One-Step Detection of Vancomycin in Whole Blood Using the Lateral Flow Immunoassay.

Vancomycin (VAN) is an effective antibiotic against Gram-positive bacteria and the first-line therapy to prevent and treat methicillin-resistant Staphylococcus aureus (MRSA) and severe infections. However, low concentrations of VAN can result in resistant strains. High doses of VAN can cause nephrotoxicity and ototoxicity; thus, VAN is a representative drug for which drug monitoring is recommended. Several methods have been proposed to detect VAN. Among them, lateral flow immunoassays (LFIAs) have advantages, such as simple and user-friendly operation, low sample volume requirement, and cost effectiveness. In this study, we developed an LFIA capable of rapid on-site detection such that the VAN concentration in plasma could be monitored within 20 min by a one-step detection process using whole blood without plasma separation. VAN can be detected in whole blood over a wide range of concentrations (20-10,000 ng/mL), and the LFIA reported here has a detection limit of 18 ng/mL. The applicability of the developed LFIA compared to the results of measuring VAN with a commercial enzyme-linked immunosorbent assay kit showed a satisfactory correlation (Spearman's rho, ρ = 0.891). Therefore, the developed LFIA enables rapid and wide-range VAN detection in whole blood and can aid in drug monitoring to evaluate patients' responses to treatment.

Evaluation of Vertical Level Differences Between Left and Right Vocal Folds Using Artificial Intelligence System in Excised Canine Larynx.

OBJECTIVES: This study aimed to establish an artificial intelligence (AI) system to classify vertical level differences between vocal folds during vocalization and to evaluate the accuracy of the classification. METHODS: We designed models with different depths between the right and left vocal folds using an excised canine larynx. Video files for the data set were obtained using a high-speed camera system and a color complementary metal oxide semiconductor camera with global shutter. The data sets were divided into training, validation, and testing. We used 20,000 images for building the model and 8000 images for testing. To perform deep learning multiclass classification and to estimate the vertical level difference, we introduced DenseNet121-ConvLSTM. RESULTS: The model was trained several times using different numbers of epochs. We achieved the most optimal results at 100 epochs, and the batch size used during training was 16. The proposed DenseNet121-ConvLSTM model achieved classification accuracies of 99.5% and 88.0% for training and testing, respectively. After verification using an external data set, the overall accuracy, precision, recall, and f1-score were 90.8%, 91.6%, 90.9%, and 91.2%, respectively. CONCLUSIONS: The newly developed AI system may be an easy and accurate method for classifying superior and inferior vertical level differences between vocal folds. Thus, this AI system can be applied and may help in the assessment of vertical level differences in patients with unilateral vocal fold paralysis.

Analgesic effects and metabolome analyses of laser- and electro-acupuncture combined therapies in paclitaxel-induced neuropathic pain model.

Introduction: Allodynia, which can be induced by paclitaxel administration, is the presence of pain as a result of a stimulus that does not usually provoke pain. Many studies have investigated the analgesic efficacy of acupuncture, including laser acupuncture (LA) and electroacupuncture (EA). Although pain-related diseases are relatively common, few studies have analyzed the analgesic effects and mechanisms of LA combined with EA. The purpose of this study was to investigate the therapeutic effect and mechanism of manual acupuncture (MA), EA, LA, and combined therapy (LA + EA) in a paclitaxel-induced allodynia rat model. Methods: A total of 56 rats were classified into eight groups: a normal (Nor, n = 7), a control (Con, n = 7), an MA (n = 7), an EA (n = 7), a 650-nm LA (650LA, n = 7), an 830-nm LA (830LA, n = 7), a 650-nm LA combined with EA (650LA + EA, n = 7), and an 830-nm LA combined with EA group (830LA + EA, n = 7). Allodynia was induced by intraperitoneal injection of 2 mg/kg of paclitaxel every other day for a total of four times except the Nor group. Acupuncture treatments were conducted at the points of Jungwan (CV12) and Joksamni (ST36) once every other day for 6 min, for a total of nine times. Withdrawal response reaction times and force intensity of the foot were measured before the start of the experiment, after the 4th paclitaxel administration (day 8), and after the 9th and last treatment (day 15). On the 16th day, mRNA and protein expression in the spinal nerves was assessed, and a metabolome analysis of the animals' feces was performed. Results and discussion: Our analyses show that 650LA + EA treatment resulted in an upregulation of protein expression related to pain relief and nerve regeneration, whereas 830LA + EA treatment led to significant changes in metabolomes. This study demonstrates that a combination treatment of EA and LA can suppress allodynia and promote upregulation of protein expression related to nerve regeneration and is effective in changing the intestinal microbiome. Further large-scale research is required to assess the exact mechanism underlying the therapeutic effect of this combination treatment in pain-related diseases.

A 13-Week Repeated Oral Dose Toxicity Study of ChondroT in Sprague-Dawley Rats.

BACKGROUND: ChondroT, a new herbal medication, consists of Angelica grosseserrata Maxim., Lonicera japonica Thunb., Angelica gigas Nakai, Clematis terniflora var. manshurica (Rupr.) Ohwi, and Phellodendron amurense Rupr. (6:4:4:4:3). Our previous studies have shown that ChondroT exhibits significant anti-arthritic and anti-inflammatory effects. In this study, we aimed to assess the toxicological safety assessment of ChondroT. METHODS: This study was designed to assess the safety of ChondroT after repeated oral administration. Male and female Sprague-Dawley rats were treated with ChondroT at oral doses of 0, 500, 1000, and 2000 mg/kg for 13 weeks. Mortality, clinical signs, body weight changes, food consumption, ophthalmological findings, urinalysis, hematological and blood-chemical parameters, necropsy findings, organ weights, and histological markers were recorded throughout the study period. Rats were also monitored for an additional 4 weeks to determine the recovery time. RESULTS: No death occurred and no significant changes in food consumption, ophthalmologic findings, and urinalysis were found. Although there were alterations in clinical signs, body weights, hematological parameters, blood-chemical parameters, necropsy findings, organ weights, and histological markers, they were not considered to be toxicologically significant. CONCLUSIONS: The results suggest that the no-observed adverse effects level (NOAEL) was 2000 mg/kg/day for the test substance. ChondroT, a new complex herbal medication composed of five plants, can therefore be used safely at the NOAEL.

Inhibitory effects of ChondroT and its constituent herbs on RANKL-induced osteoclastogenesis.

BACKGROUND: ChondroT is a complex herbal medicine consisting of water extracts of Ostericum koreanum (Maxim.) Kitag., Lonicera japonica Thunb., Angelica gigas Nakai, Clematis manshurica Rupr., and Phellodendron amurense Rupr. (6:4:4:4:3). Previous studies have reported that ChondroT possesses chondroprotective and anti-inflammatory, anti-osteoarthritic, and anti-hyperuricemic activities. The study is aim to demonstrate the effects of ChondroT and its five constituent herbs on receptor activator of NF-κB ligand (RANKL)-induced osteoclastogenesis and the underlying mechanisms. METHODS: Osteoclastogenesis was identified in bone marrow-derived macrophages (BMDMs) by tartrate-resistant acid phosphatase (TRAP) staining assay, actin ring formation assay and the bone resorption assay. For the molecular mechanisms, activation of RANKL-induced NF-κB and MAPK signaling pathways and the expression levels of osteoclast-specific proteins were investigated by Western blotting. Cell viability was assessed by MTT assay. Actin ring formation and NF-κB translocation were evaluated by immunostaining. RESULTS: ChondroT and each of its constituent herbs significantly suppressed osteoclast differentiation dose dependently, and decreased actin ring formation as well as bone-resorbing capacity. Mechanistically, ChondroT and its constituent herbs downregulated the expressional levels of osteoclast-specific proteins such as NFATc1, c-Fos, Cathepsin K, and matrix metalloproteinase 9 (MMP9) by suppressing NF-κB translocation to nucleus and MAPKs phosphorylation at different levels. Compared to its five constituent herbs, ChondroT exhibited the best inhibitory efficiency against osteoclastogenesis. CONCLUSIONS: Taken together, ChondroT has anti-osteoclastogenesis properties by inhibiting NF-κB and MAPKs pathways. It could be considered as a potential therapeutic candidate for the treatment of osteoclast-related bone diseases.

Effects of ChondroT on potassium Oxonate-induced Hyperuricemic mice: downregulation of xanthine oxidase and urate transporter 1.

BACKGROUND: ChondroT, a new herbal medication, consists of the water extracts of Osterici Radix, Lonicerae Folium, Angelicae Gigantis Radix, Clematidis Radix, and Phellodendri Cortex (6:4:4:4:3). We previously reported that ChondroT showed significant anti-arthritis and anti-inflammatory effects. METHODS: This study was designed to evaluate the effect of ChondroT on hyperuricemia. First, the effect of ChondroT was evaluated on xanthine oxidase (XOD) activity in vitro. The anti-hyperuricemic effect of ChondroT was also studied in potassium oxonate (PO)-induced hyperuricemic model mice. Uric acid (UA) and XOD were evaluated in the serum, urine, and liver of the mice. In addition, we measured serum creatinine (Cr) and blood urea nitrogen (BUN) levels as well as mRNA expression of the mouse urate transporter 1 (mURAT1) to evaluate kidney function and urate excretion in hyperuricemic mice. RESULTS: ChondroT showed in vitro XOD inhibitory activity in a dose-dependent manner (P < 0.05). We demonstrated that ChondroT (37.5, 75 and 150 mg/kg) significantly reduced serum UA (P < 0.01 and P < 0.001, respectively), and upregulated urinary UA (P < 0.001, respectively) in PO-induced hyperuricemic mice. In addition, ChondroT (75 and 150 mg/kg) significantly reduced Cr (P < 0.05 and P < 0.01, respectively), BUN (P < 0.05 and P < 0.001, respectively), GOT (P < 0.05 and P < 0.01, respectively), and GPT (P > 0.05 and P < 0.05, respectively) levels in PO-induced hyperuricemic mice. ChondroT (75 and 150 mg/kg) also significantly downregulated serum (P < 0.05) and liver (P < 0.05) XOD activity. Compared to the hyperuricemic mice, the ChondroT (37.5, 75, and 150 mg/kg)-treated mice showed decreased mURAT1 protein expression level. CONCLUSION: ChondroT displayed anti-hyperuricemic effects by regulating XOD activity and kidney mURAT1.

Anti-osteoarthritic effects of ChondroT in a rat model of collagenase-induced osteoarthritis.

BACKGROUND: Previously, we reported that ChondorT showed significant anti-arthritis and anti-inflammatory effects. ChondroT, a new herbal medication, consists of the water extracts of Osterici Radix, Lonicerae Folium, Angelicae Gigantis Radix, Clematidis Radix, and Phellodendri Cortex (6:4:4:4:3). The objective of this study was to investigate the effects of ChondroT in collagenase-induced osteoarthritis rat model. METHODS: Osteoarthritis was induced by the injection of collagenase into the right knee joint cavity of rats. The samples were divided into seven groups [intact (n = 6), control (n = 6), indomethacin (n = 6), Joins tab (n = 6), ChondroT50 (n = 6), ChondroT100 (n = 6), and ChondroT200 (n = 6)]. The control group was administered normal saline, indomethacin group was administered indomethacin (2 mg/kg), and Joins tab group was administered Joins Tab (20 mg/kg). The ChondroT50, ChondroT100, and ChondroT200 groups were administered 50, 100, and 200 mg/kg of ChondroT, respectively. All oral administrations were initiated 7 days after the induction of arthritis and were continued for a total of 12 days. At the end of the experiment, serum aminotransferase, albumin, blood urea nitrogen, creatinine, leukocyte, and inflammatory cytokines [tumor necrosis factor (TNF)-α, interleukin (IL)-1ß, and IL-6] were analyzed. Hematoxylin and eosin (H&E) and safranin O-fast green staining of the articular structures of the knee joint were performed. RESULTS: TNF-α and IL-1ß decreased in the ChondroT100 and ChondroT200 groups compared with those in the control group. IL-6 and aspartate aminotransferase decreased in the ChondroT50, ChondroT100, and ChondroT200 groups compared with that in the control group. Albumin, WBC and lymphocytes decreased in the ChondroT100 and ChondroT200 groups compared with those in the control group. In H&E stain, synoviocytes, cartilage lacunae, and chondrocytes were well preserved in the ChondroT100 and ChondroT200 groups, and safranin O-fast staining showed a clear reaction of proteoglycans in the ChondroT100 and ChondroT200 groups. CONCLUSIONS: Based on these results, it can be proposed that ChondroT has anti-osteoarthritic effects on collagenase-induced rat model.

Antiosteoarthritic Effects of ChondroT in a Rat Model of Monosodium Iodoacetate-Induced Osteoarthritis.

Ganghwaljetongyeum is a traditional Korean herbal medicine used to treat joint pain, limited motion, fever, and swelling; it also inhibits inflammatory processes associated with arthritis. ChondroT, a water extract of Ganghwaljetongyeum, is a new complex herbal medicine. This study investigated the effects of ChondroT using a rat model of monosodium iodoacetate- (MIA-) induced osteoarthritis. Thirty-six rats were randomly divided into three ChondroT groups and a normal, control, and positive control group. Changes in paw edema volume, histopathology, and plantar withdrawal response were analyzed. Further, inflammatory cytokines, arachidonic acids, liver and kidney function, and hematological features were measured. ChondroT significantly decreased paw edema by the 5th day and notably improved articular cartilage damage; it also significantly improved the plantar withdrawal response in terms of both reaction time and force intensity. Moreover, treatment with ChondroT significantly decreased the serum levels of tumor necrosis factor alpha, interleukin-1ß, interleukin-6, and prostaglandin E2 and significantly increased serum aspartate aminotransferase and alanine aminotransferase levels. This study demonstrates that ChondroT has anti-inflammatory and analgesic effects in a MIA-induced osteoarthritis rat model. These results support the clinical relevance of ChondroT for future use in patients with osteoarthritis. However, further studies are required to elucidate the corresponding mechanisms.

Efficacy and safety of ChondroT on knee-osteoarthritis: Protocol for a 8-week, randomized, double-blind, placebo-controlled, multicenter therapeutic exploratory clinical trial.

BACKGROUND: Arthritis is the most common disease in elderly individuals. Many medications for osteoarthritis treatment have the potential for side effects. Using a bioinformatics tool and preclinical studies, ChondroT, 5 herbal complexes, was identified from Ganghwaljetongyeum, which is a18 herbal complex, which has often used to treat osteoarthritis. The goal of this study is to evaluate short-term safety of ChondroT. METHODS: This will be a multicenter, randomized, double-blind, and placebo-controlled trial. There will be a 2-week run-in period before random allocation to 3 groups, ChondroT 1.0 g, 2.0 g, and placebo groups. Total duration of the clinical trial will be 14 weeks including a 4-week washout follow-up. Participants will be followed-up every 4 weeks, and the effect and the safety will be assessed at visit 2, 3, and 4. All participants are asked to maintain the medication schedule in this protocol. The primary outcome will be measured using pain visual analog scale (VAS) after 8 week treatment and secondary outcomes will include pain VAS score after 4 week treatment, SF-36 survey score, patient's global assessment, physical function test, and the change of erythrocyte sedimentation rate, and C-reactive protein. The repeated-measure analysis will be used for the primary efficacy based on full analysis set and per-protocol. DISCUSSION: This study has restrictive inclusion, exclusion criteria, and a well-controlled intervention, and it will be the first randomized controlled trial to evaluate the efficacy and safety of ChondroT formula in osteoarthritis patients. The trial according to this protocol may provide a new intervention in the treatment of osteoarthritis.

Real-time Simultaneous DKG and 2D DKG Using High-speed Digital Camera.

INTRODUCTION: For the evaluation of voice disorders, direct observation of vocal cord vibration is important. Among the various methods, laryngeal videostroboscopy (LVS) is widely used, but it was not a true image because it collects images from different cycles. In contrast, high-speed videoendoscopy and videokymography have much higher frame rates and can assess functional and mobility disorders. OBJECTIVE: The purpose of the study is to describe real-time, simultaneous digital kymography (DKG), two-dimensional scanning (2D) DKG, and multi-frame (MF) LVS system using a high-speed digital camera, and identify the efficacy of this system in evaluating vibratory patterns of pathologic voice. METHODS: The pattern of vocal fold vibration was evaluated in a vocally healthy subject and in subjects with vocal polyp, vocal nodules, vocal cord scar, and vocal cord paralysis. We used both quantitative (left-right phase symmetry, amplitude symmetry index) and qualitative (anterior-posterior phase symmetry) parameters for assessment of vocal fold vibration. RESULTS: Our system could record videos within seconds and required relatively little memory. The speed of replay in the DKG, 2D DKG, MF LVS, and high-speed videoendoscopy was controllable. The number of frame per cycle with MF LVS was almost the same as the fundamental frequency. CONCLUSION: Our system can provide images of various modalities simultaneously in real time and analyze morphological and functional vibratory patterns. It can be possible to provide a greater level of information for the diagnosis and treatment of vibratory disorders.

Chondroprotective and anti-inflammatory effects of ChondroT, a new complex herbal medication.

BACKGROUND: Ganghwaljetongyeum (GHJTY) is a complex herbal decoction comprising 18 plants; it is used to treat arthritis. In order to develop a new anti-arthritic herbal medication, we selected 5 out of 18 GHJTY plants by using bioinformatics analysis. The new medication, called ChondroT, comprised water extracts of Osterici Radix, Lonicerae Folium, Angelicae Gigantis Radix, Clematidis Radix, and Phellodendri Cortex. This study was designed to investigate its chondroprotective and anti-inflammatory effects to develop an anti-arthritic herb medicine. METHODS: ChondroT was validated using a convenient and accurate high-performance liquid chromatography-photodiode array (HPLC-PDA) detection method for simultaneous determination of its seven reference components. The concentrations of the seven marker constituents were in the range of 0.81-5.46 mg/g. The chondroprotective effects were evaluated based on SW1353 chondrocytes and matrix metalloproteinase 1 (MMP1) expression. In addition, the anti-inflammatory effects of ChondroT were studied by Western blotting of pro-inflammatory enzymes and by enzyme-linked immunosorbent assay (ELISA) of inflammatory mediators in lipopolysaccharides (LPS)-induced RAW264.7 cells. RESULTS: ChondroT enhanced the growth of SW1353 chondrocytes and also significantly inhibited IL-1ß-induced MMP-1 expression. However, ChondroT did not show any effects on the growth of HeLa and RAW264.7 cells. The expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) was induced by LPS in RAW264.7 cells, which was significantly decreased by pre-treatment with ChondroT. In addition, ChondroT reduced the activation of NF-kB and production of inflammatory mediators, such as IL-1ß, IL-6, PGE2, and nitric oxide (NO) in LPS-induced RAW264.7 cells. CONCLUSIONS: These results show that ChondroT exerted a chondroprotective effect and demonstrated multi-target mechanisms related to inflammation and arthritis. In addition, the suppressive effect was greater than that exhibited by GHJTY, suggesting that ChondroT, a new complex herbal medication, has therapeutic potential for the treatment of arthritis.

Evaluation of Anti-Inflammatory Potential of the New Ganghwaljetongyeum on Adjuvant-Induced Inflammatory Arthritis in Rats.

Ganghwaljetongyeum (GHJTY) has been used as a standard treatment for arthritis for approximately 15 years at the Korean Medicine Hospital of Dongshin University. GHJTY is composed of 18 medicinal herbs, of which five primary herbs were selected and named new Ganghwaljetongyeum (N-GHJTY). The purpose of the present study was to observe the effect of N-GHJTY on arthritis and to determine its mechanism of action. After confirming arthritis induction using complete Freund's adjuvant (CFA) in rats, N-GHJTY (62.5, 125, and 250 mg/kg/day) was administered once a day for 10 days. In order to determine pathological changes, edema of the paws and weight were measured before and for 10 days after N-GHJTY administration. Cytokine (TNF-α, IL-1ß, and IL-6) levels and histopathological lesions in the knee joint were also examined. Edema in the paw and knee joint of N-GHJTY-treated rats was significantly decreased at 6, 8, and 10 days after administration, compared to that in the CFA-control group, while weight consistently increased. Rats in N-GHJTY-treated groups also recovered from the CFA-induced pathological changes and showed a significant decline in cytokine levels. Taken together, our results showed that N-GHJTY administration was effective in inhibiting CFA-induced arthritis via anti-inflammatory effects while promoting cartilage recovery by controlling cytokine levels.

HerDing: herb recommendation system to treat diseases using genes and chemicals.

In recent years, herbs have been researched for new drug candidates because they have a long empirical history of treating diseases and are relatively free from side effects. Studies to scientifically prove the medical efficacy of herbs for target diseases often spend a considerable amount of time and effort in choosing candidate herbs and in performing experiments to measure changes of marker genes when treating herbs. A computational approach to recommend herbs for treating diseases might be helpful to promote efficiency in the early stage of such studies. Although several databases related to traditional Chinese medicine have been already developed, there is no specialized Web tool yet recommending herbs to treat diseases based on disease-related genes. Therefore, we developed a novel search engine, HerDing, focused on retrieving candidate herb-related information with user search terms (a list of genes, a disease name, a chemical name or an herb name). HerDing was built by integrating public databases and by applying a text-mining method. The HerDing website is free and open to all users, and there is no login requirement. Database URL: http://combio.gist.ac.kr/herding.

Tumor homing indocyanine green encapsulated micelles for near infrared and photoacoustic imaging of tumors.

Photoacoustic imaging (PAI) is an emerging analytical modality that is under intense preclinical development for the early diagnosis of various medical conditions, including cancer. However, the lack of specific tumor targeting by various contrast agents used in PAI obstructs its clinical applications. In this study, we developed indocyanine green (ICG)-encapsulated micelles specific for the CD 44 receptor and used in near infrared and photoacoustic imaging of tumors. ICG was hydrophobically modified prior to loading into hyaluronic acid (HA)-based micelles utilized for CD 44 based-targeting. We investigated the physicochemical characteristics of prepared HA only and ICG-encapsulated HA micelles (HA-ICG micelles). After intravenous injection of tumor-bearing mice, the bio-distribution and in vivo photoacoustic images of ICG-encapsulated HA micelles accumulating in tumors were also investigated. Our study further encourages the application of this HA-ICG-based nano-platform as a tumor-specific contrast agent for PAI.