Saiga antelope horn suppresses febrile seizures in rats by regulating neurotransmitters and the arachidonic acid pathway.

BACKGROUND: Saiga antelope horn (SAH) is a traditional Chinese medicine for treating febrile seizure (FS) with precise efficacy, but its mechanism of action and functional substances are still unclear. Given the need for further research on SAH, our group conducted studies to elucidate its mechanisms and active substances. METHODS: An FS rat pup model was constructed through intraperitoneal injection of LPS and hyperthermia induction. Behavioural indicators of seizures, hippocampal histopathological alterations, serum levels of inflammatory cytokines and hippocampal levels of neurotransmitters were observed and measured to investigate the effects of SAH on FS model rats. Hippocampal metabolomics and network pharmacology analyses were conducted to reveal the differential metabolites, key peptides and pathways involved in the suppression of FS by SAH. RESULTS: SAH suppressed FS, decreased the inflammatory response and regulated the Glu-GABA balance. Metabolomic analysis revealed 13 biomarkers of FS, of which SAH improved the levels of 8 differential metabolites. Combined with network pharmacology, a "biomarker-core target-key peptide" network was constructed. The peptides of SAH, such as YGQL and LTGGF, could exert therapeutic effects via the arachidonic acid pathway. Molecular docking and ELISA results indicated that functional peptides of SAH could bind to PTGS2 target, inhibiting the generation of AA and its metabolites in hippocampal samples. CONCLUSION: In summary, the functional peptides contained in SAH are the main material basis for the treatment of FS, potentially acting through neurotransmitter regulation and the arachidonic acid pathway.

Mechanism and functional substances of Saiga antelope horn in treating hypertension with liver-yang hyperactivity syndrome explored using network pharmacology and metabolomics.

ETHNOPHARMACOLOGICAL RELEVANCE: Saiga antelope horn (SAH) is a traditional Chinese medicine for treating hypertension with liver-yang hyperactivity syndrome (Gan-Yang-Shang-Kang, GYSK), that has a long history of clinical application and precise efficacy, but its mechanism and functional substances are still unknown. Based on the demand for alternative research on the rare and endangered SAH, the group designed and carried out the following studies. AIM OF THE STUDY: The purpose of this research was to demonstrate the functional substances and mechanisms of SAH in the treatment of GYSK hypertension. MATERIALS AND METHODS: The GYSK-SHR model was constructed by administering a decoction of aconite to spontaneously hypertensive rats (SHRs). Blood pressure (BP), behavioural tests related to GYSK, and pathological changes in the kidneys, heart and aorta were measured to investigate the effects of SAH on GYSK-SHRs. Proteomic analysis was used to identify the keratins and peptides of SAH. Moreover, network pharmacology and plasma metabolomics studies were carried out to reveal the mechanisms by which functional peptides in SAH regulate GYSK-hypertension. RESULTS: SAH has a significant antihypertensive effect on GYSK hypertensive animals. It has also been proven to be effective in protecting the function and structural integrity of the kidneys, heart and aorta. Moreover, SAH improved the abnormalities of 31 plasma biomarkers in rats. By constructing a "biomarker-target-peptide" network, 10 functional peptides and two key targets were screened for antihypertensive effects of SAH. The results indicated that SAH may exert a therapeutic effect by re-establishing the imbalance of renin-angiotensin (RAS) system. CONCLUSIONS: Functional peptides from keratin contained in SAH are the main material basis for the treatment of GYSK-hypertension and exhibited the protective effect on the GYSK-SHR model through the RAS system.

Carp scales derived double cross-linking hydrogels achieve collagen peptides sustained-released for bone regeneration.

Collagen peptide exhibits a great activity in osteogenic differentiation and wound healing. However, uncontrolled collagen peptide release in bone defects leads to unsatisfactory bone regeneration. In this work, we prepared collagen peptide loaded calcium alginate hydrogel (SA-CP/Ca) derived from Asia carp scales by mixing sodium alginate solution, collagen peptides, calcium carbonate, covalent cross-linking agents N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDC), and N-hydroxysuccinimide (NHS) in one pot. Physically and chemically double cross-linking realized higher crosslink density, smaller porosity and pore size, and higher energy storage modulus and loss modulus, achieving sustained release of collagen peptides. The release profile is fitted to Keppas-Sahlin model, to find SA-CP/Ca hydrogels are more inclined to release collagen peptides through expansion and degradation. The compatibility and osteogenic ability of SA-CP/Ca are demonstrated in vitro and in vivo.

Protective effects of Lycium barbarum L. berry extracts against oxidative stress-induced damage of the retina of aging mouse and ARPE-19 cells.

In this study, we investigated the preventive effect of Lycium barbarum L. berry extract on age-related macular degeneration (AMD) and the main components responsible for its antioxidant activity. An AMD mouse model was developed by feeding 18-month-old mice with a 1% hydroquinone diet. Meanwhile, the model mice were treated with water extract (LBW) and alcohol extract (LBE) of L. barbarum berries respectively for 3 months. It was found that the retinal structural abnormalities were improved and the oxidation stress and inflammatory imbalance were both attenuated in model mice treated with the extracts of L. barbarum berries. According to the metabolomics analysis of the serum of model mice, LBW regulated the metabolism of unsaturated fatty acids and sphingolipids, while LBE extracts tended to regulate taurine metabolism. On sodium iodate induced oxidative injury of ARPE-19 cells, water extracts of L. barbarum berries eluted with 95% ethanol (LBW-95E) on AB-8 macroporous resin significantly improved the cell viability and attenuated oxidative stress by increasing the superoxide dismutase (SOD) activity and glutathione (GSH) content, decreasing the reactive oxygen species (ROS) content, promoting the entry of nuclear factor erythroid-derived 2-like 2 (Nrf2) into the nucleus and up-regulating the heme oxygenase-1 (HO-1) expression. Scopoletin, N-trans-feruloyltyramine and perlolyrine were identified as the main components of LBW-95E. These results demonstrated that L. barbarum berry extracts protected the retina of aging AMD model mice from degeneration and LBW-95E was the vital antioxidant activity fraction of LBW. These findings suggest that L. barbarum berry extracts might be an excellent natural source for the development of retinal protection-related drugs or dietary supplements.

EGFR activation-induced decreases in claudin1 promote MUC5AC expression and exacerbate asthma in mice.

Claudin1 plays a critical role in maintaining the epithelial barrier, and mucus hypersecretion induced by epidermal growth factor receptor (EGFR) activation is a pivotal pathological feature of asthma. The relationship between claudin1 expression and mucus hypersecretion and EGFR activation is still poorly understood. In this report, we showed that claudin1 expression correlated with asthma stage, in both patients with asthma and in the house dust mite (HDM)-induced mouse asthma model. Claudin1 knockdown induced MUC5AC overexpression both in 16HBE cells and in mouse airways. In addition, claudin1 expression negatively correlated with asthma severity as demonstrated by significantly higher MUC5AC expression, more severe airway inflammation, and increased airway hyperreactivity in mouse lungs with claudin1 knockdown following HDM challenge. EGFR activation reduced claudin1 expression and increased MUC5AC expression, both in vitro and in vivo. Erlotinib alleviated murine allergic airway inflammation, restored claudin1 expression and decreased MUC5AC expression. These results suggest that EGFR activation-induced decreases in claudin1 promote goblet-cell metaplasia, and restoring claudin1 to maintain barrier integrity by EGFR antagonism may provide a novel therapeutic strategy for asthma.

Frontline Science: Two flavonoid compounds attenuate allergic asthma by regulating epithelial barrier via G protein-coupled estrogen receptor: Probing a possible target for allergic inflammation.

Allergic asthma is a common chronic lung inflammatory disease and seriously influences public health. We aim to investigate the effects of formononetin (FMN) and calycosin (CAL), 2 flavonoids in Radix Astragali, on allergic asthma and elucidate possible therapeutic targets. A house dust mite (HDM)-induced allergic asthma mouse model and TNF-α and Poly(I:C) co-stimulated human bronchial epithelial cell line (16HBE) were performed respectively in vivo and in vitro. The role of G protein-coupled estrogen receptor (GPER) was explored by its agonist, antagonist, or GPER small interfering RNA (siGPER). E-cadherin, occludin, and GPER were detected by western blotting, immunohistochemistry, or immunofluorescence. The epithelial barrier integrity was assessed by trans-epithelial electric resistance (TEER). Cytokines were examined by enzyme-linked immunosorbent assay (ELISA). The results showed that flavonoids attenuated pulmonary inflammation and hyperresponsiveness in asthmatic mice. These flavonoids significantly inhibited thymic stromal lymphopoietin (TSLP), increased occludin and restored E-cadherin in vivo and in vitro. The effects of flavonoids on occludin and TSLP were not interfered by ICI182780 (estrogen receptor antagonist), while blocked by G15 (GPER antagonist). Furthermore, compared with PPT (ERα agonist) and DPN (ERß agonist), G1 (GPER agonist) significantly inhibited TSLP, up-regulated occludin, and restored E-cadherin. siGPER and TEER assays suggested that GPER was pivotal for the flavonoids on the epithelial barrier integrity. Finally, G1 attenuated allergic lung inflammation, which could be abolished by G15. Our data demonstrated that 2 flavonoids in Radix Astragali could alleviate allergic asthma by protecting epithelial integrity via regulating GPER, and activating GPER might be a possible therapeutic strategy against allergic inflammation.

Chinese medicine Yu-Ping-Feng-San attenuates allergic inflammation by regulating epithelial derived pro-allergic cytokines.

This study aimed to investigate the mechanisms of Yu-Ping-Feng-San (YPFS) on attenuating allergic inflammation in the initial stage of atopic dermatitis (AD). AD mouse model was established with fluorescein isothiocyanate (FITC) sensitization and elicitation. Epithelial barrier structure was observed with transmission electron microscope. The populations of dendritic cells (DCs) and group 2 innate lymphoid cells (ILC2s) were detected by flow cytometry. Human immortalized keratinocyte (HaCaT) cells were stimulated with Poly(I:C)/TNF-α in vitro to assessthymic stromal lymphopoietin (TSLP), interleukin (IL)-33 and nuclear factor-κB (NF-κB) levels or expressions by immunofluorescence, enzyme linked immunosorbent assay (ELISA) and western blot. In the initial stage of AD, ear swelling and infiltration of inflammatory cells in ear tissues were markedly attenuated with YPFS treatments. The damaged structures of ear epithelium and the increased levels of Th2-cytokines induced by FITC were significantly rescued in YPFS-treated mice. The production of pro-allergic cytokines, TSLP and IL-33, as well as the cell populations of their target cells DCs and ILC2s were decreased in AD model, respectively. Likewise, the levels of TSLP and IL-33 in Poly(I:C)/TNF-α-stimulated HaCaT cells showed the same results. Lower levels of p-NF-κB were detected with YPFS treatment, and the expressions of TSLP and IL-33 could be further decreased with inhibiting of NF-κB. Therefore, YPFS attenuates allergic inflammation in the initial stage of AD probably through regulating NF-κB-TSLP/IL-33 pathway, which may provide a novel effective target for the prevention and treatment of allergic diseases.

Formononetin attenuated allergic diseases through inhibition of epithelial-derived cytokines by regulating E-cadherin.

Radix Astragali, has long been used to alleviate allergic diseases (ADs). Formononetin is one of the major active components in Radix Astragali, but its mechanism on ADs is not definitively known. The fluorescein isothiocyanate isomer-induced atopic contact dermatitis mouse model and poly I:C or lipopolysaccharide-treated HaCaT cells were used to examine thymic stromal lymphopoietin (TSLP)/interleukin (IL)-33 production and expression of E-cadherin. After administration of formononetin, TSLP/IL-33 levels decreased both in vitro and in vivo, while E-cadherin was increased in vivo and restored in vitro. Furthermore, small interference RNA silencing of E-cadherin resulted in elevated levels of TSLP, whereas the inhibitory effect of formononetin on TSLP was no longer observed. In addition, TSLP resulted in no detectable changes in delocalization or protein expression of E-cadherin in HaCaT cells. These results indicated that formononetin showed a protective effect in ADs, which was correlated with decreasing TSLP/IL-33 production via regulation of E-cadherin.

[Effect and mechanism of aerial parts of Salvia miltiorrhiza effective constituents on glycolipid metabolism of high sugar-induced Drosophila melanogaster metabolic disorder model].

To evaluate the effect and mechanism of aerial parts of Salvia miltiorrhiza(SM) on high sugar-induced Drosophila melanogaster metabolic disorder model. The levels of glucose, triglyceride and protein in SM were detected; nymphosis time was recorded, and the reliability of metabolic disorder model as well as the mechanism of aerial parts of SM were evaluated based on metabonomics. The results showed that the levels of glucose and triglyceride in model group were significantly higher than those in normal control group(P<0.05). As compared with the model group, the glucose level was significantly decreased in gliclazide(GLZ) group, SM medium(SM-M) and high(SM-H) dose groups(P<0.05, P<0.01); the triglyceride level was significantly decreased in GLZ group and SM-H group(P<0.05, P<0.01). By principal component analysis(PCA) and partial least squares discriminant analysis(PLS-DA), the metabolic level of model ones was recovered to a certain degree after intervention by aerial parts of SM. Seventeen marker compounds and four major metabolic pathways were obtained by screening differential metabolites, comparing literature and retrieving the database. The aerial parts of SM may regulate glycolipid metabolism through the impact on histidine metabolism, glycerophospholipid metabolism, pentose and glucuronate interconversions, cysteine and methionine metabolism and glycerolipid metabolism. Extract from aerial parts of SM can regulate the glycolipid metabolism of D. melanogaster metabolic disorder model and make it return to normal condition. This paper provides reference for the value discovery and resource utilization of the aerial parts of S. miltiorrhiza.

Gancao-Gansui combination impacts gut microbiota diversity and related metabolic functions.

ETHNOPHARMACOLOGICAL RELEVANCE: The theory of "eighteen incompatible medicaments" (EIM) in traditional Chinese medicine (TCM) is the most representative case of herbal-herbal interactions. Gancao and Gansui are one of the incompatible herbal pairs in EIM. Gancao, also known as "licorice", is the most frequently used Chinese herb or food additive. Gansui, the root of Euphorbia kansui T.P. Wang, is another famous Chinese herb usually used to treat edema, ascites and asthma but could induce gastrointestinal (GI) tract irritation. Although Gancao and Gansui are incompatible herbal pairs, they are still used in combination in the famous "Gansui-Banxia" decoction. AIM OF THE STUDY: This study was conducted to investigate if Gancao-Gansui combination could exacerbate Gansui induced GI tract injury. Moreover, the impact of Gancao-Gansui combination to gut microbiota and related metabolism pathways were evaluated. MATERIALS AND METHODS: Normal mice were divided into different groups and treated with Gancao extracts, Gansui extracts, and Gancao-Gansui combination extracts for 7 days. Serum biomarkers (diamine oxidase activity, lipopolysaccharide, motilin, IL-1ß, IL-6, TNF-α) were determined to reflect GI tract damage. Gut microbiota diversity was studied by 16S rDNA sequencing and metagenomes analysis were also conducted to reflect functional genes expression alteration. Fecal hydrogen sulfide concentrations were measured by spectrophotometry to confirm the alteration of Desulfovibrio genus. Fecal lipid metabolomics study was conducted by GC-MS analysis to confirm the change of metagenomes and Mycoplasma abundance. RESULTS: Gancao-Gansui combination did not exacerbate GI tract tissue or functional damage but caused gut microbiota dysbiosis and increased some rare genus's abundance including Desulfovibrio and Mycoplasma. Desulfovibrio genus proliferation was confirmed by the disturbance of fecal hydrogen sulfide homeostasis. Gancao-Gansui combination also dys-regulated the metabolic genes in metagenomes. Mycoplasma genus proliferation and the metagenomes changes were both confirmed by metabolic profile analysis of fecal lipids, especially cholesterol. CONCLUSIONS: Gancao-Gansui combination can impact the gut microbiota diversity and related metabolic functions. Further studies should be carried out when the combination of Gancao-Gansui is used in herbal formulations as this may alter the diversity of the microbiota.

How impaired efficacy happened between Gancao and Yuanhua: Compounds, targets and pathways.

As recorded in Traditional Chinese Medicine (TCM) theory, Gancao (Glycyrrhizae Radix et Rhizoma) could weaken the pharmacological effect or increase the toxicity of Yuanhua (Genkwa Flos). However, the theory has been suspected due to lack of evidence. Here, we investigate whether Gancao could weaken Yuanhua's diuretic effect, if so, which chemicals and which targets may be involved. Results showed that Yuanhua exerted diuretic effect through down-regulating renal AQP 2, without electrolyte disturbances such as K+ loss which has been observed as side-effect of most diuretics. Gancao had no diuretic effect, but could impair Yuanhua's diuretic effect through up-regulating renal AQP 2. Glycyrrhetinic acid (GRA) in Gancao could up-regulate AQP 2 and counteract the AQP 2 regulation effect of Yuanhuacine (YHC) and Ginkwanin (GKW) in Yuanhua. Network pharmacology method suggested that YHC, GKW and GRA could bind to MEK1/FGFR1 protein and influence ERK-MAPK pathway, which was verified by Western blotting. This study supports TCM theory and reminds that more attention should be paid to the safety and efficacy problems induced by improper combination between herbs. Moreover, we suggested that promising diuretics with less side effects can be developed from Chinese Medicines such as Yuanhua.

The Adverse Effects of Triptolide on the Reproductive System of Caenorhabditis elegans: Oogenesis Impairment and Decreased Oocyte Quality.

Previous studies have revealed that Triptolide damages female reproductive capacity, but the mechanism is unclear. In this study, we used Caenorhabditis elegans to investigate the effects of Triptolide on the germline and explore its possible mechanisms. Our data show that exposure for 4 h to 50 and 100 mg/L Triptolide reduced C. elegans fertility, led to depletion and inactivation of spermatids with the changes in the expression levels of related genes, and increased the number of unfertilized oocytes through damaging chromosomes and DNA damage repair mechanisms. After 24 and 48 h of the 4 h exposure to 50 and 100 mg/L Triptolide, we observed shrink in distal tip cells, an increase in the number of apoptotic cells, a decrease in the number of mitotic germ cells and oocytes in diakinesis stage, and chromatin aggregates in -1 oocytes. Moreover, expression patterns of the genes associated with mitotic germ cell proliferation, apoptosis, and oocyte quality were altered after Triptolide exposure. Therefore, Triptolide may damage fertility of nematodes by hampering the development of oocytes at different developmental stages. Alterations in the expression patterns of genes involved in oocyte development may explain the corresponding changes in oocyte development in nematodes exposed to Triptolide.

Hierarchical identification of bioactive components in a medicinal herb by preparative high-performance liquid chromatography and selective knock-out strategy.

A novel and generally applicable approach was established to hierarchically identify the bioactive components of a medicinal herb by preparative high-performance liquid chromatography (prep-HPLC) and a selective knock-out strategy. In this study, the targeted components of an herbal medicine were separated and knocked out using prep-HPLC. Subsequently, the contributions of the different target components to the overall effect of the medicinal herb were comparatively evaluated and differentiated by a heat map and a 3D score plot. This approach was successfully applied to investigate the bioactive constituents of safflower. The contributions of 11 components to the overall effect of safflower were as follows: anhydrosafflor yellow B (10)>6-hydroxykaempferol 3,6-di-O-ß-d-glucoside (8)>hydroxysafflor yellow A (3)>kaempferol 3-O-ß-rutinoside (11)>6-hydroxykaempferol 3-O-ß-rutinoside (9)>6-hydroxykaempferol 3,6-di-O-ß-d-glucoside-7-O-ß-d-glucuronide (4)>6-hydroxyapigenin 6-O-ß-d-glucoside-7-O-ß-d-glucuronide (6)>cytidine (1)>6-hydroxykaempferol 3-O-ß-rutinoside-6-O-ß-d-glucoside (7)>6-hydroxykaempferol 3,6,7-tri-O-ß-d-glucoside (5)>adenosine (2). These results demonstrate that quinochalcone C-glycosides (3 and 10) and some flavonoid glycosides containing C7-OH (such as 8, 9 and 11) made a greater contribution to the overall effect of safflower than the other components that were knocked out. The results provided an important reference for improving quality control and further development of safflower products. And this approach should also be useful for investigating the bioactive constituents of other medicinal herbs.

The mixture of Salvia miltiorrhiza-Carthamus tinctorius (Danhong injection) alleviates low-dose aspirin induced gastric mucosal damage in rats.

BACKGROUND: Danhong injection (DHI) is quite often used in combination with low-dose aspirin (ASA, 75-325mg daily) in clinic, particularly for the treatment of cardiovascular diseases. Exploring their interaction profile is of great clinical importance. PURPOSE: The current study aims to explore the interaction between DHI and low-dose ASA in rats. METHODS: Sixty four rats were randomly divided into eight groups. Stomach and other four vital organs were collected for histological evaluation. Organs which exhibited histological changes were selected for a further study to evaluate the damage score and mode of action. We tested the protective effect of DHI on gastric mucosal damage in different regimes of administration. COX activity, gastric mucus secretion, pepsin activity, antioxidant activity and ROS level were assayed to reflect the protective effect of DHI on gastric mucosal damage induced by ASA. RESULTS: Stomach was the target organ of interaction when DHI and ASA were used in combination. DHI alleviated gastric mucosal damage by 55.8% when DHI was injected before ASA (Group E) and by 53.5% when DHI was injected 2h after ASA administration (Group F). Additionally, if DHI treatment was appended to the long-term administration of ASA, DHI still decreased the gastric mucosal damage score in 52.0% from 2.50 to 1.20. DHI improved gastric mucus secretion, as well as decreased pepsin activity to maintain the integrity of gastric mucosal barrier (P<0.05). Furthermore, DHI recovered antioxidant activity which was impaired by ASA. In details, DHI decreased gastric mucosal ROS level, increased CAT, GSH-Px and SOD activity, and reduced MDA concentration (P<0.05). When ASA (71.9µM) was used in combination with DHI (23-fold dilution, presented in terms of concentrations of DSS, PA, SaD RA, SaB and SaA were 6.45-6.92, 1.10-1.14, 1.09-1.10, 0.86-0.90, 16.76-19.38 and 1.83-1.94µg/ml, respectively) in vitro, the inhibition rate of ASA increased from 38.6% (ASA alone) to 62.8% (ASA-DHI) on COX-1 and from 28.9% (ASA alone) to 38.8% (ASA-DHI) on COX-2 (P<0.05). DHI strengthened the inhibition activity of ASA on both COX-1 and COX-2, which showed that DHI alleviated ASA induced gastric mucosal damage but not antagonized anti-COX effect of ASA. CONCLUSIONS: Gastric protective benefits were clearly produced when DHI and ASA were used in combination, which provided rational guidance for clinical combined application of DHI and ASA.

[Application and prospect of model organism Drosophila melanogaster in activity evaluating and efficacy characterization of traditional Chinese medicine].

The common fruit fly, Drosophila melanogaster, is a well studied and tractable genetic model organism for exploring molecular mechanisms of human diseases in biological science. The considerable work of Drosophila has promoted the comprehension of relative protein expressions and signaling pathways associated with pathological and physiological activities. Meanwhile, various strains of transgenic drosophila with diverse genetic features have been established. These fly strains can be applied into bioactivity evaluation and drug screen as an emerging human disease model. The development of Chinese medicine has been seriously restricted by lacking of techniques and methods in activity evaluation. D. melanogaster, because of its many distinguishing features, such as rapid reproduction, short life cycle, rich strains, entirety action, highly correlated with human and other characteristics, has become a desirable choice to study Chinese medicine which has complicated composition. Here, progress of researches based on flies in disease models and their application in drug evaluating were reviewed, including aging, neurodegenerative diseases, metabolic disorders and diabetes, sleep disorder, intestinal immunity, reproduction, cancer and cardiac function.

[Study on evaluation system for gynecological disease model characterized by Qi stagnation and blood stasis syndrome with female SD rats].

OBJECTIVE: To establish an evaluation system for animal model with gynecological disease characterized by Qi stagnation and blood stasis syndrome, in order to disprove syndrome characteristics of the model by classic clinical prescriptions, and evaluate the specificity and reliability of the model with macroscopic biological signs and symptoms. METHOD: The model characterized by Qi stagnation and blood stasis syndrome was established by injecting adrenaline into female SD rats and conducting unpredictable chronic stimulus such as reversal of day and night, swimming in cold water, thermal stimulation in oven, noise and tail suspension for two weeks. They were also orally administered with Xiangfu Siwu Tang, Shaofu Zhuyu Tang and positive control drug aspirin in groups. A comprehensive evaluation was conducted for the model on the basis of haemorheology, four blood coagulation indexes, four diagnostic information (digital imaging of tongue, paw and tail, temperature, weight, ingestion, electrocardiograph, and open filed test), and syndrome rating. RESULT: Compared with the normal group, the model group showed obvious changes in haemorheology, four blood coagulation indexes, animal behavior, weight, ingestion, syndrome rating and heart rate. Their tongue and paw pictures were analyzed with Photoshop 7.0, showing significant difference in red, green and blue percentage composition from the normal group. Groups given aspirin and Xiangfu Siwu Tang showed notable changes in haemorheology, four blood coagulation indexes, animal behavior, weight, ingestion, heart rate, syndrome rating, and red, green and blue percentage composition in tongue and paw pictures, whereas the group given Shaofu Zhuyu Tang showed no remarkable improvement. CONCLUSION: The evaluation system for the animal model with gynecological disease characterized by Qi stagnation and blood stasis syndrome is established to provide reference for studies on the evaluation system for qi stagnation and blood stasis syndrome models.

[Effect of Yupingfeng San against OVA-induced allergic asthma in mice].

OBJECTIVE: To observe the effect of Yupingfeng San (YPFS) against OVA-induced allergic asthma in mice. METHOD: Mice were injected with OVA to establish the allergic asthma model. They were abdominally injected with 20 microg OVA on day 0 and 14, and inhaled aerosol 0.5% OVA solution for 20 min for seven days. The blank control group was administrated with equal volume of saline. YPFS groups with different doses were administrated intragastrically with YPFS every day, with the crude drug dosage of 3.25, 6.5, 13 g x kg(-1), respectively. The model group and control group were administrated with equal volume of saline. The positive control group was given intraperitoneally injected with 1 mg x kg(-1) DEX since aerosol inhalation. Blood was drawn after the last OVA aerosol inhalation to count the number of Eosnophils (Eos) in blood and detect IgE in serum; BALF was collected to count the number of cells and classify; right lung tissues were evenly grinded to detect cytokines IL-4 and IFN-gamma, and left upper lung lobes were collected for pathologic histology. RESULT: The level of Eos and IgE in serum increased significantly in the model group, and a large number of Eos were detected in BALF. Histopathological changes in lung showed bronchial serous exudation, tubular epithelial cells exfoliation, tube narrowing, widened alveolar septum, and bronchial periarterial lymphocytes infiltration. Homogenate of lung tissues showed increase of IL-4, and decrease in IFN-gamma/IL-4 ratio. YPFS groups with different doses displayed decrease of Eos in blood and BALF and IgE content in serum, and relief of pathologic changes in above models. Meanwhile, IL-4 content in homogenate of lung tissues decreased, with the increase in IFN-gamma/IL-4 ratio. CONCLUSION: YPFS shows the inhibitory effects on OVA-induced allergic asthma, involving down regulation of Eos and IgE levels in blood of asthma mice, and infiltration of inflammatory cells in lung tissues. Meanwhile, it can reduce IL-4 in lung homogenates, increase IFN-gamma/IL-4, and inhibits Th2 polarization.

UHPLC-MS simultaneous determination and pharmacokinetic study of three aromatic acids and one monoterpene in rat plasma after oral administration of Shaofu Zhuyu decoction.

We developed a sensitive and rapid method for determination of ferulic acid, caffeic acid, vanillic acid, and paeoniflorin in rat plasma based on ultra high performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS). The separation of the four compounds was carried out on an AcQuity UHPLC™ BEH C18 column using a mobile phase consisting of acetonitrile and water (containing 0.1% formic acid). Electrospray ionization in positive and negative ion mode and multiple reaction monitoring was used to identify and quantify active components. All calibration curves gave good linearity (r > 0.991) over the concentration range from 4.24-2875 ngmL(-1) for all components. The precision of the in vivo study was evaluated by intraday and interday assays and the percentages of RSD were all within 10.6%. The recovery ranged from 60.2 to 77.9%. The method was successfully applied to pharmacokinetic study of all three aromatic acids and one monoterpene in rat plasma. Furthermore, we compared the pharmacokinetics profile of the four compounds in normal and primary dysmenorrhea rats' plasma following oral administration of Shaofu Zhuyu decoction (SFZYD) and its ethanol supernatant extract (SFE).

Imbalance between Th17 and Treg cells may play an important role in the development of chronic unpredictable mild stress-induced depression in mice.

BACKGROUND: Recent data suggest that major depression is potentially associated with dysregulated cytokine production. However, the roles of T helper (Th) cells and their subsets in the development of depression still remain to be determined. The present study assessed changes in Th cell subsets and cytokines during the development of depression in a mouse model. METHODS: Chronic unpredictable mild stress (CUMS) was used to simulate depression behavior in mice. The open field test, sucrose preference, and ingestion were used as evaluative indicators of depressive behavior. During the CUMS protocol, on days 3, 7, 14, and 21, we assessed behavioral changes, cytokine levels in serum or stimulated (CD3/CD28) cell culture medium, and mRNA expression (ELISA, RT-PCR), regulatory T (Treg) and Th17 subsets in spleen (ex vivo, flow cytometry, RT-PCR), and CD3/rIL-23-stimulated Th17 cell proliferation (MTT assay). RESULTS: The results showed that in the depression model mice, IL-4 mRNA expression and serum levels increased on day 7, while no detectable change was observed in IFN-γ. Notably, a reduced proportion of Th17 cells with decreased proliferation capacity was observed at later stages, in parallel with a decline in serum IL-23 levels. In contrast, an increased Treg cell proportion and increased Foxp3 mRNA expression were observed in the mid-stages. Correlation analysis showed that the proportion of Tregs was correlated negatively with sucrose preference, while the proliferation of Th17 cells was notably correlated positively with sucrose preference. Also, an increased TGF-ß level was detected in serum and was believed to be a key factor responsible for the imbalance between Th17 and Treg cells. Furthermore, the sucrose preference in TGF-ß type I receptor blockade mice increased considerably, compared with CUMS mice. CONCLUSION: These results indicate that in CUMS-induced depression, behavioral changes may closely correlate with the imbalance between Th17 and Treg cell subsets, and TGF-ß may be a key regulatory cytokine.

A method of hepatocyte extraction conjugated with HPLC is established for screening potential active components in Chinese medicines--probing Herba Artemisiae Scopariae as an exemplifying approach.

In order to establish an effective and quick method for screening potential bioactive compounds in Traditional Chinese Medicines (TCMs), hepatocytes were employed for extracting either bifendate, a clinical medicine for liver diseases, or chemicals in Herba Artemisiae Scopariae (A. Scopariae), a commonly used traditional Chinese medicine for remedying liver diseases such as hepatitis induced by viruses, chemicals or alcohol. After hepatocyte extraction the compounds were analyzed by HPLC, therefore this method was referrred to as hepatocyte extraction conjugated with HPLC (HE-HPLC). In the first part of this study, HE-HPLC showed that bifendate was extracted by hepatocytes and detected by HPLC-DAD which indicated the feasibility of this method. Then in the second part of the study, the potential active components in the A. scopariae extract were studied using HE-HPLC. Six chemicals in the A. scopariae extract, which could bind to hepatocytes in vitro, were detected by HPLC-DAD and three were identified as 7-hydroxy-coumarin (7-OH-C), capillartemisin A and 7-methoxy-coumarin, respectively. In vitro assays showed that 7-OH-C protected HL-7702 hepatocytes from H2O2 injury. The results indicated that these compounds could be extracted by hepatocytes, could be detected by HPLC and more importantly were bioactive. It is suggested that HE-HPLC is a useful method for screening potent active components in Chinese medicines used to treat liver diseases.