Medicine Prescription Research Articles

Medicine Prescribing Practices and Prescription Errors Evaluations at Outpatient Department in Debre Berhan Comprehensive Specialized Hospital, Amhara Regional State, Ethiopia.

Background: Currently, irrational uses of medicines becoming global problem largely in developing countries like Ethiopia. Inappropriate prescribing is a major cause for poor treatment outcome and higher costs. Hence, this study was aimed to investigate medicine prescribing practice and prescription errors using WHO medicine-utilization core indicators. Methods: A hospital based retrospective cross sectional study design was used to evaluate prescribing practices and prescription errors from September to October, 2024 at the OPD pharmacy using systematic random sampling technique while a prospective approach was employed for facility indicators. Presence of potential drug-drug interactions (DDIs) were evaluated using Medscape Online Drug Interaction Checker. Data were analyzed using SPSS version 25 and interpreted as tables and figures. Results: A total of 1019 medicines were prescribed in 524 prescriptions and 81.6% (n = 832) were actually dispensed. The percentage of antibiotic, injections and medicine prescribed from Essential Drug List was 33.9% (n = 345),3.5% (n = 36) and 92.3% (n = 941) respectively. The most frequently prescribed class of medicine were antibiotics 33.9% (n = 345). 65.1% (n = 341) were ≥2 medicines and 8.3% (n = 85) had at least one potential DDIs. Among overall DDIs, the monitor closely and serious level was 60% (n = 51) and 11.8% (n = 10) respectively. The average prescription error was 4.3. Prescription errors due to failure to mention diagnosis was 40.6% (n = 213). Conclusion: Based on findings, the prescribing practices had defects to the optimum value recommended by WHO and showed high prescription errors. Antibiotics prescribing was the major problem in practice. Remarkable DDIs were observed in prescribed medicines. Therefore, designing and implementing policy to improve medicine use practice is highly indispensable.

The clinical efficacy of Lou's Wentong method in the treatment of lactational mastitis: A retrospective self-control study with 158 cases

BackgroundAs a common breast disease in lactating women, lactational mastitis is clinically manifested as painful breast lumps and systemic symptoms. To a certain extent, this disease may affect the physical and mental health of lactating women and even the breastfeeding of infants. ObjectiveThis study was conducted to evaluate the clinical efficacy of Lou's Wentong Method in the comprehensive treatment of lactational mastitis and its effects on relevant symptoms. MethodsA total of 158 patients who were diagnosed with lactational mastitis in The First Affiliated Hospital of Zhejiang Chinese Medical University from January 2022 to January 2024 were included in this study to conduct a retrospective analysis. The treatment method comprised oral administration of traditional Chinese medicine prescriptions guided by the concept of “Wentong zhi yong” (warming and dispersing therapy for treating abscesses), puncture and pus extraction for patients with abscess formation, and milk drainage for patients with milk stagnation. The clinical symptoms of these patients were collected according to the seven indexes in Lou's quantitative scorecard for evaluating the condition of canker sores before and after treatment. Additionally, the improvement of each symptom after treatment, the total effective rate of the clinical treatment, and the total symptom evaluation score before and after treatment were also analyzed. ResultsIt was demonstrated that there were significant differences in the overall distribution of different severity symptoms (skin erythema, breast pain, number of lumps, size of lumps, breast abscesses, and body temperature) at three time points (P < 0.01). The evaluation of these symptoms at different time points revealed that all symptoms were effectively mitigated over time. Meanwhile, the efficacy was assessed according to the efficacy index to evaluate the efficacy of Lou's Wentong Method in the comprehensive treatment of lactational mastitis. The results corroborated the pronounced efficacy of Lou's Wentong Method in treating lactational mastitis, exhibiting statistical significance (P < 0.01). ConclusionLou's Wentong Method has significant therapeutic effects in the comprehensive treatment of lactational mastitis. Furthermore, this method is conducive to alleviating the clinical symptoms of these patients as soon as possible and can reduce the pain of patients, thus promoting their recovery.

Mechanism of Chaihuang-Yishen formula to attenuate renal fibrosis in the treatment of chronic kidney disease: Insights from network pharmacology and experimental validation

Renal fibrosis represents a pivotal characteristic of chronic kidney disease (CKD), for which effective interventions are currently lacking. The Src kinase activates the phosphatidylinositol-3 kinases (PI3K)/Akt1 pathway to promote renal fibrosis, casting a promising target for anti-fibrosis treatment. Chaihuang-Yishen formula (CHYS), a traditional Chinese medicinal prescription, has a validated efficacy in the treatment of CKD, however, with the underlying mechanism unresolved. This study aimed to uncover the pharmacological mechanisms mediating the effect of CHYS in treating renal fibrosis using network pharmacology followed by experimental validation. The chemical compounds of CHYS were retrieved from the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database or published literature, followed by the prediction of their targets using SwissTargetPrediction software. Disease (CKD/renal fibrosis)-related targets were retrieved from the Genecards database. Protein-protein interaction (PPI) network was generated using the drug-disease common targets and visualized in Cytoscape software. The drug-disease targets were further subjected to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses by Metascape software. Additionally, the compound-target-pathway network was established in Cytosape to identify key compounds, targets, and pathways. Network pharmacology analysis screened out 96 active compounds and 837 potential targets within the 7 herbal/animal medicines of CHYS, among which 237 drug-disease common targets were identified. GO and KEGG analysis revealed the enrichment of fibrosis-related biological processes and pathways among the 237 common targets. Compound-target-pathway network analysis highlighted protein kinases Src and Akt1 as the top two targets associated with the anti-renal fibrosis effects of CHYS. In UUO mice, treatment with CHYS attenuates renal fibrosis, accompanied by suppressed expression and phosphorylation activation of Src. Unlike Src, CHYS reduced Akt1 phosphorylation without affecting its expression. In summary, network pharmacology and in vivo evidence suggest that CHYS exerts its anti-renal fibrosis effects, at least in part, by inhibiting the Src/Akt1 signaling axis.

Subscapular skinfold thickness, not other anthropometric and dual-energy X-ray absorptiometry-measured adiposity, is positively associated with the presence of age-related macular degeneration: a cross-sectional study from National Health and Nutrition Examination Survey 2005–2006

ObjectiveCurrent literature reveals an association between anthropometric measures of adiposity (AnthM) and age-related macular degeneration (AMD), but few have explored the disease association with imaging methods. This study aimed to...

The characteristics and influence of formula presenting in the 1950s

Formula presenting refers to the donation and submission of formula and prescriptions of Chinese medicine, organised by the Chinese government from 1954 to 1959. It was found that such donations and submissions of formula and prescriptions of Chinese medicine initiated from Sichuan, Anhui and Fujian due to their lack of medicine and physicians and the high incidence of epidemic diseases. This view is based on the literature such as books published at that time, local chronicles and relevant studies. The submissions were organised formally by the Ministry of Public Health in 1959 and distributed all over the country. It was found that the number of books published with such donated and submitted formula and prescriptions rose, and these books published were distributed in twenty-seven provinces, including country sides. It shows that such donation and submission of formula and prescriptions of Chinese medicine had a far-reaching impact on research and treatment for regional epidemic diseases and the development of traditional Chinese medicine.

Integrated metabolomics and transcriptomics analysis reveals the mechanism of Tangbi capsule for diabetic lower extremities arterial disease.

Tangbi capsule (TBC) is a traditional Chinese medicine prescription, which has the potential to improve the vascular insufficiency of lower extremities and limb numbness in diabetes. However, the potential mechanism remains unknown. This study aims to investigate the pharmacological effects and mechanism of TBC on rats with diabetic lower extremities arterial disease (LEAD). The mechanism of TBC on diabetic LEAD was investigated through metabolomics and transcriptomics analysis, and the main components of TBC were determined by mass spectrometry. The efficacy and mechanism of TBC on diabetic LEAD rats were investigated through in vitro experiments, histopathology, blood flow monitoring, western blot, and real-time polymerase chain reaction. Mass spectrometry analysis identified 31 active chemical components in TBC including (2R)-2,3-Dihydroxypropanoic acid, catechin, citric acid, miquelianin, carminic acid, salicylic acid, formononetin, etc. In vitro analysis showed that TBC could reduce endothelial cell apoptosis and promote angiogenesis. Histopathological analysis showed that TBC led to an obvious improvement in diabetic LEAD as it improved fibrous tissue proliferation and reduced arterial wall thickening. In addition, TBC could significantly increase the expression levels of HIF-1α, eNOS, and VEGFA proteins and genes while reducing that of calpain-1 and TGF-β, suggesting that TBC can repair vascular injury. Compared with the model group, there were 47 differentially expressed genes in the whole blood of TBC groups, with 25 genes upregulated and 22 downregulated. Eighty-seven altered metabolites were identified from the serum samples. Combining the changes in differentially expressed genes and metabolites, we found that TBC could regulate arginine biosynthesis, phenylalanine metabolism, pyrimidine metabolism, arachidonic acid metabolism, pyrimidine metabolism, arachidonic acid metabolism, nucleotide metabolism, vitamin B6 metabolism and other metabolic pathways related to angiogenesis, immune-inflammatory response, and cell growth to improve diabetic LEAD. TBC improved vascular endothelial injury, apoptosis, lipid accumulation, liver and kidney function, and restored blood flow in the lower extremities of diabetic LEAD rats. The mechanism of TBC in the treatment of diabetic LEAD may be related to the modulation of inflammatory immunity, lipid metabolism, and amino acid metabolism. This study presented preliminary evidence to guide the use of TBC as a therapy option for diabetic LEAD.

Online identification of chemical constituents in Mongolian medicine Zhachong-13 pills by UHPLC-Q-exactive Orbitrap MS

Zhachong-13 pills (ZC-13), as a traditional prescription of Mongolian medicine, are often used in the clinical practice of Mongolian hospitals for the treatment of stroke and rheumatic arthritis. In this experiment, UHPLC-Q-Exactive Orbitrap MS was used to explore the chemical composition of ZC-13. The results showed that 315 compounds were identified or inferred, including 56 alkaloids, 77 2-(2-phenylethyl)chromones, 61 flavonoids, 31 tannins, 8 coumarins, 16 lignans, 21 terpenoids, 5 amino acids, 19 organic acids, and 21 other components. In addition, the pharmacological activities related to anti-cerebral ischemia of these components were summarized. This result laid a foundation for further study on the pharmacodynamic material basis of ZC-13 and provided a scientific basis for the formulation of ZC-13 quality specifications.

Protocol for the development and validation of a Polypharmacy Assessment Score

BackgroundAn increasing number of people are using multiple medications each day, named polypharmacy. This is driven by an ageing population, increasing multimorbidity, and single disease-focussed guidelines. Medications carry obvious benefits, yet polypharmacy is also linked to adverse consequences including adverse drug events, drug-drug and drug-disease interactions, poor patient experience and wasted resources. Problematic polypharmacy is ‘the prescribing of multiple medicines inappropriately, or where the intended benefits are not realised’. Identifying people with problematic polypharmacy is complex, as multiple medicines can be suitable for people with several chronic conditions requiring more treatment. Hence, polypharmacy is often potentially problematic, rather than always inappropriate, dependent on clinical context and individual benefit vs risk. There is a need to improve how we identify and evaluate these patients by extending beyond simple counts of medicines to include individual factors and long-term conditions.AimTo produce a Polypharmacy Assessment Score to identify a population with unusual levels of prescribing who may be at risk of potentially problematic polypharmacy.MethodsAnalyses will be performed in three parts:1. A prediction model will be constructed using observed medications count as the dependent variable, with age, gender and long-term conditions as independent variables. A ‘Polypharmacy Assessment Score’ will then be constructed through calculating the differences between the observed and expected count of prescribed medications, thereby highlighting people that have unexpected levels of prescribing.Parts 2 and 3 will examine different aspects of validity of the Polypharmacy Assessment Score:2. To assess ‘construct validity’, cross-sectional analyses will evaluate high-risk prescribing within populations defined by a range of Polypharmacy Assessment Scores, using both explicit (STOPP/START criteria) and implicit (Medication Appropriateness Index) measures of inappropriate prescribing.3. To assess ‘predictive validity’, a retrospective cohort study will explore differences in clinical outcomes (adverse drug reactions, unplanned hospitalisation and all-cause mortality) between differing scores.DiscussionDeveloping a cross-cutting measure of polypharmacy may allow healthcare professionals to prioritise and risk stratify patients with polypharmacy using unusual levels of prescribing. This would be an improvement from current approaches of either using simple cutoffs or narrow prescribing criteria.

Dingxian pill alleviates hippocampal neuronal apoptosis in epileptic mice through TNF-α/TNFR1 signaling pathway inhibition

Ethnopharmacological relevanceDingxian Pill (DXP), a famous traditional Chinese medicine prescription, and has been widely proven to have positive therapeutic effects on “Xianzheng” (the name of epilepsy in ancient China). However, the anti-epileptic molecular mechanisms of DXP are not yet fully understood and remain to be further investigated. Aim of the studyTo elucidate the molecular mechanism of DXP's improvement in epileptic neuronal loss, damage and apoptosis by regulating TNF-α/TNFR1 signaling pathway. Materials and methodsSixty Kunming mice were randomly divided in 6 groups: control group (equal volume of normal saline), model group (180 mg kg−1 pilocarpine hydrochloride – used to establish the epilepsy animal model), carbamazepine group (30 mg kg−1), and low, medium, and high-dose Dingxian Pill groups (4.08, 8.16, and 16.32 g kg−1, respectively – oral administration once daily for 2 weeks). Successful establishment of the epileptic mouse model was monitored with electroencephalography. Pathological changes in hippocampal tissue were analyzed with hematoxylin-eosin staining. Hippocampal neuronal apoptosis was analyzed with TUNEL staining. TNF-α, TNFR1, TRADD, FADD, and caspase-8 mRNA and protein expression levels in hippocampal tissue were analyzed with real-time quantitative polymerase chain reaction, immunohistochemistry, and Western blot, respectively. Cleaved caspase-8 protein levels in hippocampal tissue were measured with immunohistochemistry and Western blot. ResultsCompared to control, the model group showed an increase in continuous epileptic discharge waves on EEG, a damaged hippocampal neuron morphological structure, increased hippocampal neuronal apoptosis, and significantly increased TNF-α, TNFR1, TRADD, FADD, and caspase-8 mRNA and protein levels, and increased caspase-8 cleavage (P < 0.05). Compared to the model group, the carbamazepine group as well as the low-, medium-, and high-dose Dingxian Pill groups showed decreased epileptic discharges on EEG, an obvious hippocampal neuron morphological structure restoration, varying degrees of attenuated hippocampal neuronal apoptosis, and significantly decreased TNF-α, TNFR1, TRADD, FADD, and caspase-8 mRNA and protein levels as well as decreased caspase-8 cleavage (P < 0.05). ConclusionsDingxian Pill exerts an anti-epileptic effect through inhibition of TNF-α/TNFR1 signaling pathway-mediated apoptosis in hippocampal neurons.

Integrating UHPLC-Q-TOF-MS/MS, network pharmacology, bioinformatics and experimental validation to uncover the anti-cancer mechanisms of TiaoPi AnChang decoction in colorectal cancer

Ethnopharmacological relevanceThe TiaoPi AnChang Decoction (TPACD), a Traditional Chinese Medicine (TCM) prescription based on Xiangsha Liujunzi Decoction, has demonstrated clinical efficacy as an adjuvant therapy for colorectal cancer (CRC) patients. However, its specific ingredients and potential mechanisms of action remain unclear. Aim of the studyTo identify the primary active ingredients of TPACD, their molecular targets, and potential mechanisms underlying the efficacy of TPACD in CRC treatment. Materials and methodsThis study investigated the clinically validated TCM formula TPACD. In vitro and in vivo experiments were used to demonstrate TPACD's regulatory effects on various malignant phenotypes of tumors, providing basic research support for its anti-cancer activity. To understand its pharmacodynamic basis, we utilized ultra-high performance liquid chromatography-quadrupole-time-of-flight-mass spectrometry/mass spectrometry (UHPLC-Q-TOF-MS/MS) to analyze TPACD constituents present in the bloodstream. Network pharmacology and bioinformatics analyses were used to identify potential active components and their molecular targets for TPACD's therapeutic effects in CRC. Subsequent experiments further elucidated its pharmacological mechanism. ResultsTPACD inhibits various malignant cellular processes, such as cell proliferation, apoptosis, migration, and invasion, and has shown potential anti-CRC activities both in vitro and in vivo. Following the identification of 109 constituents absorbed into the blood from TPACD, network pharmacology analysis predicted 42 potential anti-CRC targets. Clinical analyses highlighted three genes as prognostic key genes of TPACD's therapeutic action: C-X-C motif chemokine ligand 8 (CXCL8), fatty acid binding protein 4 (FABP4), and matrix metallopeptidase 3 (MMP3). Drug sensitivity analyses, molecular docking simulations and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) identified MMP3 as the most promising target for TPACD's anti-CRC action. Enzyme activity assays confirmed that TPACD inhibits MMP3 enzyme activity. Surface plasmon resonance (SPR) characterized the binding affinity between MMP3 and effective active components of TPACD, including luteolin, quercetin, kaempferol, and liensinine. ConclusionsTPACD exhibits anti-CRC activity in vitro and in vivo, with MMP3 identified as a critical target. The active compounds, including luteolin, quercetin, kaempferol, and liensinine, absorbed into the bloodstream, contribute to TPACD's efficacy by targeting MMP3.

A multi-component, multi-target, and multi-pathway prediction method for Chinese medicine based on the combination of mass spectrometry analysis and network analysis: An example using Weifuchun

Weifuchun, a Chinese medicinal prescription made from herbs of natural origin including Hongshen (Ginseng Radix et Rhizoma Rubra), Xiangchacai (Rabdosia Amethystoides), and Zhiqiao (Aurantii Fructus), has attracted increasing attention for clinically treating chronic atrophic gastritis, which is characterized by the chronic inflammation of the gastric mucosa leading to progressive loss of gastric glandular cells. To investigate the active ingredients and potential mechanisms of WFC, it was analyzed using a novel multi-component, multi-target, and multi-pathway prediction method. High/ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (HPLC/UPLC-Q-TOF-MS) was employed to separate and profile the chemical constituents of WFC with high precision and efficiency. Network analysis and molecular docking were used to predict bioactive compounds and their interactions with biological targets. The results highlight 42 significant compounds potentially contributing to the therapeutic effects of WFC by effecting on several key pathways, including proved PI3K/Akt, NF-κB, and JAK/STAT signaling pathways. This study showcases the efficacy of combining advanced chromatographic techniques with computational methods to elucidate the pharmacological mechanisms of complex botanical drugs.

Shengqing Jiangzhuo capsule ameliorates diabetic nephropathy by improving Keap1/Nrf2 signaling pathway.

Diabetic nephropathy (DN) is a major contributor to end-stage renal failure, and lacking effective treatment options. Shengqing Jiangzhuo capsule (SQJZJN), a traditional Chinese medicine prescription with known efficacy in chronic kidney disease, has not been thoroughly investigated for its potential in DN protection. Eight-week-old male C57BLKS/J db/db, C57BLKS/J db/m mice, and human glomerular mesangial cell (HMC) cells cultured with high glucose were used as experimental models in this study. The in vivo investigation showed that SQJZJN can significantly ameliorate renal pathological damage, reduce serum creatinine, and lower urinary microalbumin levels in db/db mice. In vitro, SQJZJN treatment mitigated advanced glycation end products (AGEs) and reactive oxygen species (ROS), leading to a reduction in renal cell apoptosis. Mechanistically, SQJZJN activated the Keap1/Nrf2/ARE pathway by promoting nuclear factor erythroid-derived 2-related factor 2 (Nrf2), γ-glutamylcysteine synthetase heavy subunit (γ-GCS), and Heme oxygenase-1 (HO-1) expressions, while decreasing Kelch-like ECH-associated protein 1 (KEAP1) expressions. These findings suggest that SQJZJN exerts a protective effect on DN, potentially through the activation of the Keap1/Nrf2/ARE pathway.

A Systematic Review of Antibiotic Use in Humans in Nigeria and Its Potential Contribution to Rising Antimicrobial Resistance

INTRODUCTION: The value of medicines is lost when these resources are not used rationally. Out of 12 developing countries, Nigeria has the third highest percentage of antibiotic prescriptions, at 48%. Antibiotic misuse results in limited efficacy, which can lead to the emergence of antimicrobial resistance. We conducted a systematic review to synthesise the evidence on antibiotic use in humans in Nigeria. METHODS: We conducted a systematic review of medicine use behaviour by patients and prescription behaviour by health workers, which were searched for in articles published in English from 2000 to 2017. Data was entered into purpose-built templates. Key quantitative indicators were extracted and summarised as frequencies and proportions, while free-text responses were synthesised. RESULTS: The systematic review determined that the median prevalence of persons using antibiotics without prescription is 46.7%. The drivers of irrational antibiotic use included poor regulation of medicines and premises, a chaotic medicine distribution system, limited licensed medicine prescribers, over-the-counter (OTC) sales of antibiotics, patients’ demand for antibiotics, and access to health insurance. DISCUSSION: Irrational antibiotic use is widespread in humans and animals. We recommend that the government enforce regulations on antibiotic sales to humans and animals and increase awareness of irrational antibiotic use and AMR in Nigerian communities through a whole-of-society approach. The collated information was used to develop a National Action Plan on AMR in 2017.

Mechanism of action of the Banxia–Xiakucao herb pair in sleep deprivation: New comprehensive evidence from network pharmacology, transcriptomics and molecular biology experiments

Chinese herb pairs are the most basic and compressed examples of Chinese herbal combinations and can be used to effectively explain the fundamental concepts of traditional Chinese medicine prescriptions. These pairings have gained significant interest due to their subtle therapeutic benefits, minimal side effects, and efficacy in treating complicated chronic conditions. The Banxia-Xiakucao Chinese herb pair (BXHP) consists of Pinellia ternata (Thunb.) Breit. (Banxia) and Prunella vulgaris L. (Xiakucao). This formula was documented in The Medical Classic of the Yellow Emperor approximately 2000 years ago，and clinical research has demonstrated that BXHP effectively treats insomnia. This study aimed to evaluate the efficacy and therapeutic mechanism of the BXHP through a comprehensive strategy involving network pharmacology, molecular docking, transcriptomics, and molecular biology experimental validation. The composition of BXHP was characterized using the UPLC-Q-TOF-MS. The active compounds were screened to find drug-likeness compounds by analyzing the ADME data. To predict the molecular mechanism of BXHP in sleep deprivation (SD) by network pharmacology and molecular docking. We established a rat model of SD and the in vivo efficacy of BXHP was verified through the pentobarbital sodium righting reflex test, behavioral assays, enzyme-linked immunosorbent assay, transmission electron microscopy, HE staining, and Nissl staining, and the underlying molecular mechanism of BXHP in SD was revealed through transcriptomic and bioinformatic analyses in conjunction with quantitative real-time PCR, Western blot, and immunofluorescence staining. In the present study, we showed for the first time that BXHP reduced sleep latency, prolongs sleep duration, and improves anxiety; lowered serum CORT, IL6, TNF-α and MDA levels; decreased hypothalamic Glu levels; and elevated hypothalamic GABA and 5-HT levels in SD rats. We found 16 active compounds that acted on 583 targets, 145 of which are related to SD. By modularly dissecting the PPI network, we discovered three critical targets, Akt1, CREB1, and PRKACA, all of which play important roles in the effects of BXHP on SD. Molecular docking resulted in the identification of 16 active compounds that strongly bind to key targets. The results of GO and KEGG enrichment analyses of network pharmacology and transcriptomics focused on both the regulation of circadian rhythm and the cAMP signaling pathway, which strongly demonstrated that BXHP affects SD via the cAMP-PKA-CREB-Circadian rhythm pathway. Molecular biology experiments verified this hypothesis. Following BXHP administration, PKA and CREB phosphorylation levels were elevated in SD rats, the cAMP-PKA-CREB signaling pathway was activated, the expression levels of the biological clock genes CLOCK, p-BMAL1/BMAL1, and PER3 were increased, and the rhythmicity of the biological clock was improved. The active compounds in BXHP can activate the cAMP-PKA-CREB-Circadian rhythm pathway, improve the rhythmicity of the biological clock, promote sleep and ameliorate anxiety, which suggests that BXHP improves SD through a multicomponent, multitarget, multipathway mechanism. This study is important for the development of herbal medicines and clinical therapies for improving sleep deprivation.

Characterization of active alkaloids and metabolites in rats after oral administration of Zuojin Pill using UHPLC-Q-TOF-MS combined with bioinformatics and molecular docking analyses

Zuojin Pill (ZJP), a traditional Chinese medicine prescription composed of Rhizoma Coptidis and Euodiae Fructus in the ratio of 6:1 (w/w), has been widely used for the treatment of gastric disorders. However, an in-depth understanding of in vivo metabolism and distribution profiles of protoberberine alkaloids (PBAs) and indole alkaloids (IDAs) in ZJP is lacking. In this study, a method using ultra-high performance liquid chromatography coupled with quadruple time-of-flight mass spectrometry (UHPLC-Q-TOF-MS) was developed to systematically screen the alkaloids and their metabolites in rat plasma and various tissues after oral administration of ZJP. Furthermore, bioinformatics and molecular docking analyses were conducted to elucidate the contribution of the alkaloids and metabolites enriched in the stomach to the therapeutic effect of ZJP on gastritis. A total of 33 compounds, including 7 prototype alkaloids and 26 metabolites, were chemically defined or tentatively identified in this work. The metabolic pathways of PBAs (hydroxylation, oxidation, reduction, demethylation, demethylenation, glucuronide conjugation, sulfate conjugation) and IDAs (hydroxylation, glucuronide conjugation) were revealed. Notably, 7 prototype alkaloids and 18 metabolites were detected in the stomach, indicating their propensity for gastric distribution. These alkaloids and metabolites showed strong affinities with the 7 hub targets associated with gastritis, such as CCR7, CXCR4, IL6, IFNG, CCL2, TNF, and PTPRC, and could be considered the potential active substances of ZJP for treating gastritis. In conclusion, this study clarified the gastric distribution propensity of PBAs and IDAs and their metabolites, as well as their favorable binding interactions with gastritis-related targets, which could provide essential data for the further study of the pharmacodynamic material basis and gastroprotective mechanism of ZJP.

Sangju Cold Granule exerts anti-viral and anti-inflammatory activities against influenza A virus in vitro and in vivo

Ethnopharmacological relevanceSangju Cold Granule (SJCG) is a classical traditional Chinese medicine (TCM) prescription described in “Item Differentiation of Warm Febrile Diseases”. Historically, SJCG was employed to treat respiratory illnesses. Despite its popular usage, the alleviating effect of SJCG on influenza A virus infection and its mechanisms have not been fully elucidated. Aim of the studyInfluenza is a severe respiratory disease that threatens human health. This study aims to assess the therapeutic potential of SJCG and the possible molecular mechanism underlying its activity against influenza A virus in vitro and in vivo. Materials and methodsUltrahigh-performance liquid chromatography (UPLC)-Q-Exactive was used to identify the components of SJCG. The 50% cytotoxic concentration of SJCG in MDCK and A549 cells were determined using the CCK-8 assay. The activity of SJCG against influenza A virus H1N1 was evaluated in vitro using plaque reduction and progeny virus titer reduction assays. RT-qPCR was performed to obtain the expression levels of inflammatory mediators and the transcriptional regulation of RIG-I and MDA5 in H1N1-infected A549 cells. Then, the mechanism of SJCG effect on viral replication and inflammation was further explored by measuring the expressions of proteins of the RIG-I/NF-kB/IFN(I/III) signaling pathway by Western blot. The impact of SJCG was explored in vivo in an intranasally H1N1-infected BALB/c mouse pneumonia model treated with varying doses of SJCG. The protective role of SJCG in this model was evaluated by survival, body weight monitoring, lung viral titers, lung index, lung histological changes, lung inflammatory mediators, and peripheral blood leukocyte count. ResultsThe main SJCG chemical constituents were flavonoids, carbohydrates and glycosides, amino acids, peptides, and derivatives, organic acids and derivatives, alkaloids, fatty acyls, and terpenes. The CC50 of SJCG were 24.43 mg/mL on MDCK cells and 20.54 mg/mL on A549 cells, respectively. In vitro, SJCG significantly inhibited H1N1 replication and reduced the production of TNF-α, IFN-β, IL-6, IL-8, IL-13, IP-10, RANTES, TRAIL, and SOCS1 in infected A549 cells. Intracellularly, SJCG reduced the expression of RIG-I, MDA5, P–NF-κB P65 (P–P65), P-IκBα, P-STAT1, P-STAT2, and IRF9. In vivo, SJCG enhanced the survival rate and decreased body weight loss in H1N1-infected mice. Mice with H1N1-induced pneumonia treated with SJCG showed a lower lung viral load and lung index than untreated mice. SJCG effectively alleviated lung damage and reduced the levels of TNF-α, IFN-β, IL-6, IP-10, RANTES, and SOCS1 in lung tissue. Moreover, SJCG significantly ameliorated H1N1-induced leukocyte changes in peripheral blood. ConclusionsSJCG significantly reduced influenza A virus and virus-mediated inflammation through inhibiting the RIG-I/NF-kB/IFN(I/III) signaling pathway. Thus, SJCG could provide an effective TCM for influenza treatment.

The signaling pathways of selected traditional Chinese medicine prescriptions and their metabolites in the treatment of diabetic cardiomyopathy: a review.

Diabetic cardiomyopathy (DCM) is a myocardial-specific microvascular disease caused by diabetes that affects the structure and function of the heart and is considered to be the leading cause of morbidity and death in patients with diabetes. Currently, there is no specific treatment or preventive drug for DCM, and there is an urgent need to develop new drugs to treat DCM. Traditional Chinese medicine (TCM) has rich experience in the treatment of DCM, and its characteristics of multi-target, multi-pathway, multi-component, and few side effects can effectively deal with the complexity and long-term nature of DCM. Growing evidence suggests that myocardial fibrosis, inflammation, oxidative stress, apoptosis, cardiac hypertrophy, and advanced glycation end product deposition were the main pathologic mechanisms of DCM. According to the pathological mechanism of DCM, this study revealed the potential of metabolites and prescriptions in TCM against DCM from the perspective of signaling pathways. The results showed that TGF-β/Smad, NF-κB, PI3K/AKT, Nrf2, AMPK, NLRP3, and Wnt/β-catenin signaling pathways were the key signaling pathways for TCM treatment of DCM. The aim of this study was to summarize and update the signaling pathways for TCM treatment of DCM, to screen potential targets for drug candidates against DCM, and to provide new ideas and more experimental evidence for the clinical use of TCM treatment of DCM.

O-315 Familial endocrine disease increases the risk of pregnancy loss and recurrent pregnancy loss– a nationwide register-based study of 366,548 Danish women

Abstract Study question Is endocrine disease and pregnancy loss associated, and can the association be due to inherited genetic causes? Summary answer Endocrine disease is associated with an increasing number of pregnancy losses. Endocrine disease in parents and siblings increases a woman’s risk of pregnancy loss. What is known already One in four pregnancies results in a pregnancy loss, but only 60% hereof are due to detectable chromosome abnormalities. The remaining 40% are suspected to be caused by other factors such as maternal comorbidity. Recurrent pregnancy loss (RPL) is defined as 2 or 3 consecutive losses and affects 1-3% of couples trying to conceive. Pregnancy loss has been associated with endocrine diseases, e.g., thyroid disease, diabetes, or PCOS. Furthermore, thyroid autoimmunity has been associated with RPL. It is unknown if other endocrine diseases are associated with RPL. To date, there is no evidence for a genetic basis of the risk. Study design, size, duration Danish nationwide cohort study including data from the Danish National Health Registers on ICD-8/ICD-10 diagnosis codes, all pregnancy outcomes and redeemed medicine prescriptions. Endocrine disease was defined as any endocrine disease (e.g., diabetes, thyroid disease, PCOS etc.). The study included in total 366,548 women, including 54,394 women with endocrine disease and 312,154 without endocrine diseases. The study included data from 1973 to 2022. Linkage to parent and siblings was available through the Multi-Generation Registry Lite. Participants/materials, setting, methods The cohort consisted of women born between 1977-1993 with ≥1 pregnancy. The exposure was endocrine disease. Logistic regression models adjusted for birth year provided odds ratios (ORs) for endocrine diseases according to number of pregnancy losses. Parent-offspring and sibling regression provided estimates for familial aggregation, assessing the degree of resemblance between relatives to infer the potential influence of shared familial factors. The variance-covariance matrix was adjusted using the Huber-White method to account for familial clustering. Main results and the role of chance We found a significant association between endocrine disease and pregnancy loss increasing with the number of pregnancy losses; OR and 95%CI for one loss 1.15 (1.12-1.17), two losses 1.31 (1.24-1.38) and ≥ three losses 1.81 (1.70-1.93). Further, endocrine disease was associated with RPL (OR 1.87, (1.74-2.00)). The association was strongest with primary RPL (OR 2.13 (1.94-2.35)), compared to secondary RPL (OR 1.58 (1.42-1.76)). In cases where either of a woman’s parents was diagnosed with an endocrine disease, she also had a higher risk of pregnancy loss (OR 1.06 (1.04–1.08)) and RPL (OR 1.07 (1.01–1.14)), compared to a woman with parents without endocrine disease. If a woman’s sibling had an endocrine disease, the OR for pregnancy loss was 1.07 (1.03–1.11) and RPL 1.17 (1.03–1.33). Further stratification into individual endocrine phenotypes revealed that type 2 diabetes conferred a comparable risk: for offspring of parents with this condition, the risk of pregnancy loss increased to an OR of 1.08 (1.06–1.11), and for siblings, the OR was 1.13 (1.02–1.25). This effect persisted even when adjusting for the disease in the individual experiencing the loss. Limitations, reasons for caution Very early pregnancy losses are often not handled at the hospital and therefore not part of the Danish registers. Thus, there is a risk of underdiagnosed pregnancy losses, although this would be the case for both the exposed and control cohort. Wider implications of the findings Women with RPL should be investigated for endocrine disease, and family history of endocrine disease is an important and novel risk factor that should guide the diagnostic work-up. Identifying the shared mechanism or pleiotropic effects between pregnancy loss and endocrine disease should be a focus area. Trial registration number Research related to this project was supported by the Novo Nordisk Foundation (ID: NNF22OC0077221, NNF23OC0087269) and the William Demant Foundation (salary for Pia Egerup).

Effects of Extraction Methods on Volatile Oil Profiles of Cinnamomi ramulus–Zingiberis rhizoma recens Couplet Medicines

Cinnamomi ramulus (CR, Guizhi in Chinese) and Zingiberis rhizoma recens (ZRR, Shengjiang in Chinese) are couplet medicines (drug pairs or drug combinations) in traditional Chinese medicine prescriptions. They contain a high amount of volatile oils which endow them with unique flavors and pharmacological activities. Extraction methods have a great influence on the extraction efficiency and composition of volatile oils. Firstly, the volatile oils of CR and ZRR were extracted by steam distillation (SD) and analyzed by GC-MS to obtain their chemical profiles. In total, 35 and 55 compounds were identified in the volatile oils of CR and ZRR, respectively. In order to find a suitable extraction method for the couplet medicine CR-ZRR, subsequently, steam distillation (SD), azeotropic distillation (AD) and supercritical fluid extraction (SFE) were applied to extract the volatile oils from CR-ZRR. The average extraction yields by SD, AD and SFE were 0.573%, 0.62% and 2.135%, respectively. The chemical composition of the volatile oils was then analyzed by GC-MS. In total, 73, 59 and 71 compounds were identified from the extracts obtained by SD, AD and SFE, respectively. Principal component analysis (PCA) and OPLS-DA showed that citral, (E)-cinnamaldehyde dimethyl acetal, zingiberene, cinnamaldehyde and β-sesquiphellandrene were the main contributors to distinguish the volatile oils that were obtained by different processes. Considering the chemical diversity and the total content of the main bioactive components of the volatiles oils, SD was more suitable for CR-ZRR volatile oil extraction. This study provides a basis for elucidating the chemical composition and suitable extraction method for the volatile oils of CR-ZRR.

Rujin Jiedu decoction protects against influenza virus infection by modulating gut microbiota

BackgroundRujin Jiedu decoction (RJJDD) is a classical prescription of Traditional Chinese Medicine that has long been applied to treat pneumonia caused by external infection, but whether and how it benefits influenza virus therapy remains largely unclear. The aim of this study was to investigate the anti-inflammatory effect of RJJDD on the mouse model of influenza and to explore its potential mechanism. MethodsThe mice were mock-infected with PBS or infected with PR8 virus followed by treatment with RJJDD or antiviral oseltamivir. The weight loss and morbidity of mice were monitored daily. Network pharmacology is used to explore the potential pathways that RJJDD may modulate. qRT-PCR and ELISA were performed to assess the expression of inflammatory cytokines in the lung tissue and macrophages. The intestinal feces were collected for 16S rDNA sequencing to assess the changes in gut microbiota. ResultsWe demonstrate that RJJDD protects against IAV-induced pneumonia. Comprehensive network pharmacology analyses of the Mass Spec-identified components of RJJDD suggest that RJJDD may act through down-regulating key signaling pathways producing inflammatory cytokines, which was experimentally confirmed by cytokine expression analysis in IAV-infected mouse lung tissues and IAV single-strand RNA mimic R837-induced macrophages. Furthermore, gut microbiota analysis indicates that RJJDD prevented IAV-induced dysbiosis of host intestinal flora, thereby offering a mechanistic explanation for RJJDD's efficacy in influenza pneumonia. ConclusionThis study defines a previously uncharacterized role for RJJDD in protecting against influenza likely by maintaining homeostasis of gut microbiota, and provides a new therapeutic option for severe influenza.