Pharmacological Tests Research Articles

Combination of Resminostat with Ruxolitinib Exerts Antitumor Effects in the Chick Embryo Chorioallantoic Membrane Model for Cutaneous T Cell Lymphoma.

Simple SummaryThe combination of Resminostat (HDACi) and Ruxolitinib (JAKi) exerted cytotoxic effects and inhibited proliferation of CTCL cell lines (MyLa, SeAx) in vitro. The aim of the present study was to validate their antitumor effects in vivo using the chick embryo chorioallantoic membrane (CAM) model, which allows quick and efficient monitoring of tumor growth, migration, invasion, and metastatic potential. The drug combination exhibited a significant inhibition of primary tumor size, and inhibited intravasation and extravasation of tumor cells to the liver and lung. It also exerted an inhibitory effect in the migration and invasion of tumor cells and significantly reduced key signaling pathway activation. Our data demonstrate that the CAM assay could be employed as a preclinical in vivo model in CTCL for pharmacological testing, and that the combination of Resminostat and Ruxolitinib exerts significant antitumor effects in CTCL progression that need to be further evaluated in a clinical setting.The combination of Resminostat (HDACi) and Ruxolitinib (JAKi) exerted cytotoxic effects and inhibited proliferation of CTCL cell lines (MyLa, SeAx) in previously published work. A xenograft tumor formation was produced by implanting the MyLa or SeAx cells on top of the chick embryo chorioallantoic membrane (CAM). The CAM assay protocol was developed to monitor the metastatic properties of CTCL cells and the effects of Resminostat and/or Ruxolitinib in vivo. In the spontaneous CAM assays, Resminostat and Ruxolitinib treatment inhibited the cell proliferation (p < 0.001) of MyLa and SeAx, and induced cell apoptosis (p < 0.005, p < 0.001, respectively). Although monotherapies reduced the size of primary tumors in the metastasis CAM assay, the drug combination exhibited a significant inhibition of primary tumor size (p < 0.0001). Furthermore, the combined treatment inhibited the intravasation of MyLa (p < 0.005) and SeAx cells (p < 0.0001) in the organs, as well as their extravasation to the liver (p < 0.0001) and lung (p < 0.0001). The drug combination also exerted a stronger inhibitory effect in migration (p < 0.0001) rather in invasion (p < 0.005) of both MyLa and SeAx cells. It further reduced p-p38, p-ERK, p-AKT, and p-STAT in MyLa cells, while it decreased p-ERK and p-STAT in SeAx cells in CAM tumors. Our data demonstrated that the CAM assay could be employed as a preclinical in vivo model in CTCL for pharmacological testing. In agreement with previous in vitro data, the combination of Resminostat and Ruxolitinib was shown to exert antitumor effects in CTCL in vivo.

Ethnobotanical study of medicinal plants used by local communities of Damot Woyde District, Wolaita Zone, Southern Ethiopia

Abstract. Megersa M, Woldetsadik S. 2022. Ethnobotanical study of medicinal plants used by local communities of Damot Woyde District, Wolaita Zone, Southern Ethiopia. Nusantara Bioscience 14: 10-24. Humans have used traditional medicines mainly of plant origins to treat diseases. Early humans faced a tremendous challenge when searching for natural products used as medicines. This study reports on an ethnobotanical study that focused on the traditional medicinal plants used by local communities to treat human and livestock diseases. An ethnobotanical study on medicinal plants was conducted from February 2020 to October 2020. This involved semi-structured interviews, field observations, market surveys and group discussions with informants to document information on the use and management of medicinal plants by the people of Damot Woyde District. Fifty-seven medicinal plant species belonging to 31 families were collected, which are used by the study area inhabitants to treat various diseases in humans and livestock. The leading family was Asteraceae which was represented by 7 species (12.3%), followed by Rutaceae (6 species, 10.5%) and Solanaceae (5 species, 9%). Of the 57 medicinal plants collected, 36 (63.2%) were used to treat human ailments only, while 6 (10.5%) plant species were used to treat livestock ailments only and 15 (26.31%) were used to treat both human and livestock ailments. Herbs constituted the largest number of 22 species (38.6%), followed by shrubs 18 species (31.6%), trees 15 species (26.3%) and climbers 2 species (3.51%). Leaves (31.3%) were the most commonly used plant parts of preparing traditional remedies in the study area. Oral administration was the predominant mode of administration accounting for 71%. Preference ranking analysis revealed that Allium sativum L. was the most preferred plant species for treating the common cold. When the direct matrix ranking was analyzed, Croton macrostachyus Hochst. ex. Del. was the most commonly used medicinal plant for various purposes. Our finding revealed that plant species' use plays a vital role in treating human and animal diseases in Damot Woyde District. Phytochemical and pharmacological tests are recommended mainly on frequently used medicinal plants.

Colorectal cancer on a dish: exploring the 3D-sphere culture of primary colorectal cancer cells from an Indonesian perspective

Background : Colorectal Cancer (CRC) is one of the deadliest types of cancer and has emerged as one of Indonesia's most devastating diseases. The growing number of colorectal cancer cases is frequently undiagnosed until the disease has progressed to a metastatic stage. This issue has lasted for years, limiting therapy options and resulting in a bad prognosis for the majority of patients. Thus, the purpose of this work is to develop a CRC detection method for Indonesia and other low-middle income nations that integrates in vitro 3D culture, molecular analysis, and in silico analysis. Methods : Colorectal cancer biopsies were transported to the lab and underwent mechanical disaggregation and centrifuged at 300 x g for five minutes. Approximately 10,000 cells were seeded in each Nunc-Sphera 96-well plate (u-bottom) for the following 7 days in standard culture medium. The 3D-sphere was harvested and RNA was extracted afterwards. Molecular analysis was performed using qPCR and the Human Cancer Pathway Profiler. Protein interaction and pathway analysis were conducted using STRING and Reactome online tools. Results : Following initial seeding, primary CRC 3D-spheres were grown for 14–16 days. Gene profiling and in silico analyses suggest that CDC20, AURKA, and ACLY are expressed at lower levels than the positive control in the 3D-sphere. These markers have been implicated in metastasis, CRC proliferation, and as a drug target ligand. Conclusion : A combination of 3D culture, gene profiling, and in silico analysis is feasible to detect CRC for Indonesia and other low- and middle-income countries. A future possibility is to use minicolorectal cancer in a dish for ex vivo cancer modeling and pharmacological testing.

Discovery of the specific inhibitory effect of thiamphenicol on LPS-induced acute lung injury (ALI) in mice through virtual screening and biological evaluation

Acute lung injury (ALI) is an acute and persistent pulmonary inflammatory syndrome. Neutrophil elastase (NE) activity is increased during ALI to regulate lung inflammation and hydrolyze a variety of matrix proteins that subsequently cause lung injury. Small molecule compounds with pyrrolidine trans-lactam structure can bind NE and inhibit its activity, improving the symptoms of ALI in model animals. Based on this information, we conducted virtual screening of small molecule compounds with pyrrolidine trans-lactam structures that may have NE inhibitory activity in the compound libraries and combined with in-depth screening and molecular docking validation, the compound thiamphenicol (TAP) with potential NE inhibitory activity was finally obtained. In vivo experiments in mice with ALI showed that TAP could significantly reduce LPS-induced pulmonary edema, decrease microvascular permeability, improve pathological and ultrapathological damage in lung tissue, and maintain the normal structure and function of the alveolar-capillary endothelial barrier. It could also improve the degradation of vascular endothelial-cadherin (VE-cad). Pharmacological mechanism tests also demonstrated that TAP significantly inhibited NE activity in lung tissue of LPS-induced ALI mice, but did not affect the expression of NE protein. In addition, the application of TAP did not cause side effects of glucocorticoids in the conventional clinical treatment of ALI. We believe that this paper could provide a new structure for drugs used in the treatment of ALI, which will certainly contribute to the development of new chemical structures as drugs for the treatment of ALI.

Anti-Tumor Active Isopropylated Fused Azaisocytosine-Containing Congeners Are Safe for Developing Danio rerio as Well as Red Blood Cells and Activate Apoptotic Caspases in Human Breast Carcinoma Cells.

New isopropylated fused azaisocytosine-containing congeners (I–VI) have previously been reported as promising anticancer drug candidates, so further research on these molecules in the preclinical development phase is fully justified and necessary. For this reason, in the present paper, we assess the toxicity/safety profiles of all the compounds using Danio rerio and red blood cell models, and examine the effect of the most selective congeners on the activation of apoptotic caspases in cancer and normal cells. In order to evaluate the effect of each molecule on the development of zebrafish embryos/larvae and to select the safest compounds for further study, various phenotypic parameters (i.e., mortality, hatchability, heart rate, heart oedema, yolk sac utilization, swim bladder development and body shape) were observed, and the half maximal lethal concentration, the maximal non-lethal concentration and no observed adverse effect concentration for each compound were established. The effect of all the isopropylated molecules was compared to that of an anticancer agent pemetrexed. The lipophilicity-dependent structure–toxicity correlations were also determined. To establish the possible interaction of the compounds with red blood cells, an ex vivo hemolysis test was performed. It was shown that almost all of the investigated isopropylated congeners have no adverse phenotypic effect on zebrafish development during five-day exposure at concentrations up to 50 μM (I–III) or up to 20 μM (IV–V), and that they are less toxic for embryos/larvae than pemetrexed, demonstrating their safety. At the same time, all the molecules did not adversely affect the red blood cells, which confirms their very good hemocompatibility. Moreover, they proved to be activators of apoptotic caspases, as they increased caspase-3, -7 and -9 levels in human breast carcinoma cells. The conducted research allows us to select—from among the anticancer active drug candidates—compounds that are safe for developing zebrafish and red blood cells, suitable for further in vivo pharmacological tests.

Emerging Bioelectronic Strategies for Cardiovascular Tissue Engineering and Implantation.

Heart diseases are currently the leading cause of death worldwide. The ability to create cardiovascular tissue has numerous applications in understanding tissue development, disease progression, pharmacological testing, bio-actuators, and transplantation; yet current cardiovascular tissue engineering (CTE) methods are limited. However, there have been emerging developments in the bioelectronics field, with the creation of biomimetic devices that can intimately interact with cardiac cells, provide monitoring capabilities, and regulate tissue formation. Combining bioelectronics with cardiac tissue engineering can overcome current limitations and produce physiologically relevant tissue that can be used in various areas of cardiovascular research and medicine. This review highlights the recent advances in cardiovascular-based bioelectronics. First, cardiac tissue engineering and the potential of bioelectronic therapies for cardiovascular diseases are discussed. Second, advantageous bioelectronic materials for CTE and implantation and their properties are reviewed. Third, several representative cardiovascular tissue-bioelectronic interface models and the beneficial functions that bioelectronics can demonstrate in in vitro and in vivo applications are explored. Finally, the prospects and remaining challenges for clinical application are discussed.

Ambulatory electrocardiography, heart rate variability, and pharmacologic stress testing in cats with subclinical hypertrophic cardiomyopathy

The utility of ambulatory electrocardiography (AECG) to evaluate cats with subclinical hypertrophic cardiomyopathy (HCM) for arrhythmias and heart rate variability (HRV) is not well defined but may provide information regarding risk stratification. This prospective study used AECG to evaluate ectopy and HRV in subclinical HCM cats compared to healthy controls and is the first to implement a pharmacologic cardiac stress test. Twenty-three purpose-bred, Maine coon cross cats (16 HCM, 7 control) underwent 48-h of continuous AECG. Terbutaline (0.2–0.3 mg/kg) was administered orally at 24 and 36 h. Heart rate, ectopy frequency and complexity and HRV parameters, including standard deviation of normal R-R intervals (SDNN), were compared pre-terbutaline and post-terbutaline and across phenotype, genotype and sex. Genotype for an HCM-causative mutation was significantly associated with the frequency of supraventricular (P = 0.033) and ventricular (P = 0.026) ectopy across all cats. Seven HCM cats and zero healthy cats had a sinus arrhythmia. Mean heart rate was significantly higher post-terbutaline (p < 0.0001). HCM cats had significantly greater HRV compared to controls (SDNN: p = 0.0006). Male cats had significantly higher HRV (SDNN: p = 0.0001) and lower mean heart rates (p = 0.0001). HRV decreased post-terbutaline (SDNN: p = 0.0008) and changes in HRV observed between sexes were attenuated by terbutaline.

Regadenoson stress echocardiography: a road ahead

Abstract Funding Acknowledgements Type of funding sources: None. Introduction Drugs use in stress echocardiography (SE) have important limitations due to side effect and contraindications. Regadenoson (R) is emerging as a selective coronary vasodilator for pharmacological stress tests. Several studies have analyzed its diagnostic accuracy, but its prognostic value has not yet been studied. Purpose The objective is to assess safety and short-medium term prognosis in patients (p) who underwent SE with R. Methods Retrospective observational study. 126 patients were included from 2017 to 2020 and SE to study the detection of myocardial ischemia with R were performed. A positive SE test were consider if presented at least one of the follow: segmental changes in myocardial contractility, electrocardiography ischaemic changes or ischaemic symtomps. Adverse events (AE) were collected. To assess prognosis, we evaluated cardiovascular events (CV) with a median follow-up of 23 months, considering heart failure, angina, myocardial infarction (AMI) o cardiovascular death. We compare negative result SE test group (NG) with group of positive results (PG). Results Sixteen (13%) patients, presented AE, generally mild, with nausea in 4 patients. Two patients (1.6%) required the use of aminophylline for severe AE type bronchospasm. Nineteen studies (15%) were positive: 17 (89%) showed echocardiographically positive results, 5 of them also had clinical or electrocardiographic positivity, one (5%) was considered positive only by electrocardiographic criteria and another by clinical criteria. There were no CV deaths at follow-up. In the NG, 6 events were observed versus 2 in the PG (6 vs 11%). Time to event of the NG was a median of 90 [3-600] days versus 110 [40-180] days in the PG. The most frequent event was HF (4 NG vs 1 PG), coronary angiography was performed in only 2 patients with HF of the NG and did not show significant lesions. In the NG there were 2 IAM (2 vs 0), one had severe stenosis in the right coronary artery and the other several moderate diffuse stenosis. There was no events for angina in the NG (0 vs 1). The annual cumulative incidence was 11% in the PG compared to 4% in the NG. Conclusions Stress echocardiography with regadenoson is a safe test with a low serious adverse events rate. As concern as prognosis, In our sample, NG presented a lower proportion of events with a lower annual cumulative incidence than PG and also in a 23-month follow-up. Abstract Figure.

Expression of T-Type Voltage-Gated Calcium Channel in the Cilia of Human Nasal Epithelial Cells

Introduction: The mucociliary transport function of the airway epithelium is largely dependent on ciliary beating. The control signal of ciliary beating is thought to be intracellular Ca2+. We herein investigated the expression of T-type voltage-gated calcium channel (VGCC), a generator of intracellular Ca2+ oscillation, in the human nasal mucosa. Methods: The inferior turbinate was collected from patients with chronic hypertrophic rhinitis. The expression of T-type VGCC α1 subunits was examined by immunohistochemistry, transmission immunoelectron microscopy, Western blot, and real-time reverse transcription-polymerase chain reaction (RT-PCR). Participation of T-type VGCC in the ciliary beat regulation was examined by pharmacological inhibition tests using specific blockers of T-type VGCC in ex vivo measurements of the ciliary beat frequency (CBF) and ATP release and in intracellular Ca2+ imaging of isolated ciliated cells. Results: Immunohistochemical staining showed the expressions of T-type VGCC α1 subunits, Cav3.1 and Cav3.3, on the surface of the epithelial cells. At the ultrastructural level, immunoreactivity for Cav3.1 was localized on the surface of the cilia, and that for Cav3.3 was localized in the cilia and at the base of the cilia. The existence of Cav3.1 and Cav3.3 was confirmed at the protein level by Western blot and at the transcriptional level by real-time RT-PCR. Specific blockers of T-type VGCC, mibefradil and NNC 55-0396, significantly inhibited CBF. These blockers also inhibited a CBF increase induced by 8-bromo-cAMP/8-bromo-cGMP and significantly lowered the intracellular Ca2+ level of isolated ciliated cells in a time-dependent manner. On the other hand, the ATP release from the nasal mucosa was not changed by mibefradil or NNC 55-0396. Conclusion: These results indicate that T-type VGCC α1 subunits, Cav3.1 and Cav3.3, exist at the cilia of the nasal epithelial cells and participate in the regulation of ciliary beating and that these channels act downstream of cAMP/cGMP.

Identification of Silent Myocardial Ischemia in Patients with Long-Term Type 1 and Type 2 Diabetes

(1) Background: This study aimed to analyze epidemiological data to identify risk factors for silent myocardial ischemia in patients with long-term type 1 and type 2 diabetes. (2) Methods: An analysis was performed on 104 patients with long-term type 1 and type 2 diabetes who had not previously been diagnosed with cardiovascular disease. During hospitalization, patients were subjected to a standard ECG exercise test on a treadmill. If the test could not be performed or the result was uncertain, a pharmacological exercise test with dobutamine was performed. In the case of a positive exercise ECG test or a positive dobutamine test, the patient underwent coronary angiography. (3) Results: Atherosclerotic lesions were found in 24 patients. Patients with silent ischemia were significantly older and had a lower mean left ventricular ejection fraction and a higher incidence of carotid atherosclerosis. The presence of microvascular complications did not increase the risk of silent ischemia. (4) Conclusions: Silent heart ischemia is more common in type 2 than type 1 diabetes. Predisposing factors include older age, coexistence of carotid atherosclerosis, lower left ventricular ejection fraction, and smoking in patients with type 1 diabetes. Concomitant microvascular complications are not a risk factor.

Design, Synthesis, Molecular Docking, and Evaluation Antioxidant and Antimicrobial Activities for Novel 3-Phenylimidazolidin-4-One and 2-Aminothiazol-4-One Derivatives.

On our way to discovering and developing compounds that have an antioxidant impact compared to ascorbic acid and other biological activities, we designed, synthesized, and evaluated a new series of heterocyclic moieties drugs (1–11) as antioxidants and antimicrobial agents. As starting moieties, these new candidates were derived from two promising heterocyclic compounds, imidazoldin-4-one and thiazol-4-one. Firstly, diphenylimidazol 1 was obtained because of the cyclo condensation one-pot ternary reaction of urea, aniline, and chloroacetic acid under thermal conditions. Out of this starting compound, we could design and create new vital rings such as purine and triazine as in compounds 5 and 6, respectively. Secondly, the start thiazole derivative 7 was obtained from the intermolecular cyclization of thiourea, chloroacetic acid, p-nitobezaldehyde in the presence of sodium acetate. We synthesized various derivatives from this second starting compound 7 by being subjected to different reagents such as aniline, phenylenediamine, phenylhydrazine, and barbituric acid to yield 8, 9, 10, and 11, respectively. Using ascorbic acid as the standard compound, the pharmacological testing for antioxidant activity assessment of the produced derivatives was evaluated against ABTS (2,20-azinobis (3-ethylbenzothiazoline-6-sulfonic acid). Candidate 6 exhibited the best activity as an antioxidant agent compared to ascorbic acid as a reference compound. Moreover, all compounds were evaluated as antimicrobial agents against a series of bacteria and fungi. Among all synthesized compounds, compound 6 achieved high efficiency against two types of fungi and four kinds of bacteria, as Clotrimazole and Ampicillin were used as the reference agents, respectively. All chemical structures of the novel synthesized candidates were unequivocally elucidated and confirmed utilizing spectroscopical and elemental investigations.

Advancing research in autoimmune neuromuscular disorders

A new Series on autoimmune neuromuscular disorders in The Lancet Neurology comprises three Reviews 1 Huijbers MG Marx A Plomp JJ Le Panse R Phillips WD Advances in the understanding of disease mechanisms of autoimmune neuromuscular junction disorders. Lancet Neurol. 2022; 21: 163-175 Google Scholar , 2 Verschuuren JJGM Palace J Murai H Tannemaat M Kaminski HJ Bril V Advances and ongoing research in the treatment of autoimmune neuromuscular junction disorders. Lancet Neurol. 2022; 21: 189-202 Google Scholar , 3 Punga AR Maddison P Heckmann JM et al. Epidemiology, diagnostics, and biomarkers of autoimmune neuromuscular junction disorders. Lancet Neurol. 2022; 21: 176-188 Google Scholar that altogether provide a broad overview of the current status of, and ongoing research into, the causes and treatment of myasthenia gravis and Lambert-Eaton myasthenic syndrome. The major contribution of this Series is the comprehensive information that they provide about disease mechanisms, 1 Huijbers MG Marx A Plomp JJ Le Panse R Phillips WD Advances in the understanding of disease mechanisms of autoimmune neuromuscular junction disorders. Lancet Neurol. 2022; 21: 163-175 Google Scholar new drugs in development, 2 Verschuuren JJGM Palace J Murai H Tannemaat M Kaminski HJ Bril V Advances and ongoing research in the treatment of autoimmune neuromuscular junction disorders. Lancet Neurol. 2022; 21: 189-202 Google Scholar and known and potential biomarkers. 3 Punga AR Maddison P Heckmann JM et al. Epidemiology, diagnostics, and biomarkers of autoimmune neuromuscular junction disorders. Lancet Neurol. 2022; 21: 176-188 Google Scholar Advances and ongoing research in the treatment of autoimmune neuromuscular junction disordersMyasthenia gravis and Lambert-Eaton myasthenic syndrome are antibody-mediated autoimmune diseases of the neuromuscular junction that usually present with weakness in ocular muscles and in proximal muscles of the limb and trunk. Prognosis regarding muscle strength, functional abilities, quality of life, and survival is generally good. However, some patients do not respond to treatment. Symptomatic drugs, corticosteroids, and steroid-sparing immunosuppressive drugs remain the cornerstone of treatment. Full-Text PDF Advances in the understanding of disease mechanisms of autoimmune neuromuscular junction disordersMuscle weakness and fatigue are the hallmarks of autoimmune neuromuscular junction disorders. Although a plethora of immunosuppressive treatments exist, no cure is available to date and many patients are left with debilitating muscle weakness. Recent advances in the understanding of the structure and function of the neuromuscular junction, and the development of novel in vitro and in vivo models, have been instrumental in unravelling the pathophysiology of these autoimmune diseases. These advances are providing the rationale for the development of new therapeutic strategies. Full-Text PDF Epidemiology, diagnostics, and biomarkers of autoimmune neuromuscular junction disordersAutoimmune neuromuscular junction disorders are rare. However, myasthenia gravis is being increasingly recognised in people older than 50 years. In the past 5–10 years, epidemiological studies worldwide suggest an incidence of acetylcholine receptor antibody-positive myasthenia gravis of up to 29 cases per 1 million people per year. Muscle-specific tyrosine kinase antibody-positive myasthenia gravis and Lambert-Eaton myasthenic syndrome are about 20 times less common. Several diagnostic methods are available for autoimmune neuromuscular junction disorders, including serological antibody, electrophysiological, imaging, and pharmacological tests. Full-Text PDF

Evaluation of the ethanolic extract of Myconia albicas (Old Cinnamon) in the alternative model of anxiety in zebrafish

Myconia albicans is the most abundant genus of the melastomaceous botanical family. In Brazil, it is known as "Canela de Velho", and is used mainly in inflammatory processes. Due to the scarcity of scientific works that report its pharmacological effect, the objective of this study was to perform its chemical characterization, toxicity, and the possible effect on the nervous system, using the experimental model Zebrafish (Dario regio). The organic extract of ethyl acetate (EtOAc) was elaborated through the dry leaves of the cinnamon tree of old. The EtOAc sample was submitted to preliminary phytochemical screening and showed the presence of phenols, flavanoids of the flavones type, flavonols and xanthones, triterpenoids, and alkaloids. Then, flavonoid compounds (369.58±0.074mgEQ/g) and total phenols (81.48±0.005mgEAG/g) were determined. EtOAc was evaluated for its toxicity, with Artemia salina, which showed low toxicity in all the studied doses. In the pharmacological action tests, the doses of 3, 10, or 30 mg/mL were used; 20 µL; oral (v.o.) or intraperitoneal (i.p.). The locomotor activity was evaluated in the open field test, which showed a decrease in the fish. And the anxiolytic activity, in the light &amp; dark preference test, showed that EtOAc at 30 mg/mL, when administered intraperitoneally (IP), has an anxiolytic effect similar to Diazepam IP control. Thus the EtOAC of M. albicans showed the presence of total phenols and several flavonoids, with low toxicity, in which the highest dose presented anxiolytic effect in Zebrafish.

Indigenous Knowledge on Medicinal Flora Utilized by the Traditional Healers in Mappillaiyurani Village, Tuticorin District, Tamil Nadu, India

Objectives: The purpose of this study is to document the indigenous knowledge of medicinal flora utilized by the traditional healers of Mappillaiyurani, Tuticorin district to establish the relative importance, consensus and scope of all medicinal flora used. Methods: Indigenous knowledge of medicinal plants was executed through a questionnaire with traditional healers who are custodians of the knowledge about herbal medicine. Local floras of the state had been systematically followed to identify the plants. Results: A total of 42 medicinal plant species from 26 families were documented. The leaves were the most commonly used plant part, and herbal remedies were mostly prepared in the form of a decoction or paste and consumed internally. Respiratory diseases are the most common diseases reported by traditional healers in the study area. Conclusion: This documented report indicated that indigenous medicinal plants are a good source of plant-based safe drugs. Moreover, additional pharmacological tests are required to establish the efficacy and potency of the plants as medicine.

Screening and Antidiarrheal Activity Testing of Sembung Rambat (Mikania micrantha) Leaves

BACKGROUND: The prevalence of diarrhea in Indonesia is exceedingly high. In consequence, it causes an increment of morbidity and mortality rate in toddlers. Furthermore, every year more than 1.3 billion toddlers suffer from diarrhea, and 3.2 million experience death from this disease. Based on empirical evidence in Pelita Jaya, Pesisir Barat of Lampung, Sembung Rambat (Mikania micrantha) leaves are commonly used to treat diarrhea, rheumatism, and bleeding wounds. In addition, alkaloids are one of the compounds contained in sembung rambat leaves that can reduce or inhibit the disposal of waste substances from the body or have antidiarrheal properties. AIM: The research aims to prove the potential effect of sembung rambat (M. micrantha) leaf extract as an antidiarrheal agent by screening and conducting a pharmacological test on male Swiss mice. METHODS: The phytochemical qualitative analysis was employed to screen the M. micrantha leaf extract. Moreover, the antidiarrheal effect was tested using the intestinal transit method of diarrhea in mice induced by Oleum Ricini. RESULTS: The phytochemical screening results show that there are the secondary metabolites of alkaloids, tannins, flavonoids, and saponins contained in the extract. The measurement of the marker length on the intestine length (ratio) after t = 65 min exhibits that the ethanol extract of sembung rambat (M. micrantha) leaf contains an antidiarrheal activity with a dose variation of 100 mg/kg BW, 150 mg/kg BW, and 200 mg/kg BW. CONCLUSIONS: Ethanol extract of sembung rambat (M. micrantha) leaves at various doses of 100 mg/kg BW, 150 mg/kg BW, and 200 mg/kg BW possesses antidiarrheal activities. The most effective dose is 200 mg/kg BW as it performed the closest ratio to the positive control group.

Study on the Mechanism of Zhenwu Decoction in Decreasing the Fats in Non-Alcoholic Fatty Liver Disease Using System Pharmacology and Cytological Validation Test

Purpose of this study was to screen the active compounds of Zhenwu decoction and their gene targets in the treatment of non-alcoholic fatty liver disease by network pharmacology and cytological validation test. The candidate compounds and related targets of Zhenwu decoction were obtained from traditional Chinese medicine system pharmacology database and PharmMapper. The non-alcoholic fatty liver disease-related genes were obtained from online Mendelian inheritance in man, GeneCards and DisGeNET databases. Molecular docking was carried out to simulate the binding affinities between potential core compounds and key target genes, and the chemical constituents of Zhenwu decoction were analyzed and identified by Ultra-Performance Liquid Chromatography-Quadrupole Time-of-Flight Mass Spectrometry technology. Therapeutic effect and mechanism of Zhenwu decoction on non-alcoholic fatty liver disease were validated using in vitro analysis. The protein-protein interaction network analysis identified the albumin, protein kinase B1, epidermal growth factor receptor, caspase 3 and peroxisome proliferator activated receptor gamma as the key target genes. Gene ontology annotation analysis showed that the Zhenwu decoctionnon- alcoholic fatty liver disease target genes were mainly involved in the response to steroid hormone and lipid catabolic process. The results of the Kyoto encyclopedia of genes and genomes pathway enrichment analysis were mainly related to the Forkhead box O signaling pathway and phosphatidylinositol 3-kinaseprotein kinase B signaling pathway. Paeoniflorgenone had the highest binding affinities for albumin, protein kinase B1, epidermal growth factor receptor, caspase 3 and peroxisome proliferator activated receptor gamma. The in vitro experiment showed that 19.5 mg/ml (p≤0.05) and 39 mg/ml (p≤0.01) of Zhenwu decoction could reduce lipid accumulation. Real-time quantitative polymerase chain reaction analyses revealed that Zhenwu decoction induced down-regulation of epidermal growth factor receptor messenger ribonucleic acid levels and up-regulation of heat shock protein 90 alpha family class a member 1, mitogen-activated protein kinase 1 and phosphatidylinositol 3-kinase messenger ribonucleic acid levels.

A high-value-added application of the stems of Rheum palmatum L. as a healthy food: the nutritional value, chemical composition, and anti-inflammatory and antioxidant activities.

Rhubarb has edible stems or stalks. In this paper, we investigated the nutritional value, chemical composition, and bioactivities of Rheum palmatum stems (SRP) and analyzed the mode of action. SRP exhibited biosafety and had nutritional value, with abundant essential amino acids and minerals. Based on network pharmacology and western blot tests, we found that it showed anti-inflammatory activity via the PI3K-Akt-mediated NF-κB pathway. Out of 20 compounds identified using UPLC-ESI-Q-TOF/MS analysis, cirsiliol and hydrangenol were active compounds and they inhibited NO production in RAW264.7 cells induced by LPS. The alleviation of an inflammatory response is combined with a decrease in oxidative stress, and SRP showed antioxidant activity via attenuating antioxidant enzymes, scavenging free radicals, improving the mitochondrial membrane potential, and decreasing the reactive oxygen species level. These results indicated that SRP, with abundant flavonoids and a good nutritional composition, could be used as a dietary supplement for food applications.

Molecular mechanism of Ganoderma against gastric cancer based on network pharmacology and experimental test

This study aims to explore the molecular mechanism of Ganoderma against gastric cancer based on network pharmacology, molecular docking, and cell experiment. The active components and targets of Ganoderma were retrieved from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP), and gastric cancer-related targets from GeneCards and Online Mendelian Inheritance in Man(OMIM). The protein-protein interaction(PPI) network of the common targets was constructed with STRING, followed by Gene Ontology(GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis of the common genes based on Bioconductor and R language. The medicinal-disease-component-target network and medicinal-disease-component-target-pathway network were established by Cytoscape. Molecular docking was performed between β-sitosterol(the key component in Ganoderma) and the top 15 targets in the PPI network. Cell experiment was performed to verify the findings. A total of 14 active components and 28 targets of Ganoderma were retrieved, and the medicinal and the disease shared 25 targets, including caspase-3(CASP3), caspase-8(CASP8), caspase-9(CASP9), and B-cell lymphoma-2(BCL2). The common targets involved 72 signaling pathways and apoptosis and p53 signaling pathway may play a crucial role in the effect of Ganoderma against gastric cancer. β-sitosterol had strong binding activity to the top 15 targets in the PPI network. The in vitro cell experiment demonstrated that β-sitosterol inhibited gastric cancer AGS cell proliferation by inducing cell apoptosis and cell cycle arrest in the S phase, which might be related to the regulation of the p53 pathway. This study shows the multi-component, multi-target, and multi-pathway characteristics of Ganoderma against gastric cancer, which lays a scientific basis for further research on the molecular mechanism.

CATALASE CRS-262T GENE POLYMORPHISM AND CHANGES IN VENTILATION LUNG CAPACITY IN CHILDREN-RESIDENTS OF RADIOACTIVELY CONTAMINATED TERRITORIES.

to determine the association of catalase С-262Т gene polymorphism with the presence of bronchial hyper-reactivity in children living in radioactively contaminated territories. There were examined school-age children-residents of radioactively contaminated territories (RCT), who did not have clinical signs of respiratory pathology. Catalase (CAT) С-262Т gene deletion polymorphism was studied in the molecular genetic laboratory of the State Institution «Reference Center for Molecular Diagnostic of Public Health Ministry of Ukraine». Determination of the polymorphic variant by the catalase С-262Т gene was performed by Polymerase Chain Reaction (PCR) using specific oligonucleotide primers, followed by Restriction Fragment Length Polymorphism (RFLP) analysis. The CAT С-262Т gene polymorphism in children living in RCT was compared with that in the reference group of practically healthy individuals. Ventilation lung capacity was performed by computer spirometry according to the analysis of the loop «the flow-volume». A pharmacological inhalation test with a bronchodilator that acts on β2-adrenergic receptors of the lungs was used to detect early changes in the ventilatory capacity of the lungs - bronchial hyperreactivity. Comparative analysis showed that in the presence of bronchial hyperreactivity in children living in RCT, the CT genotype was more common than in children without bronchial hyperreactivity, and the frequency of the CC genotype was correspondingly reduced. There was a trend towards a decrease in the frequency of the TT genotype. An analysis of the frequency distribution of allelic variants of the CAT С-262Т gene polymorphism in children living in the RCT revealed a tendency to increase in the frequency of the T-allele and according to the decrease in the frequency of C-allele in the presence of bronchial hyperreactivity.Сonclusions. Thus, among children living in RCT, CT-homozygotes of CAT С-262Т gene polymorphism had bronchial hyperreactivity probably more often than CC-heterozygotes. In the presence of bronchial hyperreactivity, there was a trend towards an increase in the frequency of the T-allele and, accordingly, a decrease in the frequency of the C-allele.

Diosgenin: an important natural pharmaceutical active ingredient

Diosgenin is a steroidal saponin isolated from dioscorea opposita, which has been proven to have anti-cancer, anti-inflammatory, anti-thrombotic and immune system modulating effects in a large number of pharmacological tests. Diosgenin is also one of the important raw materials for the synthesis of steroidal drugs, and structural modification of diosgenin to obtain diosgenin derivatives can not only improve the solubility and dissolution, but also enhance their pharmacological effects. This paper reviews the synthetic reactions, pharmacological effects of diosgenin and its derivatives, aiming to provide detailed information for the further development and application of diosgenin, which is important to improve the utilization value of diosgenin.