Ruthenium-Catalyzed Carbonylation of α-Aminoaryl-Tethered Alkylidenecyclopropanes: Synthesis of Eight-Membered Benzolactams.

The synthesis of medium-sized lactams is a great challenge because of the unfavorable transannular interactions and entropic barriers in the transition state. We have developed a ruthenium-catalyzed carbonylation of α-aminoaryl-tethered alkylidenecyclopropanes (ACPs) that allows for the efficient preparation of valuable eight-membered benzolactams under ligand-free conditions. The amino group served a dual role of both directing group and nucleophile to facilitate the metallacycle formation and the carbonylation.

An ultra-compact acoustofluidic device based on the narrow-path travelling surface acoustic wave (np-TSAW) for label-free isolation of living circulating tumor cells.

Obtaining highly purified intact living cells from complex environments has been a challenge, such as the isolation of circulating tumor cells (CTCs) from blood. In this work, we demonstrated an acoustic-based ultra-compact device for cell sorting, with a chip size of less than 2 × 1.5 cm2. This single actuator device allows non-invasive and label-free isolation of living cells, offering greater flexibility and applicability. The device performance was optimized with different-sized polystyrene (PS) particles and blood cells spiked with cancer cells. Using the narrow-path travelling surface acoustic wave (np-TSAW), precise isolation of 10 µm particles from a complex mixture of particles (5, 10, 20 µm) and separation of 8 µm and 10 µm particles was achieved. The purified collection of 10 µm particles with high separation efficiency (98.75%) and high purity (98.1%) was achieved by optimizing the input voltage. Further, we investigated the isolation and purification of CTCs (MCF-7, human breast cancer cells) from blood cells with isolation efficiency exceeding 98% and purity reaching 93%. Viabilities of the CTCs harvested from target-outlet were all higher than 97% after culturing for 24, 48, and 72 h, showing good proliferation ability. This novel ultra-miniaturized microfluidic chip demonstrates the ability to sorting cells with high-purity and label-free, providing an attractive miniaturized system alternative to traditional sorting methods.

Rutaecarpine attenuates high glucose-induced damage in AC16 cardiomyocytes by suppressing the MAPK pathway.

Diabetic cardiomyopathy is a common diabetic complication, resulting in heart failure. Rutaecarpine is an active compound with cardiovascular protective effects. However, the function of rutaecarpine in diabetic cardiomyopathy is largely unknown. The aim of this research was to study the effect and action mechanism of rutaecarpine in high glucose (HG)-induced cardiomyocyte damage. The overlapping genes of diabetic cardiomyopathy and rutaecarpine were analyzed according to GeneCards, DisGeNet, and SwissTargetPrediction. Cell damage was investigated by determining apoptosis, oxidative stress, and inflammatory response in HG-stimulated AC16 cells. The expression of proteins involved in the mitogen-activated protein kinase (MAPK) signaling was measured using Western blotting. Totally seven overlapping genes of diabetic cardiomyopathy and rutaecarpine were screened out and predicted to be associated with the MAPK signaling. Rutaecarpine protected against HG-induced cardiomyocyte damage by enhancing cell viability and reducing cell apoptosis, caspase-3 activity, and lactate dehydrogenase (LDH) release. Rutaecarpine mitigated HG-induced oxidative stress in cardiomyocytes through decreasing reactive oxygen species (ROS) formation and malondialdehyde (MDA) level and elevating superoxide dismutase (SOD) activity and glutathione peroxidase (GSH-Px) level. Rutaecarpine alleviated HG-induced inflammatory response via reducing the level of interleukin (IL)-1ß, IL-6, tumor necrosis factor (TNF)-α, and IL-8. Moreover, rutaecarpine inhibited HG-induced activation of the MAPK pathway. Treatment with MAPK signaling agonist reversed the suppressive effect of rutaecarpine on HG-induced damage. In conclusion, rutaecarpine alleviated HG-induced cardiomyocyte damage through decreasing apoptosis, oxidative stress, and inflammatory response by inactivating the MAPK pathway.

A Novel Detachable, Reusable, and Versatile Acoustic Tweezer Manipulation Platform for Biochemical Analysis and Detection Systems.

Multifunctional, integrated, and reusable operating platforms are highly sought after in biochemical analysis and detection systems. In this study, we demonstrated a novel detachable, reusable acoustic tweezer manipulation platform that is flexible and versatile. The free interchangeability of different detachable microchannel devices on the acoustic tweezer platform was achieved by adding a waveguide layer (glass) and a coupling layer (polydimethylsiloxane (PDMS) polymer film). We designed and demonstrated the detachable multifunctional acoustic tweezer platform with three cell manipulation capabilities. In Demo I, the detachable acoustic tweezer platform is demonstrated to have the capability for parallel processing and enrichment of the sample. In Demo II, the detachable acoustic tweezer platform with capability for precise cell alignment is demonstrated. In Demo III, it was demonstrated that the detachable acoustic tweezer platform has the capability for the separation and purification of cells. Through experiments, our acoustic tweezer platform has good acoustic retention ability, reusability, and stability. More capabilities can be expanded in the future. It provides a simple, economical, and multifunctional reusable operating platform solution for biochemical analysis and detection systems.

Brain Morphological and Functional Changes in Adenomyosis with Pain: A Resting State Functional Magnetic Resonance Imaging Study.

The absence of clinically objective methods to evaluate adenomyosis-associated pain and the poor understanding of its pathophysiology lead to treatment limitations. We conducted a resting-state functional magnetic resonance imaging study with 49 patients with pain-related adenomyosis and 30 pain-free controls to investigate brain morphological alterations and regional dysfunctions in patients with pain-related adenomyosis. These patients had significantly higher scores for anxiety and depression than the control group (p < 0.05). They also had a lower gray matter volume (GMV) in the bilateral insula, left angular gyrus, precuneus, left inferior temporal gyrus, and left postcentral gyrus (p < 0.05, AlphaSim corrected). Similarly, decreased voxel-mirrored homotopic connectivity was observed in the bilateral insula, posterior cingulate cortex, middle frontal gyrus, and postcentral gyrus in the adenomyosis patient group (p < 0.05, AlphaSim corrected). Regional homogeneity showed significant differences mainly in the bilateral cerebellum, left inferior frontal gyrus, medial prefrontal cortex, and posterior cingulate gyrus. Correlation analysis showed that the degree of depression in patients with adenomyosis was negatively correlated with the GMV of the left angular gyrus. The results show that these patients exhibited changes in multiple brain regions associated with pain as well as emotion perception and processing.

Effect and molecular mechanism research of Astragalus membranaceus on inhibiting intestinal absorption of six alkaloids of Aconitum carmichaelii in spleen deficiency rats.

Objective: To investigate the effect and the mechanism of Astragalus membranaceus (Huangqi in Chinese, HQ) extract on the intestinal absorption of six alkaloids of Aconitum carmichaelii (Fuzi in Chinese, FZ) in rats with spleen deficiency and provide novel insights into the application of HQ on modulating intestinal barrier. Methods: Four-week-old male Sprague-Dawley rats were fed with Xiaochengqi Decoction to induce the spleen deficiency model for 40 d. Single-pass intestinal perfusion model were used to study the effects of HQ extract on the absorption of alkaloids. Protein expression and mRNA levels of MRP2 and BCRP and tight junction proteins (TJ, including Claudin-1, Occludin and ZO-1) were measured using Western blot and real-time PCR, respectively. The location and expression of TJ protein was also investigated by the immunofluorescence method. Results: Compared with the normal group, the protein expression of MRP2, BCRP and TJ proteins in the model group were significantly down-regulated. After oral administration of HQ, the alkaloid absorption in intestinal villi was inhibited, MRP2, BCRP and TJ proteins were up-regulated, the green fluorescence staining of Claudin-1, Occludin, and ZO-1 was enhanced, and a thick layer of mucus was deposited on the surface of the epithelium of the intestinal cavity. Conclusion: HQ as an intestinal barrier modulator improves the physiological changes of the intestinal environment of spleen deficiency to reduce the absorption of toxic components, leading to a decrease in the absorption of drug-like molecules.

A Perturbed Asymmetrical Y-TypeSheathless Chip for Particle Control Based on Adjustable Tilted-Angle Traveling Surface Acoustic Waves (ataTSAWs).

The precise control of target particles (20 µm) at different inclination angles θi is achieved by combining a perturbed asymmetric sheathless Y-type microchannel and a digital transducer (IDT). The offset single-row micropillar array with the buffer area can not only concentrate large and small particles in a fixed region of the flow channel, but also avoid the large deflection of some small particles at the end of the array. The addition of the buffer area can effectively improve the separation purity of the chip. By exploring the manufacturing process of the microchannel substrate, an adjustable tilted-angle scheme is proposed. The use of ataTSAW makes the acoustic field area in the microchannel have no corner effect region. Through experiments, when the signal source frequency was 33.6 MHz, and the flow rate was 20 µL/min, our designed chip could capture 20 µm particles when θi = 5°. The deflection of 20 µm particles can be realized when θi = 15°-45°. The precise dynamic separation of 20 µm particles can be achieved when θi = 25°-45°, and the separation purity and efficiency were 97% and 100%, respectively.

Enhanced Detection in Droplet Microfluidics by Acoustic Vortex Modulation of Particle Rings and Particle Clusters via Asymmetric Propagation of Surface Acoustic Waves.

As a basis for biometric and chemical analysis, issues of how to dilute or concentrate substances such as particles or cells to specific concentrations have long been of interest to researchers. In this study, travelling surface acoustic wave (TSAW)-based devices with three frequencies (99.1, 48.8, 20.4 MHz) have been used to capture the suspended Polystyrene (PS) microspheres of various sizes (5, 20, 40 µm) in sessile droplets, which are controlled by acoustic field-induced fluid vortex (acoustic vortex) and aggregate into clusters or rings with particles. These phenomena can be explained by the interaction of three forces, which are drag force caused by ASF, ARF caused by Leaky-SAW and varying centrifugal force. Eventually, a novel approach of free transition between the particle ring and cluster was approached via modulating the acoustic amplitude of TSAW. By this method, multilayer particles agglomerate with 20 µm wrapped around 40 µm and 20 µm wrapped around 5 µm can be obtained, which provides the possibility to dilute or concentrate the particles to a specific concentration.

Higher Risk of Anxiety and Depression in Women with Adenomyosis as Compared with Those with Uterine Leiomyoma.

The main symptoms of adenomyosis may adversely affect physical and mental health and quality of life (QOL). However, studies are few on this topic. This study evaluated the effect of adenomyosis on anxiety, depression, and QOL. Participants with adenomyosis (n = 90) or leiomyoma (n = 59) completed questionnaires, including the visual analog scale (VAS) for pain, Hospital Anxiety and Depression Scale (HADS), and Short Form (SF)-36. HADS anxiety and depression positive rates, physical (PCS) and mental (MCS) component summary scores, and eight subscale scores of SF-36 were compared between the two groups. Among patients with adenomyosis, the incidence of anxiety symptoms was 28.9% (control group, 10.2%; OR = 3.589, 95% CI: 1.375-9.367), with 10% of patients showing moderate-to-severe symptoms. The incidence of depressive symptoms was 14.4% (control group, 3.4%; OR = 4.812, 95% CI: 1.044-22.168). The case group had significantly lower PCS and MCS scores than the control group. In patients with adenomyosis, being employed (OR = 6.393, 95% CI: 1.153-35.440) and perianal pain (OR = 25.419, 95% CI: 2.504-258.024) were risk factors for anxiety, and perianal pain (OR = 15.208, 95% CI: 3.050-75.836) was a risk factor for depression. Compared with leiomyoma, adenomyosis is associated with a higher risk of anxiety and depression, with a poorer QOL.

A Novel Perturbed Spiral Sheathless Chip for Particle Separation Based on Traveling Surface Acoustic Waves (TSAW).

The combination of the new perturbed spiral channel and a slanted gold interfingered transducer (IDT) is designed to achieve precise dynamic separation of target particles (20 µm). The offset micropillar array solves the defect that the high-width flow (avoiding the occurrence of channel blockage) channel cannot realize the focusing of small particles (5 µm, 10 µm). The relationship between the maximum design gap of the micropillar (Smax) and the particle radius (a) is given: Smax = 4a, which not only ensures that small particles will not pass through the micropillar gap, but also is compatible with the appropriate flow rates. A non-offset micropillar array was used to remove 20 µm particles in the corner area. The innovation of a spiral channel structure greatly improves the separation efficiency and purity of the separation chip. The separation chip designed by us achieves deflection separation of 20 µm particles at 24.95-41.58 MHz (κ = 1.09-1.81), at a flow rate of 1.2 mL per hour. When f = 33.7 MHz (κ = 1.47), the transverse migration distance of 20 µm particles is the smallest, and the separation purity and efficiency are as high as 92% and 100%, respectively.

Macrophage-associated immune checkpoint CD47 blocking ameliorates endometriosis.

Peritoneal macrophages play a significant role in the progression of endometriosis (EM), but their functional differentiation is still unclear, and their phagocytic ability is weak. CD47-signal-regulated protein α (SIRPα) and PD-L1-PD-1 are considered immune checkpoints associated with macrophage phagocytosis. A specific blockade of these two pathways had been shown to increase the phagocytic clearance of cancer cells by macrophages in most cancers. We hypothesized that targeting CD47/PD-L1 in EM could improve the phagocytosis of macrophages, thereby delaying the progression of EM. From localization to quantification, from mRNA to protein, we comprehensively evaluated the expression of CD47 and PD-L1 in EM. We demonstrated that the CD47 expression in ectopic endometrium from patients with EM was significantly increased, but PD-L1 was not. We performed direct co-culture experiments of endometrial stromal cells with macrophages in vitro and in vivo to assess whether ectopic endometrial stromal cells escape macrophage phagocytosis through the CD47-SIRPα signaling pathway. The results showed that targeting CD47 increased the phagocytic capacity of macrophages. Interestingly, we also found that the reduction of CD47 expression promoted apoptosis of endometrial stromal cells. In conclusion, these data suggested that targeting CD47 can effectively target ectopic endometrial stromal cells through a dual mechanism of increased phagocytosis of macrophages and induced apoptosis of ectopic endometrial stromal cells. Thus, immunotherapy based on the CD47-SIRPα signaling pathway has some potential in treating EM, but further mechanistic studies are needed to explore more effective and specific antibodies.

Chemosensitizing effect and mechanism of imperatorin on the anti-tumor activity of doxorubicin in tumor cells and transplantation tumor model.

Multidrug resistance of tumors has been a severe obstacle to the success of cancer chemotherapy. The study wants to investigate the reversal effects of imperatorin (IMP) on doxorubicin (DOX) resistance in K562/DOX leukemia cells, A2780/Taxol cells and in NOD/SCID mice, to explore the possible molecular mechanisms. K562/DOX and A2780/Taxol cells were treated with various concentrations of DOX and Taol with or without different concentrations of IMP, respectively. K562/DOX xenograft model was used to assess anti-tumor effect of IMP combined with DOX. MTT assay, Rhodamine 123 efflux assay, RT-PCR, and Western blot analysis were determined in vivo and in vitro. Results showed that IMP significantly enhanced the cytotoxicity of DOX and Taxol toward corresponding resistance cells. In vivo results illustrated both the tumor volume and tumor weight were significantly decreased after 2-week treatment with IMP combined with DOX compared to the DOX alone group. Western blotting and RT-PCR analyses indicated that IMP downregulated the expression of P-gp in K562/DOX xenograft tumors in NOD/SCID mice. We also evaluated glycolysis and glutamine metabolism in K562/DOX cells by measuring glucose consumption and lactate production. The results revealed that IMP could significantly reduce the glucose consumption and lactate production of K562/DOX cells. Furthermore, IMP could also remarkably repress the glutamine consumption, α-KG and ATP production of K562/DOX cells. Thus, IMP may sensitize K562/DOX cells to DOX and enhance the anti-tumor effect of DOX in K562/DOX xenograft tumors in NOD/SCID mice. IMP may be an adjuvant therapy to mitigate the multidrug resistance in leukemia chemotherapy.

Continuous Particle Separation Driven by 3D Ag-PDMS Electrodes with Dielectric Electrophoretic Force Coupled with Inertia Force.

Cell separation has become @important in biological and medical applications. Dielectrophoresis (DEP) is widely used due to the advantages it offers, such as the lack of a requirement for biological markers and the fact that it involves no damage to cells or particles. This study aimed to report a novel approach combining 3D sidewall electrodes and contraction/expansion (CEA) structures to separate three kinds of particles with different sizes or dielectric properties continuously. The separation was achieved through the interaction between electrophoretic forces and inertia forces. The CEA channel was capable of sorting particles with different sizes due to inertial forces, and also enhanced the nonuniformity of the electric field. The 3D electrodes generated a non-uniform electric field at the same height as the channels, which increased the action range of the DEP force. Finite element simulations using the commercial software, COMSOL Multiphysics 5.4, were performed to determine the flow field distributions, electric field distributions, and particle trajectories. The separation experiments were assessed by separating 4 µm polystyrene (PS) particles from 20 µm PS particles at different flow rates by experiencing positive and negative DEP. Subsequently, the sorting performances of the 4 µm PS particles, 20 µm PS particles, and 4 µm silica particles with different solution conductivities were observed. Both the numerical simulations and the practical particle separation displayed high separating efficiency (separation of 4 µm PS particles, 94.2%; separation of 20 µm PS particles, 92.1%; separation of 4 µm Silica particles, 95.3%). The proposed approach is expected to open a new approach to cell sorting and separating.

A cohort study of the efficacy of the dienogest and the gonadotropin-releasing hormone agonist in women with adenomyosis and dysmenorrhea.

PURPOSE: To study the efficacy and safety of the dienogest and the gonadotropin-releasing hormone agonist (GnRH-a) in symptomatic females with uterine adenomyosis. METHODS: A total of 127 patients with adenomyosis with a chief complaint of dysmenorrhea were recruited. The first group received 2 mg of dienogest (DNG) daily, whereas the second group received goserelin acetate (GS) (3.6 mg/4 weeks) for 12 weeks. Outpatient follow-up was undertaken after 12 weeks. RESULTS: Among 127 women, 56/63 (88.9%) patients completed the treatment in the DNG group, whereas 62/64 (96.9%) patients completed the treatment in the GS group. A significant decrease in dysmenorrhea symptoms as measured by the visual analog scale (VAS) and Carcinoma antigen125 (CA125) after 12 weeks of treatment was observed in both groups (p < .001). The hemoglobin of anemic patients did not significantly improve after 12 weeks of treatment (p＝0.21) and the uterine volume slightly increased without statistical significance (p＝0.10) in the DNG group. Simultaneously, The hemoglobin of anemic patients significantly improved (p < .001) and the uterine volume significantly decreased (p < .001) in the GS group. CONCLUSIONS: Dienogest effectively alleviates the symptoms of dysmenorrhea in patients with adenomyosis, but it cannot improve the anemia or reduce the size of the uterus. GnRH-a is more effective in improving anemia and reducing the uterine volume in patients with adenomyosis. TRIAL REGISTRATION: ChiCTR1900024958.

Magnetic graphene oxide-containing chitosan­sodium alginate hydrogel beads for highly efficient and sustainable removal of cationic dyes.

Developing recyclable and efficient adsorbents for cationic dyes removal from wastewater is crucial for ensuring green ecology and drinking water safety. Herein, we demonstrated a novel magnetic gel bead adsorbent that was synthesized by employing graphene oxide (GO) modified Fe3O4 as magnetic nanoparticles doped sodium alginate (SA)/chitosan (CS) gel (SA/GO@Fe3O4/CS). The GO@Fe3O4 sample was prepared based on GO by the chemical co-precipitation method, which not only reduced the aggregation of Fe3O4 but also increases the specific surface area of the composite gel beads. The prepared gel beads were used to adsorb methylene blue (MB), neutral red (NR), and safranine T (ST). The experimental results showed that the adsorption capacity of SA/GO@Fe3O4/CS gel beads for MB, NR, and ST reached 21.325 mg/g, 44.654 mg/g and 44.313 mg/g. After five recycles, the removal rates could still reach more than 90% of the original, exhibiting a high recovery rate. Therefore, this paper provides a strategy for the preparation of high efficiency and recyclable cationic dye adsorbents with a large specific surface area.

Metabolite profiles in the peritoneal cavity of endometriosis patients and mouse models.

RESEARCH QUESTION: Which metabolites are altered in the peritoneal cavity of women with endometriosis? Could the mouse endometriosis model simulate these alterations? DESIGN: Thirteen women with endometriosis and seven women with other benign gynaecological diseases, who underwent laparoscopic surgery, were included in this study. None had received hormonal therapy for 3 months before surgery. For the animal experiments, six and five mice were included in the endometriosis and control groups, respectively. Peritoneal fluid from the patients and peritoneal lavage fluid from the mice was collected and analysed. Non-targeted metabolomics via liquid chromatography with tandem mass spectrometry was used to identify the altered metabolites in the peritoneal fluid of endometriosis patients and mouse models. MetaboAnalyst 4.0 was used to visualize the data. RESULTS: Several metabolites in the peritoneal cavity were significantly altered in both humans and mice with endometriosis. Concentrations of lysophosphatidylcholine (LysopC) (P=0.017 in patients and P=0.041 in the mouse model) and derivatives of phosphoethanolamine (1-arachidonoyl-sn-glycero-3-phosphoethanolamine in patients, P=0.027; 1-oleoyl-sn-glycero-3-phosphoethanolamine in patients, P=0.0086; and phosphorylethanolamine in the mouse model, P=0.0027) were significantly up-regulated in both, whereas concentrations of acylcarnitines (l-palmitoylcarnitine, P=0.047; and stearoylcarnitine, P=0.029) and kynurenine (P=0.045) were significantly increased only in humans. The human and mouse samples shared three altered enriched metabolite sets. CONCLUSIONS: Women with endometriosis show an altered metabolic state in the abdominal cavity. The endometriosis mouse model shared half of the significantly altered metabolite sets found in the abdominal cavity of humans.

M1 Macrophage-Derived Nanovesicles Repolarize M2 Macrophages for Inhibiting the Development of Endometriosis.

Background: Endometriosis is a common nonmalignant gynecological disorder that affects 10-15% women of reproductive age and causes several symptoms that result in decreased quality of life and a huge social burden. In recent decades, extracellular vesicles (EVs) have gained attention as a potential therapeutic tool; however, the therapeutic effects of EVs against endometriosis have not been reported. Accordingly, in this study, we investigated the feasibility of nanovesicles (NVs) derived from M1 macrophages (M1NVs) in treating endometriosis. Methods: M1NVs were prepared by serial extrusion. Co-culture assays were performed to investigate changes in tube formation and migration/invasion of eutopic endometrial stroma cells (ESCs) obtained from patients with endometriosis (EM-ESCs). A mouse model of endometriosis was established, and mice were treated with phosphate-buffered saline, M0NVs, or M1NVs to evaluate the efficacy and safety of M1NV for treating endometriosis. Results: M1NVs directly or indirectly inhibited the migration and invasion of EM-ESCs and reduced tube formation. In the mouse model, M1NVs suppressed the development of endometriosis through reprogramming of M2 macrophages, without causing damage to the organs. Conclusions: M1NVs inhibit the development of endometriosis directly, or through repolarizing macrophages from M2 to M1 phenotype. Hence, administration of M1NVs may represent a novel method for the treatment of endometriosis.

Overexpression of the Apple (Malus × domestica) MdERF100 in Arabidopsis Increases Resistance to Powdery Mildew.

APETALA2/ETHYLENE RESPONSIVE FACTOR (AP2/ERF) transcription factors play important roles in plant development and stress response. Although AP2/ERF genes have been extensively investigated in model plants such as Arabidopsis thaliana, little is known about their role in biotic stress response in perennial fruit tree crops such as apple (Malus × domestica). Here, we investigated the role of MdERF100 in powdery mildew resistance in apple. MdERF100 localized to the nucleus but showed no transcriptional activation activity. The heterologous expression of MdERF100 in Arabidopsis not only enhanced powdery mildew resistance but also increased reactive oxygen species (ROS) accumulation and cell death. Furthermore, MdERF100-overexpressing Arabidopsis plants exhibited differential expressions of genes involved in jasmonic acid (JA) and salicylic acid (SA) signaling when infected with the powdery mildew pathogen. Additionally, yeast two-hybrid and bimolecular fluorescence complementation assays confirmed that MdERF100 physically interacts with the basic helix-loop-helix (bHLH) protein MdbHLH92. These results suggest that MdERF100 mediates powdery mildew resistance by regulating the JA and SA signaling pathways, and MdbHLH92 is involved in plant defense against powdery mildew. Overall, this study enhances our understanding of the role of MdERF genes in disease resistance, and provides novel insights into the molecular mechanisms of powdery mildew resistance in apple.

NAC domain gene VvNAC26 interacts with VvMADS9 and influences seed and fruit development.

Grapevine (Vitis vinifera), as one of the potential gardening fruit have important economic and nutritional values. Seedless grapes are popular due to their convenience and excellent taste. As multifunctional proteins, the NAC family involves hormonal pathways, plant aging, and biological stress. However, reports of NAC affecting seed development are rare. Here, the role of grapevine VvNAC26 in regulating fruit ripening and seed size was characterized. There were remarkable differences in the expression of VvNAC26 in seeded and seedless grape varieties during ovule development. The exogenous transformation of VvNAC26 in tomato decreased the cells size of pericarp, fruits and seeds. In addition, led to cotyledon cells arranged more closely and narrowly and obviously decreased seeds at the fruit ripening stage. The tomato fruit of transgenic lines was darker red and underwent color conversion earlier than that of the wild type in the same period. Furthermore, the expression of some genes associated with hormone and fruit development was changed in overexpressed lines. Yeast two-hybrid and BiFC assays showed that VvNAC26 interacted with VvMADS9. In conclusion, these results suggest that VvNAC26 may regulate fruit and seed development by influencing multiple hormone pathways and interacting with VvMADS9 in grape. VvNAC26 may also serve as a candidate for future understanding of the potential regulatory mechanism of seed development and molecular breeding in grapevine.

Comparative Study of Volatile Compounds and Expression of Related Genes in Fruit from Two Apple Cultivars during Different Developmental Stages.

Aromatic volatile compounds are important contributors to fruit quality that vary among different cultivars. Herein, headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry was used to determine changes in volatile compounds and related gene expression patterns in "Ruixue" and "Fuji" apples (Malus domestica Borkh.) during fruit development and maturation. Volatile compounds detected in the fruit of both cultivars exhibited similar trends across different developmental stages. In the early stages of "Ruixue" fruit development (60 days after full bloom), there were fewer volatile compounds, mainly aldehydes (87.0%). During fruit maturation (180 days after full bloom), the types and amounts of volatile compounds increased, mainly including esters (37.6%), and alkenes (23.2%). The total volatile concentration, the types of major volatile compounds, and their relative content in both cultivars varied across different stages. Gene expression analysis indicated that the upregulation of MdLOX, MdAAT2, and MdADH3 was associated with increased aroma compound content, especially esters, during fruit development in both cultivars. Changes in the expression of MdArAT, MdACPD, MdADH3, MdAAT2, and MdLOX may lead to differences in volatile compounds between apple cultivars.