Snack bars, known as energy bars, are widely consumed worldwide as highly nutritive on-the-go products. Due to the date fruit’s significant nutritional and functional characteristics, it can be an exceptional choice for developing snack bars. Dates contain a wide range of macro- and micronutrients known for their strong bioactive properties. The functional properties of date fruit have been demonstrated in the literature and include antioxidant, anti-inflammatory, anti-tumor, antihypertensive, and antimicrobial activities. This review summarizes the available studies investigating the potential application of dates for developing nutritive and functional snack bars. Date paste was used as a main ingredient at 55–90% concentrations. In addition, protein sources were used to provide protein-rich snack bars, as date fruit is considered high in carbohydrates and low in protein. Skim milk powder was the most common and favorable protein source, delivering significant amounts of protein with limited negative effects on sensory attributes. Incorporating other ingredients, such as cereals or legumes, was also explored; adding such dry ingredients can promote positive nutritional effects along with improving sensory attributes, mainly in terms of the bars’ textures. Dry ingredients can significantly lower moisture content, reducing the bars’ fracturability to acceptable ranges. Reduced moisture content can also significantly enhance the shelf-life stability, as observed by limited microbial growth. Furthermore, the incorporation of bioactive or functional ingredients such as fruit peels, plant seeds, or plant leaf extracts was also reported; such ingredients promoted significant enhancements in the contents of phenolics or flavonoids, for instance, leading to an increase in the bars’ antioxidant potential. Though dates are rich in such bioactive components, incorporating additional bioactive ingredients can boost the dates’ functional properties. In conclusion, this review shows the high potential of the application of dates for developing nutritious and functional snack bars. Taking this into account, the snack bar market has grown remarkably over the past decade; thus, providing well-balanced, nutritious, and functional date-based bars in markets worldwide is expected to show positive consumer acceptance. The demand for easy and immediate nutrient-dense meal replacement has been growing for the past two decades with growing concerns about health and fitness. Commercial bar-shaped snack products, known as snack or energy bars, are an excellent choice and the most common on-the-go products serving consumers’ health demands. Snack bars are mostly manufactured to provide good amounts of nutrients and serve as energy boosters, thus gaining the common term “energy bars”. Snack bars can be enjoyable, suitable, and equally nutritious for all age groups but are mostly consumed by highly active individuals such as athletes ,2. The gDue to its rich nutritional composition, the date palm fruit can be an exceptional choice for manufacturing snack bars. Although they are considered a rich source of carbohydrates, dates can provide significant amounts of good-quality nutrients, such as dietary fiber , unsaturated fatty acids , and a wide range of micronutrients . Dates are also considered low in protein, though studies have found significant essential amino acids in dates, such as lysine and histidine, that are lacking in most fruits. In addition, in dates, various types of bioactive phytochemicals, such as phenolic acids, polyphenols, and carotenoids, were found in significant amounts ,6,7,8. SPhoenix dactylifera”, “date bars”, “energy bars”, “snack bars”, “date energy bars”, and “date snack bars”.Apart from the substantial nutritional aspects of dates, from an economic point of view, dates are considered highly valuable. Their cultivation has shown a significant growth trend over the past three decades. Date production is widespread worldwide, mainly in Middle Eastern and Northern African countries, which share more than 70% of world production. Countries such as Egypt and Saudi Arabia are the top countries producing dates and showed a growth trend of 222 and 197%, respectively, from 1990 to 2021 [P. dactylifera) is one of the most important crops of the Arecaceae family; it has been grown since ancient times, more than 4000 years ago, mainly in Middle Eastern and Northern African countries, predominantly in dry and arid regions. It has been cultivated primarily for cultural, nutritional, environmental, religious, and social development purposes [Date fruit  [−1 [−1 [Dates are considered a relatively good source of different vitamins , mainly ectively . Other sectively . Date pi variety . Some daety [−1) ,28. Vita[−1) [−1 . Vitamin [−1 [−1 . However [−1 [−1 .Minerals such as potassium, calcium, and magnesium are found in relatively high amounts in dates, ranging from 0.05 to 0.9%. The most abundant element found in dates is mostly potassium, followed by calcium. Some date varieties, such as Sukkari, contained higher mineral amounts than other fruits, such as pomegranate or mango ; others −1  per 100 g−1 . High amMalaysia . Some fl−1 while fresh, but after drying, the content increased to 339 mg FAE 100 g−1 [The total phenolic content can significantly vary between fresh and dried dates; the Khalas variety, for instance, contained 134 mg ferulic acid equivalents (FAE) per 100 g 100 g−1 . Phenoli 100 g−1 .−1 for total flavonoid compounds [Several detected phenolic compounds were found in various date varieties; different phenolic acids were detected in Ajwa dates, including ferulic, hydroxybenzoic, gallic, caffeic, vanillic, chlorogenic, isovanillic, chlorogenic, protocatechuic, isoferulic, and hydroxycinnamic acids . Other dompounds . Such a ompounds . Others ompounds . Dates aompounds .−1 of body weight (BW) were significantly effective in improving the levels of antioxidant markers, including glutathione transferase, catalase, glutathione reductase, and malonaldehyde [Elevating the body’s antioxidant system, or in other words, eliminating oxidative stress, a state caused by the excess production of free radicals leading to the development and progression of various diseases, can be achieved by consuming dates. The rich contents of different date phytochemicals were significantly correlated in some studies with dates’ antioxidant potential ,24. For aldehyde ,41,42,43aldehyde .−1 BW of date pit powder to carbon tetrachloride (CCI4)-induced rats for two weeks resulted in significant improvements in the levels of some pro-inflammatory cytokines, such as tumor necrosis factor (TNF-α) and interferon-gamma (IFN-γ) [Besides elevating the antioxidant system, date intake can also reduce the body’s inflammatory state, another critical system in the development and progression of various diseases. A reduction in the inflammatory state is also attributed to dates’ strong antioxidant potential; inflammatory states are mostly initiated due to oxidative stress due to free radicals . The adm (IFN-γ) . A syste (IFN-γ) . Other s (IFN-γ) ,47,48,49−1 BW to animal models was effective in alleviating postprandial glycemia. These positive effects were attributed to inhibitory activities against some enzymes related to type 2 diabetes, such as α-glucosidase, an intestinal enzyme responsible for regulating glucose availability for intestinal absorption [−1 BW resulted in antidiabetic effects based on improvements in glucose uptake stimulation, cellular glycogen synthesis, and/or pancreatic cell protection against dexamethasone injection [Despite the rich contents of sugars in dates, some in vivo studies have reported hypoglycemic effects after the intake of dates; El Abed et al.  showed tsorption . In a resorption . Anothernjection . Date innjection . Howevernjection . In addinjection . TherefoThough date fruit is generally considered low in glycemic load, a study examining the glycemic index (GI) of five different date varieties in healthy individuals showed that the intake of 50 g of dates with available carbohydrates equivalent to 50 g of glucose had an average GI ranging from 30 to 69 . In anot−1 BW were orally administered to hypercholesterolemic rabbits for 10 weeks. Supplementation with date extracts led to enhancements in lipid metabolism based on reduced levels of total cholesterol, low-density lipoprotein (LDL), and triglycerides. In contrast, the high-density lipoprotein (HDL) level was increased compared to the control group [−1 BW of a date suspension for 14 days led to reductions in the levels of total cholesterol, triglycerides, and LDL, while levels of HDL showed an increase, indicating the anti-hypercholesterolemic effects of dates [−1 BW for 21 days, the results showed significant dose-dependent enhancements in lipid metabolism based on reduced levels of total cholesterol and LDL as well as the atherogenic index [Due to their rich dietary fiber content and various bioactive phytochemicals, dates were also reported to promote cholesterol-lowering activities. Date extracts at concentrations ranging from 125 to 1000 mg kgol group . In anotof dates . Date piic index . The choic index .−1 BW for 11 days synergically with an injection of doxorubicin [−1 BW promoted anti-tumor activities mediated by cell proliferation inhibition. It was shown that the Barhi date extract exerts inhibitory effects on critical signaling pathways related to cellular growth and functions, such as PTEN and AKT, namely, phosphatase and tensin homolog and protein kinase B, respectively. Date leaf extracts were also reported to promote significantly stronger anti-tumor activities than date pulp or pit extracts; the results showed that the phenolic contents and antioxidant capacity of the date leaf extract were significantly higher than those of the pulp or pit [Discovering effective anti-tumor agents is of major interest in the research field due to the numerous adverse effects of existing cancer treatments, such as radiotherapy and chemotherapy. The functional properties derived from fruits’ phytochemicals, such as flavonoids and other phenolic compounds, have been linked to anti-tumor activities ,65. Dateorubicin , a frequorubicin . The intorubicin . In anotorubicin , the daip or pit . Consistp or pit ,16,71,72Various pharmacological effects of date fruit were also reported in several studies. The high potassium content in dates can significantly benefit hypertensive individuals; the role of potassium in regulating blood pressure is well established . Dates aBacillus subtilis, Salmonella typhi, Enterococcus faecalis, Escherichia coli, Bacillus cereus, and Staphylococcus aureus [Fusarium oxysporum were also reported after using date extract, indicating the potential usage of dates against food spoilage microbes [Pseudomonas phage at a relatively extremely low concentration of less than 10 mg ml−1; such inhibitory effects were likely attributable to the date pit extract’s ability to bind directly to phages [Herpesviridae family known to cause a range of partly life-threatening infections in humans and animals and considered a significant public health concern. The ethanolic date extract at a 5% concentration showed more effective and faster anti-viral activities than other concentrations compared to acyclovir, a reference drug used for treating HSV-1 infections [Moreover, dates were reported to promote antimicrobial effects; different date varieties’ extracts were found in vitro and in vivo to exert inhibitory effects against Gram-positive and Gram-negative bacteria, such as s aureus . Inhibitmicrobes . Furthero phages . In anotfections . HoweverThe application of different types of fruits for developing snack bars is widely used; fruit-based bars have been developed to provide excellent snack bars containing natural sugars, vitamins, and minerals with other bio-nutritive components to meet consumers’ required daily nutritional intake . Date frDepending on the manufacturing target, date bars can be formulated with only one or two ingredients or with many several ingredients. Regarding protein enrichment, many different protein sources can be used to prepare date bars, including either dairy or plant sources. Proteins include milk powders or its products , soy proteins, or other plant-derived proteins . Many other ingredients can also be incorporated to deliver nutritionally balanced, functional, flavorful, and enjoyable date bars, including cereals , legumes , dried fruits  and nuts  with the addition of small amounts of spices for seasoning . These different ingredients can deliver functional attributes; other novel ingredients can also deliver much stronger functional attributes, such as plant seeds and fruit polyphenol extracts ,82,83,84−1. This formulation of date bars also had excellent amounts of minerals and vitamins. Increases in calcium, iron, phosphorus, and magnesium of 30, 26, 150–250, and 13–35%, respectively, were observed compared to control bars. High phosphorus and magnesium contents were mostly derived from single-cell protein, while calcium was from dry skim milk. Single-cell protein also contains good amounts of vitamins such as thiamin and riboflavin, which were significantly increased in the bars. However, after 6 months of storage at ambient temperature (25–37 °C), there was a loss in both vitamins of 10–25%, while storage at 5 °C maintained both vitamins’ contents, indicating the importance of the storage time and temperature in maintaining their contents. These two factors can also affect the bars’ sensory quality; bars stored at 5 °C had higher scores for flavor and texture compared to those stored at higher temperatures. However, storage at ambient temperature for 3–6 months led to significant improvements in the overall sensory quality of bars made from dates alone (control bars) compared to their initial quality, while enriched bars stored under the same conditions showed the opposite effect.Several researchers worldwide have prepared multiple variations of date-based bars ; this seYeast-derived proteins such as Toruway and Zyest were used in another study in the 1980s. In a similar approach to that used by Kamel and Kramer , Sawaya Moreover, soy protein isolates and skim milk powder contain higher amounts of minerals than dates. Major increases (2–4 times) in several minerals, such as sodium, potassium, and calcium, were observed in fortified bars. The addition of these ingredients caused nonsignificant changes in overall sensory acceptability compared to control bars. Similar to Kamel and Kramer’s results , storage−1 for Sukkari bars vs. Nabtat-Ali bars, respectively). However, raw dates showed higher antioxidant activity than date bars; exposure to higher temperatures during the preparation processes applied in this study, such as heating or boiling, could significantly reduce the total phenolics. Bars’ sensory quality can also differ based on the variety used; Sukkari bars were tenderer or softer than Nabtat-Ali bars, and the latter required twice the time for chewing. Sukkari dates also contributed to a better color. However, neither bar received higher scores in flavor or overall acceptability, indicating that additional work, such as adding suitable flavoring agents, is needed.However, these enhancements can depend on the used date varieties. Al-Hooti and his colleagues  used fouOther formulations of date bars using skim milk powder at different concentrations and using other ingredients were also explored. The addition of 20% skim milk powder, soybean flour, or almond flour was explored . Between−1); in the early stages of date maturity, vitamin C can be found in high amounts, but it shows a decline during maturation [On the other hand, the date fruit used in  had a goturation . The addturation . The addturation ; chickpeturation  used roaaW). An aW below 0.6 is critical in manufacturing food products to ensure limited microbial growth. Munir et al. [w values ranging from 0.56 to 0.59 depending on the oatmeal concentration. The lowest aW was recorded with a higher amount of oatmeal. Lower moisture content can also affect the bars’ texture; an increase in the bars’ hardness to an acceptable range was observed. In contrast, the use of chickpea flour in other studies led to higher moisture content, resulting in extremely soft date bars compared to date bars containing corn flour. The latter was more liked in terms of texture and overall sensory acceptability [Adding cereals such as oatmeal can also significantly decrease the moisture content, leading to higher shelf-life stability by limiting microbial growth. Such ingredients can decrease the moisture content by binding water to their fractions, leading to lower water activity  were also explored. However, the latter protein sources could significantly affect the bars’ sensory attributes. They must be applied under the studied concentrations, adding other taste-intense ingredients such as nuts or dried fruits. Furthermore, adding dry ingredients such as cereals or legumes can enhance protein quantity and availability. For instance, oat, wheat, barley, soybean, or chickpea flours can enhance the amino acid profile. Cereals can also contribute to significantly higher fiber content due to their richness in good-quality fiber such as β-glucan. In addition, relatively high amounts of minerals and vitamins can be achieved with cereal addition.Cereal incorporation can not only promote significant enhancements in the nutritional attributes of the date bars but also result in significantly more enhanced texture and a more stable shelf life due to its ability to absorb moisture, leading to microbial growth limitation. On the other hand, incorporating other ingredients into date bars can not only target protein fortification or nutritional value in general; significant enhancements in the presence of bioactive or functional components can also be achieved. Functional ingredients such as extracts of fruits, plants, or seeds were shown to significantly increase phytochemicals such as phenolic acids or flavonoids, leading to a major increase in the antioxidant potential of the date bars. Although date fruit is considered rich in such bioactive components, which was one of the main reasons for its usage in developing energy bars, incorporating other nutritional ingredients can effectively boost dates’ antioxidant potential, providing an exceptionally functional product.From another point of view, sensory attributes are key factors in the acceptance of any food product. Adding taste-intense ingredients such as nuts or dried fruits, besides their positive effects on nutritional value, can improve the date bars’ sensory acceptability regarding taste, flavor, odor, and color. Seasoning using different spices such as cardamom, cinnamon, or ginger and adding salt can strongly impact the taste and flavor. Apart from choosing between the different suitable and functional ingredients regarding sensory enhancements, processing techniques can also promote significant enhancements in the bars’ sensory attributes. Roasted ingredients such as cereals, dried fruits (mainly coconut flakes), and nuts significantly boost the sensory attributes regarding odor, taste, and flavor. However, applying high temperatures may affect the bioavailability of some micronutrients; therefore, such techniques should be performed with suitable periods and temperatures.The high nutritional value of the prepared functional date bars presented in the previous section may play a strong part in promoting general health; in a recent pilot study, chronic kidney disease patients were given functional bars containing dates and other nourishing ingredients daily in combination with adherence to regular physical activity three times a week for three months; their measurements showed significant improvements in blood pressure levels, lipid metabolism, and other functional tests as compared to the control group, although those who consumed a functional bar daily with no weekly regular physical activity did not show such positive results, indicating the key impact of physical activity; however, the latter group had lower total cholesterol levels at baseline compared to other groups, indicating a possible explanation for the null results . NonetheThis review summarizes studies showing the potential application of date fruit for developing highly nutritious snack bars. The main target of developing date-based bars was protein enrichment, as dates are high in carbohydrates and low in protein. Skim milk powder represented the most common and favorable protein source, with positive effects and no negative alterations in sensory attributes. Other protein sources, such as cheese and plant-based proteins, were also found to enhance the protein content but may negatively affect the sensory attributes. Their addition must be used under the studied concentrations while incorporating taste-intense ingredients such as nuts  or dried fruits .Furthermore, protein enrichment is not the only target of developing date-based bars; providing an excellent functional snack bar can also be achieved. The reported studies have also used different nutritional and functional ingredients and showed significant enhancements in the presence of bioactive components, such as phenolics and flavonoids. However, the usage of the date fruit itself is promising in the snack bar industry due to its richness in various bioactive components. Dates’ bioactive components were demonstrated to promote several health-beneficial effects. Incorporating other nutritive ingredients besides dates can effectively boost dates’ functional properties. Among the different nutritious ingredients used in the reported studies in this review were extracts of fruit peels, plant seeds, or plant leaves. Such ingredients promoted significant enhancements in the bars’ functional and antioxidant potential. Moreover, the usage of date fruit wastes, such as date pits—seeds—was also shown to promote significant enhancements regarding nutritional and functional attributes.This review can provide a background for future approaches to developing balanced, nutritious, and highly functional date-based bars with highly acceptable sensory attributes for consumers, including older adults, children, and athletes. Considering that the global snack bar market has shown remarkable growth trends, such products would be highly accepted, and thus, scaling up is strongly recommended.


This study aimed to investigate the active composition and mechanism of the Shuganfang (SGF) in treating drug-induced liver injury (DILI) using network pharmacology and molecular docking. The potential active ingredients and targets of SGF were obtained from the Traditional Chinese Medicine Systems Pharmacology Database (TCMSP) database. DILI-related targets were queried from various databases including GEO, GeneCards, OMIM, NCBI, and DisGeNET. The STRING database was used to establish a protein-protein interaction (PPI) network. DAVID was utilized for conducting gene ontology (GO) function enrichment and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analyses. The data visualization and analysis of herb-ingredient-target and disease-pathway-target-ingredient networks were conducted using Cytoscape software (version 3.7.2). PyMoL and AutoDock software was used to select the best binding target for molecular docking. A total of 177 active ingredients,126 targets and 10112 disease targets were obtained, including 122 intersection targets. The identified potential active ingredients consisted of quercetin, kaempferol, luteolin, tanshinone IIa, nobiletin, isorhamnetin, beta-sitosterol and naringenin. The core targets implicated in the study were IL6, estrogen receptor 1 (ESR1), hypoxia-inducible factor alpha subunit 1 (HIF1A), MYC and vascular endothelial growth factor A (VEGFA). KEGG analysis revealed that the treatment of DILI with SGF mainly acted through apoptosis, the PI3K-Akt signaling pathway, and the tumor necrosis factor (TNF) signaling pathway. Furthermore, the binding affinities between the potential ingredients and the core targets were subsequently confirmed through molecular docking experiments. The findings indicated that the docking outcomes remained consistent and demonstrated a favorable capacity for binding. SGF exerts a therapeutic effect on DILI through multiple active ingredients, multiple targets and multiple pathways. Our findings contribute to a positive investigation and establish a theoretical basis for further extensive exploration of SGF as a potential treatment for DILI in future research. DILI is associated with severe adverse outcomes such as hepatitis, liver fibrosis, liver failure, and mortality. DILI can present with a hepatocellular, cholestatic or mixed pattern of disease.,5 These various forms of liver injury may necessitate distinct clinical approaches and treatment strategies. Despite the sensitivity of currently used liver parameters, the exact pathophysiology and specific therapies for DILI remain unclear. Therefore, there is an urgent need to develop novel medications for DILI that possess both safety and efficacy.Drug-induced liver injury (DILI) pertains to the aberrant results of liver function tests attributed to the medicinal compounds, pharmaceuticals, dietary supplements, and botanical substances. The estimated occurrence of DILI is between 14 and 19 instances per 100,000 people. Notably, the utilization of TCM in the treatment of DILI has yielded unique benefits and significant breakthroughs exemplified by the successful application of Hugan tablets,Schisandrae Chinensis Fructus. Shuganfang (SGF), a classical TCM formula, show significant effects on treating DILI. It is mainly comprised of Radix Bupleuri , Sedi Herba , Paeoniae Radix Alba , Schisandrae Chinensis Fructus , Atractylodes Macrocephala Koidz , Aurantii Fructus , Radix Salviae , licorice . Clinical investigations have demonstrated that SGF has significant therapeutic effect and few adverse in the treatment of DILI. However, there is still a need to elucidate the intricate mechanism of SGF in liver protection.In recent years, traditional Chinese medicine (TCM) has growing interest due to its safety and multifaceted impacts. Consequently, TCM has gained significant recognition as a viable approach for preventing and treating contemporary diseases. Network pharmacology offers a novel approach to uncover the underlying mechanisms of complex systems, encompassing multiple components such as TCM. In this approach, database screening, computer simulation, and information mining techniques are employed to acquire essential targets, pivotal pathways, and mechanisms of action, thereby facilitating a comprehensive elucidation of the intricate biological mechanisms underlying complex diseases and the molecular-level effects of pharmaceutical interventions. TCM exhibits its therapeutic effects by modulating various biological processes through the interaction of multiple targets and bioactive components. In recent years, network pharmacology has emerged as a valuable tool for investigating the intricate mechanisms underlying the efficacy of TCM in complex systems.–16Network pharmacology is a multidisciplinary interdisciplinary that has arisen under the umbrella of big data in biomedicine and artificial intelligence.In this particular study, network pharmacology and molecular docking analysis were utilized to predict the bioactive ingredients, potential molecular targets, and signaling pathways associated with the hepatoprotective effects of SGF against DILI. The results obtained from this research significantly contribute to a more comprehensive comprehension of the pharmacodynamics and mechanisms of action of SGF in the management of DILI. The study workflow is illustrated in Figure https://old.tcmsp-e.com/tcmsp.php). The TCMSP platform provided crucial information on the constituents of commonly used Chinese herbal medicines, as well as relevant parameters related to their absorption, distribution, metabolism, and excretion (ADME). Subsequently, the active ingredients that satisfied the criteria of oral bioavailability ≥ 30% and drug-likeness ≥ 0.18 were selected for subsequent target prediction.,18 In addition, the corresponding targets of these active ingredients in SGF were obtained and compiled from the TCMSP database. The obtained targets name was converted to the official gene symbol using the UniProt protein database (https://www.uniprot.org). The active ingredients and corresponding targets were uploaded to the Cytoscape 3.7.2 to construct the herb-ingredient-target network.The active ingredient information of SGF  were obtained from Traditional Chinese Medicine Systems Pharmacology Database , Series: GSE54255 . The “limma” package was used for identifying DEGs with logFC > 1 and P < .05. The R packages ggplot2 and heatmap were utilized to visualize the DEGs results. In addition, the key words“drug-induced liver injury” were searched in GeneCards (http://www.genecards.org), OMIM database (http://www.omim.org/), NCBI database (https://www.ncbi.nlm.nih.gov/gds/), and DisGeNET database (http://www.disgenet.org/) for the targets related to DILI. Finally, the DILI-related targets library was established by eliminating duplicate targets. Targets from SGF and DILI-related targets were intersected to determine intersections using Venny 2.1.0.The differentially expressed genes (DEGs) related to DILI were identified from Gene Expression Omnibus . The analysis results were filtered based on the following criteria: P < .05. Molecular function (MF), cellular component (CC), and biological process (BP) are all included in the GO enrichment analysis. R software was used to visualize the top 10 GO and top 20 KEGG pathway results with the lowest P value. The top 20 pathways and their corresponding targets and ingredients were sorted out and the disease-pathway-target-ingredient network was constructed using Cytoscape3.7.2.The GO and KEGG pathway enrichment analyses were carried out using the Database for Annotation, Visualization and Integrated Discovery . In this specific process, all proteins were dehydrated, the original ligands were extracted, and stored separately. Subsequently, the ligands and proteins were imported into AutoDockTools 1.5.6 in PDBQT format, and a docking grid box was constructed. Finally, the docking and identification of the optimal construct were performed using AutoDock Vina 4.2. PyMOL and Discovery Studio 2020 software were utilized for visualization of ingredients and proteins molecular docking.Further molecular docking verification was conducted on the top 8 ingredients identified from the herb-ingredient-target and the 6 hub targets obtained from the PPI network. AutoDock Vina 4.2 and AutoDockTools 1.5.6 were employed for conducting molecular docking analysis.There are 12 ingredients for Chaihu, 5 ingredients for Chuipencao, 7 ingredients for Wuweizi, 7 ingredients for Baishao, 7 ingredients for Baizhu, 60 ingredients for Dansheni, 5 ingredients for Zhiqiao, and 88 ingredients for Gancao. A comprehensive collection of 177 active ingredients of SGF was obtained subsequent to the elimination of 14 duplicated ingredients. Through an extensive search of the TCMSP database, we successfully identified 126 targets that corresponded to the aforementioned 177 SGF ingredients. The herb-ingredient-target network, which encompassed the 177 active ingredients and the 126 targets, consisted of 302 nodes and 2201 edges . Figure ,35 The identification of interventions capable of treating DILI and impeding its progression into liver failure would greatly benefit public health. At present, TCM has been scientifically proven to exhibit notable therapeutic efficacy for liver diseases through its multi-component and multi-target pharmacological effects. Shuganfang (SGF), a TCM formulation, has been extensively practiced in clinical settings for the treatment of liver diseases over a considerable period of time. Nevertheless, a thorough understanding of the active constituents and molecular mechanisms underlying the effects of SGF remains elusive.Drug-induced liver injury is a prevalent clinical issue that poses a substantial risk of morbidity and mortality. Nevertheless, there is currently a lack of effective treatments for DILI.,38 This approach enhances the ability to predict and categorize diverse therapeutic targets and interactions. Molecular docking-based virtual drug screening has become an integral aspect of structural biology. This study effectively employed network pharmacology and molecular docking methodologies to conduct a comparative analysis of mechanisms, with the objective of elucidating and identifying the constituents and hepatoprotective mechanism of SGF.Network pharmacology, an interdisciplinary field integrating molecular biology, genetics, and computer science, is employed to gather, analyze, extract, and apply biological information. Kaempferol has been found to possess the ability to inhibit hepatocyte apoptosis, thereby providing protection against liver failure in mice. This protective effect is achieved through the regulation of the ER stress-Grp78-CHOP signaling pathway. It has been demonstrated that luteolin exhibits antioxidative, anti-inflammatory, and anti-endoplasmic reticulum stress properties in mitigating acetaminophen-induced liver injury in mice. Furthermore, prior administration of tanshinone IIA has has been demonstrated to effectively safeguard the liver against APAP-induced hepatic injury through the activation of the Nrf2 pathway. Isorhamnetin has been found to effectively mitigate liver injury induced by acetaminophen through the reduction of oxidative stress, inflammation, and pyroptosis.The active ingredients of SGF were identified through a comprehensive search of the TCMSP database. Subsequently, a herb-ingredient-target network was constructed and analyzed, resulting in the identification of the top 10 active ingredients, which include kaempferol, luteolin, quercetin, tanshinone IIa, nobiletin, isorhamnetin, beta-sitosterol, naringenin, 7-Methoxy-2-methyl isoflavone, and isocryptotanshi. Research has demonstrated that quercetin possesses the capacity to enhance the expression of Nrf2/HO-1, thereby ameliorating acute liver injury in rats. Recent studies indicates that IL-6 can provide protection against acute liver injury (AILI) by directly signaling through IL-6R on hepatocytes, The IL-6/IL-6R signaling pathway activates the STAT3 pathway, leading to the inactivation of caspases and a reduction in reactive oxygen species. ESR1 has been extensively characterized in the human liver. Abnormal expressions of ESR1 in the liver have been implicated in the stimulation of hepatocyte injury and may serve as inducers or promoters of liver disease. HIF-1α, the master regulator of oxygen homeostasis, plays a crucial role in maintaining oxygen balance in response to hypoxia, inflammation, and oxidative stress. In APAP-induced liver injury, HIF-1α exhibits dual functions, promoting early damage and providing protection later in the pathogenesis. VEGFA is an endothelial cell mitogen and an important stimulator of sinusoidal endothelial cell proliferation. VEGF has a potent anti-apoptotic effect on hepatocytes through cell-cell interaction between sinusoidal endothelial cells and hepatocytes.The PPI network was subjected to topology analysis, resulting in the identification of key targets such as IL6, ESR1, HIF1A, MYC, and VEGFA. IL-6, IL-6, a multifunctional cytokine, is implicated in inflammatory responses and cellular immunity, and it plays a pivotal role in the restoration of hepatic function following liver injury. Apoptosis plays a critical role in the development of various liver diseases, such as cholestatic liver injury, alcoholic hepatopathy, diabetic hepatopathy, and drug-induced liver injury. The P53-upregulated modulator of apoptosis (PUMA) holds significant importance in mitigating liver injury induced by APAP. Additionally, Zhishi demonstrates hepatoprotective effects against APAP-induced liver necrosis by inhibiting PUMA.,56 The TNF signaling pathway plays a crucial role in various physiological and pathological processes, including regulation of immune reactions, cell proliferation, apoptosis, and inflammation induction. Numerous drugs have the capability to enhance inflammation and cytokine release, rendering liver cells susceptible to the cytotoxic impacts of cytokines like TNF. pretreatment with American ginseng berry has been found to mitigate APAP-induced liver injury by suppressing oxidative stress and inflammation responses through the TNF- α -mediated signaling pathways. The PI3K/AKT pathway has been identified as a crucial player in liver regeneration post-injury. exerts its hepatoprotective effects by regulating the expression of downstream target proteins, either inhibiting or enhancing their expression.,60 Research has demonstrated that leonurine effectively alleviates DILI by modulating the PI3K/ AKT signaling pathway.The KEGG pathway enrichment analysis demonstrates a strong correlation between the mechanisms of SGF in mitigating DILI and various biological processes, including apoptosis, the TNF signaling pathway, and the PI3K-Akt signaling pathway. Hepatocellular apoptosis is present during the early phases of human liver failure.The molecular docking findings have provided additional evidence that the primary active constituents of SGF possess robust binding affinity towards crucial targets linked to drug-induced liver injury (DILI). Specifically, luteolin exhibits strong binding activity with ESR1, kaempferol with ESR1, and tanshinone iia with IL6, thus indicating their substantial binding capabilities. The findings from molecular docking analysis provide additional evidence supporting the potential of the chemical constituents found in SGF as promising therapeutic agents for DILI. Nevertheless, it is important to acknowledge the limitations inherent in this study. Considering the impact of diverse subjective and objective factors, it is crucial to undertake subsequent animal experiments and clinical trials to authenticate the predictive results.It is important to acknowledge the limitations of this study. Firstly, the active compounds of SGF were obtained from literature and databases without experimental verification using LC/MS technology. Secondly, additional pharmacological experiments are required to validate the therapeutic mechanism of SGF on drug-induced liver injury (DILI).By employing a combination of systematic pharmacology and molecular docking techniques, our research has provided preliminary insights into the target action pathway and molecular mechanism underlying the therapeutic effects of adding and modified SGF in the treatment of DILI. Our study findings have verified that SGF exhibits efficacy in treating DILI by engaging various active constituents, targeting multiple sites, and influencing diverse biological pathways. The primary active components identified in SGF include luteolin, kaempferol, and tanshinone iia, while the key targets implicated in its therapeutic action encompass IL6, ESR1, HIF1A, MYC, and VEGFA. Furthermore, it is plausible that SGF elicits its therapeutic effects via the initiation of apoptosis, adjustment of the TNF signaling pathway, and activation of the PI3K-Akt signaling pathway. In conclusion, our findings establish a theoretical framework for conducting in vitro and in vivo investigations, as well as clinical utilization of SGF in the management of DILI.Conceptualization: Ying Wang, Yang Ye.Data curation: Xueying Chen, Yan Wang, Hong Zhong.Funding acquisition: Ying Wang, Xueying Chen, Liqin Liu.Methodology: Ying Wang, Xueying Chen, Yan Wang.Project administration: Liqin Liu, Yang Ye.Writing – original draft: Ying Wang, Hong Zhong.Writing – review & editing: Xueying Chen, Yang Ye.


During the last 20 years, molecular alterations have gained increasing significance in the diagnosis and biological assessment of tumors. Gliomas represent the largest group of tumors of the central nervous system, and the main aim of this review is to present the current knowledge on molecular pathways and their alterations in gliomas. A wide range of new insights has been gained, including evidence for the involvement of the WNT pathway or the hippo pathway in the pathobiology of gliomas, indicating a broad involvement of different pathways formerly not considered to play a central role in gliomas. Even new aspects of angiogenic, apoptotic, and metabolic pathways are presented, as well as the rapidly growing field of epigenetic processes, including non-coding RNAs. The two major conclusions drawn from the present review are the distinct interconnectivity of the whole spectrum of molecular pathways and the prominent role of non-coding RNAs, especially circular RNAs, in the regulation of specific targets. All these new insights are discussed, even considering the topic of the resistance to therapy of gliomas, along with aspects that are still incompletely understood, like the role of hydroxymethylation, or even ferroptosis, in the pathobiology of gliomas. Gliomas are the most frequent brain tumors, representing approximately 80% of all tumors of the central nervous system ,2. In re++ by disrupting its mobilization in vitro or in experimental models provided evidence for reduced tumor cell invasion and for the extended survival of animals with different tumor types, including glioblastomas [mice [mice [The overexpression of the tyrosine kinase receptors (RTKs) and their ligands is among the most frequent alterations in glioma tumorigenesis, leading to alternate autocrine or paracrine stimulatory loops and to the aberrant activation of RTKs and their downstream molecular pathways. The RTKs consist of a transmembrane domain, an extracellular ligand-binding domain, an intracellular tyrosine kinase domain, and tyrosine residues leading to downstream activation after their phosphorylation. Due to a variety of molecular changes, such as oncogenic mutations, the silencing of tumor-suppressor genes, gene amplification, or epigenetic changes, overactivated RTKs may contribute to the oncogenic phenotype. The effects of this dysregulation include the overexpression of growth factors or RTKs themselves, or even their involvement in autosomal genetic or epigenetic changes, such as mutations, chromosomal rearrangements, or promotor methylation ,3,7. Ovelastomas ,11. The lastomas ,13,14. Tlastomas ,16. Focaas [mice ,18. An ias [mice . Integrias [mice . Other ias [mice . Based oce [mice . Of notece [mice . Althougce [mice ,24,25,26NF1 encodes neurofibromin, which is a physiological suppressor of RAS, and NF1 mutations are responsible for the absence of this natural suppression. BRAF gene mutations such as BRAFV600E lead to overactivation of the MAPK pathway, which has led to successful attempts to target this mutation with dabrafenib or trametinib in children and adult patients [INK4a/CDK4/RB1 pathway often shows alterations in gliomas. Inactivation of pRB1 through its phosphorylation mediated by CDK4/6 is one way to promote tumor progression. This activation of the cell cycle transition from G1 to the S phase is supported by an increased release of the transcription factor E2F. p16INK4a and even cyclin-dependent kinase 2 types A and B (CDKN2A/B) are regulators of CDK4/6-mediated pRB1 phosphorylation so that the loss of normal function of pRB1 can be triggered by alteration of either of these proteins. The interconnectivity of all these pathways is confirmed by the fact that key molecules such as CDKN2A/B are involved in the p16INK4a/CDK4/RB1 pathway and the TP53/MDM2/MDM4 pathway [Drosophila, and it remains to be determined whether this also applies to other species [Overactivation of receptors by their corresponding ligands or also by alternative or pathological stimuli such as mutations leads to the increased stimulation of various signaling pathways that contribute to the pathogenesis and progression of gliomas. All of these signaling pathways have a complex network of connections, confirming their role in numerous cellular processes. A good example is the PI3K/AKT/PTEN pathway, which is involved in several processes such as proliferation, glucose metabolism, or cell survival. The first step within the activated pathway is the recruitment of phosphatidylinositol 3-kinase (PI3K), which phosphorylates phosphatidyl inositol bisphosphate to triphosphate PIP2 to PIP3). Normally, PTEN (phosphatase and tensin homologue) blocks PiP3 signaling and AKT (protein kinase B) activation, inhibiting proliferation. Frequent PTEN mutations in glioblastomas lead to a blockade of this inhibition and a complete activation of this signaling pathway with its main effector molecule, the serine/threonine kinase mTOR . mTOR itself leads to a promotion of cell growth and cytoskeletal organization through the activation of the multiprotein complexes, mTORC1 and mTORC2 ,28,29. T to PIP3.patients . The tumpatients ,32,33. T pathway ,34. The  pathway . Its cen pathway ,37. Diff pathway . VPS25 i pathway . One inh pathway . The inh species . The NOT species . In glio species . This si species ,42,43. T species ,45,46,47 species . The hip species ,50. Invo species . The deg species ,50. The  species ,52. As t species ,50.gene for telomerase reverse transcriptase (TERT) that lead to TERT overactivation, thereby preventing tumor cells from apoptosis. TERT mutations are found in many glioblastomas, whereas mutations of the X-linked gene of α-thalassemia (ATRX), leading to inactivation of the ATRX protein and induction of ALT, are found in many IDH-mutated astrocytomas. Of note, TERT mutations and ATRX mutations are mutually exclusive in gliomas, and the exact mechanism of how ATRX mutations, in particular, induce ALT in gliomas is still under investigation [UV irradiation resistance-associated tumor-suppressor gene (UVRAG) [The angiogenic pathways in tumors are highly interconnected, as the activation of factors also leads to the activation of other downstream signaling pathways that promote tumorigenesis. The most important pro-angiogenic factor is vascular endothelial growth factor receptor (VEGFR), but other factors also contribute to tumor angiogenesis, such as fibroblast growth factor receptor (FGFR), angiopoietins 1 and 2 (Ang1 and Ang2), erythropoietin (EP), transforming growth factor beta (TGF-β), and matrix metalloproteinases (MMPs) . The inctigation ,73,74,75tigation  and eventigation . Another (UVRAG) . A major (UVRAG) . mice, confirming the importance of glucose and glutamine pathways in the growth of malignant gliomas and providing a potential therapeutic strategy [EGFR gene with deletion of exons 2-7, present in the majority of glioblastomas with EGFR amplification. An important effect of EGFRvIII is the activation of heterogeneous nuclear ribonucleoprotein A1 (hnRNP-A1), which induces the formation of the protein DeltaMax by splicing the Myc-interacting protein Max. DeltaMax is a potent promoter of glycolysis and tumor growth in vivo, and a study of 131 glioblastoma patients showed a significant correlation between increased expression of hnRNP-A1 and glycolytic genes with shorter overall survival [An important alteration in metabolism, especially in highly proliferating gliomas, is that tumor cells meet their energy requirements by glycolysis instead of oxidative phosphorylation. This phenomenon is known as the Warburg effect and results in a high rate of glycolysis in proliferating tumor cells, including an overproduction of lactate. In the absence of significant efflux, increased cellular lactate inhibits glioma invasion. Therefore, accelerated lactate efflux is necessary to promote tumor cell invasiveness . This lastrategy . In IDH-strategy . Under pstrategy ,85. Disrstrategy . Anotherstrategy ,88. One strategy . Tumor dsurvival . In addisurvival ,92. Moresurvival . The impsurvival . Homo sapiens are certainly still largely unknown. So far, the focus has been on investigating the DNA methylation profile in diseases and even gliomas. In gliomas, DNA methylome analysis can be performed using bisulfide PCR or even novel strategies, such as nanopore sequencing, but most clinical applications are still based on chip arrays with a readout of more than 800,000 methylation sites. A statistical comparison of the results with thousands of reference samples from the “Classifier of Molecular Neuropathology” [forkhead box A2 gene (FOXA2), which plays a role in prostate cancer and whose promoter methylation leads to activation of the FOXA2 gene expression [The term epigenetics refers to molecular processes involved in the regulation or alteration of gene expression that are not associated with a change in gene sequence. The three most important epigenetic processes are methylation, histone alterations, and regulation by so-called “non-coding RNA molecules”. The molecular basis for the methylation of DNA or other biomolecules are methyltransferases classified according to the ‘Enzyme Nomenclature Database’ of subclass 2.1.1 . DNA metthology”  contributhology” . Althougthology” ,99. The thology” ,100. Sevpression ,102. Tempression . For othpression . The rolpression . Overexppression ,106,107.pression ,110,111.pression . This hipression , but it pression ,115. Anopression ,116. Nonpression ,118. Thepression ,119,120.pression . Since tpression ,50,121. pression ,123,124,pression ,126  is widely accepted ,146,147.The distinct interconnectivity of molecular cellular signaling pathways in terms of their involvement in tumorigenesis and glioma growth due to dysregulation and alterations is an important recent finding in glioma research. The second most important discovery in recent years is the recognition of the significant impact of the group of non-coding RNAs (ncRNAs) on molecular pathobiology in tumors, as it appears that ncRNAs, especially circular RNAs (circRNAs), have an overriding influence on the regulation of molecular signaling pathways in gliomas. Most circRNAs are formed by circularization of exon sequences, and only a few circRNAs are formed from introns . Most ciRegarding tumor angiogenesis in gliomas, the search for targeted fixed points is currently an important goal, and in particular, experimental progress has been made by simultaneously targeting VEGF and Ang2 in a mouse model with prolonged animal survival . There aA particular advance in recent years has been the introduction of molecular criteria for the diagnosis and/or classification of specific tumor entities since the fourth edition of the WHO classification of central nervous system tumors in 2016 . Since t


In this study, the potential active ingredients and mechanisms of action of LGZG decoction in the treatment of type 2 diabetes were initially investigated, which provided a basis for the in-depth study of its drug basis and mechanisms of action.To investigate the mechanism of action of the classical formula Ling-Gui-Zhu-Gan (LGZG) decoction in treating type 2 diabetes mellitus based on network pharmacology and molecular docking. The active ingredients and targets of LGZG decoction were collected by the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform database and mapped using Cytoscape software to show their interrelationships. GeneCards, Pharmacogenomics Knowledge Base, OMIM, Therapeutic Target Database, and Drugbank databases were used to obtain targets related to type 2 diabetes; protein-protein interaction networks were established with the help of the STRING platform. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses were performed on selected core targets with the help of the Metascape platform. Finally, the AutoDock platform was used to perform molecular docking and display the results by Pymol software. One hundred twenty-one active ingredients, 216 effective target genes, 11,277 type 2 diabetes mellitus-related genes, 210 crossover genes, and 18 core genes were obtained for LGZG decoction. The results obtained by Kyoto Encyclopedia of Genes and Genomes indicated that the advanced glycosylation end products-receptor of advanced glycosylation end products signaling pathway, the phosphatidylinositol 3 kinase-Akt signaling pathway, and HIF-1 signaling pathway might be the key signaling pathways. Molecular docking showed that the binding energy of quercetin, kaempferol, naringenin, and licorice chalcone A to the core target genes were all <5.0 kJ-mol According to the International Diabetes Federation, the number of people with diabetes is expected to increase to about 700 million by 2045. It has been documented that type 2 diabetes mellitus (T2DM) accounts for about 95% of diabetic patients, making it a global health problem. T2DM is a non-insulin-dependent type of diabetes, which can induce various complications, including diabetic nephropathy and cardiovascular. The clinical treatment of T2DM mainly includes lifestyle and pharmacological interventions. However, the long-term application of simple hypoglycemic drugs is often accompanied by different toxic side effects, such as heart failure and increased risk of fracture in women.,5Diabetes mellitus is a group of chronic hyperglycemic metabolic diseases caused by defective insulin secretion or utilization, with polydipsia, polyphagia, polyuria, and weight loss as typical clinical manifestations.,7 which has the advantages of less adverse reactions and stable efficacy and is receiving more and more attention from medical practitioners. The etiology of diabetes mellitus belongs to the category of “emaciation-thirst disease” in Chinese medicine. It is generally believed that the etiology of thirst is based on yin deficiency, dryness, and heat. The disease is usually located in the lung, stomach, and kidneys. However, the clinical etiology is complex and varies, and it is not uncommon to have atypical symptoms and spleen deficiency with dampness.–12In contrast, the treatment of T2DM in traditional Chinese medicine (TCM) can make up for the shortcomings of Western medicine through its holistic concept and evidence-based treatment. It can regulate blood glucose by improving pancreatic β-cell function, promoting insulin secretion, and improving insulin sensitivity,In this paper, we use network pharmacology and molecular docking methods to predict the core target genes of Ling-Gui-Zhu-Gan (LGZG) decoction for the treatment of T2DM and investigate the potential pathway mechanisms to pave the way for further research. The detailed workflow of this study is presented in Figure http://tcmspw.com/tcmsp.php). The chemical compositions of “Fu Ling,” “Gui Zhi,” “Bai Zhu,” and “Gan Cao” were searched. Based on the absorption distribution and metabolic excretion screening conditions, with oral bioavailability ≥ 30% and drug similarity ≥ 0.18, the active ingredients and potential targets of Fu Ling, Gui Zhi, Bai Zhu, and Gan Cao in LGZG decoction were obtained. The obtained compound target genes were entered into the Uniprot (https://www.uniprot.org/) database, normalized, and transformed to find the corresponding gene symbols (standard gene names).The chemical composition of “Fu Ling,” “Gui Zhi,” “Bai Zhu,” and “Gan Cao” was retrieved from the database of Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) (http://www.genecards.org/), OMIM (http://www.omim.org/), Pharmacogenomics Knowledge Base (https://www.pharmgkb.org), Therapeutic Target Database (http://db.idrblab.net/ttd), and DrugBank (https://www.drugbank.ca) databases for the keyword “type 2 diabetes mellitus.” The search results of the 5 databases were combined, all disease-related genes were merged, and duplicate target genes were removed to obtain the potential pathogenic targets of T2DM.The GeneCards , the study species was limited to “human,” the confidence level was set to 0.9, the free nodes were hidden, and the rest of the parameters were set to default to obtain the PPIs, and exported to Save as TSV format file and save the original image. The resulting files were imported into Cytoscape 3.8.0 software for visualization. The CytoNCA plug-in was used to calculate the Betweenness, Closeness, Degree, Eigenvector, Network, and local average connectivity-based scores for each node. Each node is scored, and the node with a higher score is retained by filtering the value > median value to obtain the core gene of the network.Protein-protein interaction (PPI) studies the correlation between compounds and disease-related protein molecules from the perspective of biochemical, signal transduction, and genetic networks. The common target genes of LGZG decoction and T2DM were imported into the STRING database , and its water molecules and small molecule ligands were removed by PyMOL software. The resulting small molecule ligand and protein receptor files were converted into PDBQT format, and the active pockets for molecular docking were determined simultaneously. Finally, molecular docking was performed using Vina software.The above analysis selected top-ranked active ingredients and core genes, and molecular docking was verified according to whether there was an interaction between the core compounds and the core target genes. 2D structure files of small molecule ligands were downloaded from the PubChem database, and the 2D structure was converted to a 3D structure in ChemOffice software and saved in mol2 format. The pdb format of protein 3D structure was downloaded from the Protein Data Bank database .As shown in Table As shown in Figure http://links.lww.com/MD/I645, which illustrates the basic information of network construction and the types of nodes).Figure The above 210 intersecting genes were uploaded to the STRING database to obtain the PPI map, as shown in Figure P value was set at 0.05. The GO annotation analysis was performed in 3 aspects: biological process (BP), cellular component, and molecular function (MF). The results of GO annotation analysis showed that the BP mainly involved the response to xenobiotic stimuli, the response to metal ions, the cellular response to chemical stress, the response to lipopolysaccharide, peptide, and extracellular matrix, and the oxidative stress response. Composition mainly involves membrane rafts, membrane microstructure domains, transcriptional regulatory factors, transcriptional regulatory complexes, protein kinase complexes, vesicle lumen, etc. MFs mainly involve DNA-binding transcription factors, cytosolic receptor activity, activated ligands, ligase conjugates, subtilisin conjugates, G protein-coupled amine receptor activity, cytosolic steroid receptor activity, etc.The GO analysis was performed in R language for the intersection of LGZG decoction and T2DM Fig. , and theP to .05 .The R software was used to analyze the pathways and visualize them by setting As shown in Table  Its derivatives have a protective effect on diabetic neuropathy by inhibiting the Wnt/β-catenin signaling pathway. Quercetin intake was negatively correlated with the prevalence of T2DM, and its normalization of pancreatic β-cells was achieved by reducing iron death in T2DM by eliminating oxidative stress. Kaempferol modulates M1/M2 phenotype under hyperglycemia and indirectly protects against podocyte apoptosis by regulating macrophage M1/M2 differentiation; in terms of its hypoglycemic mechanism, it may achieve hypoglycemic efficacy by inhibiting the digestion of carbohydrates. Another scholar found in rat experiments that naringin could prevent hyperglycemia-mediated-induced inflammation and damage to liver and pancreatic tissues by blocking the activation-mediated anti-inflammatory effects of NF-κB and its down-regulated genes, including pro-inflammatory cytokines. In contrast, licochalcone A is capable of modulating insulin sensitivity in modern pharmacology, thus exerting a corresponding hypoglycemic effect.Our findings indicate that the top-ranked active ingredients in LGZG decoction were quercetin (quercetin), kaempferol (kaempferol), naringenin (naringenin), and licochalcone a . It has been shown that quercetin inhibits NOX expression and ROS production in rat Achilles tendons under hyperglycemic conditions, has antioxidant and anti-inflammatory effects, and can prevent the development of diabetic tendinopathy.The GO functional enrichment analysis results showed that the treatment of T2DM by LGZG decoction is extremely complex, involving multiple aspects of BPs, cell composition, and MFs. To sum up, the action targets located at the center of the network constructed by Cytoscape include STAT3, HIF1A, STAT1, AKT1, and CCND1.–23 STAT3, under high glucose conditions, after phosphorylation, causes hepatic gluconeogenesis while reducing glycogen synthesis and elevating blood glucose. In experiments in which systemic injection of streptozotocin-induced hyperglycemia in rats, it was found that during streptozotocin-induced diabetic retinopathy, the levels of HIF-1α, as well as the pro-inflammatory cytokines IL-1β, IL-6, and TNF-α, were increased and that HIF-1α led to upregulation of IL-6 and TNF-α and their receptors as well as Caspase-3, and inhibition of HIF-1α decreases the expression of the pro-inflammatory mediators IL-6 and TNF-αin diabetic retinopathy, thereby reducing the incidence of diabetic retinopathy. AKT1 is an important link in the PI3K/AKT/mTOR signaling pathway. Large amounts of AKT1 activate mTOR and enhance SREBP1 efficacy, thereby increasing intracellular triacylglycerol in tissues to achieve energy homeostasis. Akt is essential for insulin and nutrient-mediated regulation of hepatic metabolism in the body. It has also been shown that hepatic CCND1 deficiency leads to increased gluconeogenesis and, consequently, to hyperglycemia.It has been shown that in studying the effects of T2D risk genes and SNPs on transcriptional binding affinity, the top 4 transcription factors associated with enrichment were found to be Rfx1, Nkx2-5, NR2C2, and MZF15-13, while TF-Rfx1 is in the signal transducer and activator of transcription 3 (STAT3) pathway The expression of Rfx1 is regulated by the IL-6-STAT3 signaling pathway, which also indicates that STAT3 plays an important role in the pathogenesis of diabetes. and reduce insulin sensitivity to adipocyte cell membrane surface receptors, which will cause hyperlipidemia and atherosclerosis manifestations in the long-term and aggravate the risk of T2DM complications. One study showed that the risk of gestational diabetes mellitus increased 18.48-fold for each unit increase in atherosclerotic plasma index, indicating that reasonable lipid control in mid-pregnancy may reduce the incidence of gestational diabetes mellitus, statistically demonstrating an association between lipids and atherosclerosis and diabetes. The PI3K/Akt signaling pathway of key downstream factors is closely related to the regulation of glucose and lipid metabolism, and data suggesting a reduction in the occurrence of diabetic osteoporosis with further upregulation of PI3K/Akt-related protein levels. In turn, activation of the HIF-1 signaling pathway is associated with inflammatory and fibrotic processes in the renal unit and vascular calcification in patients with T2DM.,36The AGE-RAGE signaling pathway, PI3K-Akt signaling pathway, and HIF-1 signaling pathway in lipids and atherosclerosis, diabetic complications may play an important role in the treatment of T2DM with LGZG decoction as seen in the results of enrichment pathways in KEGG. Studies have shown that a persistent hyperglycemic state will cause plasma protein glycosylation, and insulin glycosylation can distort insulin signalingIn conclusion, the pharmacological process of LGZG decoction for the treatment of T2DM may be a synergistic combination of multiple active ingredients, multiple action targets, and multiple signaling pathways. Various core ingredients in the formula, such as quercetin, kaempferol, naringenin, and licorice chalcone A, may act on STAT3, HIF1A, STAT1, AKT1, CCND1, and other potential key targets and then exert synergistic effects on multiple signaling pathways such as AGE-RAGE signaling pathway, PI3K-Akt signaling pathway and HIF-1 signaling pathway in lipid and atherosclerosis, diabetic complications. In turn, it exerts a holistic and complex regulatory effect on multiple signaling pathways, including the AGE-RAGE signaling pathway, PI3K-Akt signaling pathway, and HIF-1 signaling pathway in lipid and atherosclerosis diabetes complications. According to TCM, T2DM belongs to the category of “thirst” in TCM and is often due to spleen deficiency and water retention. The combination of the 4 herbs in LGZG decoction brings out the effect of “warming and harmonizing” to achieve the function of warming yang, transforming qi, promoting water, and dispelling dampness. This paper predicts and analyzes the possible pharmacological mechanism of LGZG decoction in the treatment of T2DM by using software technology related to network pharmacology. At the same time, the results of the current experimental research are used to prove that the results are still scientific and reasonable. However, from the perspective of scientific rigor, further experimental research data are needed to support the results, so it provides a theoretical basis and direction to explore the pharmacological process of LGZG decoction in the treatment of T2DM. However, from the point of view of scientific rigor, in the study, although we have selected as much data as possible from the database, some important targets may still be missed because the database needs to be updated in time or the research needs to be more comprehensive. Moreover, the study that some of the selected signaling pathways are indeed part of the pathogenesis of diabetes and its complications is not enough. There are differences between in vitro theoretical study and in vivo metabolism. In addition, the study aims to construct a possible active ingredient-target network through relevant software technology. Due to the limitations of the research technology, we need further experimental research data to prop up.Conceptualization: Feng Long, Zhe Zhang.Data curation: Feng Long, Zhe Zhang.Methodology: Feng Long, Zhe Zhang.Resources: Feng Long, Chunxiu Luo, Jinlian Guo, Lin An.Software: Feng Long, Zhe Zhang.Supervision: Xiao Lei.Writing – original draft: Feng Long.Writing – review & editing: Zhe Zhang, Xiao Lei.


To identify the active constituents, core targets, immunomodulatory functions and potential mechanisms of Dizhi pill (DZP) in the treatment of myopia. The active constituents and drug targets of DZP were searched in the TCMSP, Herb databases and correlational studies. The targets of myopia were searched in the TTD, Genecards, OMIM and Drugbank databases. Gene expression profile data of GSE136701 were downloaded from the GEO database and subjected to WGCNA and DEG analysis to screen for significant modules and targets of myopia. Intersectional targets of myopia and DZP and core targets of myopia were analyzed through the String database. The GO and KEGG enrichment analyses of the interested targets were conducted. Cibersort algorithm was used for immune infiltration analysis to investigate the immunomodulatory functions of DZP on myopia. Autodock was used to dock the important targets and active constituents. Eight targets  and 5 active constituents  were identified from DZP. In pathways identified by the GO and KEGG enrichment analyses, “ATP metabolic process” and “AGE-RAGE diabetes complication signaling” pathways were closely related to the mechanisms of DZP in the treatment of myopia. Molecular docking showed that both the intersectional targets and core targets of myopia could bind stably and spontaneously with the active constituents of DZP. This study suggested that the mechanisms of DZP in the treatment of myopia were related to active constituents: Quercetin, Beta-sitosterol, Diincarvilone A, Ferulic acid methyl ester and Naringenin, intersectional targets: STAT3, PIK3CA, PIK3R1, MAPK1, MAPK3, and HSP90AA1, core targets of myopia: MIP and LGSN, AGE-RAGE signaling pathway, positive regulation of ATP metabolic process pathway and immunomodulatory functions. In addition to affecting the quality of life, severe myopia can further develop into more severe diseases such as glaucoma and cataract, which may even lead to blindness.–6 Current treatments for myopia include pharmacotherapy, keratoplasty and laser corneal surgery. However, the specific mechanisms of myopia development and myopia gene host reactions are still unclear.,8 Existing drug therapies, atropine eye drops most commonly, have photosensitive, anticholinergic and other side effects, and surgical treatment and laser treatment can cause some invasive side effects such as loss of vision, high myopia and retinal damage. Chinese botanical drugs (CBD) were traditionally used for myopia in China. They are considered relatively safe and noninvasive, and are now widely used in combination with acupuncture to improve myopia in children. Studies showed that CBD could improve retinal lesions and restore visual acuity and visual field.Myopia is the most common eye disease, According to traditional Chinese medicine (TCM), myopia is mostly caused by deficiency of liver and kidney yin and deficiency of Qi and blood, as stated in the TCM classic work Yixue Rumen and approved by modern TCM research: “Those who can see near but not far, or see one into two, are deficient in liver and kidney.” Studies on TCM constitutional theory showed that myopia patients were mostly Yin deficiency, and part of them were Qi-stagnation.,17 Among the ingredients of DZP, Sheng Dihuang and Tiandong can nourish Yin, Zhiqiao can transform Qi, and Juhua can comfort the liver and brighten the eyes. Therefore, Yin will be replenished, Qi stagnation will be eliminated, and vision will become clear. One study has reported a case of myopia cured by DZP and identified its efficacy in relieving myopia and improving blurred vision. However, the pharmacological mechanisms of DZP in the treatment of myopia are still unclear, which to a certain extent limit the application of DZP in clinic.Dizhi pill (DZP) is a CBD formula that was first presented in the Renzhai Zhi Zhi Fang, which mentioned that “Dizhi pill is indicated for the inability to see far away.” It consists of Rehmannia glutinosa (Gaertn.) DC. [Orobanchaceae] (Sheng Dihuang), Asparagus cochinchinensis (Lour.) Merr. [Asparagaceae] (Tiandong), Citrus × aurantiifolia (Christm.) Swingle [Rutaceae] (Zhiqiao), and Chrysanthemum arcticum L. [Asteraceae] (Juhua), and is a common formula for the treatment of myopia in Chinese medicine.The aims of this study are to identify the active constituents, core targets and potential mechanisms of DZP for myopia (including pathways and immune regulation functions), and to assess the stability of the molecular docking of the active constituents of DZP with the intersectional targets and specific targets of myopia. By network analysis and bioinformatics methods, we identified the active constituents and relevant targets of DZP in the treatment of myopia and screened myopia-specific modules and genes. We also explored the potential mechanisms of its action using the Gene Ontology (GO) and Kyoto Encyclopaedia of Genes and Genomes (KEGG) enrichment analyses and differences in immune infiltration levels between groups to verify the immunomodulatory functions of DZP in the treatment of myopia. The steps used to identify the mechanisms of DZP for myopia in this study are presented in Figure The network analysis materials and methodological steps of this study were performed in accordance with the Guidelines on Network Pharmacology Evaluation Methods.https://old.tcmsp-e.com/tcmsp.php) database, and screened according to the standards of oral bioavailability ≥ 30% and drug likeness ≥ 0.18.,21 As the ingredient Sheng Dihuang was not included in the TCMSP, the information relevant to its active constituents was searched through the Herb (http://herb.ac.cn/) database, and the studies relevant to its active constituents were searched through China National Knowledge Infrastructure, Wanfang and PubMed databases using the search strategy presented in File S1, Supplemental Digital Content, http://links.lww.com/MD/J488.–24 For the active constituents whose oral bioavailability and drug likeness were not available, the SMILES numbers of the active constituents were obtained from the Pubchem (https://pubchem.ncbi.nlm.nih.gov) database, and then the active constituents were screened through Swiss Target ADME (www.swissadme.ch) database according to the Lipinski filter principle and BOILED-Egg principle. After screening of the active constituents, the targets of the active constituents were predicted by the Swiss Target Prediction (www.swisstargetprediction.ch) database, and then the protein names were converted into their standard upstream gene names by the Uniprot (https://www.uniprot.org) database and the duplicated results were removed. These proteins and targets were tagged with their upstream gene names in the following sections of this study.The active constituents of the ingredients of DZP were searched through the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform , Online Mendelian Inheritance in Man (https://www.omim.org), Drugbank (https://go.drugbank.com), and Therapeutic Targets Database (http://db.idrblab.net) databases using the search term “myopia” and the duplicate results were removed.Myopia-related targets were obtained through the Genecards  between myopia and normal samples were screened according to the statistical significance of the between-groups difference (P < .05), with log2FC < -1 as down-regulated genes and log2FC > 1 as up-regulated genes. The significant DEGs among groups were selected in order of correlations of their Gene Counts between samples. The DEGs were shown using a heat map and a volcano map created by RStudio and the website “bioinformatics” (www.bioinformatics.com.cn).We searched “myopia” in the GEO database to obtain gene expression profile data GSE136701 of lens epithelium in high myopia and normal eyes. After clustering analysis to remove outlier gene expression data from each sample, the adjacency between genes was calculated by network topology analysis using the standard of R2 = 0.7 to screen the best soft threshold. The adjacency was transformed into a topological overlap matrix and the dissimilarity was calculated. The clustering tree diagram was drawn by hierarchical clustering method and the minimum number of module genes was set as 300, division sensitivity as 1, and distance of merging similarity modules as 0.3 to construct the weighted gene co-expression network. In the phenotypic correlation analysis, modules with the strongest positive and negative correlations between modules and the grouping factor were screened and the genes in the modules were selected.Weighted gene co-expression network analysis (WGCNA) was performed on myopia gene expression profile data using the “WGCNA” package of RStudio to screen the characteristic modules and genes of myopia. The TPM values of normalized counts values in myopia gene expression profile data were calculated to predict the differences in the proportions of immune cells between myopia and normal samples, and the immunomodulatory functions of the constituent ingredients of DZP on each immune cell in the treatment of myopia were validated by reviewing the correlational studies.The intersectional genes in the results of differentially expressed gene (DEG) analysis and WGCNA were screened and the immune infiltration analysis was performed using the CIBERSORT algorithm.The overlap targets between DZP and myopia and the overlap targets between DEG analysis and WGCNA were screened using Microsoft Excel. The intersecting targets were visualized using the JVenn plot from the Bioinformatics website.https://cn.string-db.org) database and the species was set as “Homo sapiens.” The minimum required confidence score was set as “highest confidence (0.900)” for the analysis of the intersectional targets of DZP and myopia and was set as “medium confidence (0.400)” for the analysis of the intersectional targets of DEG analysis and WGCNA. PPI networks were drawn using Cytoscape_v3.9.1 software based on the analysis results of String database. The degree centrality (DC) of each node protein in the PPI networks was calculated using the function “CytoNCA.”The intersecting targets were imported into the String , cellular component (CC), biological process (BP) – were derived and the results of the enrichment analyses were visualized using bubble plots.To identify which pathways were the most related to the intersectional targets, and to conclude the potential pathways relevant to effect of DZP and myopia development, the enrichment analyses were performed using the “clusterProfiler” package of RStudio. The species was set as “Homo sapiens,” and enrichment analyses were performed based on the GO and KEGG databases according to the selection standard www.pdbus.org) databases, and structures of active constituents were obtained by the Pubchem database. Autodock 1.5.7 was used to perform dehydration, hydrogenation, small molecule ligand energy minimization, acquisition of active pockets and molecular docking of active constituents and targets. According to the official Autodock recommendations, 50 molecular docking operations were performed for each pair of active constituent and target, and the binding activity between each pair of molecule and target was considered good when the molecular docking binding energy was equal or superior to 5 kJ/mol. The results were visualized using PyMOL 2.2.0 software.The top 5 DC-ranked active constituents in the results of DZP ingredients – active constituents – targets – myopia network were selected for molecular docking with the top 6 DC-ranked intersectional targets of DZP and myopia, and with the top 2 DC-ranked targets of myopia screened by WGCNA and DEG analysis. Protein structures were obtained by the Uniprot and RCSB PDB  originated from both Juhua and Zhiqiao, quercetin (A2) originated from both Juhua and Tiandong, and beta-sitosterol (B1) originated from all 4 ingredients . There were 412 predicted targets and 1027 additional targets obtained from relevant studies. A total of 758 targets were obtained after de-duplication. The information of the active constituents of DZP sorted according to their action targets is presented in Table We identified a total of 43 active constituents of DZP, including 17 from Juhua, 13 from Sheng Dihuang obtained from the Herb database and 3 correlational studies,A total of 5393 myopia targets were obtained from the Drugbank, Therapeutic Targets Database Online Mendelian Inheritance in Man and Genecards databases after de-duplication.http://links.lww.com/MD/J490), including 446 down-regulated genes and 395 up-regulated genes. The volcano plot of DEG analysis is presented in Figure A total of 23,348 genes were obtained after gene ID conversion using GPL570 platform , 245 intersectional targets of DEG analysis and WGCNA, and 2 (AR and TYRO3) intersectional targets of DZP, myopia, DEG analysis and WGCNA. The 181 intersectional targets of DZP and myopia were considered the functional targets of DZP in treating myopia, and 245 intersectional targets of DEG analysis and WGCNA were considered as core targets of myopia which may related to the development and occurrence of myopia.As mentioned above, we identified 5393 targets related with myopia, 758 potential targets of DZP, 841 targets corresponding to their up-regulated and down-regulated genes in DEG analysis, and 1617 targets of WGCNA results. The interrelationships of these targets are shown in the JVenn plot , including 199 pathways in MF, 99 pathways in CC, 2443 pathways in BP, and 171 KEGG pathways , were obtained from the 181 intersectional targets of DZP and myopia. The GO enrichment pathways with the highest number of enriched targets were “positive regulation of kinase activity” and “transmembrane receptor tyrosine kinase activity” pathways , and the KEGG analysis identified 13 pathways . Sorted by the numbers of targets on the pathways, the GO results were mainly related to “lens structural composition,” “cation channel complex” and “platelet activation” pathways  in corneal tissue thereby slowing down corneal scar formation and promoting corneal tissue recovery in alkali-burned rats, which had a protective effect on vision. It has been demonstrated that anti-vascular endothelial growth factor (VEGF) drugs could inhibit VEGF receptor activity and thus reduced the activity of the dopamine (DA) system and disrupted the balance between D1 and D2 receptors, and then the imbalanced DA system could cause prolongation of the eye axis and increased myopia. β-sitosterol can activate the KDR gene, which is highly associated with its function, and can activate the VEGF receptor encoded by KDR gene which can increase and adjust DA levels in the retina so as to achieve the purpose of treating myopia. In addition, studies have pointed out that the lack of ocular blood perfusion could exacerbate myopia, and retinal vessel diameter and blood perfusion were greater in patients with milder myopia.,43 At the same time, intraocular inflammatory factors could promote the exacerbation of myopia and lead to retinal lesions in patients. Existing pharmacological studies in Chinese medicine have demonstrated that the active constituents of DZP, such as β-sitosterol, umbelliferyl and isohesperidin, had anti-inflammatory and immunomodulatory effects as well as functions of vasodilating and improving blood circulation,–48 which may help to restore eye blood perfusion, alleviate eye verification and thus treat myopia, but the specific mechanisms remain to be clarified.Active constituents involved in the therapeutic effects of DZP on myopia mainly included quercetin, β-sitosterol, Diincarvilone A, methyl ferulate, and naringenin. Quercetin is a flavonol that can be processed into ophthalmic ointment preparations and has good efficacy in the treatment of metabolic cataract. As a signaling protein, it could act on the photoreceptor outer segments of the retina, and continuous chronic stimulation from STAT3 could cause retinal lesions and aggravate myopia. HSP90AA1 is a heat shock protein (HSP) and its functions are related to immunity, structural maintenance and repair. The expression of α-Crystallin, a major component of the lens which also belongs to HSP, can interact with degeneration proteins to inhibit degeneration protein aggregation and therefore maintain the size and transparency of the lens and can reduce the number of degeneration proteins to avoid the formation of severe myopia.The 6 important intersectional targets of DZP for myopia treatment were STAT3, PIK3CA, PIK3R1, MAPK1, MAPK3, and HSP90AA1. In recent years, STAT3 has gradually gained attention in the field of ophthalmology. One study found the increased expression of activated STAT3 in the retina of FDM guinea pigs, suggesting that STAT3 may be involved in the formation and development of myopia. The overexpressed VEGF can promote the excessive proliferation of retinal blood vessels, which will lead to the development of low myopia to high myopia, and even lead to retinal structural damage, macular degeneration, cataract, etc. According to the result of Figure  Therefore, DZP may increase ATP level by the “positive regulation of ATP metabolic process” pathway so as to treat myopia.The enrichment analyses of GO and KEGG found that the pathways closely related to DZP and myopia targets were “AGE-RAGE signaling pathway in diabetic complexes” and “positive regulation of ATP metabolic process” pathways. The AGE-RAGE signaling pathway can regulate the binding of advanced glycosylation end-products (AGE) to the receptor of advanced glycosylation end-products (RAGE), which can further activate Muller glial cells so as to promote VEGF secretion. In addition, Juhua can reduce platelet activation derivative TXA2 and platelet adhesion rate, which can repair abnormal platelet activation function. In one study, after the administration of asparagus decoction to SD rats, neuroactive enzyme levels in the brain tissue of SD rats increased and acetylcholine transferase levels decreased, showing a neuroactive role. The studies mentioned above may help to support our hypothesis that the effect of DZP in treating myopia is achieved through the “platelet activation” and “neuroactive ligand receptor interactions” pathways.The results of cross-target enrichment analyses by WGCNA and DEG analysis showed that the pathways closely related to myopia mainly involved “platelet activation” and “neuroactive ligand receptor interactions.” Studies have shown that asparagus, a constituent of DZP, had the effect of reducing platelet aggregation. Juhua can inhibit the activation and phagocytosis of macrophages, and asparagus can increase the proportions of CD4T cells and plasma cells. Based on the immune regulatory effects of the constituent ingredients of DZP and the immune infiltration results of myopia samples, it is predicted that DZP may play an active role in regulating myopia.Our study showed that the proportions of inactivated mast cells and M2 macrophages increased and the proportions of activated dendritic cells, activated mast cells, plasma cells, and CD4 T cells were lower in people with myopia. One study showed that after the administration of DZP to mice, the number of Langerhans dendritic cells in the skin as well as their number of dendrites increased.Although the downstream targets of key myopia genes predicted by WGCNA and DEG analysis were not among the predicted targets of DZP, the molecular docking results showed that MIP and LGSN could firmly bind to the active constituents of DZP. MIP could bind to Dincarvillone A with a binding energy of −36.233 kJ/mol, which was second only to the stability of PIK3R1 and Dincarvillone A in 40 docking groups. The results of molecular docking suggested that the active constituents of DZP could effectively and stably combine with the myopia core targets, which may provide supports for further research to confirm the therapeutic effect of DZP for treating myopia.This study revealed the potential mechanisms of DZP for myopia through diverse analyses and fastidious methodology. However, the results of immune infiltration and bioinformatic analyses cannot provide direct evidence for the efficacy of DZP on myopia. Randomized-controlled trials are required to evaluate its efficacy directly. Presently, there are no randomized controlled trials on DZP for myopia, so it is not available to provide systematic evaluation evidence. The active constituents identified by this study, quercetin and beta-sitosterol, are widespread polyphenols, known for nonspecific in silico effects and act as pan-assay interfering substances. Therefore, the conclusions of these 2 substances should be treated with caution, and whether they can play a role in the treatment of myopia also needs to be further verified by subsequent research. DZP can be used as an auxiliary treatment for the pathological damages of myopia, especially in the treatment of high myopia, because it can inhibit the progression of macular degeneration and cataract through the function of anti-VEGF, and its function of improving ATP level can alleviate the lengthening of ocular axis and the degeneration of eyeball caused by ciliary muscle spasm, thus preventing the occurrence of retinal detachment in high myopia. In addition, clinicians are suggested to properly adopt Dincarvilone A to treat myopia, because its strong interactions with other intersectional targets and high binding energies with the core targets of myopia. When treating myopia patients with diabetes, more attention can be paid to the regulation of AGE-RAGE pathway. Subsequent studies can further study the pharmacological effects of DZP on myopia and develop new drugs according to the targets and pathways revealed in this study. This study has proved the effects of DZP on the basis of bioinformatics and network analyses, which can provide reference for the follow-up studies to conduct randomized-controlled trials of DZP in the treatment of myopia.Based on the network analysis and bioinformatic analyses, this study provided a noninvasive and moderate treatment for myopia, filled the gap in the research of the mechanisms traditional Chinese medicine in treating myopia, and provided guidance for the future research direction on the pharmacological effects of DZP in treating myopia. DZP may exert its efficacy by regulating the DA system, balance between D1 and D2 receptors and the levels of VEGF and ATP in the eyes. Clinically, low or middle myopia generally does not cause great pathological damages to patients, but high myopia can bring a variety of pathological damages, such as macular degeneration, cataract, and retinal detachment, etc.In summary, based on the network analysis and bioinformatics analyses, this study suggested that the active constituents of DZP could act on intersectional targets: STAT3, PIK3CA, PIK3R1, MAPK1, MAPK3, and HSP90AA1, core targets of myopia: MIP and LGSN, “AGE-RAGE signaling” and “positive regulation of ATP metabolic process” pathways to treat myopia. DZP active constituents and myopia targets could bind stably and immune infiltration analysis showed the potential mechanisms for the efficacy of DZP.Conceptualization: Longkun Liu, Yan Zhao, Li Li, Zhaolan Liu.Data curation: Longkun Liu.Formal analysis: Longkun Liu, Li Li, Zhaolan Liu.Funding acquisition: Li Li, Zhaolan Liu.Investigation: Zhaolan Liu.Methodology: Longkun Liu, Yoann Birling, Yan Zhao, Wenxin Ma, Yang Tang, Yuxin Sun, Xuehui Wang, Mingkun Yu, Li Li, Zhaolan Liu.Software: Longkun Liu, Zhaolan Liu.Writing – original draft: Longkun Liu, Yoann Birling, Yan Zhao, Wenxin Ma, Yang Tang, Yuxin Sun, Xuehui Wang, Mingkun Yu, Hongsheng Bi, Jianping Liu, Li Li, Zhaolan Liu.lili_bidc@163.com).”This article was published with errors in the contact information for the corresponding author. The correct contact information is “*Correspondence: Li Li, Beijing Institute for Drug Control, NMPA Key Laboratory for Safety Research and Evaluation of Innovative Drugs, Beijing Key Laboratory of Analysis and Evaluation on Chinese Medicine, Beijing, China (