6-formylindolo[3, 2-b]carbazole alters gut microbiota and prevents the progression of ankylosing spondylitis in mice.

Ankylosing spondylitis (AS), is known as a chronic inflammatory autoimmune disease, there is evidence to suggest that gut microbiota disorders may be related to the occurrence and development of AS. Studies have shown that 6-formylindolo[3, 2-b]carbazole (FICZ) has the ability to modulate intestinal homeostasis and inhibit inflammatory responses. The purpose of this work is to evaluate the protective role of FICZ in treating AS and elucidate potential mechanisms. FICZ was administered to the proteoglycan (PG)-induced AS mice for 7 consecutive weeks. The effects of FICZ on AS mice were evaluated by the disease severity, intestinal histopathology, proinflammatory cytokine levels, and intestinal mucosal barrier function. The gut microbiota compositions were profiled through 16S rDNA high-throughput sequencing. We found that FICZ significantly reduced the severity of AS and resulted in the downregulating of TNF-α and IL-17A inflammatory cytokines. Moreover, FICZ ameliorated pathological changes in the ileal and improved intestinal mucosal barrier function. Furthermore, FICZ altered the composition of the gut microbiota by increasing the Bacteroidetes/Firmicutes phylum ratio and enriched the genes related to "glycan biosynthesis and metabolism", thus reversing the process of AS. In conclusion, FICZ suppressed the progression of AS and altered gut microbiota in AS mice, which provided new insight into AS therapy strategy.

Deep Learning-Based Automated Magnetic Resonance Image Segmentation of the Lumbar Structure and Its Adjacent Structures at the L4/5 Level.

(1) Background: This study aims to develop a deep learning model based on a 3D Deeplab V3+ network to automatically segment multiple structures from magnetic resonance (MR) images at the L4/5 level. (2) Methods: After data preprocessing, the modified 3D Deeplab V3+ network of the deep learning model was used for the automatic segmentation of multiple structures from MR images at the L4/5 level. We performed five-fold cross-validation to evaluate the performance of the deep learning model. Subsequently, the Dice Similarity Coefficient (DSC), precision, and recall were also used to assess the deep learning model's performance. Pearson's correlation coefficient analysis and the Wilcoxon signed-rank test were employed to compare the morphometric measurements of 3D reconstruction models generated by manual and automatic segmentation. (3) Results: The deep learning model obtained an overall average DSC of 0.886, an average precision of 0.899, and an average recall of 0.881 on the test sets. Furthermore, all morphometry-related measurements of 3D reconstruction models revealed no significant difference between ground truth and automatic segmentation. Strong linear relationships and correlations were also obtained in the morphometry-related measurements of 3D reconstruction models between ground truth and automated segmentation. (4) Conclusions: We found it feasible to perform automated segmentation of multiple structures from MR images, which would facilitate lumbar surgical evaluation by establishing 3D reconstruction models at the L4/5 level.

Automated Magnetic Resonance Image Segmentation of Spinal Structures at the L4-5 Level with Deep Learning: 3D Reconstruction of Lumbar Intervertebral Foramen.

OBJECTIVE: 3D reconstruction of lumbar intervertebral foramen (LIVF) has been beneficial in evaluating surgical trajectory. Still, the current methods of reconstructing the 3D LIVF model are mainly based on manual segmentation, which is laborious and time-consuming. This study aims to explore the feasibility of automatically segmenting lumbar spinal structures and increasing the speed and accuracy of 3D lumbar intervertebral foramen (LIVF) reconstruction on magnetic resonance image (MRI) at the L4-5 level. METHODS: A total of 100 participants (mean age: 42.2 ± 14.0 years; 52 males and 48 females; mean body mass index, 22.7 ± 3.2 kg/m2 ), were enrolled in this prospective study between March and July 2020. All participants were scanned on L4-5 level with a 3T MR unit using 3D T2-weighted sampling perfection with application-optimized contrast with various flip-angle evolutions (SPACE) sequences. The lumbar spine's vertebra bone structures (VBS) and intervertebral discs (IVD) were manually segmented by skilled surgeons according to their anatomical outlines from MRI. Then all manual segmentation were saved and used for training. An automated segmentation method based on a 3D U-shaped architecture network (3D-UNet) was introduced for the automated segmentation of lumbar spinal structures. A number of quantitative metrics, including dice similarity coefficient (DSC), precision, and recall, were used to evaluate the performance of the automated segmentation method on MRI. Wilcoxon signed-rank test was applied to compare morphometric parameters, including foraminal area, height and width of 3D LIVF models between automatic and manual segmentation. The intra-class correlation coefficient was used to assess the test-retest reliability and inter-observer reliability of multiple measurements for these morphometric parameters of 3D LIVF models. RESULTS: The automatic segmentation performance of all spinal structures (VBS and IVD) was found to be 0.918 (healthy levels: 0.922; unhealthy levels: 0.916) for the mean DSC, 0.922 (healthy levels: 0.927; unhealthy levels: 0.920) for the mean precision, and 0.917 (healthy levels: 0.918; unhealthy levels: 0.917) for the mean recall in the test dataset. It took approximately 2.5 s to achieve each automated segmentation, far less than the 240 min for manual segmentation. Furthermore, no significant differences were observed in the foraminal area, height and width of the 3D LIVF models between manual and automatic segmentation images (P > 0.05). CONCLUSION: A method of automated MRI segmentation based on deep learning algorithms was capable of rapidly generating accurate segmentation of spinal structures and can be used to construct 3D LIVF models from MRI at the L4-5 level.

Indole-3-Acetic Acid Alters Intestinal Microbiota and Alleviates Ankylosing Spondylitis in Mice.

Ankylosing spondylitis (AS) is a systemic, chronic, and inflammatory autoimmune disease associated with the disorder of intestinal microbiota. Unfortunately, effective therapies for AS are lacking. Recent evidence has indicated that indole-3-acetic acid (IAA), an important microbial tryptophan metabolite, can modulate intestinal homeostasis and suppress inflammatory responses. However, reports have not examined the in vivo protective effects of IAA against AS. In this study, we investigated the protective effects and underlying mechanisms through which IAA acts against AS. We constructed a proteoglycan (PG)-induced AS mouse model and administered IAA (50 mg/kg body weight) by intraperitoneal injection daily for 4 weeks. The effects of IAA on AS mice were evaluated by examining disease severity, intestinal barrier function, aryl hydrocarbon receptor (AhR) pathway, T-helper 17 (Th17)/T regulatory (Treg) balance, and inflammatory cytokine levels. The intestinal microbiota compositions were profiled through whole-genome sequencing. We observed that IAA decreased the incidence and severity of AS in mice, inhibited the production of pro-inflammatory cytokines (tumor necrosis factor α [TNF-α], interleukin [IL]-6, IL-17A, and IL-23), promoted the production of the anti-inflammatory cytokine IL-10, and reduced the ratios of pro-/anti- inflammatory cytokines. IAA ameliorated pathological changes in the ileum and improved intestinal mucosal barrier function. IAA also activated the AhR pathway, upregulated the transcription factor forehead box protein P3 (FoxP3) and increased Treg cells, and downregulated the transcription factors retinoic acid receptor-related orphan receptor gamma t (RORγt) and signal transducer and activator of transcription 3 (STAT3) and decreased Th17 cells. Furthermore, IAA altered the composition of the intestinal microbiota composition by increasing Bacteroides and decreasing Proteobacteria and Firmicutes, in addition to increasing the abundances of Bifidobacterium pseudolongum and Mucispirillum schaedleri. In conclusion, IAA exerted several protective effects against PG-induced AS in mice, which was mediated by the restoration of balance among the intestinal microbial community, activating the AhR pathway, and inhibiting inflammation. IAA might represent a novel therapeutic approach for AS.

Computerized Characterization of Spinal Structures on MRI and Clinical Significance of 3D Reconstruction of Lumbosacral Intervertebral Foramen.

BACKGROUND: Segmentation of spinal structures is important in medical imaging analysis, which facilitates surgeons to plan a preoperative trajectory for the transforaminal approach. However, manual segmentation of spinal structures is time-consuming, and studies have not explored automatic segmentation of spinal structures at the L5/S1 level. OBJECTIVES: This study sought to develop a new method based on a deep learning algorithm for automatic segmentation of spinal structures. The resulting algorithm may be used to rapidly generate a precise 3D lumbosacral intervertebral foramen model to assist physicians in planning an ideal trajectory in L5/S1 lumbar transforaminal radiofrequency ablation (LTRFA). STUDY DESIGN: This was an observational study for developing a new technique on spinal structures segmentation. STUDY SITE: The study was carried out at the department of radiology and spine surgery at our hospital. METHODS: A total of 100 L5/S1 level data samples from 100 study patients were used in this study. Masks of vertebral bone structures (VBSs) and intervertebral discs (IVDs) for all data samples were segmented manually by a skilled surgeon and served as the "ground truth." After data preprocessing, a 3D-UNet model based on deep learning was used for automated segmentation of lumbar spine structures at L5/S1 level magnetic resonance imaging (MRI). Segmentation performances and morphometric measurement were used for 3D lumbosacral intervertebral foramen (LIVF) reconstruction  generated by either manual segmentation and automatic segmentation. RESULTS: The 3D-UNet model showed high performance in automatic segmentation of lumbar spinal structures (VBSs and IVDs). The corresponding mean Dice similarity coefficient (DSC) of 5-fold cross-validation scores for L5 vertebrae, IVDs, S1 vertebrae, and all L5/S1 level spinal structures were 93.46 ± 2.93%, 90.39 ± 6.22%, 93.32 ± 1.51%, and 92.39 ± 2.82%, respectively. Notably, the analysis showed no associated difference in morphometric measurements between the manual and automatic segmentation at the L5/S1 level. LIMITATIONS: Semantic segmentation of multiple spinal structures (such as VBSs, IVDs, blood vessels, muscles, and ligaments) was simultaneously not integrated into the deep-learning method in this study. In addition, large clinical experiments are needed to evaluate the clinical efficacy of the model. CONCLUSION: The 3D-UNet model developed in this study based on deep learning can effectively and simultaneously segment VBSs and IVDs at L5/S1 level formMR images, thereby enabling rapid and accurate 3D reconstruction of LIVF models. The method can be used to segment VBSs and IVDs of spinal structures on MR images within near-human expert performance; therefore, it is reliable for reconstructing LIVF for L5/S1 LTRFA.

Identification of aberrantly methylated differentially expressed genes targeted by differentially expressed miRNA in osteosarcoma.

BACKGROUND: Osteosarcoma (OS) is the most common primary bone tumors diagnosed in children and adolescents. Recent studies have shown a prognostic role of DNA methylation in various cancers, including OS. The aim of this study was to identify the aberrantly methylated genes that are prognostically relevant in OS. METHODS: The differentially expressed mRNAs, miRNAs and methylated genes (DEGs, DEMs and DMGs respectively) were screened from various GEO databases, and the potential target genes of the DEMs were predicted by the RNA22 program. The protein-protein interaction (PPI) networks were constructed using the STRING database and visualized by Cytoscape software. The functional enrichment and survival analyses of the screened genes was performed using the R software. RESULTS: Forty-seven downregulated hypermethylated genes and three upregulated hypomethylated genes were identified that were enriched in cell activation, migration and proliferation functions, and were involved in cancer-related pathways like JAK-STAT and PI3K-AKT. Eight downregulated hypermethylated tumor suppressor genes (TSGs) were identified among the screened genes based on the TSGene database. These hub genes are likely involved in OS genesis, progression and metastasis, and are potential prognostic biomarkers and therapeutic targets. CONCLUSIONS: TSGs including PYCARD, STAT5A, CXCL12 and CXCL14 were aberrantly methylated in OS, and are potential prognostic biomarkers and therapeutic targets. Our findings provide new insights into the role of methylation in OS progression.

Anti-TNF-α therapy alters the gut microbiota in proteoglycan-induced ankylosing spondylitis in mice.

Ankylosing spondylitis is a chronic, progressive disease, and its treatment is relevant to the gut microbiota. Anti-tumor necrosis factor-alpha (anti-TNF-α) therapy alters the gut microbiota in many diseases, including inflammatory bowel disease. However, little is known about the effect of TNF-α blocker treatment on the gut microbiota in ankylosing spondylitis. Herein, the effect of a TNF-α blocker on the gut microbiota in proteoglycan-induced arthritis was investigated. Proteoglycan-induced mice were treated with an rhTNFR:Fc solution of etanercept (5 µg/g) for 4 weeks. rhTNFR:Fc treatment attenuated the arthritis incidence and severity of arthritis in the proteoglycan-induced mice and decreased inflammation in the ankle joints and ameliorated ileal tissue destruction. Moreover, high gut permeability occurred, and zonula occludens-1 and occludin protein levels were reduced in proteoglycan-induced mice. These levels were significantly restored by the administration of rhTNFR:Fc. The serum TNF-α and IL-17 levels were also decreased. In addition, flora analysis via 16S rDNA high-throughput sequencing revealed that rhTNFR:Fc treatment restored the gut microbiota composition to a composition similar to that in control mice. In conclusion, anti-TNF-α therapy attenuated proteoglycan-induced arthritis progression and modulated the gut microbiota and intestinal barrier function. These results provide new insights for anti-TNF-α therapy strategies via regulating the gut microbiota in ankylosing spondylitis.

Clinical Anatomy and Possible Clinical Significance of the Intervertebral Vein in the Lumbar Intervertebral Foramina.

BACKGROUND: The detailed structure of the lumbar intervertebral foramina has been well-studied. Nevertheless, detailed descriptions of branches of the intervertebral vein (IV) through the lumbar intervertebral foramina are lacking. OBJECTIVES: This study aimed to provide an anatomical basis for invasive treatment targeting the branches of the IV using an approach through the lumbar intervertebral foramina, particularly for the purposes of transforaminal epidural steroid injection. STUDY DESIGN: This research involved a dissection-based study of 10 embalmed human cadavers. SETTING: The research took place at The Third Affiliated Hospital of Southern Medical University. METHODS: One hundred lumbar intervertebral foramina from 10 embalmed cadavers were studied. Branches of the IV in the intervertebral foramina were observed. The length and diameter of the veins were measured using a Vernier caliper. RESULTS: At a rate of 100%, branches of the IV were observed in the 100 lumbar foramina examined in our study. The following 4 types of branches of the IV were routinely found: Type I in 27 (27%) of the IV foramina, in which a superior branch of the IV ran along the inferior margin of the vertebral pedicle; Type II in 18 (18%) of the intervertebral foramina, in which an inferior branch of the IV ran along the superior margin of the inferior vertebral pedicle; Type III in 41 (41%) of the intervertebral foramina, in which the IV was divided into a superior and inferior branch; and Type IV in 14 (14%) of the intervertebral foramina, in which the IV was divided into 2 superior branches and an inferior branch. LIMITATIONS: The greatest weakness of this study is that it lacks actual clinical verification. Future clinical trials are expected to contribute more objective data concerning the IV branches. Due to the relative changes in vascular position during dissection, the relevant data warrant improvement. CONCLUSIONS: The lumbar IVs are an important part of the anatomical structure of the intervertebral foramina. Adequate knowledge of the IV may be of clinical importance to physicians performing transforaminal epidural steroid injection. KEY WORDS: Clinical anatomy, intervertebral veins, lumbar vertebra, Kambin's triangle, safe triangle, intervertebral foramina, vertebral venous system, inadvertent injection, transforaminal epidural steroid injection.

Rifaximin Alters Intestinal Microbiota and Prevents Progression of Ankylosing Spondylitis in Mice.

Recently, accumulating evidence has suggested that gut microbiota may be involved in the occurrence and development of ankylosing spondylitis (AS). It has been suggested that rifaximin have the ability to modulate the gut bacterial communities, prevent inflammatory response, and modulate gut barrier function. The goal of this work is to evaluate the protective effects of rifaximin in fighting AS and to elucidate the potential underlying mechanism. Rifaximin were administered to the proteoglycan (PG)-induced AS mice for 4 consecutive weeks. The disease severity was measured with the clinical and histological of arthritis and spondylitis. Intestinal histopathological, pro-inflammatory cytokine levels and the intestinal mucosal barrier were evaluated. Then, western blot was performed to explore the toll-like receptor 4 (TLR-4) signal transducer and NF-κB expression. Stool samples were collected to analyze the differences in the gut microbiota via next-generation sequencing of 16S rDNA. We found that rifaximin significantly reduced the severity of AS and resulted in down-regulation of inflammatory factors, such as TNF-α, IL-6, IL-17A, and IL-23. Meanwhile, rifaximin prevented ileum histological alterations, restored intestinal barrier function and inhibited TLR-4/NF-κB signaling pathway activation. Rifaximin also changed the gut microbiota composition with increased Bacteroidetes/Firmicutes phylum ratio, as well as selectively promoting some probiotic populations, including Lactobacillales. Our results suggest that rifaximin suppressed progression of AS and regulated gut microbiota in AS mice. Rifaximin might be useful as a novel treatment for AS.

Long noncoding RNA DNAJC3-AS1 promotes osteosarcoma progression via its sense-cognate gene DNAJC3.

Long noncoding RNAs have been proved to play essential roles in tumor development and progression. In this study, we focused on DNAJC3-AS1 and investigated its biological function and clinical significance in osteosarcoma. We detected the expression of DNAJC3-AS1 in 30 pairs of matched osteosarcoma and adjacent nontumorous specimens and osteosarcoma cell lines and analyzed association between DNAJC3-AS1 levels and clinicopathological factors. We found that DNAJC3-AS1 expression was up-regulated in osteosarcoma. High level of DNAJC3-AS1 was correlated with high differentiated degree (P = 0.018) and advanced Enneking stage (P = 0.016). Mechanistically, DNAJC3-AS1 enhanced cell proliferation, migration, and invasion in vitro and in vivo and reduced sensitivity of osteosarcoma to cisplatin. These effects of DNAJC3-AS1 were reversed by down-regulation of its sense-cognate gene DNAJC3. Thus, DNAJC3-AS1 promotes osteosarcoma development and progression by regulating DNAJC3 and might be a biomarker and therapeutic target for osteosarcoma.

NVP-BEZ235 synergizes cisplatin sensitivity in osteosarcoma.

Osteosarcoma(OS) remains a major health concern in childhood and adolescence, although cisplatin is one of the gold standard chemotherapeutic drugs in the treatment of OS, chemoresistant to cisplatin is common. Phosphoinositide 3-kinase (PI3K)-Akt-mammalian target of rapamycin inhibitor (mTOR) pathway and autophagy regulates chemosensitivity incancer cells. In this study, we hypothesized that NVP-BEZ235, a dual inhibitor of PI3K/mTOR, could synergize cisplatin sensitivity in OS. In vitro, NVP-BEZ235 plus cisplatinexerted a synergistic effect on cell proliferation inhibition and apoptosis induction. Cisplatin could activate PI3K-Akt-mTOR pathway activity in early times, whereas, NVP-BEZ235 could inhibit PI3K-Akt -mTOR pathway activity all the times alone or combined with cisplatin. What's more, NVP-BEZ235 could switch function of autophagy induced by cisplatin to synergize cisplatin sensitivity. In vivo, pronounced decrease in tumor cell proliferation and increase in apoptosisin combination-treated mouse xenograft models compared with cisplatin or NVP-BEZ235 treated models. All these results suggest NVP-BEZ235 could synergize cisplatin sensitivity in OS, combination of NVP-BEZ235 with cisplatin could represent a novel therapeutic strategy for treatment of OS.

Glucan phosphate inhibits HMGB-1 release from rat myocardial H9C2 cells in sepsis via TLR4/NF-кB signal pathway.

PURPOSE: The effect of glucan phosphate (GP) on the release of HMGB-1 from rat myocardial cells (H9C2) during lipopolysaccharide-induced sepsis, and the underlying mechanisms, were investigated. METHODS: H9C2 cells were divided into three groups: normal; lipopolysaccharide (LPS) (1 mg/ml LPS); and, LPS+GP (2 mg/ml GP). Western blot was used to determine toll-like receptor 4 (TLR4) levels, and electrophoretic mobility-shift assays (EMSA) was used to determine nuclear factor-кB (NF-кB) activity 3, 6 and 9 h after treatment. HMGB-1 mRNA levels in cultured cells were determined by real-time PCR and supernatant HMGB-1 protein levels were evaluated by ELISA at 12, 24, 36 and 48 h after treatment. Following the transfection of H9C2 cells with Ad5-IкBα, which inhibits NF-кB activity, TLR4, NF-кB and HMGB-1 levels were determined. RESULTS: Intracellular TLR4 levels and NF-кB activity in LPS and LPS+GP groups increased 3-9 h after stimulation, but the increased levels of TLR4 and elevated activity of NF-кB were significantly lower in the LPS+GP group vs. the LPS group. HMGB-1 mRNA levels in both LPS and LPS+GP groups, increased gradually from 24 h after stimulation, but the increase was more obvious in the LPS group vs. the LPS+GP group. Supernatant HMGB-1 levels in the LPS and LPS+GP groups increased gradually from 9 h after stimulation, and also increased markedly in the LPS group. After the inhibition of NF-кB activity, LPS-induced HMGB-1 release decreased significantly (p.

Treatment of liver cancer of middle and advanced stages using ultrasound-guided percutaneous ethanol injection combined with radiofrequency ablation: A clinical analysis.

Liver cancer is a malignancy of the digestive system and has a high morbidity and mortality rate. Local intervention has become a viable option in identifying liver treatment. The aim of the present study was to analyze the clinical effects of treating liver cancer in middle and advanced stages using ultrasound-guided percutaneous ethanol injection (PEI) in tumors combined with radiofrequency ablation (RFA). A total of 100 patients with stage III-IV liver cancers were selected to participate in the study. Patients were divided into groups. In group A, treatment was initiated with PEI and after 1-2 weeks RFA was applied while in group B treatment was initiated with RFA and after 1-2 weeks PEI was applied. Patients in group C received PEI and RFA simultaneously. The clinical effects in the 3 groups were compared after 6-month follow ups. The volume of tumor ablation necrosis in group A was significantly greater than that in the groups B and C, while the size was significantly smaller compared to groups B and C after ablation. For group A, the complete ablation rate was significantly higher than that in groups B and C, and the differences were statistically significant (P<0.05). Liver damage indices, including raising levels of glutamic-pyruvic transaminase and total bilirubin, were significantly decreased in group A (P<0.05). The survival rate in group A was also significantly higher than in groups B and C (P<0.05). In conclusion, for patients with liver cancer in middle and advanced stages, the treatment method using PEI followed by RFA was more beneficial in terms of improving the tumor ablation rate, alleviating liver damages and increasing survival rates.