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A special issue of Pharmaceuticals (ISSN 1424-8247). This special issue belongs to the section "Natural Products".

Deadline for manuscript submissions: 28 February 2025 | Viewed by 23340

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Prof. Dr. Alexander George Panossian

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Phytomed AB, 58344 Vastervick, Sweden
Interests: network pharmacology; neuroendocrine-immune system; stress; ageing; infalmmation; adaptogens
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Special Issue Information

Dear Colleagues,

This Special Issue is aimed to highlight recent findings on the relationship between natural products (purified compounds, plant extracts, herbal medicines, complex formulations) and pharmacological activity, including specific and pleiotropic action.

We encourage authors to submit a wide range of studies, including molecular mechanisms of multitarget action based on network pharmacology approach and clinical trials.

Prof. Dr. Alexander George Panossian
Guest Editor

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Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Pharmaceuticals is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2900 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

natural compounds
plant extracts
structure-activity relationship
network pharmacology
multitarget effects
synergy of complex preparations
pleiotropic activity

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16 pages, 5109 KiB

Open AccessArticle

Zanthoxylum piperitum Benn. Attenuates Monosodium Urate-Induced Gouty Arthritis: A Network Pharmacology Investigation of Its Anti-Inflammatory Mechanisms

by Sung Wook Kim, Soo Hyun Jeong, Jong Uk Kim, Mi Hye Kim, Wonwoong Lee, Cheol-Jung Lee, Tae Han Yook and Gabsik Yang

Pharmaceuticals 2025, 18(1), 29; https://doi.org/10.3390/ph18010029 - 29 Dec 2024

Viewed by 708

Abstract

Background: Monosodium urate crystal accumulation in the joints is the cause of gout, an inflammatory arthritis that is initiated by elevated serum uric acid levels. It is the most prevalent form of inflammatory arthritis, affecting millions worldwide, and requires effective treatments. The necessity for alternatives with fewer side effects is underscored by the frequent adverse effects of conventional therapies, such as urate-lowering drugs. IL-1β is a potential therapeutic target due to its significant role in the inflammatory response induced by MSU. Zanthoxylum piperitum Benn. (ZP), a shrub that possesses antibacterial, antioxidant, and anti-inflammatory properties, has demonstrated potential in the treatment of inflammatory conditions. Methods: For anti-inflammatory properties of ZP, Raw264.7 cell stimulated LPS were treated ZP and using RNA-seq with Bone marrow derived macrophage, we observed to change inflammatory gene. Pharmacological networks were conducted to select target gene associated with ZP. For in vivo, mice were injected MSU in footpad for induce gouty arthritis model. The components of ZP were analyzed using GC-MS, and distilled extracts of ZP (deZP) were prepared. Results: In vitro, deZP decreased inflammatory cytokines. However, in vivo, it also decreased paw thickness and IL-1β levels. The anti-inflammatory effects of deZP are believed to be mediated through the NLRP3 inflammasome pathway, as indicated by RNA sequencing and network pharmacology analyses. Conclusions: ZP has an anti-inflammatory effect and regulation of the NLRP3 inflammasome in vitro and in vivo. Further research, including clinical trials, is required to confirm the safety of deZP, determine the optimal dosing, and evaluate its long-term effects. Full article

(This article belongs to the Special Issue Network Pharmacology of Natural Products)

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24 pages, 3209 KiB

Open AccessArticle

Multi-Omics Analysis in Mouse Primary Cortical Neurons Reveals Complex Positive and Negative Biological Interactions Between Constituent Compounds of Centella asiatica

by Steven R. Chamberlin, Jonathan A. Zweig, Cody J. Neff, Luke Marney, Jaewoo Choi, Liping Yang, Claudia S. Maier, Amala Soumyanath, Shannon McWeeney and Nora E. Gray

Pharmaceuticals 2025, 18(1), 19; https://doi.org/10.3390/ph18010019 - 27 Dec 2024

Viewed by 580

Abstract

Background: A water extract of the Ayurvedic plant Centella asiatica (L.) Urban, family Apiaceae (CAW), improves cognitive function in mouse models of aging and Alzheimer’s disease and affects dendritic arborization, mitochondrial activity, and oxidative stress in mouse primary neurons. Triterpenes (TT) and caffeoylquinic acids (CQA) are constituents associated with these bioactivities of CAW, although little is known about how interactions between these compounds contribute to the plant’s therapeutic benefit. Methods: Mouse primary cortical neurons were treated with CAW or equivalent concentrations of four TT combined, eight CQA combined, or these twelve compounds combined (TTCQA). Treatment effects on the cell transcriptome (18,491 genes) and metabolome (192 metabolites) relative to vehicle control were evaluated using RNAseq and metabolomic analyses, respectively. Results: Extensive differentially expressed genes (DEGs) were seen with all treatments, as well as evidence of interactions between compounds. Notably, many DEGs seen with TT treatment were not observed in the TTCQA condition, possibly suggesting CQA reduced the effects of TT. Moreover, additional gene activity seen with CAW as compared to TTCQA indicates the presence of additional compounds in CAW that further modulate TTCQA interactions. Weighted Gene Correlation Network Analysis (WGCNA) identified 4 gene co-expression modules altered by treatments that were associated with extracellular matrix organization, fatty acid metabolism, cellular response to stress and stimuli, and immune function. Compound interaction patterns were seen at the eigengene level in these modules. Interestingly, in metabolomics analysis, the TTCQA treatment saw the highest number of changes in individual metabolites (20), followed by CQA (15), then TT (8), and finally CAW (3). WGCNA analysis found two metabolomics modules with significant eigenmetabolite differences for TT and CQA and possible compound interactions at this level. Conclusions: Four gene expression modules and two metabolite modules were altered by the four treatment types applied. This methodology demonstrated the existence of both negative and positive interactions between TT, CQA, and additional compounds found in CAW on the transcriptome and metabolome of mouse primary cortical neurons. Full article

(This article belongs to the Special Issue Network Pharmacology of Natural Products)

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28 pages, 9041 KiB

Open AccessArticle

Salvia miltiorrhiza and Its Compounds as Complementary Therapy for Dyslipidemia: A Meta-Analysis of Clinical Efficacy and In Silico Mechanistic Insights

by Min-Seong Lee, Han-Young Lee, Seung-Hyun Oh, Chang-Bum Kim, Ji-Han Kim, Seung-Hoon Yoo, Yeon-Joo Yoo, Su-Yeon Lee and Byung-Cheol Lee

Pharmaceuticals 2024, 17(11), 1426; https://doi.org/10.3390/ph17111426 - 24 Oct 2024

Viewed by 1173

Abstract

Background/Objectives: Dyslipidemia is a significant risk factor for atherosclerotic cardiovascular disease (ASCVD), a leading cause of death worldwide. Salvia miltiorrhiza Burge is widely used in East Asia for cardiovascular health, showing potential benefits in lowering cholesterol and reducing inflammation. Methods: This study systematically reviewed and conducted a meta-analysis of randomized controlled trials (RCTs) to assess the clinical effectiveness of Salvia miltiorrhiza in treating dyslipidemia. Moreover, network pharmacology and molecular docking analyses were performed to explore the mechanisms underlying the effects of Salvia miltiorrhiza. Results: The meta-analysis revealed that when Salvia miltiorrhiza is combined with statin therapy, it significantly enhances lipid profiles, including reductions in total cholesterol, low-density lipoprotein cholesterol (LDL-C), and triglycerides and improvements in high-density lipoprotein cholesterol (HDL-C), compared to statin therapy alone. The in silico analyses indicated that Salvia miltiorrhiza may influence key biological pathways, such as the PI3K/Akt, JAK/STAT, and HMGCR pathways, which are involved in inflammation, lipid metabolism, and the development of atherosclerosis. Conclusions: Salvia miltiorrhiza shows potential as a complementary therapy for dyslipidemia, offering additional lipid-lowering and anti-inflammatory benefits. Full article

(This article belongs to the Special Issue Network Pharmacology of Natural Products)

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21 pages, 9362 KiB

Open AccessArticle

Potential Anti-Obesity Effect of Hazel Leaf Extract in Mice and Network Pharmacology of Selected Polyphenols

by Jiarui Zhao, Aikebaier Alimu, Yvmo Li, Zhi Lin, Jun Li, Xinhe Wang, Yuchen Wang, Guangfu Lv, He Lin and Zhe Lin

Pharmaceuticals 2024, 17(10), 1349; https://doi.org/10.3390/ph17101349 - 9 Oct 2024

Viewed by 1250

Abstract

Background: Obesity is gradually becoming a widespread health problem, and treatment using natural compounds has seen an increasing trend. As a by-product of hazelnut, hazel leaf is usually disposed of as waste, but it is widely used in traditional and folk medicines around the world. Aim of this study: Based on previous studies, the effects of the regulation of lipid metabolism and the mechanism of hazel leaf polyphenol extraction obesity were investigated. Methods: In this study, a high-fat diet-fed mouse model of obesity and 3T3-L1 preadipocytes were established. The ameliorative effects of the hazel leaf polyphenol extract on obesity and the regulating lipid metabolisms were explored based on network pharmacology, gut microbiota, and molecular docking. Results: Network pharmacology showed that hazel leaf polyphenols may play a role by targeting key targets, including PPARγ, and regulating the PPAR signaling pathway. They significantly improved body weight gain, the liver index, and adiposity and lipid levels; regulated the gut microbiota and short-chain fatty acid contents; down-regulated the expression of lipid synthesis proteins SREBP1c, PPARγ, and C/EBP-α; and up-regulated the expression of p-AMPK in obese mice. They inhibited the differentiation of 3T3-L1 cells, and the expression of related proteins is consistent with the results in vivo. The molecular docking results indicated that gallic acid, quercetin-3-O-beta-D-glucopyranoside, quercetin, myricetin, and luteolin-7-O-glucoside in the hazel leaf polyphenol extract had strong binding activities with PPARγ, C/EBP-α, and AMPK. Conclusions: The results demonstrate that the hazel leaf polyphenol extract can improve obesity by regulating lipid metabolism, which provides a valuable basis for developing health products made from hazel leaf polyphenols in the future. Full article

(This article belongs to the Special Issue Network Pharmacology of Natural Products)

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29 pages, 7238 KiB

Open AccessArticle

Unveiling the Bioactive Efficacy of Cupressus sempervirens ‘Stricta’ Essential Oil: Composition, In Vitro Activities, and In Silico Analyses

by Eman Fikry, Raha Orfali, Nora Tawfeek, Shagufta Perveen, Safina Ghafar, Maher M. El-Domiaty and Azza M. El-Shafae

Pharmaceuticals 2024, 17(8), 1019; https://doi.org/10.3390/ph17081019 - 2 Aug 2024

Viewed by 1716

Abstract

Prior studies have extensively investigated the essential oil derived from the Mediterranean cypress, Cupressus sempervirens. However, the ‘Stricta’ variety, known for its ornamental value, has received less attention in terms of its oil composition and potential health benefits. The objective of this research was to comprehensively analyze the chemical components and medicinal properties of the essential oil extracted from C. sempervirens ‘Stricta’ (CSSLEO) grown in Egypt. Utilizing gas chromatography–mass spectrometry (GC–MS), the investigation identified 22 compounds within CSSLEO, with α-pinene and δ-3-carene being predominant, accounting for 96.01% of the oil. In vitro assays evaluated CSSLEO’s cytotoxic effects on cancer cell lines, revealing notable anticancer potential. Additionally, the oil displayed antidiabetic properties by impeding crucial enzymes involved in glucose metabolism. Complementary in silico network pharmacology and molecular docking studies provided insights into the possible interactions between CSSLEO’s key compounds and essential proteins and pathways in cancer treatment. The results underscored CSSLEO’s intricate composition and its promising applications in cancer prevention and diabetes management. The conclusions drawn from this research underscore the need for further investigation to validate CSSLEO’s clinical effectiveness and to gain a deeper understanding of its therapeutic mechanisms, with a view to harnessing its potential in oncology and endocrinology. Full article

(This article belongs to the Special Issue Network Pharmacology of Natural Products)

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17 pages, 3702 KiB

Open AccessEditor’s ChoiceArticle

Aronia Berry Extract Modulates MYD88/NF-kB/P-Glycoprotein Axis to Overcome Gemcitabine Resistance in Pancreatic Cancer

by Yuan Li, Caiming Xu, Haiyong Han, Silvia Pascual-Sabater, Cristina Fillat and Ajay Goel

Pharmaceuticals 2024, 17(7), 911; https://doi.org/10.3390/ph17070911 - 9 Jul 2024

Cited by 1 | Viewed by 2954

Abstract

Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal disease with poor survival rates, primarily due to the limited effectiveness of gemcitabine (Gem)-based chemotherapy, as well as the acquisition of chemotherapeutic resistance. Aronia berry extracts (ABEs), abundant in phenolic constituents, have been recently recognized for their anticancer properties as well as their encouraging potential to help overcome chemoresistance in various cancers. In the present study, we explored ABE’s potential to overcome Gem resistance in PDAC and identify specific growth regulatory pathways responsible for its anticancer activity. Through a series of in vitro experiments in gemcitabine-resistant (Gem-R) cells, we elucidated the synergistic interactions between Gem and ABE treatments. Using advanced transcriptomic analysis and network pharmacology, we revealed key molecular pathways linked to chemoresistance and potential therapeutic targets of ABE in Gem-R PDAC cells. Subsequently, the findings from cell culture studies were validated in patient-derived 3D tumor organoids (PDOs). The combination treatment of ABE and Gem demonstrated significant synergism and anticancer effects on cell viability, proliferation, migration, and invasion in Gem-R cells. Transcriptomic analysis revealed a correlation between the NF-Κb signaling pathway and Gem-R (p < 0.05), exhibiting a marked upregulation of MYD88. Additionally, MYD88 exhibited a significant correlation with the overall survival rates in patients with PDAC patients in the TCGA cohort (HR = 1.58, p < 0.05). The MYD88/NF-Κb pathway contributes to chemoresistance by potentially upregulating efflux transporters like P-glycoprotein (P-gp). Our findings revealed that the combined treatment with ABE suppressed the NF-Κb pathway by targeting MYD88 and reducing P-gp expression to overcome Gem resistance. Lastly, the combination therapy proved highly effective in PDOs in reducing both their number and size (p < 0.05). Our study offers previously unrecognized insights into the ability of ABE to overcome Gem resistance in PDAC cells through its targeting of the MYD88/NF-κb/P-gp axis, hence providing a safe and cost-effective adjunctive therapeutic strategy to improve treatment outcomes in PDAC. Full article

(This article belongs to the Special Issue Network Pharmacology of Natural Products)

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20 pages, 5214 KiB

Open AccessArticle

Efficacy of Kan Jang^® in Patients with Mild COVID-19: A Randomized, Quadruple-Blind, Placebo-Controlled Trial

by Levan Ratiani, Elene Pachkoria, Nato Mamageishvili, Ramaz Shengelia, Areg Hovhannisyan and Alexander Panossian

Pharmaceuticals 2023, 16(9), 1196; https://doi.org/10.3390/ph16091196 - 22 Aug 2023

Cited by 1 | Viewed by 2610

Abstract

Background and aim. This study aimed to assess the efficacy of the treatment of Kan Jang^®, a fixed combination of Andrographis paniculata (Burm. F.) Wall. ex. Nees and Eleutherococcus senticosus (Rupr. & Maxim.) Maxim extracts in patients with mild symptoms of COVID-19. Methods. One hundred and forty patients received six capsules of Kan Jang^® (n = 68, daily dose of andrographolides—90 mg) or placebo (n = 72) and supportive treatment (paracetamol) for 14 consecutive days in a randomized, quadruple-blinded, placebo-controlled, two-parallel-group design. The efficacy outcomes were the rate of cases turning to severe, the detection rate of coronavirus SARS-CoV-2 over the time of treatment, the duration, and the severity of symptoms (sore throat, runny nose, cough, headache, fatigue, loss of smell, taste, pain in muscles) in the acute phase of the disease. Other efficacy measures included improving cognitive and physical performance, quality of life, and the levels of inflammatory blood markers—interleukin 6 (IL-6), C-reactive protein, and D-dimer. Results. Kan Jang^® significantly (p < 0.05) reduced the rate of cases turning to severe (5.36%) compared to the placebo (17.86%) and decreased the detection rate of SARS-CoV-2 virus over the time of the treatment. The statistical difference in the rates of patients with clinical deterioration in the Kan Jang treatment and placebo control groups was significant (p = 0.0176) both in the 112 patients in the included-per-protocol (IPP) analysis and in the 140 patients in the intended-to-treat (ITT) analysis (p = 0.0236); the absolute risk reduction in cases thanks to the Kan Jang treatment was 12.5%, and the number we needed to treat with Kan Jang was 8. The patient’s recovery time (number of sick days at the home/clinic) was shorter in the Kan Jang group compared with the placebo group. The rate of attenuation of inflammatory symptoms in the Kan Jang^® group was significantly higher, decreasing the severity of cough, sore throat/pain, runny nose, and muscle soreness compared with the placebo group. Kan Jang^® significantly decreased the Wisconsin Upper Respiratory Symptoms scores compared to the placebo in the sample size of 140 patients. However, the relief of fatigue and headache and the decrease in IL-6 in the blood were observed only in a subset of 86 patients infected during the second three waves of the pandemic. Kan Jang^® significantly increased physical activity and workout; however, it did not affect cognitive functions (attention and memory), quality of life score, inflammatory marker D-dimer, and C-reactive protein compared with the placebo group. Conclusions. Overall, the results of this study suggest that Kan Jang^® is effective in treating mild and moderate COVID-19 irrespective of the SARS-CoV-2 variant of infection. Full article

(This article belongs to the Special Issue Network Pharmacology of Natural Products)

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24 pages, 2676 KiB

Open AccessReview

Unlocking the Green Gold: Exploring the Cancer Treatment and the Other Therapeutic Potential of Fucoxanthin Derivatives from Microalgae

by Fatouma Mohamed Abdoul-Latif, Ayoub Ainane, Ibrahim Houmed Aboubaker, Ali Merito Ali, Houda Mohamed, Pannaga Pavan Jutur and Tarik Ainane

Pharmaceuticals 2024, 17(7), 960; https://doi.org/10.3390/ph17070960 - 18 Jul 2024

Cited by 2 | Viewed by 2138

Abstract

Fucoxanthin, a carotenoid widely studied in marine microalgae, is at the heart of scientific research because of its promising bioactive properties for human health. Its unique chemical structure and specific biosynthesis, characterized by complex enzymatic conversion in marine organisms, have been examined in depth in this review. The antioxidant, anti-inflammatory, and anti-cancer activities of fucoxanthin have been rigorously supported by data from in vitro and in vivo experiments and early clinical trials. Additionally, this review explores emerging strategies to optimize the stability and efficacy of fucoxanthin, aiming to increase its solubility and bioavailability to enhance its therapeutic applications. However, despite these potential benefits, challenges persist, such as limited bioavailability and technological obstacles hindering its large-scale production. The medical exploitation of fucoxanthin thus requires an innovative approach and continuous optimization to overcome these barriers. Although further research is needed to refine its clinical use, fucoxanthin offers promising potential in the development of natural therapies aimed at improving human health. By integrating knowledge about its biosynthesis, mechanisms of action, and potential beneficial effects, future studies could open new perspectives in the treatment of cancer and other chronic diseases. Full article

(This article belongs to the Special Issue Network Pharmacology of Natural Products)

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27 pages, 5792 KiB

Open AccessReview

State-of-the-Art Review on Botanical Hybrid Preparations in Phytomedicine and Phytotherapy Research: Background and Perspectives

by Alexander Panossian, Terry Lemerond and Thomas Efferth

Pharmaceuticals 2024, 17(4), 483; https://doi.org/10.3390/ph17040483 - 10 Apr 2024

Cited by 10 | Viewed by 3629

Abstract

Background: Despite some evidence supporting the synergy concept, the commonly known assumption that combinations of several herbs in one formulation can have better efficacy due to additive or synergistic effects has yet to be unambiguously and explicitly studied. Study aim: The study aimed to reveal the molecular interactions in situ of host cells in response to botanical hybrid preparations (BHP) intervention and justify the benefits of implementing BHP in clinical practice. Results: This prospective literature review provides the results of recent clinical and network pharmacology studies of BHP of Rhodiola rosea L. (Arctic root) with other plants, including Withania somnifera (L.) Dunal (ashwagandha), (Camellia sinensis (L.) Kuntze (green tea), Eleutherococcus senticosus (Rupr. and Maxim.) Maxim. (eleuthero), Schisandra chinensis (Turcz.) Baill. (schisandra), Leuzea carthamoides (Willd.) DC., caffeine, Cordyceps militaris L., Ginkgo biloba L.(ginkgo), Actaea racemosa L. (black cohosh), Crocus sativus L. (saffron), and L-carnosine. Conclusions: The most important finding from network pharmacology studies of BHP was the evidence supporting the synergistic interaction of BHP ingredients, revealing unexpected new pharmacological activities unique and specific to the new BHP. Some studies show the superior efficacy of BHP compared to mono-drugs. At the same time, some a priori-designed combinations can fail, presumably due to antagonistic interactions and crosstalk between molecular targets within the molecular networks involved in the cellular and overall response of organisms to the intervention. Network pharmacology studies help predict the results of studies aimed at discovering new indications and unpredicted adverse events. Full article

(This article belongs to the Special Issue Network Pharmacology of Natural Products)

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81 pages, 15918 KiB

Open AccessSystematic Review

Efficacy and Key Materials of East Asian Herbal Medicine Combined with Conventional Medicine on Inflammatory Skin Lesion in Patients with Psoriasis Vulgaris: A Meta-Analysis, Integrated Data Mining, and Network Pharmacology

by Hee-Geun Jo, Hyehwa Kim, Eunhye Baek, Donghun Lee and Ji Hye Hwang

Pharmaceuticals 2023, 16(8), 1160; https://doi.org/10.3390/ph16081160 - 15 Aug 2023

Cited by 5 | Viewed by 3868

Abstract

Psoriasis is a chronic inflammatory disease that places a great burden on both individuals and society. The use of East Asian herbal medicine (EAHM) in combination with conventional medications is emerging as an effective strategy to control the complex immune-mediated inflammation of this disease from an integrative medicine (IM) perspective. The safety and efficacy of IM compared to conventional medicine (CM) were evaluated by collecting randomized controlled trial literature from ten multinational research databases. We then searched for important key materials based on integrated drug data mining. Network pharmacology analysis was performed to predict the mechanism of the anti-inflammatory effect. Data from 126 randomized clinical trials involving 11,139 patients were used. Compared with CM, IM using EAHM showed significant improvement in the Psoriasis Area Severity Index (PASI) 60 (RR: 1.4280; 95% CI: 1.3783–1.4794; p < 0.0001), PASI score (MD: −3.3544; 95% CI: −3.7608 to −2.9481; p < 0.0001), inflammatory skin lesion outcome, quality of life, serum inflammatory indicators, and safety index of psoriasis. Through integrated data mining of intervention data, we identified four herbs that were considered to be representative of the overall clinical effects of IM: Rehmannia glutinosa (Gaertn.) DC., Isatis tinctoria subsp. athoa (Boiss.) Papan., Paeonia × suffruticosa Andrews, and Scrophularia ningpoensis Hemsl. They were found to have mechanisms to inhibit pathological keratinocyte proliferation and immune-mediated inflammation, which are major pathologies of psoriasis, through multiple pharmacological actions on 19 gene targets and 8 pathways in network pharmacology analysis. However, the quality of the clinical trial design and pharmaceutical quality control data included in this study is still not optimal; therefore, more high-quality clinical and non-clinical studies are needed to firmly validate the information explored in this study. This study is informative in that it presents a focused hypothesis and methodology for the value and direction of such follow-up studies. Full article

(This article belongs to the Special Issue Network Pharmacology of Natural Products)

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Planned Papers

The below list represents only planned manuscripts. Some of these manuscripts have not been received by the Editorial Office yet. Papers submitted to MDPI journals are subject to peer-review.

Title: An Assessment of Antidepressant Like Potentiality of Phyllanthus emblica on Experimental Rodent Model of Depressive Disorder Under the Light of Numerous Behavioural Test Batteries
Author: Aktar
Highlights: Assessed psychological state of (test batteries) Stress given. Employing test batteries, severity of depression measured, Extract induced Using test batteries, severity of depression measured again. Extract effectively reverse the disturbed pathological state, severe to less severe or fully reversed. Such three-layered analysis has never been done in the same research previously.

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