Astrocytic Epoxyeicosatrienoic Acid Signaling in the Medial Prefrontal Cortex Modulates Depressive-like Behaviors.

Major depressive disorder is the most common mental illness. Mounting evidence indicates that astrocytes play a crucial role in the pathophysiology of depression; however, the underlying molecular mechanisms remain elusive. Compared with other neuronal cell types, astrocytes are enriched for arachidonic acid metabolism. Herein, we observed brain-region-specific alterations of epoxyeicosatrienoic acid (EET) signaling, which is an arachidonic acid metabolic pathway, in both a mouse model of depression and postmortem samples from patients with depression. The enzymatic activity of soluble epoxide hydrolase (sEH), the key enzyme in EET signaling, was selectively increased in the mPFC of susceptible mice after chronic social defeated stress and was negatively correlated with the social interaction ratio, which is an indicator of depressive-like behavior. The specific deletion of Ephx2 (encode sEH) in adult astrocytes induced resilience to stress, whereas the impaired EET signaling in the mPFC evoked depressive-like behaviors in response to stress. sEH was mainly expressed on lysosomes of astrocytes. Using pharmacological and genetic approaches performed on C57BL/6J background adult male mice, we found that EET signaling modulated astrocytic ATP release in vitro and in vivo Moreover, astrocytic ATP release was required for the antidepressant-like effect of Ephx2 deletion in adult astrocytes. In addition, sEH inhibitors produced rapid antidepressant-like effects in multiple animal models of depression, including chronic social defeated stress and chronic mild stress. Together, our results highlight that EET signaling in astrocytes in the mPFC is essential for behavioral adaptation in response to psychiatric stress.SIGNIFICANCE STATEMENT Astrocytes, the most abundant glial cells of the brain, play a vital role in the pathophysiology of depression. Astrocytes secrete adenosine ATP, which modulates depressive-like behaviors. Notably, astrocytes are enriched for arachidonic acid metabolism. In the present study, we explored the hypothesis that epoxyeicosatrienoic acid signaling, an arachidonic acid metabolic pathway, modulates astrocytic ATP release and the expression of depressive-like behaviors. Our work demonstrated that epoxyeicosatrienoic acid signaling in astrocytes in the mPFC is essential for behavioral homeostatic adaptation in response to stress, and the extent of astrocyte functioning is greater than expected based on earlier reports.

Atractylenolide-III attenuates osteoarthritis by repolarizing macrophages through inactivating TLR4/NF-κB signaling.

BACKGROUND: As a common chronic musculoskeletal condition, osteoarthritis (OA) presently lacks particular treatment strategies. The aim of this study was to examine how AT-III therapies affected macrophage repolarity in order to slow down the advancement of OA. METHODS: RAW264.7 macrophages were polarized to M1 subtypes then administered with different concentrations of AT-III. Immunofluorescence, qRT-PCR and flow cytometry were used to assess the polarization of the macrophages. The mechanism of AT-III repolarize macrophages was evaluated by western blot. Furthermore, the effects of macrophage conditioned media (CM) on the migration, proliferation, and chondrogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) were investigated using CCK-8 assays, the scratch test, and alcian blue staining. The effects of macrophage CM on chondrocyte proliferation and degeneration were investigated using CCK-8 and qRT-PCR. In vivo micro-CT and histological observations were performed on rats with anterior cruciate ligament transection and partial medial meniscectomy, either with or without AT-III treatment. RESULTS: AT-III repolarized M1 macrophages to M2 phenotype. Mechanistically, AT-III reduced the expression of Toll-like receptor(TLR) 4 induced by lipopolysaccharide in RAW264.7 and lowered nuclear factor-κB (NF-κB) signaling molecules p-p65 and p-IκBα. The TLR4 agonist RS09 reversed the effects of AT-III on macrophage repolarization. AT-III-induced macrophages CM stimulated BMSCs migration, proliferation and chondrogenic differentiation. AT-III-treated macrophage CM promoted chondrocyte proliferation while inhibiting chondrocyte degeneration. In vivo, AT-III treatment alleviated the degree of synovitis, inhibited subchondral bone remodeling and reduced cartilage destruction in the rat OA model. CONCLUSIONS: AT-III attenuates OA by repolarizing macrophages through inactivating TLR4/NF-κB signaling. These data suggest that AT-III may be an effective therapeutic candidate for OA treatment.

STS loaded PCL-MECM based hydrogel hybrid scaffolds promote meniscal regeneration via modulating macrophage phenotype polarization.

Meniscus injury has a limited ability to heal itself and often results in the progression to osteoarthritis. After a meniscus injury, there is an obvious acute or chronic inflammatory response in the articular cavity, which is not conducive to tissue regeneration. M2 macrophages are involved in tissue repair and remodeling. Regenerative medicine strategies for tissue regeneration by enhancing the phenotypic ratio of M2 : M1 macrophages have been demonstrated in a variety of tissues. However, there are no relevant reports in the field of meniscus tissue regeneration. In this study, we confirmed that sodium tanshinone IIA sulfonate (STS) could transform macrophages from M1 to M2 polarization. STS protects meniscal fibrochondrocytes (MFCs) against the effects of macrophage conditioned medium (CM). Moreover, STS attenuates interleukin (IL)-1ß-induced inflammation, oxidative stress, apoptosis, and extracellular matrix (ECM) degradation in MFCs, possibly by inhibiting the interleukin-1 receptor-associated kinase 4 (IRAK4)/TNFR-associated factor 6 (TRAF6)/nuclear factor-kappaB (NF-κB) signaling pathway. An STS loaded polycaprolactone (PCL)-meniscus extracellular matrix (MECM) based hydrogel hybrid scaffold was fabricated. PCL provides mechanical support, the MECM based hydrogel provides a microenvironment conducive to cell proliferation and differentiation, and STS is used to drive M2 polarization and protect MFCs against the effects of inflammatory stimuli, thus providing an immune microenvironment conducive to regeneration. The results of subcutaneous implantation in vivo showed that hybrid scaffolds could induce M2 polarization in the early stage. In addition, the hybrid scaffolds seeded with MFCs could achieve good meniscus regeneration and chondroprotective effects in rabbits.

Expression and Clinical Significance of High-Mobility Group AT-hook 2 (HMGA2) in Osteosarcoma.

OBJECTIVE: Although high-mobility group AT-hook 2 (HMGA2) has been shown to have crucial roles in the pathogenesis and metastasis of various malignancies, its expression and significance in osteosarcoma remain unknown. Here we evaluate the expression, clinical prognostic value, and overall function of HMGA2 in osteosarcoma. METHODS: Sixty-nine osteosarcoma patient specimens within a tissue microarray (TMA) were analyzed by immunohistochemistry for HMGA2 expression. Demographics and clinicopathological information including age, gender, tumor location, metastasis, recurrence, chemotherapy response, follow-up time, and disease status were also collected. After validation of expression, we determined whether there was a correlation between HMGA2 expression and patient clinicopathology. HMGA2 expression was also evaluated in osteosarcoma cell lines and patient tissues by Western blot, we analyzed the expression of HMGA2 in the human osteosarcoma cell lines MG63, 143B, U2OS, Saos-2, MNNG/HOS, and KHOS. HMGA2-specific siRNA and clonogenic assays were then used to determine the effect of HMGA2 inhibition on osteosarcoma cell proliferation, growth, and chemosensitivity. RESULTS: HMGA2 expression was elevated in the osteosarcoma patient specimens and human osteosarcoma cell lines. HMGA2 was differentially expressed in human osteosarcoma cell lines. Specifically, a relatively high expression of HMGA2 was present in KHOS, MNNG/HOS, 143B and a relatively low expression was in MG63, U2OS as well as Saos-2. HMGA2 expression is correlated with metastasis and shorter overall survival. High HMGA2 expression is an independent predictor of poor osteosarcoma prognosis. There was no significant correlation between HMGA2 expression and the age, gender, or tumor site of the patient. HMGA2 expression is predominantly within the nucleus. The expression of HMGA2 also directly correlated to neoadjuvant chemoresistance. There was a significant reduction of HMGA2 expression in the siRNA transfection group. After the use of siRNA, the proliferation of osteosarcoma cells is decreased and the chemosensitivity of osteosarcoma cells is significantly increased. CONCLUSION: Our study supports HMGA2 as a potential prognostic biomarker and therapeutic target in osteosarcoma.

Assessing the Efficacy of the Early Rehabilitation Pathway in Combination with Morita Therapy after Hip and Knee Arthroplasty.

Total hip arthroplasty (THA) and total knee arthroplasty (TKA) are effective methods for the treatment of end-stage osteoarthritis. Furthermore, rehabilitation training and psychological interventions play significant roles in the recovery of hip and knee joint function after THA and TKA. A total of 46 patients who received hip replacement and knee replacement are equally divided into two groups, with the control group being prescribed routine rehabilitation intervention and the observation group prescribed an early rehabilitation pathway with Morita therapy intervention. According to the results, the observation group displayed a significantly decreased incidence of deep venous thrombosis, while simultaneously reducing the recovery time of lower limb function (P < 0.05), including straight leg raising time, walking time, and vertical knee flexion time. In addition, the treatment program demonstrates a significant ability to improve the joint function score, pain score, quality of life score, and range of motion score (P < 0.05). Moreover, serum D-dimer, fibrin degradation products (FDP), and femoral vein blood flow peak also are significantly reduced (P < 0.05). Therefore, we have determined that an early rehabilitation pathway combined with Morita therapy can effectively reduce stress pain, improve the recovery process of joint motor function, and reduce the incidence of thrombosis. However, an increased sample size would facilitate the confirmation of the safety and efficacy of the program. In addition, the overall financial expenditure and feasibility of the treatment need to be considered.

The Role of Zinc in Bone Mesenchymal Stem Cell Differentiation.

Zinc is an essential trace element for bone growth and bone homeostasis in the human body. Bone mesenchymal stem cells (BMSCs) are multipotent progenitors existing in the bone marrow stroma with the capability of differentiating along multiple lineage pathways. Zinc plays a paramount role in BMSCs, which can be spurred differentiating into osteoblasts, chondrocytes, or adipocytes, and modulates the formation and activity of osteoclasts. The expression of related genes also changed during the differentiation of various cell phenotypes. Based on the important role of zinc in BMSC differentiation, using zinc as a therapeutic approach for bone remodeling will be a promising method. This review explores the role of zinc ion in the differentiation of BMSCs into various cell phenotypes and outlines the existing research on their molecular mechanism.

A hippocampus dependent neural circuit loop underlying the generation of auditory mismatch negativity.

Extracting relevant information and transforming it into appropriate behavior, is a fundamental brain function, and requires the coordination between the sensory and cognitive systems, however, the underlying mechanisms of interplay between sensory and cognition systems remain largely unknown. Here, we developed a mouse model for mimicking human auditory mismatch negativity (MMN), a well-characterized translational biomarker for schizophrenia, and an index of early auditory information processing. We found that a subanesthetic dose of ketamine decreased the amplitude of MMN in adult mice. Using pharmacological and chemogenetic approaches, we identified an auditory cortex-entorhinal cortex-hippocampus neural circuit loop that is required for the generation of MMN. In addition, we found that inhibition of dCA1→MEC circuit impaired the auditory related fear discrimination. Moreover, we found that ketamine induced MMN deficiency by inhibition of long-range GABAergic projection from the CA1 region of the dorsal hippocampus to the medial entorhinal cortex. These results provided circuit insights for ketamine effects and early auditory information processing. As the entorhinal cortex is the interface between the neocortex and hippocampus, and the hippocampus is critical for the formation, consolidation, and retrieval of episodic memories and other cognition, our results provide a neural mechanism for the interplay between the sensory and cognition systems.

Increased EHHADH Expression Predicting Poor Survival of Osteosarcoma by Integrating Weighted Gene Coexpression Network Analysis and Experimental Validation.

Enoyl-CoA hydratase and 3-hydroxyacyl CoA dehydrogenase (EHHADH), a member of the 3-hydroxyacyl-CoA dehydrogenase family, were previously demonstrated to be involved in the tumorigenesis of various cancer types. This study is aimed at determining of the diagnostic and prognostic value of EHHADH in osteosarcoma (OS). The overexpression of EHHADH was found both in OS and also other sarcoma types, and according to the retrospective cohort study, the EHHADH level was related to the overall survival and disease-free survival of the OS patients. Furthermore, knockdown of EHHADH under the influence of EHHADH small interfering RNA significantly suppressed the proliferation ability of the tumor cells. Moreover, EHHADH overexpressed was found in human OS tissues. In summary, the progression of OS could be enhanced by EHHADH, which may be a potential diagnostic and prognostic biomarker for OS patients.

IRAK-4 in macrophages contributes to inflammatory osteolysis of wear particles around loosened hip implants.

TLRs recognizing PAMPS play a role in local immunity and participate in implant-associated loosening. TLR-mediated signaling is primarily regulated by IL-1 receptor associated kinase-M (IRAK-M) negatively and IRAK-4 positively. Our previous studies have proved that wear particles promote endotoxin tolerance in macrophages by inducing IRAK-M. However, whether IRAK-4 is involved in inflammatory osteolysis of wear particles basically, and the specific mechanism of IRAK-4 around loosened hip implants, is still unclear. IRAK-4 was studied in the interface membranes from patients in vivo and in particle-stimulated macrophages to clarify its role. Also, IL-1ß and TNF-α levels were measured after particle and LPS stimulation in macrophages with or without IRAK-4 silenced by siRNA. Our results showed that the interface membranes around aseptic and septic loosened prosthesis expressed more IRAK-4 compared with membranes from osteoarthritic patients. IRAK-4 in macrophages increased upon particle and LPS stimulation. In the former, IL-1ß and TNF-α levels were lower compared with those of LPS stimulation, and IRAK-4 siRNA could suppress production of pro-inflammatory cytokines. These findings suggest that besides IRAK-M, IRAK-4 also plays an important role in the local inflammatory reaction and contributes to prosthesis loosening.

Biomechanical study of medial meniscus after posterior horn injury: a finite element analysis.

We established an effective finite element model of knee joint for observation of stress and displacement of meniscus related changes after medial meniscus injury. Different types of medial meniscus injury can lead to varied meniscus stress and displacement changes. Stress and displacement concentration were found in fissure tip of meniscus tear compared to normal meniscus. The posterior horn injury of medial meniscus may initiate combined injury of medial meniscus posterior horn (MMPH) and that of medial meniscus body, and combined injury of MMPH and that of lateral meniscus anterior horn; fissure expansions regarding horizontal fissure, longitudinal fissure and grip-shaped fissure of MMPH were spotted.

Self-Inclusion Complexes Derived from Cyclodextrins: Synthesis and Characterization of 6(A),6(B)-Bis-O-[p-(allyloxy)phenyl]-Substituted beta-Cyclodextrins.

The syntheses, structures, and spectroscopic properties of 6(A),6(B)-bis-O-[p-(allyloxy)phenyl]-substituted beta-cyclodextrins have been investigated. Selective activation of the 6(A),6(B)-hydroxy groups was carried out by treating heptakis(2,3-di-O-methyl)-beta-cyclodextrin (1) with 2,4-dimethoxybenzene-1,5-disulfonyl chloride to give 6(A),6(B)-bissulfonate ester 2 in a yield of only 3%. This material was treated with sodium p-(allyloxy)phenoxide in DMF to form 6(A),6(B)-bis-O-[p-(allyloxy)phenyl]-heptakis(2,3-di-O-methyl)-beta-cyclodextrin (3), which had two isomers. One (3A) has the two p-(allyloxy)phenyl arms directed away from the cyclodextrin cavity, and the other (3B) has one of the p-(allyloxy)phenyl groups through the cavity to form a self-inclusion complex. When either 3A or 3B was treated with methyl iodide and sodium hydride, the resulting permethylated 6(A),6(B)-bis-O-[p-(allyloxy)phenyl]heptakis(2,3-di-O-methyl)-6(C),6(D),6(E),6(F),6(G)-penta-O-methyl-beta-cyclodextrin (4) was composed of two isomers, in which 4B is a self-inclusion complex. 3A and 3B also can be converted into a mixture of 3A and 3B in strong base but not when melted in the absence of base. 4A and 4B do not isomerize. Detailed 1D and 2D NMR spectroscopic studies were carried out to characterize the structures of these new compounds, and molecular mechanics techniques were used to explain the experimental facts.

[Analysis of the clinical effects of the modified halo pelvic frame for the treatment of severe scoliosis with rigidity].

OBJECTIVE: To evaluate the clinical curative effect of the modified Halo pelvic frame and surgery for the treatment of severe scoliosis with rigidity. METHODS: From January 2004 to May 2010,50 patients with severe scoliosis patients with rigidity were treated in our hospital. Twenty-three patients were male and 27 patients were female, with a mean age of 10.8 years old, ranging from 4 to 16 years. Twenty-four patients were congenital scoliosis and 26 patiens were idiopathic scoliosis. The mean body height were (152.1 +/- 11.1) cm and the average Cobb angle of scoliosis and kyphosis were (91.8 +/- 14.5) degrees and (69.5 +/- 14.0) degrees respectively. All the patients were treated with three-stages modified Halo pelvic traction, the second stage anterior release and the third stage posterior correction. The amount of correction was determined by measuring the change of body height, the Cobb angles and correction rate of scoliosis as well as kyphosis before and after the operation. RESULTS: The mean body height were correct to (158.5 +/- 10.5) cm. The average Cobb angle of scoliosis were correct to (30.8 +/- 7.9) degrees. The average Cobb angle of kyphosis were correct to (31.6 +/- 10.1) degrees. After the first stage, the average Cobb angle of scoliosis and kyphosis were changed with the mean of (30.4 +/- 6.6)% correction and (22.3 +/- 5.2)% respectively; after the second stage, the average Cobb angle of scoliosis and kyphosis were changed with the mean (26.7 -/+ 5.1)% correction and (21.2 -/+ 6.0)% respectively; the third stage, above data were (33.7 -/+ 7.2)% and (27.1 +/- 5.3)%. Correction rate of scoliosis and kyphosis were (66.5 +/- 7.2)% and (55.1 +/- 6.4)% respectively by the modified Halo pelvic frame traction and surgery. Body height, the Cobb angles and correction rate of scoliosis and kyphosis on radiographs were different in all stages (P<0.05). CONCLUSION: Operative complications of severe scoliosis with rigidity can be reduced and better deformity correction and trunk balance achieved by the modified Halo pelvic frame traction and surgery.