MFNG is an independent prognostic marker for osteosarcoma.

BACKGROUND: Osteosarcoma (OS) has been the most common malignancy of the bone in children and adolescents, and the unsatisfactory prognosis of OS sufferers has long been a hard nut. Here, we delved into the markers with a prognostic value for predicting the prognosis of OS patients. METHODS: The messenger RNA (mRNA) sequencing data and clinical data of OS were retrieved from a Gene Expression Omnibus (GEO) dataset (GSE39058). Next, prognosis-related genes (PRGs) were filtered with the aid of Kaplan-Meier (K-M) curves and Cox regression analysis (CRA). Later, Gene Ontology (GO) biological process analysis was used in verifying the function of different genes. CCK-8 and cell apoptosis assay were performed to evaluate the function of MFNG in U2OS cells. RESULTS: Among the obtained genes, Manic Fringe (MFNG) had the closest relevance to prognosis and clinical traits, thus becoming the research object herein. In light of the expression level of MFNG, patients fell into high- and low-MFNG groups. Patients with elevated MFNG expression had a worse prognosis, according to the survival analysis. It was unveiled by the univariate and multivariate analyses that MFNG expression was an independent adverse prognostic factor for disease-free survival in OS patients (p = 0.006). Meanwhile, MFNG expression was linked to gender and tumor recurrence, and it was higher in patients with OS recurrence. Moreover, overexpression of MFNG promoted the cell proliferation and inhibited the cell apoptosis of U2OS cells. CONCLUSIONS: The expression level of MFNG negatively correlated with OS progression, and as an independent adverse prognostic factor for disease-free survival in OS patients. Moreover, MFNG regulated the cell proliferation and apoptosis of OS cells.

A/(H1N1) pdm09 NS1 promotes viral replication by enhancing autophagy through hijacking the IAV negative regulatory factor LRPPRC.

The quadrilateral reassortant IAV A/(H1N1) pdm09 is the pathogen responsible for the first influenza pandemic of the 21st century. The virus spread rapidly among hosts causing high mortality within human population. Efficient accumulation of virions is known to be important for the rapid transmission of virus. However, the mechanism by which A/(H1N1) pdm09 promotes its rapid replication has not been fully studied. Here, we found the NS1 of A/(H1N1) pdm09 mediated complete macroautophagy/autophagy, and then facilitated self-replication, which may be associated with the more rapid spread of this virus compared with H1N1WSN and H3N8JL89. We found that the promotion of self-replication could be mainly attributed to NS1pdm09 strongly antagonizing the inhibitory effect of LRPPRC on autophagy. The interaction between NS1pdm09 and LRPPRC competitively blocked the interaction of LRPPRC with BECN1/Beclin1, resulting in increased recruitment of BECN1 for PIK3C3 (phosphatidylinositol 3-kinase catalytic subunit type 3) and induction of the initiation of autophagy. In conclusion, we uncover the unique molecular mechanism by which A/(H1N1) pdm09 utilizes autophagy to promote self-replication, and we provide theoretical basics for the analysis of the etiological characteristics of the A/(H1N1) pdm09 pandemic and the development of anti-influenza drugs and vaccines.Abbreviations: 293T: human embryonic kidney 293 cells; 293T_LRPPRC: stable LRPPRC expression 293T cells; 3-MA: 3-methyladenine; A549 cells: human non-small cell lung cancer cells; AA: amino acid; ACTB: actin beta; BECN1: beclin 1; BECN1 KO: BECN1 knockout 293T cells; Cal: calyculin A; Co-IP: co-immunoprecipitation; CQ: chloroquine; DC: dendritic cell; Eug: eugenol; GFP: green fluorescent protein; HA: hemagglutinin; HIV: human immunodeficiency virus; IAVs: Influenza A viruses; IFN: interferon; JL89: A/equine/Jilin/1/1989 (H3N8); LAMP2: lysosomal associated membrane protein 2; LRPPRC: leucine rich pentatriicopeptide repeat containing; LRPPRC KO: LRPPRC knockout 293T cells; M2: matrix 2; MAP1LC3B/LC3B: microtubule associated protein 1 light chain 3 beta; MDCK: Madin-Darby canine kidney cells; MOI: multiplicity of infection; MS: mass spectrometry; NP: nucleoprotein; NS1: non-structural protein 1; NS1JL89: non-structural protein 1 of A/equine/Jilin/1/1989 (H3N8); NS1pdm09: non-structural protein 1 of A/(H1N1) pdm09; NS1SC09: non-structural protein 1 of A/Sichuan/2009 (H1N1); NS1WSN: non-structural protein 1 of A/WSN/1933 (H1N1); PB1: polymerase basic protein 1; PB1-F2: alternate reading frame discovered in PB1 gene segment; PIK3C3: phosphatidylinositol 3-kinase catalytic subunit type 3; PR8: A/PR/8/34 (H1N1); Rapa: rapamycin; RFP: red fluorescent protein; SC09: A/Sichuan/2009 (H1N1); SQSTM1/p62: sequestosome 1; STK4/MST1: serine/threonine kinase 4; TEM: transmission electron microscopy; TOMM20: translocase of outer mitochondrial membrane 20; WHO: World Health Organization; WSN: A/WSN/1933 (H1N1); WSN-NS1JL89: WSN recombinant strain in which NS1 was replaced with that of JL89; WSN-NS1SC09: WSN recombinant strain in which NS1 was replaced with that of SC09.

Methylation recognition protein YTH N6-methyladenosine RNA binding protein 1 (YTHDF1) regulates the proliferation, migration and invasion of osteosarcoma by regulating m6A level of CCR4-NOT transcription complex subunit 7 (CNOT7).

N6-methyladenosine (m6A) is one of the most significant modifications in human mRNAs. Emerging evidence indicates that m6A participates in the initiation and development of malignant tumors. Nevertheless, the biological roles and mechanism of m6A in osteosarcoma (OS) remain unclear. The purpose of this study was to investigate the role and mechanism of the methylation recognition protein-YTH N6-methyladenosine RNA binding protein 1 (YTHDF1) in OS. The YTHDF1 expression in OS was detected by qRT-PCR and Western blot assay. M6A quantification was utilized to measure the methylation level of OS. Cell counting kit-8 (CCK8), 5-Ethynyl-2'-deoxyuridine (EdU) assay and transwell experiments were conducted to confirm the biological effects of YTHDF1 on OS cells. The bioinformatics websites and in vitro assays were conducted to analyze the downstream targets of YTHDF1 was upregulated in OS tissues at mRNA and protein level. The results showed that the expression level of YTHDF1 might be closely associated with the poor prognosis for OS patients. Inhibition of YTHDF1 could suppress the proliferation, migration and invasion of the OS cells. Moreover, we found that CCR4-NOT transcription complex subunit 7 (CNOT7) might be the potential target of YTHDF1, which was upregulated in OS tissues. YTHDF1 could recognize the m6A sites of CONT7 and promote its expression in an m6A manner. Moreover, methyltransferase-like 3 (METTL3) could promote the m6A level of CONT7. YTHDF1 was upregulated in OS and could promote cell proliferation, migration and invasion. The METTL3-CONT7-YTHDF1 regulatory axis might be the potential target for the prognosis and therapy of OS.

ERO1α inhibits cell apoptosis and regulates steroidogenesis in mouse granulosa cells.

ER oxidoreduclin 1α (ERO1α), an oxidase that exists in the ER, participates in protein folding and secretion and inhibiting apoptosis, and regulates tumor progression, which is a novel factor of poor cancer prognosis. However, the other physiological functions of ERO1α remain undiscovered. Although our preliminary results of this study indicated that ERO1α revealed the robust expression in ovary, especially in granulosa cells, the role of ERO1α in follicular development is not well known. Therefore, the aims of the present study were to explore the role of ERO1α and the possible mechanisms in regulating cell apoptosis and steroidogenesis in ovarian granulosa cells. ERO1α was mainly localized in granulosa cells and oocytes in the adult ovary by immunohistochemistry. Western blot analysis showed that the expression of ERO1α was highest at oestrous stage during the estrous cycle. The effect of ERO1α on cell apoptosis and steroidogenesis was detected by transduction of ERO1α overexpression and knockdown lentiviruses into primary cultured granulosa cells. Flow cytometry analysis showed that ERO1α decreased granulosa cells apoptosis. Western bolt and RT-qPCR analysis found that ERO1α increased the ratio of BCL-2/BAX, and decreased BAD and Caspase-3 expression. ELISA analysis showed that ERO1α enhanced estrogen (E2) secretion. Western bolt and RT-qPCR analysis found that ERO1α increased StAR, CYP11A1, 3ß-HSD, CYP17A1, and CYP19A1 expression, and decreased CYP1B1 expression. Furthermore, Western bolt analysis found that ERO1αincreased PDI and PRDX 4 expression, and activated the PI3K/AKT/mTOR signaling pathway through increasing the phosphorylation of AKT and P70 S6 kinase. In summary, these results suggested that ERO1α might play an anti-apoptotic role and regulate steroidogenesis in granulosa cells, at least partly, via activation of the PI3K/AKT/mTOR signaling pathway.

MiR-20a inhibits the progression of human arthritis fibroblast-like synoviocytes and inflammatory factor expression by targeting ADAM10.

MiR-20a has been reported as a key regulator to pro-inflammatory factor release in fibroblast-like synoviocytes (FLS), which caused rheumatoid arthritis (RA). However, the molecular mechanism of miR-20a in RA remains to be further elucidated. This study aimed to investigate the roles of miR-20a in RA pathology. RA (n = 24) and osteoarthritis (OA, n = 20) and normal healthy tissues (n = 16) were collected from operation. TargetScan and dual-luciferase reporter were performed to predict and confirm the potential binding sites of miR-20a on ADAM metallopeptidase domain 10 (ADAM10). Pearson's analysis was adopted to evaluate the correlation between miR-20a and ADAM10 expression. It was found that MiR-20a was downregulated in RA tissues, and overexpressed miR-20a inhibited cell viability, migration and invasion, and the expression of inflammatory factors in RA-FLS MH7A cells. ADAM10 was identified as the target gene of miR-20a, and upregulation of ADAM10 reversed the inhibitory effects of miR-20a. In conclusion, miR-20a inhibits the progression of RA-FLS as well as the inflammatory factor expression by targeting ADAM10.

Puerarin alters the function of monocytes/macrophages and exhibits chondroprotection in mice.

Recent studies have suggested that puerarin may impede osteoclastogenesis and facilitate bone regeneration, in addition to attenuating tissue inflammation. The present study investigated the therapeutic effects of puerarin on inflammatory responses and monocyte recruitment in in vitro and in vivo osteoarthritis (OA) models. Puerarin treatment increased the proliferation of OA chondrocytes, as determined by Cell Counting Kit­8 assay. In addition, the present results suggested that puerarin suppressed the interleukin­1ß­induced production of inflammatory cytokines in OA chondrocytes and monocytes/macrophages, as assessed by ELISA. In a mouse model of mono­iodoacetate­induced OA, the present histological analyses suggested that administration with puerarin attenuated the inflammatory profile of OA joints and reduced cartilage destruction. Using flow cytometry, a decreased number of myeloid­derived C­C chemokine receptor 2+/lymphocyte Ag 6C+ monocytes was identified in the blood of OA mice treated with puerarin compared with control OA mice. Furthermore, quantitative real­time polymerase chain reaction analysis suggested that puerarin treatment decreased C­C chemokine ligand 2 expression in arthritic tissues. Collectively, the results suggested that puerarin treatment limited the recruitment of inflammatory monocytes. In summary, the present study provided pre­clinical evidence that puerarin may serve as a potential target in the treatment of OA.

SIRT6 inhibits proliferation and invasion in osteosarcoma cells by targeting N-cadherin.

SIRT6, is a member of the NAD-dependent sirtuin family of enzymes, and has been reported as a novel tumor suppressor gene or oncogene, dependent on the type of cancer. However, the role of SIRT6 in osteosarcoma has not been investigated. The present study demonstrated that the expression of SIRT6 was downregulated in osteosarcoma tissues and osteosarcoma cell lines when compared with adjacent tissues or osteoblastic cell lines. Kaplan-Meier analysis was performed to evaluate the prognostic significance of SIRT6. The overall survival of patients with higher expression of SIRT6 was significantly longer than patients with lower expression. Subsequently, MTT and invasion assays were performed to detect the biological functions of SIRT6 in osteosarcoma cells in vitro. The results revealed that overexpression of SIRT6 inhibited SAOS-2 and MG-63 cell proliferation and invasion. Knockdown of SIRT6 enhanced cell ability for the proliferation and invasion. A qChIP assay, luciferase reporter assay, reverse transcription-quantitative polymerase chain reaction and western blotting confirmed that CDH2 (N-cadherin) was a target of SIRT6. SIRT6 overexpression suppressed N-cadherin on the mRNA and protein levels. In addition, it was confirmed that the promotional effect of Si-SIRT6 on OS cell growth and invasion was suppressed by downregulating N-cadherin. The present study suggested that SIRT6 may serve as a tumor suppressor during the development of osteosarcoma. In addition, N-cadherin may be a promising therapeutic target for osteosarcoma.

Efficacy of pre-emptive use of cyclooxyenase-2 inhibitors for total knee arthroplasty: a mini-review.

Total knee arthroplasty (TKA) is regarded as the most effective surgery for patients with later-stage arthritis of the knee, but the postoperative pain management for functional improvement of the knew is still a challenging task. This review discusses the mechanism by which the selective cyclooxyenase-2 inhibitors, which reduce the peripheral and central sensitization, decrease pain after TKA. This review also covers the protocols, safety, efficacy, and progress of cyclooxyenase-2 inhibitors in pre-emptive analgesia.

PU.1 attenuates TNF­α­induced proliferation and cytokine release of rheumatoid arthritis fibroblast­like synoviocytes by regulating miR­155 activity.

The present study aimed to determine the role of transcription factor PU.1 (PU.1) in tumor necrosis factor­α (TNF­α)­induced proliferation and cytokine release of rheumatoid arthritis fibroblast­like synoviocytes (RA­FLS). It was determined that TNF­α induced proliferation of RA­FLS, whereas transfection with PU.1 3'untranslated region (UTR) inhibited this proliferation. Additionally, PU.1 3'UTR attenuated TNF­α­induced production of interleukin (IL)­6 and IL­1ß, and downregulated the expression level of micro RNA (miR)­155 in a dose­dependent manner. Furthermore, transfection with PU.1 3'UTR significantly attenuated TNF­α­induced decrease in forkhead box protein O3 (FOXO3) expression level in RA­FLS and these effects were consistent with the effects of miR­155 inhibition. PU.1 and FOXO3 formed a competing endogenous RNA (ceRNA) network that regulated miR­155 activity. In this competing endogenous RNA network, PU.1 3'UTR modulated FOXO3 expression in a miRNA­ and 3'UTR­dependent manner. Downregulation of FOXO3 expression reversed the PU.1 3'UTR­mediated protective effects. Therefore, the results of the present study indicate that PU.1 3'UTR attenuates TNF­α­induced proliferation and cytokine release of RA­FLS by acting as a ceRNA for FOXO3 to regulate miR­155 activity.

miR-149 promotes human osteocarcinoma progression via targeting bone morphogenetic protein 9 (BMP9).

OBJECTIVES: To investigate the roles of miR-149 in the progression of human osteosarcoma (OS). RESULTS: miR-149 level was upregulated in tissues from OS patients more than in normal subjects. Cell proliferation and apoptosis assays revealed that miR-149 increased cell proliferation and inhibited cell apoptosis in OS cell line (MG63). An increase of Bcl-2 gene expression and a decrease of cleaved-caspase-3, and cleaved-PARP expression were observed in MG63 cells with transfection of miR-149. Additionally, bone morphogenetic protein 9 (BMP9) was identified as a target of miR-149 in MG63 cells, and BMP9 expression was negatively correlated with miR149 level in OS clinical samples. Co-overexpression of BMP9 with miR-149 in MG63 cells prohibited miR-149-mediated promotive effects on OS progression. Importantly, overexpression of miR-149 conferred chemoresistance in MG63 cells. CONCLUSIONS: miR-149 promotes OS progression via targeting BMP9.

Results of a hydroxyapatite-coated femoral stem (Corail) in Chinese: a minimum 10-year follow-up.

BACKGROUND: Due to the adverse effects of cemented hip arthroplasty, uncemented stems with hydroxyapatite (HA) coating reduces these risks and enhanced integration. The concept of an extensive HA coating for the fixation of a tapered femoral stem (Corail®) was introduced, which can achieve durable biological fixation and preserve normal periprosthetic bone activity. Here we describe the clinical and radiological outcome in patients with the Corail® stem. METHODS: 92 total hip replacements in 81 patients using the Corail® stem were followed-up. 47 patients were women, and the mean age at surgery was 62.9 ± 8.7 (34-71) years. The indications included: osteoarthritis of the hip (71.1%), avascular necrosis (13.6%), femur neck fractures in elderly (9.7%) and post-traumatic osteoarthritis (6.8%). FINDINGS: Eight patients died during follow-up. The revision was only found in two patients due to line wear and resulted in an 10-year Kaplan-Meier estimated overall survival rate of 97.83%. The clinical results were good, with a mean Harris hip score of 92.3 ± 5.6 (72-100). The mean total Merle d'Aubigné and Postel score was 6.8 ± 0.5 pre-operatively and 16.1 ± 1.4 at latest follow-up. All unrevised implants were radiographically stable, with a mean liner wear of 0.07 mm/year. CONCLUSION: This long-term analysis confirmed the durability of the functional and radiographic results. Our findings suggest the long-term results of Corail® HA-coated stem are more satisfactory which is preferable to any other system.

Mid-term results of metal-on-metal hip resurfacing for treatment of osteoarthritis secondary to developmental dysplasia of the hip: a minimum of 8-years of follow-up.

BACKGROUND: Metal-on-metal resurfacing arthroplasty is an attractive alternative to conventional total hip arthroplasty in patients with osteoarthritis secondary to developmental dysplasia of the hip (DDH). The purpose of this study was to assess the mid-term clinical outcome and mid-term survivorship of Metal-on-metal resurfacing arthroplasty in patients suffering from osteoarthritis secondary to DDH. MATERIAL/METHODS: Between May 2003 and Dec. 2005, 15 operations using ASR™ and 19 using Corin were performed in 29 patients to treat advanced osteoarthritis secondary to DDHs. There were 6 males (20.7%) and 23 females (79.3%), with an average age of 47.2 years (range, 36-64 years). Clinical and radiographic results were observed. All patients were followed up at the 1st, 2nd, 3rd, 6th, and 12th months after surgery and annually thereafter. RESULTS: The overall survival was 88.2% at a minimum follow-up of 8 years, but the survival was 91.2% after excluding the infections as the cause of component loosening and failure. The mean Harris hip score improved from 48.27±3.13 (range, 14-71) to 89.63±3.42 (range, 65-100) at latest follow-up. The flexion was from 75.14±8.05° to 107.21±9.34. Only 4 failed because of deep infection, femoral neck fracture, and aseptic loosening. CONCLUSIONS: Metal-on-metal resurfacing arthroplasty showed perfect results at a minimum of 8-years of follow-up in our study, and may be a reasonable option for osteoarthritis secondary to developmental dysplasia of the hip (DDH).

Granulocytic sarcoma, an undiagnosed leukemia, initially manifested as paralysis.

Granulocytic sarcoma (GS) may be a presenting sign of myelogenous leukemia. Occasionally, an extramedullary neoplasm composed from myelocytic precursor cells occurs in patients without evidence of leukemia. Rarely, undiagnosed leukemia occurs initially manifesting with paralysis to spinal cord GS. We present a case report of 20-year-old girl with an undiagnosed leukemia, initially manifesting as paralysis. En bloc spondylectomy with chemotherapy postoperatively constituted the treatment of choice for this tumor. After two courses of chemotherapy, the patient made a good postoperative recovery with notable bilateral lower extremity improvement.

[Total hip surface replacement arthroplasty: report of 31 cases].

OBJECTIVE: To introduce the technique of total hip surface replacement, evaluate the early results and review the factors which affect the results. METHODS: From October 2000 to January 2005, 31 patients (37 hips) with osteonecrosis, osteoarthritis, hip dysplasia, ankylosing spondylitis were treated with the total hip surface replacement. Among them, 15 were male, and 16 were female, with an average age of 42 years (range from 23 - 65 years). All the 31 patients had the indications for hip surface replacement. Standard operation technique which was brought forth by Amstutz and Nelson was employed, and all patients were followed up after operation. RESULTS: Patients were followed up for an average period of 42 months (3 - 51 months). There were no femoral neck fracture, no dislocation, no infection in all patients. Radiolucent line existed around acetabular prosthesis in 1 hip and another hip had been revised because the prosthesis of femoral head was in incorrect situation. The average Harris hip score improved significantly from 30 to 90, and the score was 93 in the latest follow-up. Based on Harris system, 35 hips were excellent, 1 hip good, and 1 hip fail. CONCLUSIONS: The total hip surface replacement is an effective solution for the problem of the patients with osteonecrosis of the femoral head, osteoarthritis, hip dysplasia and ankylosing spondylitis. The short-term results are satisfied.

[Resurfacing arthroplasty for treatment of avascular necrosis of femoral head in young and middle-aged patients].

OBJECTIVE: To investigate the early clinical effect of resurfacing arthroplasty on the treatment of avascular necrosis of the femoral head in the young and middle-aged patients. METHODS: Eleven patients with avascular necrosis of the femoral head in Ficat Stages III -IV (14 hips) were treated by femoral head resurfacing operations. Of 11 cases, there were 7 males and 4 females. With an age range of 35 to 49 years. While 13 patients with avascular necrosis of the femoral head in Ficat Stages III - IV (16 hips) were treated by total hip resurfacing arthroplasty of 13 cases there were 8 males and 5 females. With an age range of 23 to 48 years. The prostheses were improved in light of the anatomic features of the Chinese. RESULTS: These patients treated by femoral head resurfacing operations were followed up for 1 to 5 years. The average Harris hip score was increased from 39 points preoperatively to 91 points postoperatively. These patients treated by total hip resurfacing operations were followed up for 6 to 40 months. The average Harris hip score was increased from 30 points preoperatively to 93 points postoperatively. CONCLUSION: Hip resurfacing operations may be the most effective treatment for avascular necrosis of the femoral head in the young and middle aged patients.