ObjectiveTo investigate the effects of exogenous nitric oxide （NO） on the accumulation and quality formation mechanism of flavonoids in Scutellariae Radix. MethodsFresh roots of Scutellaria baicalensis were treated with sodium nitroprusside （SNP） solutions at concentrations of 0.0， 7.5， and 20 mmol·L^-1， respectively. Kits and supporting reaction systems were used to determine the following indicators of samples in each group， including （1） reactive oxygen species： changes in the content of nitric oxide （NO）， superoxide anion （O2-·）， and hydrogen peroxide （H₂O₂）， （2） lipid oxidation products： changes in the content of malondialdehyde （MDA）， （3） antioxidant enzymes： changes in the activity of superoxide dismutase （SOD）， catalase （CAT）， and peroxidase （POD）， and （4） metabolic key enzymes and metabolic intermediates： Changes in the activity of phenylalanine ammonia-lyase （PAL） and the content of 1，3-diphosphoglycerate （1，3-DPG）. High-performance liquid chromatography （HPLC） was used to determine the changes in the content of secondary metabolites in flavonoids such as baicalin， wogonoside， baicalein， and wogonin in each group. ResultsCompared with that on day 0， during the treatment with 20 mmol·L^-1 SNP solution， the content of NO， O2-·， H₂O₂， and MDA in fresh roots of Scutellaria baicalensis increased significantly （P<0.01）， by 138.7%， 75.7%， 292.9%， and 140.3% respectively. The activity of SOD， CAT， and POD increased by 100.7%， 41.4%， and 70.4% respectively （P<0.01）， and that of PAL increased by 111.1% （P<0.01）. The content of the secondary metabolites baicalin and wogonoside remained basically unchanged， while that of baicalein and wogonin increased by 401.3% and 457.9% respectively （P<0.01）. The content of 1，3-DPG increased by 165.0% （P<0.01）. In the 7.5 mmol·L^-1 SNP solution treatment group， the stress degree was relatively mild. Compared with that on day 0， the content of NO， O2-·， H₂O₂， and MDA increased by 76.2%， 57.7%， 221.4%， and 99.7% respectively （P<0.01）. The activity of antioxidant enzymes SOD， CAT， and POD increased by 79.5%， 55.6%， and 51.9% respectively （P<0.01）， and that of PAL increased by 69.0% （P<0.01）. The content of baicalein and wogonin increased by 204.7% and 272.2% respectively （P<0.01）， and that of 1，3-DPG increased by 128.0% （P<0.01）. The 20 mmol·L^-1 SNP group showed better performance than the 7.5 mmol·L^-1 SNP group. ConclusionSNP can simulate the physiological state of Scutellaria baicalensis under stress conditions， activate the antioxidant protection mechanism， promote the biosynthesis of flavonoids in Scutellariae Radix， and significantly improve the quality of medicinal materials in Scutellariae Radix.

ObjectiveTo investigate the therapeutic mechanism of Xuefu Zhuyutang （XFZYT） for metabolic-associated fatty liver disease （MAFLD） through integrated network pharmacology and animal experiments. MethodsNetwork pharmacology was utilized to predict the core components， key therapeutic targets， and signaling pathways of XFZYT in the treatment of MAFLD. For animal experiments， a rat model of MAFLD was established by feeding a high-cholesterol diet for 4 weeks. Intervention was then administered with low-dose （2 g·kg^-1） and high-dose （4 g·kg^-1） XFZYT for 2 weeks. Biochemical assays were performed to measure the serum levels of aspartate aminotransferase （AST）， alanine aminotransferase （ALT）， total cholesterol （TC）， triglycerides （TG）， high-density lipoprotein （HDL）， and low-density lipoprotein （LDL）. In addition， the activities of superoxide dismutase （SOD） and catalase （CAT） and levels of malondialdehyde （MDA） and glutathione （GSH） in the serum were measured. The same way was adopted to measure the levels of TC and TG in the liver tissue. Enzyme-linked immunosorbent assay （ELISA） was employed to quantify the serum levels of interleukin （IL）-6， IL-1β， and tumor necrosis factor-alpha （TNF-α）. Histopathological evaluations included hematoxylin and eosin （HE） staining for liver tissue morphology， Oil Red O staining for lipid deposition， and dihydroethidium （DHE） probe staining for reactive oxygen species （ROS） levels. Western blot analysis was conducted to assess the protein levels of AMP-activated protein kinase （AMPK）， phosphorylated （p）-AMPK， nuclear factor erythroid 2-related factor 2 （Nrf2）， heme oxygenase-1 （HO-1）， nuclear factor-kappa B （NF-κB）， and p-NF-κB in the liver tissue. Untargeted metabolomics analysis of the serum was performed by liquid chromatography-tandem mass spectrometry （LC-MS/MS）. ResultsNetwork pharmacology analysis predicted 155 potential targets of XFZYT for MAFLD treatment， with core targets including signal transducer and activator of transcription 3 （STAT3）， protein kinase B1 （Akt1）， TNF， and IL-6. Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment primarily implicated the AMPK signaling pathway. Animal experiments demonstrated that compared with the normal group， the model group exhibited dyslipidemia， hepatic function impairment， pronounced hepatic lipid deposition， and inflammatory manifestations， with elevated serum levels of AST， ALT， TC， TG， LDL， and MDA （P<0.05）， reduced HDL and GSH levels plus decreased SOD and CAT activities （P<0.05）， downregulated protein levels of Nrf2， HO-1， and p-AMPK （P<0.05）， and upregulated protein level of p-NF-κB （P<0.05） in the liver tissue. Compared with the model group， XFZYT intervention groups showed significant amelioration of dyslipidemia and hepatic function impairment， markedly reduced hepatic lipid deposition and inflammatory cell infiltration， decreased serum levels of AST， ALT， TC， TG， LDL， and MDA （P<0.05）， increased HDL and GSH levels plus enhanced SOD and CAT activities （P<0.05）， upregulated protein levels of Nrf2， HO-1， and p-AMPK （P<0.05）， and downregulated protein level of p-NF-κB （P<0.05）. Serum metabolomics revealed 511 differentially expressed metabolites （231 upregulated and 280 downregulated） between normal and model groups， while XFZYT groups versus model group showed 94 differential metabolites （51 upregulated and 43 downregulated）. Among them， 11 metabolites displayed the most significant alterations， with enriched pathways including glycerolipid metabolism， cholesterol metabolism， and insulin resistance， multiple of which demonstrated AMPK association. ConclusionXFZYT alleviates MAFLD by regulating the AMPK signaling pathway and associated metabolic networks.

AIM:To identify surrogate clinical bio-markers for profiling the eosinophilic status of an individual patient over induced sputum analysis.METHODS:We conducted a cross-sectional study on 100 asthmatic patients whose induced sputum was successfully collected.Subjects were further classified into either EA or NEA based on whether the percentage of sputum eosinophil count(SEC％)was≥3％.Demographic and clinical data were col-lected,including basic information,routine blood tests,lung function tests,bronchodilator reversibili-ty tests,fractional exhaled nitric oxide(FeNO),the Asthma Control Test(ACT)and the Asthma Control Questionnaire(ACQ).All variables significantly asso-ciated with EA were candidates for multivariate lo-gistic regression analysis.A scoring system present-ed as a nomogram for the prediction of EA was de-veloped.RESULTS:In the univariate analysis,com-pared with NEA subjects,those with EA were of older age and had worse asthma control and lung function in addition to higher values of blood eosin-ophils,serum IgE and FeNO.Multivariable logistic regression analysis revealed that age,FeNOx serum IgE and blood eosinophil count(BEC)were identi-fied as independent risk factors for eosinophilic asthma,which were all included in the nomogram.CONCLUSION:A combination assessment including age,FeNOx serum IgE and BEC could be applicable to clinicians in identifying eosinophilic asthma and is easier,faster,more inexpensive and more readily available than the induced sputum test.

Objective:To conduct a survey on the influence of personality trait on depres-sive symptom and obesity in patients before bariatric and metabolic surgery.Methods:The retros-pective cross-sectional investigation study was conducted. Patients who underwent bariatric and metabolic surgery in The Third Xiangya Hospital of Central South University and The First Affiliated Hospital of Ji′nan University from January 1 to June 1,2024 were recruited. The survey was conducted in electronic format using the general information survey questionnaire, the patient health question-naire-9 (PHQ-9), and the Chinese big five personality inventory brief version (CBF-PI-B). Observa-tion indicators: (1) results of the questionnaire; (2) general data of patient; (3) personality trait scores for patient with different subtypes of obesity; (4) depression symptom scores for patient with different subtypes of obesity; (5) correlation analysis for patient personality trait, depressive symptom and body mass index (BMI). Comparison of measurement data with normal distribution between groups was conducted using the independent t test, and one-way analysis of variance was used for comparison among multiple groups. LSD test was used for pairwise comparison. Comparison of measurement data with skewed distribution among multiple groups was conducted using the Kruskal-Wallis H test, and Dunn Bonferroni test was used for pairwise comparison. The Spearman correlation analysis and Pearson correlation analysis were used for correlation analysis, and the mediation effect analysis was conducted using the Process 4.0 model and the Bootstrap test. Results:(1)Results of the questionnaire. A total of 146 questionnaires were distributed and collected, including 83 questionnaires from The Third Xiangya Hospital of Central South University and 63 questionnaires from The First Affiliated Hospital of Ji′nan University. Of 146 questionnaires, 137 questionnaires were valid, yielding an effective response rate of 93.836%(137/146). (2)General data of patient. Of the 137 patients who completed valid questionnaires, 52 were male with BMI of (41±5)kg/m 2, 85 were female with BMI of (38±6)kg/m 2, showing a significant difference between them ( t=2.586, P<0.05). There were 40 patients aged ≤25 years, with BMI of (42±5)kg/m 2, 28 patients aged 26-30 years, with BMI of (39±5)kg/m 2, 49 patients aged 31-40 years, with BMI of (38±6)kg/m 2, 14 patients aged 41-50 years, with BMI of (38±7)kg/m 2, 6 patients aged >50 years, with BMI of (37±3)kg/m 2. There were significant differences in BMI of patients of different age groups ( F=2.697, P<0.05). (3)Persona-lity trait scores for patient with different subtypes of obesity. Of the 137 patients who completed valid questionnaires, there were 15 patients of mild obesity, with personality trait neuroticism score of 19.0(14.0,24.0), personality trait strictness score of 32.5±5.2, personality trait extroversion score of 30.0(24.0,40.0), 36 patients of moderate obesity, with personality trait neuro-ticism score of 26.5(22.3,28.8), personality trait strictness score of 32.6±5.9, personality trait extro-version score of 34.0(26.3,39.0), 81 patients of severe obesity, with personality trait neuroticism score of 35.0(29.5,41.0), personality trait strictness score of 26.8±7.4, personality trait extroversion score of 28.0(20.5,35.0), 5 patients of extreme obesity, with personality trait neuroticism score of 28.0(26.5,44.0), personality trait strictness score of 19.6±3.4, personality trait extroversion score of 22.0(18.5,25.0). There were significant differences in personality trait neuroticism, strictness and extroversion among patients of different subtypes of obesity ( H=50.316, F=10.774, H=14.446, P<0.05). (4)Depression symptom scores for patient with different subtypes of obesity. Of the 137 pati-ents who completed valid questionnaires, the depressive symptom score were 4.0(2.0,9.0) for 15 pati-ents of mild obesity, 5.0(3.0,7.0) for 36 patients of moderate obesity, 13.0(8.5,17.0) for 81 patients of severe obesity, and 18.0(9.5,22.0) for 5 patients of extreme obesity, respectively, showing a significant difference among them ( H=50.129, P<0.05). (5) Correlation analysis for patient personality trait, depressive symptom and BMI. Results of Spearman correlation analysis showed that personality trait neuroticism were positively correlated with depressive symptoms and BMI ( r=0.780, 0.557, P<0.05), personality trait strictness was negatively correlated with depressive symptoms ( r=-0.523, P<0.05), personality trait extroversion were negatively correlated with depressive symptoms and BMI ( r=-0.448, -0.323, P<0.05), and depressive symptoms was positively correlated with BMI ( r=0.568, P<0.05). Results of Pearson correlation analysis showed that personality trait strictness was negatively correlated with BMI ( r=-0.489, P<0.05). Conclusion:Preoperative personality trait of patients undergoing bariatric and metabolic surgery can influence their depressive symptom and BMI.

Background Recent advances in understanding the toxic effects of inorganic arsenic have revealed that arsenic exposure impacts multiple endocrine organs, thereby altering their functions. However, the mechanisms underlying arsenic-induced thyroid injury remain unclear. Objective To investigate the mechanisms by which sodium arsenite (NaAsO₂) affects the proliferation and apoptosis of normal thyroid cells (Nthy-ori3-1) through the estrogen receptor (ER)-phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway. Methods Nthy-ori3-1 cells were cultured in vitro and divided into the following groups: a control group (complete medium without drugs, 0 μmol·L⁻¹), and NaAsO₂-treated groups at 1, 2, and 4 μmol·L⁻¹. Additionally, 1 μmol·L⁻¹ of the ER inhibitor ICI182780 was used to intervene in the NaAsO₂ exposure groups, resulting in the following combinations: 1 μmol·L⁻¹ NaAsO₂ + ICI182780, 2 μmol·L⁻¹ NaAsO₂ + ICI182780, and 4 μmol·L⁻¹ NaAsO₂ + ICI182780. The median lethal concentration of NaAsO₂ was determined using cell viability assay. Cell viability was assessed at 24, 36, and 48 h using Cell Counting Kit-8 (CCK-8) assay. Colony formation ability was evaluated via plate cloning assay. Apoptosis was detected using Hoechst 33342 staining. Protein and mRNA expression levels of ERα, ERβ, c-MYC, Bax, Bcl-2, PI3K, p-PI3K, AKT, and p-AKT were measured using Western blot (WB) and real-time quantitative PCR (qRT-PCR), respectively. Results The median lethal concentration of NaAsO₂ was determined to be 4.034 μmol·L⁻¹. CCK-8 assay at 24, 36, and 48 h revealed that, compared with the control group, the 1, 2, and 4 μmol·L⁻¹ NaAsO₂ groups significantly inhibited Nthy-ori3-1 cell proliferation (P < 0.001). The plate cloning assays demonstrated a concentration-dependent reduction in colony formation ability (P < 0.001). Following the ICI182780 intervention, the cell viability and colony formation ability in the 1, 2, and 4 μmol·L⁻¹ NaAsO₂ groups were significantly restored compared with the corresponding NaAsO₂-only groups (P < 0.001, P < 0.01). The Hoechst 33342 staining indicated that compared with the control group, the nuclear staining intensity and apoptosis levels in the 1, 2, and 4 μmol·L⁻¹ NaAsO₂ groups increased in a concentration-dependent manner (P < 0.001). However, the ICI182780 intervention reduced the apoptosis levels in the NaAsO₂-treated groups compared with their NaAsO₂-only counterparts (P < 0.001). The WB analysis showed that, compared with the control group, the protein expression of ERα, ERβ, c-MYC, Bcl-2, p-PI3K, and p-AKT in the 1, 2, and 4 μmol·L⁻¹ NaAsO₂ groups decreased manner (P < 0.001, P < 0.01), while the Bax expression increased in a concentration-dependent (P < 0.001); the PI3K and AKT protein levels showed no significant differences (P > 0.05). In the ICI182780-treated NaAsO₂ groups, the ERα, ERβ, c-MYC, Bcl-2, p-PI3K, and p-AKT protein expression increased (P < 0.001, P < 0.01), the Bax expression decreased (P < 0.001), and the PI3K and AKT levels remained unchanged (P > 0.05) compared with the corresponding NaAsO₂-only groups. The mRNA expression patterns of ERα, ERβ, c-MYC, Bcl-2, and Bax were consistent with the WB results. Conclusion NaAsO₂ inhibits proliferation and promotes apoptosis in Nthy-ori3-1 cells in a dose-dependent manner. The underlying mechanism likely involves NaAsO₂-mediated suppression of the ER-PI3K/AKT signaling pathway, which subsequently regulates downstream proliferation- and apoptosis-related genes.

Non-coding RNAs(ncRNAs),encompassing microRNAs(miRNAs),long non-coding RNAs(lncRNAs),and circular RNAs(circRNAs),have emerged as critical regulators of gene expression and cellular function.In alcohol-associated liver disease(ALD),chronic alcohol consumption disrupts the expression and function of ncRNAs in the liver and circulation,contributing to the disease's pathogenesis and progression.Dysregulated ncRNAs influence key pathways involved in hepatocyte injury,lipid metabolism,inflam-mation,and hepatic stellate cell(HSC)activation,thereby exacerbating steatosis,inflammation,and fibrosis.Furthermore,extracellular vesicles play a pivotal role in mediating ncRNA-driven intercellular communication,amplifying liver damage and fibrosis.This review provides a comprehensive overview of the multifaceted roles of ncRNAs in ALD,with a focus on their mechanistic contributions to disease development and progression.Additionally,we discuss the potential of ncRNAs as diagnostic biomarkers and therapeutic targets,emphasizing their translational relevance in addressing the burden of ALD.

ObjectiveTo establish a geographical origin identification model for Foeniculi Fructus from Haiyuan， providing a new technical reference for the protection of Haiyuan's geo-authentic medicinal materials and its designation as a national geographical indication agricultural product. MethodsSamples of Foeniculi Fructus were collected from eight producing areas， including Minqin （Gansu）， Bozhou （Anhui）， Qingdao （Shandong）， Dezhou （Shandong）， Urumqi （Xinjiang）， Nujiang （Yunnan）， Gutuo （Inner Mongolia）， and Haiyuan （Ningxia）. Gas chromatography-ion mobility spectrometry （GC-IMS） was used to detect the volatile organic compounds （VOCs） in samples from these geographic origins. VOCs were qualitatively analyzed through dual matching with the National Institute of Standards and Technology （NIST） mass spectral database and the IMS drift time database. Using the Reporter module and Gallery Plot visualization tools within the LAV analytical platform， VOC fingerprint profiles characterizing geographic origins were constructed. A non-targeted analytical strategy was adopted， and 97 VOCs detected via GC-IMS were subjected to principal component analysis （PCA） and partial least squares discriminant analysis （PLS-DA） based on their differential distribution patterns to construct an origin identification model for Foeniculi Fructus from Haiyuan region. Key discriminative markers were screened using variable importance in projection （VIP） values greater than 1. ResultsA total of 97 VOCs were identified， including alcohols， aldehydes， ketones， esters， organic acids， terpenoids， ethers， alkenes， and benzenes. The PLS-DA model， based on VOCs data obtained by GC-IMS， effectively distinguished Foeniculi Fructus in Haiyuan region from those of other origins. During cross-validation， the model achieved a prediction parameter （Q²） of 0.976 and a goodness-of-fit parameter （R²） of 0.936， with no overfitting observed in permutation testing. Twelve key flavor markers with VIP > 1 were identified as characteristic indicators of Haiyuan origin. ConclusionA stable and highly predictive origin identification model for Foeniculi Fructus from Haiyuan was successfully established using GC-IMS technology， PLS-DA， and VIP-based marker screening. This model provides a novel technical strategy for accurately distinguishing Foeniculi Fructus in Haiyuan region from other regional varieties and offers new technical support for its protection as a geo-authentic medicinal material and a nationally designated geographical indication agricultural product in China.

ObjectiveTo establish a geographical origin identification model for Foeniculi Fructus from Haiyuan， providing a new technical reference for the protection of Haiyuan's geo-authentic medicinal materials and its designation as a national geographical indication agricultural product. MethodsSamples of Foeniculi Fructus were collected from eight producing areas， including Minqin （Gansu）， Bozhou （Anhui）， Qingdao （Shandong）， Dezhou （Shandong）， Urumqi （Xinjiang）， Nujiang （Yunnan）， Gutuo （Inner Mongolia）， and Haiyuan （Ningxia）. Gas chromatography-ion mobility spectrometry （GC-IMS） was used to detect the volatile organic compounds （VOCs） in samples from these geographic origins. VOCs were qualitatively analyzed through dual matching with the National Institute of Standards and Technology （NIST） mass spectral database and the IMS drift time database. Using the Reporter module and Gallery Plot visualization tools within the LAV analytical platform， VOC fingerprint profiles characterizing geographic origins were constructed. A non-targeted analytical strategy was adopted， and 97 VOCs detected via GC-IMS were subjected to principal component analysis （PCA） and partial least squares discriminant analysis （PLS-DA） based on their differential distribution patterns to construct an origin identification model for Foeniculi Fructus from Haiyuan region. Key discriminative markers were screened using variable importance in projection （VIP） values greater than 1. ResultsA total of 97 VOCs were identified， including alcohols， aldehydes， ketones， esters， organic acids， terpenoids， ethers， alkenes， and benzenes. The PLS-DA model， based on VOCs data obtained by GC-IMS， effectively distinguished Foeniculi Fructus in Haiyuan region from those of other origins. During cross-validation， the model achieved a prediction parameter （Q²） of 0.976 and a goodness-of-fit parameter （R²） of 0.936， with no overfitting observed in permutation testing. Twelve key flavor markers with VIP > 1 were identified as characteristic indicators of Haiyuan origin. ConclusionA stable and highly predictive origin identification model for Foeniculi Fructus from Haiyuan was successfully established using GC-IMS technology， PLS-DA， and VIP-based marker screening. This model provides a novel technical strategy for accurately distinguishing Foeniculi Fructus in Haiyuan region from other regional varieties and offers new technical support for its protection as a geo-authentic medicinal material and a nationally designated geographical indication agricultural product in China.

Objective To explore the interaction mechanisms among different dimensions of online learning competence in undergraduates,providing scientific evidence for optimizing software design and online education strategies.Methods A cross-sectional study was conducted with 347 undergraduates from Kunming Medical University via an online questionnaire in October 2024.Reliability testing and exploratory factor analysis were performed using SPSS 26.0,while structural equation modeling was constructed with Amos 24.0.Results Driving force directly and positively influenced regulatory force(β=0.778,P<0.001)and reciprocal force(β=0.302,P=0.003),but had no significant direct effect on adaptive force(β=0.131,P=0.182).Regulatory force and reciprocal force both exerted direct positive effects on adaptive force(β=0.262,P=0.022;β=0.410,P<0.001).The indirect effect of driving force on adaptive force accounted for 78.1%of the total effect(P=0.007),with regulatory force showing a significant mediating role(β=0.178,P=0.023),while reciprocal force alone did not(β=0.108,P=0.060).A chain mediating effect(driving force→regulatory force→reciprocal force→adaptive force)was confirmed(β=0.124,P=0.004).Conclusion Driving force serves as the core of online learning competence,and adaptive capacity is enhanced through regulatory and reciprocal forces.To promote knowledge internalization and adaptability,it is required to optimize feedback mechanisms and deepen interactive designs in online learning platforms.

Objectives To identify key clinical factors influencing recurrence in osteosarcoma patients,to construct and validate a Nomogram-based recurrence risk prediction model,thereby providing a quantitative tool for clinical decision-making and recurrence prevention/control.Methods Clinical data of 469 osteosarcoma patients admitted to Yunnan Cancer Hospital between 2013～2022 were retrospectively collected.Statistical analysis was performed using R software(version 4.3.2).Potential influencing factors were initially screened via univariate analysis and LASSO regression analysis.Independent predictors of osteosarcoma recurrence were then identified using multivariate logistic regression analysis.Based on the identified independent factors,a Nomogram prediction model for recurrence risk was constructed.The area under the receiver operating characteristic curve(AUC)was used to evaluate the model's discriminative ability.Results Among the entire cohort,68 patients experienced recurrence,yielding a recurrence rate of 14.50%.Multivariate analysis identified the following as independent predictors of recurrence:Primary Tumor Location:Tibial lesions(P=0.009)were associated with a significantly lower recurrence risk compared to femoral lesions(OR=0.297),while lesions in"Other Bones"(P=0.008)carried a significantly higher risk(OR=3.294).Biopsy Method:Needle biopsy(P=0.033)was associated with a significantly lower recurrence risk compared to open biopsy(OR=0.461).Lung Metastasis Status:Patients with lung metastasis(P<0.001)had a significantly higher recurrence risk than those without(OR=11.873).Lymphocyte Count:A higher lymphocyte count(P=0.001)was a protective factor,associated with a lower recurrence risk(OR=0.450).The constructed Nomogram prediction model demonstrated excellent performance:Validation results showed an AUC=0.842(95%CI:0.806～0.875),indicating outstanding discriminative ability.Conclusions This study successfully constructed and validated a Nomogram prediction model for osteosarcoma recurrence risk integrating key clinical factors.The model demonstrates superior discriminative ability and can accurately and quantitatively assess the recurrence risk for individual patients.This tool thus provides critical reference for guiding clinical treatment decisions.