Analysis of the brain transcriptome for substance-associated genes: An update on large-scale genome-wide association studies.

Substance use disorder (SUD) arises from the initiation to subsequent regular, irregular and harmful use of substances such as alcohol, tobacco/nicotine and cannabis. While thousands of genetic variants have been identified from recent large-scale genome-wide association studies (GWAS), understanding their functions in substance involvement and investigating the mechanisms by which they act in the addiction circuits remains challenging. In this study, we re-analysed the brain regional transcriptome data from the most comprehensive postmortem transcriptomic study, with a focus on up- or down-regulation of substance-associated protein-coding genes in the addiction circuit-related brain regions (AddictRegions), including six cortical and 11 subcortical regions. We found that substance-associated genes were overrepresented in AddictRegions. Interestingly, we observed a greater degree of genetic overlap between initiation and use and between use and SUD than between initiation and SUD. Moreover, substance initiation, use and SUD-associated genes tend to shift their enriched AddictRegions from the cortical for initiation and, to a lesser extent, substance use to subcortical regions for SUD (e.g., thalamus, substantia nigra and ventral tegmental area). We also uncovered a pattern of coordinated cortical up-regulation and subcortical down-regulation for the genes associated with tobacco initiation and alcohol use. Moreover, the Gene Ontology terms of glutamate receptor activity and neurotransmitter binding were most significantly overrepresented in AddictRegion-upregulated, substance-associated genes, with the strongest enrichment for those involved in common substance use behaviours. Overall, our analysis provides a resource of AddictRegion-related transcriptomes for studying substance-associated genes and generates intriguing insights into the genetic control of substance initiation, use and SUD.

A fiber-enriched diet alleviates Staphylococcus aureus-induced mastitis by activating the HDAC3-mediated antimicrobial program in macrophages via butyrate production in mice.

Mounting evidence suggests that the gut microbiota plays an important role in the pathogenesis of mastitis, an important disease affecting the health of lactating women and the development of the dairy industry. However, the effect of the regulation of the gut microbiota by dietary components on mastitis development remains unknown. In this study, we found that a fiber-enriched diet alleviated Staphylococcus aureus (S. au)-induced mastitis in mice, which was dependent on the gut microbiota as depletion of the gut microbiota by antibiotics abolished this protective effect. Likewise, fecal microbiota transplantation (FMT) from high-inulin (HI)-treated mice (HIF) to recipient mice improved S. au-induced mastitis in mice. Consumption of an HI diet and HIF increased fecal short-chain fatty acid (SCFA) levels compared with the control group. Moreover, treatment with SCFAs, especially butyrate, alleviated S. au-induced mastitis in mice. Mechanistically, consumption of an HI diet enhanced the host antimicrobial program in macrophages through inhibiting histone deacetylase 3 by the production of butyrate. Collectively, our results suggest that modulation of the gut microbiota and its metabolism by dietary components is a potential strategy for mastitis intervention and serve as a basis for other infectious diseases.

Comparative Physiological and Transcriptomic Analyses Reveal Mechanisms of Exogenous Spermidine-Induced Tolerance to Low-Iron Stress in Solanum lycopersicum L.

Iron (Fe) deficiency in plants is a major problem in agriculture. Therefore, we investigated both the physiological features and molecular mechanisms of plants' response to low-Fe (LF) stress along with the mitigation of LF with exogenous spermidine (Spd) in tomato plants. The results showed that exogenous Spd foliar application relieved the suppressing effect of LF stress on tomato plants by regulating the photosynthetic efficiency, chlorophyll metabolism, antioxidant levels, organic acid secretion, polyamine metabolism and osmoregulatory systems. Analysis of transcriptomic sequencing results revealed that the differentially expressed genes of iron-deficiency stress were mainly enriched in the pathways of phytohormone signaling, starch and sucrose metabolism and phenyl propane biosynthesis in both leaves and roots. Moreover, Spd-induced promotion of growth under LF stress was associated with upregulation in the expression of some transcription factors that are related to growth hormone response in leaves (GH3, SAUR, ARF) and ethylene-related signaling factors in roots (ERF1, ERF2). We propose that traits associated with changes in low-iron-tolerance genes can potentially be used to improve tomato production. The study provides a theoretical basis for dealing with the iron deficiency issue to develop efficient nutrient management strategies in protected tomato cultivation.

Ferritinophagy is involved in Bisphenol A-induced ferroptosis of renal tubular epithelial cells through the activation of the AMPK-mTOR-ULK1 pathway.

Bisphenol A (BPA) is a common environmental contaminant, whose exposure is associated with the progression of various kidney diseases. BPA exposure has turned out to be associated with cytotoxicity to renal tubular epithelial cells, but its underlying mechanism remains unknown. Herein, we found that BPA induced ferroptosis in kidney and renal tubular epithelial cells, as showed by increased intracellular iron accumulation, lipid peroxidation and cells death upon BPA exposure. Additionally, utilization of ferrostatin-1 and desferrioxamine, typical ferroptosis inhibitors, can fundamentally diminish cells death. Intriguingly, we discovered that autophagy inhibitor chloroquine can shield renal tubular epithelial cells from BPA-caused ferroptosis. Furthermore, we found that ferritinophagy, a phenomenon that degradation of ferritin and inducing subsequent iron overload, occurred after BPA exposure and excessive iron promoted ferroptosis through Fenton reaction. We next demonstrated that BPA activated the AMPK-mTOR-ULK1 signaling pathway. In turn, AMPK, mTOR, and ULK1 knockdown dramatically mitigated BPA-induced TCMK-1 cells death, and decreased MDA and LC3 levels, but increased FTH protein content. These results indicate that activation of the AMPK-mTOR-ULK1 signaling is involved in BPA-induced ferritinophagy. In conclusion, renal dysfunction and renal tubular epithelial damage induced by BPA are linked to ferroptosis, which depends on the activation of ferritinophagy through AMPK-mTOR-ULK1 axis.

Associations Between Smoking Status and Physical and Mental Health-Related Quality of Life Among Individuals With Mobility Impairments.

BACKGROUND: In the general population, quitting smoking is associated with improved health-related quality of life (QoL), but this association has not been examined in smokers with chronic mobility impairments (MIs). PURPOSE: We examined associations between smoking status and health-related QoL over 6 months, and whether relationships are moderated by depression and MI severity. METHODS: This is a secondary analysis of a smoking cessation induction trial among smokers with MIs (n = 241, 56% female, 36% Black) assessed at baseline, and 4 and 6 months after. Participants were grouped into "Smokers" (smoking at 4 and 6 months), "Abstainers" (quit at 4 and 6 months), "Relapsers" (relapsed at 6 months), and "Late-quitters" (quit at 6 months). Physical and mental health-related QoL was assessed with the Short-Form Health Survey. Depression was defined as scores ≥10 on the Patient Health Questionnaire, and MI severity by the use of skilled care for personal needs. Data were analyzed with linear mixed models. RESULTS: Aggregating across time, among nondepressed participants, compared with "Smokers," the "Abstainer," and "Late-quitter" groups improved their physical health scores. "Late-quitters" also improved compared with "Relapsers." Among the total sample, compared with "Smokers," "Abstainers" showed improvements in mental health scores overtime, whereas "Relapsers" improved their score at 4 months, and "Late-quitters" improved at 6 months. CONCLUSIONS: Quitting smoking is associated with improvements in physical health-related QoL regardless of the severity of MI but only among those without depression at baseline. For mental health-related QoL, associations with quitting smoking were independent of baseline depression and severity of MI.

An open-label, randomized trial of the combination of IFN-κ plus TFF2 with standard care in the treatment of patients with moderate COVID-19.

BACKGROUND: Epidemic outbreaks caused by SARS-CoV-2 are worsening around the world, and there are no target drugs to treat COVID-19. IFN-κ inhibits the replication of SARS-CoV-2; and TFF2 is a small secreted polypeptide that promotes the repair of mucosal injury and reduces the inflammatory responses. We used the synergistic effect of both proteins to treat COVID-19. METHODS: We conducted an open-label, randomized, clinical trial involving patients with moderate COVID-19. Patients were assigned in a 1:1 ratio to receive either aerosol inhalation treatment with IFN-κ and TFF2 every 24 h for six consecutive dosages in addition to standard care (experimental group) or standard care alone (control group). The primary endpoint was the time until a viral RNA negative conversion for SARS-CoV-2 in all clinical samples. The secondary clinical endpoint was the time of CT imaging improvement. Data analysis was performed per protocol. This study was registered with chictr.org.cn, ChiCTR2000030262. FINDINGS: Between March 23 and May 23 of 2020, 86 COVID-19 patients with symptoms of moderate illness were recruited, and 6 patients were excluded due to not matching the inclusion criteria (patients with pneumonia through chest radiography). Among the remaining 80 patients, 40 patients were assigned to experimental group, and the others were assigned to control group to only receive standard care. Efficacy and safety were evaluated for both groups. The time of viral RNA negative conversion in experimental group (Mean, 3·80 days, 95% CI 2·07-5·53), was significantly shorter than that in control group (7·40 days, 95% CI 4·57 to 10·23) (p = 0.031), and difference between means was 3·60 days. The percentage of patients in experimental group with reversion to negative viral RNA was significantly increased compared with control group on all sampling days (every day during the 12-day observation period) (p = 0·037). For the secondary endpoint, the experimental group had a significantly shorter time until improvement was seen by CT (Mean 6·21 days, N = 38/40, 95% CI 5·11-7·31) than that in control group (8·76 days, N = 34/40, 95% CI 7·57-9·96) (p = 0.002), and difference between means was 2·55 days. No discomfort or complications during aerosol inhalation were reported to the nurses by any experimental patients. INTERPRETATION: In conclusion, we found that aerosol inhalation of IFN-κ plus TFF2 in combination with standard care is safe and superior to standard care alone in shortening the time up to viral RNA negative conversion in all clinical samples. In addition, the patients in experimental group had a significantly shortened CT imaging improvement time than those in control group. This study suggested that this combination treatment is able to facilitate clinical improvement (negative for virus, improvement by CT, reduced hospitalization stay) and thereby result in an early release from the hospital. These data support the need for exploration with a large-scale trial of IFN-κ plus TFF2 to treat COVID-19. FUNDING: Funding was provided by the National Natural Science Foundation of China, National Major Project for Control and Prevention of Infectious Disease in China, Shanghai Science and Technology Commission, Shanghai Municipal Health Commission.

Opioid Prescribing Patterns Among Oral and Maxillofacial Surgeons: A Regional Survey-Based Study.

PURPOSE: We sought to obtain baseline statistics regarding the amount of opioid tablets prescribed by oral and maxillofacial surgeons (OMSs) in the New England area after office-based procedures and to identify factors that might be predictors of their prescription patterns. MATERIALS AND METHODS: An anonymous online survey was e-mailed to practicing OMSs in the New England area. The survey explored the quantity of opioid medications prescribed for various procedures, how opioid precautions were given, practitioners' attitude toward opioid dependency, and whether certain surgeon- or patient-related factors influenced prescription behavior. Statistical analyses were used to categorize the OMSs according to their prescription patterns and to identify the most common factors affecting their decision to prescribe opioids. RESULTS: Of 315 OMSs, 151 (43%) responded to the survey. Our analyses were of complete data obtained from 118 respondents. For procedures, such as extraction of 7 or more teeth, the placement of 4 or more implants, office-based sinus surgery, cortical block grafts, and removal of third molar teeth, respondents indicated they typically prescribed 8 to 12 opioid tablets. For all other procedures, they typically never or rarely prescribed opioid tablets. The respondents were grouped into low-, medium-, and high-quantity opioid prescribers. Regardless of their grouping status, the respondents showed general agreement regarding their roles in reducing opioid prescription-related issues. No group differences were found in terms of the demographic variables. Relative to the factors predicting increased prescribing habits, the results suggested that OMSs working either exclusively or primarily in academic settings tended to prescribe fewer opioid tablets than those working primarily in the private setting (ß = -2.73; P < .001). Additionally, 109 respondents (92.4%) reported that OMSs could play a role in decreasing opioid dependency. CONCLUSIONS: Most practicing OMSs in the New England area prescribed opioids after office-based surgery and are cognizant of the risks of opioid medications.

Modulation of the Pol II CTD Phosphorylation Code by Rac1 and Cdc42 Small GTPases in Cultured Human Cancer Cells and Its Implication for Developing a Synthetic-Lethal Cancer Therapy.

Rho GTPases, including Rho, Cdc42, Rac and ROP subfamilies, are key signaling molecules in RNA polymerase II (Pol II) transcriptional control. Our prior work has shown that plant ROP and yeast Cdc42 GTPases similarly modulate Ser2 and Ser5 phosphorylation status of the C-terminal domain (CTD) of the Pol II largest subunit by regulating CTD phosphatase degradation. Here, we present genetic and pharmacological evidence showing that Cdc42 and Rac1 GTPase signaling modulates a similar CTD Ser2 and Ser5 phosphorylation code in cultured human cancer cells. While siRNA knockdown of Cdc42 and Rac1, respectively, in HeLa cells increased the level of CTD Ser phosphatases RPAP2 and FCP1, they both decreased the level of CTD kinases CDK7 and CDK13. In addition, the protein degradation inhibitor MG132 reversed the effect of THZ1, a CDK7 inhibitor which could decrease the cell number and amount of CDK7 and CDK13, accompanied by a reduction in the level of CTD Ser2 and Ser5 phosphorylation and DOCK4 and DOCK9 (the activators for Rac1 and Cdc42, respectively). Conversely, treatments of Torin1 or serum deprivation, both of which promote protein degradation, could enhance the effect of THZ1, indicating the involvement of protein degradation in controlling CDK7 and CDK13. Our results support an evolutionarily conserved signaling shortcut model linking Rho GTPases to Pol II transcription across three kingdoms, Fungi, Plantae and Animalia, and could lead to the development of a potential synthetic-lethal strategy in controlling cancer cell proliferation or death.

Opioid prescribing patterns of oral and maxillofacial surgery residents.

OBJECTIVE: The aim of this study was to report on the opioid prescribing patterns of oral and maxillofacial surgery residents for routine office-based procedures. STUDY DESIGN: A survey was sent to current resident members of the American Association of Oral and Maxillofacial Surgeons (AAOMS). The survey asked questions related to the quantity and duration of opioids prescribed and if such factors as type of procedure, education, and prior opioid abuse by patients influenced prescription behavior. RESULTS: There was a positive relationship between the prescribing culture of residency and prescribing pattern (P value < .001). Concerns about factors that promote addiction were negatively associated with prescribing patterns (P value < .02). Prescribing culture and concerns about factors promoting addiction together explained 10% of the total variation. Residents receiving instruction on the risks of narcotic were less likely to prescribe opioids. CONCLUSIONS: Creating a culture in residency programs aimed at reducing factors that promote addiction may be the best method to reduce overprescription of opioids during training.

Contrast-enhanced ultrasonography of nasopharyngeal carcinoma: enhancement patterns and quantitative parameters.

OBJECTIVE: The aim of this study was to investigate the characteristics of nasopharyngeal carcinoma (NPC) using contrast-enhanced ultrasonography (CEUS), including the enhancement patterns and the quantitative parameters. METHODS: Having been scanned using conventional ultrasonography (US) and CEUS, every case was confirmed to be NPC under endoscopic biopsy, and no case received any anti-tumor treatment before CEUS examinations. Tumor/node/metastasis stages were determined in accordance with 2002 AJCC 6th edition. Contrast enhancement patterns and quantitative parameters were observed. RESULTS: CEUS imaging of NPC showed that the tumor signal intensity enhanced early, rapidly, and remarkably, and decreased slowly later. The patterns of enhancement included spot/linear enhancement, peripheral enhancement, and mass enhancement, and two types of time intensity curves of NPC included type I and type II. There was a significant difference between peak intensity (PI) and T stage (P<0.05), whereas time-to-peak (TP) and slope did not show significant differences with T stage (P>0.05). CONCLUSION: CEUS is feasible to be applied to the nasopharynx region. The use of CEUS makes it possible to observe vascular permeability of NPC. Our results suggest that the quantitative parameter PI of nasopharyngeal carcinoma is significantly different from T stages. Thus, PI may serve as a potential noninvasive radiological prognostic indicator for NPC.

Using Calibration to Reduce Measurement Error in Prevalence Estimates Based on Electronic Health Records.

INTRODUCTION: Increasing adoption of electronic health record (EHR) systems by health care providers presents an opportunity for EHR-based population health surveillance. EHR data, however, may be subject to measurement error because of factors such as data entry errors and lack of documentation by physicians. We investigated the use of a calibration model to reduce bias of prevalence estimates from the New York City (NYC) Macroscope, an EHR-based surveillance system. METHODS: We calibrated 6 health indicators to the 2013-2014 NYC Health and Nutrition Examination Survey (NYC HANES) data: hypertension, diabetes, smoking, obesity, influenza vaccination, and depression. We classified indicators into having low measurement error or high measurement error on the basis of whether the proportion of misclassification (ie, false-negative or false-positive cases) was greater than 15% in 190 reviewed charts. We compared bias (ie, absolute difference between NYC Macroscope estimates and NYC HANES estimates) before and after calibration. RESULTS: The health indicators with low measurement error had the same bias after calibration as before calibration (diabetes, 2.5 percentage points; smoking, 2.5 percentage points; obesity, 3.5 percentage points; hypertension, 1.1 percentage points). For indicators with high measurement error, bias decreased from 10.8 to 2.5 percentage points for depression, and from 26.7 to 8.4 percentage points for influenza vaccination. CONCLUSION: The calibration model has the potential to reduce bias of prevalence estimates from EHR-based surveillance systems for indicators with high measurement errors. Further research is warranted to assess the utility of the current calibration model for other EHR data and additional indicators.

Aluminum alkoxide, amide and halide complexes supported by a bulky dipyrromethene ligand: synthesis, characterization, and preliminary ε-caprolactone polymerization activity.

Aluminum halide, alkoxide and amide complexes 2-6 of the form (N,N)AlX2-nYn (n = 0, 1 and (N,N) = 1,9-dimesityl-5-phenyldipyrromethene (1)) were synthesized and characterized by NMR spectroscopy and X-ray crystallography. The in situ generated lithium salt of dipyrromethene 1 was reacted with AlX3 to afford aluminum halide complexes (N,N)AlX2 (X = Cl (2), I (3)) which were isolated as dichroic crystals. Salt metathesis reactions were employed to produce alkoxide complexes (N,N)Al(Cl)(O(t)Bu) (4) and (N,N)Al(O(t)Bu)2 (5) from compound 2. The dimethylamide complex (N,N)Al(NMe2)2 (6) was prepared by reaction of dipyrromethene 1 with [Al(NMe2)3]2. Crystallographic data revealed that the dipyrromethene is non-planar when bulky coligands are present as in compounds 3-6, while in the dichloride complex 2 the dipyrromethene is planar. Halide complexes 2 and 3 reacted with adventitious moisture in toluene to afford crystalline acid-base adducts (N,N)H·HX, (X = Cl (7), I (8)), which adopted structures reminiscent of anion receptors. Alkoxide and dimethylamide complexes 5 and 6 were also applied as precatalysts for the ring-opening polymerization of ε-caprolactone and preliminary results are reported.

Annual Research Review: Discovery science strategies in studies of the pathophysiology of child and adolescent psychiatric disorders--promises and limitations.

BACKGROUND: Psychiatric science remains descriptive, with a categorical nosology intended to enhance interobserver reliability. Increased awareness of the mismatch between categorical classifications and the complexity of biological systems drives the search for novel frameworks including discovery science in Big Data. In this review, we provide an overview of incipient approaches, primarily focused on classically categorical diagnoses such as schizophrenia (SZ), autism spectrum disorder (ASD), and attention-deficit/hyperactivity disorder (ADHD), but also reference convincing, if focal, advances in cancer biology, to describe the challenges of Big Data and discovery science, and outline approaches being formulated to overcome existing obstacles. FINDINGS: A paradigm shift from categorical diagnoses to a domain/structure-based nosology and from linear causal chains to complex causal network models of brain-behavior relationship is ongoing. This (r)evolution involves appreciating the complexity, dimensionality, and heterogeneity of neuropsychiatric data collected from multiple sources ('broad' data) along with data obtained at multiple levels of analysis, ranging from genes to molecules, cells, circuits, and behaviors ('deep' data). Both of these types of Big Data landscapes require the use and development of robust and powerful informatics and statistical approaches. Thus, we describe Big Data analysis pipelines and the promise and potential limitations in using Big Data approaches to study psychiatric disorders. CONCLUSIONS: We highlight key resources available for psychopathological studies and call for the application and development of Big Data approaches to dissect the causes and mechanisms of neuropsychiatric disorders and identify corresponding biomarkers for early diagnosis.

Oridonin suppress cell migration via regulation of nonmuscle myosin IIA.

Oridonin, which is isolated from Chinese herb Rabdosia rubescens (Hemsl.) Hara, has been implicated in regulation of tumor cell migration and invasion. In this study, treatment with oridonin enhanced the phosphorylation of myosin regulatory light chain (T18/S19) that regulates the ATPase activity of myosin IIA. Meanwhile, stress fibers were significantly more prominent after oridonin incubation, which impaired cell migration in transwell migration assays. All of these effects may be caused by the decreased interaction between myosin IIA and myosin phosphatase complex, but not kinases. Our data provide clear evidence of this novel pharmacological function for oridonin in treating cancer cell migration.

Aberrant gene expression in the Arabidopsis SULTR1;2 mutants suggests a possible regulatory role for this sulfate transporter in response to sulfur nutrient status.

Sulfur is required for the biosynthesis of cysteine, methionine and numerous other metabolites, and thus is critical for cellular metabolism and various growth and developmental processes. Plants are able to sense their physiological state with respect to sulfur availability, but the sensor remains to be identified. Here we report the isolation and characterization of two novel allelic mutants of Arabidopsis thaliana, sel1-15 and sel1-16, which show increased expression of a sulfur deficiency-activated gene ß-glucosidase 28 (BGLU28). The mutants, which represent two different missense alleles of SULTR1;2, which encodes a high-affinity sulfate transporter, are defective in sulfate transport and as a result have a lower cellular sulfate level. However, when treated with a very high dose of sulfate, sel1-15 and sel1-16 accumulated similar amounts of internal sulfate and its metabolite glutathione (GSH) to wild-type, but showed higher expression of BGLU28 and other sulfur deficiency-activated genes than wild-type. Reduced sensitivity to inhibition of gene expression was also observed in the sel1 mutants when fed with the sulfate metabolites Cys and GSH. In addition, a SULTR1;2 knockout allele also exhibits reduced inhibition in response to sulfate, Cys and GSH, consistent with the phenotype of sel1-15 and sel1-16. Taken together, the genetic evidence suggests that, in addition to its known function as a high-affinity sulfate transporter, SULTR1;2 may have a regulatory role in response to sulfur nutrient status. The possibility that SULTR1;2 may function as a sensor of sulfur status or a component of a sulfur sensory mechanism is discussed.

Ambulatory assessments of psychological and peripheral stress-markers predict birth outcomes in teen pregnancy.

OBJECTIVE: Pregnant adolescents have high rates of poor birth outcomes, but the causes are unclear. We present a prospective, longitudinal study of pregnant adolescents assessing associations between maternal psychobiological stress indices and offspring gestational age at birth and birthweight. METHOD: Healthy nulliparous pregnant adolescents were recruited (n=205) and followed during pregnancy. Ambulatory assessments over 24h of perceived psychological stress (collected every 30 min) and salivary cortisol (6 samples) and a summary questionnaire, the Perceived Stress Scale, were collected at three time points (13-16, 24-27, and 34-37 gestational weeks). Corticotropin-releasing hormone, C-reactive protein, and interleukin 6 were assayed from blood taken at the latter 2 sessions. A final sample of 119 participants was selected for analyses. RESULTS: The ambulatory assessment of perceived psychological stress was positively correlated with the Perceived Stress Scale (r=.20, p=.03) but neither was associated with any of the biological assays (all ps>.20). Based on backward selection regression models that included all stress variables and relevant covariates, the ambulatory assessments of perceived psychological stress and cortisol - though not the Perceived Stress Scale - were negatively associated with gestational age at birth (F(4, 107)=3.38, p=.01) while cortisol was negatively related to birthweight (F(5, 107)=14.83, p<.0001). CONCLUSIONS: Targeted interventions to reduce psychological and biological indicators of heightened stress during pregnancy may have positive public health benefits for the offspring given the associations of shortened gestation and lower birthweight with risk for poor mental and physical health outcomes.

[Effect of epimedii folium processed with different refining temperatures and amounts of sheep's oil on kidney-yang deficiency rats].

OBJECTIVE: To investigate the effects of refining temperatures and amounts of sheep's oil used in processing Epimedii Folium on Kedney-yang deficiency rats. METHOD: The corticosterone was subcutaneous injected to establish the kidney yang deficiency rat model. With the temperatures and amounts of sheep's oil were 250 degrees C/30%, 120 degrees C/30% and 120 degrees C/20% respectively, the crude drug and three kinds of pressed Epimedii Folium were extracted by water and used as examined samples while total flavonoid of Epimedii Folium was used as positive control. After examined samples and control samples were intragastirc administrated, the pharmacologic action was analyzed. RESULT: As compared to crude drug, all of the aqueous extracts of processed Epimedii Folium have stronger effect of warming kidney and enhancing yang, especially the Epimedii Folium processed by sheep's oil with refining temperatures 120 degrees C and amounts of sheep's oil 30%. Its mechanism might be related to improving the insufficiency of hypothalamic-pituitary-adrenal-thymus (HPAT) axis suppression. CONCLUSION: The refining temperature of sheep's oil can affect the quality of excipients and processed drugs. The results may be useful in explaining the mechanism of Epimedii Folium processing and establishment of pharmaceutical standard of sheep's oil used as processing excipients.