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Wei Liang, Pengbing Ding, Guan Li, Enhang Lu, Zhenmin Zhao Department of Plastic Surgery, Peking University Third Hospital, Beijing, 100191, People’s Republic of ChinaCorrespondence: Zhenmin ZhaoDepartment of Plastic Surgery, Peking University Third Hospital, Beijing, 100191, People’s Republic of ChinaEmail zzmbysy@sina.comBackground: The potential of relapse of craniofacial disharmony after trans-sutural distraction osteogenesis is high due to the failure to produce a stable bone bridge in the suture gap. The aim of this study is to evaluate whether hydroxyapatite nanoparticles (nHAP) have the effect of promoting osteoblast differentiation of suture-derived stem cells (SuSCs) and bone formation in sagittal suture during expansion.Methods: SuSCs were isolated from sagittal sutures and exposed to various concentrations of nHAP (0, 25, 50, and 100 μg mL− 1) to determine the optimal concentration of nHAP in osteoblast differentiation via performing Western Blotting and RT-qPCR. Twenty 4-week-old male Sprague–Dawley rats were randomly assigned into 4 groups: SHAM (sham-surgery), distraction, ACS (absorbable collagen sponge) and ACS+nHAP groups. In the ACS and ACS+nHAP groups, saline solution and nHAP suspended in a saline solution were delivered by ACS placed across the sagittal suture, respectively. In the latter three groups, the suture was expanded for 14 days by 50 g of constant force via a W shape expansion device. Suture gap area, bone volume fraction (BV/TV) and bone mineral density (BMD) of sagittal sutures were assessed via micro-CT, while the mechanical properties of sagittal sutures were evaluated via nanoindentation test. The efficacy of nHAP on bone formation in sagittal suture was also evaluated via BMP-2 immunohistochemistry staining.Results: The expression of osteoblast related genes and proteins induced by 25μg mL− 1 nHAP were significantly higher than the other groups in vitro (p< 0.05). Furthermore, treating with 25μg mL− 1 nHAP in vivo, the suture gap area was significantly reduced when compared with the distraction group. Correspondingly, the BV/TV, BMD, hardness and modulus of sagittal sutures were significantly increased in the ACS+nHAP group (p< 0.05).Conclusion: The 25μg mL− 1 dose of nHAP delivered by ACS can facilitate bone formation into the sagittal suture during expansion via inducing osteoblast differentiation of SuSCs.Keywords: hydroxyapatite nanoparticles, suture-derived stem cells, osteoblast differentiation, sagittal suture, bone formation

Lulu Zhou,1 Miaomiao Ye,1 Fang Xue,1 Ermei Lu,1 Lu-Zhe Sun,2 Xueqiong Zhu11Department of Obstetrics and Gynecology, The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou 325027, People’s Republic of China; 2Departments of Cell Systems & Anatomy, School of Medicine, University of Texas Health Science Center, San Antonio, TX, USAObjective: To investigate dynein light chain Tctex-type 3 (DYNLT3) protein expression in ovarian epithelial lesions and explore the effects and related mechanisms of DYNLT3 in terms of the biological behavior of ovarian cancer.Materials and methods: Initially, expression of the DYNLT3 protein in ovarian epithelial lesions was detected by immunohistochemical staining, and the prognostic value of DYNLT3 mRNA expression in ovarian cancer patients was assessed using the Kaplan-Meier plotter database. Then, the mRNA and protein expression of DYNLT3 in IOSE80 normal ovarian epithelial cells and SKOV3 ovarian cancer cells was evaluated by quantitative real-time polymerase chain reaction and Western blotting respectively, and the proliferation, apoptosis, migration and invasion of SKOV3 cells after DYNLT3 over-expression and under-expression were investigated by CCK-8 assays and immunofluorescence staining, flow cytometry, wound healing assays and Transwell invasion assays, respectively. Furthermore, the expression of the proliferation-related proteins PCNA and Ki-67 and the invasion- and migration-related proteins Ezrin, Fascin, MMP2 and MMP9 in cells was examined by Western blotting.Results: The protein expression of DYNLT3 gradually increased during the progression of ovarian epithelial lesions, and was related to the development of ovarian cancer. High expression of DYNLT3 mRNA was related to poor overall survival and progression free survival, especially in serous ovarian cancer patients. In addition, overexpression of DYNLT3 promoted SKOV3 cell proliferation, invasion and migration. The corresponding results were also verified by a DYNLT3 knockdown assay. Moreover, DYNLT3 increased cell proliferation, which was related to Ki-67 expression. Besides, DYNLT3 enhanced cell invasion and migration through regulating Ezrin, but not Fascin, MMP2 or MMP9.Conclusion: DYNLT3 exerts pro-tumoral effects on ovarian cancer through promoting cell proliferation, migration and invasion, possibly via regulating the protein expression of Ki-67 and Ezrin. DYNLT3 may be a potential prognostic predictor in ovarian cancer.Keywords: DYNLT3, ovarian cancer, proliferation, invasion, migration, prognosis

Chuchu Zhao,* Ermei Lu,* Xiaoli Hu, Huihui Cheng, Jian-An Zhang, Xueqiong Zhu Department of Obstetrics and Gynecology, The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325027, China *These authors contributed equally to this work Objective: Our previous research has shown that the expression of S100 calcium-binding protein A9 (S100A9) in tumor cells was associated with neoadjuvant chemotherapy sensitivity in cervical squamous cell carcinoma. In the present study, we altered the expression of S100A9 through infecting lentivirus, investigated its effect on the chemosensitivity to cisplatin of cervical cancer cells and then made a primary exploration of the involved mechanism.Materials and methods: Lentivirus was employed to upregulate and downregulate S100A9 expression in SiHa cells. The protein expression level of apoptotic-related proteins Bcl-2 and Bax, drug resistance-related proteins multiple drug resistance protein 1 (MRP1), P glycoprotein (P-gp), glutathione-S-transferase-π (GST-π), lung resistance-related protein (LRP), and FOXO1 signaling pathway related proteins was detected by Western blot. The CCK-8 assay was used to examine chemosensitivity to cisplatin, and the proportion of apoptosis cells was analyzed by the flow cytometry.Results: S100A9 overexpression could obviously increase the IC50 value of SiHa cells to cisplatin and decrease the apoptosis rate induced by cisplatin. Downregulation of S100A9 led to the opposite results. In S100A9 overexpression SiHa cells, the expression level of Bcl-2, LRP, GST-π, p-AKT, p-ERK, p-FOXO1, and Nanog was significantly increased, while FOXO1 expression was decreased. The opposite results were observed in S100A9 knockdown SiHa cells.Conclusion: Downregulation of S100A9 could significantly increase apoptosis rate, resulting in enhancing sensitivity of SiHa cells to cisplatin, which may be related to Bcl-2, GST-π, and LRP protein and by altering the AKT/ERK-FOXO1-Nanog signaling pathway. Keywords: S100A9, cervical cancer, Bcl-2, GST-π, LRP, FOXO1, cisplatin, chemosensitivity

The prefrontal cortex (PFC) mediates higher cognition but is impaired by stress exposure when high levels of catecholamines activate calcium-cAMP-protein kinase A (PKA) signaling. The current study examined whether stress and increased cAMP-PKA signaling in rat medial PFC (mPFC) reduce pyramidal cell firing and impair working memory by activating KCNQ potassium channels. KCNQ2 channels were found in mPFC layers II/III and V pyramidal cells, and patch-clamp recordings demonstrated KCNQ currents that were increased by forskolin or by chronic stress exposure, and which were associated with reduced neuronal firing. Low dose of KCNQ blockers infused into rat mPFC improved cognitive performance and prevented acute pharmacological stress-induced deficits. Systemic administration of low doses of KCNQ blocker also improved performance in young and aged rats, but higher doses impaired performance and occasionally induced seizures. Taken together, these data demonstrate that KCNQ channels have powerful influences on mPFC neuronal firing and cognitive function, contributing to stress-induced PFC dysfunction. Keywords: cAMP-PKA, KCNQ, Pyramidal neurons, Working memory, Stress, Prefrontal cortex

Shuang Li,1,2 Weijun Yu,3 Weiqi Zhang,1 Guohua Zhang,1 Li Yu,1 Eryi Lu1 1Department of Stomatology, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, People’s Republic of China; 2Shanghai Key Laboratory of Orthopedic Implants, Department of Orthopedic Surgery, Shanghai Ninth People’s Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, People’s Republic of China; 3College of Stomatology, School of Medicine, Shanghai Jiao Tong University, Shanghai, People’s Republic of China Background: Optimal osseointegration has been recognized as a pivotal factor in determining the long-term success of biomedical implants. Materials and methods: In the current study, highly carbonated hydroxyapatite (CHA) with carbonate contents of 8, 12 and 16 wt% and pure hydroxyapatite (HA) were fabricated via a novel hydrothermal method and deposited on the titanium substrates to generate corresponding CHA bioceramic coatings (designated as C8, C12 and C16, respectively) and HA bioceramic coatings (designated as C0). Results: C8, C12 and C16 were endowed with nanoscale, hierarchical hybrid micro-/nanoscale and microscale surface topographies with rod-like superstructures, respectively. Compared with C0, the micro-/nanotextured CHA bioceramic coatings (C8, C12 and C16) possessed excellent surface bioactivity and biocompatibility, as well as better wettability, which mediated improved protein adsorption, giving rise to simultaneous enhancement of a biological cascade of events of rat bone-marrow-derived mesenchymal stem cells including cell adhesion, proliferation, osteogenic differentiation and, notably, the production of the pro-angiogenic growth factor, vascular endothelial growth factor-A. In particular, C12 with biomimetic hierarchical hybrid micro-/nanorod topography exhibited superior fractal property and predominant performance of protein adsorption, cell adhesion, proliferation and osteogenesis concomitant with angiogenesis. Conclusion: All these results suggest that the 12 wt% CHA bioceramic coating with synergistic modification of surface chemistry and topography has great prospect for future use as implant coating to achieve optimum osseointegration for orthopedic and dental applications. Keywords: carbonated hydroxyapatite, micro-/nanohybrid topography, biomimicry, titanium implant

Chenyuan Zhu,1,2,* Yuting Lv,3,4,* Chao Qian,1,2,* Zihao Ding,3,5 Ting Jiao,1,2 Xiaoyu Gu,1,2 Eryi Lu,6 Liqiang Wang,3 Fuqiang Zhang1,2 1Department of Prosthodontics, Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, 2Shanghai Key Laboratory of Stomatology, Shanghai Research Institute of Stomatology, National Clinical Research Center of Stomatology, 3State Key Laboratory of Metal Matrix Composites, Shanghai Jiao Tong University, Shanghai, 4College of Mechanical and Electronic Engineering, Shandong University of Science and Technology, Qingdao, People’s Republic of China; 5Department of Materials Science and Engineering, Carnegie Mellon University, Pittsburgh, PA, USA; 6Department of Stomatology, Ren Ji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, People’s Republic of China *These authors contributed equally to this work Background: The interaction between the material and the organism affects the survival rate of the orthopedic or dental implant in vivo. Friction stir processing (FSP) is considered a new solid-state processing technology for surface modification. Purpose: This study aims to strengthen the surface mechanical properties and promote the osteogenic capacity of the biomaterial by constructing a Ti-6Al-4V (TC4)/zinc (Zn) surface nanocomposites through FSP. Methods: FSP was used to modify the surface of TC4. The microstructures and mechanical properties were analyzed by scanning electron microscopy, transmission electron microscopy, nanoindentation and Vickers hardness. The biological properties of the modified surface were evaluated by the in vitro and in vivo study. Results: The results showed that nanocrystalline and numerous β regions, grain boundary a phase, coarser acicular α phase and finer acicular martensite α' appeared because of the severe plastic deformation caused by FSP, resulting in a decreased elastic modulus and an increased surface hardness. With the addition of Zn particles and the enhancement of hydrophilicity, the biocompatibility was greatly improved in terms of cell adhesion and proliferation. The in vitro osteogenic differentiation of rat bone marrow stromal cells and rapid in vivo osseointegration were enhanced on the novel TC4/Zn metal matrix nanocomposite surface. Conclusion: These findings suggest that this novel TC4/Zn surface nanocomposite achieved by FSP has significantly improved mechanical properties and biocompatibility, in addition to promoting osseointegration and thus has potential for dental and orthopedic applications. Keywords: nanocomposite, friction stir processing, nanocrystalline/ultrafine grained, bone marrow stromal cells, cell proliferation, osteogenic differentiation

Yaqin Wu,1–3,* Anpeng Wang,1,3,4,* Biqing Zhu,1–3 Jian Huang,2,3 Emei Lu,2,3 Hanzi Xu,2,3 Wenjie Xia,1,3,4 Gaochao Dong,1 Feng Jiang,1,3,4 Lin Xu1,3,4 1Jiangsu Key Laboratory of Molecular and Translational Cancer Research, Jiangsu Cancer Hospital, Jiangsu Institute of Cancer Research, The Affiliated Cancer Hospital of Nanjing Medical University, Nanjing, People’s Republic of China; 2Department of Radiation Oncology, Jiangsu Cancer Hospital, Jiangsu Institute of Cancer Research, The Affiliated Cancer Hospital of Nanjing Medical University, Nanjing, People’s Republic of China; 3The Fourth Clinical College of Nanjing Medical University, Nanjing, People’s Republic of China; 4Department of Thoracic Surgery, Jiangsu Cancer Hospital, Jiangsu Institute of Cancer Research, The Affiliated Cancer Hospital of Nanjing Medical University, Nanjing, People’s Republic of China*These authors contributed equally to this work Background: KIF18B was identified as a potential oncogene by analysis of The Cancer Genome Atlas database.Materials and methods: We assessed KIF18B expression and explored its clinical significance in cervical cancer tissues. We have also evaluated the effects of KIF18B on cervical cancer cell proliferation, migration, and invasion both in vitro and in vivo.Results: Our results show that KIF18B is overexpressed in cervical cancer tissues and is associated with a large primary tumor size, an advanced FIGO stage, and an advanced tumor grade. Knockdown of KIF18B induces cell cycle G1-phase arrest and inhibits the proliferation, migration, and invasion of cervical cancer cells, whereas its overexpression promotes proliferation, migration, and invasion in these cells. Moreover, silencing of KIF18B reduces expression of CyclinD1, β-catenin, C-myc, and p-GSK3β expression.Conclusion: These data suggest that KIF18B can serve as a novel oncogene that promotes the tumorigenicity of cervical cancer cells by activating Wnt/β-catenin signaling pathway. Keywords: KIF18B, cervical cancer, CyclinD1, Wnt/β-catenin signaling pathway

Radiotherapy using protons and heavier ions is emerging as an alternative to traditional photon radiotherapy for cancer treatment. Ions have a depth-dose profile that results in high energy deposition at the end of the particle’s path, with a relatively low dosage elsewhere. However, the specifics of ion interactions with cellular biology are not yet fully understood. To study the induced biological effects of the ions on cell cultures, an external beam is required as biological specimens cannot be placed in vacuum. The Heavy Ion Accelerator Facility (HIAF) at the Australian National University hosts accelerators for a wide variety of ion-beam research applications. However, HIAF does not currently have an external beam capability. Here, we present an initial design for a radiobiological research capability at HIAF. A systems engineering approach was used to develop the architecture of the apparatus and determine the feasibility of adapting the current facilities to external beam applications. This effort included ion optics calculations, coupled to a Geant4 simulation, to characterise ion beam transitions through a thin window into the air. The beam spread, intensity distributions, and energy of proton and carbon ions were studied as a function of distance travelled from the window, as well as the effects of alternative window materials and thicknesses. It was determined that the proposed line at the HIAF would be suitable for the desired applications. Overall, this feasibility study lays the foundations of an external beam design, a simulation test framework, and the basis for a grant application for an external beam at the HIAF.

Jinjun Ye,1,* Li Yin,1,* Peng Xie,1 Jianfeng Wu,1 Jian Huang,1 Guoren Zhou,2 Hanzi Xu,1 Emei Lu,1 Xia He1 1Department of Radiotherapy, 2Department of Chemotherapy, Affiliated Jiangsu Cancer Hospital of Nanjing Medical University, Nanjing, People’s Republic of China *These authors contributed equally to this work Abstract: We investigated the effects of troglitazone on human cervical cancer SiHa cells and its mechanisms of action. SiHa cells were incubated with different concentrations of troglitazone (100, 200, or 400 μg/mL) for 24, 48, and 72 hours. Cell viability was measured by 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT) assay; cell cycle and apoptosis were detected by flow cytometry; and morphology of SiHa cells was observed under an inverted microscope. pcDNA3.1 and pcDNA3.1-Skp2 plasmids were constructed and then transfected into SiHa cells. Protein expression was analyzed by Western blotting. Troglitazone inhibited the proliferation of SiHa cells in a time- and concentration-dependent manner. Troglitazone caused G0/1 phase arrest but failed to reduce apoptosis in SiHa cells. Troglitazone significantly increased expression of p27 but decreased Skp2 expression. Skp2 overexpression inhibited the role of troglitazone in increasing expression of p27, and the cell cycle inhibitory effect of troglitazone. Troglitazone can inhibit SiHa cell viability by affecting cell cycle distribution but not apoptosis, and Skp2 and p27 may play a critical role. Keywords: SiHa cells, cell cycle, apoptosis

Yanzhi Song,1 Zhenjun Huang,1 Yang Song,2 Qingjing Tian,1 Xinrong Liu,1 Zhennan She,1 Jiao Jiao,1 Eliza Lu,3 Yihui Deng11College of Pharmacy, Shenyang Pharmaceutical University, Shenyang, People’s Republic of China; 2Jiangsu Hansoh Pharmaceutical Co., Ltd., Lianyungang, People’s Republic of China; 3Livzon Mabpharm Inc., Zhuhai, People’s Republic of ChinaAbstract: The applications of ethylenediaminetetraacetic acid (EDTA) have been expanded from the treatment of heavy metal poisoning to chelation therapies for atherosclerosis, heart disease, and cancers, in which EDTA reduces morbidity and mortality by chelating toxic metal ions. In this study, EDTA was used in a drug delivery system by adopting an NH4EDTA gradient method to load doxorubicin into liposomes with the goal of increasing therapeutic effects and decreasing drug-related cytotoxicity. The particle size of the optimum NH4EDTA gradient liposomes was 79.4±1.87 nm, and the entrapment efficiency was 95.54%±0.59%. In vitro studies revealed that liposomes prepared using an NH4EDTA gradient possessed long-term stability and delayed drug release. The in vivo studies also showed the superiority of the new doxorubicin formulation. Compared with an equivalent drug dose (5 mg/kg) prepared by (NH4)2SO4 gradient, NH4EDTA gradient liposomes showed no significant differences in tumor inhibition ratio, but cardiotoxicity and liposome-related immune organ damage were lower, and no drug-related deaths were observed. These results show that use of the NH4EDTA gradient method to load doxorubicin into liposomes could significantly reduce drug toxicity without influencing antitumor activity.Keywords: NH4EDTA, liposome, doxorubicin, ion gradient, antitumor activity, toxicity

The medial prefrontal cortex (mPFC) plays a key role in behavioral variability, action monitoring, and inhibitory control. The functional role of mPFC may change over the lifespan due to a number of aging-related issues, including dendritic regression, increased cAMP signaling, and reductions in the efficacy of neuromodulators to influence mPFC processing. A key neurotransmitter in mPFC is norepinephrine. Previous studies have reported aging-related changes in the sensitivity of mPFC-dependent tasks to noradrenergic agonist drugs, such as guanfacine. Here, we assessed the effects of yohimbine, an alpha-2 noradrenergic antagonist, in cohorts of younger and older rats in a classic test of spatial working memory (using a T maze). Older rats (23-29 mo.) were impaired by a lower dose of yohimbine compared to younger animals (5-10 mo.). To determine if the drug acts on alpha-2 noradrenergic receptors in mPFC and if its effects are specific to memory-guided performance, we made infusions of yohimbine into mPFC of a cohort of young rats (6 mo.) using an operant delayed response task. The task involved testing rats in blocks of trials with memory- and stimulus-guided performance. Yohimbine selectively impaired memory-guided performance. Infusions of muscimol (a GABA-A agonist) at the same sites selectively impaired memory-guided performance, but did not lead to error perseveration. Based on these results, we propose several potential interpretations for the role for the noradrenergic system in the performance of delayed response tasks, including the encoding of previous response locations, task rules (i.e. using a win-stay strategy instead of a win-shift strategy), and performance monitoring (e.g. prospective encoding of outcomes).

Background and Objectives: Vitamin D binding protein (DBP) is biosynthesised in the liver and is predominantly expressed in serum. Its primary role centres on facilitating the systemic transportation of vitamin D and its metabolites, notably 25‐hydroxyvitamin D, to specific target tissues where vitamin D exerts its biological functions. Due to the paucity of studies, it is unclear whether there is an association between DBP and periodontal status and thus its potential use as a diagnostic biomarker. Therefore, the aim of the systematic review is to investigate the association between DBP in periodontal disease. Methods: Two independent reviewers (YD and RG) performed a systematic literature search of English publications using several databases including MEDLINE (OVID interface, 1946 onwards), EMBASE (OVID interface, 1974 onwards), and Global Health (OVID interface, 1973 onwards). This search strategy enabled the identification of relevant publications and the development of a comprehensive library of studies. Studies were included based on previously agreed eligibility criteria. Of the eight studies included as part of this systematic review, seven were case–control studies and one was a cross‐sectional study. The quality assessment was based on the Newcastle–Ottawa Scale (NOS) for case–control studies and the modified NOS for the cross‐sectional study. Results: The NOS quality assessment was 'favorable' for 6 included case control studies; and 'fair' for one study. The modified NOS quality assessment for the single cross‐sectional study demonstrated a medium risk of bias. The results of the majority of the included studies indicated a statistically significant higher concentration of DBP levels in individuals with periodontitis in comparison to those who were periodontally healthy. This trend held true irrespective of the sampling method employed for the assessment of DBP concentration. Conclusion: The results summarised in this systematic review indicate a positive association between DBP and periodontitis. Nonetheless, there is a need for longitudinal, prospective trials, to confirm the use of DBP as a potential biomarker for the diagnosis of periodontitis. [ABSTRACT FROM AUTHOR]

The viability and high survival rate of probiotics through human gastrointestinal tract are very important to the host health. The present research was investigated to explicate the influence of encapsulation and wall materials on the survival of Enterococcus faecalis HZNU S1. Purposely, the microbeads were prepared using sodium alginate (SA) and flaxseed milk (FM) by extrusion method. The viability of investigated probiotics was accessed in simulated gastric juice (SGJ). Free cells exhibited a poor survival rate in SGJ. However, encapsulation improved the probiotic viability in the same circumstance. Encapsulated probiotics exhibited higher viability when they were exposed to pH 2.5 SGJ and 1.0 and 2.0% bile salt solutions. Furthermore, encapsulation could improve the storage stability of probiotics. The viable number of encapsulated cells was 10.01 Log CFU/g at 0 day and decreased to 9.6 Log CFU/g (4 °C) and 9.2 Log CFU/g (25 °C) after 8 days. Full encapsulated cells could be liberated after the microbeads were exposed to simulated intestine juice (SIJ) within 120 min. This study indicates that FM-SA microbeads can be suggested as good agents for encapsulating probiotics. [ABSTRACT FROM AUTHOR]

This study used a simple one-step hydrothermal method to synthesize the MoS2/GR composites with a new morphology composed of graphene nanotubes and ultra-thin molybdenum disulfide with the help of sodium chloride. The composites were characterized by XRD, XPS, SEM, TEM, and a series of characterization methods. Meanwhile, the tribological properties of the composites were studied. The results show that the addition of 1% MoS2/GR composite nanotubes has excellent tribological properties. In addition, the structure and excellent tribological properties of MoS2-C-nanocomposite lubrication materials will be conducive to designing new nanomaterials with 2D/3D structures, enhancing the anti-friction and anti-wear properties, and expanding their practical applications in industrial and agricultural fields. [ABSTRACT FROM AUTHOR]