The SUMO-interacting motif in NS2 promotes adaptation of avian influenza virus to mammals.

Species differences in the host factor ANP32A/B result in the restriction of avian influenza virus polymerase (vPol) in mammalian cells. Efficient replication of avian influenza viruses in mammalian cells often requires adaptive mutations, such as PB2-E627K, to enable the virus to use mammalian ANP32A/B. However, the molecular basis for the productive replication of avian influenza viruses without prior adaptation in mammals remains poorly understood. We show that avian influenza virus NS2 protein help to overcome mammalian ANP32A/B-mediated restriction to avian vPol activity by promoting avian vRNP assembly and enhancing mammalian ANP32A/B-vRNP interactions. A conserved SUMO-interacting motif (SIM) in NS2 is required for its avian polymerase-enhancing properties. We also demonstrate that disrupting SIM integrity in NS2 impairs avian influenza virus replication and pathogenicity in mammalian hosts, but not in avian hosts. Our results identify NS2 as a cofactor in the adaptation process of avian influenza virus to mammals.

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.

Taraxasterol inhibits inflammation in osteoarthritis rat model by regulating miRNAs and NF-κB signaling pathway.

Osteoarthritis (OA) has a high incidence rate in the elderly population and is a cause of chronic degenerative joint disease. Current therapeutic approaches to OA are effective but come with some side effects. Therefore, it is urgent to find new safe and effective OA treatments. This study aimed to clarify the function of taraxasterol (TAX) isolated from Taraxacum officinale in the papain-induced rat OA model. We observed that TAX alleviated the typical OA-caused phenomena in the joint. The expression of serum inflammatory mediators such as TNF-α, IL-6, and IL-1ß was also repressed by TAX. In addition, NF-κB signaling pathway was repressed by TAX. Furthermore, two microRNAs: miR-140 and miR-146a were elevated after TAX treatment in OA rat model. Interestingly, several common targets of miR-140 and miR-146a, including HSPA4L, ST5, and ERBB4, were confirmed to be regulated by TAX. Inflammatory response related genes including S100A8, CCL3, A2M, LBP, and CCR1 were repressed by TAX in OA rat model. In summary, TAX inhibits inflammation in osteoarthritis rat model. Inflammatory mediators, NF-κB pathway and miR-140/miR-146a targets mediate the function of TAX.

[Purification of influenza C virus polymerase based on huANP32A protein and efficient preparation of monoclonal antibody targeting PB2 protein].

Influenza C virus is an important respiratory pathogen not only infecting people, but also pigs, dogs, and other animals. Polymerase is central to the replication of influenza C virus and is an important target for studying the mechanism of viral replication. However, there is no commercial monoclonal antibody (MAb) targeting influenza C virus polymerase, which hampers the development of relevant research to some extent. In order to prepare MAb targeting the polymerase basic protein 2 (PB2) of influenza C virus, influenza C virus RNA-dependent RNA polymerase (RdRp, consists of PB1, PB2 and P3) was co-immunoprecipitated with Flag-tagged human acidic nuclear phosphoprotein 32A (huANP32A-Flag) from 293T cells based on the interaction between huANP32A and influenza virus RdRp. The purified RdRp was used as antigen to immunize BALB/c mice. Six positive hybridoma cell lines (7B11-5, 8A4-5, 13D9-6, 8D4-1, 8D4-3, 9F9-4) that stably secrete and recognize PB2 MAb were screened by indirect ELISA and Western blotting. The subtypes of MAb 7B11-5, 8A4-5, 8D4-1 and 8D4-3 antibody were identified as IgG1, the subtypes of MAb 13D9-6 and 9F9-4 were IgG2a and IgG3, respectively. All the light chains of the MAbs were κ chain. A hybridoma cell line 8D4-1 with high titer was further selected to prepare ascites. The titer of mouse ascites antibody was determined to be 1:64 000. Western blotting results showed that the MAb 8D4-1 had a specific immune response with ICV PB2; laser confocal assay showed that the prepared MAb 8D4-1 accurately detected the subcellular localization of PB2 subunits. Moreover, ICV RdRp was highly enriched by ANP32A. The high specific of the prepared PB2 MAb 8D4-1 may facilitate the polymerase detection, structural analysis and mechanism study of influenza C virus.

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.

Comparison of Efficacy of Teriparatide (Parathyroid Hormone 1-34) Alone and in Combination with Zoledronic Acid for Osteoporosis in Postmenopausal Women.

The purpose of this study was to compare the efficacy of teriparatide (parathyroid hormone 1-34) alone and in combination with zoledronic acid (ZA) for the treatment of osteoporosis in postmenopausal women. Ninety-six patients were randomly equally divided into Groups A (n=48) and B (n=48). Group A was given parathyroid hormone 1-34 alone. Group B was treated with parathyroid hormone 1-34 plus ZA. Visual analogue scale (VAS) score, bone mineral density(BMD), serum osteopontin (OPN), and C-terminal cross-linking telopeptide of type I collagen (S-CTX) etc. were compared. After 6 months of treatment, VAS score, serum OPN and S-CTX levels in Group B were significantly lower than those in Group A (p=0.001, p<0.001 and p<0.001, respectively); and BMD values of lumbar vertebrae L2-4, femoral neck and total hip bone in Group B were higher than those of Group A (p=0.002, p=0.028 and p<0.001, respectively). In conclusion, parathyroid hormone 1-34 plus ZA is more effective than parathyroid hormone 1-34 alone in treating post postmenopausal osteoporosis. Key Words: Teriparatide (parathyroid hormone 1-34, Zoledronic acid (ZA), Postmenopausal, Osteoporosis, Bone mineral density (BMD).

Denosumab might prevent periprosthetic bone loss after total hip and knee arthroplasties: a review.

Total hip arthroplasty and total knee arthroplasty are extensively used for the treatment of the end-stage degenerative joint diseases. Currently, periprosthetic bone loss is still the major cause of aseptic loosening, resulting in implant failures. Previous literature introduced some widely accepted protocols for the prevention and treatment of periprosthetic bone loss, but no guideline has been proposed. Denosumab, a human monoclonal immunoglobulin G2 (IgG2) antibody, can inhibit bone resorption by binding to the receptor activator of nuclear factor kappa-B ligand (RANKL). This article reviews the present findings and evidence concerning the effect of denosumab on the periprosthetic bone loss after total hip arthroplasty and total knee arthroplasty. Overall, the current evidence suggests that denosumab is a promising agent for the treatment of periprosthetic bone loss.

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.

Effect of Intra-articular Ketorolac Versus Corticosteroid Injection for Knee Osteoarthritis: A Retrospective Comparative Study.

BACKGROUND: Intra-articular corticosteroid injections have been widely used and are considered a mainstay in the nonoperative treatment of symptomatic knee osteoarthritis (OA). However, their increased use can have negative implications, including chondral toxicity and a high risk of infections. As a result, nonsteroidal anti-inflammatory drugs have been considered as an alternative. PURPOSE: To determine the pain relief and safety of ketorolac versus a corticosteroid to supplement an intra-articular sodium hyaluronate injection for the treatment of symptomatic knee OA. STUDY DESIGN: Cohort study; Level of evidence, 3. METHODS: A total of 84 patients with unilateral symptomatic knee OA receiving 5 weekly injections were enrolled in this retrospective study. Group A (n = 42) received 3 weekly intra-articular corticosteroid injections (0.5% lidocaine, 25 mg of triamcinolone acetonide, and 25 mg of sodium hyaluronate, followed by 2 weekly injections of 0.5% lidocaine and 25 mg of sodium hyaluronate), while group B (n = 42) received 5 weekly ketorolac injections (0.5% lidocaine, 10 mg of ketorolac, and 25 mg of sodium hyaluronate). The following parameters were used to evaluate pain relief and safety: visual analog scale (VAS) for pain, Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), and side effects before the injection and at 1, 2, and 5 weeks after treatment commencement as well as 3 months after the last injection. RESULTS: Patients from both groups had a significant improvement in VAS and WOMAC scores from the first injection to final follow-up at 3 months. In the first week, the VAS score was lower in group A (P = .041), but no significant between-group differences were found for either the VAS or the WOMAC score at the other time points. Of the 42 patients in group A, 34 (81.0%) and 25 (59.5%) achieved successful outcomes at 5 weeks after treatment commencement and 3 months after the last injection, respectively. In group B, 32 (76.2%) and 24 (57.1%) patients achieved successful outcomes at 5 weeks after treatment commencement and 3 months after the last injection, respectively. At final follow-up, no significant difference was found in the successful treatment rate between the groups (P = .825). CONCLUSION: The current study demonstrated that intra-articular ketorolac and corticosteroid injections produce the same pain relief and functional improvement.

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.

Treatment of persistent hiccups after arthroplasty: effects of acupuncture at PC6, CV12 and ST36.

OBJECTIVE: There are multiple treatment options for hiccups, including non-pharmacological therapies, but little evidence of superiority of one treatment over another. The aim of this study was to investigate the effects of acupuncture on persistent hiccups after arthroplasty. METHODS: From April 2010 to December 2015, 15 patients with primary unilateral total hip/knee arthroplasty were diagnosed with persistent hiccups and given acupuncture at PC6, CV12 and ST36. Each acupuncture session lasted 30 min. The total number of treatment sessions was determined by the persistence of symptoms, but acupuncture was administered no more than three times over the course of a week. The hiccups assessment instrument (HAI) was used to assess the severity of hiccups pre-treatment and post-treatment. Adverse events were also recorded. RESULTS: Absolute resolution was observed in all 15 patients after less than three acupuncture sessions. Of these, 10 patients required only one acupuncture session, 3 patients required two sessions and 2 patients required three sessions. The HAI score improved after each round of acupuncture treatment (P<0.05). The average HAI score improved significantly post-acupuncture compared to baseline values pre-treatment (P<0.05). Symptoms accompanying the hiccups included pain in the diaphragmatic area (five patients), mild dyspnoea (three patients), dysphagia (two patients) and nausea/vomiting (one patient). All these accompanying symptoms disappeared at the point of resolution of the hiccups. There were no adverse effects related to acupuncture during the study period. CONCLUSION: Based on our results, acupuncture may represent a potential treatment option for hiccups after arthroplasty. Caution must be exercised, however, given the lack of a control group. Accordingly, randomised controlled trials will be required to verify the efficacy and effectiveness of acupuncture for the treatment of hiccups.

Effect of ketorolac in intra-articular injection analgesia for postoperative pain in patients undergoing shoulder arthroscopy: a pilot-controlled clinical study.

BACKGROUND: To date, a regional approach using local anesthetics has become a popular analgesic method for arthroscopy. The optimal postoperative analgesia method for shoulder arthroscopy is still debated. OBJECTIVE: This study was designed to evaluate the effect and safety of using ketorolac in combination with a multimodal drug regime (ropivacaine, morphine, and triamcinolone acetonide) after shoulder arthroscopy. METHODS: A total of 60 patients were included in a pilot study and patients were randomized into an experimental group (n=30) and a control group (n=30). The following parameters were used to evaluate pain relief levels postoperatively: the Visual Analog Scale (VAS) at 1, 3, 6, 12, 24, and 48 hours postoperatively, morphine consumption, and initial analgesic desired time. Complications were also recorded. RESULTS: Except for 1 hour postoperatively, patients in the experimental group experienced lower VAS scores during the first 48 hours postoperatively (P<0.05). The VAS score in both groups increased after 3 hours postoperatively and peaked at 12 hours postoperatively (2.54±0.86 vs 3.25±1.18). The VAS scores on movement in the experimental group were lower than those in the control group at 24 or 48 hours postoperatively (P=0.004, 0.001). A total of 18 (60.0%) patients in the experimental group required no additional analgesia, compared with 10 (33.3 %) in the control group (P=0.035). The mean rescue analgesia was 11.40±5.56 mg in the experiment group, while 16.57±8.48 mg in the control group (P=0.016). The initial analgesic desired time was delayed significantly in the experimental group (16.50±14.57 hours vs 8.9±6.32 hours, P=0.000). CONCLUSION: Adding ketorolac to intra-articular injection analgesia is a safe and effective method to improve pain relief after shoulder arthroscopy, and further prospective controlled trials are necessary to allow definite treatment recommendations.

The severity of ulnar variance compared with contralateral hand: its significance on postoperative wrist function in patients with distal radius fracture.

The aim of this study was to detect the severity of ulnar variance (UV) compared with contralateral hand on postoperative wrist function in patients with distal radius fracture. 116 cases with unilateral distal radius fracture were retrospectively analyzed and divided into high or low UV severity groups (Dividing value = 2.5 mm). The following parameters were used to evaluate the effect: palmar tilt, radial inclination, VAS score, DASH score and wrist function. The severity of UV existed widely, accounting for 93.1% (108 cases). The severity of UV correlated with palmar tilt, radial inclination, grip strength, VAS score, DASH score and the wrist function (P < 0.05). Log-rank analysis showed that the severity of UV, palmar tilt, radial inclination were important factors influencing the joint function postoperatively (P < 0.0 5). Multivariate analysis confirmed that the severity of UV was an independent and significant factor on wrist function (P = 0.010). And the palmar tilt was also an important factor influencing wrist function (P = 0.047). The severity of ulnar variance compared with contralateral hand is an independent and significant factor on wrist function, which should be considered as an important step during preoperative plan.

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.

The efficacy of pre-emptive analgesia on pain management in total knee arthroplasty: a mini-review.

Total knee arthroplasty (TKA) is considered a cost-effective and efficacious treatment for patients with end-stage knee arthritis. Meanwhile, TKA has been regarded as one of the most painful orthopaedic surgeries. Pain control after TKA remains a challenging task. Many analgesic innovations are used to reduce the level of pain, but none has been proven to be the optimum choice till now. Multimodal analgesia incorporates the use of analgesic adjuncts with different mechanisms of action to enhance postoperative pain management. This approach is a preferable choice in relieving postoperative pain with minimum side effects. This paper aims to review pre-emptive analgesia for pain management in TKA. We reviewed the application of pre-emptive analgesia, its physiological mechanism, and the techniques.

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.