Photo-triggered caffeic acid delivery via psyllium polysaccharide- gellan gum-based injectable bionanogel for epidermoid carcinoma treatment.

Here, the simultaneous effect of chemo- and photothermal therapy against epidermoid carcinoma (EC) was investigated. A novel hydrogel, termed bionanogel (BNG), was designed using psyllium mucilage polysaccharide and bacterial gellan gum, incorporated with nanocomplex carrying caffeic acid (CA) and IR-820, and further characterized. The dual effect of BNG and 808 nm laser (BNG + L) on EC was investigated. Staining and scratch assays were performed to analyze their therapeutic effect on EC. In vivo evaluations of BNG + L in xenograft models were performed. Rapid transition, limited swelling, degradability and high tensile strength indicated BNG stability and sustained drug release. Irradiation with 808 nm laser light at 1.25 W /cm2 for 4 min resulted in a temperature increase of 53 °C and facilitated cell ablation. The in vitro studies showed that BNG + L suppressed cancer progression via a late apoptotic effect. The in vivo study showed that the slow release of CA from BNG + L significantly attenuated EC with low mitotic index and downregulation of proteins involved in cancer proliferation such as EGFR, AKT, PI3K, ERK, mTOR and HIF-1α. Thus, BNG could be a novel medium for targeted and controlled drug delivery for the treatment of epidermoid cancer when triggered by NIR light.

Quantitative association between gene expression and blood cell production of individual hematopoietic stem cells in mice.

Individual hematopoietic stem cells (HSCs) produce different amounts of blood cells upon transplantation. Taking advantage of the intercellular variation, we developed an experimental and bioinformatic approach to evaluating the quantitative association between gene expression and blood cell production across individual HSCs. We found that most genes associated with blood production exhibit the association only at some levels of blood production. By mapping gene expression with blood production, we identified four distinct patterns of their quantitative association. Some genes consistently correlate with blood production over a range of levels or across all levels, and these genes are found to regulate lymphoid but not myeloid production. Other genes exhibit one or more clear peaks of association. Genes with overlapping peaks are found to be coexpressed in other tissues and share similar molecular functions and regulatory motifs. By dissecting intercellular variations, our findings revealed four quantitative association patterns that reflect distinct dose-response molecular mechanisms modulating the blood cell production of HSCs.

Effect of dual-optical pulses with temporal energy distribution on laser ablation performance in in vivo zebrafish model.

A Q-switched laser system has been used in a single-pulse mode for skin melasma treatments because of instant heat deposition in the target. Despite the efficient ablation of the melanophores in the skin, the single, high-fluence pulse often causes undesirable damage to the surrounding tissue, leading to high recurrence rates. This study aims to investigate the feasibility of dual-optical pulses with a temporal energy distribution on the melasma treatment in in vivo zebrafish models in comparison to that of the single optical pulse. Based on the optical detection, the dual-optical pulses had a temporal energy distribution ratio of 4:1 and an interval of 61 µs between the two consecutive pulses. According to the histological analysis, the dual pulses removed melanophores and induced a few apoptotic nuclei with minimal recurrence. This study demonstrated that the feasibility of dual-optical pulses (energy ratio = 4:1) could enhance the laser ablation performance in vivo.

Anti-Cancer Effect of Chlorophyllin-Assisted Photodynamic Therapy to Induce Apoptosis through Oxidative Stress on Human Cervical Cancer.

Photodynamic therapy is an alternative approach to treating tumors that utilizes photochemical reactions between a photosensitizer and laser irradiation for the generation of reactive oxygen species. Currently, natural photosensitive compounds are being promised to replace synthetic photosensitizers used in photodynamic therapy because of their low toxicity, lesser side effects, and high solubility in water. Therefore, the present study investigated the anti-cancer efficacy of chlorophyllin-assisted photodynamic therapy on human cervical cancer by inducing apoptotic response through oxidative stress. The chlorophyllin-assisted photodynamic therapy significantly induced cytotoxicity, and the optimal conditions were determined based on the results, including laser irradiation time, laser power density, and chlorophyllin concentration. In addition, reactive oxygen species generation and Annexin V expression level were detected on the photodynamic reaction-treated HeLa cells under the optimized conditions to evaluate apoptosis using a fluorescence microscope. In the Western blotting analysis, the photodynamic therapy group showed the increased protein expression level of the cleaved caspase 8, caspase 9, Bax, and cytochrome C, and the suppressed protein expression level of Bcl-2, pro-caspase 8, and pro-caspase 9. Moreover, the proposed photodynamic therapy downregulated the phosphorylation of AKT1 in the HeLa cells. Therefore, our results suggest that the chlorophyllin-assisted photodynamic therapy has potential as an antitumor therapy for cervical cancer.

RNA splicing factor Rbm25 underlies heterogeneous preleukemic clonal expansion in mice.

Clonal expansion sets the stage for cancer genesis by allowing for the accumulation of molecular alterations. Although genetic mutations such as Tet2 that induce clonal expansion and malignancy have been identified, these mutations are also frequently found in healthy individuals. Here, we tracked preleukemic clonal expansion using genetic barcoding in an inducible Tet2 knockout mouse model and found that only a small fraction of hematopoietic stem cells (HSCs) expanded excessively upon Tet2 knockout. These overexpanded HSCs expressed significantly lower levels of genes associated with leukemia and RNA splicing than nonoverexpanded Tet2 knockout HSCs. Knocking down Rbm25, an identified RNA splicing factor, accelerated the expansion of Tet2-knockout hematopoietic cells in vitro and in vivo. Our data suggest that mutations of an epigenetic factor Tet2 induce variability in the expression of an RNA splicing factor Rbm25, which subsequently drives heterogeneous preleukemic clonal expansion. This heterogeneous clonal expansion could contribute to the variable disease risks across individuals.

Photomodulative effects of low-level laser therapy on tracheal fenestration developed in in vivo model.

The effect of low-level laser therapy (LLLT) on variable mucosal lesions in the upper aerodigestive tract has been reported. However, the effect of LLLT on tracheostomy sites or tracheal fenestration is rarely reported. In this study, we evaluate the effect of LLLT performed using 635 nm laser light based on a cylindrical diffuser and an animal model with tracheal fenestration. An animal model of tracheal fenestration is developed by suturing the trachea to the skin after performing a vertical tracheostomy from the second to the fifth tracheal ring of Wistar rats (male, body weight 200-250 g). LLLT (spot size: 2 cm2) is conducted once daily for five days using a handheld cylindrical device. Twenty-four rats are randomly assigned to a no-therapy or LLLT group with an energy density of 20 J/cm2. Histological analysis is performed at 7 and 14 days after tracheal fenestration. Irradiation at the tracheal fenestration site with an energy density of 20 J/cm2 improves the wound healing, as shown at 2 weeks after tracheostomy. Histological analysis shows significantly decreased acute inflammation and granulation tissue, as well as better cartilage regeneration and less tracheal wall thickening. Therefore, LLLT demonstrates therapeutic potential for preventing tracheal stenosis and granuloma after tracheostomy.

Cellulose nanofibrils reinforced chitosan-gelatin based hydrogel loaded with nanoemulsion of oregano essential oil for diabetic wound healing assisted by low level laser therapy.

Diabetic foot ulcers are imperfections in the process of wound healing due to hyperglycemic conditions. Here, a nanoemulgel fabricated with oregano essential oil nanoemulsion, assisted by low-level laser therapy, was investigated for its efficacy in diabetic wound healing. A hydrogel- based healing patch, fabricated using biological polymers namely chitosan and gelatin and, polyvinyl pyrollidone. The hydrogel was reinforced with cellulose nanofibrils for enhanced stability and barrier properties. Nanoemulsion of oregano essential oil, with an average particle size of 293.7 ± 8.3 nm, was prepared via homogenization with chitosan as the coating agent. Nanoemulsion impregnated hydrogel, termed as the nanoemulgel, was assessed for its physio-mechanical properties and healing efficiency. The strong linkages in nanoemulgel demonstrated its large swelling capacity, high mechanical strength, and maximum thermal stability. The optimized conditions for low-level laser therapy using 808 nm were 1 W. cm-2 and 5 min. The optimized drug concentration of 128 µg. mL-1 exhibited viability of NIH/3 T3 fibroblasts as 75.5 ± 1.2 % after 24 h. Cell migration assay demonstrated that dual therapy facilitated wound healing, with a maximum closure rate of 100 % at 48 h. In vivo results revealed the rapid healing effects of the dual therapy in diabetic rat models with foot ulcers: a maximum healing rate of 97.5 %, minimum scar formation, increased granulation, enhanced reepithelialization, and a drastic decrease in inflammation and neutrophil infiltration within the treatment period compared to monotherapy and control. In summary, the combinatorial therapy of nanoemulgel and low-level laser therapy is a promising regimen for managing diabetic foot ulcers with a rapid healing effect.

Combinatorial prophylactic effect of phlorotannins with photobiomodulation against tracheal stenosis.

Several conventional treatments are used to manage tracheal stenosis after intubation and surgical procedures; however, patients are at risk of restenosis because of the absence of effective preventative therapy. In this study, we evaluate the biomodulatory effect of PT-combined blue light (BL) PBM in tracheostomal stenosis-induced animal models. The PT-combined BL group showed a significant decrease in the fibrotic protein synthesis by downregulating the release of stenosis-triggering fibrotic signals, without cytotoxicity or thermal damage. Moreover, the combined treatment ameliorated excessive granulation and collagen formation, and consequently preserved the opening of the tracheostoma ten days after fenestration. The current study demonstrated the biomodulatory effect of PT-combined BL on human tracheal fibroblasts and tracheal fenestration rodent models. Hence, PT-combined BL has the potential to be an effective preventative treatment for tracheal stenosis but also as an alternative option for fibrotic disorders.

Comparison of the triglyceride glucose (TyG) index, triglyceride to high-density lipoprotein cholesterol (TG/HDL-C) ratio, and metabolic score for insulin resistance (METS-IR) associated with periodontitis in Korean adults.

Background: Periodontitis is one of the most common diseases associated with the oral cavity. Previous studies have suggested that there is an association between periodontitis and metabolic dysfunction. Recently, the triglyceride glucose (TyG) index, high-density lipoprotein cholesterol (TG/HDL-C) ratio, and metabolic score for insulin resistance (METS-IR) index have been identified as useful markers for assessing insulin resistance. Objective: This study aimed to evaluate the relationship between periodontitis and non-insulin-based insulin resistance (IR) indices and compare the predictive values of these indices in the Korean population. Design: This is a cross-sectional study. Methods: A total of 13,584 participants were included in the 2013-2015 Korean National Health and Nutrition Examination Survey data. A community periodontal index score⩾3 was used to define periodontitis. Participants were divided into quartiles according to each index. Odds ratios (ORs) and 95% confidence intervals (CIs) for the prevalence of periodontitis and the TyG index, TG/HDL-C ratio, and METS-IR index quartiles were calculated using multiple logistic regression analysis. We estimated the areas under the receiver operating characteristic curves (AUCs) of the indices to compare the predictive values of the three indices. Results: Compared with quartile 1, the fourth quartile ORs (95% CIs) for periodontitis were 1.23 (1.01-1.49) for the TyG index, 1.23 (1.02-1.48) for the TG/HDL-C ratio, and 1.53 (1.25-1.88) for the METS-IR index after adjustment for confounders. The AUC (95% CIs) was 0.608 (0.598-0.618) for the TyG index, 0.600 (0.590-0.610) for the TG/HDL-C ratio, and 0.617 (0.608-0.627) for the METS-IR index to identify periodontitis. The predictive power of METS-IR was significantly higher than that of the TyG index and TG/HDL-C. Conclusion: Higher TG/HDL-C ratio, TyG, and METS-IR indices are associated with a higher prevalence of periodontitis. The METS-IR index is a more powerful predictor of periodontitis prevalence than the TyG index and TG/HDL-C ratio.

Evaluation of blood perfusion using laser doppler flowmetry during endoscopic lumbar sympathectomy in patients with plantar hyperhidrosis: a retrospective observational study.

The purpose of this study is to evaluate whether monitoring the changes of skin blood flow may be effective in assessing blood perfusion during endoscopic lumbar sympathectomy (ELS) in patients with plantar hyperhidrosis. In this study, a total of 30 patients who underwent surgical treatment for plantar hyperhidrosis at the Department of Thoracic and Cardiovascular Surgery in Yonsei University Gangnam Severance Hospital, Seoul, Korea, between July 2020 and December 2020, were retrospectively analyzed. Sympathetic denervation was performed on the third lumbar ganglion, and intraoperative laser doppler flowmetry (LDF) was used to detect the lumbar sympathetic chain accurately. We observed an abrupt increase of peripheral blood flow after sympathetic denervation, and the median percent changes of perfusion unit were 173.27 (inter-quartile range, IQR 195.48) and 392.98 (IQR 597.27) for the left and right sympathectomies, respectively. This study demonstrated the efficacy of monitoring skin blood flow via LDF during ELS. This result suggests that exact detection of blood flow using LDF is essential for improving the accuracy of ELS by checking the perfusion site on the sole in patients with plantar hyperhidrosis.

Automated Structural Analysis and Quantitative Characterization of Scar Tissue Using Machine Learning.

An analysis of scar tissue is necessary to understand the pathological tissue conditions during or after the wound healing process. Hematoxylin and eosin (HE) staining has conventionally been applied to understand the morphology of scar tissue. However, the scar lesions cannot be analyzed from a whole slide image. The current study aimed to develop a method for the rapid and automatic characterization of scar lesions in HE-stained scar tissues using a supervised and unsupervised learning algorithm. The supervised learning used a Mask region-based convolutional neural network (RCNN) to train a pattern from a data representation using MMDetection tools. The K-means algorithm characterized the HE-stained tissue and extracted the main features, such as the collagen density and directional variance of the collagen. The Mask RCNN model effectively predicted scar images using various backbone networks (e.g., ResNet50, ResNet101, ResNeSt50, and ResNeSt101) with high accuracy. The K-means clustering method successfully characterized the HE-stained tissue by separating the main features in terms of the collagen fiber and dermal mature components, namely, the glands, hair follicles, and nuclei. A quantitative analysis of the scar tissue in terms of the collagen density and directional variance of the collagen confirmed 50% differences between the normal and scar tissues. The proposed methods were utilized to characterize the pathological features of scar tissue for an objective histological analysis. The trained model is time-efficient when used for detection in place of a manual analysis. Machine learning-assisted analysis is expected to aid in understanding scar conditions, and to help establish an optimal treatment plan.

Deciphering intratumoral heterogeneity using integrated clonal tracking and single-cell transcriptome analyses.

Cellular heterogeneity is a major cause of treatment resistance in cancer. Despite recent advances in single-cell genomic and transcriptomic sequencing, it remains difficult to relate measured molecular profiles to the cellular activities underlying cancer. Here, we present an integrated experimental system that connects single cell gene expression to heterogeneous cancer cell growth, metastasis, and treatment response. Our system integrates single cell transcriptome profiling with DNA barcode based clonal tracking in patient-derived xenograft models. We show that leukemia cells exhibiting unique gene expression respond to different chemotherapies in distinct but consistent manners across multiple mice. In addition, we uncover a form of leukemia expansion that is spatially confined to the bone marrow of single anatomical sites and driven by cells with distinct gene expression. Our integrated experimental system can interrogate the molecular and cellular basis of the intratumoral heterogeneity underlying disease progression and treatment resistance.

Comparison of EQ-5D-3L and metabolic components between patients with hyperhidrosis and the general population: a propensity score matching analysis.

PURPOSE: It is important to understand the characteristics of patients with hyperhidrosis, which are different from the general population, for treating hyperhidrosis. Sympathetic overactivity, which might play an important role in hyperhidrosis, can contribute to metabolic diseases and the decreased quality of life (QoL). We compared the metabolic components and health-related QoL between patients with hyperhidrosis and the general population. METHODS: We conducted a case-control study and compared the characteristics of the patients (N = 196) with hyperhidrosis and propensity score-matched controls (N = 196) selected from the Korean National Health and Nutrition Examination Survey. Metabolic components and EQ-5D-3L (EQ-5D) index were compared using a two-way mixed analysis of covariance after adjusting for confounders. RESULTS: Patients with hyperhidrosis had significantly higher waist circumference (estimated mean values ± SD for patients and the control group, 85.5 ± 10.8 cm vs 81.3 ± 10.3 cm, p < 0.001), blood pressure (SBP, 121.1 ± 16.9 vs 111.7 ± 10.3, p < 0.001 AND DBP, 77.5 ± 12.8 vs 73.6 ± 8.6, p < 0.001, respectively), fasting glucose (97.1 ± 11.3 vs 91.5 ± 9.2, p < 0.001), and the number of components of metabolic syndrome (1.4 ± 1.3 vs 1.0 ± 1.2, p = 0.002), and significantly lower estimated glomerular filtration rate (144.3 ± 53.2 vs 158.3 ± 55.7, p = 0.002) and EQ-5D values (estimated mean values (standard error) for patients and the control group, 0.92 (0.01) vs 0.97 (0.01), p < 0.001) compared to the control group after adjustment. CONCLUSION: The patients with hyperhidrosis had more central obesity and unfavorable metabolic parameters and a lower EQ-5D index compared with the general population, emphasizing clinical importance of hyperhidrosis to be cured in aspect of metabolic components as well as patients' QOL.

Sex Differences in the Relationship Between High-Risk Drinking and the Triglyceride-Glucose (TyG) Index: An Analysis Using 2013 and 2015 Korean National Health and Nutrition Examination Survey Data.

AIMS: The alcohol use disorders identification test (AUDIT) was developed to evaluate excessive drinking in primary care. The triglyceride (TG) glucose (TyG) index is a novel marker used for assessing insulin resistance. We sought to document relationships between high-risk drinking according to AUDIT and the TyG index and to evaluate whether the TyG index is more correlated with high-risk drinking than TG or fasting plasma glucose (FPG). METHODS: We analyzed data for 7014 participants in the 2013 and 2015 Korea National Health and Nutrition Examination Surveys. Excessive drinking risk groups were categorized according to AUDIT scores (low-risk, 0-7 in men and 0-6 in women; moderate-risk, 8-14 in men and 7-12 in women; and high-risk, ≥15 in men and ≥13 in women). RESULTS: In men, compared with low-risk individuals, the odds ratios (95% confidence intervals) for higher TyG index values were 1.84 (1.16-2.93) in the moderate- and 2.82 (1.86-4.30) in the high-risk groups. The correlation coefficient for the TyG index and AUDIT score was significantly higher than those for TG and FPG. No significant associations were noted in women. CONCLUSION: High-risk drinking is significantly associated with higher TyG index values in men only. The TyG index can be a novel marker for assessing high-risk drinking in men.

U-Shaped Association between Sleep Duration, C-Reactive Protein, and Uric Acid in Korean Women.

Serum high-sensitivity C-reactive protein (hsCRP) and serum uric acid (SUA) are biomarkers that predict chronic inflammation and cardiovascular dysfunction. Therefore, we aimed to investigate the association between sleep duration, hsCRP, and SUA in Korean women. Cross-sectional data from the Seventh Korea National Health and Nutrition Examination Survey was analyzed. The odds ratio (OR) and 95% confidence intervals (CIs) for an association between higher hsCRP (>2.0 mg/L) or higher SUA (>5.6 mg/dL) and sleep duration were calculated using multiple logistic regression analyses after adjusting for potential confounders. In total, 6151 women were included in the analysis. There was a U-shaped relationship between continuous sleep duration, hsCRP, and SUA. Compared to those who slept for 7-8 h, the ORs (95% CIs) for higher hsCRP were 1.43 (0.95-2.16) in short sleepers and 1.64 (1.09-2.48) in long sleepers after adjusting for confounders. Compared with those who slept for 7-8 h, the ORs (95% CIs) for higher SUA were 1.54 (1.04-2.26) in short sleepers and 1.94 (1.27-2.96) in long sleepers after adjusting for confounders. We found a U-shaped association between sleep duration, hsCRP, and SUA in Korean women. 7-8 h sleep was associated with lower level of hsCRP and SUA in Korean women.

In vitro anti-tumor effect of low-power laser irradiation (LPLI) on gastroenterological carcinoma cells.

Primary hepatocellular carcinoma (HCC) and colon carcinoma are two of the most common clinical malignancies along with high morbidity and mortality. As low-power laser irradiation (LPLI) can induce cytotoxicity or cell apoptosis on several types of hyperplasia, LPLI may be a potential alternative treatment for gastroenterological cancers. The current in vitro study focused on LPLI-induced apoptosis and mechanism after 532-nm laser irradiation on two different carcinoma cells. Squamous cell carcinoma (VX2) and murine colon carcinoma (CT26) cells were cultured to test the feasibility of LPLI. The applied fluence varied from 0 to 600 J/cm2. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide analysis, fluorescence imaging, wound healing assay, and cell apoptosis tests were performed 24 h post-irradiation to monitor cellular responses. The current results demonstrated a dose-dependent stimulatory effect of LPLI on the cell viability, migration, and apoptosis of VX2 and CT26 cells. The therapeutic fluence of 600 J/cm2 induced statistically significant inhibition in cell viability. Both the wound healing assay and the cell apoptosis tests confirmed that LPLI with high fluences could inhibit cell migration as well as induce cell apoptosis. The current findings demonstrate that LPLI might be a potential treatment for the carcinoma cells. Further studies will be performed to evaluate the feasibility of LPLI in in vivo tumor models.

Combined treatment of low-level laser therapy and phloroglucinol for inhibition of fibrosis.

BACKGROUND AND OBJECTIVES: Fibrosis is a highly prevalent disease, which is responsible for 45% of deaths through pathological effects in developed countries. Previous studies have reported that low-level laser therapy (LLLT) can modulate fibrotic activity, but significant enhancement of therapeutic efficacy is still required for clinical translation. The aim of this study is to evaluate the feasible effect of LLLT combined with phloroglucinol (PHL) on the inhibition of fibrosis in vitro. STUDY DESIGN/MATERIALS AND METHODS: NIH/3T3 murine embryonic fibroblasts cells were cultured and transforming growth factor-ß1 (TGF-ß1) was treated for transition of fibroblasts. After TGF-ß1 treatment, LLLT and PHL were used, respectively, and in combination to suppress fibrosis. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and BrdU assays were performed to estimate the cell viability and proliferation. To evaluate the expression of fibrotic markers, we used confocal immunofluorescence and western blot. RESULTS: When compared with respectively treated groups, the group with the combined treatment of LLLT and PHL significantly reduced cell viability and proliferation. Immunofluorescence staining showed that the combined group minimized more α-smooth muscle actin (α-SMA) and type I collagen than the other groups. Western blot analysis showed that the combined treatment had significant decreases in α-SMA, TGF-ß1, and type I collagen. CONCLUSIONS: PHL-assisted LLLT may be an effective treatment to inhibit fibrosis due to its additive effects. The combined treatment has a potential to be an alternative treatment for fibrosis. Lasers Surg. Med. © 2019 Wiley Periodicals, Inc.