Background: Rotator cuff injuries are common musculoskeletal disorders and are frequently complicated by impaired tendon–bone healing and high re-tear rates after surgical repair. Exosomes derived from adipose-derived stem cells (ADSCs) have shown regenerative potential through paracrine mechanisms; however, the role of exosomal insulin-like growth factor 1 (IGF1) in tendon–bone healing remains unclear. Methods: Exosomes were isolated from rat ADSCs with or without lentiviral knockdown of IGF1. A rat supraspinatus tendon tear and repair model was established, and 200 μg of exosomes was administered systemically post-surgery. Tendon–bone healing was evaluated at 8 weeks post-operation using histological, immunohistochemical, and micro-computed tomography analyses. Early molecular responses were assessed at 1-week post-surgery by Western blot and RT-qPCR. Angiogenic markers (vascular endothelial growth factor (VEGF), CD31, α-SMA), inflammatory cytokines (interleukin (IL)-1β, IL-18), pyroptosis-related proteins (gasdermin D N-terminal fragment (GSDMD-N)), and NLRP3 inflammasome components were examined. Results: ADSC-derived exosomes significantly enhanced bone mineral density, fibrocartilage formation, vascularization, and biomechanical strength at the tendon–bone interface. These effects were accompanied by reduced inflammatory cytokine expression, inhibition of pyroptosis, and suppression of NLRP3 inflammasome activation. In contrast, exosomes derived from IGF1-deficient ADSCs exhibited markedly reduced therapeutic efficacy, with attenuated angiogenic, anti-inflammatory, and anti-pyroptotic effects. Conclusions: Exosomal IGF1 plays a critical role in promoting angiogenesis, suppressing inflammation and pyroptosis, and improving structural and biomechanical outcomes during tendon–bone healing. IGF1-enriched ADSC-derived exosomes represent a promising therapeutic strategy for enhancing rotator cuff repair.

In the field of bone defect repair, the three-dimensional architecture and bioactivity of tissue-engineered bone scaffolds play a pivotal role. However, two major challenges remain: the cell type–specific response of host cells to scaffold pore architecture and the sustained promotion of angiogenesis and osteogenesis essential for large-segment bone regeneration. Inspired by the structural characteristics of natural honeycombs, this study integrates 3D printing with hydrogel engineering to construct a novel composite scaffold. Its macro-level innovation lies in a multilevel pore structure, while its functional core stems from a meticulously designed “nano-bio interface”: layered double hydroxides (LDHs) loaded with deferoxamine (DFO) are assembled into nanosheets (DFO@LDHs) as key functional units, and are simultaneously integrated into both a polycaprolactone (PCL) framework (D@LP) and a glycidyl methacrylate-modified hyaluronic acid (HA-GMA) hydrogel (D@LG) to form the final composite scaffold (D@LG/D@LP). This nano-bio interface not only enables sustained, controlled release of bioactive factors but also actively regulates cellular spatiotemporal behavior. The unique pore distribution and degradation characteristics of the hydrogel and 3D-printed scaffold preferentially guide endothelial cell adhesion and microvascular network formation, subsequently initiating stem cell differentiation and bone matrix deposition. Both in vitro and in vivo experiments confirm the scaffold’s excellent biocompatibility. Through a nanomaterial-driven cascade regulation mechanism, it coordinates the activation of angiogenesis and osteogenesis, significantly accelerating vascularized bone regeneration. This work demonstrates an effective strategy for empowering three-dimensional scaffolds through nano-bio interface construction, achieving spatiotemporal programming for complex tissue regeneration.

OBJECTIVE:High fasting plasma glucose (HFPG) is a significant risk factor for lower extremity peripheral arterial disease (PAD). No studies have investigated the detailed global burden of lower extremity PAD associated with HFPG. It is important to evaluate the current epidemiology and changing trends of HFPG related lower extremity PAD. METHODS:This study extracted two main indicators from the Global Burden of Disease database, including deaths and disability adjusted life years (DALYs) of lower extremity PAD associated with HFPG from 1990 to 2021. Subanalyses of geographic distribution, sex, age, and sociodemographic index (SDI) were performed, and estimated annual percentage changes were calculated to provide detailed insights into the disease burden. RESULTS:In 2021, there were 25 640 deaths and 562 161 DALYs from lower extremity PAD associated with HFPG worldwide. The global age standardised rates (ASRs) per 100 000 population of deaths and DALYs were 0.32 and 6.73, respectively. The rates of both deaths and DALYs increased with advancing age, peaking in the 95+ years age group, with rates of 31.509 and 292.935 per 100 000, respectively. The rates of deaths and DALYs were, respectively, 0.335 and 7.165 for males and 0.315 and 7.082 for females in 2021. From 1990 to 2021, the ASRs of deaths and DALYs associated with HFPG continued to increase, surpassing those associated with smoking, high body mass index, high systolic blood pressure, and kidney dysfunction. CONCLUSION:Lower extremity PAD associated with HFPG remains a significant public health concern, with an uneven geographic distribution. The disease burden is higher in high SDI countries and among elderly individuals. Additionally, greater attention should be directed towards females and low and middle income countries, as the burden of PAD associated with HFPG is gradually increasing in these groups and countries.

Aortic aneurysm (AA) is a life-threatening arterial disease. It is imperative to understand the epidemiology of AA and implement effective interventions. Publicly available data on deaths and years of life lost (YLLs) from AA between 1990 and 2021 were retrieved from the Global Burden of Disease study 2021. Counts and rates for deaths and YLLs were presented, along with their corresponding 95% uncertainty intervals. Estimated annual percentage changes were calculated to reflect the temporal trends in the burden of AA. A Bayesian age-period-cohort model was used to generate projections to 2050. The global numbers of deaths and YLLs due to AA have increased from 1990 to 2021. In 2021, 153 927 deaths and 3 107 762 YLLs were attributed to AA. In contrast, the global age-standardized rates (ASRs) of AA-related deaths and YLLs have decreased from 1990 to 2021, with rates of 1.86 and 36.54 per 100 000 population in 2021, respectively. The decline in ASRs of deaths and YLLs was projected to continue until 2050. The burden of AA decreased from 1990 to 2021 in regions and countries with a high socio-demographic index but increased in less developed regions and countries. In 2021, male gender, smoking, and high systolic blood pressure were the primary risk factors contributing to AA. AA remains a major public health concern, especially among the elderly, smokers, and men in less developed countries. Smoking cessation and hypertension control appear to be effective in reducing the burden of AA.

The spatiotemporal knockdown of genes through genome editing heralds a new frontier in molecular breeding,yet it remains largely unexplored.Recognizing the intricate regulatory networks of endogenous microRNAs(miRNAs),we posited that integration of specific miRNA target sequences into the 3'untrans-lated region(UTR)of a gene could construct artificial miRNA-dependent regulatory circuits,facilitating pre-cise spatiotemporal gene suppression.To test this hypothesis,we selected three endogenous miRNAs with unique expression profiles by analyzing rice miRNA expression profiles.Results from both transient assays and stably-edited rice plants confirmed that in-locus incorporation of miRNA targets into the 3'UTR of target genes can substantially reduce their expression in a spatiotemporal manner.Using GID1 as a target gene,we found that knockin of the miR156a target led to a remarkable 97％constitutive reduc-tion;knockin of the tissue-specifically expressed miR396c target significantly reduced its expression in shoots alone;and knock-in of the long-day-induced miR528 target triggered a dramatic and temporal decrease of 95％specifically under such light exposure.These findings underscore the viability of miRNA-mediated,in-locus knockdown(MiRKD)as a convenient approach for crop breeding,leveraging miRNA expression traits and genome editing for conditional gene suppression.

Purpose: This study aimed to assess the safety and viability of combining branched stent graft with fenestrated thoracic endovascular aortic repair (TEVAR) in treating aortic arch lesions. Materials and Methods: The cohort included patients presenting with aortic arch lesions who underwent treatment with a combination of branched stent graft and fenestrated TEVAR between July 2020 and November 2022. Technical success was defined as the precise deployment of the stent graft, maintenance of branch vessel patency, and the absence of type I endoleak. The secondary outcomes examined were complications and all-cause mortality. Results: The study cohort comprised 21 patients (average age: 61.0±14.8 years) with aortic arch lesions from 3 tertiary care hospitals. The aortic arch lesions encompassed aortic dissection (N=8), aortic aneurysm (N=8), pseudoaneurysm (N=1), intramural hematoma (N=1), and penetrating aortic ulcer (N=3). The technical success rate achieved was 95.2% (20/21). Failure in one case was due to an intraoperative type I endoleak, which was rectified with an additional stent graft placement. The 30-day mortality rate was 4.8% (1/21). One patient suffered a stroke but responded well to medical intervention. The median hospital stay was 10.9±5.4 days. During the follow-up period, one death (4.8%) was associated with aortic complications. A type II endoleak was observed and managed with close monitoring. Two patients underwent re-interventions for retrograde type A dissection and stent migration, respectively. No occlusions were observed in the target branch arteries. Conclusions: The combination of branched stent graft with fenestrated TEVAR emerges as a viable strategy for addressing specific lesions in the aortic arch. Clinical Impact This study demonstrates the feasibility of using branched stent grafts with fenestrated TEVAR for treating aortic arch lesions, achieving a technical success rate of 95.2%. Compared to traditional open surgery, this innovative, minimally invasive approach reduces perioperative mortality and complications, such as stroke and spinal cord ischemia. For clinicians, it offers a viable alternative for patients unfit for open repair, particularly in complex aortic arch cases. While the initial outcomes are promising, further research is needed to assess long-term durability and risks, including stent graft migration and late endoleak, ensuring the technique’s safety and efficacy over time.

Abstract Background: Infected aortic aneurysm is one of the most life-threatening cardiovascular diseases. There is still no standardized and uniform practice in the management of it. Objective: This study aimed to comprehensively analyze all the literatures related to infected aortic aneurysm in the past decades by using bibliometric tools from Web of Science Core Collection, revealing the research dynamics and hotspots in this field. Methods: We summarized the articles in the Web of Science Core Collection for infected aortic aneurysm in each year from January 2000 to October 2023. The bibliometric tools including SCImago, VOSviewer and CiteSpace were used to perform data extraction and visualization. The national characteristics of macro-geographical distribution, the contributions of disciplines, journals and authors, and cooperation network of institutions and authors were revealed. Results: A total of 1088 publications were included. The distribution of publications by year showed that the number of publications rose steadily since2004, followed by a decline from 2013 to 2017 and reaching a peak in 2021.The geographical distribution indicated that there was a large gap between the number of publications and citations in the United States and other countries.The co-authorship analysis showed that collaboration between prolific countries, productive institutions or excellent authors was scarce. The infected aortic aneurysm research was a multi-disciplinary field and most of the articles were published in journals related to surgery including Journal of Vascular Surgery, European Journal of Vascular and Endovascular Surgery etc. The keyword analysis revealed clustering and burst terms, indicating that current research focused on treatment of infected aortic aneurysm. Conclusions: This study revealed trends in research and public interest in infected aortic aneurysm. Our findings suggested that the field was growing, and the United States taking the lead position. Moreover, cooperation between countries should be strengthened, and future research hot spots might focus on deeper exploration of drug mechanisms, prediction of treatment efficacy, prediction of adverse events, and new modes of administration, such as combination therapy, which may pave the way for further research.

Base editors, including cytosine and adenine base editors (CBE and ABE), are promising tools for precise genome modification. They enable the generation of single nucleotide variants in plants for research and crop improvement (Li et al., 2020; Manghwar et al., 2019; Ren et al., 2021; Xu et al., 2021; Zeng et al., 2022). However, existing base editors are still limited in the types of base conversions they can induce. Recently, a new base editor was constructed by fusing an engineered N-methylpurine DNA glycosylase (MPG) with ABE to create AYBE. This has achieved efficient A-to-T and A-to-C (A-to-Y, AYBE) transversions in mammalian cells and also timely assessed in rice to induce A-to-T (AKBE) (Li et al., 2023; Tong et al., 2023; Wu et al., 2023). However, the editing activity of AYBE remains unexplored in maize, and its editing efficiency leaves room for further optimization. Here, by fusing the adenine base editor with a codon-optimized N-methylpurine DNA glycosylase (MPG) and co-expressing the maize translesion synthesis DNA polymerase η (Polη), we developed an optimized AYBE base editor (ZmAYBEv3) for both A-to-T and A-to-C base conversions with high efficiency in maize and other monocots plants. First, the human-derived MPG (hMPG) was engineered (G163R, N169S, S198A, K202A, G203A, S206A and K210A) and codon-optimized for maize (MzMPG), then fused to the C-terminus of the maize ABE editor ZmABE8e to construct the initial AYBE editor ZmAYBEv1 (Figure 1a). Two sgRNAs (sgRNA1 and sgRNA2) targeting maize genes ZmGA20ox3 and ZmCT2 were designed. Hundreds of young embryos from the inbred maize variety KN5585 were transformed with Agrobacterium for evaluation. Approximately 50 regenerated shoots from each transformation were pooled and genotyped using the next-generation sequencing (NGS). As expected, only A-to-G substitutions were detected in samples edited with the conventional ZmABE8e, while A-to-Y conversions were found in ZmAYBEv1 edited samples (Figure 1b). For example, at the A8 site of sgRNA1, the A-to-T and A-to-C conversion frequencies were 3.86% and 0.53%, respectively, demonstrating the A-to-Y editing activity of ZmAYBEv1. We then tested it in maize plants. A total of 45 T0 plants were obtained and genotyped by NGS (Liu et al., 2019). The results showed seven T0 plants contained A-to-Y substitutions, further demonstrating ZmAYBEv1's editing capability in plantlet (Figure 1c; Table S1). However, the chimerism state of A-to-Y substitutions (calculated from the proportion of NGS reads, Li et al., 2023) was too low in most mutants. Usually, T0 plants with a chimerism>10% are required to ensure heritability. Thus, only one mutant could be identified as a valid A-to-T edited line, and no A-to-C editing lines were found. This revealed the need for further improvement of ZmAYBEv1. Polη is involved in the replication of damaged DNA and may improve base editing efficiency (Tong et al., 2023). Accordingly, human and maize Polη (hPolη and ZmPolη) were incorporated into ZmAYBEv1 to construct ZmAYBEv2 and ZmAYBEv3, respectively (Figure 1a). Quick tests in maize embryos showed a significant increase in A-to-T and A-to-C editing efficiencies when using ZmAYBEv3 (Figure 1b), indicating positive regulation of ZmPolη on AYBE. We think that different base conversion types between pAYBEv2 and pAYBEv3 might be caused by the different enzymatic activity of hPolη and ZmPolη (Figure S1). To assess them in transgenic plants, 39 and 52 T0 plants were generated using ZmAYBEv2 and ZmAYBEv3, respectively, targeting the same two genes above (Figure 1c). As expected, substantially more A-to-Y edited plants (7 out of 23 with chimerism>10%, the same hereinafter) were identified in ZmAYBEv3 edited lines for ZmGA20ox3. At sgRNA2 of ZmCT2, an uneditable site for ZmAYBEv1 or ZmAYBEv2, an A-to-T editing plant was successfully obtained using ZmAYBEv3. Moreover, three A-to-C edited lines were also identified. To confirm the editing results, we then resequenced the ZmAYBEv3-derived lines by Sanger sequencing and further confirmed these results (Figure 1d; Figure S2). Notably, we also found that homozygous lines could be generated in T0 plants. The homozygous A-to-T editing at the sgRNA1 (A8) of ZmGA20ox3 produced a premature stop codon (AAG to TAG), resulting in a semi-dwarf phenotype of maize, even in T0 generation (Figure 1e,f). These results indicate ZmAYBEv3 has the highest editing efficiency, capable of both A-to-T and A-to-C editing. To further confirm the versatility of ZmAYBEv3, we targeted three additional maize genes (ZmLW2, ZmABH2 and ZmLBD5) for editing. We regenerated 51 T0 maize plants and performed NGS genotyping. The results showed successful A-to-Y editing by ZmAYBEv3 at all three genes, with an average efficiency of 35.3% (18/51). Notably, the A-to-T editing frequency reached 45.5% at the ZmLBD5 locus. Given the known transferability of base editors across monocot species, we also tested ZmAYBEv3 in rice on three genes (OskTN80b, OsWaxy and OsTB1). NGS and Sanger sequencing showed ZmAYBEv3 could efficiently induce A-to-Y editing at the three rice genes with an average efficiency of 21.1% (12/57) (Figure 1c,d; Figure S3). In some locus, the A8 site within a sgRNA seems the best targeting nucleotide (Figure 1g). Together, these results further validate the editing activity of ZmAYBEv3 in both maize and other monocot species. Collectively, the incorporation of ZmPolη enhanced the A-to-Y editing efficiency of ZmAYBEv3. Across five target sites in 103 T0 maize plants, 50 plants had A-to-Y conversions (chimerism >1%), validating its capabilities. Notably, 26 plants (25.2%) showed potentially heritable edits (chimerism>10%). ZmAYBEv3 also enables the possibility of obtaining homozygous edits within the T0 generation. The high editing efficiencies achieved by ZmAYBEv3 in maize and rice highlight its usefulness as an alternative tool to supplement existing base and prime editors for functional studies and trait improvement in crops. Supported by the National Key R&D Program of China (No. 2021YFD1201300) and the National Natural Science Foundation of China (No. 32070396) to Y.L. We thank WIMI for assistance with maize transformation. D.Z. and Y.L. designed the research; D.Z, H.P., K.L., Y.Z., F.Z., L.Y., S.R., Q.D. and J.X. performed experiments; D.Z. and Y.L. wrote and revised the manuscript. The authors declare no competing interests. The data that supports the findings of this study are available in the supplementary material of this article. Data S1 Supplemental methods. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

Pristimerin (PM), serving as a biological component mainly obtained from Celastraceae and Hippocrateaceae families, has been extensively explored for its numerous pharmacological activities, especially anti-cancer activity. However, the function of PM on pathological cardiac hypertrophy is poorly understood. This work was intended to investigate the effects of PM on pressure-overload induced myocardial hypertrophy and its potential pathways. Mouse model of pathological cardiac hypertrophy was generated by transverse aortic constriction (TAC) or minipump administration of the beta-adrenergic agonist ISO for 4 weeks, and PM (0.5 mg/Kg/d, i.p.) was treated for 2 weeks. PPAR alpha-/- mice received TAC surgery were used for mechanism exploration. Moreover, neonatal rat cardiomyocytes (NRCMs) were utilized to explore the effect of PM following Angiotensin II (Ang II, 1.0 mu M) administration. We found that PM attenuated pressure-overload induced cardiac dysfunction, myocardial hypertrophy and fibrosis in mice. Likewise, PM incubation dramatically reversed Ang II-mediated cardiomyocytes hypertrophy in NRCMs. RNA-Sequence showed that PM selectively contributed to improvement of PPAR alpha/PGC1 signaling, while silencing PPAR alpha abrogated the beneficial effects of PM on Ang II-treated NRCMs. Importantly, PM ameliorated Ang II-induced mitochondrial dysfunction and decrease in metabolic genes, whereas knockdown of PPAR alpha eliminated these alterations in NRCMs. Similarly, PM presented limited protective effects on pressure-overload induced systolic dysfunction and myocardial hypertrophy in PPAR alpha deficient mice. Overall, this study revealed that PM exerted protective activity against pathological cardiac hypertrophy through improvement of PPAR alpha/PGC1 pathway.

Objective:The aim of this study is to evaluate the efficacy of endovascular treatment for nondissected diseases of the ascending aorta. Data Sources. PubMed, Embase, and SciELO. Review Methods. In this study, we conducted a search on the PubMed, Embase, and SciELO databases for all cases of ascending aortic endovascular repair included in the literature published between January 2007 and July 2023, excluding type A aortic dissection. We reviewed 56 case reports and 7 observational studies included in this study, assessing the techniques, equipment, procedural steps, and results. We summarized the age, complications, follow-up time, and access route.Results:This study includes 63 articles reporting 105 patients (mean age: 64.96 ± 17.08 years) who received endovascular repair for nondissected ascending aortic disease. The types of disease include aneurysm (N = 16), pseudoaneurysm (N = 71), penetrating aortic ulcer (N = 10), intramural hematoma (N = 2), thrombosis (N = 2), iatrogenic coarctation (N = 1), and rupture of the aorta (N = 3). The success rate of surgery is 99.05% (104/105). Complications include endoleak (10.48%, 11/105), stroke (5.71%, 6/105), postoperative infection (1.91%, 2/105), acute renal failure (0.95%, 1/105), aortic rupture (0.95%, 1/105), thrombosis (0.95%, 1/105), and splenic infarction (0.95%, 1/105). Five patients required conversion to open surgery, two patients underwent endovascular reintervention, and four of these five patients underwent surgery due to endoleak. Early mortality was 2.86% (3/105).Conclusion:While the viability and results of endovascular repair for the treatment of ascending aortic disease are acknowledged in some circumstances, further research is needed to determine the safety and effectiveness of endovascular treatment for ascending aortic disease.

Nowadays, many studies focus on the preparation of small-diameter vascular scaffolds, but some issues still existed that need to be urgently solved, but there are still problems such as slow endothelialization and failure to keep the vessels open for a long time that need to be solved. In this paper, a composite process of preparing triple-layer vascular scaffolds with microchannel structure in the inner layer is proposed to guide cell growth and accelerate the endothelialization process. The PCL pattern was printed as the intermediate layer of the vascular scaffold by 3D printing method, and the width of the microchannel in the inner layer of the scaffold was controlled by the printing spacing. The inner and outer layers were prepared by electrospinning. Heparin is often used as an anticoagulant in vascular tissue engineering to improve the problem of thrombosis and blockage of vascular scaffold after transplantation into the body. Here, the inner layer carries heparin immobilized with silk fibroin by coaxial electrospinning, so that heparin can be released slowly. The addition of silk fibroin also improved the hydrophilicity of PCL electrospinning film by water contact angle test. The mechanical properties of vascular scaffolds with double intermediate layers (DIL) are better than those with single intermediate layers (SIL) and also better than pure electrospinning scaffolds. In vitro cell experiments showed that cells grow directionally on microchannel structures of inner layer and randomly on electrospinning film without microchannel structure. The results showed that the microchannels were more conducive to cell growth and proliferation at a diameter of 0.25 mm. The composite electrospinning films with silk fibroin and heparin facilitated cell adhesion and proliferation compared to the pure PCL films. All these results indicate that the vascular scaffolds have potential applications in clinic for vascular tissue regeneration.

Osteoarthritis is a common degenerative joint disease that can cause pain and disability in patients. There is still a lack of effective treatments to improve pathological changes of osteoarthritis cartilages and reverse the progression of osteoarthritis. Our study aimed to investigate the role of Dlx5 in papain-induced osteoarthritis. Osteoarthritis was induced through intraarticular injection of papain. The pathological damage of cartilage tissues was analyzed by H&E staining. The apoptosis of cartilage tissues was detected by TUNEL assay. Immunohistochemical staining was performed to detect DLX5 and BMP-2. Western blot was performed to detect the expressions of SP7, caspase-3, and MYC. The results showed that administration of anti-Dlx5 improved pathological changes of osteoarthritis cartilages, characterized by decreased chondrocyte proliferation, chondrocyte hypertrophy, and matrix damage. Anti-Dlx5 treatment decreased the expressions of BMP-2 and SP7, which are positive regulators of chondrocyte hypertrophy. Moreover, MYC and caspase-3, the critical mediators for chondrocyte apoptosis, were both decreased after anti-Dlx5 treatment. In conclusion, anti-Dlx5 retarded the progression of osteoarthritis by downregulating chondrocyte hypertrophy and chondrocyte apoptosis-related genes. Our findings suggests that Dlx5 is a promising target for osteoarthritis treatment.

Sirtuin1 (Sirt1)/forkhead box O1 (FoxO1) axis has been reported as a crucial regulator involved in chondral homeostasis of healthy or osteoarthritis (OA) cartilage. In our study, the aim is to investigate whether dioscin functions as an activator of Sirt1/FoxO1 to protect against mechanical stress-induced chondrocyte dysfunction in vitro and in vivo models. HERB and PubChem databases were implemented to predict dioscin-related gene targets. Cell and mouse models of OA were established to determine the pharmacological value of dioscin, a steroidal saponin. Cartilage loss in the knee joint was detected by Safranin O staining. Phosphorylation and nucleocytoplasmic shuttling of FoxO1 was observed in mechanical stress-stimulated chondrocyte and anterior cruciate ligament transection-induced cartilage injury. However, dioscin treatment repressed FoxO1 phosphorylation and cytoplasmic transfer and elevated Sirt1 protein expression. Dioscin treatment reversed mechanical stress-induced growth inhibition and apoptosis of chondrocytes and improved cartilage degradation and bone loss in the epiphysis of the distal femur. Moreover, dioscin could maintain the normal phenotype of chondrocytes via mediating multiple gene expressions. Dioscin inhibited apoptosis and metabolic disorders in OA-like chondrocytes via maintaining the transcriptional activity of FoxO1 and enhancing Sirt1 expression. Dioscin might be a potential Sirt1 activator providing a novel therapeutic schedule for the treatment of OA.

ABSTR A C T Background: During the pathogenesis of tendinopathy, the chronic inflammation caused by the injury and apoptosis leads to the generation of scars. Ginsenoside Rg1 (Rg1) is extracted from ginseng and has anti-inflammatory effects. Rg1 is a unique phytoestrogen that can activate the estrogen response element. This research aimed to explore whether Rg1 can function in the process of tendon repair through the estrogen receptor. Methods: In this research, the effects of Rg1 were evaluated in tenocytes and in a rat model of Achilles tendinitis (AT). Protein levels were shown by western blotting. qRT-PCR was employed for evaluating mRNA levels. Cell proliferation was evaluated through EdU assay and cell migration was evaluated by transwell assay and scratch test assay. Results: Rg1 up-regulated the expression of matrix-related factors and function of tendon in AT rat model. Rg1 reduced early inflammatory response and apoptosis in the tendon tissue of AT rat model. Rg1 promoted tenocyte migration and proliferation. The effects of Rg1 on tenocytes were inhibited by ICI182780. Rg1 activates the insulin-like growth factor-I receptor (IGF1R) and MAPK signaling pathway. Conclusion: Rg1 promotes injured tendon healing in AT rat model through IGF1R and MAPK signaling pathway activation. ?? 2021 The Korean Society of Ginseng. Publishing services by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

To explore the molecular mechanism of cartilage calcification induced by osteoarthritis (OA) based on distal-less homeobox gene 5 - alkaline phosphatase - integrin-binding sialoprotein - ecto-nucleotide pyrophosphatase 1 (DLX5-ALPL-IBSP-ENPP1) signal axis. Twenty-four rabbits were selected to build models of cartilage calcification induced by OA and randomly divided into 3 groups. The first group was the normal group whose rabbits were injected into 0.9% saline (0.3 mL), and the second group was model group. The third group was model group whose rabbits were injected into DLX5 antibody by caudal vein. Alizarin red calcium staining was used to analyze calcium deposition of cartilage matrix. Immunohistochemical staining was used to analyze the relative expression levels of proteins DLX5 and ENPP1, and western blot was used to analyze the DLX5, ALPL, IBSP, and ENPP1 expression. Calcium salt precipitation was the most serious, and the calcification area increased in the model group. Although calcified nodules appeared in the anti-DLX5 group, they were relatively few. Immunohistochemical staining analysis showed that the protein DLX5 located in the nucleus and the protein ENPP1 located in the extracellular matrix. Western blot analysis showed that the expressions of proteins DLX5, ALPL, IBSP, and ENPP1 were the highest in OA Model group than that of NC group, followed by anti-DLX5 group. The proteins DLX5, ALPL, IBSP, and ENPP1 can promote cartilage calcification induced by OA based on DLX5-ALPL-IBSP-ENPP1 signal axis.

Objective: CircCCDC66 is involved in cancer progression, but its role in osteoarthritis (OA) remains unknown. This study was carried out to explore the biological role of circCCDC66 in OA and its underlying mechanism. Methods: The expression levels of miR-3622b-5p and circCCDC66 in OA cartilage tissues were detected by qRT-PCR. Cell Counting Kit-8 (CCK8) and flow cytometry were used to detect the chondrocyte viability and apoptosis. The expression of chondrocyte inflammatory factors (IL-6 and TNF-α) was measured by ELISA. The target genes of circCCDC66 and miR-3622b-5p were analyzed by bioinformatics analysis and luciferase reporter gene assay. The relationship between circCCDC66 and miR-3622b-5p was analyzed by bioinformatics analysis and luciferase reporter gene assay. Results: It was found that circCCDC66 expression in OA cartilage tissues was upregulated. CircCCDC66 overexpression inhibited proliferation and promoted apoptosis of chondrocytes and increased IL-6 and TNF-α levels in chondrocytes. miR-3622b-5p was predicted to be a downstream target gene of circCCDC66, and circCCDC66 overexpression inhibited miR-3622b-5p expression in chondrocytes. Moreover, miR-3622b-5p expression was downregulated in OA cartilage tissues. miR-3622b-5p overexpression increased chondrocyte proliferation, inhibited chondrocyte apoptosis, and enhanced the expression of IL-6 and TNF-α in chondrocytes. In addition, circCCDC66 overexpression enhanced SIRT3 expression in chondrocytes, while miR-3622b-5p overexpression inhibited SIRT3 expression in chondrocytes. Conclusion: CircCCDC66 promoted OA chondrocyte apoptosis by regulating the miR-3622b-5p/SIRT3 axis. CircCCDC66 may be a new therapeutic target of OA.

Miao, H.1; Cui, Y.1; Lu, Y.1; Sun, T.1; Dou, J.1; Ren, Y.1; Wang, C.2; Zhang, Y.3 Author Information

Aging is an important risk factor for osteoarthritis (OA). Butorphanol is a preoperative sedative and analgesic that possesses anti-inflammatory activity. However, the effect of butorphanol on OA has not been reported. Here we aimed to explore the effect of butorphanol tartrate on the cellular senescence of human chondrocyte-articular (HC-A) cells in response to tumor necrosis factor-α (TNF-α) stimulation. Butorphanol tartrate attenuated the TNF-α-caused cellular senescence of HC-A cells, with decreased positive senescence-associated-β-galactosidase (SA-β-gal) staining and elevated telomerase activity. Butorphanol tartrate prevented TNF-α-caused cell cycle arrest in the G0/G1 phase in HC-A cells and decreased p21 expression. The TNF-α-induced production of interleukin (IL)-6 and IL-8 in HC-A cells were mitigated by butorphanol tartrate. In addition, butorphanol tartrate reduced p-NF-κB p65/total p65 and p-STAT3/STAT3 ratios in HC-A cells cultured with TNF-α. Taken together, butorphanol tartrate protected HC-A cells from TNF-α-caused cellular senescence through inactivation of NF-κB and STAT3. These results imply that butorphanol tartrate might be used as a potential agent for the treatment of aging-related OA.

Gallbladder cancer (GBC) is the most common biliary tract malignancy worldwide. Although a growing number of studies have explored the mechanism of GBC, thus far, few molecules have been discovered that can be utilized as specific biomarkers for the early diagnosis and therapeutic treatment of GBC. Recent studies have shown that exosomes not only participate in the progression of tumors, but also carry specific information that can define multiple cancer types. The present study investigated the expression profiles of coding (or messenger) ribonucleic acids (mRNAs) and non-coding RNAs (ncRNAs, including long non-coding RNAs [lncRNAs] and circular RNAs [circRNAs]) in plasma-derived exosomes from GBC patients. Using high-throughput RNA sequencing and subsequent bioinformatic analysis, a number of differentially expressed (DE) mRNAs, lncRNAs, and circRNAs were identified in GBC exosomes, compared to their expressions in xantho-granulomatous cholecystitis (XGC) exosomes. Gene Ontology (GO) and Kyoto Encyclopedia of Gene and Genome (KEGG) analyses were then conducted to investigate the potential functions of these DE RNAs. Furthermore, the interaction networks and competing endogenous RNA networks of these DE RNAs and their target genes were investigated, revealing a complex regulatory network among mRNAs and ncRNAs. In summary, this study demonstrates the diagnostic value of plasma-derived exosomes in GBC and provides a new perspective on the mechanism of GBC.

Osteomyelitis is commonly caused by Staphylococcus aureus. Both erythromycin and curcumin can suppress S. aureus growth, but their roles in osteomyelitis are barely studied. We aim to explore the activities of erythromycin and curcumin against chronical osteomyelitis induced by methicillin-resistant S. aureus (MRSA). Chronicle implant-induced osteomyelitis was established by MRSA infection in male Wistar rats. Four weeks after bacterial inoculation, rats received no treatment, erythromycin monotherapy, curcumin monotherapy, or erythromycin plus curcumin twice daily for 2 weeks. Bacterial levels, bone infection status, inflammatory signals and side effects were evaluated. Rats tolerated all treatments well, with no death or side effects such as, diarrhea and weight loss. Two days after treatment completion, erythromycin monotherapy did not suppress bacterial growth and had no effect in bone infection, although it reduced serum pro-inflammatory cytokines tumor necrosis factor (TNF)-α and interleukin (IL)-6. Curcumin monotherapy slightly suppressed bacterial growth, alleviated bone infection and reduced TNF-α and IL-6. Erythromycin and curcumin combined treatment markedly suppressed bacterial growth, substantially alleviated bone infection and reduced TNF-α and IL-6. Combination of erythromycin and curcumin lead a much stronger efficiency against MRSA induced osteomyelitis in rats than monotherapy. Our study suggests that erythromycin and curcumin could be a new combination for treating MRSA induced osteomyelitis.