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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. [ABSTRACT FROM AUTHOR]

BACKGROUND: Previous studies on the clinical efficacy of total knee arthroplasty in the treatment of rheumatoid arthritis have been reported, but the effects of total knee arthroplasty on joint function and related biochemical parameters in patients with rheumatoid arthritis are little reported. Thereafter, more clinical evidence is needed. OBJECTIVE: To analyze the effect of total knee arthroplasty on joint function and related biochemical indicators in patients with rheumatoid arthritis. METHODS: Clinical data of 64 patients with 64 knees who underwent total knee arthroplasty were retrospectively analyzed. Comparative analysis of Hospital for Special Surgery, range of motion of the knee joint and quality of life scores before and 1 year after surgery was conducted. Changes of biochemical indicators such as C-reactive protein, rheumatoid factor and erythrocyte sedimentation rate were detected. The incidence of complications at 1 month after surgery was recorded. The study was approved by the Ethical Committee of Changzhou Traditional Chinese Medicine Hospital, and all patients signed the informed consents. RESULTS AND CONCLUSION: (1) Compared with the baseline levels, the Hospital for Special Surgery score, range of motion of the knee joint and quality of life scores at 1 year after surgery was significantly increased (P < 0.01 ), and the levels of C-reactive protein, rheumatoid factor and erythrocyte sedimentation rate were significantly decreased (P < 0.01 ). (2) One patient developed infection and two patients developed deep vein thrombosis of the lower extremity, which was relieved after symptomatic treatment. These results imply that total knee arthroplasty has a good dinical effect on rheumatoid arthritis, which can improve knee function and reduce the levels of C-reactive protein, rheumatoid factor and erythrocyte sedimentation rate, and has good safety. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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.

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.

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).

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 (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.

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.