK and CB-DN-JNK HELFs were detected only at 24 h. RESULTS: Compared with the protein expression levels at 0 h, CB group HELFs ERK and p-ERK protein expression increased at each time point, whereas p38 protein expression decreased. AP-1 activity of CB group HELFs was declined to the lowest at 8 h(0.72±0.12), and upregulated to the peak at 36 h(1.38±0.11). CB group HELFs c-Fos, p-c-Fos and c-Jun protein expression levels at each time point from 1 h to 24 h were greater than those of 0 h, and p-c-Jun protein expression levels at 1 h, 2 h, 4 h, 8 h, 36 h were also greater than those of 0 h. CB group HELFs AP-1 activity, ERK, p-ERK, JNK, p-JNK, p38, p-p38, c-Jun, p-c-Jun, c-Fos, p-c-Fos protein expression levels changes followed biphasic patterns. There were no statistically significant differences in AP-1 activity between CB group HELFs(1.03±0.10) and CB-DN-ERK group(1.02±0.04) or CB-DN-JNK group(1.09±0.10) HELFs(t=0.16, P=0.88; t=0.73, P=0.50). However, compared with CB group HELFs, c-Fos(t=5.31, P=0.01), p-c-Fos(t=4.33, P=0.01), p-c-Jun(t=10.95, P& lt; 0.01)in CB-DN-JNK group, and c-Fos protein expression levels in CB-DN-ERK group(t=42.72, P& lt; 0.01)were significantly decreased. CONCLUSION: While carbon black induces HELFs increased protein expression levels of ERK, p-ERK, c-Jun, p-c-Jun, c-Fos and p-c-Fos, JNK may upregulate c-Fos, p-c-Fos, p-c-Jun protein expression levels, and ERK may upregulate c-Fos protein expression level.

Recombinant Protein Stability in Cyanobacteria.

The living cell possesses extraordinary molecular and biochemical mechanisms by which to recognize and efficiently remove foreign, damaged, or denatured proteins. This essential function has been a barrier to the overexpression of recombinant proteins in most expression systems. A notable exception is the overexpression in E. coli of recombinant proteins, most of which, however, end-up as "inclusion bodies", i.e., cytoplasmic aggregates of proteins that are inaccessible to the cell's proteasome. "Fusion constructs as protein overexpression vectors" proved to be unparalleled in their ability to cause substantial accumulation of recombinant proteins from plants, animals, and bacteria, as soluble proteins in unicellular cyanobacteria. Recombinant protein levels in the range of 10-20% of the total cellular protein can be achieved. The present work investigated this unique property in the context of recombinant protein stability in Synechocystis sp. PCC 6803 by developing and applying an in vivo cellular tobacco etch virus cleavage system with the objective of separating the target heterologous proteins from their fusion leader sequences. The work provides new insights about the overexpression, cellular stability, and exploitation of transgenes with commercial interest, highly expressed in a cyanobacterial biofactory. The results support the notion that eukaryotic plant- and animal-origin recombinant proteins are unstable, when free in the cyanobacterial cytosol but stable when in a fusion configuration with a highly expressed cyanobacterial native or heterologous protein. Included in this analysis are recombinant proteins of the plant isoprenoid biosynthetic pathway (isoprene synthase, ß-phellandrene synthase, geranyl diphosphate synthase), the human interferon protein, as well as prokaryotic proteins (tetanus toxin fragment C and the antibiotic resistance genes kanamycin and chloramphenicol). The future success of synthetic biology approaches with cyanobacteria and other systems would require overexpression of pathway enzymes to attain product volume, and the work reported in this paper sets the foundation for such recombinant pathway enzyme overexpression.

[Effect of ERK/JNK cell signaling pathway in carbon black induced cytokine IL-6 and IL-8 expression changes].

OBJECTIVE: To investigate the roles of extracellular signal-regulated kinase(ERK)/c-Jun amino-terminal kinase(JNK) signaling pathway on the expression of interleukin-6(IL-6) and interleukin-8(IL-8) in human embryonic lung fibroblasts(HELF) induced by carbon black. METHODS: HELFs were cultured in RPMI-1640 medium containing 0, 15, 30, 60, 120 or 240 µg/mL carbon black for 24 h, and the appropriate dose of carbon black was determined by MTT assay result HELFs were divided into three groups: HELFs, HELFs transfected with ERK dominant negative mutant plasmid(DN-ERK) and HELFs transfected with JNK dominant negative mutant plasmid(DN-JNK). 100 µg/mL carbon black was used to treat HELFs(CB), DN-ERK HELFs(CB-DN-ERK), DN-JNK HELFs(CB-DN-JNK), and HELFs without any black carbon treatment were considered as control group. At 16 h after carbon black treatment, scanning electron microscope(SEM) was used to observe HELFs morphology and whether there were carbon black particless. At 0, 1, 2, 4, 8, 12, 24 and 36 h, the enzyme linked immunosorbent assay(ELISA) was used to detect CB and control groups HELFs IL-6 and IL-8 expression levels, whereas CB-DN-ERK and CB-DN-JNK HELFs were detected only at 24 h. RESULTS: SEM result showed no carbon black particles were observed in CB group HELFs, whereas their surface projections were increased. The CB group HELFs IL-6 expression levels at 2 h(44. 86±3. 65 ng/L) and 4 h(76. 52±3. 15 ng/L) were significantly lower than those of the control group(96. 78±2. 82 and 147. 32±3. 26 ng/L)(P<0. 05), whereas the IL-6 expression levels were significantly higher than those of the control group(105. 54±6. 10, 101. 27±5. 84 and 97. 15±5. 12 ng/L) at 16 h(202. 64±7. 20 ng/L), 24 h(200. 38±6. 20 ng/L) and 36 h(183. 54±4. 54 ng/L)(P<0. 001). At 24 h(136. 75±3. 81 ng/L) and 36 h(149. 12±2. 74 ng/L), the CB group IL-8 expression levels were significantly higher than those of the control group(75. 16±2. 84 and 73. 44±2. 15 ng/L)(P<0. 001). Compared with CB group HELFs, CB-DN-ERK and CB-DN-JNK groups HELFs had significantly lower IL-6 and IL-8 expression levels(P<0. 05). CONCLUSION: While carbon black induced HELFs IL-6 and IL-8 expression levels changes, ERK and JNK may upregulate IL-6 and IL-8 expression levels.

Single Nucleotide Polymorphism Detection for Peach Gummosis Disease Resistance by Genome-Wide Association Study.

Peach gummosis is one of the most widespread and destructive diseases. It causes growth stunting, yield loss, branch, trunk, and tree death, and is becoming a restrictive factor in healthy and sustainable development of peach production. Although a locus has been identified based on bi-parental quantitative trait locus (QTL) mapping, selection of gummosis-resistant cultivars remains challenging due to the lack of resistant parents and of the complexity of an inducing factor. In this study, an integrated approach of genome-wide association study (GWAS) and comparative transcriptome was used to elucidate the genetic architecture associated with the disease using 195 accessions and 145,456 genome-wide single nucleotide polymorphisms (SNPs). The broad-sense and narrow-sense heritabilities were estimated using 2-year phenotypic data and genotypic data, which gave high values of 70 and 73%, respectively. Evaluation of population structure by neighbor-joining and principal components analysis (PCA) clustered all accessions into three major groups and six subgroups, mainly according to fruit shape, hairy vs. glabrous fruit skin, pedigree, geographic origin, and domestication history. Five SNPs were found to be significantly associated with gummosis disease resistance, of which SNPrs285957, located on chromosome6 across 28 Mb, was detected by both the BLINK and the FarmCPU model. Six candidate genes flanked by or harboring the significant SNPs, previously implicated in biotic stress tolerance, were significantly associated with this resistance. Two highly resistant accessions were identified with low disease severity, which could be potential sources of resistance genes for breeding. Our results provide a fresh insight into the genetic control of peach gummosis disease.