Wound healing is promoted by Musa paradisiaca (banana) extract in diabetic rats.

Introduction: Impairments in wound healing commonly occur among patients with diabetes. Herbal medicines have a long history of usage in wound care management. Super green (SG) is a newly discovered natural product obtained from Musa paradisiaca. This study aimed to investigate the efficacy of the topical application of SG in healing surgical wounds in diabetic rats. Material and methods: Wistar rats received a one-time intraperitoneal injection of streptozotocin to induce type 1 diabetes. Full-thickness excisional skin wounds were created on the backs of the rats. The relevant groups were topically treated with the indicated concentrations of SG or vehicle dressing throughout the study duration. Histological analysis was performed and the mRNA levels of proinflammatory cytokines were measured to evaluate the improvement of wound closure. Results: The wound area ratio of the SG (1/6000 dilution)-treated group was greatly reduced compared to that of the vehicle-treated group. The histological analysis showed fewer inflammatory cells, accelerated re-epithelialization, and increased collagen deposition in SG 1/6000-treated wounds. The gene expression levels of tumor necrosis factor-α, interleukin-1ß, and interleukin-6 were decreased and the levels of type I and type III collagen were increased after SG treatment. Conclusions: These results show that the most therapeutically efficacious concentration of SG (1/6000 dilution) can enhance wound repair in diabetic rats. SG has the potential to be a new treatment strategy for diabetic wounds.

Evaluating cluster analysis techniques in ChatGPT versus R-language with visualizations of author collaborations and keyword cooccurrences on articles in the Journal of Medicine (Baltimore) 2023: Bibliometric analysis.

BACKGROUND: Analyses of author collaborations and keyword co-occurrences are frequently used in bibliographic research. However, no studies have introduced a straightforward yet effective approach, such as utilizing ChatGPT with Code Interpreter (ChatGPT_CI) or the R language, for creating cluster-oriented networks. This research aims to compare cluster analysis methods in ChatGPT_CI and R, visualize country-specific author collaborations, and then demonstrate the most effective approach. METHODS: The research focused on articles and review pieces from Medicine (Baltimore) published in 2023. By August 20, 2023, we had gathered metadata for 1976 articles using the Web of Science core collections. The efficiency and effectiveness of cluster displays between ChatGPT_CI and R were compared by evaluating their time consumption. The best method was then employed to present a series of visualizations of country-specific author collaborations, rooted in social network and cluster analyses. Visualization techniques incorporating network charts, chord diagrams, circle bar plots, circle packing plots, heat dendrograms, dendrograms, and word clouds were demonstrated. We further highlighted the research profiles of 2 prolific authors using timeline visuals. RESULTS: The research findings include that (1) the most active contributors were China, Nanjing Medical University (China), the Medical School Department, and Dr Chou from Taiwan when considering countries, institutions, departments, and individual authors, respectively; (2) the highest cited articles originated from Medicine (Baltimore) accounting for 4.53%: New England Journal of Medicine, PLOS ONE, LANCET, and The Journal of the American Medical Association, with respective contributions of 3.25%, 2.7%, 2.52%, and 1.54%; (3) visual cluster analysis in R proved to be more efficient and effective than ChatGPT_CI, reducing the time taken from 1 hour to just 3 minutes; (4) 7 cluster-focused networks were crafted using R on a custom platform; and (5) the research trajectories of 2 prominent authors (Dr Brin from the United States and Dr Chow from Taiwan) and articles themes in Medicine 2023 were depicted using timeline visuals. CONCLUSIONS: This research highlighted the efficient and effective methods for conducting cluster analyses of author collaborations using R. For future related studies, such as keyword co-occurrence analysis, R is recommended as a viable alternative for bibliographic research.

Visual impact beam plots: Analyzing research profiles and bibliometric metrics using the following-leading clustering algorithm (FLCA).

BACKGROUND: A new approach to showcasing author publications on a website involves using a visual representation instead of the conventional paper list. The creation of an impact beam plot (IBP) as a research profile for individuals is crucial, especially when incorporating collection edges that include self-cited articles through a rare cluster analysis technique not commonly found in the literature. This study presents the application of a unique method called the following-leading clustering algorithm (FLCA) to generate IBPs for 3 highly productive authors. METHODS: For the 3 highly productive authors, Sung-Ho Jang from South Korea, Chia-Hung Kao from Taiwan, and Chin-Hsiao Tseng from Taiwan, all their published articles indexed in the Web of Science Core Collection were downloaded. Sung-Ho Jang published 593 articles, Chia-Hung Kao published 732 articles, and Chin-Hsiao Tseng published 160 articles. To analyze and showcase their publications, the FLCA was utilized. This algorithm helped cluster their articles and identify representative publications for each author. To assess the effectiveness and validity of the FLCA algorithm, both network charts and heatmaps with dendrograms were employed. IBPs were then created and compared for each of the 3 authors, taking into consideration their h-index, x-index, and self-citation rate. This allowed for a comprehensive visual representation of their research impact and citation patterns. RESULTS: The results show that these authors' h-index, x-index, and self-citation rates were (37, 44.01, 1.66%), (42, 61.47, 0.23%), and (37, 40.3, 6.62%), respectively. A higher value in these metrics indicates a more remarkable research achievement. A higher self-citation rate with a lower cluster number indicates that manuscripts are more likely to have been self-drafted. Using the FLCA algorithm, IBPs were successfully generated for each author. CONCLUSION: The FLCA algorithm allows for the easy generation of visual IBPs based on authors' publication profiles. These IBPs incorporate 3 important bibliometric metrics: h-index, x-index, and self-citations. These metrics are highly recommended for use by researchers globally, particularly with the self-citation rate, as they offer valuable insights into the scholarly impact and citation patterns of individual researchers.

Free Myocutaneous Flap Assessment in a Rat Model: Verification of a Wireless Bioelectrical Impedance Assessment (BIA) System for Vascular Compromise Following Microsurgery.

BACKGROUND: Microvascular tissue transfer is a common reconstructive procedure. We designed a bioelectrical impedance assessment (BIA) system for quantitative analysis of tissue status. This study attempts to verify it through the animal model. METHODS: The flaps of the rat model were monitored by the BIA system. RESULTS: The BIA variation of the free flap in the rat after the vascular compromise was recorded. The non-vascular ligation limbs of the same rat served as a control group. The bio-impedance in the experimental group was larger than the control group. The bio-impedances of both the thigh/feet flaps in the experimental group were increased over time. In the thigh, the difference in bio-impedance from the control group was first detected at 10 kHz at the 3rd and last at 1 kHz at the 6th h, after vascular compromise. The same finding was observed in the feet. Compared with the control group, the bio-impedance ratio (1 kHz/20 kHz) of the experimental group decreased with time, while their variation tendencies in the thigh and feet were similar. CONCLUSIONS: The flap may be monitored by the BIA for vascular status.

Molecular mechanisms regarding potassium bromate­induced cardiac hypertrophy without apoptosis in H9c2 cells.

Cardiac hypertrophy is commonly involved in cardiac injury. Oxidative stress can induce cardiac hypertrophy with apoptosis. Potassium bromate (KBrO3) has been widely used as a food additive due to its oxidizing properties. In the present study, the rat­derived heart cell line H9c2 was used to investigate the effect of KBrO3 on cell size. KBrO3 increased cell size at concentrations <250 µM, in a dose­dependent manner. Additionally, KBrO3 also promoted the gene expression of two biomarkers of cardiac hypertrophy, brain/B­type natriuretic peptides (BNP) and ß­Myosin Heavy Chain (ß­MHC). However, apoptosis remained unobserved in these cells. Moreover, mediation of free radicals was investigated using a fluorescence assay, and it was observed that superoxide and reactive oxygen species (ROS) levels increased with KBrO3. Effects of KBrO3 were significantly reduced by tiron at concentrations sufficient to produce antioxidant­like action. Additionally, signals involved in cardiac hypertrophy such as calcineurin and nuclear factor of activated T­cells (NFAT) were also determined using western blot analysis. KBrO3 increased the protein levels of both these molecules which were decreased by tiron in a dose­dependent manner. Additionally, cyclosporine A attenuated the cardiac hypertrophy induced by KBrO3 in H9c2 cells at concentrations effective to inhibit calcineurin, in addition to reducing mRNA levels of BNP or ß­MHC. Finally, apoptosis was also identified in H9c2 cells incubated with KBrO3 at concentrations >300 µM. Collectively, these results provided a novel perspective that KBrO3 induces cardiac hypertrophy without apoptosis at a low dose through the generation of ROS, activating the calcineurin/NFAT signaling pathway in H9c2 cells. Therefore, at a dose <250 µM, KBrO3 can be applied as an inducer of cardiac hypertrophy without apoptosis in H9c2 cells. KBrO3 can also be developed as a tool to induce cardiac hypertrophy in animals.

GW0742 activates peroxisome proliferator-activated receptor δ to reduce free radicals and alleviate cardiac hypertrophy induced by hyperglycemia in cultured H9c2 cells.

Peroxisome proliferator-activated receptor δ (PPARδ), the predominant PPAR subtype in the heart, is known to regulate cardiac function. PPARδ activation may inhibit cardiac hypertrophy in H9c2 cells while the potential mechanism has not been elucidated. Then, H9c2 cells incubated with high glucose to induce hypertrophy were used to investigate using GW0742 to activate PPARδ. The fluorescence assays were applied to determine the changes in cell size, cellular calcium levels, and free radicals. Western blot analyses for hypertrophic signals and assays of messenger RNA (mRNA) levels for hypertrophic biomarkers were performed. In H9c2 cells, GW0742 inhibited cardiac hypertrophy. In addition, increases in cellular calcium and hypertrophic signals, including calcineurin and nuclear factor of activated T-cells, were reduced by GW0742. This reduction was parallel to the decrease in the mRNA levels of biomarkers, such as brain/B-type natriuretic peptides and ß-myosin heavy chain. These effects of GW0742 were dose-dependently inhibited by GSK0660 indicating an activation of PPARδ by GW0742 to alleviate cardiac hypertrophy. Moreover, free radicals produced by hyperglycemia were also markedly inhibited by GW0742 and were later reversed by GSK0660. GW0742 promoted the expression of thioredoxin, an antioxidant enzyme. Direct inhibition of reactive oxygen species by GW0742 was also identified in the oxidant potassium bromate stimulated H9c2 cells. Taken together, these findings suggest that PPARδ agonists can inhibit free radicals, resulting in lower cellular calcium for reduction of hypertrophic signaling to alleviate cardiac hypertrophy in H9c2 cells. Therefore, PPARδ activation can be used to develop agent(s) for treating cardiac hypertrophy.

Phosphorylation of signal transducer and activator of transcription 3 induced by hyperglycemia is different with that induced by lipopolysaccharide or erythropoietin via receptor­coupled signaling in cardiac cells.

The signal transducer and activator of transcription 3 (STAT3) is known to be involved in hypertrophy and fibrosis in cardiac dysfunction. The activation of STAT3 via the phosphorylation of STAT3 is required for the production of functional activity. It has been established that lipopolysaccharide (LPS)­induced phosphorylation of STAT3 in cardiomyocytes primarily occurs through a direct receptor­mediated action. This effect is demonstrated to be produced rapidly. STAT3 in cardiac fibrosis of diabetes is induced by high glucose through promotion of the STAT3­associated signaling pathway. However, the time schedule for STAT3 activation between LPS and high glucose appears to be different. Therefore, the difference in STAT3 activation between LPS and hyperglycemia in cardiomyocytes requires elucidation. The present study investigated the phosphorylation of STAT3 induced by LPS and hyperglycemia in the rat cardiac cell line H9c2. Additionally, phosphorylation of STAT3 induced by erythropoietin (EPO) via receptor activation was compared. Then, the downstream signals for fibrosis, including the connective tissue growth factor (CTGF) and matrix metalloproteinase (MMP)­9, were determined using western blotting, while the mRNA levels were quantified. LPS induced a rapid elevation of STAT3 phosphorylation in H9c2 cells within 30 min, similar to that produced by EPO. However, LPS or EPO failed to modify the mRNA level of STAT3, and/or the downstream signals for fibrosis. High glucose increased STAT3 phosphorylation to be stable after a long period of incubation. Glucose incubation for 24 h may augment the STAT3 expression in a dose­dependent manner. Consequently, fibrosis­associated signals, including CTGF and MMP­9 protein, were raised in parallel. In the presence of tiron, an antioxidant, these changes by hyperglycemia were markedly reduced, demonstrating the mediation of oxidative stress. Therefore, LPS­ or EPO­induced STAT3 phosphorylation is different compared with that caused by high glucose in H9c2 cells. Sustained activation of STAT3 by hyperglycemia may promote the expression of fibrosis­associated signals, including CTGF and MMP­9, in H9c2 cells. Therefore, regarding the cardiac dysfunctions associated with diabetes and/or hyperglycemia, the identification of nuclear STAT3 may be more reliable compared with the assay of phosphorylated STAT3 in cardiac cells.

Loperamide-induced Cardiac Depression Is Enhanced by Hyperglycemia: Evidence Relevant to Loperamide Abuse.

BACKGROUND AND AIMS: Cardiac dysryhthmias and death are reported after loperamide abuse. The mechanism of death is not clear and cardiac depression may play a role in this mechanism. Loperamide is widely used as an agonist of the µ-opioid receptor (MOR) in clinical practice. In skeletal muscle, an increase in MOR in response to hyperglycemia is largely attributable to higher expression of the transducer and activator of transcription 3 (STAT3), which binds to the promoter of the MOR genes. Therefore, we investigated the changes in cardiac MOR caused by hyperglycemia both in vivo and in vitro. METHODS: Streptozotocin-induced type 1-like diabetic rats (STZ rats) were used to estimate cardiac performance and changes in cardiac MOR under the influence of loperamide. STAT3 was measured in cultured cardiomyocytes under high glucose (HG) to mimic the in vivo changes. RESULTS: Loperamide-induced reduction of cardiac performance was more marked in STZ rats than in normal rats. The increased MOR in the hearts of STZ rats was reversed by the reduction of hyperglycemia. Higher MOR expression paralleled the increase in STAT3 in cardiomyocytes under HG and was reversed by siRNA of STAT3. Stattic at a dose sufficient to inhibit STAT3 reduced MOR both in vivo and in vitro. CONCLUSION: Cardiac depression induced by loperamide is enhanced by hyperglycemia due to higher MOR expression, which is associated with higher expression of STAT3 in the heart. These results suggest that loperamide abuse is particularly dangerous for individuals with hyperglycemia.

Rhodiola-water extract induces ß-endorphin secretion to lower blood pressure in spontaneously hypertensive rats.

Rhodiola rosea (Rhodiola) is grown at high altitudes and northern latitudes. It is mainly used clinically as an adaptogen, but antihypertensive effects have been reported for the extract. These have not been well investigated, so in the present study, we evaluated the effect of Rhodiola-water extract on blood pressure in spontaneously hypertensive rats (SHRs) and investigated the potential mechanism(s) for this action. In conscious male SHRs, systolic blood pressure (SBP) and heart rate were recorded using the tail-cuff method. Plasma ß-endorphin was measured by enzyme-linked immunosorbent assay. Rhodiola-water extract decreased SBP in SHRs in a dose-dependent manner, and this action was more significant than that in normal group named Wistar-Kyoto (WKY) rats. This reduction of SBP in SHRs was inhibited by pretreatment with the selective opioid µ-receptor antagonist, cyprodime, but not by naloxonazine, an antagonist specific to opioid µ1-receptor. Also, the SBP-lowering action of Rhodiola-water extract was attenuated in adrenalectomized SHRs. Moreover, Rhodiola-water extract dose-dependently increased ß-endorphin release in SHRs, and the elevation of ß-endorphin in SHRs was higher than that in WKY. Thus, we suggest that Rhodiola-water extract can induce release of ß-endorphin to lower SBP in SHRs.

Increase of myocardial performance by Rhodiola-ethanol extract in diabetic rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Rhodiola rosea (also known as golden root or roseroot) is a perennial plant of the Crassulaceae family that grows in the Arctic and in the mountainous regions of Europe, Asia, and North America. The rhizome and roots of this plant have been long used as traditional medicine in Eastern Europe and Asia for enhancing physical and mental performance. AIM OF THE STUDY: The present study is designed to investigate the cardiac action of Rhodiola-ethanol extract in streptozotocin-induced diabetic rats (STZ-diabetic rats) with heart failure. MATERIALS AND METHODS: Diabetes was induced in Wistar rats by injection of streptozotocin. We measured the changes of body weight, water intake, and food intake in three groups of age-matched rats; the normal control received vehicle, STZ-diabetic rat received Rhodiola-ethanol extract or vehicle. Cardiac output, heart rate, blood pressure, and hemodynamic dP/dt in addition to plasma insulin and glucose level were also determined. The mRNA and protein levels of PPARδ were measured using real-time PCR and Western blotting, respectively. RESULTS: Food intake, water intake and blood glucose were raised in STZ-diabetic rats showing lower body weight and plasma insulin, as compared with the control. Also, cardiac output, heart rate, blood pressure and hemodynamic dP/dt were markedly reduced in STZ-diabetic rats indicating the heart failure physiologically. After a 21-day treatment with Rhodiola-ethanol extract, cardiac output was raised in STZ-rats while the diabetic parameters were not modified. The PPARδ expression of both mRNA and protein was markedly elevated in the heart of STZ-rats receiving treatment with Rhodiola-ethanol extract. Also, the increased phosphorylation level of cardiac troponin-I was restored by this treatment with Rhodiola-ethanol extract. Otherwise, increase of cardiac output by Rhodiola-ethanol extract was blocked by antagonist of PPARδ in STZ-diabetic rats. CONCLUSIONS: Our results suggest that ethanol extract of Rhodiola has an ability to increase the cardiac output in STZ-diabetic rats showing heart failure. Also, increase of PPAR-δ is responsible for this action of Rodiola-ethanol extract.

Increase of ATP-sensitive potassium (K(ATP)) channels in the heart of type-1 diabetic rats.

BACKGROUND: An impairment of cardiovascular function in streptozotocin (STZ)-diabetic rats has been mentioned within 5 days-to-3 months of induction. ATP-sensitive potassium (K(ATP)) channels are expressed on cardiac sarcolemmal membranes. It is highly responsive to metabolic fluctuations and can have effects on cardiac contractility. The present study attempted to clarify the changes of cardiac K(ATP) channels in diabetic disorders. METHODS: Streptozotocin-induced diabetic rats and neonatal rat cardiomyocytes treated with a high concentration of glucose (a D-glucose concentration of 30 mM was used and cells were cultured for 24 hr) were used to examine the effect of hyperglycemia on cardiac function and the expression of K(ATP) channels. K(ATP) channels expression was found to be linked to cardiac tonic dysfunction, and we evaluated the expression levels of K(ATP) channels by Western blot and Northern blot analysis. RESULTS: The result shows diazoxide produced a marked reduction of heart rate in control group. Furthermore, the methods of Northern blotting and Western blotting were employed to identify the gene expression of K(ATP) channel. Two subunits of cardiac K(ATP) channel (SUR2A and kir 6.2) were purchased as indicators and showed significantly decreased in both diabetic rats and high glucose treated rat cardiac myocytes. Correction of hyperglycemia by insulin or phlorizin restored the gene expression of cardiac K(ATP) in these diabetic rats. CONCLUSIONS: Both mRNA and protein expression of cardiac K(ATP) channels are decreased in diabetic rats induced by STZ for 8 weeks. This phenomenon leads to result in desensitization of some K(ATP) channel drugs.