Repairing spinal cord injury (SCI) remains hindered by two major unmet challenges: the inability to precisely reconstruct irregular lesion cavities, where tissue contraction and collapse disrupt anatomical continuity, and the lack of effective platforms for stable and localized delivery of fragile therapeutics such as exosomes, whose rapid clearance severely limits efficacy. Addressing these bottlenecks requires biomaterials that can simultaneously rebuild three-dimensional structure and orchestrate a pro-regenerative microenvironment. Here, we present a morphology-adaptive injectable microporous annealed particle scaffold (MS) for localized and long-term release of human umbilical cord MSC-derived exosomes (hucMSC-Exo). GelMA/SFMA microspheres were fabricated via solvent-free microfluidic electrospraying combined with liquid nitrogen-assisted cryo-processing, preserving exosome bioactivity. This “disassemble-and-reassemble” approach enables minimally invasive injection of microspheres into irregular lesion cavities, followed by body temperature-induced annealing and in situ photocuring to form a mechanically stable, interconnected three-dimensional scaffold, which supports cell infiltration, nutrient diffusion, and sustained exosome release. In vivo, MS@Exo suppressed inflammation and scar formation, enhanced angiogenesis, and promoted neuronal survival, axonal regeneration, and myelination, ultimately achieving superior functional recovery over MS or Exo alone, demonstrating a clear synergistic effect. Transcriptomic profiling further revealed that MS@Exo reprogrammed gene expression by downregulating extracellular matrix- and collagen-related pathways while upregulating neuronal, synaptic, and ion channel signaling networks, thereby shifting the repair process from an inflammatory-proliferative phase toward a differentiation-remodeling phase. Collectively, MS@Exo acts as an intelligent multifunctional scaffold that couples advanced fabrication with exosome-based therapy to overcome fundamental limitations in SCI repair, offering a broadly applicable strategy for reconstructing irregular tissue defects in regenerative medicine.

Critical-sized bone defects (CSDs) represent a major clinical challenge due to their limited self-healing capacity. Conventional hydrogels incorporating crystalline hydroxyapatite (HA) often fail to recapitulate the hierarchical nanostructure of native bone, leading to suboptimal regeneration outcomes. To overcome this, we developed a biomimetic hydrogel by molecularly integrating amorphous calcium phosphate oligomers (CPO) into gelatin methacryloyl (GelMA), enabling a bone-like organic-inorganic hybrid network. This composite exhibits outstanding performance: an ultimate strength of 192 kPa at 18 wt% CPO confirms robust mechanical reinforcement, while 54% mass retention after 56 days underscores exceptional degradation resistance and dose-dependent bioactivity, evidenced by a two-fold upregulation of alkaline phosphatase (ALP) activity. The amorphous CPO facilitates biomimetic HA nucleation within the GelMA matrix, mimicking the natural mineralization process. In a rat calvarial CSD model, the hydrogel promoted 60% bone volume regeneration within 12 weeks, significantly outperforming conventional composites, through seamless host integration, vascularized trabecular bone formation, and a collagen-mineral hierarchy resembling native osteogenesis. This study establishes CPO as a transformative component that converts passive scaffolds into bioactive osteogenic microenvironments, offering a clinically viable strategy for complex bone regeneration.

Acute neuroinflammation drives secondary degeneration after spinal cord injury (SCI), yet the precise immune cell states and upstream regulatory circuits that initiate this response remain unresolved. Defining these early-state determinants at multi-omics resolution is essential for identifying mechanistically grounded therapeutic targets. We implemented an integrated multi-omics framework combining high-temporal-resolution single-cell RNA sequencing, bulk transcriptomics, histological validation, and systems-level network modeling across uninjured and early post-injury time points. Cell-cell communication analysis delineated intercellular signaling architecture within the acute lesion niche. Transcriptional regulatory network inference with in silico perturbation identified candidate master regulators. Network-based compound prioritization and target engagement validation were followed by functional testing in activated macrophages and a mouse SCI model. We resolved a temporally restricted S100a4+ macrophage state that rapidly emerged after injury, peaked at 1 day, and subsequently contracted. This state was defined by a coordinated transcriptional program integrating enhanced migratory capacity, amplified pro-inflammatory and pyroptotic signaling, and repression of homeostatic and reparative modules, constituting the dominant acute inflammatory signature at the tissue level. Systems-level analysis established a Cebpb-centered regulatory circuitry governing this state, thereby defining a C/EBPβ-S100a4+ macrophage axis as a principal driver of early neuroinflammation. Network topology positioned this axis as a densely connected and self-reinforcing hub within the injury microenvironment. Computational drug prioritization identified baicalein as a candidate regulator of C/EBPβ-dependent signaling. ChIP-qPCR and nuclear-cytoplasmic fractionation validated that baicalein effectively reduced the nuclear translocation of C/EBPβ and its binding to the S100a4 promoter. Experimental validation demonstrated that baicalein suppressed C/EBPβ expression, attenuated downstream inflammatory and pyroptotic pathways, and significantly improved functional recovery following SCI. This study delineates a C/EBPβ-S100a4+ macrophage axis that mechanistically structures the acute inflammatory landscape of SCI and represents a tractable therapeutic vulnerability. These findings advance a state-specific, network-informed framework for early immunomodulation in spinal cord injury.

Spinal cord injury (SCI) is a severe condition of the central nervous system that leads to significant functional impairments and imposes heavy socioeconomic burdens. Despite recent advancements, traditional treatments remain limited in terms of efficacy and are often associated with complications. Emerging therapies, including stem cell technologies and biomaterials, have shown promise in addressing the challenges associated with SCI recovery. Among these, hydrogels have gained considerable attention, owing to their excellent biocompatibility and adaptability, in addition to their ability to provide an optimal repair microenvironment. Importantly, hydrogels can serve as scaffolds for cell growth, drug delivery systems, or tools for modulating inflammatory responses and oxidative stress. This review comprehensively examines hydrogel-based therapies for the treatment of SCI, focusing on the mechanisms underlying secondary injury. The preparation of hydrogels and their application in promoting nerve regeneration, reducing scar formation, and mitigating oxidative damage are also presented. Furthermore, this review highlights the potential of hydrogels to facilitate macrophage polarization, increase the viability of stem cells, and enhance drug delivery precision. Despite these advancements, challenges remain, including optimization of material properties and achievement of targeted interventions. Future research should focus on developing smart hydrogels that adapt to the SCI microenvironment, investigate the integration of gene and physical therapies, and elucidate the molecular pathways involved in hydrogel-mediated regeneration. With continued innovation, hydrogels hold transformative potential in SCI treatment, being promising for improved functional recovery and patient outcomes.

The bidirectional interactions of spinal cord injury, multiple sclerosis, and amyotrophic lateral sclerosis with the gut operate through a distinct gut-spinal cord axis, rather than being fully explained by the conventional gut-brain axis. The spinal cord, with its unique anatomical and physiological features, serves as a central hub of communication. The gut and spinal cord communicate through various pathways, including the immune system and the autonomic and enteric nervous systems. This review summarizes existing clinical and basic research on the relationship between gut homeostasis and spinal cord diseases. First, we present findings from epidemiological studies showing that patients with spinal cord disorders often exhibit altered gut function, which may be influenced by antibiotic exposure and environmental factors. Second, we review the key physiological and anatomical structures of the gut-spinal cord axis, including the intestinal barrier, gut microbiota, and enteric nervous system, all of which are involved in maintaining gut health, as well as sensory neurons, motor neurons, and interneurons in spinal nerve regulation. Third, we describe the roles of the three axes (microbial, immune, and neural) in bidirectional regulation and their pathological mechanisms. Moreover, vicious cycles involving these axes can exacerbate spinal cord disorders. Fourth, we outline potential biomarkers in the gut-spinal cord axis, such as uridine, hypoxanthine, and 5-methoxytryptophan. Fifth, we propose several treatment strategies with potential clinical applications, including fecal microbiota transplantation and the use of probiotics and prebiotics. Finally, this review emphasizes the gut-spinal cord axis as a promising therapeutic target, highlighting the need for multi-omics integration, longitudinal cohort studies, and individualized interventions to resolve existing debates. Overall, the recognition of the gut-spinal cord axis provides a conceptual shift that extends beyond the gut-brain framework.

Objective: To explore the clinical characteristics and methods for syndrome differentiation prediction, as well as to construct a predictive model for Qi deficiency and blood stasis syndrome in patients with acute ischemic stroke (AIS). Methods: This study employed a retrospective case-control design to analyze patients with AIS who received inpatient treatment at the Neurology Department of The First Hospital of Hunan University of Chinese Medicine from January 1, 2013 to December 31, 2022. AIS patients meeting the diagnostic criteria for Qi deficiency and blood stasis syndrome were stratified into case group, while those without Qi deficiency and blood stasis syndrome were stratified into control group. The demographic characteristics (age and gender), clinical parameters [time from onset to admission, National Institutes of Health Stroke Scale (NIHSS) score, and blood pressure], past medical history, traditional Chinese medicine (TCM) diagnostic characteristics (tongue and pulse), neurological symptoms and signs, imaging findings [magnetic resonance imaging-diffusion weighted imaging (MRI-DWI)], and biochemical indicators of the two groups were collected and compared. The indicators with statistical difference (P < 0.05) in univariate analysis were included in multivariate logistic regression analysis to evaluate their predictive value for the diagnosis of Qi deficiency and blood stasis syndrome, and the predictive model was constructed by receiver operating characteristic (ROC) curve analysis. Results: The study included 1 035 AIS patients, with 404 cases in case group and 631 cases in control group. Compared with control group, patients in case group were significantly older, had extended onset-to-admission time, lower diastolic blood pressure, and lower NIHSS scores (P < 0.05). Case group showed lower incidence of hypertension history (P < 0.05). Regarding tongue and pulse characteristics, pale and dark tongue colors, white tongue coating, fine pulse, astringent pulse, and sinking pulse were more common in case group. Imaging examinations demonstrated higher proportions of centrum semiovale infarction, cerebral atrophy, and vertebral artery stenosis in case group (P < 0.05). Among biochemical indicators, case group showed higher proportions of elevated fasting blood glucose and glycated hemoglobin (HbA1c), while lower proportions of elevated white blood cell count, reduced hemoglobin, and reduced high-density lipoprotein cholesterol (HDL-C) (P < 0.05). Multivariate logistic regression analysis identified significant predictors for Qi deficiency and blood stasis syndrome including: fine pulse [odds ratio (OR) = 4.38], astringent pulse (OR = 3.67), superficial sensory abnormalities (OR = 1.86), centrum semiovale infarction (OR = 1.57), cerebral atrophy (OR = 1.55), vertebral artery stenosis (OR = 1.62), and elevated HbA1c (OR = 3.52). The ROC curve analysis of the comprehensive prediction model yielded an area under the curve (AUC) of 0.878 [95% confidence interval (CI) = 0.855 – 0.900]. Conclusion: This study finds out that Qi deficiency and blood stasis syndrome represents one of the primary types of AIS. Fine pulse, astringent pulse, superficial sensory abnormalities, centrum semiovale infarction, cerebral atrophy, vertebral artery stenosis, elevated blood glucose, elevated HbA1c, pale and dark tongue colors, and white tongue coating are key objective diagnostic indicators for the syndrome differentiation of AIS with Qi deficiency and blood stasis syndrome. Based on these indicators, a syndrome differentiation prediction model has been developed, offering a more objective basis for clinical diagnosis, and help to rapidly identify this syndrome in clinical practice and reduce misdiagnosis and missed diagnosis.

Spinal cord injury (SCI) is a matter of significant clinical concern, often treated through early surgical decompression along with methylprednisolone sodium succinate (MPSS). However, the side effects and the unsatisfactory focal concentration of MPSS have limited its further applications. To address this limitation, herein, a versatile drug delivery system of zeolitic imidazole framework-8 (ZIF-8) and gelatin methacryloyl microneedles (GelMA MNs) was developed for stable, transdural, and controlled sustained release of drugs in SCI. The microneedles were used to create tiny pores in the dura mater, allowing for the direct administration of drugs into the spinal cord. ZIF-8 provided a secondary extended release once they were separated from the microneedles. To attenuate the neuroinflammation, MPSS was selected. Such a combination of ZIF-8 and GelMA MNs was able to prolong the release period of MPSS to five days. The system showed transdural capacity, reduction of M1 polarization, and decrease in NLRP3-positive inflammasome and proinflammatory cytokines. In vivo studies indicated that this novel drug delivery strategy could constrict the inflammatory microenvironment, reduce glial scar formation, and promote neural regeneration. Thus, this versatile drug delivery system provides an up-and-coming alternative for stable, transdural, and controlled sustained release of drugs to those suffering from SCI.

Mesenchymal stem cells have made remarkable progress in recent years. Many studies have reported that human umbilical cord mesenchymal stem cells (hUC-MSCs) have no toxicity, but thromboembolism appeared in patients treated with hUC-MSCs. Therefore, people are still worried about the safety of clinical application. The study aims to determine the safety, potential toxic mechanism and biodistribution of hUC-MSCs. F344RG rats were given 5 or 50 million cells/kg of hUC-MSCs by single administration in compliance with Good Laboratory Practice standards. Standard toxicity was performed. RNA sequencing was then performed to explore the potential toxic mechanisms. In parallel, the biodistribution of hUC-MSCs was examined. The dose of 5 million cells/kg hUC-MSCs had no obvious toxicity on symptom, weight, food intake, hematology, serum biochemistry, urine biochemistry, cytokines, and histopathology. However, blood-tinged secretions in the urethral orifice and 20% mortality occurred at 50 million cells/kg. Disseminated intravascular coagulopathy (DIC) is the leading cause of death. hUC-MSCs significantly upregulated complement and coagulation cascade pathways gene expression, resulting in DIC. Besides, hUC-MSCs upregulated fibrinolytic system suppressor genes A2m, Serping1 and Serpinf2. hUC-MSCs survived in rats for less than 28 days, no hUC-MSC was detected in tissues outside the lungs. There was no toxicity in F344RG rats at 5 million cells/kg, but some toxicities were detected at 50 million cells/kg. hUC-MSCs significantly upregulated complement and coagulation cascade pathways, upregulated the expression of fibrinolytic system suppressor genes A2m, Serping1 and Serpinf2, to inhibit fibrinolytic system, caused DIC, which provided a new insight into the toxic mechanism of hUC-MSCs.

Purpose We present for the first time a novel entry point of pedicle screws (Short Rod Technique, SRT), which can avoid superior facet violation and has been verified as a safe screw placement method. The purpose of this study is to determine the clinical outcomes of SRT in posterior lumbar interbody fusion (PLIF) surgery. Methods We retrospectively analyzed the clinical outcomes of 89 patients who received SRT and 109 patients who received PLIF surgery with regular entry points of pedicle screws with a minimum of 2 years of follow-ups. Patients were divided into three groups according to the number of fusion segments, and the clinical outcomes of the three groups were compared. Results The length of the wound and the length of rods were significantly shorter in the each SRT group. Less intraoperative blood loss was observed in the SRT group in patients with a single segment and two segments fusions, but not in three segments fusions. Fewer degenerations of the upper adjacent segment were observed in the SRT group in patients with a single segment and three segments fusions. In addition, less postoperative wound pain related to PLIF surgery was observed in the SRT group in patients with two and three segments fusions. Conclusion SRT has been validated as an effective technique with good clinical outcomes, especially for reducing the occurrence of upper ASD in PLIF surgery with a single segment and three segments. The present study provides spinal surgeons with a novel method for performing PLIF surgery.

A new method of transdural delivering drugs to the spinal cord has been developed, involving the use of microneedles (MNs) and a β-cyclodextrin metal-organic framework (CD-MOF). This epidural microneedle array, dubbed MNs@CD-MOF@MPSS, can be utilized to deliver methylprednisolone sodium succinate (MPSS) to the site of spinal cord injury (SCI) in a controlled manner. MNs allows to generate micropores in the dura for direct drug delivery to the spinal cord, overcoming tissue barriers and targeting damaged regions. Additionally, the CD-MOF provides a secondary extended release after separating from the MNs. In in vitro study, inward MNs increased cellular absorption of MPSS and then reduced LPS-induced M1 polarization of microglia. And animal studies have shown that this method of drug delivery results in improved BMS scores and a reduction in M1 phenotype microphage and glial scar formation. Furthermore, the downregulation of the NLRP3-positive inflammasome and related pro-inflammatory cytokines was observed. In conclusion, this new drug platform has potential for clinical application in spinal cord diseases and is a valuable composite for minimally transdural controlled drug delivery. STATEMENT OF SIGNIFICANCE: This research presents a new epidural microneedle patch made up of microneedles (MNs) and a β-cyclodextrin metal-organic framework (CD-MOF). The epidural microneedle patch boasts high drug loading capacity, the ability to penetrate the dura, and controlled release. When loaded with methylprednisolone sodium succinate (MPSS), it effectively reduces inflammation and improves neurological function after spinal cord injury. Therefore, it is a novel and promising drug platform for the treatment of spinal cord diseases in a clinical setting.

Objective Improving accuracy and safety of pedicle screw placement is of great clinical importance. Electronic conductivity device (ECD) can be a promising technique with features of affordability, portability, and real‐time detection capabilities. This study aimed to validate the safety and effectiveness of a modified ECD. Methods The ECD underwent a modification where six lamps of various colors, and it was utilized in a prospectively multicenter randomized controlled clinical trial involving 96 patients across three hospitals from June 2018 to December 2018. The trial incorporated a self‐control randomization with an equal distribution of left or right side of vertebral pedicle among two groups: the free‐hand group and the ECD group. A total of 496 pedicle screws were inserted, with 248 inserted in each group. The primary outcomes focused on the accuracy of pedicle screw placement and the frequency of intraoperative X‐rays. Meanwhile, the secondary indicator measured the time required for pedicle screw placement. Results were presented as means ± SD. Paired samples t ‐test and χ 2 ‐test were used for comparison. Furthermore, an updated review was conducted, which included studies published from 2006 onwards. Results Baseline patient characteristics were recorded. The primary accuracy outcome revealed a 96.77% accuracy rate in the ECD group, compared to a 95.16% accuracy rate in the free‐hand group, with no significant differences noted. In contrast, ECD demonstrated a significant reduction in radiation exposure frequency when compared to the free‐hand group (1.11 ± 0.32 vs. 1.30 ± 0.53; p < 0.001), resulting in a 14.6% reduction. Moreover, ECD displayed a decrease of 30.38% in insertion time (70.88 ± 30.51 vs. 101.82 ± 54.00 s; p < 0.001). According to the results of the 21 studies, ECD has been utilized in various areas of the spine such as the atlas, thoracic and lumbar spine, as well as sacral 2‐alar‐iliac. The accuracy of ECD ranged from 85% to 100%. Conclusion The prospectively randomized trial and the review indicate that the use of ECD presents a secure and precise approach to the placement of pedicle screws, with the added benefit of reducing both procedure time and radiation exposure.

Calcium (Ca2+) signaling plays an important role in the regulation of many cellular functions. Ca2+-binding protein calmodulin (CaM) serves as a primary effector of calcium function. Ca2+/CaM binds to the death-associated protein kinase 1 (DAPK1) to regulate intracellular signaling pathways. However, the mechanism underlying the influence of Ca2+ on the conformational dynamics of the DAPK1−CaM interactions is still unclear. Here, we performed large-scale molecular dynamics (MD) simulations of the DAPK1−CaM complex in the Ca2+-bound and-unbound states to reveal the importance of Ca2+. MD simulations revealed that removal of Ca2+ increased the anti-correlated inter-domain motions between DAPK1 and CaM, which weakened the DAPK1−CaM interactions. Binding free energy calculations validated the decreased DAPK1−CaM interactions in the Ca2+-unbound state. Structural analysis further revealed that Ca2+ removal caused the significant conformational changes at the DAPK1−CaM interface, especially the helices α1, α2, α4, α6, and α7 from the CaM and the basic loop and the phosphate-binding loop from the DAPK1. These results may be useful to understand the biological role of Ca2+ in physiological processes.

Spinal cord injury (SCI) was a serious nerve injury, which involves complex genetic changes. This paper was intended to investigate the function and mechanism of differentially expressed genes in SCI. The three datasets GSE92657, GSE93561 and GSE189070 of SCI from GEO database were used to identify differentially expressed genes (DEGs). We identified the common DEGs in the three datasets GSE92657, GSE93561 and GSE189070 of SCI from GEO database. Next, a protein-protein interaction (PPI) network of DEGs was constructed. Subsequently, the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that DEGs were significantly enriched in immune response, inflammatory response. The expression level of immune-related genes (Arg1, Ccl12, Ccl2, Ifitm2, Ifitm3, and et al.) at different time points of SCI were analyzed in GSE189070 dataset. Next, differentially expressed miRNAs (DE-miRNAs) were identified in SCI compared with normal based on GSE158194 database. DE-miRNA and targeted immune-related genes were predicted by miRwalk, including miR-487b-5p targeted Ifitm3, miR-3072-5p targeted Ccl3, and et al. What’s more, the miR-487b was identified and verified to be down-regulated in Lipopolysaccharide (LPS)-induced BV-2 cell model. Further, the miR-487b inhibited cell inflammation and apoptosis in LPS-induced BV2 cell by targeted Ifitm3. For the first time, our results revealed that miR-487b may play an important regulatory role in SCI by targeted Ifitm3 and provide further evidence for SCI research.

青少年特发性脊柱侧凸(adolescent idiopathic scoliosis,AIS)是脊柱在三维结构上的畸形,好发于青少年女性.目前脊柱三维矫形手术仍是中重度AIS有效的治疗手段.AIS术后患者的生存质量评价越来越受到患者与脊柱外科医生的关注和重视,然而AIS好发于年轻女性且大多都处在生育年龄之前,目前对于AIS术后女性患者的妊娠相关问题研究较少,现有的研究结果中也存在一定的矛盾.本文总结该方面的研究,为广大医学工作者进一步的研究和探索提供一定的借鉴与参考.

Abstract Background Spinal cord injury (SCI) is an inflammatory condition, and excessive adenosine triphosphate (ATP) is released into the extracellular space, which can be catabolized into adenosine by CD73. Extracellular vesicles have been designed as nano drug carriers in many diseases. However, their impacts on delivery of CD73 after SCI are not yet known. We aimed to construct CD73 modified extracellular vesicles and explore the anti-inflammatory effects after SCI. Methods CD73 engineered extracellular vesicles (CD73+ hucMSC-EVs) were firstly established, which were derived from human umbilical cord mesenchymal stem cells (hucMSCs) transduced by lentiviral vectors to upregulate the expression of CD73. Effects of CD73+ hucMSC-EVs on hydrolyzing ATP into adenosine were detected. The polarization of M2/M1 was verified by immunofluorescence. Furthermore, A2aR and A2bR inhibitors and A2bR knockdown cells were used to investigate the activated adenosine receptor. Biomarkers of microglia and levels of cAMP/PKA were also detected. Repetitively in vivo study, morphology staining, flow cytometry, cytokine analysis, and ELISA assay, were also applied for verifications. Results CD73+ hucMSC-EVs reduced concentration of ATP and promoted the level of adenosine. In vitro experiments, CD73+ hucMSC-EVs increased macrophages/microglia M2:M1 polarization, activated adenosine 2b receptor (A2bR), and then promoted cAMP/PKA signaling pathway. In mice using model of thoracic spinal cord contusion injury, CD73+ hucMSC-EVs improved the functional recovery after SCI through decreasing the content of ATP in cerebrospinal fluid and improving the polarization from M1 to M2 phenotype. Thus, the cascaded pro-inflammatory cytokines were downregulated, such as TNF-α, IL-1β, and IL-6, while the anti-inflammatory cytokines were upregulated, such as IL-10 and IL-4. Conclusions CD73+ hucMSC-EVs ameliorated inflammation after spinal cord injury by reducing extracellular ATP, promoting A2bR/cAMP/PKA pathway and M2/M1 polarization. CD73+ hucMSC-EVs might be promising nano drugs for clinical application in SCI therapy. Graphical Abstract

ObjectiveTo explore the molecular regulatory mechanisms underlying fibroblast differentiation and dysfunction in the development of adolescent idiopathic scoliosis (AIS) in an effort to identify candidate therapeutic targets for AIS.MethodsThe GSE110359 dataset, obtained from the bone marrow stromal cells of 12 AIS patients and five healthy controls, was retrieved from the GEO database. The data were preprocessed and differentially expressed genes (DEGs) were identified. KEGG pathway and Gene Ontology (GO)‐Biological Process (BP) enrichment analyses were performed to identify the function of the DEGs. A protein–protein interaction (PPI) and a microRNA‐transcription factor (TF)‐target co‐regulatory network were constructed to identify hub genes in the development of AIS. In addition, hub DEGs were evaluated by quantitative PCR (qPCR) and immunohistochemical staining.ResultsA total of 188 DEGs including 100 up‐regulated and 88 down‐regulated genes were obtained. The up‐regulated DEGs were related to “p53 signaling pathway”, “FoxO signaling pathway”, and “cGMP‐PKG signaling pathway” terms, while the down‐regulated DEGs were significantly enriched in seven terms including “protein processing in endoplasmic reticulum”. The key up‐regulated genes, PRKG1, CCNG2, and KAT2B, and the key down‐regulated genes, MAP2K1 and DUSP6, were identified by the PPI and miRNA‐TF‐Target regulatory network analyses. mRNA expression patterns for PRKG1, DUSP6, and KAT2B were successfully verified by qPCR. In addition, PRKG1 protein levels were found to be elevated during the immunohistochemical analysis.ConclusionIncreased expression of PRKG1 in AIS patients might be an attractive therapeutic target for AIS. However, further gain or loss‐of‐function studies should be conducted.

Postmenopausal osteoporosis is caused by the deficiency of estrogen, which breaks bone homeostasis and induces levels of pro-inflammatory cytokines. Muscone is a potent anti-inflammatory agent and is used to treat bone fracture in traditional Chinese medicine. However, its anti-osteoclastogenic effects remain unclear. For in vitro study, morphology tests of osteoclastogenesis were firstly performed. And then, factors in RANK-induced NF-κB and MAPK pathways were examined by RT-PCR and Western blot, and the binding of TNF receptor-associated factor (TRAF)6 to RANK was inspected by coimmunoprecipitation and immunofluorescence staining. For in vivo experiments, C57BL/6 ovariectomized (OVX) mice were used for detection, including H&E staining, TRAP staining, and micro CT. As a result, muscone reduced OVX-induced bone loss in mice and osteoclast differentiation in vitro, by inhibiting TRAF6 binding to RANK, and then suppressed NF-κB and MAPK signaling pathways. The expression of the downstream biomarkers was finally inhibited, including NFATc1, CTR, TRAP, cathepsin K, and MMP-9. The inflammatory factors, TNF-a and IL-6, were also reduced by muscone. Taken together, muscone inhibited the binding of TRAF6 to RANK induced by RANKL, thus blocking NF-kB and MAPK pathways, and down-regulating related gene expression. Finally, muscone inhibited osteoclastogenesis and osteoclast function by blocking RANK-TRAF6 binding, as well as downstream signaling pathways in vitro. Muscone also reduced ovariectomy-induced bone loss in vivo.

BACKGROUND:Postmenopausal osteoporosis results from estrogen withdrawal and is characterized mainly by bone resorption. Shikonin is a bioactive constitute of Chinese traditional herb which plays a role in antimicrobial and antitumor activities. The study was designed to investigate the role of shikonin on postmenopausal osteoporosis and explore its underlying mechanisms.METHODS:Immunofluorescence staining was performed to evaluate the effects of shikonin on actin ring formation. The expression levels of the nuclear factor kappa-B (NF-κB) and mitogen-activated protein kinase (MAPK) pathway were determined by Western blot analysis. To determine whether shikonin influences the receptor activator of nuclear factor-κB ligand (RANKL)-induced association between receptor activator of NF-κB (RANK) and tumor necrosis factor receptor associated factor 6 (TRAF6), immunofluorescence staining and immunoprecipitation experiments were performed. During our validation model, histomorphometric examination and micro-computed tomography (CT) were conducted to assess the morphology of osteoporosis.RESULTS:Shikonin prevented bone loss by inhibiting osteoclastogenesis in vitro and improving bone loss in ovariectomized mice in vivo. At the molecular level, Western blot analysis indicated that shikonin inhibited the phosphorylation of inhibitor of NF-κB (IκB), P50, P65, extracellular regulated protein kinases (ERK), c-Jun N-terminal kinase (JNK), and P38. Interaction of TRAF6 and RANK was prevented, and downstream MAPK and NF-κB signaling pathways were downregulated.CONCLUSION:Osteoclastic bone resorption was reduced in the presence of shikonin in vitro and in vivo. Shikonin is a promising candidate for treatment of postmenopausal osteoporosis.

Postmenopausal osteoporosis (PMOP) has already become a major public health problem. However, there was no bibliometric analysis estimating the evolutionary process of PMOP research.PurposeThis study aimed to assess its scientific activity.MethodsPublications on PMOP were retrieved from Science Citation Index-Expanded (SCI-E) of the Web of Science (WoS) from 2008 to 2018. VOSviewer software was used for data mining and visualization. The quantity of papers, number of citations, citation frequency per year, and H-index were assessed and compared among different countries, institutes, and researchers.ResultsA total of 11,142 papers were included and were cited 184,416 times. The USA contributed the most papers (3162), the most citations (83,270 times), and the highest H-index (115). Canada ranked first for the number of papers per million populations and per trillion GDP. Seven of the top ten funding agencies were industries. The key words were stratified into four clusters: cluster 1 (basic research), cluster 2 (diagnosis), cluster 3 (treatment), and cluster 4 (others). Average appearing years (AAY) of key words in cluster 1 were the largest compared to those in clusters 2 and 3. For hotspots, miRNA showed a relatively latest AAY of 2017.63, followed by abaloparatide and adipokine.ConclusionWe concluded that the USA and Europe were the most productive regions on PMOP, with a high prevalence of articles supported by pharmaceutical companies. Key word focus gradually shifted from diagnosis to treatment and then to precision medicine orientated basic research. It is recommended to pay attention to potential research hotspots, such as miRNA, abaloparatide, and adipokine.