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Wen-Jiao Wu,1,* Chang-Liang Xia,1,* Shuan-Ji Ou,1 Yang Yang,1 Xiao-Zhong Zhou,1 Yun-Fei Ma,2 Yi-Long Hou,2 Fa-Zheng Wang,3 Qing-Po Yang,3 Yong Qi,1,4 Chang-Peng Xu1,4 1Department of Orthopaedics, Guangdong Second Provincial General Hospital, Guangzhou, Guangdong, People’s Republic of China; 2Department of Orthopaedics and Traumatology, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, People’s Republic of China; 3Department of Orthopaedics, First People’s Hospital of Kashgar Prefecture, Kashgar, Xinjiang, People’s Republic of China; 4The Second School of Clinical Medicine, Southern Medical University, Guangzhou, Guangdong, People’s Republic of China*These authors contributed equally to this workCorrespondence: Yong Qi; Chang-Peng Xu, Tel +86-20-8916-8085, Email gd2hqy@163.com; gd2hxcp@163.comIntroduction: Osteomyelitis is characterized by intensive inflammatory bone disease and remains a clinical challenge in orthopedic surgery, despite the advances made in medical and surgical therapies. Staphylococcus aureusis a major causative agent of osteomyelitis, causing the progressive inflammatory destruction of bone. Prophylaxis of osteomyelitis during orthopedic surgery is necessary. NFκB essential modulator–binding domain (NBD) peptides are cell-permeable peptide inhibitors of the IκB-kinase complex. The prophylactic effect of NBD peptides in relieving inflammation and inhibiting bone defects in osteomyelitis is still under investigation. Our purpose was to determine the preventive effect of NBD peptides in S. aureus infection–induced bone defects in osteomyelitis.Methods: An S. aureus osteomyelitis rabbit model was used in this study. The rabbits were divided into four groups: NBD, cefazolin, control, and PBS. Clinical and laboratory indicators of erythrocyte-sedimentation rate, CRP, and TNFα levels were assessed to monitor systemic reactions. The efficacy of NBD peptides in S. aureus–induced osteomyelitis was evaluated by radiological, histological, and microbiological examinations, immunohistochemistry, immunofluorescence, and micro-CT scans.Results: In general, NBD peptides effectively reduced clinical signs in rabbits when compared with the control group. Radiography indicated that there was more severe osteomyelitis in the bacterium-infection control group. There was no significance between cefazolin- and NBD-group average scores. The histological results of the lesion slices further confirmed different severity among the groups. Additionally, significant pathological differences were found between the cefazolin and NBD groups, and the PBS group showed no obvious pathological changes.Conclusion: Prophylactic administration of NBD peptides to bone-defect areas inhibited bacterial spread and promoted bone regeneration, making NBD peptides a possible treatment option for prophylaxis in bone infections.Keywords: NBD peptide, bone infection, Staphylococcus aureus, osteomyelitis

Sustained activation of multiple receptor tyrosine kinases (RTKs) simultaneously is vital for tumorigenesis and progression of osteosarcoma (OS). Gαi proteins recruitment to various RTKs mediates downstream oncogenic signaling activation. The expression, functions and underlying mechanisms of Gαi3 in human OS were examined. Expression of Gαi3 is robustly elevated in human OS tissues and is correlated with a poor overall survival. In patient-derived primary OS cells and immortalized lines (MG63 and U2OS), Gαi3 depletion, by shRNA and CRISPR/Cas9 strategies, robustly suppressed cell viability, proliferation and migration, while provoking G1-S arrest and apoptosis activation. Conversely, Gαi3 overexpressing ectopically can accelerate proliferation and migration of OS cells. In OS cells, Gαi3 immunoprecipitated with VEGFR2, FGFR, PGDFR and EGFR, mediating downstream cascade transduction. Akt-mTOR activation in primary OS cells was potently inhibited by Gαi3 shRNA, knockout or dominant negative mutation, but augmented after Gαi3 overexpression.

BACKGROUND: Cartilage tissue lacks the ability to heal. Cartilage tissue engineering using cell-free scaffolds has been increasingly used in recent years. OBJECTIVE: This study describes the use of a type I collagen scaffold combined with WNT5A plasmid to promote chondrocyte proliferation and differentiation in a rabbit osteochondral defect model. METHODS: Type I collagen was extracted and fabricated into a collagen scaffold. To improve gene transfection efficiency, a cationic chitosan derivative N,N,N-trimethyl chitosan chloride (TMC) vector was used. A solution of TMC/WNT5A complexes was adsorbed to the collagen scaffold to prepare a WNT5A scaffold. Osteochondral defects were created in the femoral condyles of rabbits. The rabbits were divided into defect, scaffold, and scaffold with WNT5A groups. At 6 and 12 weeks after creation of the osteochondral defects, samples were collected from all groups for macroscopic observation and gene expression analysis. RESULTS: Samples from the defect group exhibited incomplete cartilage repair, while those from the scaffold and scaffold with WNT5A groups exhibited “preliminary cartilage” covering the defect. Cartilage regeneration was superior in the scaffold with WNT5A group compared to the scaffold group. Safranin O staining revealed more proteoglycans in the scaffold and scaffold with WNT5A groups compared to the defect group. The expression levels of aggrecan, collagen type II, and SOX9 genes were significantly higher in the scaffold with WNT5A group compared to the other two groups. CONCLUSIONS: Type I collagen scaffold showed effective adsorption and guided the three-dimensional arrangement of stem cells. WNT5A plasmid promoted cartilage repair by stimulating the expression of aggrecan, type II collagen, and SOX9 genes and proteins, as well as inhibiting cartilage hypertrophy.

Objective To evaluate the effectiveness and safety of allogeneic tendons for functional reconstruction of severe hand injuries. Methods From August 2007 to July 2014, we performed functional reconstruction with tendon allografts for severe hand injuries affecting two or more tendons. At the final follow-up, we assessed total active motion (TAM); pincer pinch strength; grip strength; Disabilities of the Arm, Shoulder, and Hand (DASH) score; degree of satisfaction; and adhesion. We measured the white blood cell count, C-reactive protein concentration, erythrocyte sedimentation rate, total T-cell count, and CD4+T/CD8+T ratio to evaluate the immune response and check for infection. Results Ten patients received 26 allogeneic tendons to reconstruct hand function. The average follow-up period was 50.0 months (range, 24–82 months). The TAM was 126.4° (12°–253°), pincer pinch strength was 0.83 kg (0–4.5 kg), and grip strength was 13.69 kg (4–41.5 kg). The DASH score was 14.25 (3.3–30.8), and seven and three patients were satisfied and partially satisfied, respectively. One patient developed tendon adhesion. All immune and infectious parameters were within the reference range. Conclusion Functional reconstruction using allogeneic tendons for severe hand injuries with multiple tendon defects was effective and safe; however, more research is needed.

Osteosarcoma (OS) is the most common primary bone malignancy in the adolescent population. Recent studies demonstrate that p38 gamma (p38γ) phosphorylates retinoblastoma (Rb) to promote cyclin expression, cell-cycle entry and tumorigenesis. Studying the potential function of p38γ in human OS, we show that p38γ mRNA and protein expression are significantly elevated in OS tissues and OS cells, whereas its expression is relatively low in normal bone tissue and in human osteoblasts/osteoblastic cells. Knockdown of p38γ in established (U2OS) and primary human OS cells potently inhibited cell growth, proliferation, migration and invasion, while promoting cell apoptosis. Furthermore, CRISPR/Cas9-induced p38γ knockout inhibited human OS cell progression in vitro. Conversely, ectopic overexpression of p38γ in primary human OS cells augmented cell growth, proliferation and migration. Signaling studies show that retinoblastoma (Rb) phosphorylation and cyclin E1/cyclin A expression were decreased following p38γ shRNA knockdown and knockout, but increased after ectopic p38γ overexpression. Collectively, these results show that p38γ overexpression promotes human OS cell progression.

The present study determines the role of the Gab1 in hydrogen peroxide (H2O2)-induced death of human osteoblasts. We show that Gab1 is required for H2O2-induced Akt activation to promote osteoblast survival. In OB-6 human osteoblasts, Gab1 silencing (by targeted-shRNA) or complete knockout (by CRISPR-Cas9 KO plasmid) largely attenuated Akt activation by H2O2. Gab1-depleted OB-6 cells were more vulnerable to H2O2. Conversely, forced over-expression of Gab1 by an adenovirus vector increased Akt activation to protect OB-6 cells from H2O2. Significantly, the anti-sense of microRNA-29a (“antagomiR-29a”) induced Gab1 expression to facilitate H2O2-induced Akt activation, which protected OB-6 cells from apoptosis. AntagomiR-29a was however ineffective in Gab1-deficient and Akt-inhibited OB-6 cells. Forced over-expression of miR-29a induced Gab1 downregulation to inhibit H2O2-induced Akt activation, causing enhanced OB-6 cell death. miR-29a-induced actions were abolished by an adenovirus constitutively-active Akt1 (Ad-caAkt1) in OB-6 cells. Together, microRNA-29a inhibition induces Gab1 upregulation and Akt activation to protect OB-6 osteoblasts from H2O2.

The present study examined the expression and biological functions of bromodomain-containing protein 4 (BRD4) in skin squamous cell carcinoma (SCC) cells. Our results show that BRD4 mRNA and protein expression was upregulated in human skin SCC cells, as compared to its level in the normal skin keratinocytes and fibroblasts. Treatment with BRD4 inhibitors, JQ1 and CPI203, resulted in proliferation inhibition, apoptosis and cell cycle arrest in both established (A431 cell line) and primary skin SCC cells. Furthermore, BRD4 knockdown (by targeted shRNAs) or knockout (by CRISPR/Cas9) largely inhibited A431 cell proliferation. Reversely, forced-overexpression of BRD4 in A431 cells facilitated cell proliferation. We show that BRD4 is required for the expression of several oncogenes, including cyclin D1, Bcl-2 and MYC. BRD4 inhibition, knockdown or knockout significantly decreased above oncogene expression in SCC cells. In vivo, CRISPR/Cas9-mediated BRD4 knockout significantly suppressed A431 xenograft tumor growth in severe combined immunodeficient (SCID) mice. Together, our results suggest that BRD4 could be a novel and pivotal oncogenic protein of skin SCC.

// Shang Cai 1, * , Ya Li 2, * , Jin-Yu Bai 3, * , Zhi-Qing Zhang 2, * , Yin Wang 2 , Yin-Biao Qiao 4 , Xiao-Zhong Zhou 3 , Bo Yang 4 , Ye Tian 1 , Cong Cao 2 1 Department of Radiotherapy and Oncology, The Second Affiliated Hospital of Soochow University, Suzhou, China 2 Institute of Neuroscience, Soochow University, Suzhou, China 3 Department of Orthopedics, The Second Affiliated Hospital of Soochow University, Suzhou, China 4 Department of Surgery, The Third Hospital affiliated to Soochow University, Suzhou, China * Co-first authors Correspondence to: Bo Yang, email: dryangbosz@163.com Ye Tian, email: tianyesuzhoua163@163.com , dryetian@126.com Cong Cao, email: caocong@suda.edu.cn Keywords: glioma, irradiation, Gαi3, DNA-PKcs Received: March 13, 2017 Accepted: March 30, 2017 Published: April 11, 2017 ABSTRACT We have previously shown that Gαi3 is elevated in human glioma, mediating Akt activation and cancer cell proliferation. Here, we imply that Gαi3 could also be important for irradiation resistance. In A172 human glioma cells, Gαi3 knockdown (by targeted shRNAs) or dominant-negative mutation significantly potentiated irradiation-induced cell apoptosis. Reversely, forced over-expression of wild-type or constitutively-active Gαi3 inhibited irradiation-induced A172 cell apoptosis. Irradiation in A172 cells induced Gαi3 translocation to cell nuclei and association with local protein DNA-dependent protein kinase (DNA-PK) catalytic subunit. This association was important for DNA damage repair. Gαi3 knockdown, depletion (using Gαi3 knockout MEFs) or dominant-negative mutation potentiated irradiation-induced DNA damages. On the other hand, expression of the constitutively-active Gαi3 in A172 cells inhibited DNA damage by irradiation. Together, these results indicate a novel function of Gαi3 in irradiation-resistance in human glioma cells.

// Yun-fang Zhen 1, * , Song-tao Li 2, * , Yun-rong Zhu 3, * , Xiao-dong Wang 1 , Xiao-zhong Zhou 2 , Lun-qing Zhu 1 1 The Center of Diagnosis and Treatment for Children’s Bone Diseases, The Children’s Hospital Affiliated to Soochow University, Suzhou, China 2 The Department of Orthopedics, The Second Affiliated Hospital of Soochow University, Suzhou, China 3 Department of Orthopedics, The Affiliated Jiangyin Hospital of Medical College of Southeast University, Jiangyin, China * These authors have contributed equally to this work Correspondence to: Lun-qing Zhu, email: zhulunqingszgk@163.com Xiao-dong Wang, email: xiaodongwangsz@163.com Xiao-zhong Zhou, email: zhouxiaozhongorth@163.com Keywords: osteosarcoma (OS), salinomycin, DNA-PKcs, microRNA-101, autophagy Received: September 09, 2016 Accepted: September 26, 2016 Published: October 17, 2016 ABSTRACT Malignant osteosarcoma (OS) is still a deadly disease for many affected patients. The search for the novel anti-OS agent is extremely urgent and important. Our previous study has proposed that salinomycin is a novel anti-OS agent. Here we characterized DNA-dependent protein kinase catalytic subunit (DNA-PKcs) as a primary salinomycin resistance factor in OS cells. DNA-PKcs inhibitors (NU7026, NU7441 and LY294002) or DNA-PKcs shRNA knockdown dramatically potentiated salinomycin-induced death and apoptosis of OS cells (U2OS and MG-63 lines). Further, forced-expression of microRNA-101 (“miR-101”) downregulated DNA-PKcs and augmented salinomycin’s cytotoxicity against OS cells. Reversely, over-expression of DNA-PKcs in OS cells inhibited salinomycin’s lethality. For the mechanism study, we show that DNA-PKcs is required for salinomycin-induced pro-survival autophagy activation. DNA-PKcs inhibition (by NU7441), shRNA knockdown or miR-101 expression inhibited salinomycin-induced Beclin-1 expression and autophagy induction. Meanwhile, knockdown of Beclin-1 by shRNA significantly sensitized salinomycin-induced OS cell lethality. In vivo , salinomycin administration suppressed U2OS xenograft tumor growth in severe combined immuno-deficient (SCID) mice, and its anti-tumor activity was dramatically potentiated with co-administration of the DNA-PKcs inhibitor NU7026. Together, these results suggest that DNA-PKcs could be a primary resistance factor of salinomycin in OS cells. DNA-PKcs inhibition or silence may thus significantly increase salinomycin’s sensitivity in OS cells.

// Min-Bin Chen 1, * , Zhen-Tao Zhou 2, * , Lan Yang 3, * , Mu-Xin Wei 4 , Min Tang 1 , Ting-Yan Ruan 5 , Jun-Ying Xu 5 , Xiao-zhong Zhou 2 , Gang Chen 6 , Pei-Hua Lu 5 1 Department of Oncology, Kunshan First People’s Hospital Affiliated to Jiangsu University, Kunshan 215300, China 2 Department of Orthopedics, the Second Affiliated Hospital of Soochow University, Suzhou 215000, China 3 Department of Breast Surgery, the Third Affiliated Hospital of Soochow University, Changzhou 213003, China 4 Department of Traditional Chinese Medicine, First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China 5 Department of Medical Oncology, Wuxi People’s Hospital Affiliated to Nanjing Medical University, Wuxi 214023, China 6 Department of Neurosurgery, the First Affiliated Hospital of Soochow University, Suzhou 215006, China * These authors have contributed equally to this work Correspondence to: Pei-Hua Lu, e-mail: lphty1_1@163.com Xiao-zhong Zhou, e-mail: zhouxz@suda.edu.cn Gang Chen, e-mail: nju_neurosurgery@163.com Keywords: hepatocellular carcinoma (HCC), DNA-PKcs, KU-0060648, PI3K-AKT-mTOR signaling, miRNA Received: October 22, 2015 Accepted: February 05, 2016 Published: February 26, 2016 ABSTRACT Here we tested anti-tumor activity of KU-0060648 in preclinical hepatocellular carcinoma (HCC) models. Our results demonstrated that KU-0060648 was anti-proliferative and pro-apoptotic in established (HepG2, Huh-7 and KYN-2 lines) and primary human HCC cells, but was non-cytotoxic to non-cancerous HL-7702 hepatocytes. DNA-PKcs (DNA-activated protein kinase catalytic subunit) is an important but not exclusive target of KU-0060648. DNA-PKcs knockdown or dominant negative mutation inhibited HCC cell proliferation. On the other hand, overexpression of wild-type DNA-PKcs enhanced HepG2 cell proliferation. Importantly, KU-0060648 was still cytotoxic to DNA-PKcs-silenced or -mutated HepG2 cells, although its activity in these cells was relatively weak. Further studies showed that KU-0060648 inhibited PI3K-AKT-mTOR activation, independent of DNA-PKcs. Introduction of constitutively-active AKT1 (CA-AKT1) restored AKT-mTOR activation after KU-0060648 treatment in HepG2 cells, and alleviated subsequent cytotoxicity. In vivo , intraperitoneal ( i.p. ) injection of KU-0060648 significantly inhibited HepG2 xenograft growth in nude mice. AKT-mTOR activation was also inhibited in xenografted tumors. Finally, we showed that DNA-PKcs expression was significantly upregulated in human HCC tissues. Yet miRNA-101, an anti-DNA-PKcs miRNA, was downregulated. Over-expression of miR-101 in HepG2 cells inhibited DNA-PKcs expression and cell proliferation. Together, these results indicate that KU-0060648 inhibits HCC cells through DNA-PKcs-dependent and -independent mechanisms.

Fil: Marshall, John. Brown University. Department of Molecular Pharmacology, Physiology, and Biotechnology; Estados Unidos Fil: Zhou, Xiao-Zhong. Soochow University. Jiangsu Key Laboratory of Neuropsychiatric Diseases Research; China Fil: Zhou, Xiao-Zhong. Soochow University. Institute of Neuroscience; China Fil: Zhou, Xiao-Zhong. Second Affiliated Hospital of Soochow University. Department of Orthopedics; China Fil: Chen, Gang. First Affiliated Hospital of Soochow University. Department of Neurosurgery; China Fil: Yang, Su-Qing. Soochow University. Jiangsu Key Laboratory of Neuropsychiatric Diseases Research; China Fil: Yang, Su-Qing. Soochow University. Institute of Neuroscience; China Fil: Li, Ya. Jiangsu Key Laboratory of Neuropsychiatric Diseases Research; China Fil: Li, Ya. Soochow University. Institute of Neuroscience; China Fil: Wang, Yin. Jiangsu Key Laboratory of Neuropsychiatric Diseases Research; China Fil: Wang, Yin. Soochow University. Institute of Neuroscience; China Fil:Zhang, Zhi-Qing. Jiangsu Key Laboratory of Neuropsychiatric Diseases Research; China Fil:Zhang, Zhi-Qing. Soochow University. Institute of Neuroscience; China Fil: Jiang, Qin. Nanjing Medical University. The Affiliated Eye Hospital. The Fourth School of Clinical Medicine; China Fil: Birnbaumer, Lutz. National Institute of Environmental Health Sciences. Neurobiology Laboratory; Estados Unidos Fil: Birnbaumer, Lutz. Pontificia Universidad Católica Argentina. Facultad de Ciencias Médicas. Instituto de Investigaciones Biomédicas; Argentina Fil: Cao, Cong. Jiangsu Key Laboratory of Neuropsychiatric Diseases Research; China Fil: Cao, Cong. Soochow University. Institute of Neuroscience; China Fil: Cao, Cong. Nanjing Medical University. The Affiliated Eye Hospital. The Fourth School of Clinical Medicine; China Fil: Cao, Cong. The Municipal Hospital of Suzhou. North District; China Abstract: Stress-related alterations in brain-derived neurotrophic factor (BDNF) expression, a neurotrophin that plays a key role in synaptic plasticity, are believed to contribute to the pathophysiology of depression. Here, we show that in a chronic mild stress (CMS) model of depression the Gαi1 and Gαi3 subunits of heterotrimeric G proteins are down-regulated in the hippocampus, a key limbic structure associated with major depressive disorder. We provide evidence that Gαi1 and Gαi3 (Gαi1/3) are required for the activation of TrkB downstream signaling pathways. In mouse embryonic fibroblasts (MEFs) and CNS neurons, Gαi1/3 knockdown inhibited BDNF-induced tropomyosin-related kinase B (TrkB) endocytosis, adaptor protein activation, and Akt-mTORC1 and Erk-MAPK signaling. Functional studies show that Gαi1 and Gαi3 knockdown decreases the number of dendrites and dendritic spines in hippocampal neurons. In vivo, hippocampal Gαi1/3 knockdown after bilateral microinjection of lentiviral constructs containing Gαi1 and Gαi3 shRNA elicited depressive behaviors. Critically, exogenous expression of Gαi3 in the hippocampus reversed depressive behaviors in CMS mice. Similar results were observed in Gαi1/Gαi3 double-knockout mice, which exhibited severe depressive behaviors. These results demonstrate that heterotrimeric Gαi1 and Gαi3 proteins are essential for TrkB signaling and that disruption of Gαi1 or Gαi3 function could contribute to depressive behaviors.

The brush type plasticizer system in KDF2 filter rod maker tends to result in wide variation in triacetin addition and material waste. In order to improve the quality of filter rod, an automatic triacetin spraying system based on an atomizing technology was designed, wherein PLC and touch screen acted as main controllers. Heated triacetin was pumped into a manifold under high-pressure then evenly sprayed on the running tow by nozzle. By means of the built-in positioning function of PLC and servo positioning module, the tow speed was tracked, the plasticizer valve was accurately controlled to adjust the flow rate of triacetin. By detecting the triacetin temperature and level in the plasticizer booth, triacetin temperature and volume were automatically maintained. The practical testing results showed that the filter rod hardness greater than or equal to 80%. Triacetin was sprayed evenly and the system ran smoothly and reliably.

// Zheng-Wei Li 1, * , Yun-Rong Zhu 2, * , Xiao-Zhong Zhou 3, 4, * , Bao-Biao Zhuo 1 , Xiao-Dong Wang 1 1 The Center of Diagnosis and Treatment for Children’s Bone Diseases, The Children’s Hospital Affiliated to Soochow University, Suzhou, China 2 Department of Orthopedics, The Affiliated Jiangyin Hospital of Medical College of Southeast University, Jiangyin City, China 3 Department of Orthopedics, The Second Affiliated Hospital of Soochow University, Suzhou, China 4 Department of Orthopedics, The First People’s Hospital of SuQian, SuQian, China * Co-first authors Correspondence to: Xiao-Dong Wang, email: xiaodongwangsz@163.com Keywords: microRNA-135b, Ppm1e, AMPK, osteoblastoma, cell proliferation Received: January 20, 2017 Accepted: February 08, 2017 Published: February 18, 2017 ABSTRACT Forced-activation of AMP-activated protein kinase (AMPK) can possibly inhibit osteoblastoma cells. Here, we aim to provoke AMPK activation via microRNA silencing its phosphatase Ppm1e (protein phosphatase Mg 2+ /Mn 2+ -dependent 1e). We showed that microRNA-135b-5p (“miR-135b-5p”), the anti-Ppm1e microRNA, was significantly downregulated in human osteoblastoma tissues. It was correlated with Ppm1e upregulation and AMPKα1 de-phosphorylation. Forced-expression of miR-135b-5p in human osteoblastoma cells (MG-63 and U2OS lines) silenced Ppm1e, and induced a profound AMPKα1 phosphorylation (at Thr-172). Osteoblastoma cell proliferation was inhibited after miR-135b-5p expression. Intriguingly, Ppm1e shRNA knockdown similarly induced AMPKα1 phosphorylation, causing osteoblastoma cell proliferation. Reversely, AMPKα1 shRNA knockdown or dominant negative mutation almost abolished miR-135b-5p’s actions in osteoblastoma cells. Further in vivo studies demonstrated that U2OS tumor growth in mice was dramatically inhibited after expressing miR-135b-5p or Ppm1e shRNA. Together, our results suggest that miR-135b-induced Ppm1e silence induces AMPK activation to inhibit osteoblastoma cell proliferation.

With the introduction and wide applications of laser guided AGV in the tobacco corporation, the logistics automation level is increased greatly. But with the introduction of flexible manufacturing system, the production line of the tobacco corporation become more complicate, the material requirement become more flexible, so that the driving stability has brought great challenges of AGV. In the system of more AGV compositing, there will been bought the conflict between multiple vehicles AGV. How can locate the position of each AGV instantly, and estimate and dispose the conflict during the running process in time, it has been the solve direction for team. This paper is researched based on the AGV’s system of KunChuan, bring forward a monitoring system of AGVS. The results shown that the actual trajectory is fully consistent with the planned through running, and achieving the prospective effect.

Prolonged or overdose glucocorticoids (GCs) usage is the common cause of osteoporosis. In the present study, we studied the cellular mechanism of dexamethasone (Dex)-induce osteoblast cell death by focusing on the role of mitochondrial permeability transition pore (mPTP). In cultured osteoblastic MC3T3-E1 cells, Dex-induced mPTP opening, which was demonstrated by mitochondrial membrane potential (MPP) decrease, cyclophilin-D (CyPD)-adenine nucleotide translocator 1 (ANT-1) mitochondrial complexation and cytochrome C (cyto-C) release. The mPTP inhibitor sanglifehrin A (SfA) dramatically inhibited Dex-induced MPP loss, cyto-C release and MC3T3-E1 cell death. Dex-induced cell death requires mPTP composing protein CyPD, as CyPD inhibitor cyclosporin A (CsA) and CyPD siRNA knockdown inhibited Dex-induced MC3T3-E1 cell death, while CyPD overexpression aggravated Dex's cytotoxic effect. We found that Dex induced P53 phosphorylation and translocation to mitochondria, where it formed a complex with CyPD. Glucocorticoid receptor (GR) siRNA knockdown, or P53 inhibition (by its inhibitor pifithrin-α or shRNA silencing) suppressed Dex-induced CyPD-P53 mitochondrial association and subsequent MC3T3-E1 cell death. Finally, in primary cultured osteoblasts, Dex-induced cell death was inhibited by CsA, SfA or pifithrin-α. Together, our data suggest that Dex-induced osteoblast cell death is associated with GR-P53-regulated mPTP opening.

The cut tobacco transporting is the absolutely necessarily important tache, whether the transportation process stability is a direct impact on production quality and manufacturing standards. The wind system is adopted by most tobacco companies because of it caused least disruptive of the cut tobacco structure and the pipe network layout of system is flexible, but the wind system is greater influence by itself, different control methods are caused greater difference of process indicators, for example essence and spice ingredients, moisture proportion and smoke flavor. Based on the above reasons, so we need to design the control method of wind system necessarily, in addition we must checkout the control accuracy opportunely, so that it is satisfied by the actual working conditions.

Dexamethasone (Dex) causes osteoblast cell injuries. In the present research, we tested the potential effect of SC79, a novel and specific Akt activator, against Dex in osteoblasts. In primary murine osteoblasts and osteoblastic MC3T3-E1 cells, pretreatment with SC79 significantly attenuated Dex-induced cell death. Further, Dex-induced mitochondrial permeability transition pore (mPTP) opening, cytochrome C release and apoptosis activation were dramatically alleviated with SC79 pretreatment in above cells. At the molecular level, SC79 activated Akt, which was indispensable for subsequent osteoblast protection against Dex. Akt inhibitors (LY294002, perifosine and MK-2206) blocked SC79-induced Akt activation and abolished its anti-Dex actions in osteoblasts. Further, SC79 activated Akt downstream Nrf2 (NF-E2-related factor 2) signaling and attenuated Dex-induced oxidative stress in osteoblasts. Nrf2 shRNA knockdown or S40T mutation almost reversed SC79-mediated anti-oxidant and cytoprotective activities in osteoblasts. Together, these results suggest that SC79 activates Akt-Nrf2 signaling to protect osteoblasts from Dex.

A novel Cu–10Ni–20Zn–1.6Al (wt.%) alloy with high strength through precipitation hardening is reported in this paper. The yield strength and ultimate tensile strength of this alloy can reach maximum peaks of 1034 MPa and 1112 MPa, respectively. The nanosized coherent precipitates with the Ll2-type ordered structure, which result in precipitation hardening, are formed homogenously in the matrix during the isothermal ageing, and the precipitates can produce zones of severe distortion in the matrix surrounding them due to the coherency.

The effect of aluminum on the precipitation hardening of Cu–Ni–Zn alloys with varying aging temperatures and times was investigated in this article, in the hope to achieve better mechanical properties. Vickers hardness, tensile, and electrical conductivity tests were carried out to characterize the properties of the Cu–Ni–Zn alloys with or without an addition of aluminum subjected to different aging treatments. The results show that an addition of 1.2 wt% aluminum can play an influential role in the precipitation hardening of the Cu–Ni–Zn alloys. For example, it can increase the peak hardness from 58 Hv for the solution-treated Cu–10Ni–20Zn alloy to 185 Hv for the solution-treated Cu–10Ni–20Zn–1.2Al alloy during aging at 500 °C. The yield strength, tensile strength, and electrical conductivity of the Cu–10Ni–20Zn–1.2Al alloy subjected to suitable treatments under prior cold-rolled and aged conditions can reach 889 MPa, 918 MPa, and 10.96% IACS, respectively, being much higher than those of the relevant alloy without aluminum and comparable to those of the Cu–Be alloys (C17200 and C17510). According to the transmission electron microscope observations, it was found that formation of nanosized precipitates with the L12-type ordered lattice results in precipitation hardening, and an orientation relationship of $$ [011]_{\text{p}}//[011]_{\text{m}} $$ and $$ (100)_{\text{p}}//(200)_{\text{m}} $$ exists between the precipitates and the α-Cu matrix.

The electrochemical performance of the Cu–20Zn–10Ni alloy was compared with that of the Cu–30Ni alloy in a 3.5 wt.% sodium chloride solution. Electrochemical impedance spectroscopy (EIS) measurements show that, interestingly, both the increasing rates of the thickness and resistance of the passive film formed on the Cu–20Zn–10Ni alloy are larger than those of the passive film formed on the Cu–30Ni alloy during the immersion of the two alloys in the 3.5 wt.% NaCl solution. The passive film formed on the Cu–20Zn–10Ni alloy surface is found to have a higher resistance and thicker depth than that formed on the Cu–30Ni alloy surface after the immersion of the two alloys in the 3.5 wt.% NaCl solution for 120 h. To some extent, the Cu–20Zn–10Ni alloy could substitute for the Cu–30Ni alloy in a corrosive environment containing chloride ions. According to X-ray diffraction and scanning electron microscopy studies, the corrosion product films of the Cu–20Zn–10Ni alloy were found to consist of two layers, with the inner layer and the outer layer being composed of cuprous oxide and (Cu, Zn) 2 Cl(OH) 3 , respectively, while the corrosion product films of the Cu–30Ni alloy were found to consist mainly of (Cu, Ni) 2 Cl(OH) 3 .

The formation and evolution of the solid electrolyte interface (SEI) film during the first electrochemical intercalation of lithium into graphite were modeled as a special precipitation process including a nucleation phase of the SEI film's solid deposition, and followed by a growth phase involving the precipitation of new solids on previously formed solid nuclei. It was shown that the solid species can nucleate in the electrolyte solution, directly on the graphite surface, or adjacent to an already present particle on the graphite surface when precipitating from the electrolyte solution. Within the framework of classical nucleation theory (CNT), we can qualitatively explain the origin of the two-layer structure of SEI films, which consists of a thin, compact polycrystalline or heteromicrophase layer rich in inorganic species ( e.g. , LiF, Li 2 O) close to the electrode, and a thicker porous and amorphous layer composed mainly of organic compounds ( e.g. , ROLi, ROCO 2 Li) that is farther from the graphite.

Low-dose irradiation (LDI) exhibits a positive effect on osteoblasts and inhibitory effect of inflammation. Here, we test the hypothesis that LDI can promote osseointegration and inhibit the inflammatory membrane formation in the presence of titanium (Ti) particles. Endotoxin-free titanium particles were injected into rabbit, prior to the insertion of a Ti6-Al-4-V sticks pre-coated with hydroxyapatite. Two days after operation, both distal femurs of the animal were exposed to 0.5 Gy X-ray irradiation. All ani-mals were euthanized 8 weeks after the operation. The PINP concentration was determined at day 0, 2, 4, and 8 weeks after operation. Trabecular morphology around the implants 8 weeks after operation was assessed using micro-CT, then the maximum push out force of simples was assessed using biomechanics test. Five samples in each group were chosen for bone histomorphology study without decalcification 8 weeks after operation. The results confirmed that the LDI can significantly improve ingrowth of bone into the prosthetic interface and stability of the prosthesis when there was no wear particles. Although promotion effects for bone formation induced by LDI can be counteracted by wear particles, LDI can significantly inhibit the interface membrane formation around the implant induced by wear particles. Based on these results, we conclude that LDI may be useful for enhancing the stability of prosthesis when there are no wear particles and for inhibiting the interface membrane formation during the early stage of aseptic loosening in the presence of wear particles. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 34:1688-1696, 2016.

Low-dose irradiation (LDI) induces osteoblast differentiation, however the underlying mechanisms are not fully understood. In this study, we explored the potential role of DNA-dependent protein kinase catalytic subunit (DNA-PKcs)-Akt signaling in LDI-induced osteoblast differentiation. We confirmed that LDI promoted mouse calvarial osteoblast differentiation, which was detected by increased alkaline phosphatase (ALP) activity as well as mRNA expression of type I collagen (Col I) and runt-related transcription factor 2 (Runx2). In mouse osteoblasts, LDI (1Gy) induced phosphorylation of DNA-PKcs and Akt (mainly at Ser-473). The kinase inhibitors against DNA-PKcs (NU-7026 and NU-7441) or Akt (LY294002, perifosine and MK-2206), as well as partial depletion of DNA-PKcs or Akt1 by targeted-shRNA, dramatically inhibited LDI-induced Akt activation and mouse osteoblast differentiation. Further, siRNA-knockdown of SIN1, a key component of mTOR complex 2 (mTORC2), also inhibited LDI-induced Akt Ser-473 phosphorylation as well as ALP activity increase and Col I/Runx2 expression in mouse osteoblasts. Co-immunoprecipitation (Co-IP) assay results demonstrated that LDI-induced DNA-PKcs-SIN1 complexation, which was inhibited by NU-7441 or SIN1 siRNA-knockdown in mouse osteoblasts. In summary, our data suggest that DNA-PKcs-SIN1 complexation-mediated Akt activation (Ser-473 phosphorylation) is required for mouse osteoblast differentiation.

Keloids are one of the common refractory conditions in dermatology and aesthetic plastic surgery. The most effective treatment is superficial radiotherapy followed by surgical removal. The rate of recurrence is strongly associated with the total dose of ionizing irradiation, and the underlying mechanism remains unclear. In this study, we used primary keloid fibroblasts (KFb) isolated from patient samples to investigate the effects of X-ray radiation on cell proliferation, cell toxicity and cell cycle, as detected by CCK-8 assay kit and flow cytometer. In addition, we examined senescence-associated β-galactosidase activity and the associated gene expression using real-time polymerase chain reaction and western blot in KFb exposed to X-ray radiation. X-ray radiation inhibited cell proliferation and induced cell senescence in KFb in a dose-dependent manner. Inhibition of cell proliferation and induction of cellular senescence were mediated by interruption of the cell cycle with an extended G0/G1 phase. Furthermore, the expressions of senescence-associated genes p21, p16 and p27 were upregulated both at mRNA and protein levels in KFb exposed to X-ray radiation. Taken together, our data indicate that X-ray radiation may prevent the recurrence of keloids by controlling fibroblast proliferation, arresting the cell cycle and inducing premature cellular senescence.

Femoral midshaft fracture is one of the most common clinical injuries and is often caused by high-energy traffic accidents. Intramedullary nailings, plates, and external fixators are all used as treatment alternatives for a variety of patients depending on fracture location, displacement, comminution, soft tissue condition, and local tradition. Locked intramedullary nailing is currently the preferred treatment method for most diaphyseal fractures and has good clinical results. The goal of this study was to compare expandable and locked intramedullary nailing for the treatment of AO type 32A and 32B1 femoral midshaft fractures. The authors performed a retrospective analysis of 46 patients (33 men and 13 women; mean age, 32.3 years; range, 22–52 years) with femoral midshaft fractures who were divided into 2 groups—one treated with an expandable intramedullary nailing method and the other with a conventional locked intramedullary nailing. The 2 groups were compared with respect to operation time, fluoroscopic time, amount of estimated blood loss, hospitalization time, healing time, and complications. Patients were followed for at least 1 year. The results of this study showed that all of the patients achieved bone union within 12 to 24 months. Expandable nailing performed better than locked nailing in operation time, fluoroscopic time, amount of estimated blood loss, and healing time ( P Orthopedics. 2015; 38(4):e314–e318.]

Prolonged or overdose glucocorticoids (GCs) usage is the common cause of osteoporosis. In the present study, we studied the cellular mechanism of dexamethasone (Dex)-induce osteoblast cell death by focusing on the role of mitochondrial permeability transition pore (mPTP). In cultured osteoblastic MC3T3-E1 cells, Dex-induced mPTP opening, which was demonstrated by mitochondrial membrane potential (MPP) decrease, cyclophilin-D (CyPD)-adenine nucleotide translocator 1 (ANT-1) mitochondrial complexation and cytochrome C (cyto-C) release. The mPTP inhibitor sanglifehrin A (SfA) dramatically inhibited Dex-induced MPP loss, cyto-C release and MC3T3-E1 cell death. Dex-induced cell death requires mPTP composing protein CyPD, as CyPD inhibitor cyclosporin A (CsA) and CyPD siRNA knockdown inhibited Dex-induced MC3T3-E1 cell death, while CyPD overexpression aggravated Dex's cytotoxic effect. We found that Dex induced P53 phosphorylation and translocation to mitochondria, where it formed a complex with CyPD. Glucocorticoid receptor (GR) siRNA knockdown, or P53 inhibition (by its inhibitor pifithrin-α or shRNA silencing) suppressed Dex-induced CyPD-P53 mitochondrial association and subsequent MC3T3-E1 cell death. Finally, in primary cultured osteoblasts, Dex-induced cell death was inhibited by CsA, SfA or pifithrin-α. Together, our data suggest that Dex-induced osteoblast cell death is associated with GR-P53-regulated mPTP opening.

Patients with age related macular degeneration (AMD) will develop vision loss in the center of the visual field. Reactive oxygen species (ROS)-mediated retinal pigment epithelium (RPE) cell apoptosis is an important contributor of AMD. In this study, we explored the pro-survival effect of α-melanocyte stimulating hormone (α-MSH) on oxidative stressed RPE cells. We found that α-MSH receptor melanocortin 1 receptor (MC1R) was functionally expressed in primary and transformed RPE cells. RPE cells were response to α-MSH stimulation. α-MSH activated Akt/mammalian target of rapamycin (mTOR) and Erk1/2 signalings in RPE cells, which were inhibited by MC1R siRNA knockdown. α-MSH protected RPE cells from hydrogen peroxide (H₂O₂)-induced apoptosis, an effect that was almost abolished when MC1R was depleted by siRNA. α-MSH-mediated S6K1 activation and pro-survival effect against H₂O₂ was inhibited by Akt inhibitors (perifosine, MK-2206 and LY294002). Further, mTOR inhibition by rapamycin, or by mTOR siRNA knockdown, diminished α-MSH's pro-survival effect in RPE cells. Thus, Akt and its downstream mTOR signaling mediates α-MSH-induced survival in RPE cells. In summary, we have identified a new α-MSH-MC1R physiologic pathway that reduces H₂O₂-induced RPE cell damage, and might minimize the risk of developing AMD.

Background and objectives Symptomatic ASD after lumbar spinal fusion surgery occurs most commonly in the cranial segment. The surgery for ASD contains anterior lumbar interbody fusion, posterior lumbar interbody fusion, decompression alone (laminotomy) and so on. But coblation nucleoplasty for ASD has not been reported previously. In this study, a case of coblation nucleoplasty after posterolateral fusion surgery at L4-L5 for adjacent segment degeneration (ASD) was reported and the clinical results were examined. Material and method A 32-year-old male patient who had discectomy and fusion on the L4-L5 level seven years ago complained of chronic back pain for four months with numbness on his right leg for a month. X-ray revealed mild lumbar instability on L3-L4 segment. Magnetic resonance imaging confirmed a right-sided L3-L4 herniated disc compressing the L4 nerve root. He underwent L3-L4 coblation nucleoplasty. The visual analog scale (VAS) was adopted to assess the relief of back pain, leg pain and numbness. Results The operation was performed successfully and the symptoms were relieved significantly at the follow-up of more than twenty-four months. Conclusion Although coblation nucleoplasty is not a regular therapy for ASD, the excellent outcome of this case suggests that this technique might be an option before a complicated revision surgery.

The purpose of this study was to evaluate longer-term efficacy over a two-year follow-up of coblation nucleoplasty treatment for protruded lumbar intervertebral disc.Forty-two cases of protruded lumbar intervertebral disc treated by coblation nucleoplasty followed-up for two years were analysed. Relief of low back pain, leg pain and numbness after the operation were assessed by visual analogue pain scale (VAS). Function of lower limb and daily living of patients were evaluated by the Oswestry Disability Index (ODI).Operations were performed successfully in all cases. Three patients had recurrence within a week of the procedure. Evaluation of the 42 patients demonstrated significant improvement rate of VAS: defined as 66.2% in back pain, 68.1% in leg pain, and 85.7% in numbness at one-week after the operation; 53.2%, 58.4%, 81.0% at one-year; and 45.5%, 50.7%, 75.0% at two-year follow-up. One week after the operation, obvious amelioration occurred in all the patients, but the tendency decreased. Before operation, the mean value of ODI was 68.2 ± 10.9%. The value at one week was 28.6 ± 8.2%; one-year at 35.8 ± 6.5%; and two-years at 39.4 ± 5.8%.Coblation nucleoplasty may have satisfactory clinical outcomes for treatment of protruded lumbar intervertebral disc for as long as two-year follow-up, but longer-term benefit still needs verification.

To evaluate the effect of local X-irradiation on spinal cord injury by using physiology, kinology, electrophysiology and histology method.46 female Sprague-Dawley rats were subjected to spinal cord injury by weight dropping on T(11-12). All animals were divided into 3 groups randomly. One of the animal groups was irradiated with 10 Gy at the lesion site; another was irradiated with 20 Gy, the other without irradiation is regarded as sham-group. The animals were euthanized at different time points at 4 and 12 weeks after irradiation. Spinal cord callus was assessed by using physiology, kinology, and electrophysiology and histology method.In all the groups, the NF at 14 weeks were found higher than that of 6 weeks. Both 10 Gy irradiated and 20 Gy irradiated groups were higher than those of group at each time point (P0.05). The MBP decreased at 14 weeks in irradiated groups (P0.05), but increased at 14 weeks in sham-group (P0.05), the MBP of irradiated groups was lower than that sham-group at 14 weeks (P0.05). The GFAP and Nogo-A at 14 weeks were higher than that in 6 weeks in all the groups (P0.05), and there was no statistical significance with physiology, kinology, electrophysiology test in all groups.A self-repair mechanism exists after SCI, which will last at least 14 weeks. Local irradiation promotes the regeneration of spinal cord system after injury to some extent.

To investigate the effect of low dose irradiation (LDI) on callus formation and mineralization.80 SD rats were subjected to standard closed fracture on right femur so as to establish animal models and then were randomly divided into 2 equal groups: LDI group undergoing low dose whole body X-ray irradiation of 1Gy right after fracture induction and control group without irradiation. 1, 2, 3, 4, and 8 weeks later blood samples were collected from the heart to examine the amounts of white blood cells and platelets and alkaline phosphatase (ALP) level. Conventional X-ray films were taken and MicroCT was conducted to evaluate the callus bridging. Then the rats were killed to take out the femurs to undergo histological examination. Peripheral quantitative computed tomography (pQCT) was conducted to quantify the bone mineral content (BMC). Four-point bending test was used to examine the mechanical properties of the callus.The maximum load value and histological score 2 weeks later of the LDI group were both lower than those of the control group, however, the serum ALP, BMC, maximum load level, callus bridging score, and histological score 3 weeks later of the LDI group were all significantly higher than those of the control groups (all P0.05). 4 and 8 weeks later there were not significant differences in all these parameters between the 2 groups. The WBC amount in the first 2 weeks of experiment of the LDI group was remarkably lower in these 2 groups, however, in the third week, the WBC value of the LDI group was higher than that of the control group (P0.05).LDI promotes the mineralization at the stage of hard callus formation which is probably associated with progenitor cell mobilization.

This study investigated the hypothesized beneficial effect of low-dose irradiation (LDI) on fracture callus mineralization in a rat model. Seventy-two male Sprague–Dawley rats were averagely randomized into LDI group (rats treated with LDI) and SHAM group (rats treated with sham irradiation). Right after either LDI or sham irradiation, a standardized closed fracture on the right femur was established. At 2, 3 and 4 weeks postfracture, 12 rats in each group were euthanized. Fracture callus was assessed by using radiography and MicroCT for callus bridging, peripheral quantitative computed tomography (pQCT) for quantifying bone mineral content (BMC) and cross sectional area (CSA), confocal laser scanning microscopy for measuring area fraction of fluorescence labeling (AFFL) and four-point bending test for examining mechanical properties. The CSA and AFFL were found to be 22 and 33% smaller in the LDI group compared to the SHAM group at 2 weeks (P

We compared the effectiveness of postural reduction and instrumental reduction in the treatment of thoracolumbar and lumbar vertebra fracture in 40 patients. Under general anaesthesia, postural reduction in a stretching prone position was first performed. Instrumental reduction and fixation were then conducted. Radiographs were made after each reduction. Comparisons between the two films and the fracture films were made based on the changes in prevertebral height of both the fractured vertebra and the adjacent superior and inferior intervertebral spaces. It was found that the recovery of the prevertebral height in postural and instrumental reductions was basically identical. The recovery of the prevertebral height in the intervertebral spaces was more significant in instrumental reduction. Both reductions were ineffective in patients whose compression of the diseased vertebra was more than two-thirds of the normal. In cases of lower lumbar vertebra fractures, the effect of both reductions was unsatisfactory. Our findings indicated that the effectiveness of the reduction of vertebra fracture depends on the quantitative change of the spongy bone of the injured vertebra. Instrumental reduction only exerts an indirect tension. Postural reduction is effective in reducing thoracolumbar vertebral fracture, while instrumental reduction exerts only a relatively weak effect but it is particularly useful to maintain the result of postural reduction.

To discuss the characteristics and operative selection of far lateral lumbar disc herniation (FLLDH).Twenty-three cases of FLLDH, 14 were foraminal, and 9 were extraforaminal lumbar disc herniation. Of the 23 cases, low back pain was observed in 8 cases (31%), severe lower leg pain in 21 cases (91%) and Lasegue sign in 10 cases (43%). CT and MRI showed the protruded disc in and outside of the foramen clearly. Three surgical procedures were performed, including hemilaminotomy with medial facetectomy, facetectomy with pedicle screw fixation and fusion with posteolateral bone grafting, and the transmuscular approaches.Twenty-two cases were followed up for an average of 3.6 years. According to the Macnab criteria, 15 patients achieved excellent results, good 4, fair 3 and poor 0. Excellent and good rate was 86%.The symptoms and signs of FLLDH mainly result from injury of upper nerve segments with the dominant symptom of severe lower leg pain. CT and MRI appearance are not only sensitive but also specific for the diagnosis of FLLDH. In foraminal lumbar disc herniation, the hemilaminotomy with medial facetectomy is recommended. While in extraforaminal lumbar disc herniation, either facetectomy with pedicle screw fixation and fusion with posterolateral bone grafting or transmuscular approaches for removal of nucleus pulposus can be chosen. Microendoscopic discectomy is a new, safe and efficient method for the disease, however, a skillful microendoscopic technique should be mastered prior.

We investigated the optical properties of LaB6 using first-principles calculations within the framework of density functional theory. It was found that our calculated results are in excellent agreement with the previously reported available and experimental findings. Analysis of the dielectric functions showed that LaB6 has the unique optical behavior in the visible and near infrared range and revealed that origins of solar heat-shielding performance and high visible transparency for LaB6 should be ascribed to its plasmonic behaviors.