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Particulate matter (PM) is an environmental hazard that is associated with various human health risks. The olfactory system is directly exposed to PM; therefore, the influence of PM exposure on olfactory function must be investigated. In this study, we propose a zebrafish olfactory model to evaluate the effects of exposure to diesel particulate matter (DPM), which was labeled Korean diesel particulate matter (KDP20). KDP20 comprises heavy metals and polycyclic aromatic hydrocarbons (PAHs). KDP20 exposed olfactory organs exhibited reduced cilia and damaged epithelium. Olfactory dysfunction was confirmed using an odor-mediated behavior test. Furthermore, the olfactory damage was analyzed using Alcian blue and anti-calretinin staining. KDP20 exposed olfactory organs exhibited histological damages, such as increased goblet cells, decreased cell density, and calretinin level. Quantitative real-time polymerase chain reaction (qRT-PCR) revealed that PAHs exposure related genes (AHR2 and CYP1A) were upregulated. Reactive oxidation stress (ROS) (CAT) and inflammation (IL-1B) related genes were upregulated. Furthermore, olfactory sensory neuron (OSN) related genes (OMP and S100) were downregulated. In conclusion, KDP20 exposure induced dysfunction of the olfactory system. Additionally, the zebrafish olfactory system exhibited a regenerative capacity with recovery conditions. Thus, this model may be used in future investigating PM-related diseases. [ABSTRACT FROM AUTHOR]

Retinal apoptosis plays a critical role in the progression of diabetic retinopathy (DR), a common diabetic complication. Currently, the tight control of blood glucose levels is the standard approach to prevent or delay the progression of DR. However, prevalence of DR among diabetic patients remains high. Focusing on natural nutrients or herbal medicines that can prevent or delay the onset of diabetic complications, we administered an ethanol extract of the aerial portion of Osteomeles schwerinae (OSSCE), a Chinese herbal medicine, over a period of 17 weeks to spontaneously diabetic Torii (SDT) rats. OSSCE was found to ameliorate retinal apoptosis through the regulation of advanced glycation end product (AGE) accumulation, oxidative stress, and mitochondrial function via the inhibition of NF-κB activity, in turn, through the downregulation of PKCδ, P47phox, and ERK1/2. We further demonstrated in 25 mM glucose-treated human retinal microvascular endothelial cells (HRMECs) that hyperoside (3-O-galactoside-quercetin), quercitrin (3-O-rhamnoside-quercetin), and 2″-O-acetylvitexin (8-C-(2″-O-acetyl-glucoside)-apigenin) were the active components of OSSCE that mediated its pharmacological action. Our results provide evidence that OSSCE is a powerful agent that may directly mediate a delay in the development or disease improvement in patients of DR. [ABSTRACT FROM AUTHOR]

Recently, real-time human age estimation based on facial images has been applied in various areas. Underneath this phenomenon lies an awareness that age estimation plays an important role in applying big data to target marketing for age groups, product demand surveys, consumer trend analysis, etc. However, in a real-world environment, various optical and motion blurring effects can occur. Such effects usually cause a problem in fully capturing facial features such as wrinkles, which are essential to age estimation, thereby degrading accuracy. Most of the previous studies on age estimation were conducted for input images almost free from blurring effect. To overcome this limitation, we propose the use of a deep ResNet-152 convolutional neural network for age estimation, which is robust to various optical and motion blurring effects of visible light camera sensors. We performed experiments with various optical and motion blurred images created from the park aging mind laboratory (PAL) and craniofacial longitudinal morphological face database (MORPH) databases, which are publicly available. According to the results, the proposed method exhibited better age estimation performance than the previous methods. [ABSTRACT FROM AUTHOR]

The root of Achyranthes bidentata Blume, Achyranthis Radix (AR), is used as a traditional medicine ingredient in East Asia. It has anti-inflammatory, anti-oxidative, and anti-diabetic activities. In the present study, we aimed to evaluate the oral toxicity and genotoxicity of single-dose and 4-week repeated-doses of AR hot water extract (ARE), under the good laboratory practice principles. For oral toxicity studies, SD rats (n = 5 per sex and group) were administered ARE at concentrations of 500, 1000, and 2000 mg/kg/day once (single dose) or once per day for 4 weeks (repeated dose). The non-clinical genotoxicity study consisted of bacterial reverse mutation using Escherichia coli (WP2 uvrA) and Salmonella typhimurium (TA98, TA100, TA1535, and TA1537), in vitro chromosomal aberration test with Chinese hamster lung cells (CHL/IU), and in vivo mouse bone marrow micronucleus test using bone marrow cells collected from male ICR mice (n = 5) that were orally administered ARE. In the single-dose oral toxicity study, mortality and treatment-related changes in body weight were not observed throughout the study, and the lethal dose was estimated to be > 2000 mg/kg in rats. In the 4-week repeated-dose oral toxicity study, ARE did not induce significant changes in body weight, organ weight, food intake, or hematological and serum biochemical parameters in any group. In the bacterial reverse mutation test, ARE did not induce gene mutations in any tested strain. In the chromosomal aberration test, ARE did not cause chromosomal aberrations. The micronucleus test showed no significant increase in the number of micronucleated polychromatic erythrocytes or the mean ratio of polychromatic to total erythrocytes. These results showed that ARE does not induce oral toxicity and genotoxicity in the in vivo and in vitro test systems. [Display omitted] [ABSTRACT FROM AUTHOR]

Background: The incidence of epistaxis during nasotracheal intubation via the left nostril is more frequent than that during intubation via the right nostril. This study evaluated the effect of the reverse bevel and tip direction of the nasotracheal tube on the incidence of epistaxis during nasotracheal intubation via the left nostril., Methods: Patients undergoing right-sided maxillofacial surgery requiring left nasotracheal intubation were randomly allocated to the control (tracheal tube in the conventional direction) or reverse (a 180˚ reverse direction, with the tube bevel facing the nasal septum and the leading edge (i.e., the tip) of the bevel pointing away from the nasal septum) groups (n = 37 for both). The primary outcome was the incidence of epistaxis evaluated using videolaryngoscopy., Results: The incidence of epistaxis in the reverse group was significantly lower than that in the control group (9 [24.3%] vs. 20 [54.1%], P = 0.009; relative risk: 0.45, 95% CI [0.24, 0.85], absolute risk reduction: 29.8%, number needed to treat: 3). The severity of epistaxis was significantly lower in the reverse group (P = 0.002). The first attempt nasal passage (P = 0.027) was significantly higher in the reverse group. Postoperative nasal pain was lower (P < 0.001), and patient satisfaction was higher (P < 0.001) in the reverse group. Nasotracheal tube-related complications did not occur in either group., Conclusions: The reverse bevel and tip direction of the nasotracheal tube reduced the incidence and severity of epistaxis and increased patient satisfaction among patients undergoing left nasotracheal intubation.

Background: Cervical epidural block (CEB) is an effective intervention for managing cervical radicular pain. This study aimed to investigate the current status of performing CEB in South Korea., Methods: Pain physicians affiliated with the Korean Pain Society were asked to complete anonymous questionnaires regarding CEB between September and October 2022. The questionnaire consisted of 24 questions assessing the current status and methods of CEB in detail., Results: Of the 198 surveys collected, 171 physicians (86.4%) reported performing CEB. Among those, the majority (94.7%) used fluoroscopy during the procedure. The paramedian interlaminar (IL) approach was the most preferred method (50.3%). Respondents performing fluoroscopic-guided IL CEB were categorized into two groups based on clinical experience: those with ≤10 years of experience (≤10-year group, n = 91) and those with >10 years of experience (>10-year group, n = 71). The proportion of physicians obtaining informed consent in the ≤10-year group and >10-year group was 50.5% and 56.3%, respectively. When entering the epidural space during IL CEB, the contralateral oblique view was the second most frequently used in both groups (≤10-year group, 42.9%; >10-year group, 29.6%). In targeting the upper cervical lesions (C3-4), the proportion of respondents who used an IL space higher than C6-7 was 17.6% in the ≤10-year group and 29.5% in the >10-year experience group., Conclusions: This study demonstrated variability in the CEB technique used by pain physicians in South Korea. The findings highlight the need for education on informed consent and techniques to enhance safety.

Introduction: Stenotic thoracic ligamentum flavum hypertrophy can cause leg and/or low back pain similar to that caused by lumbar spinal stenosis. However, the thoracic spine may occasionally be overlooked in patients with leg and/or low back pain. An accurate understanding of the prevalence of stenotic thoracic ligamentum flavum hypertrophy and its associated factors is necessary., Methods: In this prevalence study, we reviewed whole-spine MRI scans of patients who visited the pain clinic complaining of leg and/or low back pain between 2010 and 2019. We analyzed the overall prevalence and prevalence according to the age group, sex, grade of lumbar disc degeneration, and thoracic level. In addition, we identified factors independently associated with stenotic thoracic ligamentum flavum hypertrophy occurrence., Results: Among 1896 patients, the overall prevalence of stenotic thoracic ligamentum flavum hypertrophy was 9.8% (185/1896), with the highest prevalence observed in the ≥80-year-old age group among all age groups (15.9%, 14/88). The region with the highest prevalence was the T10/11 level (3.0%, 57/1896). Multivariable logistic regression analysis revealed that when compared with the <50-year-old age group, all other age groups were significantly associated with stenotic thoracic ligamentum flavum hypertrophy (p<0.01). In addition, grade 5 of lumbar disc degeneration was significantly associated with stenotic thoracic ligamentum flavum hypertrophy (p=0.03)., Conclusions: Given the possibility for missed stenotic thoracic ligamentum flavum hypertrophy to potentially result in neurological complications, extending lumbar spine MRI covering the lower thoracic region may be considered for patients over 50 years of age with suspected severe lumbar disc degeneration., Competing Interests: Competing interests: None declared., (© American Society of Regional Anesthesia & Pain Medicine 2024. No commercial re-use. See rights and permissions. Published by BMJ.)

Background: Although elective surgery for unruptured intracranial aneurysms (UIA) has increased, few studies have evaluated the risk factors for transfusion during UIA surgery. We evaluated the association between the preoperative De Ritis ratio (aspartate transaminase/alanine transaminase) and the incidence of intraoperative transfusion in patients who had undergone surgical UIA clipping., Methods: Patients who underwent surgical clipping of UIA were stratified into two groups according to the preoperative De Ritis ratio cutoff levels (< 1.54 and ≥ 1.54), and the propensity score (PS)-matching analysis was performed to compare the incidence of intraoperative transfusion. Logistic regression analyses were performed to determine the risk factors for intraoperative transfusion. Net reclassification improvement (NRI) and integrated discrimination improvement (IDI) analyses were performed to verify the improvement in the intraoperative transfusion predictive model upon addition of the De Ritis ratio., Results: Intraoperative transfusion incidence was 15.4% (77/502). We observed significant differences in the incidence of intraoperative transfusion (16.2% vs. 39.7%, P = 0.004) between the groups after matching. In the logistic regression analyses, the De Ritis ratio ≥ 1.54 was an independent risk factor for transfusion (odds ratio [OR]: 3.04, 95% CI [1.53, 6.03], P = 0.002). Preoperative hemoglobin (Hb) value was a risk factor for transfusion (OR: 0.33, 95% CI [0.24, 0.47], P < 0.001). NRI and IDI analyses showed that the De Ritis ratio improved the intraoperative blood transfusion predictive models (P = 0.031 and P = 0.049, respectively)., Conclusions: De Ritis ratio maybe a significant risk factor for intraoperative transfusion in UIA surgery.

Background: The preemptive visceral analgesic effect of regional nerve block has not been adequately investigated to date. We evaluated the preemptive visceral analgesic effect of thoracic paravertebral block (TPVB) in patients undergoing laparoscopic cholecystectomy (LC) in whom pre-incisional rectus sheath block (RSB) was used to minimize somatic surgical pain., Methods: In this prospective, randomized, assessor-blind study, 70 patients scheduled for elective LC were randomly assigned to the pre-TPVB (n = 35) or the post-TPVB (n = 35) group. Both groups received pre-incisional RSB, and patients in the pre-TPVB group received TPVB before skin incision while those in the post-TPVB group received TPVB after skin closure. The primary outcome was the total rescue analgesic consumption (morphine equianalgesic dose) during the 24 h post-surgery. The secondary outcomes were the cumulative analgesic consumption and pain intensity for 24 h after surgery, and adverse events., Results: Pre-TPVB significantly reduced total rescue analgesic consumption (estimated mean [95% CI]) during the 24 h after surgery than post-TPVB (16.9 [14.5, 19.3] vs. 25.3 [22.8, 27.7] mg, estimated difference: -8.3 [-11.8, -4.9], P < 0.001). The cumulative rescue analgesic consumption was significantly lower in the pre-TPVB group from 2-24 h after surgery (P < 0.001). The postoperative pain intensity was significantly lower in the pre-TPVB group as well at 0.5-6 h after surgery. There were no adverse events in both groups., Conclusions: Pre-incisional TPVB conferred a significant preemptive visceral analgesic effect in patients undergoing LC, and significantly reduced the amount of postoperative opioid consumption.

Background: Degenerative lumbar spondylolisthesis (DLS) is frequently associated with lumbar spinal stenosis (LSS) and conservative treatments such as epidural steroid injection do not have long-term benefits in LSS patients with DLS. This study evaluated the effectiveness of percutaneous epidural neuroplasty using a balloon catheter in patients with LSS and DLS., Methods: Patients' sex, age, body mass index, diabetes, hypertension, stenosis grading, pain duration, location, pain intensity, and medications were retrieved from electronic medical records. At 1, 3, and 6 months following the procedure, data on pain severity, medication usage, and physical functional status were analyzed. A generalized estimating equations model was used at the six-month follow-up. Patients were divided into those with DLS (the spondylolisthesis group) and those without DLS (the no spondylolisthesis group) to evaluate whether the effects of percutaneous epidural neuroplasty using a balloon catheter were different., Results: A total of 826 patients were included (spondylolisthesis: 433 patients, 52.4%; no spondylolisthesis: 393 patients, 47.6%). Age, body mass index, hypertension, pain location, and stenosis grading were statistically different between the two groups. The generalized estimating equations analyses with unadjusted and adjusted estimation revealed a significant improvement in the estimated mean numerical rating scale of pain intensities compared to that at baseline in both groups (P < 0.001). Any adverse events that occurred were minor and temporary., Conclusions: Percutaneous epidural neuroplasty using a balloon catheter may be an alternative treatment option for patients with chronic LSS, regardless of accompanying DLS, who have had failed conservative management.

Background: Cervical epidural steroid injection is associated with rare but potentially catastrophic complications. The contralateral oblique (CLO) view may be a safe and feasible alternative to the lateral (LAT) view for fluoroscopic-guided cervical epidural steroid injection. However, evidence for the clinical usefulness of the CLO view for cervical epidural steroid injection is lacking. We assessed the clinical usefulness of the CLO view for cervical epidural steroid injection in managing cervical herniated intervertebral discs., Methods: Patients were randomly assigned to receive fluoroscopic-guided cervical epidural steroid injection under LAT view or CLO view at 50±5° degrees groups. The primary outcome was the needling time comparison between the two groups. Secondary outcomes were comparison of first-attempt success rate, needle tip visualization and location, total number of needle passes, final success rate, crossover success rate and false-positive/negative loss of resistance. Complications and radiation dose were also compared., Results: The needling time significantly decreased in the CLO than in the LAT group. The first-attempt success rate was significantly higher in the CLO compared with the LAT group. The needle tip was clearly visualized (p<0.001) and located more often on (or just anterior to) the ventral interlaminar line (p<0.001) in the CLO than in the LAT group. There were significantly fewer needle passes (p=0.019) in the CLO than in the LAT group. There were no significant differences in the final success, crossover success, false-positive/negative loss of resistance or radiation dose between the groups. Two (5.9%) cases in the LAT group experienced complications., Conclusion: The CLO view may be recommended for fluoroscopic-guided cervical epidural steroid injection, considering its better clinical usefulness over the LAT view., Competing Interests: Competing interests: None declared., (© American Society of Regional Anesthesia & Pain Medicine 2022. No commercial re-use. See rights and permissions. Published by BMJ.)

EGHB010 is a standardized herbal formula of the rhizome mixture of Paeonia lactiflora Pallas and Glycyrrhiza uralensis Fisch. Neovascularization in the retina is a common pathophysiology of diabetic retinal microvasculopathy and exudative macular degeneration. In this study, we evaluated the inhibitory effects of EGHB010 on abnormal retinal angiogenesis in a hyperoxia-induced neovascular retinopathy model. Vascular endothelial growth factor (VEGF)-mediated vascular tube formation was assayed in human umbilical vascular endothelial cells (HUVECs). Experimental angiogenesis in the retinas was induced by exposing C57BL/6 pups to hyperoxic environment (75% oxygen) on postnatal day 7 (P7) and then returning them to normal oxygen pressure on P12. EGHB010 (50 and 100 mg/kg/day) was administered intraperitoneally for 5 days (P12 - P16). Retinal flat mounts were prepared to measure the extent of retinal neovascularization on P17. The incubation of HUVECs with EGHB010 (1-25 μg/mL) resulted in the inhibition of VEGF-mediated tube formation in a dose-dependent manner. EGHB010 at doses of 50 and 100 mg/kg/day inhibited the formation of retinal neovascular tufts by 31.15±2.28% and 59.83±2.92%, respectively. Together, our results indicate that EGHB010 is a potent anti-angiogenic agent and may have potential for the control of abnormal retinal vessel growth in patients with ischemic retinopathy.

Background: Diabetes mellitus (DM) is one of the most common diseases found across the world. Aster koraiensis extract (AKE) has a protective effect on diabetic complications such as diabetic retinopathy. However, the effects of AKE on hyperglycemia-linked impairment of wound healing during DM have not been elucidated. In this study, we investigated the effects of AKE on delayed wound healing induced by DM., Methods: DM was induced by intraperitoneal administration of streptozotocin (STZ; 75 mg/kg) to Sprague Dawley (SD) rats. Next, a wound was induced on the back of rats after administration of STZ. Further, AKE was prepared using an alcoholic extraction of A. koraiensis and orally administered daily for 18 days. Wound healing was evaluated using an i n vitro migration assay and measuring the wound area in vivo . Skin tissue thickness was evaluated using hematoxylin and eosin staining. Matrix metalloprotease (MMP) activity and expression were detected using zymography and immunohistochemistry., Results: AKE administration improved the delayed migration of keratinocytes in hyperglycemic animals. It also attenuated an increase in keratinocyte MMP-2/9 activity induced by hyperglycemia. AKE protected against DM-induced impaired wound healing in rats and prevented the degradation of skin tissue induced by DM. In addition, AKE attenuated DM-induced increase in MMP-2/9 expression in skin tissue., Conclusions: In conclusion, AKE may promote wound healing by re-epithelization via promotion of keratinocyte migration and by attenuating the disruption of the skin tissue layer via MMP-2/9 inhibition during hyperglycemia.

Preservation of adequate perfusion pressures to the graft is a main focus of intraoperative management during kidney transplantation. We undertook this study to investigate the incidence of the higher use of inotropes in kidney transplant recipients and identify the patient outcomes and preoperative and intraoperative variables related to this.We retrospectively analyzed 1053 patients who underwent kidney transplantation at Asan Medical Center between January 2006 and February 2012, stratified by their inotropic score ([dopamine] + [dobutamine] + [epinephrine × 100] + [norepinephrine × 100]) <7 versus ≥7, wherein all doses are expressed as μg/kg/min. We evaluated preoperative characteristics, hemodynamic parameters, and intraoperative variables as well as postoperative outcomes, such as length of hospital stay and 1-year rejection and mortality rate.Receiver-operating characteristic analysis was performed to determine inotropic score to predict 1-year mortality. An inotropic score of 7 had the best combined sensitivity and specificity. An inotropic score ≥7 (137 patients, 13.0%) was significantly more prevalent in older patients, those with polycystic kidney disease, and at a 2nd transplant. Anesthesia time, the amounts of crystalloid and 5% albumin infused, and the need for red blood cell transfusion were significantly higher in the inotropic score ≥7 group. The patients with a higher use of inotropes required longer postoperative hospital stay and experienced a >2-fold higher rejection within the 1st year and a 4-fold higher 1-year mortality rate.A higher use of inotropes in kidney transplant recipients is more prevalent in older patients, those with a 2nd transplant and in patients with polycystic kidney disease as their primary renal disease. The postoperative hospital stay, rejection within the 1st year, and 1-year mortality rate are increased in patients with an inotropic score ≥7., Competing Interests: The authors have no conflicts of interest to disclose.

Retinal pericyte loss and neovascularization are characteristic features of diabetic retinopathy. Gemigliptin, a dipeptidyl peptidase-4 (DPP-4) inhibitor, has shown robust blood-glucose lowering effects in type 2 diabetic patients, but its effects on diabetic retinopathy have not yet been reported. We evaluated the efficacy of gemigliptin on retinal vascular leakage in db/db mice, which is an animal model for type 2 diabetes, and neovascularization in oxygen-induced retinopathy (OIR) mice, which is an animal model for ischemic proliferative retinopathy. Gemigliptin (100mg/kg/day) was orally administered to the db/db mice for 12weeks. C57BL/6 mice on postnatal day 7 (P7) were exposed to 75% hyperoxia for 5days, followed by exposure to room air from P12 to P17 to induce OIR. Gemigliptin (50mg/kg/day) was intraperitoneally injected daily from P12 to P17. Retinal neovascularization was analyzed in flat-mounted retinas on P17. We determined the efficacy and possible mechanism of gemigliptin on high glucose-induced apoptosis of primary human retinal pericytes. The oral administration of gemigliptin for 4months significantly ameliorated retinal pericyte apoptosis and vascular leakage in the db/db mice. Gemigliptin also ameliorated retinal neovascularization in the OIR mice. Gemigliptin attenuated the overexpression of plasminogen activator inhibitor-1 (PAI-1) in the retinas of diabetic and OIR mice. Gemigliptin and PAI-1 siRNA significantly inhibited pericyte apoptosis by inhibiting the overexpression of PAI-1, which is induced by high glucose. Our results suggest that gemigliptin has potent anti-angiogenic and anti-apoptotic activities via suppressing DPP-4 and PAI-1, and the results support the direct retinoprotective action of gemigliptin., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Advanced glycation end products (AGEs) are involved in the development of diabetic complications such as diabetic retinopathy. 5'-methoxybiphenyl-3,4,3'-triol (referred to as K24) was isolated using bioactivity-guided fractionation of Osteomeles schwerinae C. K. Schneid. and identified as a potent AGE inhibitor. To identify the protective effect of K24 on disruption of the blood-retinal barrier, AGE-RSA was intravitreally injected into rat eyes. K24 had an inhibitory effect on AGE-RSA-induced retinal vascular leakage by suppressing the expression of vascular endothelial growth factor (VEGF) and decreasing the loss of occludin. In addition, we examined whether K24 has a preventive effect against retinal pathogenic angiogenesis in an oxygen-induced retinopathy (OIR) mouse model. K24 significantly reduced the retinal non-perfused area and neovascular tufts in the OIR mice. These data indicate that K24 could serve as an innovative pharmaceutical agent to prevent blood-retinal barrier breakage and retinal pathogenic angiogenesis through an anti-VEGF mechanism., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The retinal accumulation of advanced glycation end products (AGEs) is a condition, which is found in diabetic retinopathy. The purpose of the present study was to investigate the protective effect of Litsea japonica extract (LJE) and to elucidate its underlying protective mechanism in model diabetic db/db mice. Male, 7 -week-old db/db mice were treated with LJE (100 or 250 mg/kg body weight) once a day orally for 12 weeks. The expression levels of AGEs and their receptor (RAGE) were subsequently assessed by immunohistochemistry. An electrophoretic mobility shift assay and southwestern histochemistry were used to detect activated nuclear factor κB (NF-κB). The immunohistochemical analysis demonstrated that LJE significantly reduced the expression levels of the AGEs and RAGE in the neural retinas of the db/db mice. LJE markedly inhibited the apoptosis of retinal ganglion cells. In addition, LJE suppressed the activation of NF-κB. These results suggested that LJE may be beneficial for the treatment of diabetes-induced retinal neurodegeneration, and the ability of LJE to attenuate retinal ganglion cell loss may be mediated by inhibition of the accumulation of AGEs.

Advanced glycation end products (AGE) have been implicated in the development of diabetic retinopathy. Characterization of the early stages of diabetic retinopathy is retinal pericytes loss, which is the result of pericytes migration. In this study, we investigated the pathological mechanisms of AGE on the migration of retinal pericytes and confirmed the inhibitory effect of myricetin on migration in vitro and in vivo. Migration assays of bovine retinal pericytes (BRP) were induced using AGE-BSA and phosphorylation of Src, ERK1/2, focal adhesion kinase (FAK-1) and paxillin were determined using immunoblot analysis. Sprague-Dawley rats (6 weeks old) were injected intravitreally with AGE-BSA and morphological and immunohistochemical analysis of p-FAK-1 and p-paxillin were performed in the rat retina. Immunoblot analysis and siRNA transfection were used to study the molecular mechanism of myricetin on BRP migration. AGE-BSA increased BRP migration in a dose-dependent manner via receptor for AGEs (RAGE)-dependent activation of the Src kinase-ERK1/2 pathway. AGE-BSA-induced migration was inhibited by an ERK1/2 specific inhibitor (PD98059), but not by p38 and Jun N-terminal kinase inhibitors. AGE-BSA increased FAK-1 and paxillin phosphorylation in a dose- and time-dependent manner. These increases were attenuated by PD98059 and ERK1/2 siRNA. Phosphorylation of FAK-1 and paxillin was increased in response to AGE-BSA-induced migration of rat retinal pericytes. Myricetin strongly inhibited ERK1/2 phosphorylation and significantly suppressed pericytes migration in AGE-BSA-injected rats. Our results demonstrate that AGE-BSA participated in the pathophysiology of retinal pericytes migration likely through the RAGE-Src-ERK1/2-FAK-1-paxillin signaling pathway. Furthermore, myricetin suppressed phosphorylation of ERK 1/2-FAK-1-paxillin and inhibited pericytes migration., (Copyright © 2014 Elsevier Inc. All rights reserved.)

The accumulation of advanced glycation end products (AGEs) is associated with many of the complications of diabetes mellitus, including diabetic retinopathy. AGE-breakers, such as N-phenacylthiazolium and alagebrium, have been proposed as therapeutic agents for reversing the increase in protein crosslinking in diabetes. (-)-Epicatechin is a major dietary flavonoid with a wide range of health-promoting biological activities. The aim of this study was to determine the potential effect of (-)-epicatechin in reducing the burden of AGEs in vitro and in vivo and to evaluate whether the reduced AGE burden could translate into improvement in retinal vascular function in exogenously AGE-injected rats. Glycated human serum albumin was purified from patients with diabetes. The breakdown of the already formed AGEs was studied by treating glycated human serum albumin with (-)-epicatechin. To study the effect of (-)-epicatechin on retinal vascular function, exogenously AGE-injected rats were treated with (-)-epicatechin (50 and 100 mg/kg i.p.) for two weeks. Apoptosis of retinal vascular cells was quantified using TUNEL staining. The AGE load in the retinas was determined via immunohistochemical staining and western blot analysis. (-)-Epicatechin was able to break preformed glycated human serum albumin in vitro as well as reduce AGE accumulation in retinas in vivo in a dose dependent manner. In exogenously AGE-injected rats, treatment with (-)-epicatechin was evidenced by an improved retinal vascular apoptosis. AGE burden in retinas was also reduced upon treatment. This study suggests that (-)-epicatechin could represent a valuable drug for the treatment of diabetic retinopathy by reducing the AGE burden., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Aldose reductase (AR) is the first and rate-limiting enzyme of the polyol pathway. AR-dependent synthesis of excess polyols leads to lens opacification in diabetic cataract. The purpose of this study is to investigate the protective effect of Litsea japonica extract (LJE) on diabetes-induced lens opacification and its protective mechanism in db/db mice. Seven-week-old male db/db mice were treated with LJE (100 and 250 mg/kg body weight) once a day orally for 12 weeks. LJE dose dependently inhibited rat lens aldose reductase activity in vitro (IC50 = 13.53 ± 0.74 µg/mL). In db/db mice, lens was slightly opacified, and lens fiber cells were swollen and ruptured. In addition, lenticular sorbitol accumulation was increased in db/db mice. However, the administration of LJE inhibited these lenticular sorbitol accumulation and lens architectural changes in db/db mice. Our results suggest that LJE might be beneficial for the treatment of diabetes-induced lens opacification. The ability of LJE to suppress lenticular sorbitol accumulation may be mediated by the inhibition of AR activity.

Loss of blood-retinal barrier (BRB) properties is an important feature in the pathology of diabetic retinopathy. Endothelium integrity is important for the normal vascular function. Litsea japonica (Thunb.) Jussieu is a Korean native plant that is consumed as a vegetable food. In this study, we evaluated the ability of an ethanol extract of L. japonica to prevent retinal vascular leakages in db/db mice, which is an animal model of type II diabetes. L. japonica extracts (LJE, 100 and 250 mg/kg) were administered once a day, orally, for 12 weeks. Vehicle-treated db/db mice exhibited hyperglycemia and retinal vascular leakage. LJE treatment blocked diabetes-induced BRB breakdown and decreased retinal VEGF expression in db/db mice. LJE also inhibited the degradation of occludin, which is an important tight junction protein. These findings support the potential therapeutic usefulness of L. japonica for retinal vascular permeability diseases.

Chronic hyperglycemia leads to the formation of advanced glycation end products (AGEs), which accelerates the development of diabetic complications. Previous studies have shown that extract of Cassiae semen (CS), the seed of Cassia tora, has inhibitory activity on AGEs formation in vitro and reduces transforming growth factor-beta1 (TGF-β1) and extracellular matrix protein expression via inhibition of AGEs-mediated signaling in glomerular mesangial cells. In this study, to examine the preventive effects of CS extract on the development of diabetic nephropathy in vivo, streptozotocin (STZ)-injected diabetic rats were orally administered CS extract (200 mg/kg body weight/day) for 12 weeks. Serum glucose, triglycerides, and total cholesterol in diabetic rats were significantly higher compared to control rats. CS or aminoguanidine (AG) treatment significantly reduced these factors. Proteinuria and creatinine clearance were also significantly decreased in the CS-treated group compared with the untreated diabetic group. The CS-treated group had significantly inhibited COX-2 mRNA and protein, which mediates the symptoms of inflammation in the renal cortex of diabetic rats. Furthermore, histopathological studies of kidney tissue showed that in diabetic rats, AGEs, the receptor for AGEs, TGF-β1, and collagen IV were suppressed by CS treatment. Our data suggest that oral treatment of CS can inhibit the development of diabetic nephropathy via inhibition of AGEs accumulation in STZ-induced diabetic rats., (Copyright © 2013 Elsevier GmbH. All rights reserved.)

Increasing evidence indicates that advanced glycation end products (AGEs) contribute to the pathogenesis of diabetic nephropathy. The aim of this study was to investigate the protective effect of L. japonica extract (LJE) against renal damage in the db/db mouse. LJE (100 or 250 mg/kg per day) was given to diabetic mice for 12 weeks. Body weight, blood glucose levels, glycated hemoglobin (HbA1c) levels, and proteinuria were examined. In in vitro assay of the inhibition of AGE formation, immunohistochemical analysis of podocyte loss and AGE accumulations were performed. In 20-week-old db/db mice, severe hyperglycemia developed, and proteinuria was significantly increased. Diabetes induced markedly morphological alterations to the renal glomerular cells. AGE accumulations and podocyte loss were detected in renal glomeruli. LJE treatment significantly reduced proteinuria and AGE accumulations in diabetic mice. Moreover, the loss of nephrin, an important slit diaphragm component in the kidneys, was restored by LJE treatment. Our studies suggest that LJE might be beneficial for the treatment of diabetic nephropathy. The ability of LJE to attenuate proteinuria and podocyte dysfunction may be mediated by the inhibition of AGE accumulation in the context of diabetic nephropathy in db/db mice.

Retinal pericyte loss is one of the histopathological hallmarks of early diabetic retinopathy. Puerarin (4'-7-dihydroxy-8-beta-d-glucosylisoflavone), which is an isoflavone-C-glucoside, causes various pharmacological effects that include antihyperglycemic and anti-inflammatory activities. In the present study, we determined the efficacy and possible mechanism of puerarin on the advanced glycation end product (AGE)-modified bovine serum albumin (BSA)-induced apoptosis of cultured bovine retinal pericytes and rat retinal pericytes in intravitreally AGE-modified rat serum albumin (RSA)-injected eyes. Puerarin significantly inhibited pericyte apoptosis, the generation of reactive oxygen species (ROS), and NADPH oxidase activity by inhibiting the phosphorylation of p47phox and Rac1 which were induced by the AGE-BSA treatment. The puerarin treatment markedly suppressed the activation of nuclear factor-kappaB (NF-κB). In addition, the in vivo apoptosis of the retinal pericyte of rats that was stimulated by the intravitreal injection of AGE-RSA was evidently attenuated by the puerarin treatment. These results demonstrate that puerarin may exert inhibitory effects on AGE-induced pericyte apoptosis by interfering with the NADPH oxidase-related ROS pathways and blocking NF-κB activation, thereby ameliorating retinal microvascular dysfunction., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Betulinic acid is a pentacyclic triterpenic acid that exists naturally in many kinds of food and has many biological functions. The present study investigated the antiobesity properties of betulinic acid and possible mechanisms by which betulinic acid functions. To examine the antilipase function of betulinic acid, the ability of betulinic acid to inhibit pancreatic lipase activity in vitro and to prevent the elevation of plasma triacylglycerol levels was tested after oral administration of a lipid emulsion in rats. In addition, the lipolytic effects of betulinic acid were assayed in rat adipose tissues. The activity of cAMP-dependent phosphodiesterase was also measured in vitro. Betulinic acid inhibited pancreatic lipase activity in a dose-dependent manner at concentrations of 1.5-100 µM (IC₅₀ value of 21.10 µM) and prevented the elevation of plasma triacylglycerol levels 2 h after oral administration of the lipid emulsion at a dose of 100 mg/kg. In addition, betulinic acid had a strong lipolytic effect, which was mediated by cAMP-dependent phosphodiesterase inhibition. In conclusion, betulinic acid may exert antiobesity effects by directly inhibiting pancreatic lipase, which would prevent the absorption of lipid from the small intestine. In addition, it was found that betulinic acid may further accelerate fat mobilization by enhancing the levels of lipolysis in adipose tissues., (Copyright © 2011 John Wiley & Sons, Ltd.)

Background: One of the early signs of diabetic retinopathy is the alteration of the blood-retinal barrier (BRB), which may involve the breakdown of endothelial cell tight junctions. Methylglyoxal (MGO) is a cytotoxic metabolite that is produced from glycolysis in vivo. Elevated levels of MGO are observed in a number of pathological conditions, including neurodegenerative disorders and diabetic complications. Herein, we hypothesize that increased levels of MGO disrupt the tight junction protein known as occludin protein by matrix metalloproteinases (MMPs), leading to breakage of the BRB., Methods: MGO was intravitreally injected into eyes of rats. BRB leakage, MMPs activity, and occludin were investigated in intravitreally MGO-injected eyes., Results: When normoglycemic rats were intravitreally injected with 400 μM MGO, there was widespread leakage of fluorescein isothiocyanate-bovine serum albumin (FITC-BSA) from the retinal vasculature when compared to control retinas. In addition, MGO-injected retinas demonstrated increases of both activity and expression of MMP-2 and MMP-9, and the degradation of occludin was found in the MGO-injected retinas., Conclusions: The results suggest that the activation of MMPs by elevated levels of MGO in the retina may facilitate an increase in vascular permeability by a mechanism involving proteolytic degradation of occludin. These findings may have implications for the role of MGO in the pathogenesis of diabetic retinopathy.

To identify effective herb to treat obesity, we screened 115 herbal extracts for inhibition of porcine pancreatic lipase (triacylg-ycerol acylhydrolase, EC 3.1.1.3) activity in vitro. Of the extracts tested, Cudrania tricuspidata leaves exhibited the most pronounced inhibitory effect on lipase activity with an IC(50) value of 9.91 μg/mL. Antilipid absorption effects of C. tricuspidata leaves were examined in rats after oral administration of lipid emulsions containing 50 or 250 mg  C. tricuspidata/kg body weight. Plasma triacylglycerol levels 2 h after the oral administration of emulsions containing C. tricuspidata were significantly reduced compared to the untreated group (P < 0.05). These results suggest that C. tricuspidata leaves may be useful for the treatment of obesity.

Objectives: KIOM-79, a combination of four plant extracts, has a preventive effect on diabetic nephropathy and retinopathy in diabetic animal models. In this study, we have investigated the inhibitory effects of KIOM-79 on diabetic cataractogenesis., Methods: We evaluated aldose reductase activity during cataractogenesis using Zucker diabetic fatty (ZDF) rat, an animal model of type 2 diabetes. ZDF rats were treated orally with KIOM-79 (50 mg/kg body weight) once a day for 13 weeks., Key Findings: In vehicle-treated ZDF rats, lens opacity was increased, and lens fibre swelling and membrane rupture were observed. In addition, aldose reductase activity and aldose reductase protein expression in diabetic lens were markedly enhanced. However, the administration of KIOM-79 inhibited the development of diabetic cataract through the inhibition of aldose reductase activity and protein expression in diabetic lenses., Conclusions: These observations suggested that KIOM-79 was useful against the treatment of diabetic cataractogenesis., (© 2011 The Authors. JPP © 2011 Royal Pharmaceutical Society.)

Chlorogenic acid (5-O-caffeoylquinic acid, CA), a phenolic compound found ubiquitously in plants, has antidiabetic effect in diabetic animal models. In this study, we investigated the inhibitory effect of CA on diabetic cataractogenesis. We evaluated the aldose reductase (AR) activity during cataract development in 50% galactose-fed rats, an animal model of sugar cataract. Galactose-fed rats were treated orally with CA (10 and 50 mg/kg body weight) once a day for 2 weeks. In vehicle-treated galactose-fed rats, lens opacity was increased, and lens fiber swelling and membrane rupture were observed. In addition, AR protein was highly expressed in lens epithelial cells and lens cortical fibers of galactose-fed rats. However, CA inhibited the rat AR activity in vitro, and the administration of CA prevented the development of sugar cataract through the inhibition of AR activity. These observations suggest that CA is useful for the treatment of sugar cataract.

Background: The purpose of the experiment reported here was to assess the involvement of advanced glycation end products (AGEs), oxidative stress, and nuclear factor kappa-B (NF-κB) activation in the development of diabetic keratopathy., Methods: Diabetes was induced by intraperitoneal streptozotocin injection in male Sprague-Dawley rats. The thickness of the cornea was measured. Apoptosis was detected by TUNEL assay and western blot for caspase-3. The expression of AGEs and 8-hydroxydeoxyguanosine (8-OHdG) were studied by immunohistochemistry in corneal tissues of normoglycaemic and diabetic rats. NF-κB activation was evaluated by electrophoretic mobility shift assay and southwestern histochemistry., Results: Corneal edema was observed in diabetic rats. The thickness of cornea was higher in diabetic than in control rats. AGEs were accumulated in corneal tissues. 8-OHdG and NF-κB were identified in corneal epithelium, stroma and endothelium, and its expressions were greater in diabetic than in those of control rats. Diabetes induces significant alterations in rat corneal tissue structure., Conclusions: The higher expression of AGE, 8-OHdG and NF-κB in corneal tissues of diabetic rats suggests that these factors are involved in apoptosis and in subsequent corneal alterations related to diabetic keratopathy.

Objectives: KIOM-79 is a mixture of 80% ethanol extracts of parched Puerariae radix, gingered Magnoliae cortex, Glycyrrhizae radix and Euphorbiae radix. The preventive effect of KIOM-79 on the development of diabetic keratopathy has been investigated., Methods: Seven-week-old male Zucker diabetic fatty (ZDF) rats were treated with KIOM-79 (50 mg/kg body weight) once a day orally for 13 weeks. The thickness of the cornea was measured and the extent of corneal cell death was detected by a terminal deoxynucleotidyl transferase dUTP nick-end labelling assay. The expression of advanced glycation end products (AGEs), 8-hydroxydeoxyguanosine, nuclear factor-kappaB (NF-κB), Bax and Bcl-2 were evaluated in corneal tissues., Key Findings: The administration of KIOM-79 prevented corneal oedema and apoptotic cell death of corneal cells. The accumulation of AGE in corneal tissues was reduced in ZDF rats treated with KIOM-79. Moreover, KIOM-79 attenuated oxidative DNA damage, NF-κB activation and Bax overexpression in the cornea., Conclusions: The results suggested that KIOM-79 exhibited corneal protective properties by not only reducing oxidative stress but inhibiting the AGEs/NF-κB downstream signal pathway during the development of diabetic keratopathy., (© 2011 The Authors. JPP © 2011 Royal Pharmaceutical Society.)

Advanced glycation end products (AGEs) play an important role in the development of chronic diabetic complications. Chlorogenic acid (CGA) is a phenolic compound formed by the esterification of caffeic and quinic acids. In this study, we evaluated the inhibitory effects of CGA against the formation of AGEs and AGEs protein cross-linking in vitro. An in vitro assay for glycation of bovine serum albumin by high glucose showed that CGA inhibited AGEs formation with an IC(50) value of 148.32 μM and was found to be more effective than aminoguanidine, a well-known AGEs inhibitor (IC(50); 807.67 μM). In an indirect AGE-ELISA assay, the CGA exhibited more potent inhibitory activity on the cross-linking of AGEs to collagen than aminoguanidine. In addition, the inhibitory effects of CGA on AGEs formation and on its cross-linking with collagen might be caused by its interactions with reactive decarbonyl compounds, such as methylglyoxal. These results suggest that CGA could be beneficial in the prevention of AGEs progression in patients with diabetes because CGA can attenuate AGEs deposition in glucose.

Methylglyoxal (MGO), a cytotoxic metabolite, is produced from glycolysis. Elevated levels of MGO are observed in a number of diabetic complications, including retinopathy, nephropathy and cardiomyopathy. Loss of retinal pericyte, a hallmark of early diabetic retinal changes, leads to the development of formation of microaneurysms, retinal hemorrhages and neovasculization. Herein, we evaluated the cytotoxic role of MGO in retinal pericytes and further investigated the signaling pathway leading to cell death. Rat primary retinal pericytes were exposed to 400muM MGO for 6h. Retinal vessels were prepared from intravitreally MGO-injected rat eyes. We demonstrated apoptosis, nuclear factor-kappaB (NF-kappaB) activation and inducible nitric oxide synthase (iNOS) induction in cultured pericytes treated with MGO and MGO-injected retinal vessels. In MGO-treated pericytes, TUNEL-positive nuclei were markedly increased, and NF-kappaB was translocalized into the nuclei of pericytes, which paralleled the expression of iNOS. The treatment of pyrrolidine dithiocarbamate (an NF-kappaB inhibitor) or l-N6-(1-iminoethyl)-lysine (an iNOS inhibitor) prevented apoptosis of MGO-treated pericytes. In addition, in intravitreally MGO-injected rat eyes, TUNEL and caspase-3-positive pericytes were significantly increased, and activated NF-kappaB and iNOS were highly expressed. These results suggest that the increased expression of NF-kappaB and iNOS caused by MGO is involved in rat retinal pericyte apoptosis., (Copyright 2010 Elsevier Ireland Ltd. All rights reserved.)

Background: It has been suggested that damage of lens epithelial cell (LEC) may play an important role in cataract formation. Nitric oxide is involved in cataract development. Here, we investigated the relationship between LEC damage and iNOS expression in the Zucker diabetic fatty (ZDF) rat., Methods: At 21 weeks of age, the eyes were enucleated and the lens opacity was then examined. Apoptosis were detected by TUNEL assay, and the expression of iNOS and NF-kappaB activation were studied by immunohistochemistry and southwestern histochemistry respectively., Results: In 21-week-old male ZDF rats, cataract was developed, TUNEL-positive LECs were markedly increased, and the expression levels of iNOS mRNA and protein were significantly upregulated. The expression pattern of iNOS was closely correlated with apoptotic change of LECs. In addition, advanced glycation end products (AGEs) were accumulated in cytoplasm of LECs. Activated NF-kappaB was mainly detected in nucleus of LECs., Conclusions: The higher expressions of AGEs, NF-kappaB and iNOS in LECs of diabetic rats suggest that these factors are involved in apoptosis of LEC alterations related to diabetic cataract.

Four flavan-3-ols, (+)-catechin ( 1), (+)-catechin 7- O- beta- D-apiofuranoside ( 2), (+)-catechin 7- O- beta- D-xylopyranoside ( 3), (+)-catechin 7- O- beta- D-glucopyranoside ( 4), and proanthocyanidin A-1 ( 5) as well as three other constituents, isolated from an EtOAc-soluble extract of the stem barks of Ulmus davidiana var. japonica, were evaluated for inhibitory activity against the formation of AGEs. Compounds 1 - 5 exhibited a significant inhibitory activity on the formation of AGEs in an AGEs-BSA assay by specific fluorescence and this was confirmed by an indirect AGEs-ELISA. Moreover, compounds 1 and 5 markedly reduced AGEs-BSA cross-linking to collagen in a dose-dependent manner. AGEs:advanced glycation end products BSA:bovine serum albumin CC:column chromatography CD:circular dichroism.

We investigated the effect of magnolol (5,5'-diallyl-2,2'-dihydroxybiphenyl), a marker compound isolated from the cortex of Magnolia officinalis, in non-obese type 2 diabetic Goto-Kakizaki (GK) rats. The rats were treated orally with magnolol (100 mg/kg body weight) once a day for 13 weeks. In magnolol-treated GK rats, fasting blood glucose and plasma insulin were significantly decreased, and the pancreatic islets also showed strong insulin antigen positivity. Urinary protein and creatinine clearance (Ccr) were significantly decreased. Pathological examination revealed the prevention of the glomeruli enlargement in magnolol-treated GK rats. The overproduction of renal sorbitol, advanced glycation endproducts (AGEs), type IV collagen, and TGF-beta1 mRNA were significantly reduced in magnolol-treated GK rats. Thus based on our findings, the use of magnolol could result in good blood glucose control and prevent or retard development of diabetic complications such as diabetic nephropathy.

Cadmium (Cd), a ubiquitous heavy metal, has been shown to accumulate in the central nervous system, especially outside of the blood-brain barrier (BBB), suggesting a potential toxicity to nervous tissue. Thus, we investigated the effect of Cd on intercellular adhesion molecule-1 (ICAM-1) expression, as an indicator of BBB injury, in mouse brain microvessel endothelial cells (bEnd.3 cells). The treatment with Cd increased the expression of ICAM-1 at the levels of protein and mRNA, and these increases were almost completely inhibited by a specific NF-kappaB inhibitor SN50. The treatment with Cd induced the translocation of NF-kappaB from cytosolic to membrane fraction and increased DNA binding activity of NF-kappaB, and this NF-kappaB activation was inhibited by SN50. Interestingly, Cd did not trigger the degradation of IkappaBalpha, suggesting that Cd-induced ICAM-1 expression is mediated through IkappaBalpha degradation-independent pathway. Instead, tyrosine phosphorylation of IkappaBalpha was significantly elevated by Cd treatment, and this elevation was blocked by genistein, a protein tyrosine kinase inhibitor. In summary, the present results suggest that Cd stimulates the expression of ICAM-1 in bEnd.3 cells, via NF-kappaB activation that is mediated by the tyrosine phosphorylation of IkappaBalpha.

In this study, we investigated whether carbofuran, a commonly used carbamate pesticide, and N-nitrosocarbofuran (NOCF), the N-nitroso metabolite of carbofuran, have cytotoxicity in mouse brain microvascular endothelial cells (bEnd.3). Results from the MTT assay in bEnd.3 cells showed that NOCF but not carbofuran caused a remarkable decrease in cell viability. The cell death induced by NOCF appeared to involve apoptosis, based on our results from annexin V staining and electron microscopy. To investigate the mechanism of the NOCF-induced cell death, we examined the effects of selective inhibitors for MAP kinase pathways, PD98059 (for MEK/ERK), SB202190 (for p38 MAP kinase), and SP600125 (for JNK), on the NOCF-induced cell death. The NOCF-induced cell death was significantly reduced by PD98059, but not by SB202190 or SP600125. NOCF increased ERK phosphorylation as early as 15 min after the treatment and this increase was maintained for 2 h. In summary, our results suggest that NOCF can induce apoptotic cell death, at least in part, through the ERK pathway in brain microvascular endothelial cells.