The synergy of morphokinetic parameters and sHLA-G in cleavage embryo enhancing implantation rates.

Objective: This study aimed to assess the relationship between implantation and soluble HLA-G (sHLA-G) expression in cleavage embryo culture medium (ECM) in conjunction with early developmental kinetics determined by time-lapse imaging (TLI). Methods: A retrospective, single-center study was conducted involving 238 embryos from 165 patients who underwent Frozen-thawed embryo transfer (FET) using autologous oocytes, with either single or double embryo transfer. TLI morphokinetic parameters (t2, t3, t4, t5, t6, t7, t8, cc2, s2, cc3, s3) of embryos were analyzed, and sHLA-G levels in D3 ECM were measured using an enzyme-linked immunosorbent assay (ELISA). A hierarchical classification model was developed to categorize embryos into five groups (A, B, C, D, E). The correlation between sHLA-G levels, TLI classification of embryos, and embryo implantation was investigated to establish a non-invasive method for evaluating implantation potential. Multivariate logistic regression analysis was performed to identify potential influencing factors, and receiver operating characteristic (ROC) curves were used to evaluate the predictive value for implantation. Results: Multivariate unconditional logistic regression analysis indicated that TLI parameters t5 and s3 and sHLA-G level in ECM were independent risk factors affecting embryo implantation. The implantation rate decreased from TLI classification A to E. The proposed classification model effectively assessed the implantation potential of embryos. The implantation rate was higher in the sHLA-G positive group compared to the sHLA-G negative group (p < 0.001). The expression of sHLA-G in D3 ECM, combined with the TLI classification model, accurately evaluated the implantation potential of embryos with an AUC of 0.876. Conclusion: The integration of cleavage kinetics and embryonic sHLA-G expression could reliably identify embryos with a high likelihood of successful implantation.

Effectiveness and safety of interspinous spacer versus decompressive surgery for lumbar spinal stenosis: A meta-analysis of randomized controlled trials.

STUDY DESIGN: A meta-analysis of randomized controlled trials. OBJECTIVE: Our meta-analysis was conducted to investigate whether interspinous spacer (IS) results in better performance for patients with lumbar spinal stenosis (LSS) when compared with decompressive surgery (DS). BACKGROUND DATA: DS and IS are common surgeries for the treatment of LSS. However, controversy remains as to whether the IS is superior to DS. METHODS: We comprehensively searched PubMed, EMBASE, and Cochrane Central Register of Controlled Trials for prospective randomized controlled trials that compared IS versus DS for LSS. The retrieved results were last updated on July 30, 2023. RESULTS: Eight studies involving 852 individuals were included in the meta-analysis. The pooled data indicated that IS was superior to DS considering shorter operation time (P = .003), lower dural violation rate (P = .002), better Zurich Claudication Questionnaire Physical function score (P = .03), and smaller foraminal height decrease (P = .004), but inferior to DS considering the higher rate of reoperation (P < .0001). There was no significant difference between the 2 groups regarding hospital stay (P = .26), blood loss (P = .23), spinous process fracture (P = .09), disc height decrease (P = .87), VAS leg pain score (P = .43), VAS back pain score (P = .26), Oswestry Disability Index score (P = .08), and Zurich Claudication Questionnaire symptom severity (P = .50). CONCLUSIONS: In summary, we considered that IS had similar effects with DS in hospital stay, blood loss, spinous process fracture, disc height decrease, VAS score, Oswestry Disability Index score, and Zurich Claudication Questionnaire Symptom severity, and was better in some indices such as operation time, dural violation, Zurich Claudication Questionnaire Physical function, and foraminal height decrease than DS. However, due to the higher rate of reoperation in the IS group, we considered that both IS and DS were acceptable strategies for treating LSS. As a novel technique, further well-designed studies with longer-term follow-up are needed to evaluate the effectiveness and safety of IS.

Injectable hydrogel loaded with bilayer microspheres to inhibit angiogenesis and promote cartilage regeneration for repairing growth plate injury.

Introduction: The repair and regeneration of growth plate injuries using tissue engineering techniques remains a challenge due to large bone bridge formation and low chondrogenic efficiency. Methods: In this study, a bilayer drug-loaded microspheres was developed that contains the vascular endothelial growth factor (VEGF) inhibitor, Bevacizumab, on the outer layer and insulin-like growth factor-1 (IGF-1), a cartilage repair factor, on the inner layer. The microspheres were then combined with bone marrow mesenchymal stem cells (BMSCs) in the gelatin methacryloyl (GelMA) hydrogel to create a composite hydrogel with good injectability and biocompatibility. Results: The in vitro drug-release profile of bilayer microspheres showed a sequential release, with Bevacizumab released first followed by IGF-1. And this hydrogel simultaneously inhibited angiogenesis and promoted cartilage regeneration. Finally, in vivo studies indicated that the composite hydrogel reduced bone bridge formation and improved cartilage regeneration in the rabbit model of proximal tibial growth plate injury. Conclusion: This bilayer microsphere-based composite hydrogel with sequential controlled release of Bevacizumab and IGF-1 has promising potential for growth plate injury repair.

Lumbar endplate microfracture injury induces Modic-like changes, intervertebral disc degeneration and spinal cord sensitization - an in vivo rat model.

BACKGROUND CONTEXT: Endplate (EP) injury plays critical roles in painful IVD degeneration since Modic changes (MCs) are highly associated with pain. Models of EP microfracture that progress to painful conditions are needed to better understand pathophysiological mechanisms and screen therapeutics. PURPOSE: Establish in vivo rat lumbar EP microfracture model and assess crosstalk between IVD, vertebra and spinal cord. STUDY DESIGN/SETTING: In vivo rat EP microfracture injury model with characterization of IVD degeneration, vertebral remodeling, spinal cord substance P (SubP), and pain-related behaviors. METHODS: EP-injury was induced in 5 month-old male Sprague-Dawley rats L4-5 and L5-6 IVDs by puncturing through the cephalad vertebral body and EP into the NP of the IVDs followed by intradiscal injections of TNFα (n=7) or PBS (n=6), compared with Sham (surgery without EP-injury, n=6). The EP-injury model was assessed for IVD height, histological degeneration, pain-like behaviors (hindpaw von Frey and forepaw grip test), lumbar spine MRI and µCT, and spinal cord SubP. RESULTS: Surgically-induced EP microfracture with PBS and TNFα injection induced IVD degeneration with decreased IVD height and MRI T2 signal, vertebral remodeling, and secondary damage to cartilage EP adjacent to the injury. Both EP injury groups showed MC-like changes around defects with hypointensity on T1-weighted and hyperintensity on T2-weighted MRI, suggestive of MC type 1. EP injuries caused significantly decreased paw withdrawal threshold, reduced axial grip, and increased spinal cord SubP, suggesting axial spinal discomfort and mechanical hypersensitivity and with spinal cord sensitization. CONCLUSIONS: Surgically-induced EP microfracture can cause crosstalk between IVD, vertebra, and spinal cord with chronic pain-like conditions. CLINICAL SIGNIFICANCE: This rat EP microfracture model was validated to induce broad spinal degenerative changes that may be useful to improve understanding of MC-like changes and for therapeutic screening.

Lumbar endplate microfracture injury induces Modic-like changes, intervertebral disc degeneration and spinal cord sensitization - An In Vivo Rat Model.

BACKGROUND CONTEXT : Endplate (EP) injury plays critical roles in painful IVD degeneration since Modic changes (MCs) are highly associated with pain. Models of EP microfracture that progress to painful conditions are needed to better understand pathophysiological mechanisms and screen therapeutics. PURPOSE : Establish in vivo rat lumbar EP microfracture model with painful phenotype. STUDY DESIGN/SETTING : In vivo rat study to characterize EP-injury model with characterization of IVD degeneration, vertebral bone marrow remodeling, spinal cord sensitization, and pain-related behaviors. METHODS : EP-driven degeneration was induced in 5-month-old male Sprague-Dawley rats L4-5 and L5-6 IVDs through the proximal vertebral body injury with intradiscal injections of TNFα (n=7) or PBS (n=6), compared to Sham (surgery without EP-injury, n=6). The EP-driven model was assessed for IVD height, histological degeneration, pain-like behaviors (hindpaw von Frey and forepaw grip test), lumbar spine MRI and µCT analyses, and spinal cord substance P (SubP). RESULTS : EP injuries induced IVD degeneration with decreased IVD height and MRI T2 values. EP injury with PBS and TNFα both showed MC type1-like changes on T1 and T2-weighted MRI, trabecular bone remodeling on µCT, and damage in cartilage EP adjacent to the injury. EP injuries caused significantly decreased paw withdrawal threshold and reduced grip forces, suggesting increased pain sensitivity and axial spinal discomfort. Spinal cord dorsal horn SubP was significantly increased, indicating spinal cord sensitization. CONCLUSIONS : EP microfracture can induce crosstalk between vertebral bone marrow, IVD and spinal cord with chronic pain-like conditions. CLINICAL SIGNIFICANCE : This rat EP microfracture model of IVD degeneration was validated to induce MC-like changes and pain-like behaviors that we hope will be useful to screen therapies and improve treatment for EP-drive pain.

Spinal Cord Sensitization and Spinal Inflammation from an In Vivo Rat Endplate Injury Associated with Painful Intervertebral Disc Degeneration.

Intervertebral disc (IVD) degeneration with Modic-like changes is strongly associated with pain. Lack of effective disease-modifying treatments for IVDs with endplate (EP) defects means there is a need for an animal model to improve understanding of how EP-driven IVD degeneration can lead to spinal cord sensitization. This rat in vivo study determined whether EP injury results in spinal dorsal horn sensitization (substance P, SubP), microglia (Iba1) and astrocytes (GFAP), and evaluated their relationship with pain-related behaviors, IVD degeneration, and spinal macrophages (CD68). Fifteen male Sprague Dawley rats were assigned into sham or EP injury groups. At chronic time points, 8 weeks after injury, lumbar spines and spinal cords were isolated for immunohistochemical analyses of SubP, Iba1, GFAP, and CD68. EP injury most significantly increased SubP, demonstrating spinal cord sensitization. Spinal cord SubP-, Iba1- and GFAP-immunoreactivity were positively correlated with pain-related behaviors, indicating spinal cord sensitization and neuroinflammation play roles in pain responses. EP injury increased CD68 macrophages in the EP and vertebrae, and spinal cord SubP-, Iba1- and GFAP-ir were positively correlated with IVD degeneration and CD68-ir EP and vertebrae. We conclude that EP injuries result in broad spinal inflammation with crosstalk between spinal cord, vertebrae and IVD, suggesting that therapies must address neural pathologies, IVD degeneration, and chronic spinal inflammation.

Effects of high tibial osteotomy combined with arthroscopy on local inflammation degree and gait activity index in patients with medial knee osteoarthritis.

Objective: To analyze the effects of high tibial osteotomy (HTO) combined with arthroscopy on the degree of inflammation and the gait activity indexes of patients with medial knee osteoarthritis. Methods: We collected the records of 112 patients with medial knee osteoarthritis treated in the Department of Orthopedics of our hospital from June 2019 to June 2021. We divided the data into two groups: the control group included those of 54 patients who had received simple HTO and the observation group included those of 58 patients who had undergone HTO combined with arthroscopy. We assessed clinical efficacies, degrees of inflammation and gait activity indexes of the two groups to compare treatment outcomes. Results: The percentage of excellent and good outcomes in the observation group (89.66%) was significantly higher than that in the control group (66.67%; p < 0.05). One year after the operation, the serum and synovial fluid levels of interleukin-1ß (IL-1ß), interleukin-6 (IL-6) and interleukin-17 (IL-17) in the observation group were significantly lower than those in the control group (p < 0.05). Moreover, the double support phase was significantly lower in the observation group than in the control group, while the step length, speed and frequency were significantly higher in the observation group than in the control group (p < 0.05). Conclusions: HTO combined with Arthroscopy in patients with medial knee osteoarthritis improves the curative effect and the degree of inflammation, and it promotes the recovery of gait activity indices.

Risk, Prognostic Factors and Nomograms for Bone Metastasis in Young Females with Breast Cancer: A Large Cohort Retrospective Study.

Purpose: To determine the incidence of bone metastasis (BM) in young female patients with breast cancer (BC) and develop 2 robust nomograms for BM in young female patients with BC. Methods: We searched and downloaded the data from young (age ≤40 years) female patients with bone cancer from the Surveillance, Epidemiology, and End Results (SEER) database between 2010 and 2015. Univariate and multivariate analyses were performed to screen the potential diagnostic variables and prognostic factors for BM. The diagnostic and prognostic nomograms were generated and evaluated by the area under the receiver operating characteristic (ROC) curve (AUC), calibration curves, and decision curve analysis (DCA). Results: A total of 13 347 young female patients with BC were identified; of these, 462 were initially diagnosed as having BM. The independent risk factors for BM in young female patients with BC were tumor size, BC subtype, American Joint Committee on Cancer (AJCC) T stage, AJCC N stage, age and marital status. The independent prognostic factors in these patients were tumor size, subtype, surgery performed, lung metastasis, liver metastasis and brain metastasis. The AUC values of the diagnostic nomogram were 0.803 (95% CI; 0.795-0.811) and 0.813 (95% CI; 0.800-0.825) in the training and validation cohorts, respectively. The time-dependent AUC values of prognostic nomogram were 0.850, 0.853, and 0.824 at 2, 3 and 4 years in the training cohort, and also >0.700 in the validation cohort. For both nomograms, the discrimination was higher than all independent variables. Calibration curve and decision curve analysis (DCA) indicated that both nomograms had favorable calibration and clinical utilization. Finally, a risk stratification system was generated and the 3 risk subgroups showed significantly distinct prognoses. Conclusions: A total of 2 nomograms were developed to assess the risk for and in prognosis of young female patients with BC with BM (BCBM).

Discovery of Orally Bioavailable N-Benzylpiperidinol Derivatives as Potent and Selective USP7 Inhibitors with In Vivo Antitumor Immunity Activity against Colon Cancer.

USP7 emerges as a potential therapeutic target for cancers, as it plays an important role in the development of tumorigenesis by stabilizing multiple cancer-relevant proteins. Nevertheless, the discovery of drug-like USP7 inhibitors remains challenging. Herein, we report a series of N-benzylpiperidinol derivatives as potent and selective USP7 inhibitors (e.g., X20 and X26: IC50 = 7.6 and 8.2 nM), whose binding modes were revealed by crystallographic studies to be distinct from the known N-acylpiperidinol USP7 inhibitors. Among them, X36 with good oral PK profiles (rat: F = 40.8% and T1/2 = 3.5 h) exhibited significant antitumor efficacy in the MC38 colon cancer syngeneic mouse model, at least partly through upregulating the tumor infiltration of CD8+ T, NK, and NKT cells and downregulating that of Tregs and MDSCs. These findings may further pave the way for the development of USP7 inhibitors as novel cancer immunotherapy drugs.

Development and Validation of Nomograms to Assess Risk Factors and Overall Survival Prediction for Lung Metastasis in Young Patients with Osteosarcoma: A SEER-Based Study.

Background: To establish two nomograms to quantify the diagnostic factors of lung metastasis (LM) and their role in assessing prognosis in young patients with LM osteosarcoma. Methods: A total of 618 osteosarcoma young patients from 2010 to 2015 were included from the Surveillance, Epidemiology, and End Results (SEER) database. Another 131 patients with osteosarcoma from local hospitals were also collected as an external validation set. Patients were randomized into training sets (n = 434) and validation sets (n = 184) with a ratio of 7:3. Univariate and multivariate logistic regression analyses were used to identify the risk factor for LM and were used to construct the nomogram. Risk variables for the overall survival rate of patients with LM were evaluated by Cox regression. Another nomogram was also constructed to predict survival rates. The results were validated using bootstrap resampling and retrospective research on 131 osteosarcoma young patients from 2010 to 2019 at three local hospitals. Results: There were 114 (18.45%) patients diagnosed as LM at initial diagnosis. The multivariate logistic regression analysis suggested that T stage, N stage, and bone metastasis were independent risk factors for LM in newly diagnosed young osteosarcoma patients (P < 0.001). The ROC analysis revealed that area under the curve (AUC) values were 0.751, 0.821, and 0.735 in the training set, internal validation set, and external validation set, respectively, indicating good predictive discrimination. The multivariate Cox proportional hazard regression analysis suggested that age, surgery, chemotherapy, primary site, and bone metastasis were prognostic factors for young osteosarcoma patients with LM. The time-dependent ROC curves showed that the AUCs for predicting 1-year, 2-year, and 3-year survival rates were 0.817, 0.792, and 0.815 in the training set and 0.772, 0.807, and 0.804 in the internal validation set, respectively. As for the external validation set, the AUCs for predicting 1-year, 2-year, and 3-year survival rates were 0.787, 0.818, and 0.717. Conclusions: The nomograms can help clinicians strengthen their personal decision-making and can improve the prognosis of osteosarcoma patients.

Establishment of the diagnostic and prognostic nomograms for pancreatic cancer with bone metastasis.

Bone metastasis (BM) is rare in patients with pancreatic cancer (PC), but often neglected at the initial diagnosis and treatment. Bone metastasis is associated with a worse prognosis. This study was aimed to perform a large data analysis to determine the predictors and prognostic factors of BM in PC patients and to develop two nomograms to quantify the risks of BM and the prognosis of PC patients with BM. In the present study, we reviewed and collected the data of patients who were diagnosed as PC from 2010 to 2015 in the Surveillance, Epidemiology, and End Results (SEER) database. Univariate and multivariate logistic regression analyses were used together to screen and validate the risk factors for BM in PC patients. The independent prognostic factors for PC patients with BM were identified by Cox regression analysis. Finally, two nomograms were established via calibration curves, receiver operating characteristic (ROC) curve, and decision curve analysis (DCA). This study included 16,474 PC patients from the SEER database, and 226 of them were diagnosed with BM. The risk factors of BM for PC patients covered age, grade, T stage, N stage, tumor size, and primary site. The independent prognostic factors for PC patients with BM included age, race, grade, surgery, and lung metastasis. The AUC of the diagnostic nomogram was 0.728 in the training set and 0.690 in the testing set. In the prognostic nomogram, the AUC values of 6/12/18 month were 0.781/0.833/0.849 in the training set and 0.738/0.781/0.772 in the testing set. The calibration curve and DCA furtherly indicated the satisfactory clinical consistency of the nomograms. These nomograms could be accurate and personalized tools to predict the incidence of BM in PC patients and the prognosis of PC patients with BM. The nomograms can help clinicians make more personalized and effective treatment choices.

Facile Synthesis of Liquid Crystal Dimers Bridged with a Phosphonic Group.

Liquid crystal (LC) dimers with well-defined composition and structure arouse broad attentions for their exhibiting LC properties beyond conventional low molar mass mesogens and serving as fascinating model compounds for LC polymers. Here in this work, a series of LC dimers bridged with a phosphonic group have been synthesized through a facile free radical mediated addition reaction between hypophosphorous acid and vinyl terminated cyanobiphenyl mesogens with variant length alkyl spacers. In addition, two esterified derivatives and a group of mono-addition homologues with a terminal phosphonic acid group have also been prepared for comparison study. All the newly synthesized compounds exhibit monotropic nematic (N) phase with typical schlieren textures except for the LC dimer with the longest eleven-methylene spacer, which surprisingly shows twist-bend nematic (NTB ) phase directly from the isotropic state upon cooling. Moreover, the thermal transition properties such as the nematic-isotropic transition temperatures and associated entropy changes of the series LC dimers display a modest odd-even effect. Furthermore, both the LC dimers and the mono-addition homologues in N phase are quite easy to achieve homeotropic alignment upon annealing thanks to the supramolecular interactions between the introduced phosphonic acid group and the hydroxylated glass surface. This work thus provides a novel synthesis strategy for a class of LC materials bridged with a phosphonic acid group prone to further functionalization, which may serve as promising vertical alignment agents and pave the way for developing a new kind of functionalized LC materials of NTB phase.

Nonradical electron transfer-based peroxydisulfate activation by a Mn-Fe bimetallic oxide derived from spent alkaline battery for the oxidation of bisphenol A.

Mn-Fe bimetallic oxide has been employed as an outstanding peroxydisulfate (PDS) activator, but the underlying mechanism is still controversial. In this work, Mn0.27FeO4.55 (MFBO) was synthesized using the recovered waste alkaline battery and its catalytic activity and mechanism for PDS activation were explored in detail. Results show that MFBO exhibited a higher catalytic activity than the individual single metal oxides (FeOx and Mn2O3) due to the synergistic effect between Fe and Mn elements. The removal efficiency of bisphenol A (BPA) with an initial concentration of 10 mg/L reached 97.8% within 90 min in the presence of 0.5 g/L MFBO and 2.0 mM PDS. Moreover, the MFBO maintained high stability and reusability even after being recycled for five times. With the aid of a series of experiments and ex-situ/in-situ characterizations, a non-radical PDS activation mechanism was proposed, in which organic contaminants would be oxidized through a direct electron transfer pathway mediated by the metastable reactive complexes (MFBO-PDS*). Notably, the MFBO/PDS system revealed selective oxidation towards different organic pollutants and the reaction rates were closely related to their structures and properties. The research provided an effective alternation process for application of the waste battery, as well as developed a novel perspective for removal of recalcitrant aqueous contaminants through a nonradical mechanism.

TBC1D2 Promotes Ovarian Cancer Metastasis via Inducing E-Cadherin Degradation.

Background: Ovarian cancer (OC) is the most lethal gynecological malignancy worldwide. Increasing evidence indicates that TBC domain family is implicated in various cellular events contributing to initiation and development of different cancers, including OC. However, the role of TBC1D2, a crucial member of TBC domain family, remains unclear in OC. Methods: IHC and qRT-PCR were employed to determine TBC1D2 expression in OC tissues and cells. In vitro and in vivo assays involving proliferation, migration, invasion were performed to explore the role of TBC1D2 in OC development. The underlying mechanism by which TBC1D2 promotes OC metastasis were elucidated using bioinformatics analysis, western blotting and co-immunoprecipitation. Results: Upregulation of TBC1D2 was found in OC and was associated with a poor prognosis. Meanwhile, TBC1D2 promoted OC cell proliferation, migration, and invasion in vitro and facilitated tumor growth and metastasis in vivo. Moreover, TBC1D2 contributed to OC cell invasion by E-cadherin degradation via disassembling Rac1-IQGAP1 complex. In addition, miR-373-3p was screened out and identified to inhibit OVCAR3 invasion via negative regulation of TBC1D2. Conclusion: Our findings indicated that TBC1D2 is overexpressed in OC and contributes to tumor metastasis via E-cadherin degradation. This study suggests that TBC1D2 may be an underlying therapeutic target for OC.

Ex vivo biomechanical evaluation of Acute lumbar endplate injury and comparison to annulus fibrosus injury in a rat model.

Back pain is often associated with intervertebral disc (IVD) degeneration, and IVD degeneration phenotypes are commonly characterized by annulus fibrosus (AF)-driven and endplate (EP)-driven phenotypes. Few studies of EP injury exist in animal models, even though clinical studies show EP lesions are strongly associated with IVD pathology and pain. This project established an ex-vivo rat lumbar EP injury model and characterized effects of EP injury on motion segment biomechanical properties, as compared to AF injury, a common way of inducing IVD degeneration. Lumbar motion segments (39 total vertebra-IVD-vertebra sections) assigned to Intact (L1/L2), AF injury and EP injury (L3/L4 and L5/L6 randomly selected), and biomechanically tested in axial tension-compression, stress-relaxation and torsional testing in pre-injury and post-injury conditions using a repeated-measures design. EP injury involved superior vertebra endplate puncture transcorporeally and obliquely. AF injury involved mid-line punctures anterior and bilaterally. Axial ROM, tensile stiffness, hysteresis, and neutral zone stiffness were significantly affected by EP injury but not AF injury. Torque range, torsional stiffness and torsional neutral zone stiffness were significantly affected by AF injury but not EP injury. Stress-relaxation fast time constant was decreased for EP injury. EP and AF injuries induced distinct biomechanical changes in lumbar motion segments with EP injury having the largest impact on axial biomechanical properties and AF injury most prominently affecting torsional properties. This study deepens the understanding of biomechanical mechanism of EP-driven low back pain and provides methods and biomechanical characterization for future in vivo studies.

TFAM loss induces nuclear actin assembly upon mDia2 malonylation to promote liver cancer metastasis.

The mechanisms underlying cancer metastasis remain poorly understood. Here, we report that TFAM deficiency rapidly and stably induced spontaneous lung metastasis in mice with liver cancer. Interestingly, unexpected polymerization of nuclear actin was observed in TFAM-knockdown HCC cells when cytoskeleton was examined. Polymerization of nuclear actin is causally linked to the high-metastatic ability of HCC cells by modulating chromatin accessibility and coordinating the expression of genes associated with extracellular matrix remodeling, angiogenesis, and cell migration. Mechanistically, TFAM deficiency blocked the TCA cycle and increased the intracellular malonyl-CoA levels. Malonylation of mDia2, which drives actin assembly, promotes its nuclear translocation. Importantly, inhibition of malonyl-CoA production or nuclear actin polymerization significantly impeded the spread of HCC cells in mice. Moreover, TFAM was significantly downregulated in metastatic HCC tissues and was associated with overall survival and time to tumor recurrence of HCC patients. Taken together, our study connects mitochondria to the metastasis of human cancer via uncovered mitochondria-to-nucleus retrograde signaling, indicating that TFAM may serve as an effective target to block HCC metastasis.

Mitochondrial fragmentation is crucial for c-Myc-driven hepatoblastoma-like liver tumors.

Hepatoblastoma is the most common liver cancer in children, and the aggressive subtype often has a poor prognosis and lacks effective targeted therapy. Although aggressive hepatoblastoma (HB) is often accompanied by abnormally high expression of the transcription factor c-Myc, the underlying mechanism remains unclear. In this study, we found that mitochondrial fragmentation was enhanced by c-Myc overexpression in human aggressive HB tissues and was associated with poor prognosis. Then, a mouse model resembling human HB was established via hydrodynamic injection of c-Myc plasmids. We observed that liver-specific knockout of the mitochondrial fusion molecule MFN1 or overexpression of mitochondrial fission molecule DRP1 promoted the occurrence of c-Myc-driven liver cancer. In contrast, when MFN1 was overexpressed in the liver, tumor formation was delayed. In vitro experiments showed that c-Myc transcriptionally upregulated the expression of DRP1 and decreased MFN1 expression through upregulation of miR-373-3p. Moreover, enhanced mitochondrial fragmentation significantly promoted aerobic glycolysis and the proliferation of HB cells by significantly increasing reactive oxygen species (ROS) production and activating the RAC-alpha serine/threonine-protein kinase (AKT)/mammalian target of rapamycin (mTOR) and nuclear factor κB (NF-κB) pathways. Taken together, our results indicate that c-Myc-mediated mitochondrial fragmentation promotes the malignant transformation and progression of HB by activating ROS-mediated multi-oncogenic signaling.

Mesenchymal Stem Cell-Derived Extracellular Vesicles Inhibit Osteoporosis via MicroRNA-27a-Induced Inhibition of DKK2-Mediated Wnt/ß-Catenin Pathway.

Osteoporosis (OP) is a systemic skeletal disease that promotes bone fragility and the risk of fractures. Recent studies have shown the relevance of microRNAs (miRNAs) in the development of OP. This study aimed to evaluate the possible mechanisms of action underlying miR-27a loaded by mesenchymal stem cell (MSC)-derived extracellular vesicles (MSC-EVs) in OP. Serum samples from OP patients and normal controls were collected for miRNA microarray analysis. The expression of filtered miRNA was upregulated in osteoblasts (OB) and osteoclasts (OCs) for biological activity assessment. After developing OP mice using ovariectomy (OVX) and confirming OP, the miR-27a expression level was upregulated in mice by MSC-EV application. Dual-luciferase assays were conducted to validate the relationship between miR-27a and DKK2 expression. The poor expression of miR-27a was observed in patients with OP. miR-27a increased the expression of OB markers, the number of ALP-positive cells, and the number of calcium nodules in OCs. In OVX mice, miR-27a increased bone density, improved bone structure damage recovery, decreased the levels of bone resorption markers, and decreased OC number. miR-27a transmitted by MSC-EVs interacted with DKK2. MSC-EVs exerted the same protective effects as miR-27a on OP, whereas miR-27a inhibitor abolished the attenuating effects of MSC-EVs. In contrast, DKK2 depletion reversed the stimulatory effects of the miR-27a inhibitor on OP. The Wnt/ß-catenin pathway was activated upon MSC-EV application and DKK2 silencing and was impaired upon the downregulation of the expression of miR-27a. MSC-EVs are effective in preventing mouse OP. This mechanism is mediated by the miR-27a/DKK2/Wnt/ß-catenin signaling pathway.

NEP1-40-overexpressing Neural Stem Cells Enhance Axon Regeneration by Inhibiting Nogo-A/NgR1 Signaling Pathway.

BACKGROUND: Nogo-66 antagonistic peptide (NEP1-40) offers the potential to improve spinal cord injury (SCI). OBJECTIVE: To explore the effect of NEP1-40 overexpression on neural stem cells (NSCs) regulating the axon regeneration of injured neurons. METHODS: We isolated NSCs from brain tissues of pregnant rat fetuses and used Nestin immunofluorescence to identify them. The NEP1-40 overexpressing NSCs were constructed by transfection with the NEP1-40-overexpressing vector. The expression of NSCs differentiation associated markers, including Tuj-1, GFAP, Oligo2, and MBP, were detected by RT-PCR, western blotting, and immunofluorescence. NeuN immunofluorescence staining was used to measure the number of neurons. And western blotting was used to detect the phosphorylation levels of LIMK1/2, cofilin, and MLC-2 and the protein levels of GAP-43, MAP-2, and APP. RESULTS: The NEP1-40 overexpression promoted the expression level of Tuj-1, Oligo2, and MBP, and increased the number of Tuj-1, Oligo2, and MBP positive cells. NEP1-40-overexpressing NSCs (NEP-NSCs) improved NeuN positive cells of co-culture with injured neurons. And NEP-NSCs also increased the protein levels of axon regeneration indicators (GAP-43, MAP-2) and decreased APP protein level. In addition, the phosphorylation level of LIMK1/2, cofilin, and MLC-2 were markedly decreased in NEP-NSCs. CONCLUSION: NEP1-40 overexpression enhanced the ability of NSCs differentiation into neurons and promoted axon regeneration by inhibiting the Nogo-A/NgR1 signaling pathway. This study provides an alternative gene modified transplantation NSCs for the SCI treatment.

Mitochondrial transcription factor A plays opposite roles in the initiation and progression of colitis-associated cancer.

BACKGROUND: Mitochondria are key regulators in cell proliferation and apoptosis. Alterations in mitochondrial function are closely associated with inflammation and tumorigenesis. This study aimed to investigate whether mitochondrial transcription factor A (TFAM), a key regulator of mitochondrial DNA transcription and replication, is involved in the initiation and progression of colitis-associated cancer (CAC). METHODS: TFAM expression was examined in tissue samples of inflammatory bowel diseases (IBD) and CAC by immunohistochemistry. Intestinal epithelial cell (IEC)-specific TFAM-knockout mice (TFAM△IEC ) and colorectal cancer (CRC) cells with TFAM knockdown or overexpression were used to evaluate the role of TFAM in colitis and the initiation and progression of CAC. The underlying mechanisms of TFAM were also explored by analyzing mitochondrial respiration function and biogenesis. RESULTS: The expression of TFAM was downregulated in active IBD and negatively associated with the disease activity. The downregulation of TFAM in IECs was induced by interleukin-6 in a signal transducer and activator of transcription 3 (STAT3)/miR-23b-dependent manner. In addition, TFAM knockout impaired IEC turnover to promote dextran sulfate sodium (DSS)-induced colitis in mice. Of note, TFAM knockout increased the susceptibility of mice to azoxymethane/DSS-induced CAC and TFAM overexpression protected mice from intestinal inflammation and colitis-associated tumorigenesis. By contrast, TFAM expression was upregulated in CAC tissues and contributed to cell growth. Furthermore, it was demonstrated that ß-catenin induced the upregulation of TFAM through c-Myc in CRC cells. Mechanistically, TFAM promoted the proliferation of both IECs and CRC cells by increasing mitochondrial biogenesis and activity. CONCLUSIONS: TFAM plays a dual role in the initiation and progression of CAC, providing a novel understanding of CAC pathogenesis.