Association between Mean Corpuscular Hemoglobin and Platelet Distribution Width Ratio and Knee Osteoarthritis Severity.

BACKGROUND: The goal was to simply and efficiently predict the indicators of disease severity in knee osteoarthritis (KOA) patients. METHODS: One hundred eighty-four patients with KOA and 126 healthy subjects were included. WOMAC (Western Ontario and McMaster Universities Osteoarthritis Index) was used as a reference index for disease severity in KOA patients, in which WOMAC < 80 was classified as mild and WOMAC ≥ 80 as moderate and severe. Blood routine parameters of the KOA and the healthy groups were analyzed by the Mann Whitney U test. Receiver operating characteristic curves were used to analyze the sensitivity and specificity of mean corpuscular hemoglobin and platelet distribution width ratio (MPR) and monocyte and hemoglobin ratio (MHR) indicators. The correlation between MPR and MHR and disease severity of KOA was determined by bivariate regression analysis. Independent predictors of disease severity in patients with KOA were assessed by multivariate regression analysis. RESULTS: MPR, MHR, and WOMAC were significantly higher in the KOA group. The ROC curve indicated that the cutoff values of MPR and MHR were 2.09 and 0.0030, respectively, with sensitivity of 86.4% and 68.5% and specificity of 99.2% and 79.4%. Bivariate regression analysis found that MPR was better correlated with disease severity than MHR. The results of multivariate regression analysis demonstrated that the MPR values of moderate and severe patients were more than 19 times that of mild patients, and the OR values were 21.695 and 19.558, respectively. CONCLUSIONS: MPR and MHR demonstrated a good correlation with disease severity in patients with KOA. MPR is a potential independent predictor of disease severity in patients with KOA.

Biomechanical effects of an oblique lumbar interbody fusion combined with posterior augmentation: a finite element analysis.

BACKGROUND: Oblique lateral interbody fusion (OLIF) is widely used to treat lumbar degenerative disc disease. This study aimed to evaluate the biomechanical stability of OLIF, OLIF including posterior pedicle screw and rod (PSR), and OLIF including cortical screw and rod (CSR) instrumentation through finite element analysis. METHODS: A complete L2-L5 finite element model of the lumbar spine was constructed. Surgical models of OLIF, such as stand-alone, OLIF combined with PSR, and OLIF combined with CSR were created in the L3-L4 surgical segments. Range of motion (ROM), end plate stress, and internal fixation peak stress were compared between different models under the same loading conditions. RESULTS: Compared to the intact model, ROM was reduced in the OLIF model under all loading conditions. The surgical models in order of increasing ROM were PSR, CSR, and stand-alone; however, the difference in ROM between BPS and CSR was less than 0.4° and was not significant under any loading conditions. The stand-alone model had the highest stress on the superior L4 vertebral body endplate under all loading conditions, whereas the end plate stress was relatively low in the BPS and CSR models. The CSR model had the highest internal fixation stress, concentrated primarily at the end of the screw. CONCLUSIONS: OLIF alone significantly reduces ROM but does not provide sufficient stability. Addition of posterior PSR or CSR internal fixation instrumentation to OLIF surgery can significantly improve biomechanical stability of the segment undergoing surgery.

Surgical safe zones for oblique lumbar interbody fusion of L1-5: A cadaveric study.

To evaluate the operating range and morphology of the surgical safe zone for oblique lumbar interbody fusion (OLIF). Twenty embalmed full-torso cadaveric specimens were dissected. The oblique corridor and the distance between adjacent lumbar arteries were measured in a static state and with psoas major retraction. The morphology and size of the safe zone for OLIF and the location of the lumbar sympathetic trunk were also recorded. The oblique corridor of the L1-L5 segments was significantly greater in the retracted state than in the static state (p < 0.05). With psoas major retraction, the distances between adjacent lumbar arteries at L1-4 were significantly greater (p < 0.05) than those in the static state. The lumbar sympathetic trunk is just located in the safe zone and travels downward adjacent to the psoas major. The shape of the safe zone for OLIF was approximately an oblique upward parallelogram at L1/2 and L2/3, an isosceles trapezoid at L3/4, and an irregular quadrangle or triangle at L4/5. The safe zone for OLIF at L1/2, L2/3, and L3/4 was significantly larger during retraction than in the static state (p < 0.05). On the lateral side of the lumbar spine there is a natural surgical safe zone for OLIF, which can provide a sufficient operating space. The safe zone has a certain morphological pattern in L1-5 segments and psoas major retraction can significantly enlarge it.

Absence of VEGFR-1/Flt-1 signaling pathway in mice results in insensitivity to discogenic low back pain in an established disc injury mouse model.

Although degenerative disc disease (DDD) and related low back pain (LBP) are growing public health problems, the underlying disease mechanisms remain unclear. An increase in the vascular endothelial growth factor (VEGF) levels in DDD has been reported. This study aimed to examine the role of VEGF receptors (VEGFRs) in DDD, using a mouse model of DDD. Progressive DDD was induced by anterior stabbing of lumbar intervertebral discs in wild type (WT) and VEGFR-1 tyrosine-kinase deficient mice (vegfr-1TK-/- ). Pain assessments were performed weekly for 12 weeks. Histological and immunohistochemical assessments were made for discs, dorsal root ganglions, and spinal cord. Both vegfr-1TK-/- and WT mice presented with similar pathological changes in discs with an increased expression of inflammatory cytokines and matrix-degrading enzymes. Despite the similar pathological patterns, vegfr-1TK-/- mice showed insensitivity to pain compared with WT mice. This insensitivity to discogenic pain was related to lower levels of pain factors in the discs and peripheral sensory neurons and lower spinal glial activation in the vegfr-1TK- /- mice than in the WT mice. Exogenous stimulation of bovine disc cells with VEGF increased inflammatory and cartilage degrading enzyme. Silencing vegfr-1 by small-interfering-RNA decreased VEGF-induced expression of pain markers, while silencing vegfr-2 decreased VEGF-induced expression of inflammatory and metabolic markers without changing pain markers. This suggests the involvement of VEGFR-1 signaling specifically in pain transmission. Collectively, our results indicate that the VEGF signaling is involved in DDD. Particularly, VEGFR-1 is critical for discogenic LBP transmission independent of the degree of disc pathology.

Development and validation of a pre-scoring system for nonspecific low back pain among general population in Guangzhou: a cross-sectional study.

BACKGROUND: Nonspecific Low Back Pain (NLBP) is a common disease with a low cure rate and significant impact on the population. This study aimed to develop and validate a pre-scoring system for identifying the risk of suffering from NLBP among the general population in Guangzhou. METHODS: A total of 1439 eligible subjects were surveyed in Guangzhou by stratified random sampling and was divided randomly into the development dataset (69.6%) and validation dataset (30.4%) subsequently. Based on the development dataset, potential associated factors (average exercise times weekly, the intensity of daily work, etc.) with NLBP were tested by the sequential logistic regression, and a pre-scoring system was formulated with Sullivan's method and graded afterward. The internal validity of the system was assessed by AUC and calibration plot, and the external validation was performed in the validation dataset. RESULTS: The prevalence rates of NLBP in the development dataset and the validation dataset were 12.97 and 13.27%, respectively. Age, BMI, average exercise times weekly, gender, educational level, the intensity of daily work, place of residence, monthly income, overall evaluation of health condition and physiology health were identified as significant factors. The total risk score ranged from 0 to 38, which was split into three risk grades: low risk (0 to 18), intermediate risk (19 to 22) and high risk (23 to 38). The pre-scoring system had an adequate calibration and a good discriminating ability with bootstrap-corrected AUC equaling 0.861 in the development dataset and 0.821 in the validation dataset. CONCLUSIONS: A pre-scoring system that could help clinicians to assess the risk of NLBP in the general population was validated. Further validation of the system in a new population or prospective cohort study is suggested.

Aspergillus Vertebral Osteomyelitis Complicating Pulmonary Granuloma in an Immunocompetent Adult.

OBJECTIVE: The aim of this report is to describe a case with Aspergillus vertebral osteomyelitis complicating pulmonary granuloma in an immunocompetent adult. CLINICAL PRESENTATION AND INTERVENTION: A 53-year-old male patient was found to have lesions on lumbar vertebra 5 months after thoracoscopic resection of pulmonary granuloma that lacked a definite etiology. Three operations on the lumbar lesions were performed successively; however, an Aspergillus infection was not confirmed until hyphae were clearly detected at the last surgery. The patient was treated with voriconazole and recovered well. CONCLUSION: This case shows that simultaneous occurrence of granulomatous nodules in the lung and vertebral lesions should raise suspicion of aspergillosis, even in immunocompetent patients.

Anterior corpectomy versus posterior laminoplasty for multilevel cervical myelopathy: a systematic review and meta-analysis.

BACKGROUND: Surgical strategy for multilevel cervical myelopathy resulting from cervical spondylotic myelopathy (CSM) or ossification of posterior longitudinal ligament (OPLL) still remains controversial. There are still questions about the relative benefit and safety of direct decompression by anterior corpectomy (CORP) versus indirect decompression by posterior laminoplasty (LAMP). OBJECTIVE: To perform a systematic review and meta-analysis evaluating the results of anterior CORP compared with posterior LAMP for patients with multilevel cervical myelopathy. METHODS: Systematic review and meta-analysis of cohort studies comparing anterior CORP with posterior LAMP for the treatment of multilevel cervical myelopathy due to CSM or OPLL from 1990 to December 2012. An extensive search of literature was performed in Pubmed, Embase, and the Cochrane library. The quality of the studies was assessed according to GRADE. The following outcome measures were extracted: pre- and postoperative Japanese orthopedic association (JOA) score, neurological recovery rate (RR), surgical complications, reoperation rate, operation time and blood loss. Two reviewers independently assessed each study for quality and extracted data. Subgroup analysis was conducted according to the mean number of surgical segments. RESULTS: A total of 12 studies were included in this review, all of which were prospective or retrospective cohort studies with relatively low quality. The results indicated that the mean JOA score system for cervical myelopathy and the neurological RR in the CORP group were superior to those in the LAMP group when the mean surgical segments were <3, but were similar between the two groups in the case of the mean surgical segments equal to 3 or more. There was no statistical difference in the surgical complication rate between the two groups when the mean surgical segments <3, but were significantly higher incidences of surgical complications and complication-related reoperation in the CORP group compared with the LAMP group in the case of the mean surgical segments equal to 3 or more. Besides, the operation time in the CORP group was longer than that in the LAMP group, and the average blood loss was significantly more in the CORP group compared with the LAMP group. CONCLUSION: Based on the results above, anterior CORP and fusion is recommended for the treatment of multilevel cervical myelopathy when the involved surgical segments were <3. Given the higher rates of surgical complications and complication-related reoperation and the higher surgical trauma associated with multilevel CORP, however, it is suggested that posterior LAMP may be the preferred method of treatment for multilevel cervical myelopathy when the involved surgical segments were equal to 3 or more. In addition, taking the limitations of this study into consideration, it was still not appropriate to draw a strong conclusion claiming superiority for CORP or LAMP. A well-designed, prospective, randomized controlled trial is necessary to provide objective data on the clinical results of both procedures.

Identification of Biomarkers, Pathways, Immune Properties of Mitophagy Genes, and Prediction Models for Intervertebral Disc Degeneration.

Background: Intervertebral disc degeneration (IDD) is the leading cause of low back pain (LBP). The mechanism of IDD development and progression is not fully understood. Peripheral biomarkers are increasingly vital non-radioactive methods in early detection and diagnosis for IDD. Nevertheless, less attention has been paid to the role of mitophagy genes in the progress of IDD. This study aimed to identify the mitophagy disease-causing genes in the process of IDD and mitophagy diagnostic biomarkers for IDD. Methods: Mitophagy-related differentially expressed genes (MRDEGs) related to IDD were investigated by analyzing the microarray datasets of IDD cases from GEO, PathCards and Molecular Signatures Databases. We used R software, WGCNA, PPI, mRNA-miRNA, mRNA-TF, GO, KEGG, GSEA, GSVA and Cytoscape to analyze and visualize the data. We further used ssGSEA for immunoinfiltration analysis to obtain different immune cell infiltration. LASSO model was developed to screen for genes that met the diagnostic gene model requirements. Finally, qRT-PCR, Western blotting and HE were used to verify hub genes and their expression from clinical IDD samples. Results: We identified 14 MRDEGs and 12 hub genes. GO, KEGG, GSEA and GSVA analyses demonstrated that hub genes were critical for the development of IDD. LASSO diagnostic model consisted of six hub genes, among which SQSTM1, ATG7 and OPTN were significantly different between the two IDD disease subtypes. At the same time, SQSTM1 also had a high correlation with immune characteristic subtypes. The results of qRT-PCR and Western blotting also indicated that these genes were significantly differentially expressed in nucleus pulposus cells (NPCs) of the IDD group. Conclusion: We explored an association between MRDEGs-associated signature in IDD and validated that hub genes like SQSTM1 might serve as biomarkers for diagnostic and therapeutic targets for IDD. Meanwhile, this study can provide new insights into the functional characteristics and mechanism of mitophagy in the development of IDD.

Comparison of mesenchymal stromal cells from human bone marrow and adipose tissue for the treatment of spinal cord injury.

BACKGROUND AIMS: Bone marrow and subcutaneous adipose tissue are both considered prospective sources of mesenchymal stromal cells (MSCs), which can be used in cell therapy for spinal cord injury (SCI). The present study investigated whether human adipose tissue-derived mesenchymal stromal cells (hADSCs) transplanted into a rat model of SCI would lead to similar or improved neurologic effects compared with human bone marrow-derived mesenchymal stromal cells (hBMSCs). METHODS: hADSCs and hBMSCs were isolated from five adult donors. These MSCs were characterized using flow cytometry, immunocytochemistry, real-time polymerase chain reaction and enzyme-linked immunosorbent assay. Immediately after SCI, 2 × 10(5) hBMSCs or hADSCs were injected into the injured spinal cord. Locomotor function, cell survival and differentiation, spinal cord tissue morphology and brain-derived neurotrophic factor (BDNF) expression were compared between groups. RESULTS: hADSCs and hBMSCs showed similar surface protein expression, and hADSCs showed higher proliferative activity with higher expression of vascular endothelial cell growth factor, hepatocyte growth factor and BDNF than hBMSCs. After transplant, both hADSCs and hBMSCs migrated within the injured spinal cord without differentiating into glial or neuronal elements. Administration of hADSCs was associated with marked changes in the SCI environment, with significant increases in BDNF levels. This was simultaneously associated with increased angiogenesis, preserved axons, decreased numbers of ED1-positive macrophages and reduced lesion cavity formation. These changes were accompanied by improved functional recovery. CONCLUSIONS: The present results suggest that hADSCs would be more appropriate for transplant to treat SCI than hBMSCs.

Effects of Pretreatment of the Dorsal Meningovertebral Ligaments on the Incidence of Intraoperative Cerebrospinal Fluid Leakage: A Comparative Study.

Cerebrospinal fluid leakage can lead to postoperative refractory headaches and meningitis. Dural injury is the main cause of postoperative cerebrospinal fluid leakage. Previously, we performed a comprehensive anatomic study on the dorsal meningovertebral ligaments in the lumbosacral regions and concluded that these ligaments are an anatomic factor leading to dural laceration. However, no clinical study has examined the relationship between dorsal meningovertebral ligaments and the incidence of intraoperative cerebrospinal fluid leakage. The goal of this study was to investigate the effect of prophylactic intraoperative pretreatment of the meningovertebral ligaments on the incidence of cerebrospinal fluid leakage during surgery. [Orthopedics. 2023;46(1):e66-e71.].

Tumor necrosis factor-α inhibition restores matrix formation by human adipose-derived stem cells in the late stage of chondrogenic differentiation.

BACKGROUND: Cartilage tissue engineering is a promising strategy for treating cartilage damage. Matrix formation by adipose-derived stem cells (ADSCs), which are one type of seed cell used for cartilage tissue engineering, decreases in the late stage of induced chondrogenic differentiation in vitro, which seriously limits research on ADSCs and their application. AIM: To improve the chondrogenic differentiation efficiency of ADSCs in vitro, and optimize the existing chondrogenic induction protocol. METHODS: Tumor necrosis factor-alpha (TNF-α) inhibitor was added to chondrogenic culture medium, and then Western blotting, enzyme linked immunosorbent assay, immunofluorescence and toluidine blue staining were used to detect the cartilage matrix secretion and the expression of key proteins of nuclear factor kappa-B (NF-κB) signaling pathway. RESULTS: In this study, we found that the levels of TNF-α and matrix metalloproteinase 3 were increased during the chondrogenic differentiation of ADSCs. TNF-α then bound to its receptor and activated the NF-κB pathway, leading to a decrease in cartilage matrix synthesis and secretion. Blocking TNF-α with its inhibitors etanercept (1 µg/mL) or infliximab (10 µg/mL) significantly restored matrix formation. CONCLUSION: Therefore, this study developed a combination of ADSC therapy and targeted anti-inflammatory drugs to optimize the chondrogenesis of ADSCs, and this approach could be very beneficial for translating ADSC-based approaches to treat cartilage damage.

Comprehensive bioinformatics analysis reveals the hub genes and pathways associated with multiple myeloma.

PURPOSE: While the prognosis of multiple myeloma (MM) has significantly improved over the last decade because of new treatment options, it remains incurable. Aetiological explanations and biological targets based on genomics may provide additional help for rational disease intervention. MATERIALS AND METHODS: Three microarray datasets associated with MM were downloaded from the Gene Expression Omnibus (GEO) database. GSE125364 and GSE39754 were used as the training set, and GSE13591 was used as the verification set. The differentially expressed genes (DEGs) were obtained from the training set, and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed to annotate their functions. The hub genes were derived from the combined results of a protein-protein interaction (PPI) network and weighted gene coexpression network analysis (WGCNA). The receiver operating characteristic (ROC) curves of hub genes were plotted to evaluate their clinical diagnostic value. Biological processes and signaling pathways associated with hub genes were explained by gene set enrichment analysis (GSEA). RESULTS: A total of 1759 DEGs were identified. GO and KEGG pathway analyses suggested that the DEGs were related to the process of protein metabolism. RPN1, SEC61A1, SPCS1, SRPR, SRPRB, SSR1 and TRAM1 were proven to have clinical diagnostic value for MM. The GSEA results suggested that the hub genes were widely involved in the N-glycan biosynthesis pathway. CONCLUSION: The hub genes identified in this study can partially explain the potential molecular mechanisms of MM and serve as candidate biomarkers for disease diagnosis.

Posterior cervical fixation following laminectomy: a stress analysis of three techniques.

The aim of this study was to compare the following three main fixation techniques: pedicle screw (PS) technique, lateral mass screw (LS) technique, and transarticular screw (TS) technique. A detailed, geometrically accurate, nonlinear C3-C7 FE model had been successfully developed and validated. Then three finite element (FE) models were reconstructed by different fixation techniques following C4-C6 level laminectomy. A compressive preload of 74 N combined with a pure moment of 1.8 Nm in flexion, extension, left-right lateral bending, and left-right axial rotation was applied to the models. The results showed that maximum von Mises stress on the fixation devices was much higher in the FE models of TS technique, compared with the models of PS and LS techniques. Furthermore, the screws inserted by TS technique had high stress concentration at the middle part of the screws. Screw inserted by PS and LS techniques had high stress concentration at the actual cap-rod-screw interface. The highest level of maximal stress was obtained with the fixation device of the TS technique. TS technique induces noticeable differences in the stress compared to the posterior cervical fixation technique, regarding the higher stress level on fixation devices.

Structure of Lumbar Intervertebral Foraminal Ligaments Based on 3-Dimensional Reconstruction Through Ultrathin Cryomilling of a Human Cadaver.

BACKGROUND: This study aimed to assess the feasibility of using the 3-dimensional (3-D) reconstruction technique based on ultrathin cryomilling to show the lumbar intervertebral foraminal ligaments in situ. METHODS: Cryomilling was performed on an embalmed human cadaver to acquire successive cross-sectional images. In each of the images, the boundaries of lumbar intervertebral foraminal ligaments and their adjacent structures were outlined, labeled, and reconstructed for 3-D modeling. The morphology, attachments, and spatial orientation of ligaments were described. RESULTS: A total of 9 ligaments in 10 lumbar intervertebral foramina (IVFs) were identified and reconstructed. These ligaments can be divided into 5 types. The IVFs were divided into 2 or 3 main portions by the first 4 types of ligaments (transforaminal ligaments, corporotransverse ligaments, "reticular" ligaments, and "Y-shaped" ligaments). The radiating ligaments (the fifth type of ligaments) attached to the surrounding structures of the IVF and were connected directly to the nerve root sleeves. Although there was no indication of neurovascular compromise in this normal specimen, these ligaments limit the space within the bony IVF such that under certain pathologic conditions (e.g., inflammation), their presence would make neurovascular compression more likely than if they were absent. CONCLUSIONS: The 3-D reconstruction technique based on ultrathin cryomilling can effectively show the lumbar intervertebral foraminal ligaments and their anatomical characteristics in situ, providing a new way to clarify the relationships between these ligaments and their adjacent structures.

Matrix Metalloproteinase 3: A Promoting and Destabilizing Factor in the Pathogenesis of Disease and Cell Differentiation.

Cells must alter their expression profiles and morphological characteristics but also reshape the extracellular matrix (ECM) to fulfill their functions throughout their lifespan. Matrix metalloproteinase 3 (MMP-3) is a member of the matrix metalloproteinase (MMP) family, which can degrade multiple ECM components. MMP-3 can activate multiple pro-MMPs and thus initiates the MMP-mediated degradation reactions. In this review, we summarized the function of MMP-3 and discussed its effects on biological activities. From this point of view, we emphasized the positive and negative roles of MMP-3 in the pathogenesis of disease and cell differentiation, highlighting that MMP-3 is especially closely involved in the occurrence and development of osteoarthritis. Then, we discussed some pathways that were shown to regulate MMP-3. By writing this review, we hope to provide new topics of interest for researchers and attract more researchers to investigate MMP-3.

[Indication of fusion for degenerative lumbar spinal stenosis treated by "windows technique" laminoforaminotomy].

OBJECTIVE: To explore the indications of fusion for degenerative lumbar spinal stenosis treated by "windows technique". METHODS: From December 1999 to December 2005, 145 consecutive patients who were treated by primary decompression with "windows technique" laminoforaminotomy for degenerative lumbar spinal stenosis, a retrospective study, were divided into 3 groups (A and B and C) by preoperative lumbar conditions and surgical methods. In group A, 39 patients with spinal instability or degenerative lumbar spondylolisthesis or scoliosis underwent decompression and fusion; in group B, 31 patients with spinal instability or degenerative lumbar spondylolisthesis or scoliosis underwent decompression alone; In group C, 75 patients without spinal instability or degenerative lumbar spondylolisthesis or scoliosis were treated by decompression without fusion. On hospital medical records to review, they were followed up by telephone and out-patient referral. Statistics the duration of hospitalization, operative time, estimated blood loss; Observed recrudescence and reoperation and complication; and using Oswestry Disability Index and Visual Analog Scale and satisfaction rate for efficacy assessment, application SPSS 13.0 software. RESULTS: All 145 patients had at least a 3-year follow-up (ranging 37 to 108 months). In the group C, the duration of hospitalization less than in the group A or B (P < 0.05); In the group A, the operative time and estimated blood loss greater than in the group B or C (P < 0.05); The group B treated by decompression alone in the presence of instability or spondylolisthesis or scoliosis showed the worst results by the Oswestry Disability Index or Visual Analog Scale or ate of satisfaction (P < 0.05). The same good results can be obtained in the group A and C. There were not different about recrudescence or reoperation or complication in the three groups. CONCLUSIONS: Fusion should be performed on patients with instability or degenerative lumbar spondylolisthesis or scoliosis after primary decompression with "windows technique" laminoforaminotomy. The patient with simple lumbar spinal stenosis undergone primary surgery does not require fusion.

[Degradable performance and bio-mineralization function of PLA-PEG-PLA/PLA tissue engineering scaffold in vitro and in vivo].

The degradable performance and bio-mineralization function of PLA-PEG-PLA/PLA tissue engineering scaffolds in vitro and in vivo were systematically studied. The X-ray diffraction and Fourier transform infrared spectra showed that there was the deposition of bone-like carbonate hydroxyapatite on the surface of scaffolds. We found that the weight of scaffolds did not always decrease with the prolongation of time in vitro. At the same time, we found that after the PLA-PEG-PLA/PLA tissue engineering scaffolds were embedded in skulls of rhesus monkeys, the new bone area reached 75% at the 12th week. Histological observation showed that the new bones were rebuilt and knitted bones were formed at the 12th week. These findings meant that the PLA-PEG-PLA/PLA tissue engineering scaffolds were potential in clinical use.

Therapeutic potential of targeting HSPA5 through dual regulation of two candidate prognostic biomarkers ANXA1 and PSAT1 in osteosarcoma.

Osteosarcoma is the most common primary malignant bone tumor that mostly affects young people's health. The prognosis of patients with unresectable or recurrent osteosarcoma still remains dismal. Based on gene integration analysis from GEO and TARGET databases by R language, the differentially expressed genes of osteosarcoma patients were identified. Biological molecular function analysis indicated that these genes were importantly enriched in the process of cell adhesion molecule binding. Gene significance highly-related to clinical traits of osteosarcoma was found by weighted gene co-expression network analysis. Additionally, receiver operating characteristic curve analysis was conducted to find prognostic markers in LASSO Cox regression model. Two candidate biomarkers, ANXA1 and PSAT1, for the prognosis of osteosarcoma were detected separately on the basis of WGCNA and LASSO model. Of note, their expression profiles were interrelated with an important therapeutic target HSPA5. In vitro pharmaceutical experiments were performed to explore the biological role and prognostic benefit of candidates. Suppression of HSPA5 effectively upregulated ANXA1 and inhibited PSAT1, resulting in osteosarcoma cell proliferation arrest and apoptosis. These findings suggest that HSPA5 serves as a core molecule for osteosarcoma therapy due to its bidirectional regulation of candidate prognostic biomarkers ANXA1 and PSAT1.

[Vascular endothelial growth factor/polylactide-polyethyleneglycol-polylactic acid copolymer/basic fibroblast growth factor mixed microcapsules in promoting angiogenic differentiation of rat bone marrow mesenchymal stem cells in vitro].

Objective: To observe the effect of vascular endothelial growth factor/polylactide-polyethyleneglycol-polylactic acid copolymer/basic fibroblast growth factor (VEGF/PELA/bFGF) mixed microcapsules in promoting the angiogenic differentiation of rat bone marrow mesenchymal stem cells (BMSCs) in vitro. Methods: The BMSCs were isolated by the method of whole bone marrow adherent, and sub-cultured. The passage 3 BMSCs were identified by Wright-Giemsa staining and flow cytometry, and used for subsequent experiments. VEGF/PELA/bFGF (group A), PELA/bFGF (group B), VEGF/PELA (group C), and PELA (group D) microcapsules were prepared. The biodegradable ability and cytotoxicity of PELA microcapsule were determined，and the slow-released ability of VEGF/PELA/bFGF mixed microcapsules was measured. The passage 3 BMSCs were co-cultured with the extracts of groups A, B, C, and D, separately. At 1, 3, 7, 14, and 20 days after being cultured, the morphological changes of induced BMSCs were recorded. At 21 days, the induced BMSCs were tested for DiI-labeled acetylated low density lipoprotein (Dil-ac-LDL) and FITC-labeled ulex europaeus agglutinin I (FITC-UEA-I) uptake ability. The tube-forming ability of the induced cells on Matrigel was also verified. The differences of the vascularize indexes in nodes, master junctions, master segments, and tot.master segments length in 4 groups were summarized and analyzed. Results: The isolated and cultured cells were identified as BMSCs. The degradation time of PELA was more than 20 days. There was no significant effect on cell viability under co-culture conditions. At 20 days, the cumulative release of VEGF in the mixed microcapsules exceeded 95%, and the quantity of bFGF exceeded 80%. The morphology of cells in groups A, B, and C were changed. The cells in groups A and B showed the typical change of cobble-stone morphology. The numbers of double fluorescent labeled cells observed by fluorescence microscope were the most in group A, and decreases from group B and group C, with the lowest in group D. The cells in groups A and B formed a grid-like structure on Matrigel. Quantitative analysis showed that the differences in the number of nodes, master junctions, master segments, and tot.master segments length between groups A, B and groups C, D were significant ( P<0.05). The number of nodes and the tot.master segments length of group A were more than those of group B ( P<0.05). There was no significant differences in the number of master junctions and master segments between group A and group B ( P>0.05). Conclusion: VEGF/PELA/bFGF mixed microcapsules have significantly ability to promote the angiogenic differentiation of rat BMSCs in vitro.

MiR-27a promotes the autophagy and apoptosis of IL-1ß treated-articular chondrocytes in osteoarthritis through PI3K/AKT/mTOR signaling.

Osteoarthritis (OA) is a common degenerative joint disorder, which involves articular cartilage degeneration as well as joint inflammatory reactions. The recent studies have identified microRNA (miRNA) as one of the epigenetic mechanisms for the regulation of gene expression. Here we aim to reveal the role of miRNA in the regulation of gene expression in articular chondrocytes and its significance in the OA pathogenesis. In the present study, miRNA profiling was performed using OA cartilage and normal healthy cartilage tissues. As compared to their levels in normal cells and tissues, miR-27a expression was found to be upregulated in OA cartilage and IL-1ß-treated articular chondrocytes. TUNEL staining, as well as flow cytometry with Annexin V-FITC/PI double labeling indicated that miR-27a inhibition reduced the apoptosis of IL-1ß-treated articular chondrocytes. Bioinformatics prediction and the dual-luciferase reporter assay indicated that miR-27a targeted the 3'-UTR of the PI3K gene to silence it. The PI3K mRNA level in OA cartilage and IL-1ß-treated articular chondrocytes was also downregulated, comparing with normal cells and tissues. Transfection of chondrocytes transfected with the miR-27a inhibitor upregulated the PI3K expression. This study demonstrated miR-27a is a regulator of the PI3K-Akt-mTOR axis in human chondrocytes and could participate in OA pathogenesis.