Luteolin regulating synthesis and catabolism of osteoarthritis chondrocytes via activating autophagy.

Osteoarthritis (OA) is a prevalent bone and joint disease characterized by degeneration. The dysregulation between chondrocyte synthesis and breakdown is a key factor in OA development. Targeting the degenerative changes in cartilage tissue degradation could be a potential treatment approach for OA. Previous research has established a strong link between autophagy and the regulation of chondrocyte functions. Activating autophagy has shown promise in mitigating cartilage tissue degeneration. Currently, osteoarthritis treatment primarily focuses on symptom management, as there is no definitive medication to stop disease progression. Previous studies have demonstrated that luteolin, a flavonoid present in Chinese herbal medicine, can activate autophagy and reduce the expression of MMP1 and ADAMTS-5. This study utilized an in vitro osteoarthritis model with chondrocytes stimulated by IL-1ß, treated with varying concentrations of luteolin. Treatment with luteolin notably increased the levels of synthesis factors Aggrecan and Collagen II, while decreasing the levels of decomposition factors MMP-1 and ADAMTS-5. Moreover, inhibition of autophagy by Chloroquine reversed the imbalances in chondrocyte activities induced by IL-1ß. In an in vivo model of knee osteoarthritis induced by medial meniscal instability (DMM), luteolin was administered as a therapeutic regimen. After 12 weeks, knee cartilage tissues from mice were analyzed. Immunofluorescence and immunohistochemical staining revealed a decrease in P62 expression and an increase in Beclin-1 in the cartilage tissues. Additionally, cartilage wear in the knee joints of mice was alleviated by safranin O and fast green staining. Our study findings underscore the significant role of luteolin in effectively rebalancing chondrocyte activities disrupted by IL-1ß. Our results strongly indicate that luteolin has the potential to be developed as a novel therapeutic agent for the treatment of osteoarthritis, offering promising prospects for future drug development.

Lipid metabolism: the potential targets for toxoplasmosis treatment.

Toxoplasmosis is a zoonosis caused by Toxoplasma gondii (T. gondii). The current treatment for toxoplasmosis remains constrained due to the absence of pharmaceutical interventions. Thus, the pursuit of more efficient targets is of great importance. Lipid metabolism in T. gondii, including fatty acid metabolism, phospholipid metabolism, and neutral lipid metabolism, assumes a crucial function in T. gondii because those pathways are largely involved in the formation of the membranous structure and cellular processes such as division, invasion, egress, replication, and apoptosis. The inhibitors of T. gondii's lipid metabolism can directly lead to the disturbance of various lipid component levels and serious destruction of membrane structure, ultimately leading to the death of the parasites. In this review, the specific lipid metabolism pathways, correlative enzymes, and inhibitors of lipid metabolism of T. gondii are elaborated in detail to generate novel ideas for the development of anti-T. gondii drugs that target the parasites' lipid metabolism.

Effect of the mitochondrial uncoupling agent BAM15 against the Toxoplasma gondii RH strain and Prugniaud strain.

BACKGROUND: Toxoplasmosis is a zoonotic disease caused by the infection of the protozoa Toxoplasma gondii (T. gondii), and safe and effective therapeutic drugs are lacking. Mitochondria, is an important organelle that maintains T. gondii survival, however, drugs targeting mitochondria are lacking. METHODS: The cytotoxicity of BAM15 was detected by CCK-8 and the in vitro effects of BAM15 was detected by qPCR, plaque assay and flow cytometry. Furthermore, the ultrastructural changes of T. gondii after BAM15 treatment were observed by transmission electron microscopy, and further the mitochondrial membrane potential (ΔΨm), ATP level and reactive oxygen species (ROS) of T. gondii after BAM15 treatment were detected. The pharmacokinetic experiments and in vivo infection assays were performed in mice to determine the in vivo effect of BAM15. RESULTS: BAM15 had excellent anti-T. gondii activity in vitro and in vivo with an EC50 value of 1.25 µM, while the IC50 of BAM15 in Vero cells was 27.07 µM. Notably, BAM15 significantly inhibited proliferation activity of T. gondii RH strain and Prugniaud strain (PRU), caused T. gondii death. Furthermore, BAM15 treatment induced T. gondii mitochondrial vacuolation and autolysis by TEM. Moreover, the decrease in ΔΨm and ATP level, as well as the increase in ROS production further confirmed the changes CONCLUSIONS: Our study identifies a useful T. gondii mitochondrial inhibitor, which may also serve as a leading molecule to develop therapeutic mitochondrial inhibitors in toxoplasmosis.'

RUNX2 prompts triple negative breast cancer drug resistance through TGF-ß pathway regulating breast cancer stem cells.

Triple-negative breast cancer (TNBC) stands out as the most aggressive subtype within the spectrum of breast cancer. The current clinical guidelines propose treatment strategies involving cytotoxic agents like epirubicin or paclitaxel. However, the emergence of acquired resistance frequently precipitates secondary tumor recurrence or the spread of metastasis. In recent times, significant attention has been directed toward the transcription factor RUNX2, due to its pivotal role in both tumorigenesis and the progression of cancer. Previous researches suggest that RUNX2 might be intricately linked to the development of resistance against chemotherapy, with its mechanism of action possibly intertwined with the signaling of TGF-ß. Nevertheless, the precise interplay between their effects and the exact molecular mechanisms underpinning chemoresistance in TNBC remain elusive. Therefore, we have taken a multifaceted approach from in vitro and in vivo experiments to validate the relationship between RUNX2 and TGF-ß and to search for their pathogenic mechanisms in chemoresistance. In conclusion, we found that RUNX2 affects chemoresistance by regulating cancer cell stemness through direct binding to TGF-ß, and that TGF-ß dually regulates RUNX2 expression. The important finding will provide a new reference for clinical reversal of the development of chemoresistance in breast cancer.

Identification of sulfur metabolism-related gene signature in osteoarthritis and TM9SF2's sustenance effect on M2 macrophages' phagocytic activity.

BACKGROUND: Osteoarthritis (OA) is a chronic and low-grade inflammatory disease associated with metabolism disorder and multiple cell death types in the synovial tissues. Sulfur metabolism has not been studied in OA. METHODS: First, we calculated the single sample gene set enrichment analysis score of sulfur metabolism-associated annotations (i.e., cysteine metabolism process, regulation of sulfur metabolism process, and disulfidptosis) between healthy and synovial samples from patients with OA. Sulfur metabolism-related differentially expressed genes (DEGs) were analyzed in OA. Least absolute shrinkage and selection operator COX regression were used to identify the sulfur metabolism-associated gene signature for diagnosing OA. Correlation and immune cell deconvolution analyses were used to explore the correlated functions and cell specificity of the signature gene, TM9SF2. TM9SF2's effect on the phagocytosis of macrophages M2 was analyzed by coculturing macrophages with IgG-coated beads or apoptotic Jurkat cells. RESULTS: A diagnostic six gene signature (i.e., MTHFD1, PDK4, TM9SF2, POU4F1, HOXA2, NCKAP1) was identified based on the ten DEGs, validated using GSE12021 and GSE1919 datasets. TM9SF2 was upregulated in the synovial tissues of OA at both mRNA and protein levels. The relationship between TM9SF2 and several functional annotations, such as antigen processing and presentation, lysosome, phagosome, Fcγ-mediated phagocytosis, and tyrosine metabolism, was identified. TM9SF2 and macrophages M2 were significantly correlated. After silencing TM9SF2 in THP-1-derived macrophages M2, a significantly reduced phagocytosis and attenuated activation of PLC-γ1 were observed. CONCLUSION: A sulfur metabolism-associated six-gene signature for OA diagnosis was constructed and upregulation of the phagocytosis-associated gene, TM9SF2, was identified. The findings are expected to deepen our understanding of the molecular mechanism underlying OA development and be used as potential therapeutic targets.

Genetic variant in SPAG16 is associated with the susceptibility of ACPA-positive rheumatoid arthritis possibly via regulation of MMP-3.

OBJECTIVES: In two previously published genome-wide association studies, a cluster of variants of sperm-associated antigen16 (SPAG16) were reported to be associated with the radiological progression rate of ACPA-positive rheumatoid arthritis (RA) patients from North American and Southern European ancestry. In this study, we aimed to investigate whether the reported RA-risk loci in SPAG16 are associated with the disease in the Chinese population and to further validate the functional role of the susceptible locus in RA tissues. METHODS: A total of 500 ACPA-positive RA patients and 1000 age-matched healthy subjects were recruited. Two SNPs of SPAG16, including rs7607479 (C/T) and rs6435818 (A/C), were genotyped, and the genotyping data were compared with chi-square test. Gene expression analysis was performed in synovial tissues obtained from 40 RA patients and 30 non-RA controls surgically treated for bone fracture. The tissue expression of SPAG16 and matrix metalloproteinase 3 (MMP-3) was compared between the two groups by the Student's t test. The relationship between serum indexes and mRNA expression of SPAG16 and MMP-3 were evaluated by Spearman's correlation analysis. RESULT: For rs7607479, the frequency of genotype TT was significantly higher in RA patients than in the controls (49.0% vs. 40.4%, p = 0.002). The RA patients were found to have significantly lower frequency of allele C than the controls (30.9% vs. 36.8%, p = 0.001). As for rs6435818, there was no significant difference of genotype or allele frequency between the two groups. The mRNA expression of MMP-3 was 1.63-fold higher in the RA patients than in the controls (p < 0.001). The expression of SPAG16 was comparable between the two groups (p = 0.43). The mRNA expression of MMP-3 was 1.39-fold higher in patients with genotype TT than in the patients with genotype CC (p = 0.006). The mRNA expression level of MMP-3 was significantly correlated with serum rheumatoid factor (r = 0.498, p < 0.001) and C-reactive protein (r = 0.272, p = 0.01), weakly correlated with erythrocyte sedimentation rate (r = 0.236, p = 0.09). CONCLUSIONS: We validated a common genetic risk factor in ACPA-positive patients with RA, which is associated with the tissue production of MMP-3 and disease progression. Further functional analysis into the role of rs7607479 in MMP-3 expression can shed new light on the genetic architecture of ACPA-positive RA.

Dehydrozingerone Alleviates Hyperalgesia, Oxidative Stress and Inflammatory Factors in Complete Freund's Adjuvant-Induced Arthritic Rats.

Purpose: Rheumatoid arthritis (RA) is a chronic autoimmune disease with severe inflammatory responses. Dehydrozingerone (DHZ) is a potent bioactive compound found in the rhizomes of Zingiber officinale, and it has been reported as an excellent anti-inflammatory and antioxidant agent. This study evaluated the anti-arthritic effects of DHZ in complete Freund's adjuvant (CFA)-induced arthritis. Methods: CFA administered rats were intragastrically treated with DHZ (100 mg/kg) for 28 days, and arthritis severity was assessed via body weight, arthritic score, paw edema and hyperalgesia. Serum inflammation biomarkers, oxidative stress markers, inflammatory cytokines and liver function enzymes were evaluated. Results: The results indicated that DHZ significantly ameliorated arthritis severity as shown by reduced arthritic score, thymus and spleen indexes, paw circumference, paw withdrawal threshold and latency as well as increased body weight gain. Furthermore, DHZ treatment persuasively reduced serum levels of alkaline phosphatase (ALP), aspartate transaminase (AST), alanine transaminase (ALT), rheumatoid factor (RF), C-reactive protein (CRP), tumor necrosis factor alpha (TNF-α), interleukin-1ß and 6 (IL-1ß and IL-6), malondialdehyde (MDA), vascular endothelial growth factor (VEGF) and transforming growth factor ß (TGF-ß). In addition, DHZ observably increased serum superoxide dismutase (SOD) and glutathione (GSH) levels in treated rats. Conclusion: These findings suggest that DHZ possesses anti-RA effect properties via modulating the inflammatory responses and oxidative stress.

Ameliorative potentials of the ethanolic extract from Lycium chinense leaf extract against diabetic cardiomyopathy. Insight into oxido-inflammatory and apoptosis modulation.

The prevalence of cardiovascular complications in diabetes has become one of the major cause of diabetes related morbidity/mortality. The onset and progression of diabetic cardiomyopathy (DCM) has been majorly linked to lipid alterations, oxidative stress, inflammation and apoptosis. This present study investigated the cardioprotective role of Lycium chinense leaf extract (LCME) in fructose/streptozotocin induced diabetic rats. Diabetic animals were orally gavaged with LCME (100 and 400 mg/kg) for five weeks. The results indicated that diabetic rats showed increased blood glucose concentration, serum cardiac function markers (troponin T, creatine kinase-MB, aspartate aminotransferase and lactate dehydrogenase) and lipid profile (triglycerides and cholesterol). In addition, the cardiac tissues of diabetic rats showed increased levels of nuclear factor-κB (NF-κB), tumor necrosis factor alpha (TNF-α), interleukin 1 beta (IL 1ß), interleukin 6 (IL-6), caspase-3 and malondialdehyde as well as significantly reduced activities of catalase, superoxide dismutase, reduced glutathione and glutathione peroxidase. LCME significantly ameliorated hyperglycemia and markedly decreased serum concentrations of troponin T, creatine kinase-MB, aspartate aminotransferase and lactate dehydrogenase, triglycerides and cholesterol. Furthermore, LCME notably suppressed cardiac oxido-inflammatory mediators and boosted cardiac antioxidant defense. Histopathologically, LCME restored cardiac structural alterations and also suppressed the immunohistochemical expression of collagen IV, smooth muscle alpha-actin (α-SMA) and p53, while Bcl2 expression was significantly increased. In conclusion, our result indicated that LCME protected against diabetic cardiomyopathy suppressing oxidative stress, inflammation, apoptosis and fibrosis.

Melatonin promotes sirtuin 1 expression and inhibits IRE1α-XBP1S-CHOP to reduce endoplasmic reticulum stress-mediated apoptosis in chondrocytes.

Osteoarthritis (OA) is the most common chronic disease characterized by a loss of chondrocytes and the degeneration of cartilage. Inflammation plays an important role in the pathogenesis and progression of OA via the activation of the endoplasmic reticulum (ER) stress signaling pathway. In this study, we stimulated human primary chondrocytes with lipopolysaccharide (LPS) to reduce cell viability and induce chondrocyte apoptosis. LPS-stimulated human primary chondrocytes induced ER stress and significantly upregulated the ER chaperone glucose-regulated protein 78 (GRP78) and increased the expression level of C/EBP-homologous protein (CHOP), a key mediator of ER stress--induced apoptosis. Interestingly, melatonin treatment attenuated ER stress-mediated chondrocyte apoptosis. Melatonin inhibited the expression of cleaved caspase-3, cleaved caspase-10, Bax, CHOP, GRP78, cleaved caspase-4, phospho-inositol-requiring enzyme 1α (P-IRE1α), and spliced X-box-binding protein 1 (XBP1S). In an anterior cruciate ligament transection mouse model of OA, melatonin (50 and 150 mg/kg) dose-dependently relieved joint cartilage degeneration and inhibitied of chondrocyte apoptosis. Immunohistochemical analysis indicated that melatonin could promote SIRT1 the expression and inhibit CHOP and cleaved caspase-3 expression in OA mice. In conclusion, our findings demonstrate for the first time that melatonin inhibits the IRE1α-XBP1S-CHOP signaling pathway by promoting the expression of SIRT1 in LPS-treated human chondrocytes and delaying OA progression in vivo.

SLAMF6 is associated with the susceptibility and severity of rheumatoid arthritis in the Chinese population.

OBJECTIVES: A recently published genome-wide association study identified six novel loci associated with rheumatoid arthritis (RA) in Korean population. We aimed to investigate whether these newly reported RA-risk loci are associated with RA in the Chinese population and to further characterize the functional role of the susceptible gene. METHODS: The susceptible variants of RA were genotyped in 600 RA patients and 800 healthy controls, including rs148363003 of SLAMF6, rs117605225 of CXCL13, rs360136 of SWAP70, rs111597524 of NFKBIA, rs194757 of ZFP36L1 and rs1547233 of LINC00158. Synovial tissues were collected from the knee joint of 50 RA patients and 40 controls without osteoarthritis for the gene expression analysis. Inter-group comparisons were performed with the Chi-square test for genotyping data or with Student's t-test for gene expression analysis. RESULT: For rs148363003 of SLAMF6, RA patients were observed to have a significantly lower frequency of genotype CC (4.5% vs. 0.9%, p = 0.004) as compared with the controls. The frequency of allele C was remarkably higher in the patients than in the controls (11.5% vs. 8.0%, p = 0.002), with an odds ratio of 1.49 (95% CI = 1.16-1.92). There was no significant difference between the patients and the controls regarding genotype or allele frequency of the other 5 variants. The mRNA expression of SLAMF6 was 1.6 folds higher in the RA patients than in the controls. Moreover, SLAMF6 expression was 1.5 folds higher in patients with genotype CC than in the patients with genotype TT. CONCLUSIONS: SLAMF6 was associated with both the susceptibility and severity of RA in the Chinese population. Moreover, rs148363003 could be a functional variant regulating the tissue expression of SLAMF6 in RA patients. It is advisable to conduct further functional analysis for a comprehensive knowledge on the contribution of this variant to the development of RA.

Identification of a novel microRNA-141-3p/Forkhead box C1/ß-catenin axis associated with rheumatoid arthritis synovial fibroblast function in vivo and in vitro.

Rationale: Rheumatoid arthritis (RA) is a prototype of inflammatory arthritis in which synovial fibroblasts (SFs) play key roles in cartilage and bone destruction through tumor-like proliferation, migration, invasion and inflammation. This study aimed to research forkhead box protein C1 (FoxC1) and microRNA (miR)-141-3p, which modulate pathological changes in the synovial membrane, to find possible strategies for treating RA. Methods: FoxC1, ß-catenin and miR-141-3p gene expression in synovial tissues and SFs was quantified by real-time PCR; FoxC1 and ß-catenin protein levels were evaluated by immunohistochemistry, immunofluorescence, and Western blotting. We transiently transfected human SFs with FoxC1 and ß-catenin overexpression and silencing vectors and assessed proliferation, migration, invasion and inflammation by cell function and enzyme-linked immunosorbent assays. We also assessed downstream signaling activation using immunofluorescence, real-time PCR and Western blotting. Double luciferase, coimmunoprecipitation and chromatin immunoprecipitation assays were used to verify miR-141-3p, FoxC1 and ß-catenin gene and protein combinations. Finally, the therapeutic effects of FoxC1 silencing and miR-141-3p overexpression were evaluated in type II collagen-induced arthritis (CIA) rats. Results: We found that FoxC1 expression was significantly upregulated in synovium and SFs in both RA patients and rats with collagen-induced arthritis (CIA). FoxC1 overexpression increased ß-catenin messenger RNA (mRNA) and protein levels and upregulated cyclin D1, c-Myc, fibronectin and matrix metalloproteinase 3 (MMP3) mRNA and protein expression in RA SFs (RASFs). In contrast, FoxC1 knockdown reduced ß-catenin mRNA and protein levels as well as cyclin D1, c-Myc, and fibronectin mRNA and protein levels in RASFs. Furthermore, altering FoxC1 expression did not significantly change GSK3ß and pGSK3ß levels. FoxC1 overexpression promoted proliferation, migration, invasion and proinflammatory cytokine (interleukin (IL)-1ß, IL-6, and tumor necrosis factor-α (TNF-α)) production and reduced anti-inflammatory cytokine (IL-10) levels in RASFs. FoxC1 bound to the ß-catenin promoter, and ß-catenin mediated the FoxC1-induced pathological changes. We also observed downregulated microRNA (miR)-141-3p expression in SFs from both RA patients and CIA rats and further found that miR-141-3p bound to the FoxC1 3'UTR and suppressed FoxC1 expression. Intra-ankle miR-141-3p agomir or FoxC1-specific siRNA injection hindered CIA development in rats. Conclusions: FoxC1 and miR-141-3p participate in RA pathogenesis by mediating inflammation and SF proliferation, migration, and invasion and thus could be novel targets for RA therapy as a nonimmunosuppressive approach.

[Ineffective treatment technique of convex epiphysiodesis for congenital scoliosis].

OBJECTIVE: To discuss the long-term outcome of convex epiphysiodesis in the treatment for congenital scoliosis (CS). METHODS: The clinical data of 22 patients with hemivertebral deformity undergoing convex epiphysiodesis from the October 1998 to Febuary 2008 were respectively analyzed. There were 12 males and 10 females. The whole spine anteroposterior radiographs were taken preoperatively, at 3-month postoperatively and at the final follow-up to measure the main curve and the compensatory curve. The progression rate was calculated for each patient. Observing the correlation between the progression rate and annual progression of the scoliosis and age, gender, hemivertebral number, hemivertebral position, preoperative main curve Cobb angle and compensatory curve Cobb angle, comparing different ages, genders, hemivertebral number and position, and preoperative main curve Cobb angle on the progression of postoperative curve. RESULTS: The mean Cobb angle of main curve changed from (40.5±9.8) ° before surgery to (39.5±11.1) ° at 3 months after surgery, which significantly increased to (46.8±13.9) ° in the final follow-up. Meanwhile the mean Cobb angle of compensatory curve was changed from (20.1±10.8) ° before surgery to (23.0±11.1) °, which significantly increased to (29.9±11.5) ° in the final follow-up. There were no significant differences in the Cobb angle of the main curve and the compensatory curve between postoperative 3 months and before operation (P>0.05). The difference between the final follow-up and the preoperative, postoperative 3 months was statistically significant (P<0.01). Twenty patients experienced progression of both main curve and compensatory curve, with a mean progression rate of (19.2±17.9)% for main curve and (39.6±37.0)% for compensatory curve. The annual progression volume was (1.5± 1.4) ° for main curve and (1.4±1.3) ° for compensatory curve. Three patients underwent lateral convex orthopedic internal fixation due to postoperative scoliosis progression. The curve progression was significantly correlated with age at the time of surgery and hemivertebral number. There was a significant correlation between the age of the operation, the main curve angle, the preoperative compensatory curve angle and the annual progression volume of the main curve (P<0.05). CONCLUSION: The convex epiphysiodesis technique cannot effectively prevent curve progression of CS patients in the long-term follow-up. It is not recommended to apply this technique to the treatment of patients with congenital hemivertebrae.

A novel variant near LSP1P3 is associated with knee osteoarthritis in the Chinese population.

OBJECTIVES: Previous genome-wide association study showing a novel variant near LSP1P3 was associated with knee osteoarthritis (KOA) in Caucasians. Replication study in different populations was essential to validate the association of novel susceptible genes with KOA. To our knowledge, there is a lack of study concerning the role of the LSP1P3 gene in Chinese KOA patients. We aimed to determine the association between the novel variant near LSP1P3 gene and the susceptibility of KOA in the Chinese population and to further investigate its relationship with the severity of KOA. METHODS: A total of 532 primary KOA patients who received treatment in our clinic center were included in the current study. Nine hundred twenty-seven age- and gender-matched healthy subjects were recruited as controls. The severity of KOA was graded according to the Kellgren-Lawrence (KL) grading system, with KL grade of 1 or 2 classified as mild KOA and KL grade of 3 or 4 classified as severe KOA. Three variants were genotyped using TaqMan SNP genotyping assay, including rs4867568 of LSP1P3 gene, rs143383 of GDF5, and rs1558902 of FTO. The differences in terms of genotype and allele distributions between the cases and the controls were analyzed by the chi-square test. RESULTS: There were 215 male and 317 female patients with a mean age of 58.1 ± 7.2 years. According to the KL score, 172 (32.3%) patients had mild KOA and 360 (67.7%) had severe KOA. There were remarkably lower frequencies of allele T of rs4867568 and allele C of rs143383 in the patients than in the controls (31.5% vs. 36.0%, p = 0.01 for rs4867568; 24.6% vs. 28.7%, p = 0.02 for rs143383), with an OR of 0.82 and 0.81, respectively. As for rs1558902, no significant difference regarding the frequency of allele and genotype was found between the patients and the controls. Patients with severe KOA had remarkably lower incidence of genotype TT of rs4867568 than patients with mild KOA (6.7% vs. 12.2%, p = 0.04). There was significantly higher frequency of allele T in patients with mild KOA than in those with severe KOA (36.3% vs. 29.2%, p = 0.02, OR = 0.72). CONCLUSIONS: The association of rs4867568 and rs143383 with KOA was successfully replicated in the Chinese Han population. Moreover, rs4867568 was found significantly associated with the severity of KOA. More studies are warranted to explore the functional role of rs4867568 in the development of KOA. Key Points • A novel variant near Lsp1p3 is associated with knee osteoarthritis. • Baseline characteristics of the subjects. • Comparison of the genotype and allele frequency of the Lsp1p3, GDF5, and FTO. • Association of the Lsp1p3, GDF5, and FTO with KOA severity.

Evodiamine attenuates adjuvant-induced arthritis in rats by inhibiting synovial inflammation and restoring the Th17/Treg balance.

OBJECTIVES: Evodiamine (Evo) possesses strong anti-inflammatory activity. In this study, we determine the antiarthritic effect of Evo. METHODS: Evo was administered to rats with adjuvant-induced arthritis (AA). We evaluated arthritis symptoms & histopathological changes and measured inflammatory cell infiltration, pro-inflammatory cytokine production and Th17 & Treg percentages in arthritic rats. KEY FINDINGS: Evo significantly improved the clinical signs of AA in rats, including decreases in paw swelling, the polyarthritis index and the number of swollen paw joints. Based on the histopathological analysis, Evo improved synovial inflammation and bone injury by inhibiting inflammatory cell infiltration, synoviocyte proliferation, pannus formation and cartilage erosion. Furthermore, the numbers of synovial CD3+ or CD68+ inflammatory cells were reduced, and the elevated levels of tumour necrosis factor-α, interleukin-1ß (IL-1ß) and IL-6 were restored to control levels by the Evo treatment. In addition, Evo therapy regulated the abnormal differentiation of Treg and Th17 cells, decreasing IL-17 production and increasing IL-10 levels. Finally, Evo inhibited Stat3 phosphorylation and induced Stat5 phosphorylation in rats with AA. CONCLUSIONS: Based on our results, Evo is a promising antiarthritic agent, potentially due to its inhibitory effect on synovial inflammation and regulatory effects on Treg and Th17 differentiation.

Forkhead box C1 promotes the pathology of osteoarthritis by upregulating ß-catenin in synovial fibroblasts.

Osteoarthritis (OA) is a joint disease characterized by articular cartilage degeneration, and no effective treatment is available. The OA classification has shifted from a cartilage-only disease to a whole-joint disease, and the synovial membrane plays an important role. Therefore, studies are needed to identify additional genes that regulate the pathological changes in the synovial membrane to develop a promising therapeutic strategy for OA. Here, we validated that the expression of forkhead box protein C1 (FoxC1) and ß-catenin was upregulated in OA synovial membranes and synovial fibroblasts (SFs). Gain- and loss-of-function studies revealed that FoxC1 overexpression promoted, whilst silencing inhibited OA synovial fibroblast (OASF) proliferation and pro-inflammatory cytokine [interleukin 6 (IL-6), interleukin 8 (IL-8) and tumour necrosis factor-α (TNF-α)] production. FoxC1 overexpression increased ß-catenin mRNA, total and nuclear protein expression in OASFs and upregulated a disintegrin and metalloproteinase with thrombospondin motif 5 (ADAMTS-5), fibronectin, matrix metalloproteinase 3 (MMP3) and matrix metalloproteinase 13 (MMP13) mRNA and total protein expression in OASFs. Conversely, FoxC1 knockdown reduced ß-catenin mRNA, total and nuclear protein expression in OASFs and reduced ADAMTS-5, fibronectin, MMP3 and MMP13 mRNA and total protein expression in OASFs. ß-catenin mediates FoxC1-induced pathological changes (proliferation, catabolic regulation and inflammation) in OASFs. MicroRNA-200a-3p (miR-200a-3p) binds to the 3'-UTR of FoxC1 and mediates FoxC1 expression. Intra-articular FoxC1-specific siRNA transfection hindered OA development in mice. Therefore, our results demonstrate the key role FoxC1 plays in vivo and in vitro in OA synovial pathology, possibly identifying a potential novel therapeutic target for OA.

Risk Factors of Impaired Pulmonary Function in Arthrogryposis Multiplex Congenital Patients With Concomitant Scoliosis: A Comparison With Adolescent Idiopathic Scoliosis.

STUDY DESIGN: A retrospective study. OBJECTIVE: To investigate the severity of pulmonary function impairment for arthrogryposis multiplex congenital (AMC) patients with concomitant scoliosis and to determine risk factors associated with the impaired pulmonary function in these patients. SUMMARY OF BACKGROUND DATA: AMC patients are generally believed to have impaired pulmonary function. However, the severity of respiratory morbidity and the associated risk factors have not been reported. METHODS: The pulmonary function tests data including the percentage predicted values of forced vital capacity (%FVC), forced expiratory volume in 1 second (%FEV1), and the ratio of FEV1 to FVC (%FEV1/FVC) were reviewed and compared for 48 AMC patients with secondary scoliosis and 48 patients with adolescent idiopathic scoliosis. The radiographic parameters of coronal and sagittal plane deformities and body mass index (BMI) were measured and correlated with impaired pulmonary function in these AMC patients. RESULTS: AMC patients with concomitant scoliosis had significant lower mean %FVC, %FEV1, and %FEV1/FVC than adolescent idiopathic scoliosis patients (48.8 vs. 70.3 for %FVC, P < 0.001; 45.3 vs. 69.7 for %FEV1, P < 0.001; 92.1 vs. 96.9 for %FEV1/FVC, P < 0.05, respectively). Seventy percent of AMC patients had severe pulmonary function impairment. A positive correlation was found between BMI and %FVC and %FEV1 (P < 0.01) and between hypokyphosis and %FVC and %FEV1 (P < 0.05). A negative correlation was found between coronal angle and pulmonary function (P < 0.05). Multiple regression analysis showed that all of the three variables were independent predictors associated with a reduced pulmonary function. With a combination of these variables, the multiple regression model could account for 38.7% of the variance in %FVC and 41.5% in %FEV1. CONCLUSION: There exists severe impairment of pulmonary function in AMC patients with concomitant scoliosis. The scoliotic curve, hypokyphosis, and BMI were independent risk factors associated with the pulmonary dysfunction in these patients. As the majority of the variability of impaired pulmonary function tests are still poorly understood, more risk factors await to be uncovered in future studies. LEVEL OF EVIDENCE: 4.

The Efficacy of Brace Treatment for Thoracolumbar Kyphosis in Patients With Achondroplasia.

STUDY DESIGN: A retrospective study. OBJECTIVES: The aim of this study was to evaluate the outcome of brace treatment in the correction of thoracolumbar kyphosis (TLK) for patients with achondroplasia and to determine the factors associated with bracing efficacy. SUMMARY OF BACKGROUND DATA: Brace treatment has been used to correct TLK in patients with achondroplasia. However, there was a paucity of knowledge concerning its effectiveness. METHODS: A total of 33 achondroplasic patients treated by bracing were included in this study. Radiographic parameters including TLK, lumbar lordosis, curve magnitude, apical vertebral translation (AVT), percentage of apical vertebral wedging, pelvic tilt (PT), and pelvic incidence were recorded for each patient at the visit. Comparison of these parameters between the initial visit and the final visit was performed using the Student t test. Factors associated with the correction of TLK were evaluated using the logistic regression analysis. RESULTS: The mean age at presentation was 27.5 ±â€Š13.4 months. The mean period of treatment was 32.2 ±â€Š15.7 months, and the mean period of follow-up was 25.7 ±â€Š11.3 months. At the initial visit, the mean value of TLK and the percentage of apical vertebral wedging were 41.7 ±â€Š15.4° and 61.4% ±â€Š16.2%, respectively. At the final visit, the TLK and apical vertebral wedging were remarkably reduced to 29.5 ±â€Š20.8° and 52.1% ±â€Š18.7%, respectively. The logistic regression analysis showed that initial TLK, AVT, percentage of apical vertebral wedging, and PT were independent factors associated with the correction of TLK. CONCLUSION: Brace treatment can effectively correct TLK and restore the morphology of apical vertebral body for patients with achondroplasia. Large TLK, severe apical vertebral wedging, presence of AVT, and low PT may be indicative of an unfavorable outcome, which should be taken into account at the initiation of bracing. LEVEL OF EVIDENCE: 4.

Common Variant of POC5 Is Associated With the Susceptibility of Adolescent Idiopathic Scoliosis.

STUDY DESIGN: A case-control study. OBJECTIVE: To validate the relationship between POC5 and adolescent idiopathic scoliosis (AIS) in the Chinese patients and to further investigate the functional role of POC5. SUMMARY OF BACKGROUND DATA: Three rare functional variants in the POC5 were recently reported to be strongly associated with the disease in a large family with multiple members affected with idiopathic scoliosis. To our knowledge, the association between the mutations of POC5 and AIS remains undetermined in the Chinese population. METHODS: Single nucleotide variants c.1336G>A, c.1286C>T, and c.1363G>C of POC5 were genotyped in 2432 patients with AIS and 2292 healthy controls using multiple ligase detection reactions. Common variants covering POC5 gene were genotyped in 1446 patients and 2080 controls. The mRNA expression of POC5 was determined in the paraspinal muscles collected from 98 patients and 28 controls. The Student t test was used to compare mRNA expression level between the patients and the controls. In addition, the POC5 expression was compared among different genotypes of the remarkably associated single nucleotide polymorphism (SNP) with analysis of variance test. RESULTS: There was no case of mutation for the three reported variants of POC5. SNP rs6892146 was observed to have significantly different distribution of minor allele frequency in the two group (0.485 vs. 0.446, P = 0.004). The mRNA expression of POC5 was 1.5-fold higher in patients than in the controls (0.00012 ±â€Š0.00009 vs. 0.00008 ±â€Š0.00006, P = 0.02). Patients with genotype GG have a significantly increased expression of POC5 than those with CC (0.00014 ±â€Š0.00007 vs. 0.00009 ±â€Š0.00007, P = 0.03). CONCLUSION: Common variant rs6892146 of POC5 is associated with the development of AIS in the Chinese population. Targeted regional sequencing of POC5 may help identify novel mutations associated with AIS. LEVEL OF EVIDENCE: 4.

Kirschner wire is more effective than the nerve root retractor in treating patients with disc herniation.

OBJECTIVE: To investigate the value of using Kirschner wire in the intraspinal procedures. METHODS: From May 2011 to October 2013, a total of 46 patients with a single-level lumbar disc herniation with concomitant lumbar instability were randomly assigned to two groups at the time of admission. Group 1 had 23 patients who underwent posterior lumbar fusion using a nerve root retractor to drag nerve root, and Group 2 also had 23 patients who underwent the same operation by fixing Kirschner wires in the vertebral body to drag nerve root. All of these patients were assessed with visual analog scales (VAS) and Japanese Orthopaedic Association (JOA) scores before surgery, and were followed up at the time points of one week (VAS score only), three, six, and twelve months respectively after surgery. The actual retraction time of the nerve root of each patient was also recorded during the operation. RESULTS: The differences in VAS and JOA scores were not significant between Group 1 and 2 before surgery. However, these scores showed significant improvement in Group 2 at one week and three months after surgery compared with those in Group 1. At six and twelve month follow-up time points, no significant difference was observed between these two groups. However, the retraction time of the nerve root of the patients in Group 2 was significantly shorter than in Group 1. CONCLUSIONS: Using Kirschner wires instead of the nerve root retractor to pull nerve root in the patients with a single-level lumbar disc herniation accompanied by the lumbar instability is more effective in reducing the dragging damage of the nerve root at early phases after surgery and in shortening retraction time.

Complications after selective embolization in the bilateral internal iliac arteries and the median sacral artery with gelfoam particles in dogs.

OBJECTIVE: The study aimed to observe the complications after the bilateral internal iliac arteries and the median sacral artery embolization by different severity and combinations of gelfoam particles. METHODS: Sixteen healthy adult dogs were randomly divided into five groups. Under the monitoring of digital subtraction angiography (DSA), gelfoam particles with diameter of 50-150 µm were applied. In group A, embolization was performed up to the trunk of bilateral internal iliac arteries and the median sacral artery; in group B, embolization was up to the trunk of bilateral internal iliac arteries; in group C, embolization was up to the first branch of bilateral internal iliac arteries and the median sacral artery; in group D, embolization was up to the trunk of unilateral internal iliac artery and the median sacral artery; in group E embolization was performed up to the trunk of unilateral internal iliac artery. RESULTS: Seven dogs died within 48 hours after embolization. In the dead animals of groups A, C and D, there were rectum necrosis and lamellar obfuscation and hemorrhage edema in bladder. In the histological examination, there are rectum and bladder cell dissociation, inflammatory cell infiltration and epithelial cell ablating in the dead animals. The embolization mainly presented in arterioles with a diameter of 100-200 µm. CONCLUSION: When gelfoam particles of 50-150 µm in diameter were applied for embolization in the internal iliac artery and median sacral artery, at least unilateral internal iliac artery should be preserved when embolization is performed in the proximal artery and the trunk.