Phillygenin prevents osteoclast differentiation and bone loss by targeting RhoA.

Forsythia suspensa tea is a popular traditional Chinese medicine decoction for its healthy and therapeutic benefits. However, its effects in bone metabolism were not clear. In recent study, we uncovered anti-osteoclastogenesis property of Phillygenin (Phi), a compound abundant in Forsythia suspensa leaves, and aimed to investigate the effect and mechanism of Phi on bone metabolism in vivo and in vitro. Lipopolysaccharides-induced murine calvaria osteolysis and ovariectomy-induced bone loss animal models were used to identify the bone-protective effect of Phi in vivo and micro-CT, pQCT, and TRAP staining were applied. We used CCK8, TUNEL, BrdU, and TRAP staining to evaluate the efficacy of Phi on the proliferation and formation of OCs in primary mBMMs. RNA sequence, activity-based protein profiling, molecular docking, G-LISA, and WB were used to inspect the target and underlying mechanism of Phi's actions in mBMMs. We found Phi significantly inhibited bone resorption in vivo and inhibited mBMMs osteoclastogenesis in vitro. Ras homolog gene family member A (RhoA) was identified as the direct target of Phi. It counteracted the effects of RhoA activator and acted as a RhoA inhibitor. By targeting RhoA, Phi modulated Rho-associated coiled-coil containing protein kinase 1 (ROCK1) activity and regulated its downstream NF-κB/NFATc1/c-fos pathway. Furthermore, Phi depressed the disassembling of F-actin ring through cofilin and myosin1a. Our findings provided Phi as a potential option for treating bone loss diseases by targeting RhoA and highlighted the importance of F. suspensa as a preventive approach in bone disorders.

Advances in studies of ncRNAs in the pathophysiology and treatment of spinal disease.

Symptoms of spinal disease frequently accompany altered or damaged spine and vertebral structures resulting from endogenous and exogenous factors. Back braces, therapeutic agents, and surgery remain the main treatments for spinal diseases. However, the efficacy of currently available therapeutic agents is limited due to their side effects, whereas back braces and surgeries are less effective for certain patients. The significant effect of spinal disease on patients' morbidity and mortality emphasizes the necessity to develop novel and more effective therapeutic agents that mitigate the consequences of spinal disease. Accumulating research acknowledges that non-coding RNAs (ncRNAs), including miRNAs, lncRNAs, circRNAs, etc., are involved in the pathogenesis of spinal disease, their pronounced therapeutic potential and significant regulatory functions in spinal diseases. Hence, this review focuses on summarizing the latest advances in studies of ncRNAs in the progression and recovery of spinal diseases, as well as highlighting the collaboration of ncRNA networks in treating spinal disease.

Network Pharmacology to Identify the Pharmacological Mechanisms of a Traditional Chinese Medicine Derived from Trachelospermum jasminoides in Patients with Rheumatoid Arthritis.

BACKGROUND This study used a network pharmacology approach to identify the pharmacological mechanisms of a traditional Chinese medicine derived from Trachelospermum jasminoides (Lindl.) Lem. in patients with rheumatoid arthritis (RA). MATERIAL AND METHODS Known compounds of T. jasminoides were obtained from the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database, the Shanghai Institute of Organic Chemistry of Chinese Academy of Science, Chemistry (CASC) database, and a literature search. Putative targets of identified compounds were predicted by SwissTargetPrediction. RA-related targets were achieved from the Therapeutic Target database, Drugbank database, Pharmacogenomics Knowledgebase, and Online Mendelian Inheritance in Man database. The protein-protein interaction (PPI) network was built by STRING. CluGO was utilized for Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) enrichment analysis. RESULTS A total of 354 potential targets were predicted for the 17 bioactive compounds in T. jasminoides; 69 of these targets overlapped with RA-related targets. A PPI network was composed and 2 clusters of 59 and 42 nodes each were excavated. GO and KEGG enrichment analysis of the overlapping targets and the 2 clusters was mainly grouped into immunity, inflammation, estrogen, anxiety, and depression processes. CONCLUSIONS Our study illustrated that T. jasminoides alleviates RA through the interleukin-17 signaling pathway, the tumor necrosis factor signaling pathway, and other immune and inflammatory-related processes. It also may exert effects in regulating cell differentiation and potentially has anti-anxiety, anti-depression, and estrogen-like effects.

Lightweight Splint Design for Individualized Treatment of Distal Radius Fracture.

A systematic design approach is proposed for medical splints for individualized treatment of the distal radius fracture. An initial split structural model is first constructed by 3D scanning of an injured limb. Based on the biomechanical theory and clinical experiences, the topology optimization method is applied to design the splint structure. The optimized lightweight splint is realized by additive manufacturing using polylactic acid. Compared to the traditional designs for the distal radius fracture, the optimized design by the proposed approach exhibits a weight reduction of more than 40%. Besides, the mechanical properties of the splint meet the requirements of medical treatment according to the simulation results. Numerical examples are provided to demonstrate the applicability of the approach.

Homochiral silver-based coordination polymers exhibiting temperature-dependent photoluminescence behavior.

Homochiral coordination polymers based on zigzag or helical polymers are assembled from a semiflexible fluorescent bipyridyl ligand and linear coordinated Ag(I) ions; they exhibit unusual temperature-dependent photoluminescence behavior, including multistep changes in energy and intensity upon cooling.

Chiral metal-organic frameworks bearing free carboxylic acids for organocatalyst encapsulation.

Two chiral carboxylic acid functionalized micro- and mesoporous metal-organic frameworks (MOFs) are constructed by the stepwise assembly of triple-stranded heptametallic helicates with six carboxylic acid groups. The mesoporous MOF with permanent porosity functions as a host for encapsulation of an enantiopure organic amine catalyst by combining carboxylic acids and chiral amines in situ through acid-base interactions. The organocatalyst-loaded framework is shown to be an efficient and recyclable heterogeneous catalyst for the asymmetric direct aldol reactions with significantly enhanced stereoselectivity in relative to the homogeneous organocatalyst.

Trachelogenin alleviates osteoarthritis by inhibiting osteoclastogenesis and enhancing chondrocyte survival.

BACKGROUND: Osteoarthritis (OA) is a prevalent global health concern associated with the loss of articular cartilage and subchondral bone. The lack of disease-modifying drugs for OA necessitates the exploration of novel therapeutic options. Our previous study has demonstrated that traditional Chinese medical herb Trachelospermum jasminoides (Lindl.) Lem. extract suppressed osteoclastogenesis and identified trachelogenin (TCG) as a representative compound. Here, we delved into TCG's potential to alleviate OA. METHODS: We initially validated the in vivo efficacy of TCG in alleviating OA using a rat OA model. Subsequently, we isolated primary bone marrow-derived macrophages in vitro to investigate TCG's impact on osteoclastogenesis. We further employed a small molecule pull-down assay to verify TCG's binding target within osteoclasts. Finally, we isolated primary mouse chondrocytes in vitro to study TCG's regulatory effects and mechanisms on chondrocyte survival. RESULTS: TCG preserved subchondral bone integrity and protected articular cartilage in a rat OA model. Subsequently, in vitro experiments unveiled TCG's capability to inhibit osteoclastogenesis and function through binding to Ras association proximate 1 (Rap1) and inhibiting its activation. Further study demonstrated that TCG inhibited Rap1/integrin αvß3/c-Src/Pyk2 signaling cascade, and consequently led to failed F-actin ring formation. Besides, TCG promoted the proliferation of mouse primary chondrocytes while suppressing apoptosis in vitro. This is attributed to TCG's ability to upregulate HIF1α, thereby promoting glycolysis. CONCLUSION: TCG exerted inhibitory effects on osteoclastogenesis through binding to Rap1 and inhibiting Rap1 activation, consequently preventing subchondral bone loss. Moreover, TCG enhanced chondrocyte survival by upregulating HIF1α and promoting glycolysis. These dual mechanisms collectively provide a novel approach to prevented against cartilage degradation.

Topology-Optimized Splints vs Casts for Distal Radius Fractures: A Randomized Clinical Trial.

Importance: To date, there is currently no evidence-based medical support for the efficacy of topology-optimized splints in treating distal radius fractures. Objective: To assess the clinical efficacy and complication rates of topology-optimized splints in the treatment of distal radius fractures after closed manual reduction. Design, Setting, and Participants: This 12-week, multicenter, open-label, analyst-blinded randomized clinical trial (comprising a 6-week intervention followed by a 6-week observational phase) was carried out from December 3, 2021, to March 10, 2023, among 110 participants with distal radius fractures. Statistical analysis was performed on an intention-to-treat basis between June 3 and 30, 2023. Intervention: Participants were randomly assigned to 2 groups: the intervention group received topology-optimized splint immobilization and the control group received cast immobilization after closed manual reduction for 6weeks. After this period, immobilization was removed, and wrist rehabilitation activities commenced. Main Outcomes and Measures: The primary outcome was the Gartland-Werley (G-W) wrist score at 6 weeks (where higher scores indicate more severe wrist dysfunction). Secondary outcomes encompassed radiographic parameters, visual analog scale scores, swelling degree grade, complication rates, and 3 dimensions of G-W wrist scores. Results: A total of 110 patients (mean [SD] age, 64.1 [12.7] years; 89 women [81%]) enrolled in the clinical trial, and complete outcome measurements were obtained for 101 patients (92%). Median G-W scores at 6 weeks were 15 (IQR, 13-18) for the splint group and 17 (IQR, 13-18) for the cast group (mean difference, -2.0 [95% CI, -3.4 to -0.6]; P = .03), indicating a statistically significant advantage for the splint group. At 12 weeks, no clinically significant differences in G-W scores between the 2 groups were observed. Complication rates, including shoulder-elbow pain and dysfunction and skin irritation, were less common in the splint group (shoulder-elbow pain and dysfunction: risk ratio, 0.28 [95% CI, 0.08-0.93]; P = .03; skin irritation: risk ratio, 0.30 [95% CI, 0.10-0.89]; P = .02). Conclusions and Relevance: Findings of this randomized clinical trial suggest that patients with distal radius fractures that were managed with topology-optimized splints had better wrist functional outcomes and fewer complications at 6 weeks compared with those who received casting, with no difference at week 12. Therefore, topology-optimized splints with improved performance have the potential to be an advisable approach in the management of distal radius fractures. Trial Registration: Chinese Clinical Trial Registry: ChiCTR2000036480.

MAPK8 and CAPN1 as potential biomarkers of intervertebral disc degeneration overlapping immune infiltration, autophagy, and ceRNA.

Background: Intervertebral disc degeneration (IDD) is one of the most common health problems in the elderly and a major causative factor in low back pain (LBP). An increasing number of studies have shown that IDD is closely associated with autophagy and immune dysregulation. Therefore, the aim of this study was to identify autophagy-related biomarkers and gene regulatory networks in IDD and potential therapeutic targets. Methods: We obtained the gene expression profiles of IDD by downloading the datasets GSE176205 and GSE167931 from the Gene Expression Omnibus (GEO) public database. Subsequently, differentially expressed genes (DEGs) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, gene ontology (GO), and gene set enrichment analysis (GSEA) were performed to explore the biological functions of DEGs. Differentially expressed autophagy-related genes (DE-ARGs) were then crossed with the autophagy gene database. The hub genes were screened using the DE-ARGs protein-protein interaction (PPI) network. The correlation between the hub genes and immune infiltration and the construction of the gene regulatory network of the hub genes were confirmed. Finally, quantitative PCR (qPCR) was used to validate the correlation of hub genes in a rat IDD model. Results: We obtained 636 DEGs enriched in the autophagy pathway. Our analysis revealed 30 DE-ARGs, of which six hub genes (MAPK8, CTSB, PRKCD, SNCA, CAPN1, and EGFR) were identified using the MCODE plugin. Immune cell infiltration analysis revealed that there was an increased proportion of CD8+ T cells and M0 macrophages in IDD, whereas CD4+ memory T cells, neutrophils, resting dendritic cells, follicular helper T cells, and monocytes were much less abundant. Subsequently, the competitive endogenous RNA (ceRNA) network was constructed using 15 long non-coding RNAs (lncRNAs) and 21 microRNAs (miRNAs). In quantitative PCR (qPCR) validation, two hub genes, MAPK8 and CAPN1, were shown to be consistent with the bioinformatic analysis results. Conclusion: Our study identified MAPK8 and CAPN1 as key biomarkers of IDD. These key hub genes may be potential therapeutic targets for IDD.

Psoralen suppresses the phosphorylation of amyloid precursor protein (APP) to inhibit myelosuppression.

This study aims to explore the effect of Psoralen on myelosuppression, and investigating the mechanism involved in. The mesenchymal stem cells (MSCs) were treated with CTX to construct cell model of myelosuppression, and then with APP knockdown or overexpression transfection. Cell proliferation, cell apoptosis, bone growth factors, and hematopoietic growth factors were identified. The animal model of myelosuppression syndrome was established by intraperitoneal injection of cyclophosphamide (CTX) into C57BL/6 mice, and then with APP knockdown transfection. The effect of Psoralen on myelosuppression mice with APP knockdown was explored, including observin the number of hematopoietic stem cells and bone marrow MSCs, detecting the degree of osteoporosis and the number of osteoclasts. The expression of phosphorylation-amyloid precursor protein (p-APP), bone growth factors, and hematopoietic growth factors were also examined. We found that CTX treatment inhibited cell proliferation, induced cell apoptosis, promoted p-APP/APP, and inhibited the expression of aph-1 homolog A (APH-1α), presenilin enhancer-2 (PEN-2), the receptor of advanced glycation endproducts (RAGE). Psoralen pretreatment effectively promoted cell proliferation, suppressed cell apoptosis, inhibited p-APP/APP and stimulated the expression of APH-1α, PEN-2, RAGE compared with CTX treatment. After APP knockdown, cell proliferation was inhibited, and cell apoptosis was increased. The release of bone growth factors and hematopoietic growth factors was decreased. Psoralen pretreatment could reverse the effect of APP knockdown on MSCs and myelosuppression mice. In conclusion, Psoralen treatment inhibited cell apoptosis and regulated bone growth factors and hematopoietic growth factors in myelosuppression syndrome by suppressing the phosphorylation of APP.

Transporting Hydrogel via Chinese Acupuncture Needles for Lesion Positioning Therapy.

Lesion positioning therapy optimizes medical treatment by directly targeting lesions. However, strong physical barriers greatly hinder its wide use. Here, the Chinese acupuncture needles (CA-needles) with a screw-thread structure at the tip (ST-needle) and the hydrogel with the function of adhesive metal and loaded drug sustained-release structure are designed, through the minimally invasive and precise positioning of lesions by ST-needles, the dry-wet conversion of hydrogel with absorbing fluids and swelling, and the rotation back of ST-needles, the hydrogel is precisely positioned in the subchondral bone with physical barrier to achieve precise positioning therapy for lesions. In vitro experiments show that the ST-needle penetrates the physical barrier of cartilage and enters the subchondral bone. Simultaneously, the hydrogel transfer efficiency of the ST-needle (73.25%) is significantly higher than that of the CA-needle (29.92%) due to the protective effect of the screw-thread structure. In vivo experiments demonstrate that precise positioning in subchondral bone in osteoarthritis rats with ST-needles effectively inhibits abnormal subchondral bone remodeling, alleviating the degeneration and degradation of cartilage. Therefore, ST-needles achieve lesion positioning therapy through minimally invasive penetration of physical barriers, precisely positioning within lesions, and delivering hydrogel to release drugs.

Cynarin suppresses gouty arthritis induced by monosodium urate crystals.

The study is aimed to determine the effects of cynarin (Cyn) on mice with gouty arthritis (GA) induced by monosodium urate (MSU). We measured swelling in the hind paws of mice in vivo using Vernier calipers and ultrasound. The liver, kidney, and hind paws were stained with hematoxylin-eosin, and M1 type macrophages were detected in the hind paws using anti-F4/80 and anti-iNOS antibodies. The mRNA expression of inflammatory factors in bone marrow-derived macrophages (BMDMs) and in the hind paws was detected via quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Nucleotide-binding oligomerization domain (NOD)-like receptor (NLR) family pyrin domain containing 3 (NLRP3) inflammasomes and the nuclear factor kappa B (NF-κB) and mitogen-activated protein kinase (MAPK) pathways were analyzed via western blotting. Cyn was detected in vitro using Cell Counting Kit-8 (CCK-8). Cyn treatment reduced hind paw swelling and M1 macrophage infiltration, suppressed the mRNA expression of inflammatory factors, and inhibited NLRP3 inflammasome activation in vivo, in addition to inhibiting the phosphorylation of IKKa/ß, p65, and c-Jun NH 2-terminal kinase (JNK). Furthermore, Cyn exerted anti-inflammatory and anti-swelling effects in mice with GA by regulating the NF-κB and JNK pathways and NLRP3 inflammasomes.

α-Asarone Attenuates Osteoclastogenesis and Prevents Against Oestrogen-Deficiency Induced Osteoporosis.

Osteoporosis (OP) is defined as low bone mineral density which features over activated osteoclasts (OCs) and bone resorption. Targeting excessive OCs activity is thought to be an effective therapeutic approach for OP treatment. α-asarone (ASA), a compound from the traditional Chinese medicinal herb Acorus tatarinowii, has been widely used as a therapeutic agent against several diseases such as epilepsy, cough, bronchitis and asthma for many years. Recently, it was reported that ASA-derived lignins which were purified from Acorus tatarinowii root tissues effectively suppressed both RANKL-induced osteoclastogenesis and bone resorption. Besides, a classic Chinese formulation Bajitianwan (BJTW) which consisted of root and rhizome of Acorus tatarinowii Schott also showed positive effects on age-related bone loss. In the present study, we aimed to study the effects of ASA on osteoclastogenesis in vitro and in vivo. As illustrated by TRAP staining, ASA was capable of inhibiting RANKL-induced osteoclastogenesis in a dose-dependent manner, not only at an early-stage, but also in the late-stage. Besides, it also effectively suppressed bone resorption of mature OCs in a pit resorption assay. The formation of F-actin ring during osteoclastogenesis, which was important in OCs bone-resorption, was impaired as well. Subsequent mechanism experiments exposed that ASA inhibited osteoclastogenesis related genes in a time-dependent manner through AKT, p38 and NF-κB, followed by NFATc1/c-fos signaling pathway. Notably, our in vivo study uncovered that ASA was capable of improving the bone microstructure in oestrogen-deficiency induced OP models. Thus, our current work highlighted the important role of an old drug ASA in bone metabolism especially in OCs differentiation. ASA may find its potential as a lead compound to treat excessive OCs activity-induced bone loss diseases and more structure optimization is further needed.

The extract of Trachelospermum jasminoides (Lindl.) Lem. vines inhibits osteoclast differentiation through the NF-κB, MAPK and AKT signaling pathways.

Osteoclasts are the only cells in the body with a bone-resorption function. The identification of anti-osteoclastogenic agents is important in managing bone loss diseases. The dried vines of Trachelospermum jasminoides (Lindl.) Lem. have been used as a herbal medicine to treat musculoskeletal soreness in East Asia for hundreds of years. In the present study, we focused on the effect of Trachelospermum jasminoides (Lindl.) Lem. extract (TJE) on osteoclast differentiation. As indicated by tartrate-resistant acid phosphatase (TRAP) staining, TJE inhibited osteoclastogenesis induced by receptor activator of nuclear factor-κB ligand from bone marrow-derived monocytes/macrophages without showing any cytotoxicity. In addition, TJE effectively suppressed F-actin ring formation and the bone-resorption function of osteoclasts. The subsequent studies such as network pharmacology and molecular investigation, revealed that TJE inhibited osteoclastogenesis-related genes in a dose- and time-dependent manner through NF-κB, MAPK and AKT-mediated mechanism followed by the nuclear factor of activated T cells, cytoplasmic 1 (NFATc1)/c-Fos pathway. Our study could potentially explain the underlying molecular pharmacology of TJE in osteoclast-related diseases. What's more, it suggested that network pharmacology could help the modernization of traditional Chinese medicine.

Selective binding and removal of organic molecules in a flexible polymeric material with stretchable metallosalen chains.

A dynamic porous coordination network with a hydrophobic channel was assembled from stretchable 1D metallosalen polymer, which has the ability to recognize and, particularly, separate aromatic hydrocarbons from aliphatic mixtures with high selectivity.

Diagnostic value of T1ρ and T2 mapping sequences of 3D fat-suppressed spoiled gradient (FS SPGR-3D) 3.0-T magnetic resonance imaging for osteoarthritis.

Three-dimensional fat-suppressed spoiled gradient magnetic resonance imaging can be used to observe cartilages with high resolution.To quantify and compare the T1ρ and T2 relaxation times of the knee articular cartilage between healthy asymptomatic adults and patients with osteoarthritis (OA).This was a retrospective study of 53 patients with symptomatic OA (6 males and 47 females; aged 57.6 ±â€Š10.0 years) and 26 healthy adults (11 males and 15 females; aged 31.7 ±â€Š12.2 years) from the Ruijin Hospital. T1ρ and T2 relaxation times of knee cartilage were quantified using sagittal multi-echo T1ρ and T2 mapping sequences (3.0-T scanner) and analyzed by receiver operating characteristic (ROC) curve.T1ρ and T2 relaxation times in the OA group were higher than in controls (both P < .01). The sensitivity, specificity, and critical value for differentiating normal from OA cartilage were respectively 92%, 85.6%, and 45.90 ms for T1ρ, and 93.6%, 93.3%, and 50.42 ms for T2. T2 mapping sequence showed a higher area under the ROC curve (AUC) than T1ρ (0.965 vs 0.927, P = .02). The AUC for differentiating normal from Noyes IIA cartilage was 0.922 for T1ρ (cut-off: 46.0; sensitivity: 87.7%; specificity: 89.7%) and 0.954 for T2 (cut-off: 49.5; sensitivity: 91.2%; specificity: 92.3%), with no significant difference between them (P = .08).Both T1ρ and T2 mapping sequences could be used to assess OA cartilage lesions, with T2 mapping sequence demonstrating significant sensitivity for cartilage degeneration. These 2 sequences could also identify early-stage OA cartilage.

A Novel Rhein Derivative Modulates Bone Formation and Resorption and Ameliorates Estrogen-Dependent Bone Loss.

Osteoporosis, an osteolytic disease that affects millions of people worldwide, features a bone remodeling imbalance between bone resorption by osteoclasts and bone formation by osteoblasts. Identifying dual target-directed agents that inhibit excessive bone resorption and increase bone formation is considered an efficient strategy for developing new osteoporosis treatments. Rhein, a natural anthraquinone, can be isolated from various Asian herbal medicines. Rhein and its derivatives have been reported to have various beneficial pharmacological effects, especially their bone-targeting ability and anti-osteoclastogenesis activity. Moreover, hydrogen sulfide (H2 S) was reported to prevent ovariectomy- (OVX-) induced bone loss by enhancing bone formation, and sulfur replacement therapy has been considered a novel and plausible therapeutic option. Based on this information, we synthesized a rhein-derived thioamide (RT) and investigated its effects on bone resorption and bone formation in vitro and in vivo. It has been found that the RT-inhibited receptor activator of the nuclear factor-κB (NF-κB) ligand- (RANKL-) induced osteoclastogenesis and bone resorption in a dose-dependent manner. The expression of osteoclast marker genes was also suppressed by RT treatment. Furthermore, exploration of signal transduction pathways indicated that RT markedly blocked RANKL-induced osteoclastogenesis by attenuating MAPK pathways. However, RT treatment in an osteoblastic cell line, MC3TE-E1, indicated that RT led to an increase in the deposition of minerals and the expression of osteoblast marker genes, as demonstrated by Alizarin Red staining and alkaline phosphatase activity. Importantly, an OVX mouse model showed that RT could attenuate the bone loss in estrogen deficiency-induced osteoporosis in vivo with a smart H2 S-releasing property and that there was a considerable improvement in the biomechanical properties of bone. Accordingly, our current work highlights the dual regulation of bone remodeling by the rhein-derived molecule RT. This may be a highly promising approach for a new type of anti-osteoporosis agent. © 2018 American Society for Bone and Mineral Research.

MicroRNA-9 controls apoptosis of neurons by targeting monocyte chemotactic protein-induced protein 1 expression in rat acute spinal cord injury model.

OBJECTIVE: For the purpose of an early identification of intervention targets for acute spinal cord injury (ASCI), we investigated the changes in expression levels of microRNA-9 (miR-9) and MCPIP1 in rat ASCI model. METHOD: A total of 108 healthy rats were randomly divided into non-ASCI group (n=18) and five ASCI groups, 6h, 12h, 24h, 3 days and 7 days, representing the experimental time points following ASCI (n=18 per group). Hematoxylin and eosin (HE) staining was used to assess the ASCI damage, and quantitative real-time PCR (qRT-PCR) and in situ hybridization (ISH) were employed for the detection of miR-9 and MCPIP1 mRNA expression. RESULTS: HE staining results showed normal neuronal morphology in the non-ASCI group, but spinal cord tissue at 6h after ASCI showed developing neuronal necrosis. Acute inflammatory response was evident at 12h and 24h, with immune cells infiltrating into the gray matter. Vascular permeability increased and the nerve cells in gray-white matter exhibited extensive damage and necrosis at 24h and 7 days after ASCI. MiR-9 expression in ASCI tissue was significantly lower than that in normal spinal cord tissue. Statistical analysis showed a significant decrease in miR-9 expression in all the ASCI groups, compared to the non-ASCI group. Results from real-time PCR analysis revealed that MCPIP1 expression in all the ASCI groups was significantly higher than the non-ASCI group, and MCPIP1 expressions gradually increased with times at 6h-24h after ASCI. ISH revealed the following results after ASCI (1) miR-9 and MCPIP1 mRNA expression mainly distributed in ventral horn motor neurons, (2) miR-9 expression was high at 7 day after ASCI and (3) in the non-ASCI group, MCPIP1 expression was high at 6h, 12h, 24h and 3 days. CONCLUSION: MCPIP1 is significantly up-regulated after ASCI. The negative relationship between MCPIP1 and miR-9 suggest that MCPIP1 mRNA could be a target of miR-9 during ASCI. Thus, miR-9 is a marker for apoptosis in neurons, and an excellent therapeutic target for ASCI patients.