Research status of Bovis Calculus and relevant Chinese patent medicines based on bibliometric analysis / 中国中药杂志

With scarce resources, natural Bovis Calculus is expensive and hard to meet clinical demand. At the moment, four kinds of Bovis Calculus are available on the market: the natural product, in vitro cultured product, synthesized product, and the product formed in cow after manual intervention. In this study, papers on the four kinds of Bovis Calculus products and relevant Chinese patent medicines were searched from Web of Science, PubMed, and China National Knowledge Infrastructure(CNKI). CiteSpace, citexs AI, and CNKI were employed for bibliometric analysis and knowledge map analysis. On this basis, the status, trend, and focuses of research on Bovis Calculus and relevant Chinese patent medicines were summarized. The results suggested overall slow development in the research on Bovis Calculus and relevant Chinese patent medicines with three typical growth stages. It is consistent with the development of Bovis Calculus substitutes and the national policy for the development of traditional Chinese medicine. At the moment, the research on Bovis Calculus and relevant Chinese patent medicines has been on the rise. In recent years, there has been an explosion of research on them, particularly the quality control of Bovis Calculus and the Chinese patent medicines, the pharmacological efficacy of Chinese patent medicines, such as Angong Niuhuang Pills, and the comparison of the quality of various Bovis Calculus products. However, there is a paucity of research on the pharmacological efficacy and the mechanism of Bovis Calculus. This medicinal and the relevant Chinese patent medicines have been studied from diverse perspectives and China becomes outstanding in this research field. However, it is still necessary to reveal the chemical composition, pharmacological efficacy, and mechanism through multi-dimensional deep research.

Protective Effect of Glycyrrhizae Radix et Rhizoma-Granati Pericarpium Formula on Ultraviolet-induced Skin Damage / 中国实验方剂学杂志

Objective:To explore the potential synergistic protective mechanism of Glycyrrhizae Radix et Rhizoma-Granati Pericarpium formula compound by using the methods and tools of network pharmacology，and provide a basis for the modernization of traditional Chinese medicine（TCM） compounds and the discovery of new drugs. Method:Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） was used to obtain the active components of Glycyrrhizae Radix et Rhizoma-Granati Pericarpium formula and their corresponding targets. The obtained targets were input to the UniProt database to inquire the gene names corresponding to the targets. By searching the CTD database，Genecards database and OMIM database of disease-related websites，the anti-sunburn targets were obtained. The interaction of the active targets was analyzed with online STRING database to screen the predicted core targets. The gene ontolog（GO） gene function enrichment analysis and Kyoto encyclopedia of genes and genomes（KEGG） pathway enrichment analysis of the predictive targets were performed by using DAVID database. Cytoscape 3.6.1 software was used to make "drug-component-target" network diagram，"protein-protein interaction" network diagram and "component-target-pathway" network diagram. Online website Draw Venn Diagram was used to show the relationship between disease targets and drug predicted targets. R Studio software was used to draw the functional enrichment analysis diagram of GO gene and KEGG pathway. Molecular docking between the active ingredients and the core targets was performed using GOLD software. Result:The 16 active compounds were collected，such as liquiritin，glycyrrhizin，kaempferol and quercetin. The active components mainly acted on 5 core targets：protein kinase B1（AKT1），interleukin（IL）-6，vascular endothelial growth factor（VEGFA），tumor necrosis factor（TNF） and tumor suppressor gene （TP53） and played a role in anti-sunburn effect primarily through these pathways such as hepatitis B，pathways in cancer，toxoplasmosis，chagas disease（American trypanosomiasis），and TNF signaling pathway. Conclusion:Based on the method of network pharmacology，the present study has preliminarily explored the anti-sunburn targets and pathways of Glycyrrhizae Radix et Rhizoma-Granati Pericarpium formula，and further verified the characteristics of multi-component and multi-target treatment of diseases in TCM，so as to provide certain scientific ideas for the modernization research of Chinese herbal compound prescriptions.

Analysis of water-soluble nutrients in Lycium barbarum leaves and differences between different producing areas / 中国中药杂志

This study is to clarify the composition and content differences of water-soluble nutrients in Lycium barbarum leaves(LBLs) from different areas. The total polysaccharides, free monosaccharides and oligosaccharides, nucleosides and amino acids in 35 batches of LBLs were analyzed with use of spectrophotometry, HPLC-ELSD and UPLC-MS/MS. The results showed that LBLs contained abundant polysaccharides, fructose, glucose, sucrose and maltose, with an average contents of 39.07, 12.69, 8.99, 17.44, 8.32 mg·g~(-1), respectively. Besides, eight nucleosides and twelve amino acids were detected in LBLs, and their average total contents were 54.95, 336.9 μg·g~(-1). Principal component analysis(PCA) and partial least squares discrimination analysis(PLS-DA) of carbohydrate, nucleoside and amino acid showed that the water-soluble nutrients of the samples from Qinghai Province were significantly different from those from other areas mainly in asparagine, proline, glutamine, sucrose, adenine and guanosine. In this study, the compositions and contents of water-soluble nutrients in LBLs were preliminarily clarified, which provided basis for further development and utilization of LBLs resoures.

Anti-inflammatory Mechanism of Epimedii Folium in Treatment of Depression Based on Network Pharmacology and Molecular Docking / 中国实验方剂学杂志

Objective:To predict the anti-inflammatory targets and relevant signaling pathways of Epimedii Folium in the treatment of depression by network pharmacology，in order to explore the potential mechanism of its anti-depression effect. Method:The active constituents of Epimedii Folium were collected and screened out through the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform（TCMSP） database. PharmMapper server and TCMSP database were used to predict and screen out protein targets. OMIM database，CTD database and GeneCards database were used to screen out relevant targets and anti-inflammatory targets of depression. Enrichment analysis of the gene ontology （GO） function and Kyoto Encyclopedia of genes and genomes（KEGG） signaling pathway for the key anti-inflammatory targets of Epimedii Folium were carried out by DAVID database. Cytoscape 3.6.0 was used to construct the network diagram of "active component-action target-signal pathway" of Epimedii Folium and analyze the topological structure of the network. GOLD molecular docking software was used to verify the results of active components and key anti-inflammatory targets. Result:A total of 12 active components，30 targets and 5 key anti-inflammatory targets of Epimedii Folium were screened out， 65 biological processes，4 cell components and 1 molecular function were enriched with GO function， and 41 KEGG pathways were enriched and analyzed，including 9 inflammation-related signaling pathways. Molecular docking verified that icariin and key anti-inflammatory targets could form the optimal binding structure. Conclusion:The study preliminarily reveals the molecular mechanism of Epimedii Folium on depression through its anti-inflammatory target and its relevant signaling pathway network，so as to provide a basis for further study on the antidepressant effect of Epimedii Folium.

Molecular Cloning and Sequence Analysis of GgUGE Gene in Glycyrrhiza glabra / 中国实验方剂学杂志

Objective::To clone the cDNA sequence of UDP-glucose 4-epimerase (UGE) in Glycyrrhiza glabra and analyze its sequence, so as to explore the potential relationship between the UGE gene and the molecular regulatory mechanisms of glycyrrhizic acid biosynthesis. Method::The cDNA sequence of UGE was cloned from the root of G. glabra by reverse transcription polymerase chain reaction (RT-PCR), then sequenced and analyzed by bioinformatics software. Results::A GgUGE cDNA sequence with the full length of 1 121 bp was obtained. The open reading fame (ORF) of GgUGE was 1 053 bp, encoding 350 amino acid residues. The GgUGE cDNA sequence was submitted to GenBank, and the accession No. was MK638908. Sequence analysis showed that GgUGE was an unstable hydrophilic protein, its average relative molecular weight was 39.02 kDa, and isoelectric point was 6.13. It contained no signal peptides or transmembrane domains. Its secondary structure mainly constituted of α-helix and had a conversed domain of UDP-glucose 4-epimerase superfamily. The homologoue analysis showed that the cDNA and amino acid sequences of GgUGE had the closest evolutionary relationship to Leguminosae and relatively distant evolutionary relationship to Salicaceae. Conclusion::In this study, GgUGE cDNA sequence is successfully cloned from G. glabra for the very first time, which will provide reference for studying the function of GgUGE and explaining the molecular regulatory mechanisms of glycyrrhizic acid biosynthesis in G. glabra.

Regionalization of production of medicinal and edible fruit of Lycium barbarum associated with ecological factors and chemical constituents / 药学学报

The fruit of Lycium barbarum L. (FLB) is a food and medicinal herb. Identifying suitable production regions for this plant would be beneficial to its cultivation and production. In this study, the Maxent model was used to identify ecologically suitable regions for the growth of L. barbarum L. In addition, based on its chemical composition, the suitable regions for production were identified by literature analysis and chemometrics. The results show that suitable regions for L. barbarum L. culture are mainly distributed in the northwest of China; suitable regions for the production of medicinal FLB were mainly concentrated in the district of Ningxia, Baiyin, Jiuquan and Zhangye of Gansu, and parts district in west of Inner Mongolia. All are the traditional production regions for FLB, which is consistent with the good quality of FLB produced in Ganzhou in ancient times, and the genuine medicinal materials of FLB produced in Zhongning of Ningxia today. The suitable regions for edible FLB were mainly distributed in northwest of Qinghai, Jiuquan and Zhangye of Gansu, as well as Aksu and Kizi sukirgiz of Xinjiang. The fruit type index of FLB in these regions is large, and the content of fructose and glucose in the fruit is high, which satisfies the edible commodity property. The study results lay a foundation for realizing the regional distribution and development of L. barbarum based on its different uses.

Comparative analysis of twenty-five compounds in different parts of Astragalus membranaceus var. mongholicus and Astragalus membranaceus by UPLC-MS/MS / 药物分析学报

As a traditional Chinese medicine, the root of Astragalus membranaceus var. mongholicus (AMM) or A. membranaceus (AM) has been widely used in China and other Asian countries for thousands of years. Till now, the flavonoids, phenolic acids and saponins are considered as the main active components contributing to their therapeutic effect in these plants. In order to clarify the distribution and contents of these compounds in different organs of these plants, a rapid and sensitive analytical method for simultaneous determination of 25 active compounds including seven types (i.e. dihydroflavones, iso-flavane, isoflavones, flavones, pterocarpans, phenolic acid and saponins) within 10 min was established using ultra-pressure liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS). Then, the established method was fully validated and successfully applied to the determination of the contents of these analytes in different parts (root, rhizome, stem, leaf and flower) of AMM and AM. The results indicated that the contents of the same type of compounds in two different species plants were significantly different. Moreover, the obvious differences were also found for the distribution and con-tents of different type of compounds in five organs of the same species. The present study could provide necessary information for the rational development and utilization of AMM and AM resource.

Umbelliferone ameliorates renal function in diabetic nephropathy rats through regulating inflammation and TLR/NF-κB pathway / 中国天然药物

Diabetic nephropathy (DN) is a leading cause of renal failure, contributing to severe morbidity and mortality in diabetic patients. Umbelliferae (Umb) has been well characterized to exert protective effects in diabetes. However, the action and mechanism of Umb in DN remains unclear. In this work, we studied the effect of Umb in a streptozotocin (STZ)-induced DN rat model and explore its underlying mechanism. DN rats were treated withUmb (20, 40 mg·kg) orirbesartan (15 mg·kg) for 4 weeks. Levels of serum glucose, insulin, blood uric acid, creatinine, triglycerides (TG) and total cholesterol (TC) were measured bycommercial assay kits, respectively. Histopathological changes andinflammatory cytokine levels including IL-6, IL-1β and TNF-α in the kidney were also evaluated. Alterations in the expression of podocin, CD2AP and TLR/NF-κB were assessed by western blotting. Our results showed that Umb reduced renal injury in DN rat model, as evidenced by the decrease in blood glucose, 24 h urinary protein, serum creatinine, and blood uric acid. Umb also significantly ameliorated the renal histopathological alteration, and down-regulated the expression of epithelial-to-mesenchymal transition-related molecular markers podocin and CD2AP. Moreover, Umb inhibited TLR2, TLR4, MyD88 expressions, NF-κB activation and considerably reduced levels of other downstream inflammatory molecules (TNF-α, IL-6, IL-1β). These findings indicated that Umb improved renal function through regulating inflammation and TLR/NF-κB pathway, suggesting the potential efficacy of Umb in DN treatment.

Quantitative Analysis of Index Components in Glycyrrhizae Radix et Khizoma by Near Infrared Spectroscopy Based on R Software / 中国实验方剂学杂志

Objective: To establish a better near infrared quantitative model for quality control of Glycyrrhizae Radix et Rhizoma of components (moisture,total ash,liquiritin and glycyrrhizic acid) in liquorice,in order to realize rapid detection.Method: The contents of moisture,total ash,liquiritin and glycyrrhizic acid were determined in 97 samples based on the methods set forth in Chinese Pharmacopoeia.Meanwhile,the near infrared spectrum was scanned using near infrared spectroscope.R software was used to screen out the spectral pretreatment and build the quantitative models.Result: The optimum spectral pretreatment method for establishing the near infrared quantitative model of moisture and liquiritin was the first order derivative.For moisture,the correlation coefficients of test and validation were 0.930 0 and 0.929 9,and the root mean square errors were 0.243 2 and 0.203 8,respectively.For liquiritin,the correlation coefficients of test and validation were 0.930 3 and 0.907 6,and the root mean square errors were 0.093 9 and 0.128 9,respectively.The optimum spectral pretreatment method for establishing the near infrared quantitative model of total ash was MSC.The correlation coefficients of test and validation were 0.926 5 and 0.917 7,and the root mean square errors were 0.109 6 and 0.103 7,respectively.The optimum spectral pretreatment method for establishing the near infrared quantitative model of glycyrrhizic acid was SNV.The correlation coefficients of test and validation were 0.918 1 and 0.915 7,and the root mean square errors were 0.274 8 and 0.236 0,respectively.Conclusion: In this study,a better near infrared quantitative models for quality control of components of Glycyrrhizae Radix et Rhizoma were established,with a high accuracy,which laid a foundation for rapid detection of the components in Glycyrrhizae Radix et Rhizoma.

Network pharmacology of flavonoids in Sophora alopecuroides / 中国中药杂志

The aim of this paper was to investigate the potential pharmacological effect of flavonoids in Sophora alopecuroides by network pharmacology. This study predicted the potential targets of 11 flavonoids of S. alopecuroides with help of reversed pharmacophore matching target recognition service platform (PharmMapper). The pathway information was acquired from DAVID and KEGG databases. Cytoscape software was used to construct the "ingredient-target-pathway" network of flavonoids active components of S. alopecuroides. The flavonoids active components of S. alopecuroides play anti-inflammatory, blood sugar regulating and other pharmacological effects by regulating 62 targets (such as INSR，KDR，MET) and intervening 44 pathways, such as B cell receptor signaling pathway, insulin signaling pathway, neurotrophin signaling pathway, and T cell receptor signaling pathway. In this study, the mechanism of "muti components-multitargets-multiple pathway" of flavonoids was studied. It reflects the multi-components, multi-targets and multiple pathway features of traditional Chinese medicine. Meanwhile, it provides a scientific basis for the elucidation the mechanism of S. alopecuroides as a medicine, and the development and utilization resources of S. alopecuroides.

Research on network pharmacology of alkaloids in Sophora alopecuroides / 中国中药杂志

It was aimed at exploring the potential pharmacological effects of alkaloids in Sophora alopecuroides by means of network pharmacology in this study. The main alkaloids in S. alopecuroides were collected for analysis of drug properties, prediction of potential targets and screening of signaling pathways. DAVID analysis tool combined with KEGG database was used to annotate and analyze the signaling pathway. The alkaloids-targets-signaling pathways network was built through Cytoscape software. Results showed that 17 alkaloids in S. alopecuroides involved 49 targets (170 times in all) and 22 important signaling pathways. Three nodes in model of network pharmacology were cross-linked, and the metabolic pathways were coordinated and regulated by each other. It indicated that alkaloids in S. alopecuroides may have therapeutic effect on diseases of cancer, metabolic disorder, endocrine system, digestive system, nervous system and so on.

Comparative identification studies on wild and cultivated Glycyrrhiza uralensis produced Ningxia based on index components and near-infrared spectroscopy / 中国中药杂志

This study is to construct a rapid and effective method for identification of wild and cultivated Glycyrrhiza uralensis (hereinafter referred to as Glycyrrhizae Radix et Rhizoma) from Ningxia by comparison of the difference in chromatography identification based on index components and near-infrared spectroscopy identification. HPLC and UV methods were used to determine the content of liquiritin, glycyrrhizate and total flavonoids for 9 wild Glycyrrhizae Radix et Rhizoma and 14 cultivated Glycyrrhizae Radix et Rhizoma samples,and the near-infrared spectroscopy was also,collected. The results illustrated that the chromatography identification based on index components could not identify wild and cultivated Glycyrrhizae Radix et Rhizoma from Ningxia, while near-infrared spectroscopy could quickly and effectively achieve it. It provides an effective method for the growth pattern identification and application of Glycyrrhizae Radix et Rhizoma.

Effects of different drying methods on content of bioactive component and antioxidant activity in Lycium ruthenicum / 中国中药杂志

To compare the appearances, tastes, contents of bioactive components and antioxidant activity of Lyceum ruthenicum under different drying methods, so as to direct its production practice. The folin-phenol colorimetric method, UV, extinction coefficient method and DPPH, as well as fluorescence recovery after photobleaching (FRAP) method to determine the contents of polyphenols, proanthocyanidins, total anthocyanin and antioxidant activity under different drying methods： vacuum freeze drying, low-temperature oven drying and air drying for L. ruthenicum. The results showed that the drying methods had certain effects on its appearances, tastes, contents of bioactive components and antioxidant activity. The appearances and tastes were best after the L. ruthenicum was dried by vacuum freeze drying, with significantly lower moisture than air drying method. The contents of total polyphenols, anthocyanin and proanthocyanidins were highest by air-drying but lowest by low temperature oven drying in L. ruthenicum. The scavenging ability to DPPH was strongest by freeze-drying and lowest by low temperature oven drying, while the antioxidant activity was strongest by air-drying in the FRAT method. In addition, the appearances and tastes were poor in air drying, with higher moisture but highest contents of the three bioactive components. Therefore, the drying methods for L. ruthenicum shall be comprehensively considered.

Comparative studies on HPLC fingerprint and near-infrared spectra of cultivated and reference crude Scutellaria baicalensis / 中国中药杂志

Scutellaria baicalensis is a common and important medicinal plant in China, facing with reducing sharply in wild resources. To meet the needs in Chinese herbwouls medicine market and clinical application, S. baicalensis has been widely cultivated in Ningxia, Hebei, Shanxi, and Gansu et al. HPLC finger-print and near-infrared were studied in the research to evaluate quality difference of S. baicalensis in four districts. The results showed that the similarity of HPLC finger-print of 12 cultivated S. baicalensis and reference crude herb is more than 0.961, and the other is more than 0.983. On the other hand, paired sample t-test indicated there has no significant difference between the common peaks' area of 12 cultivated S. baicalensis and reference crude herb. It was verified that 12 cultivated S. baicalensis has highly consistency with reference crude herb. On the basis of chromatographic finger-print and near-infrared spectrum, the study applied paired sample t-test to verify analysis results, which could avoid erroneous judgment induced by indefinite threshold values in the similarity of chromatographic finger-print and provide reliable basis for the analysis results. Meanwhile, it also provides a new idea for improving the quality control method of Chinese medicinal materials by comparative study about two comprehensive detection means.

Production regionalization study of Lycii Fructus / 中国中药杂志

The distribution information of Lycii Fructus was collected by interview investigation and field survey, and 46 related environmental factors were collected, some kinds of functional chemical constituents the of Lycii Fructus were analyzed. Integrated climate, topography and other related ecological factors, the habitat suitability study was conducted based on Arc geographic information system(ArcGIS),and maximum entropy model. The AUC of ROC curve was both above 0.95, indicating that the predictive results with the maximum model were highly precise. The results showed that 5 major ecological factors had obvious influence on ecology suitability distributions of Lycii Fructus, including soil pH, soil subclass, vegetation type and in August the average temperature et al. It is suitable for the living habits of the Lycii Fructus, dry, cool weather, more hardy, drought-resistant, alkali soil, which is suitable for distribution in the northern temperate plains. In addition, the ecological suitability regionalization based on the chemical constituents of Lycii Fructus also provides a new suitable distribution area other than the traditional distribution area, which provides a scientific basis for the reasonable introduction of Lycii Fructus.

Production regionalization study of Glycyrrhiza uralensis / 中国中药杂志

The distribution information of Glycyrrhiza uralensis was collected by interview investigation and field survey, and 46 related environmental factors were collected, some kinds of functional chemical constituents of G.uralensis were analyzed. Integrated climate, topography and other related ecological factors, the habitat suitability study was conducted based on Arc geographic information system(ArcGIS),and maximum entropy model. The AUC of ROC curve was both above 0.95, indicating that the predictive results with the maximum model were highly precise. The results showed that 5 major ecological factors have obvious influence on ecology suitability distributions of G. uralensis, including July average temperature, soil sub category, Dec precipitation, vegetation types and standard deviation of seasonal variation in temperature, et al. It is suitable for the living habits of the G. uralensis, adequate light, low rainfall, summer heat and large temperature difference between day and night, which is suitable for distribution in the northern temperate plains and mountains. In addition, the ecological suitability regionalization based on the chemical constituents of G.uralensis also provides a new suitable distribution area other than the traditional distribution area, which provides a scientific basis for the reasonable introduction of G.uralensis.

Study on triterpenes from of Ligularia xanthotricha / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the chemical constituents of Ligularia xanthotricha.</p><p><b>METHOD</b>Silica gel column chromatography and preparative TLC were employed for the isolation and purification. The structures were identified on the basis of spectral data (IR, EI-MS, 1H-NMR, 13C-NMR, DEPT) and chemical evidence.</p><p><b>RESULT</b>Seven compounds were isolated and identified as follows: lupeol (1), lupeol palmitate (2), 3, 28-dihydroxyl-lupeol (3), betulinic acid (4), taraxasterol (5), taraxasteryl palmitat (6) and taraxasteryl acetate(7).</p><p><b>CONCLUSION</b>All the compounds were isolated from this plant for the first time.</p>

Study on chemical constiuents from Ligularia intermedia of shanxi / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents of Ligularia intermedia of Shanxi.</p><p><b>METHOD</b>The compounds were isolated by column chromatography on silica gel and preparative TLC. The structures were identified by IR, MS, 1D/2DNMR spectral data and X-ray single crystal diffraction and other methods1.</p><p><b>RESULT</b>Nine compound were isolated and identified as 8beta-hydroxyeremophil-7(11)-ene-12, 8alpha(4beta, 6alpha)-diolide (1), 8beta-methoxyeremophil-7(11)-ene-12, 8alpha(4beta, 6alpha)-diolide (2), petasin (3), isopetasin (4), liguhodgsonal (5), ligudentatol (6), ligujapone (7), lupeol (8) and lupeol palmitate (9).</p><p><b>CONCLUSION</b>Compounds 2, 3, 4, 6, 7 and 9 were isolated from the plant for the first time.</p>

Studies on flavonoid constituents from herbs of Artemisia ordosica II / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents of herbs of Artemisia ordosica.</p><p><b>METHOD</b>The chemical constituents were isolated and repeatedly purified on silica gel column and the structures were elucidated by the NMR spectra and physicochemical properties.</p><p><b>RESULT</b>Eight flavones were obtained and they were identified as isosakuranetin, 7, 4'-dimethylaro madendrin, acacetin, cirsimaritin, rhamnetin, eupatolin, 5, 7, 2', 4'-tetrahydroxy-6, 5'-dimethoxyflavone, hyperoside.</p><p><b>CONCLUSION</b>All the compounds were obtained from herbs of A. ordosica for the first time.</p>

Natural Anti-infectious Molecule:Bactericidal/permeability- increasing Protein / 中国生物工程杂志

Endotoxins are lipopolysaccharides ( LPS) , major component of out wall in Gram-negative bacteria. Often they are considered as "prime criminals" of triggering systemic inflaming reaction such as sepsis, bacteremia and so on. It was found in recent years that bactericidal/permeability-increasing protein, a 55kDa member of lipid-related protein family, was a kind of natural molecule of anti-infection and it has special endotoxin-neutralising activity and antibacterial activity. Comprehensive phase II/III clinical trials demonstrated the feasibility and safety of recombinant BPI. So pharmaceutical corporations are attracted to try to apply it to therapy.