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We investigated the anticancer effect of the aptamer-conjugated gemcitabine-loaded atelocollagen patch in a pancreatic cancer patient-derived xenograft (PDX) model to propose a future potential adjuvant surgical strategy during curative pancreatic resection for pancreatic cancer. A pancreatic cancer PDX model was established. Animals were grouped randomly into a no-treatment control group; treatment group treated with intraperitoneal gemcitabine injection (IP-GEM) or aptamer-conjugated gemcitabine (APT:GEM); and transplant with three kinds of patches: atelocollagen-aptamer-gemcitabine (patch I), atelocollagen-inactive aptamer-gemcitabine (patch II), and atelocollagen-gemcitabine (patch III). Tumor volumes and response were evaluated based on histological analysis by HE staining and Immunohistochemistry (IHC) was performed. Anticancer therapy-related toxicity was evaluated by hematologic findings. The patch I group showed the most significant reduction of tumor growth rate, compared with the no-treatment group (p 0.05). However, other treatment groups were not found to show significant reduction in tumor growth rate (0.05 p 0.1). There was no microscopic evidence suggesting potential toxicity, such as inflammation, nor necrotic changes in liver, lung, kidney, and spleen tissue. In addition, no leukopenia, anemia, or neutropenia was observed in the patch I group. This implantable aptamer-drug conjugate system is thought to be a new surgical strategy to augment the oncologic significance of margin-negative resection in treating pancreatic cancer in near future.

Benign paroxysmal positional vertigo (BPPV), the most common vestibular disorder, is diagnosed by an examiner changing the posture of the examinee and inducing nystagmus. Among the diagnostic methods used to observe nystagmus, video-nystagmography has been widely used recently because it is non-invasive. A specialist with professional knowledge and training in vertigo diagnosis is needed to diagnose BPPV accurately, but the ratio of vertigo patients to specialists is too high, thus necessitating the need for automated diagnosis of BPPV. In this paper, a convolutional neural network-based nystagmus extraction system, ANyEye, optimized for video-nystagmography data is proposed in the first study on automating BPPV diagnosis. A pupil was segmented to track the exact pupil trajectory from real-world data obtained during field inspection. A deep convolutional neural network model was trained with the new video-nystagmography dataset for the pupil segmentation task, and a compensation algorithm was designed to correct pupil position. In addition, a slippage detection algorithm based on moving averages was designed to eliminate the motion artifacts induced by goggle slippage. ANyEye outperformed other eye-tracking methods including learning and non-learning based algorithms with five-pixel error detection rate of 91.26%.

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Peroxiredoxin-3 (Prdx3) is a mitochondrial protein of the thioredoxin family of antioxidant peroxidases and is the principal peroxidase responsible for metabolizing mitochondrial hydrogen peroxide. Recent reports have shown that mitochondrial reactive oxygen species (mROS) contribute to macrophage-mediated bactericidal activity in response to Toll-like receptors. Herein, we investigated the functional effect of Prdx3 in bactericidal activity. The mitochondrial localization of Prdx3 in HEK293T cells was confirmed by cell fractionation and confocal microscopy analyses. To investigate the functional role of Prdx3 in bactericidal activity, Prdx3-knockdown (Prdx3KD) THP-1 cells were generated. The mROS levels in Prdx3KD THP-1 cells were significantly higher than those in control THP-1 cells. Moreover, the mROS levels were markedly increased in response to lipopolysaccharide. Notably, the Salmonella enterica serovar Typhimurium infection assay revealed that the Prdx3KD THP-1 cells were significantly resistant to S. Typhimurium infection, as compared with control THP-1 cells. Taken together, these results indicate that Prdx3 is functionally important in bactericidal activity through the regulation of mROS.

Given the high prevalence of atherothrombotic disease and increasing attention to functional foods, we tested the hypothesis that a combination supplement (PS) containing Phellinus baumii (PB) and Salvia miltiorrhiza Bunge (SM) may improve the atherothrombotic profile in rats. The PS (Experiment 1) or PB/SM (Experiment 2) supplement was administered orally once daily for 7 days prior to inducing platelet activation at the last day. Orally administered PS showed an additive and positive effect on nitric oxide (NO) availability, adhesion molecule/pro-inflammatory cytokine release, and thrombosis compared to controls. This effect was attributed more to PB than to SM for cyclooxygenase (COX)-1 and to SM for COX-2. In vitro data revealed that tanshinone IIA derived from SM remarkably suppressed COX-2, and protocatechuic acid derived from PB caused a small increase in NO production. The data suggest the potential use of PS to ameliorate endothelial and vascular dysfunction induced by unintended platelet activation.

Background and Objective The dielectrophoresis (DEP) technique is increasingly being recognised as a potentially valuable tool for non-contact manipulation of numerous cells as well as for biological single cell analysis with non-invasive characterisation of a cell's electrical properties. Several studies have attempted to track multiple cells to characterise their cellular DEP mobility. However, they encountered difficulties in simultaneously tracking the movement of a large number of individual cells in a bright-field image sequence because of interference from the background electrode pattern. Consequently, this present study aims to develop an automatic system for imaging-based characterisation of cellular DEP mobility, which enables the simultaneous tracking of several hundred of cells inside a microfluidic device. Methods : The proposed method for segmentation and tracking of cells consists of two main stages: pre-processing and particle centre localisation. In the pre-processing stage, background subtraction and contrast enhancement were performed to distinguish the cell region from the background image. In the particle centre localisation stage, the unmarked cell was automatically detected via graph-cut algorithm-based K-means clustering. Results : Our algorithm enabled segmentation and tracking of numerous Michigan Cancer Foundation-7 (MCF-7) cell trajectories while the DEP force was oscillated between positive and negative. The cell tracking accuracy and cell count capability was at least 90% of the total number of cells with the newly developed algorithm. In addition, the cross-over frequency was measured by analysing the segmented and tracked trajectory data of the cellular movements caused by the positive and negative DEP force. The measured cross-over frequency was compared with previous results. The multi-cellular movements investigation based on the measured cross-over frequency was repeated until the viability of cells was unchanged in the same environment as in a microfluidic device. The results were statistically consistent, indicating that the developed algorithm was reliable for the investigation of DEP cellular mobility. Conclusion : This study developed a powerful platform to simultaneously measure the DEP-induced trajectories of numerous cells, and to investigate in a robust, efficient, and accurate manner the DEP properties at both the single cell and cell ensemble level.

TRAF6 (TNF receptor associated factor 6) plays a pivotal role in NFKB activation and macroautphagy/autophagy activation induced by TLR4 (toll like receptor 4) signaling. The objective of this study was to determine the functional role of PRDX1 (peroxiredoxin 1) in NFKB activation and autophagy activation. PRDX1 interacted with the ring finger domain of TRAF6 and inhibited its ubiquitin-ligase activity. The inhibition on TRAF6 ubiquitin-ligase activity by PRDX1 induced the suppression of ubiquitination of an evolutionarily conserved signaling intermediate in Toll pathways (ECSIT) essential for NFKB activation and BECN1 (beclin 1) required for autophagy activation. An inhibitory effect of PRDX1 on TRAF6 was clearly evidenced in PRDX1-knockdown (PRDX1KD) THP-1, PRDX1KD MDA-MB-231, and PRDX1KD SK-HEP-1 cells. PRDX1KD THP-1 cells showed increases of NFKB activation, pro-inflammatory cytokine production, NFKB-dependent gene expression induced by TLR4 stimulation, and resistance against Salmonella typhimurium infection. Additionally, migration and invasion abilities of PRDX1KD MDA-MB-231 and PRDX1KD SK-HEP-1 cancer cells were significantly enhanced compared to those of control cancer cells. Taken together, these results suggest that PRDX1 negatively regulates TLR4 signaling for NFKB activation and autophagy functions such as bactericidal activity, cancer cell migration, and cancer cell invasion by inhibiting TRAF6 ubiquitin-ligase activity. Abbreviations: 3-MA: 3-methyladenine; BECN1: beclin 1; CHUK/IKKA: conserved helix-loop-helix ubiquitous kinase; ECSIT: ECSIT signalling integrator; ELISA: enzyme-linked immunosorbent assay; NFKB: nuclear factor kappa-light-chain-enhancer of activated B cells; IB: immunoblotting; IKBKB/IKKB: inhibitor of nuclear factor kappa B kinase subunit beta; IL1B: interleukin 1 beta; IL6: interleukin 6; IP: immunoprecipitation; LPS: lipopolysaccharide; MAP1LC3/LC3: microtuble associated protein 1 light chain 3; MAP3K7/TAK1: mitogen-activated protein kinase kinase kinase 7; MAPK14/p38: mitogen-activated protein kinase 14; mROS: mitochondrial reactive oxygen species; PRDX1: peroxiredoxin 1; PRDX6: peroxiredoxin 6; RELA/p65: RELA proto-oncogene, NF-kB subunit; TRAF6 TNF: receptor associated factor 6.

Injury to the peripheral axons of sensory neurons strongly enhances the regeneration of their central axons in the spinal cord. It remains unclear on what molecules that initiate such conditioning effect. Because ATP is released extracellularly by nerve and other tissue injury, we hypothesize that injection of ATP into a peripheral nerve might mimic the stimulatory effect of nerve injury on the regenerative state of the primary sensory neurons. We found that a single injection of 6 μl of 150 μm ATP into female rat sciatic nerve quadrupled the number of axons growing into a lesion epicenter in spinal cord after a concomitant dorsal column transection. A second boost ATP injection 1 week after the first one markedly reinforced the stimulatory effect of a single injection. Single ATP injection increased expression of phospho-STAT3 and GAP43, two markers of regenerative activity, in sensory neurons. Double ATP injections sustained the activation of phospho-STAT3 and GAP43, which may account for the marked axonal growth across the lesion epicenter. Similar studies performed on P2X7 or P2Y2 receptor knock-out mice indicate P2Y2 receptors are involved in the activation of STAT3 after ATP injection or conditioning lesion, whereas P2X7 receptors are not. Injection of ATP at 150 μm caused little Wallerian degeneration and behavioral tests showed no significant long-term adverse effects on sciatic nerve functions. The results in this study reveal possible mechanisms underlying the stimulation of regenerative programs and suggest a practical strategy for stimulating axonal regeneration following spinal cord injury. SIGNIFICANCE STATEMENT Injury of peripheral axons of sensory neurons has been known to strongly enhance the regeneration of their central axons in the spinal cord. In this study, we found that injection of ATP into a peripheral nerve can mimic the effect of peripheral nerve injury and significantly increase the number of sensory axons growing across lesion epicenter in the spinal cord. ATP injection increased expression of several markers for regenerative activity in sensory neurons, including phospho-STAT3 and GAP43. ATP injection did not cause significant long-term adverse effects on the functions of the injected nerve. These results may lead to clinically applicable strategies for enhancing neuronal responses that support regeneration of injured axons.

The anti-inflammatory effects of Acanthopanax senticosus leaves (ASL) and the effects of extrusion were evaluated. ASL exhibited in vitro and in vivo antiinflammatory activities in experimental systems. Extrusion increased the effect of ASL. ASL reduced C48/80-induced histamine release from HMC-1 cells as well as in vivo model, suggesting that ASL induces mast cell stabilization and has an anti-histamine activity. The effects of ASL and extruded ASL (ASLE) on pro-inflammatory cytokine production were evaluated. ASL treatment reduced MCP- 1, TNF-α, and IL-1β mRNA expressions and decreased their protein levels in HMC-1 cells. Decreases in serum NO, MDA, and TNF-α levels were observed in acute inflammatory rats, and extrusion increased the effects of ASL in a dose-dependent manner. These data support pharmacological basis of ASL and the effect of extrusion for future treatment of inflammation.

Aims The purposes of this study were to determine whether Cervi Pantotrichum Cornu (CPC) has osteogenic activities in human osteoblastic MG-63 cells and to investigate the underlying molecular mechanism. Main methods The effects of CPC on alkaline phosphatase activity, collagen synthesis, and calcium deposits were measured. The COL1A1 , ALPL , BGLAP , and SPP1 expressions were measured by real-time PCR. Phosphorylated MAP kinases (ERK1/2, JNK1/2, p38, ELK1, and cJUN) were studied by western blot analysis. The involvement of MAPK pathway in osteogenic gene expressions was determined by using each selective MAPK inhibitor (PD98059, SP600125, and SB203580). Key findings CPC increased alkaline phosphatase activity, collagen synthesis, and calcium deposits. CPC activated ERK1/2, JNK1/2, p38, and ELK1 phosphorylation except cJUN. CPC increased the COL1A1 , ALPL , BGLAP , and SPP1 gene expressions. The elevated COL1A1 and BGLAP expressions were inhibited by PD98059, SP600125 or SB203580. The elevated ALPL expression was blocked by SB203580. The elevated SPP1 expression was inhibited by SP600125 or SB203580. CPC increased COL1A1 and BGLAP expressions via ERK1/2, JNK1/2, and p38 MAPKs pathways and SPP1 expression via JNK1/2 and p38 pathways. p38 pathway is needed for ALPL expression. Significance These results imply that MAPK signaling pathway is an indispensable factor for bone matrix genes expression of CPC in MG-63 human osteoblast-like cells.

Collagen hydrolysates (CHs) are mixtures of peptides obtained by partial hydrolysis of gelatin that are receiving scientific attention as potential oral supplements for the restoration of osteoarticular tissues. The aim of this study was to evaluate the effectiveness of CHs for promoting longitudinal bone growth in growing rats. An in vitro study was carried out in osteoblast-like MG63 cells and the most effective CH on bone formation was selected among 36 various CHs. An in vivo study confirmed the functional effects of a selected CH with molecular weight of3 kDa on longitudinal bone growth. CHs dose-dependently promoted the longitudinal bone growth and height of the growth plate in adolescent male rats, whereas gelatin failed to affect longitudinal bone growth. Insulin-like growth factor-1 and bone morphogenetic protein-2 in the CH treated group were highly expressed in the growth plate. These results suggest that CHs isolated in this study may provide beneficial effects on bone metabolism of growing animals and humans.

Objectives： Bletillae Rhizoma, the roots of Bletilla striata, is used to restrain the leakage of blood and stop bleeding. It can cure the sores, ulcers, and chapped skin. This study was designed to investigate the collagen metabolism, elastase and tyrosinase activity of Bletillae Rhizo ma extract (BR).Methods : The effects of BR on type I procollagen production and collagenase activity in human normal fibroblasts Hs68 after UVB (312 nm) irradiation were measured b y ELISA method. The elastase activity, tyrosinase activity, and L‐DOPA oxidation after treatment of BR were measured as well. Results : In the present study, the collagen production (type I procollagen) was significantly increased to 15.7 ± 1.8 ng/ml at a concentration of BR 100 µg/ml in UVB damaged Hs68 cells. The increased collagenase activity after UVB damage was significantly recovered to 42.7 ± 0.7%, 54.5 ± 3.5%, and 38.4 ± 0.9% by BR 10, 30, and 100 µg/ml. The activities of BR 10 mg/ml on tyrosinase activity was significantly reduced to 45.1 ± 8.4% as well. However, there were no significant effects on the elastase activity and the L‐DOPA oxidation.Conclusion : BR showed the promoting effects of collagen synthesis and inhi bitory effects of collagenase activity in Hs68, human normal fibroblast cells. And these could be thought to have the anti‐wrinkle effects and whitening effects in vitro. These results suggest that BR may have potential as an anti ‐aging ingredient in cosmetic treatment. Key words : Bletillae Rhizoma, Elastase, Collagenase, Tyrosinase, Collagen , Hs68, Fibroblasts

Objectives : Coptidis Rhizoma has been used for stomach disease. However, i ts property is so cold that it might be avoided to prescribe for the elderly and the infirm having i ndigestion or diarrhea. Accordingly, the present study was designed to investigate the protective effects of Coptidis Rhizoma herbal acupuncture extract against acute gastric mucosal lesions induced by compound 48/80 in rats .Methods : The Coptidis Rhizoma herbal acupuncture (CRHA) was injected in Choksamni and Chungwan 1 h before compound 48/80 treatment. The animals were sacrificed under anesthesia 3 h after compound 48/80 treatment. The stomachs were removed and the amount of gastric adherent mucus, gastric mucosal hexosamine, SOD, XO, TBARS and histological examination were performed. Results : The decline of gastric adherent mucus, gastric mucosal hexosamine and the histological defects of gastric mucus were significantly protected by CRHA treatment. Gastric adherent mucus in control group was reduced to 38.2 ± 5.0%. CRHA groups significantly protected the loss of mucus to 77.5 ± 4.9%. Mucosal hexosamine content showed similar patterns. Mucosal hexosamine content in control group was reduced to 45.2 ± 6.2%. CRHA groups significantly protected the loss of mucus t o 83.0 ± 7.0%. The changes of gastric mucosal SOD and TBARS were recovered by CRHA treatment as well. Conclusions : CRHA showed the protective effects on the acute gastric mucosal lesions induced by compound 48/80 in rats. These results suggest that CRHA may have protective effects on the gastritis.Key words : Coptidis Rhizoma herbal acupuncture, Choksamni (ST36), Chungwa n (CV12), Acute gastric mucosal lesion, Compound 48/80

The berry of Panax ginseng significantly inhibited the histamine releases at the concentration of 30 μg/mL (p

【Objectives : This study was designed to investigate the collagen metabolism and tyrosinase activity of Sipgeondaebo-tang Pharmacopuncture extracts (SP). Methods : The effect of SP on type I procollagen production and collagenase activity in human normal fibroblasts HS68 after UVB (312 nm) irradiation was measured by ELISA method. The tyrosinase activity after treatment of SP was measured as well. Results : Type I procollagen production was recovered by SP in UVB damaged HS68 cells. The increased collagenase activity after UVB damage was significantly recovered by SP. The tyrosinase activity was significantly reduced as well. However, the L-DOPA oxidation was not changed. Conclusion : SP showed the anti-wrinkle effects and whitening effects in vitro. These results suggest that SP may be a potential pharmacopuncture as an anti-aging pharmacopuncture treatments.】

Objectives : Elastic fibers are found in the skin, lungs, arteries, veins and other structures. Elastases destroy the elastic fibers and cause the emphysema and pulmonary hypertension. Oxidative stress is needed for these pathologic changes. Accordingly, present study was designed to investigate the effect of Yukmigeehwang-hwan extracts (YHE) on elastase activity and anti-oxidative effects of YHE. Methods : The inhibitory effects on elastase and DPPH and NO free radical scavenging activities of YHE were measured. Results : The elastase activity was significantly inhibited by YHE. YHE significantly scavenged DPPH and NO free radicals as well. Conclusion : YHE showed the elastase-inhibiting effects and anti-oxidative activities in vitro. These results suggest that YHE may have potential roles in the treatment of pulmonary emphysema and pul- monary hypertension.

This study examined the interfacial adhesion, mechanical, and thermal properties of compatibilizing agent-treated and non-treated biocomposites as a function of the type of compatibilizing agent. The tensile strength, interfacial adhesion, and heat deflection temperature (HDT) of maleic anhydride-grafted poly(butylene succinate) (PBS-MA) and maleic anhydride-grafted poly(lactic acid) (PLA-MA)-treated biocomposites are greater than those of untreated maleic anhydride-grafted poly(styrene-b-ethylene-co-butylene-b-styrene) triblock copolymer (SEBS-MA) and maleic anhydride-grafted polypropylene (MAPP)-treated biocomposites. The storage modulus (E′) values and the tan δmax temperatures (Tg) of PBS-MA and PLA-MA-treated biocomposites were slightly higher than that of the untreated biocomposites.

It is well established that the IFN-gamma/STAT1 pathway plays an important role in the pancreatic beta-cell apoptosis that is observed in STZ-induced type 1 diabetes; however, the upstream regulatory proteins involved have not been understood. Here, we investigated whether activating transcription factor 3 (ATF3) affects STAT1-mediated beta-cell dysfunction and apoptosis in streptozotocin-treated mice. To this, STZ (80 mg/kg, i.p.) was administered to wild-type and STAT1(-/-) or IFN-gamma(-/-) mice for 5 days and the mice were euthanized after 14 days. STZ-induced beta-cell dysfunction and apoptosis were associated with increased STAT1/IRF-1 and ATF3 expression and were correlated with elevated IFN-gamma levels. Genetic depletion using IFN-gamma(-/-) or STAT1(-/-) mice strongly inhibited the reduction of islet cell mass or insulin synthesis/secretion and the increase of beta-cell apoptosis observed in STZ-treated wild-type mice. ATF3 overexpression, especially the C-terminal domain, strongly enhanced beta-cell dysfunction and apoptosis by enhancing STAT1 activation and its accumulation, which were abolished with an ATF3-specific siRNA or C-terminal-deleted ATF3. The STZ induction of ATF3 was completely depleted in IFN-gamma(-/-) mice, but not in STAT1(-/-) mice. Furthermore, STAT1 did not affect ATF3 expression, but STAT1 depletion or its inactivation inhibited STZ-induced ATF3 nuclear translocation and beta-cell apoptosis. Interestingly, ATF3 also increased STAT1 transcription by directly binding to a putative binding region (-116 to -96 bp) in the STAT1 promoter. Our results suggest that ATF3 functions as a potent upstream regulator of STAT1 and ATF3 may play a role in STZ-induced beta-cell dysfunction by enhancing the steady state abundance of STAT1.

Bio-composites comprised of kenaf fiber reinforced polylactide are fabricated by carding followed by treatment with a 3-glycidoxypropyl trimethoxy silane and hot-pressing. The effects of the silane coupling agent on composite properties was highly beneficial leading to increased moduli and heat deflection temperatures as well as reduced water swelling. Swelling is found to increase with increasing kenaf loading levels but to decrease with increasing coupling agent concentrations. Mechanical properties of the bio-composites at tempertaures above the glass transition are vastly improved in comparison to the base PLA polymer. As little as 10 wt.% kenaf fiber in PLA provides a heat distortion temperature in excess of 140 °C when combined with the coupling agent. The thermal stability towards degradation is slightly decreased relative to the base PLA and the ash content is increased. Thermal properties such as the glass transition, melting temperature, and percent crystallinity of the matrix PLA are largely unaffected by incorporation into the composites. Evidence of successful reaction of the silane with the kenaf fibers is provided by FTIR and implied by decreased swelling in water. A prototypical automotive headliner is fabricated as a means of demonstrating the viability of the present bio-composites in real manufacturing processes.

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Many receptors for endocytosis recycle into and out of cells through early endosomes. We now find in dendritic cells that the DEC-205 multilectin receptor targets late endosomes or lysosomes rich in major histocompatibility complex class II (MHC II) products, whereas the homologous macrophage mannose receptor (MMR), as expected, is found in more peripheral endosomes. To analyze this finding, the cytosolic tails of DEC-205 and MMR were fused to the external domain of the CD16 Fcγ receptor and studied in stable L cell transfectants. The two cytosolic domains each mediated rapid uptake of human immunoglobulin (Ig)G followed by recycling of intact CD16 to the cell surface. However, the DEC-205 tail recycled the CD16 through MHC II–positive late endosomal/lysosomal vacuoles and also mediated a 100-fold increase in antigen presentation. The mechanism of late endosomal targeting, which occurred in the absence of human IgG, involved two functional regions: a membrane-proximal region with a coated pit sequence for uptake, and a distal region with an EDE triad for the unusual deeper targeting. Therefore, the DEC-205 cytosolic domain mediates a new pathway of receptor-mediated endocytosis that entails efficient recycling through late endosomes and a greatly enhanced efficiency of antigen presentation to CD4+ T cells.

The protective effect of ginsenoside Re, isolated from ginseng berry, against acute gastric mucosal lesions was examined in rats with a single intraperitoneal injection of compound 48/80 (C48/80). Ginsenoside Re (20 mg/kg or 100 mg/kg) was orally administered 0.5 h prior to C48/80 treatment. Ginsenoside Re dose-dependently prevented gastric mucosal lesion development 3 h after C48/80 treatment. Increases in the activities of myeloperoxidase (MPO; an index of neutrophil infiltration) and xanthine oxidase (XO) and the content of thiobarbituric acid reactive substances (TBARS; an index of lipid peroxidation) and decreases in the contents of hexosamine (a marker of gastric mucus) and adherent mucus, which occurred in gastric mucosal tissues after C48/80 treatment, were significantly attenuated by ginsenoside Re. The elevation of Bax expression and the decrease in Bcl2 expression after C48/80 treatment were also attenuated by ginsenoside Re. Ginsenoside Re significantly attenuated all these changes 3 h after C48/80 treatment. These results indicate that orally administered ginsenoside Re protects against C48/80-induced acute gastric mucosal lesions in rats, possibly through its stimulatory action on gastric mucus synthesis and secretion, its inhibitory action on neutrophil infiltration, and enhanced lipid peroxidation in the gastric mucosal tissue.