BackgroundTrimethylation of histone H3 lysine 27 (H3K27me3) confers a repressive chromatin state and is dynamically deposited and removed to regulate gene expression throughout plant growth and development. Nevertheless, the three-dimensional epigenome architecture linked to H3K27me3 and its regulatory role in controlling rice development remain unclear.ResultsHere, we employ long-read ChIA-PET to map the genome topology associated with H3K27me3 in rice, identifying hundreds of repressive chromatin spatial clusters within chromosomes. The T-DNA insertional mutagenesis of EMBRYONIC FLOWER 2b (OsEMF2b) leads to disruption of H3K27me3-associated chromatin interaction networks. Notably, we discover that the three key flowering time loci, Early heading date 1 (Ehd1) and Heading date 3a (Hd3a)/Rice Flowering Locus T1 (RFT1) form an inter-chromosomal repressive chromatin spatial cluster as a cis-interaction hub. Further investigation reveals that Ghd7, Ghd8, and Hd1 undergo liquid-liquid phase separation and bind to Ehd1-Hd3a/RFT1 repressive chromatin spatial cluster as trans flowering regulators.ConclusionOur findings elucidate a multi-dimensional topological framework of H3K27me3-associated loci in rice and identify a spatial flowering gene cluster regulated by the phase-separated repressor complex of Ghd7, Ghd8, and Hd1. These results expand our knowledge of the structural basis of the three-dimensional genome governing the transcriptional coordination of florigens in rice.

The Ocimum genus, valued for its culinary, medicinal, and aromatic uses, is often cultivated in regions affected by drought and salinity. This study investigates the adaptive responses of QingFeng (QF), a newly identified basil cultivar locally known as vegetable "jingjie" in China, to high salinity and drought stress. DNA barcoding, mucilaginous seed analysis, flow cytometry, and karyotyping confirmed QF's identity, revealing a chromosome number of 2n = 76 and a 1C genome size of 2962 ± 36 Mb. Phenotyping, transcriptomic and metabolomic profiling, and hormone monitoring showed drought primarily suppressed leaf growth, while salinity inhibited root elongation. Notably, convergent and divergent responses to salinity and drought were observed, including stress-specific remodeling of core metabolic pathways such as the GS-GOGAT cycle, photorespiration, TCA cycle, and pyrimidine metabolism. Secondary metabolite profiling further demonstrated stress-specific patterns in the accumulation of flavonoids and alkaloids. These findings provide a theoretical foundation for deciphering abiotic stress adaptation mechanisms in basil and offering practical approaches to enhance local cultivation of the QingFeng cultivar.

This study aimed to evaluate the therapeutic effects of Omentin-1 on osteoarthritis and explore its protective mechanisms. Two osteoarthritis mouse models were created: wild-type and SIRT6−/−. After five weeks of Omentin-1 injection, we evaluated M1 and M2 macrophage distribution in the mouse synovium and measured inflammatory factor levels. Matrix proteins related to cartilage repair were detected, and safranin O-green and toluidine blue staining were used to evaluate the repair. Furthermore, we evaluated how Omentin-1 regulated macrophage polarization and explored potential mechanisms. Omentin-1 was low in osteoarthritis mice and linked to M1 macrophage polarization. It protected these mice by reducing synovial inflammation, decreasing M1 macrophages, increasing M2 macrophages, and lowering pro-inflammatory factors while raising anti-inflammatory factors and minimizing inflammatory cell infiltration in the synovium. Omentin-1 enhanced Collagen II and α-SMA expression in cartilage, aiding repair. Further mechanistic studies indicated that Omentin-1 can enhance the expression of SIRT6 in the joint tissues of mice with osteoarthritis. Omentin-1’s inhibition of inflammation in osteoarthritis mice was linked to SIRT6. Elevated Omentin-1 levels can mitigate the inflammatory response in osteoarthritis by enhancing SIRT6 expression, inhibiting inflammatory factors, suppressing M1 macrophages, and increasing M2 macrophages.

Knee osteoarthritis (KOA) is a degenerative joint disorder characterized by articular cartilage degeneration and synovial inflammation. Dysfunction mediated by autophagy-related protein 5 (ATG5) represents a key driver of KOA pathogenesis, while the cGAS-STING-NLRP3 signaling axis is closely associated with inflammation and apoptosis. Therefore, this study aims to explore the regulatory effect of ATG5 on the cGAS-STING-NLRP3 axis and its specific role in KOA. HE staining, Safranin O-fast green staining were used to assess the degree of cartilage degeneration in KOA rats. TUNEL staining were applied to observe the apoptosis of chondrocytes in cartilage tissues. Levels of inflammatory factors in serum of rats and cells were detected by ELISA. Expression of proteins levels was analyzed using western blot and immunofluorescence assay. Flow cytometry was used to investigate the apoptosis. In cartilage tissues of KOA rats, ATG5 was lowly expressed, while cGAS, STING and NLRP3 were overexpressed. Co-localization was observed between ATG5 and STING. Overexpression of ATG5 led to decreased expression of inflammatory factors, cGAS, STING, NLRP3, p62 and Cleaved caspase-1, a reduced Mankin score, and increased Beclin1 expression. However, knockout of ATG5 reversed these changes. Additionally, overexpression of ATG5 decreased the apoptosis rate of chondrocytes, whereas inhibition of ATG5 promoted chondrocyte apoptosis. In conclusion, ATG5 regulates the cGAS-STING-NLRP3 axis, thereby promoting chondrocyte autophagy and inhibiting inflammation, which in turn protects chondrocytes and alleviates KOA. In conclusion, ATG5 modulates the cGAS-STING-NLRP3 axis, thereby enhancing chondrocyte autophagy and suppressing inflammation, which collectively protects chondrocytes and alleviates the progression of KOA.

Background: This study employed a severed finger rat model to analyze the effects of human mesenchymal stem cells (MSCs) on angiogenesis, inflammatory response, apoptosis, and oxidative stress, to evaluate the possible mechanism of the repair effect of MSCs on severed finger (SF) rats. Methods: Sixty Sprague-Dawley (SD) rats were categorized into five groups (n = 12). The pathological changes of severed finger tissues were investigated by Hematoxylin and eosin (H&E) staining on day 14 after the rats were sacrificed. The levels of inflammatory factors and oxidative stress factors were detected by ELISA. Terminal Deoxynucleotidyl Transferase (TdT) dUTP Nick End Labeling (TUNEL) was employed to assess the apoptosis of chondrocytes in severed finger tissues. The expression of osteocalcin (OCN), osteopontin (OPN), Collagen I (Col-1), and CD31 were detected by immunohistochemistry or immunofluorescence assay, respectively. The expression levels of related proteins were determined by western blot. Result: Our study presented evidence that MSCs treatment improved pathological changes of skin and bone tissue, diminished the inflammatory response, prevented oxidative stress injury, suppressed chondrocyte apoptosis, and promoted angiogenesis, and bone formation compared to the model group. In addition, EX527 treatment attenuated the effect of MSCs, SRT1720 and ML385 co-treatment also attenuated the effect of MSCs. Importantly, the MSCs treatment increased the expression of Sirtuin 1(SIRT1)/Nuclear factor erythroid2-related factor 2(Nrf2) relate proteins. Conclusion: Our study indicated that the mechanism of the effect of MSCs on a severed finger was related to the SIRT1/ Nrf2 signaling pathway.

Purpose:Percutaneous pedicle screw fixation is a common minimally invasive treatment for traumatic thoracolumbar and lumbar fractures; however, research on hardware removal after successful healing is limited. We aimed to introduce a rapid, safe, minimally invasive, and cost-effective method for percutaneous pedicle screw removal.Patients and Methods:We conducted a retrospective analysis of demographic (age, sex, body mass index, alcohol use, and current smoking), clinical (hypertension and diabetes mellitus), surgical (affected levels, number of screws, time of surgery, and blood loss), and treatment cost characteristics of 92 patients who had undergone percutaneous pedicle screw removal between May 2016 and February 2023. The first 57 patients underwent the conventional method, and the remaining 35 underwent the modified method. Independent-sample t-tests and chi-square tests were used to compare continuous and categorical variables, respectively, between the two groups.Results:No significant differences were observed in the demographic parameters, complications, or affected levels between the groups. However, the average surgical time (P=0.000) was significantly shorter, and the average blood loss volume (P=0.002) and total cost (P=0.000) were significantly lower in the modified group than in the conventional group.Conclusion:Compared with the conventional method, our modified method can shorten the surgical time, reduce blood loss, and reduce the total cost of treatment. It is a quick and safe minimally invasive method that does not require additional surgical instruments and is suitable for implementation in primary hospitals.

Osteoarthritis (OA) is a common joint disease that is characterized by inflammation and cartilage degradation. Death-associated protein kinase 1 (DAPK1) is a multi-domain serine/threonine kinase and has been reported to be involved in the progression of OA. However, its role and mechanism in OA remain unclear. Here, we found the expression of DAPK1 in OA cartilage tissues was higher than that in normal cartilage tissues. The expression of DAPK1 in chondrocytes was up-regulated by IL-1β. Knockdown of DAPK1 promoted cell viability and anti-apoptotic protein expression, while it inhibited the apoptosis rate and pro-apoptotic protein expressions in IL-1β-induced chondrocytes. In addition, DAPK1 inhibition reduced the levels of inflammatory cytokines and expressions of matrix metalloproteinases (MMPs), and increased the expressions of collagen II and aggrecan. The data of mechanistic investigation indicated that the expression of pigment epithelium-derived factor (PEDF) was positively regulated by DAPK1. Overexpression of PEDF attenuated the effects of DAPK1 knockdown on IL-1β-induced cell viability, apoptosis, inflammation, and cartilage degradation. Furthermore, PEDF overexpression restored the activity of the NF-κB pathway and NLRP3 inflammasome after DAPK1 knockdown. Collectively, down-regulation of DAPK1 inhibited IL-1β-induced inflammation and cartilage degradation via the PEDF-mediated NF-κB and NLRP3 inflammasome pathways.

The mechanism of CFB in treating knee osteoarthritis is not yet clear and deserves further discussion. The C28/I2 cell was stimulated by TNF-α and the MIA-induced OA rat model were constructed, and then treated with a certain concentration of CFB. The effects of CFB on chondrocyte apoptosis, inflammatory response, and collagen matrix degradation were assessed. Furthermore, we analyzed the regulation of CFB on the DDIT3 and Hedgehog pathways through western blot analysis. The smoothened agonist inhibitor SAG and DDIT3 overexpression lentivirus were applied to investigate how CFB regulates DDIT3 and Hedgehog pathway to protect against osteoarthritis. Our experimental results proved the protective of CFB against TNF-α stimulated C28/I2 cells. CFB treatment downregulated the DDIT3 protein and inactivated the HH pathway in TNF-α stimulated C28/I2 cells, and this effect may be related to the DDIT3 or HH pathway. Furthermore, the inhibitory effect of CFB on the HH pathway is related to DDIT3. In vivo animal assays showed that CFB can inhibit the degradation of cartilage collagen matrix, inhibit chondrocyte apoptosis, improve chondrocyte damage, and alleviate pain in arthritis rats, and the effect of CFB on OA rats is related to the HH pathway mediated by DDIT3. In summary, CFB has significant therapeutic effects on osteoarthritis, protecting cartilage degradation and damage, and inhibiting inflammatory responses. DDIT3 may participate as an intermediate molecule in the protective effect of the drug on OA. The SHH/GLi1 pathway is regulated by CFB through DDIT3.

Osteosarcoma (OS) is characterized by aggressive features including invasiveness and high incidence of metastasis. OS patients with metastases are difficult to treat and suffer from a poor prognosis. DPY30 (protein dpy-30 homolog) is a key component of SET1/MLL family of H3K4 methyltransferases, which is implicated in the progression of multiple cancers. However, the potential functional engagement of DPY30 in OS remains to be unveiled. The objective of this study is to investigate the potential roles of DPY30 in the regulation of malignant phenotypes of OS cells. We examined DPY30 expression from a published dataset (GSE28424) as well as in OS tissues and adjacent normal tissues from OS patients. The association of DPY30 expression level and clinicopathologic parameters was assessed by Chi-square test. The role of DPY30 in regulating the malignant phenotype of OS cells and tumorigenesis was examined by in vitro functional assays and xenograft mouse model. We reported an upregulation of DPY30 in OS tumor tissues in both published dataset and clinical samples. A high level of DPY30 expression was associated with larger tumor size and more metastasis in OS patients, as well as poor overall survival. DPY30 knockdown in OS cells significantly impairs proliferation, migration and invasion, but induced cellular apoptosis. We further demonstrated that the agonist of PI3K/AKT pathway can rescue the inhibitory effects of DPY30 knockdown in OS cells. Together, our data indicate that DPY30 functions as an oncogene to promote the malignancy of OS cells possibly through PI3K/AKT pathway. The dependency of OS cells on DPY30 overexpression is a targetable vulnerability in OS cells.

Histone modification H3K27me3 is an important chromatin mark that plays vital roles in repressing expression of developmental genes. Here, we construct high-resolution 3D genome maps using long-read chromatin interaction analysis by paired-end tag sequencing (ChIA-PET) and characterize H3K27me3-associated chromatin interactions in an elite rice hybrid, Shanyou 63. We find that many H3K27me3-marked regions may function as silencer-like regulatory elements. The silencer-like elements can come into proximity with distal target genes via forming chromatin loops in 3D space of the nuclei, regulating gene silencing and plant traits. Natural and induced deletion of silencers upregulate expression of distal connected genes. Furthermore, we identify extensive allele-specific chromatin loops. We find that genetic variations alter allelic chromatin topology, thus modulating allelic gene imprinting in rice hybrids. In conclusion, the characterization of silencer-like regulatory elements and haplotype-resolved chromatin interaction maps provide insights into the understanding of molecular mechanisms underlying allelic gene silencing and plant trait controlling.

To observe the effect of melatonin intervention on rat knee osteoarthritis (KOA) model and explore its mechanism. A total of 81 Sprague–Dawley (SD) rats were employed. Haematoxylin and eosin (H E) staining and safranin o-solid green staining were used to observe the changes of pathology in KOA, and inflammation factors in serum were detected by enzyme-linked immunosorbent assay (ELISA), type II collagen (Col-II) was detected by immunohistochemistry, chondrocyte apoptosis was detected by TdT-mediated dUTP nick-end labeling (TUNEL). The expression of matrix metalloproteinases (MMPs) and JAK2/STAT3 signaling were detected by western blot. Melatonin treatment ameliorated the histomorphology of knee joint in rats compared to the model group. The contents of TNF-α, IL-6, and IL-1β in serum were decreased after melatonin treatment. In addition, compared to the model group, the positive expression of Col-II increased, the chondrocyte apoptosis decreased after melatonin treatment. Interestingly, the expression levels of MMP3, MMP9, MMP13, p-JAK2 and p-STAT3 decreased (p < 0.05). Importantly, melatonin combined with AG490 is significantly ameliorates histomorphology of knee joint, reduced cartilage loss compared with melatonin treatment alone. Melatonin treatment can effectively diminish the cartilage injury. Its mechanism may be related to protect the articular cartilage by reducing the release of inflammatory factors, inhibit the expression of MMPs and JAK2/STAT3 signaling.

Context Isoorientin has many biological activities, including antioxidant, anti-inflammatory, antitumor. However, the effect of isoorientin on postmenopausal osteoporosis remains unclear. Objective To evaluate the effect of isoorientin on postmenopausal osteoporosis. Materials and methods Sprague-Dawley rats were divided into five groups (n = 5): sham, model, 17-beta-oestradiol (E2, 10 mu g/kg/day), low-dose isoorientin (L-Iso, 50 mg/kg), and high-dose isoorientin (H-Iso, 100 mg/kg). The rats were ovariectomized, treated by gavage daily for 12 weeks, and serum and femur samples were collected. Bone mineral density, bone metabolism, and oxidative stress were assessed. H&E staining, immunohistochemistry, and western blotting were employed. Results Isoorientin improved the bone mineral density of the lumbar vertebrae (2.01 +/- 0.05 g/cm(3) in H-Iso group vs. 1.74 +/- 0.07 g/cm(3) in model group) and femur (1.46 +/- 0.06 g/cm(3) vs. 1.19 +/- 0.03 g/cm(3)), increased the trabecular bone number (1.97 +/- 0.03 vs. 1.18 +/- 0.13) and thickness (0.27 +/- 0.02 vs. 0.16 +/- 0.03 mm). Isoorientin decreased the separation degree of trabecular bone, ameliorated bone histomorphology changes, and significantly improved the mechanical properties. Isoorientin diminished MDA (by 60%) and increased SOD (by 49.2%), and GSH-Px (by 159%) activity. Furthermore, osteoprotegerin (OPG), nuclear factor erythroid 2-like 2 (Nrf2), haem oxygenase (HO-1), NAD(P)H quinone dehydrogenase 1(NQO1), and oestrogen receptor 1(ESR1) protein expression increased, while receptor activator of nuclear factor-kappa B ligand (RANKL) protein expression decreased after treatment. Conclusions Isoorientin ameliorates osteoporosis via upregulating OPG and Nrf2/ARE signalling, suggesting isoorientin maybe a potential therapeutic drug for PMOP.

Purpose: To investigate the expressions of rapamycin target protein (mTOR) conduction pathway in human osteosarcoma MG-63 cells and their stem cells, and to examine the inhibitory effect of different doses of rapamycin. Methods: mTOR mRNA in osteosarcoma stem-like cells and human osteosarcoma MG-63 cells were determined by quantitative reverse transcription polymerase chain reaction (qRT-PCR). The cells were treated with different doses of rapamycin and divided into low dose group (0.5 mg), medium dose group (1.0 mg), high dose group (2.0 mg) and blank (control) group. Apoptosis and cell cycle of MG-63 cells were determined by flow cytometry, while proliferation of MG-63 cells up was assessed by CCK-8 kit. Results: mTOR in human osteosarcoma MG-63 cells was significantly lower than that in osteosarcoma stem-like cells. Compared with the control group, mRNA expression levels of mTOR in MG-63 cells and osteosarcoma stem-like cells were significantly decreased after treatment with different concentrations of rapamycin (p < 0.05). MG-63 cells treated with various doses of rapamycin exhibited a significant decrease in their proliferation, compared with control group, while only the high rapamycin concentration group exhibited a significant decrease in osteosarcoma stem-like cell proliferation (p < 0.05). Treatment with rapamycin in MG-63 cells and osteosarcoma stem-like cells resulted in a significant increase in apoptosis, prolonged G0/G1 phase and shortened S phase (p < 0.05). Conclusion: Rapamycin inhibits the expression of mTOR mRNA in osteosarcoma stem-like and MG-63 cells. It also inhibits the proliferation and cell cycle formation of osteosarcoma stem-like cells and MG-63 cells via mTOR signal pathway. These findings may provide a new target for the treatment of osteosarcoma.

Objective:To investigate the molecular mechanism of long intergenic non-protein-coding RNA 174 regulating osteoporosis.Methods:Osteoporosis patients and normal healthy people were selected. Human bone marrow mesenchymal stem cells (hBMSCs) were purchased from ScienCell company. Overexpression long intergenic non-protein coding ribonucleic acid 174 (LINC00174) plasmid (pcDNA-LINC00174), overexpression negative control (pcDNA-NC), LINC00174 small interfering RNA (si-LINC00174), and small interfering RNA negative control (si-NC) were transfected into hBMSCs. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to detect mRNA expression. Methylthiazolyldiphenyl-tetrazolium bromide (MTT) was used to detect cell proliferation activity. Cell apoptosis rate was detected by flow cytometry. The protein expression was detected by Western blotting. The malondialdehyde (MDA), superoxide dismutase (SOD) and catalase (MDA) were detected by enzyme-linked immunosorbent assay. The t test was used for comparison between two groups, the one-way analysis of variance was used for comparison between multiple groups, and the LSD- t test was used for pairwise comparison between groups. Results:The expression level of LINC00174 in serum of patients with osteoporosis was significantly higher than that of healthy controls (2.38±0.21 vs. 1.00±0.11, t=39.908, P<0.05). The expression of LINC00174 (3.01±0.25 vs. 1.00±0.08), apoptosis rate [(23.25±1.42)% vs. (12.46±1.07)%], cleaved cysteinyl aspartate specific proteinase-3 (0.75±0.05 vs. 0.39±0.03) in pcDNA-LINC00174 group was significantly higher than pcDNA-NC group, while proliferation activity (0.53±0.04 vs. 0.91±0.06), cyclin D1 (0.42±0.05 vs. 0.86±0.06), alkaline phosphatase (ALP) (0.37±0.03 vs. 0.65±0.05), runt-related transcription factor 2 (RUNX2) (0.41±0.04 vs. 0.83±0.06) and osteocalcin (OCN) (0.27±0.03 vs. 0.72±0.05) protein expression levels were significantly lower ( t=15.671, 20.877, 9.564, 12.431, 17.498, 21.652, 18.932, 21.774, P<0.05). The protein expression of nuclear factor erythroid 2-related factor 2 (Nrf2) (0.39±0.03 vs. 0.88±0.06) and Kelch-like ECH-associated protein 1 (Keap1) (0.24±0.02 vs. 0.59±0.05) in hBMSCs in pcDNA-LINC00174 group were significantly higher than the pcDNA-NC group ( t=17.238, 14.045, P<0.05). The protein expression levels of Nrf2 (0.94±0.05 vs. 0.72±0.04) and Keap1 (0.86±0.05 vs. 0.47±0.04) in hBMSCs in the si-LINC00174 group were significantly higher than those in the si-LINC00174 group ( t=13.032, 16.541, P<0.05). Conclusion:LINC00174 can regulate osteoporosis probably by regulating the proliferation, osteoblast differentiation, apoptosis and oxidative stress of hBMSCs through Nrf2/Keap1 signaling pathway.

Additional file 10: Table S9. RNA-Seq data of the different tissues used in this study.

We attempted to analyze the effects of cisplatin and exogenous hydrogen on the invasion and proliferation of human osteosarcoma cells through EGFR/P13K/Akt signaling pathway. The effects of exogenous hydrogen sulfide and cisplatin on the invasion, apoptosis and proliferation of MG-63 cells were detected by transwell chamber invasion experiment, flow cytometry and CCK-8 method. After 24 and 48 hours of drug action, the inhibition rate of cell proliferation was significantly higher than in the combined treatment. The number of cells decreased significantly, compared with the control group. Western blotting and qRT-PCR showed that the expression of Akt mRNA in MG-63 cells treated with cisplatin+hydrogen sulfide at different concentrations for 48 hours decreased. The expression of Akt, P13K and EGFR were decreased Flow cytometry analysis showed that after the drug treatment of MG-63 cells, the S phase of cisplatin and cisplatin+exogenous hydrogen concentrations with different prolonged concentrations was significantly shortened phase and the G0/G1 was significantly, and the apoptosis rate of each group was significantly increased. Cisplatin and exogenous hydrogen sulfide can synergistically inhibit the proliferation and invasion of human osteosarcoma MG-63 cells, and regulate the apoptosis mechanism of MG-63 cells through EGFR/P13K/Akt signaling pathway.

The complexity of the epigenome landscape and transcriptional regulation is significantly increased during plant polyploidization, which drives genome evolution and contributes to the increased adaptability to diverse environments. However, a comprehensive epigenome map of Brassica napus is still unavailable. In this study, we performed integrative analysis of five histone modifications, RNA polymerase II occupancy, DNA methylation, and transcriptomes in two B. napus lines (2063A and B409), and established global maps of regulatory elements, chromatin states, and their dynamics for the whole genome (including the An and Cn subgenomes) in four tissue types (young leaf, flower bud, silique, and root) of these two lines. Approximately 65.8% of the genome was annotated with different epigenomic signals. Compared with the Cn subgenome, the An subgenome possesses a higher level of active epigenetic marks and lower level of repressive epigenetic marks. Genes from subgenome-unique regions contribute to the major differences between the An and Cn subgenomes. Asymmetric histone modifications between homeologous gene pairs reflect their biased expression patterns. We identified a novel bivalent chromatin state (with H3K4me1 and H3K27me3) in B. napus that is associated with tissue-specific gene expression. Furthermore, we observed that different types of duplicated genes have discrepant patterns of histone modification and DNA methylation levels. Collectively, our findings provide a valuable epigenetic resource for allopolyploid plants.

Characterizing genome-wide histone posttranscriptional modifications and transcriptional factor occupancy is crucial for deciphering their biological functions. Chromatin immunoprecipitation followed by sequencing (ChIP-seq) is a powerful method for genome-wide profiling of histone modifications and transcriptional factor-binding sites. However, the current ChIP-seq experimental procedure in plants requires significant material and several days for completion. CUT&Tag is an alternative method of ChIP-seq for low-sample and single-cell epigenomic profiling using protein A-Tn5 transposase fusion proteins (PAT). In this study, we developed a nucleus CUT&Tag (nCUT&Tag) protocol based on the live-cell CUT&Tag technology. Our results indicate that nCUT&Tag could be used for histone modifications profiling in both monocot rice and dicot rapeseed using crosslinked or fresh tissues. In addition, both active and repressive histone marks such as H3K4me3 and H3K9me2 can be identified using our nCUT&Tag. More importantly, all the steps in nCUT&Tag can be finished in only 1 day, and the assay can be performed with as little as 0.01 g of plant tissue as starting materials. Therefore, our results demonstrate that nCUT&Tag is an efficient alternative strategy for plant epigenomic studies.

Objective: This study aimed to investigate the possible relationship between Scheuermann disease (SD) and the pathophysiological factors of thoracic spinal stenosis (TSS), including ossification of the ligamentum flavum (OLF), ossification of the posterior longitudinal ligament (OPLL), and thoracic disc herniation (TDH) in patients with symptomatic TSS. Methods: Demographic and radiological data from 66 consecutive patients diagnosed with symptomatic TSS from 2013 to 2018 were retrospectively collected and divided into 3 groups depending on the underlying pathomechanism of TSS: TDH group (18 patients; 6 women; mean age +/- standard deviation [Sd] = 59.89 +/- 11.34), OPLL group (12 patients; 8 women; mean age +/- Sd = 56.08 +/- 14.74), and OLF group (36 patients; 20 women; mean age +/- Sd = 58.69 +/- 9.77). A total of 41 age-matched healthy individuals (19 women; mean age +/- Sd = 54.88 +/- 13.63) were designated as the control group. In each group, both typical and atypical SD criteria were radiologically examined. The demographic data and presence of SD between the control group and 3 subgroups of TSS pathomechanisms were evaluated. Results: SD characteristics were identified in 83.33% (15/18) of patients in the TDH group, 44.44% (16/36) in the OLF group, 25% (3/12) in the OPLL group, and 17.07% (7/41) of the control individuals. When analyzed by the chi-squared test and logistic regression analysis, the presence of SD was significantly associated with TDH (P < 0.01) and OLF (P < 0.05) but not OPLL (P > 0.05). Patients with TDH and OLF showed peak involvement of T10/11, and patients with OPLL did not. Furthermore, we determined that age, sex, body-mass index, and smoking status were not the risk factors for TDH, OPLL, and OLF (P > 0.05). SD was found to be a risk factor for TDH (P < 0.01) and OLF (P < 0.05) but not for OPLL (P > 0.05). Conclusion: Evidence from this study indicated that SD might be a risk factor for OLF and TDH but not for OPLL.