Metformin promotes microglial cells to facilitate myelin debris clearance and accelerate nerve repairment after spinal cord injury.

Spinal cord injury (SCI) is one kind of severe trauma for central nervous system. Myelin debris clearance and axon regeneration are essential for nerve regeneration after SCI. Metformin, a glucose-lowering drug, has been demonstrated to promote the locomotor functional recovery after SCI. In this study, we investigated the role and molecular mechanism of metformin on myelin preservation in a rat SCI model. SCI was induced in rats by compression at T9 level using a vascular clip. We showed that administration of metformin (50 mg·kg-1·d-1, ip) for 28 days significantly improved locomotor function in SCI rats. Metformin also ameliorated SCI-induced neuronal apoptosis and promoted axon regeneration in the spinal cord. Using co-immunofluorescence of IBa-1 and MBP, and luxol fasting blue (LFB) staining, we demonstrated that metformin promoted the transformation of M1 to M2 phenotype polarization of microglial cells, then greatly facilitated myelin debris clearance and protected the myelin in SCI rats. Furthermore, metformin ameliorated SCI-induced blockade of autophagic flux in the spinal cord, and enhanced the fusion of autophagosome and lysosome by inhibiting the AMPK-mTOR signaling pathway. Moreover, metformin significantly attenuated inflammatory responses in the spinal cord. In LPS-treated BV2 cells, pretreatment with metformin (2 mM) significantly enhanced autophagy level, suppressed inflammation and cell apoptosis. The protective effects were blocked in the presence of an autophagy inhibitor 3-methyladenine (3-MA, 5 mM), suggesting that the effect of metformin on autophagy in microglial cells is essential for the myelin preservation during nerve recovery. This study reveals a novel therapeutic effect of metformin in SCI recovery by regulating the activation of microglial cells and enhancing its autophagy level.

Retracted: Therapeutic Effects of Resveratrol Liposome on Muscle Injury in Rats.

This publication has been retracted by the Editor due to concerns regarding the originality of the figure images.Reference:Yongzeng Feng, Zili He, Cong Mao, Xiaolong Shui, Leyi Cai. Therapeutic Effects of Resveratrol Liposome on Muscle Injury in Rats. Med Sci Monit, 2019; 25:2377-2385. DOI: 10.12659/MSM.913409.

A Cascade Attention Based Facial Expression Recognition Network by Fusing Multi-Scale Spatio-Temporal Features.

The performance of a facial expression recognition network degrades obviously under situations of uneven illumination or partial occluded face as it is quite difficult to pinpoint the attention hotspots on the dynamically changing regions (e.g., eyes, nose, and mouth) as precisely as possible. To address the above issue, by a hybrid of the attention mechanism and pyramid feature, this paper proposes a cascade attention-based facial expression recognition network on the basis of a combination of (i) local spatial feature, (ii) multi-scale-stereoscopic spatial context feature (extracted from the 3-scale pyramid feature), and (iii) temporal feature. Experiments on the CK+, Oulu-CASIA, and RAF-DB datasets obtained recognition accuracy rates of 99.23%, 89.29%, and 86.80%, respectively. It demonstrates that the proposed method outperforms the state-of-the-art methods in both the experimental and natural environment.

EEG Emotion Classification Network Based on Attention Fusion of Multi-Channel Band Features.

Understanding learners' emotions can help optimize instruction sand further conduct effective learning interventions. Most existing studies on student emotion recognition are based on multiple manifestations of external behavior, which do not fully use physiological signals. In this context, on the one hand, a learning emotion EEG dataset (LE-EEG) is constructed, which captures physiological signals reflecting the emotions of boredom, neutrality, and engagement during learning; on the other hand, an EEG emotion classification network based on attention fusion (ECN-AF) is proposed. To be specific, on the basis of key frequency bands and channels selection, multi-channel band features are first extracted (using a multi-channel backbone network) and then fused (using attention units). In order to verify the performance, the proposed model is tested on an open-access dataset SEED (N = 15) and the self-collected dataset LE-EEG (N = 45), respectively. The experimental results using five-fold cross validation show the following: (i) on the SEED dataset, the highest accuracy of 96.45% is achieved by the proposed model, demonstrating a slight increase of 1.37% compared to the baseline models; and (ii) on the LE-EEG dataset, the highest accuracy of 95.87% is achieved, demonstrating a 21.49% increase compared to the baseline models.

An Improved Tiered Head Pose Estimation Network with Self-Adjust Loss Function.

As an important task in computer vision, head pose estimation has been widely applied in both academia and industry. However, there remains two challenges in the field of head pose estimation: (1) even given the same task (e.g., tiredness detection), the existing algorithms usually consider the estimation of the three angles (i.e., roll, yaw, and pitch) as separate facets, which disregard their interplay as well as differences and thus share the same parameters for all layers; and (2) the discontinuity in angle estimation definitely reduces the accuracy. To solve these two problems, a THESL-Net (tiered head pose estimation with self-adjust loss network) model is proposed in this study. Specifically, first, an idea of stepped estimation using distinct network layers is proposed, gaining a greater freedom during angle estimation. Furthermore, the reasons for the discontinuity in angle estimation are revealed, including not only labeling the dataset with quaternions or Euler angles, but also the loss function that simply adds the classification and regression losses. Subsequently, a self-adjustment constraint on the loss function is applied, making the angle estimation more consistent. Finally, to examine the influence of different angle ranges on the proposed model, experiments are conducted on three popular public benchmark datasets, BIWI, AFLW2000, and UPNA, demonstrating that the proposed model outperforms the state-of-the-art approaches.

Paeoniflorin promotes angiogenesis and tissue regeneration in a full-thickness cutaneous wound model through the PI3K/AKT pathway.

The treatment of wounds remains a clinical challenge because of poor angiogenesis under the wound bed, and increasingly, the patients' need for functional and aesthetically pleasing scars. For the wound healing process, new blood vessels which can deliver nutrients and oxygen to the wound area are necessary. In this study, we investigated the pro-angiogenesis ability and mechanism in wound healing of paeoniflorin (PF), which is a traditional Chinese medicine. In our in vitro results, the ability for proliferation, migration and in vitro angiogenesis in human umbilical vein endothelial cells was promoted by coculturing with PF (1.25-5 µM). Meanwhile, molecular docking studies revealed that PF has excellent binding abilities to phosphatidylinositol-3-kinase (PI3K) and protein kinase B (AKT), and consistent with our western blot results, that PF suppressed PI3K and AKT phosphorylation. Furthermore, to investigate the healing effect of PF in vivo, we constructed a full-thickness cutaneous wound model in rats. PF stimulated the cellular proliferation status, collagen matrix deposition and remodeling processes in vitro and new blood vessel formation at the wound bed resulting in efficient wound healing after intragastric administration of 10 mg·kg-1 ·day-1 in vivo. Overall, PF performed the pro-angiogenetic effect in vitro and accelerating wound healing in vivo. In summary, the capacity for angiogenesis in endothelial cells could be enhanced by PF treatment via the PI3K/AKT pathway in vitro and could accelerate the wound healing process in vivo through collagen deposition and angiogenesis in regenerated tissue. This study provides evidence that application of PF represents a novel therapeutic approach for the treatment of cutaneous wounds.

Chlorobenzene levels, component distribution, and ambient severity in wastewater from five textile dyeing wastewater treatment plants.

Chlorobenzenes (CBs) present in synthetic dyes are discharged into natural waters during the treatment of textile dyeing wastewater, which may have adverse effects on human and environment. In this study, the existence and removal of 12 CBs in different units of five treatment plants were examined. The ecological risk of CBs in textile dyeing wastewater was assessed by ambient severity (AS) and risk quotients (RQs). The results showed that trichlorobenzene, tetrachlorobenzene, pentachlorobenzene and hexachlorobenzene were ubiquitous in textile dyeing wastewater, and their distribution was similar. In one of the plants, the content of hexachlorobenzene was found to be as high as 9.277 µg/L in the raw water, which was an oil-water mixture. In other plants, there was no significant difference in the content and composition of CBs among influent and effluent suggesting that the conventional wastewater treatment plants cannot improve the existence of them. Monochlorobenzene and dichlorobenzene were not detected, which may have been related to strong volatility, biochemical properties, and weak instrument sensitivity. In the treatment process and effluent, trichlorobenzene is the main pollutant and accounted for 39.51% of all CB. CB removal was found only in the anaerobic system, while the aerobic system did not have the corresponding removal effect on CB and total organic carbon. According to ecological risk assessment, CBs in effluent has not been found the significant potential harm to human health (AS < 1), but posed moderate ecological risk to aquatic ecosystem (RQs > 0.1).

HLMethy: a machine learning-based model to identify the hidden labels of m6A candidates.

KEY MESSAGE: We developed a machine learning-based model to identify the hidden labels of m6A candidates from noisy m6A-seq data. Peak-calling approaches, such as MeRIP-seq or m6A-seq, are commonly used to map m6A modifications. However, these technologies can only map m6A sites with 100-200 nt resolution and cannot reveal the precise location or the number of modified residues in a transcript. To address this challenge, we developed a novel machine learning-based approach, named HLMethy, to assign labels to m6A candidates from noisy m6A-seq data. The multiple instance learning framework was adopted and two different training strategies were used to generate the classification model. To test the performance of our model, the m6A sites with single-base resolution were used and our model achieved comparable performance against existing instance-level predictors, which suggest that our model has the potential to improve the data quality of m6A-seq at reduced costs. What's more, our generic framework can be extended to other newly found modifications that are found by peak-calling approaches. The source code of HLMethy is available at https://github.com/liuze-nwafu/HLMethy.

Therapeutic Effects of Resveratrol Liposome on Muscle Injury in Rats.

BACKGROUND In this study we prepared liposome microbubbles loading resveratrol (LMLR) and evaluated its therapeutic effect on injury of gastrocnemius muscle in rats. MATERIAL AND METHODS LMLR was prepared and characterized by particle size, potential, and microscopy, and a rat model of acute blunt injury of gastrocnemius muscle was established. After treatments with resveratrol or LMLR, the therapeutic effects were evaluated by hematoxylin-eosin (HE) staining. The expression of MHCIIB and vimentin in mRNA level was measured by real-time PCR. The expression of desmin and collagen I protein was assessed by immunohistochemistry. RESULTS LMLR showed regular cycle shape in a size of ~1000 nm. LMLR was negatively charged (-30 mV). The in vitro release of LMLR was close to 80% at 10 h and 90% at 48 h. Acute gastrocnemius muscle injury was established in rats and tissue recovery was observed after LMLR treatment as evidenced by HE staining, decreased expression of MHCIIB, and increased expression of vimentin. Moreover, LMLR treatment obviously facilitated desmin expression and reduced collagen I expression. CONCLUSIONS LMLR is effective in treating acute blunt injury of gastrocnemius muscle in rats.

Spermidine promotes nucleus pulposus autophagy as a protective mechanism against apoptosis and ameliorates disc degeneration.

Spermidine has therapeutic effects in many diseases including as heart diastolic function, myopathic defects and neurodegenerative disorders via autophagy activation. Autophagy has been found to mitigate cell apoptosis in intervertebral disc degeneration (IDD). Accordingly, we theorize that spermidine may have beneficial effects on IDD via autophagy stimulation. In this study, spermidine's effect on IDD was evaluated in tert-butyl hydroperoxide (TBHP)-treated nucleus pulposus cells of SD rats in vitro as well as in a puncture-induced rat IDD model. We found that autophagy was actuated by spermidine in nucleus pulposus cells. In addition, spermidine treatment weakened the apoptotic effects of TBHP in nucleus pulposus cells. Spermidine increased the expression of anabolic proteins including Collagen-II and aggrecan and decreased the expression of catabolic proteins including MMP13 and Adamts-5. Additionally, autophagy blockade using 3-MA reversed the beneficial impact of spermidine against nucleus pulposus cell apoptosis. Autophagy was thus important for spermidine's therapeutic effect on IDD. Spermidine-treated rats had an accentuated T2-weighted signal and a diminished histological degenerative grade than vehicle-treated rats, showing that spermidine inhibited intervertebral disc degeneration in vivo. Thus, spermidine protects nucleus pulposus cells against apoptosis through autophagy activation and improves disc, which may be beneficial for the treatment of IDD.

FGF1 improves functional recovery through inducing PRDX1 to regulate autophagy and anti-ROS after spinal cord injury.

Fibroblast growth factor 1 (FGF1) is thought to exert protective and regenerative effects on neurons following spinal cord injury (SCI), although the mechanism of these effects is not well understood. The use of FGF1 as a therapeutic agent is limited by its lack of physicochemical stability and its limited capacity to cross the blood-spinal cord barrier. Here, we demonstrated that overexpression of FGF1 in spinal cord following SCI significantly reduced tissue loss, protected neurons in the ventricornu, ameliorated pathological morphology of the lesion, dramatically improved tissue recovery via neuroprotection, and promoted axonal regeneration and remyelination both in vivo and in vivo. In addition, the autophagy and the expression levels of PRDX1 (an antioxidant protein) were induced by AAV-FGF1 in PC12 cells after H2 O2 treatment. Furthermore, the autophagy levels were not changed in PRDX1-suppressing cells that were treated by AAV-FGF1. Taken together, these results suggest that FGF1 improves functional recovery mainly through inducing PRDX1 expression to increase autophagy and anti-ROS activity after SCI.

Lithium chloride contributes to blood-spinal cord barrier integrity and functional recovery from spinal cord injury by stimulating autophagic flux.

Blood-spinal cord barrier (BSCB) disruption following spinal cord injury (SCI) significantly compromises functional neuronal recovery. Autophagy is a potential therapeutic target when seeking to protect the BSCB. We explored the effects of lithium chloride (LiCl) on BSCB permeability and autophagy-induced SCI both in a rat model of SCI and in endothelial cells subjected to oxygen-glucose deprivation. We evaluated BSCB status using the Evans Blue dye extravasation test and measurement of tight junction (TJ) protein levels; we also assessed functional locomotor recovery. We detected autophagy-associated proteins in vivo and in vitro using both Western blotting and immunofluorescence staining. We found that, in a rat model of SCI, LiCl attenuated the elevation in BSCB permeability, improved locomotor recovery, and inhibited the degradation of TJ proteins including occludin and claudin-5. LiCl significantly induced the extent of autophagic flux after SCI by increasing LC3-II and ATG-5 levels, and abolishing p62 accumulation. In addition, a combination of LiCl and the autophagy inhibitor chloroquine not only partially eliminated the BSCB-protective effect of LiCl, but also exacerbated TJ protein degradation both in vivo and in vitro. Together, these findings suggest that LiCl treatment alleviates BSCB disruption and promotes locomotor recovery after SCI, partly by stimulating autophagic flux.

Dl-3-n-butylphthalide attenuates acute inflammatory activation in rats with spinal cord injury by inhibiting microglial TLR4/NF-κB signalling.

In this study, we examined the neuroprotective effects and anti-inflammatory properties of Dl-3-n-butylphthalide (NBP) in Sprague-Dawley (SD) rats following traumatic spinal cord injury (SCI) as well as microglia activation and inflammatory response both in vivo and in vitro. Our results showed that NBP improved the locomotor recovery of SD rats after SCI an significantly diminished the lesion cavity area of the spinal cord, apoptotic activity in neurons, and the number of TUNEL-positive cells at 7 days post-injury. NBP inhibited activation of microglia, diminished the release of inflammatory mediators, and reduced the upregulation of microglial TLR4/NF-κB expression at 1 day post-injury. In a co-culture system with BV-2 cells and PC12 cells, NBP significantly reduced the cytotoxicity of BV-2 cells following lipopolysaccharide (LPS) stimulation. In addition, NBP reduced the activation of BV-2 cells, diminished the release of inflammatory mediators, and inhibited microglial TLR4/NF-κB expression in BV-2 cells. Our findings demonstrate that NBP may have neuroprotective and anti-inflammatory properties in the treatment of SCI by inhibiting the activation of microglia via TLR4/NF-κB signalling.

Retinoic Acid Prevents Disruption of Blood-Spinal Cord Barrier by Inducing Autophagic Flux After Spinal Cord Injury.

Spinal cord injury (SCI) induces the disruption of the blood-spinal cord barrier (BSCB), which leads to infiltration of blood cells, inflammatory responses and neuronal cell death, with subsequent development of spinal cord secondary damage. Recent reports pointed to an important role of retinoic acid (RA), the active metabolite of the vitamin A, in the induction of the blood-brain barrier (BBB) during human and mouse development, however, it is unknown whether RA plays a role in maintaining BSCB integrity under the pathological conditions such as SCI. In this study, we investigated the BSCB protective role of RA both in vivo and in vitro and demonstrated that autophagy are involved in the BSCB protective effect of RA. Our data show that RA attenuated BSCB permeability and also attenuated the loss of tight junction molecules such as P120, ß-catenin, Occludin and Claudin5 after injury in vivo as well as in brain microvascular endothelial cells. In addition, RA administration improved functional recovery of the rat model of trauma. We also found that RA could significantly increase the expression of LC3-II and decrease the expression of p62 both in vivo and in vitro. Furthermore, combining RA with the autophagy inhibitor chloroquine (CQ) partially abolished its protective effect on the BSCB and exacerbated the loss of tight junctions. Together, our studies indicate that RA improved functional recovery in part by the prevention of BSCB disruption via the activation of autophagic flux after SCI.

Effects of gravel-sand and plastic film mulching on soil water and temperature retention in cold and arid regions without irrigation.

Gravel-sand mulch (GSM) and plastic film mulch (PFM) are important ways of farming in cold and arid regions without irrigation. Nevertheless, there has been a lack of studies of the system response to live weather conditions. To quantify the effects of GSM and PFM on soil moisture and temperature retention, in-situ monitoring experiments were carried out in the arid belt of central Ningxia, China, using continuous monitoring of the field soil water and meteorological conditions at a 30-mimute time-step under three treatments: a bare soil (CK), soil covered by a layer of GSM, and soil covered by GSM and a layer of plastic film (i.e., GSM + PFM). Results show that: (1) With a limited precipitation of 221 mm during the growing season, the average volumetric soil water content (SWC) in the top 30-cm soil layer was lowest for CK, medium high for GSM, and highest for GSM + PFM. Compared to CK, the soil water storage increased by 54 % under GSM and 75.2 % under GSM + PFM; (2) The most frequently occurring low-intensity rainfalls are more efficiently stored in soil under GSM + PFM; (3) Similarly, the soil temperature was significantly increased under GSM and GSM + PFM conditions. Compared to CK, the average soil temperature in the top 5-cm layer increased by 2.5 °C under GSM and 4.8 °C under GSM + PFM during the germination period, which had effectively extended the growing season for about 30 and 50 days, respectively; (4) Although dewfall is only 4 % of rainfall, the total number of dew day was more than twice that of rain day. Thus, dewfall is a more frequent and dependable source of water for native plants and animals. Our results demonstrate that the benefits of GSM and PFM can be applied globally where either insufficient rainfall or low temperatures are limiting factors.

Pulmonary Hypertension and the Risk of 30-Day Postoperative Pulmonary Complications after Gastrointestinal Surgical or Endoscopic Procedures: A Retrospective Propensity Score-Weighted Cohort Analysis.

Background: Pulmonary hypertension (PH) patients are at higher risk of postoperative complications. We analyzed the association of PH with 30-day postoperative pulmonary complications (PPCs). Methods: A single-center propensity score overlap weighting (OW) retrospective cohort study was conducted on 164 patients with a mean pulmonary artery pressure (mPAP) of >20 mmHg within 24 months of undergoing elective inpatient abdominal surgery or endoscopic procedures under general anesthesia and a control cohort (N = 1981). The primary outcome was PPCs, and the secondary outcomes were PPC sub-composites, namely respiratory failure (RF), pneumonia (PNA), aspiration pneumonia/pneumonitis (ASP), pulmonary embolism (PE), length of stay (LOS), and 30-day mortality. Results: PPCs were higher in the PH cohort (29.9% vs. 11.2%, p < 0.001). When sub-composites were analyzed, higher rates of RF (19.3% vs. 6.6%, p < 0.001) and PNA (11.2% vs. 5.7%, p = 0.01) were observed. After OW, PH was still associated with greater PPCs (RR 1.66, 95% CI (1.05-2.71), p = 0.036) and increased LOS (median 8.0 days vs. 4.9 days) but not 30-day mortality. Sub-cohort analysis showed no difference in PPCs between pre- and post-capillary PH patients. Conclusions: After covariate balancing, PH was associated with a higher risk for PPCs and prolonged LOS. This elevated PPC risk should be considered during preoperative risk assessment.

Incidence and risk factors of pain crisis after hematopoietic cell transplantation for sickle cell disease.

ABSTRACT: Vaso-occlusive episodes (VOC) or pain crises are the most common indications for hematopoietic cell transplantation (HCT) for sickle cell disease (SCD). Elimination of pain crisis after HCT is an important patient-centered outcome and may improve understanding of the natural history of pain syndromes in SCD. We examined deidentified records of 763 patients followed-up for a median of 36.7 months (range, 0.3-168.6 months), with 69.6% patient's age <18 years at HCT, 83.3% patient's Karnofsky-Lansky performance score (KPS) ≥90, overall survival 92.9%, event-free survival 72.4%, graft failure (GF) 22.4%, AGVHD 21.4%, CGVHD 27%, and pain crisis 8.65%. On unadjusted logistic regression, increased risk of pain crisis after HCT was observed in patient's aged >10 years at HCT (range, 11-17 years; OR, 9.43; 95% CI, 3.20-27.79; P < .0001), in age ≥18 years (OR, 16.62; 95% CI, 5.85-47.16; P < .0001), in those with history of pain crisis 2 years before HCT (OR, 13.16; 95% CI, 4.08-42.42; P < .0001), alternate donors (haploidentical [OR, 4.80; 95% CI, 2.48-9.31; P < .0001], unrelated matched [OR, 2.71; 95% CI, 1.23-5.97; P = .0132], and mismatched unrelated [OR, 3.19; 95% CI, 1.44-7.05; P = .0041], and those with GF (n = 41 [5.37%]; OR, 7.15; 95% CI, 4.20-12.18; P < .0001). Pain crisis was less frequent with KPS of ≥90 (OR, 0.31; 95% CI, 0.18-0.55; P < .0001). Multivariable logistic regression models confirmed age at HCT, KPS, graft type, donor type, history of VOC 2 years before HCT, and GF as independent predictors of pain crisis after HCT and generated predictive models and nomograms for pain crisis after HCT for SCD, which can support shared decision making.

Clinical and Functional Outcomes of Total Knee Arthroplasty in Sarcopenia: A Case-Control Retrospective Cohort Study.

BACKGROUND: The aim of the study was to investigate the relationship between sarcopenia and both clinical and functional outcome scores following total knee arthroplasty (TKA) performed for patients over 65 years of age. METHODS: We assessed patient demographics, preoperative health status, postoperative Knee Society Clinical (KSS-C) and Function (KSS-F) subscores, and perioperative complications for 180 patients with sarcopenia and 345 comparatively healthy patients at a mean of 12.0 months after surgery. Multivariate logistic regression analysis was performed to define whether sarcopenia was an independent risk factor for lower KSS-F and KSS-C subscores and peroperative complication rates. Patients with sarcopenia had lower mean body mass index, preoperative albumin, and preoperative hemoglobin levels (p < 0.01). RESULTS: Compared with the healthy control group, patients with sarcopenia had lower KSS-C (83.0 vs. 88.2, p < 0.01) and KSS-F (79.2 vs. 86.1, p < 0.01) subscores and increased postoperative complication rates (14.1% vs. 4.1%, p < 0.01). CONCLUSION: Patients with sarcopenia present with generally poorer preoperative health and this appears to be associated with lower patient-reported clinical and functional outcome scores. Complication rates were higher among patients with sarcopenia who were still determined to have adequate health status to support TKA. Most complications were limited and could be managed with supportive treatment.

TRPV4-mediated mitochondrial dysfunction induces pyroptosis and cartilage degradation in osteoarthritis via the Drp1-HK2 axis.

Osteoarthritis (OA) is an age-related chronic degenerative disease with complex pathophysiological mechanisms. Accumulating evidence indicates that nod-like receptor pyrin domain 3 (NLRP3) inflammasome-mediated pyroptosis of chondrocytes plays a crucial role in the OA progression. Transient Receptor Potential Vanilloid 4 (TRPV4), described as a calcium-permeable cation channel, isassociated with proinflammatory factors and pyroptosis. In this study, we studied the potential functions of TRPV4 in chondrocyte pyroptosis and cartilage degradation. We found that lipopolysaccharides(LPS)-induced mitochondrial reactive oxygen species (mtROS) accumulation aggravated chondrocyte pyroptosis and cartilage degeneration. TRPV4 induces dynamin-related protein 1 (Drp1) mitochondrial translocation through the Ca2+-calmodulin-dependent protein kinase II (CaMKII) signaling pathway, which subsequently caused the mitochondrial dysfunction (e.g., mPTP over opening; Δψm depolarization; ATP production decreased; mtROS accumulation), pyroptosis and extracellular matrix (ECM) degradation through hexokinase 2 (HK2) dissociation from mitochondrial membrane. Moreover, TRPV4 inhibition reversed Drp1-involved chondrocyte pyroptosis and cartilage degeneration in the anterior cruciate ligament transection (ACLT) mouse model. Our findings revealed the internal mechanisms underlying TRPV4 regulation in chondrocytes and its intrinsic therapeutic efficacy for OA.