Mineral crude drug mirabilite (Mangxiao) inhibits the occurrence of colorectal cancer by regulating the Lactobacillus-bile acid-intestinal farnesoid X receptor axis based on multiomics integration analysis.

Mineral crude drug has revolutionized the treatment landscape in precision oncology niche that leads to the improvement in therapeutic efficiency on various tumor subtypes. Mangxiao (MX), a mineral crude drug in traditional Chinese medicine, has been used for treating gastrointestinal diseases for thousands of years. However, the action mechanisms are still ambiguous. Here, we attempt to explore inhibitory roles and associated pharmacological mechanisms of MX upon colorectal cancer (CRC) in APCMin/+ male mice by integrating metabolomics, 16S rDNA sequencing analyses, and metagenomic-based microbiota analysis. We found that MX can significantly inhibit the occurrence of CRC through the regulation of the dysregulated gut microbe metabolism. Furthermore, the correlation analysis of metabolomes and 16S rDNA revealed that MX could restore the disorders of gut microbes by specifically enriching the abundance of Lactobacilli to improve bile acid metabolism, which further activated the farnesoid X receptor (FXR) in CRC mice, then the improvement of gut dysbiosis could inhibit the development of CRC. Collectively, our effort confirmed MX has the capacity to intervene the development of CRC and further discovered that it targets Lactobacillus-bile acid-intestinal FXR axis, which can be regarded as a candidate medicine for future drug discovery and development against CRC.

Clinical Study of Percutaneous Endoscopic Large-channel Fusion and Transforaminal Lumbar Interbody Fusion in the Treatment of Degenerative Lumbar Spinal Stenosis.

Context: Degenerative changes in the lumbar spine more commonly cause spinal stenosis and with the aging of society, its incidence is on the rise. Endoscopic spinal surgery is a minimally invasive technique for decompression. The efficacy of percutaneous, endoscopic, large-channel fusion and transforaminal lumbar interbody fusion (TLIF) need confirmation by more studies. Objective: The study intended to investigate the clinical efficacy of percutaneous endoscopic large-channel fusion and TLIF in the treatment of degenerative lumbar spinal stenosis, to find the best treatment plan. Design: The research team performed a retrospective study. Setting: The study took place at Nanjing Lishui People's Hospital in Nanjing, Jiangsu Province, PR China. Participants: Participants were 100 patients with degenerative, lumbar, spinal stenosis who had been admitted to the hospital between October 2018 and October 2022. Intervention: The research team randomly divided participants into an intervention group and a control group, with 50 participants in each group. The intervention group received percutaneous, endoscopic, large-channel fusion and internal fixation, and the control group received foraminal, lumbar, interbody fusion. Outcome Measures: The research team measured: (1) perioperative indexes, (2) clinical efficacy at a postoperative follow-up at 6 months postintervention, (3) indexes for inflammatory responses at baseline and postintervention, (4) postoperative pain at baseline and at months 3 and 6 postintervention using a visual analog scale (VAS), (6) lumbar function at baseline and months 3 and 6 postintervention using the Oswestry Disability Index (ODI) and the Japanese Orthopedic Association (JOA) scale, and (7) complications. Results: Compared with the control group, the intervention group's perioperatively related and inflammatory-response indexes were significantly better: (1) amount of bleeding- 112.67 ± 17.38 for the control group and 78.62 ± 10.52 for the intervention group (P = .002); (2) volume of drainage-79.63 ± 14.21 for the control group and 52.18 ± 8.21 for the intervention group (P = .001); (3) ESR at baseline and postintervention-22.41 ± 5.62 and 15.18 ± 5.26, respectively, for the control group and 22.58 ± 5.82 and 10.54 ± 3.18, respectively, for the intervention group, with P = .013 postintervention; and (4) CRP at baseline and postintervention-17.42 ± 3.52 and 13.98 ± 3.65 for the control group, respectively, and 18.65 ± 3.78 and 10.14 ± 2.78 for the intervention group, with P = .008 postintervention; Also, compared to the control group, the intervention group's: (1) total effective rate was significantly higher (P = .018); (2) incidence of postoperative complications was significantly lower (P = .006); (3) VAS pain score was significantly lower at months 3 and 6, with P = .028 and P = .021, respectively; (4) Oswestry Disability Index (ODI) function score was significantly lower at months 3 and 6, with P = .016 and P = .014, respectively; and (5) postoperative JOA function score was significantly higher at months 3 and 6, with P = .011 and P = .007, respectively. Conclusions: Both percutaneous, endoscopic, large-channel fusion and TLIF had good therapeutic effects in the treatment of degenerative lumbar spinal stenosis. However, compared with the latter, the former was more effective, with better comprehensive efficacy and more obvious benefits for patients, so it's worthy of clinical promotion and use.

Application of preparative high-speed countercurrent chromatography for the separation and purification of three flavonoids from Oroxylum indicum.

An efficient method was established by high-speed countercurrent chromatography (HSCCC) for the separation and purification of three flavonoids from Oroxylum indicum. Optimized by single-factor and orthogonal experiments, the optimal extraction conditions were an extraction temperature of 50°C, a solid-to-liquid ratio of 1:50 (g/ml), an ethanol concentration of 75% and an extraction time of 45 min. Using a two-phase solvent system composed of chloroform-methanol-water (6:10:5, v/v/v), the preparative separation was successfully performed by HSCCC in head-to-tail elution mode. Totals of 12.63 mg of oroxin A at a purity of 97.61% with 96.46% recovery, 10.96 mg oroxin B at a purity of 98.32% with 98.81% recovery, and 9.34 mg baicalein at a purity of 98.64% with 97.87% recovery were obtained in one-step separation from 200 mg crude extract. Their chemical structures were confirmed by melting points, HPLC, UV, FTIR, MS, 1 H and 13 C NMR data. Furthermore, they were efficient scavengers of 1,1-diphenyl-2-picrylhydrazyl and hydroxyl free radicals in a concentration-dependent manner.

Optimization, extraction, and purification of three bioactive compounds from Entada phaseoloides by high-speed countercurrent chromatography.

The objective of this paper was to develop a preparative method for the separation and purification of phaseoloidin, entadamide A, and entadamide A-ß-D-glucopyranoside from the crude extract of Entada phaseoloides by high-speed countercurrent chromatography (HSCCC) for the first time. Optimized by orthogonal experiments, the extraction conditions were extraction temperature of 65°C, solid-to-liquid ratio of 1:15 (g/mL), ethanol concentration of 40%, and extraction time of 2.5 h. Using n-butanol-acetic acid-water (4:1:5, v/v/v) as the two-phase solvent system, 38.79 mg phaseoloidin (the purity was 99.3% with a recovery of 98.1%), 34.85 mg entadamide A (the purity was 96.4% with a recovery of 98.5%), and 33.97 mg entadamide A-ß-D-glucopyranoside (the purity was 98.6% with a recovery of 97.7%) were obtained from 500 mg crude extract by HSCCC in head-to-tail elution mode. The retention ratio of stationary phase was 51.0%. According to the antioxidant activity assays, phaseoloidin, entadamide A, and entadamide A-ß-D-glucopyranoside had certain scavenging abilities on 1,1-diphenyl-2-picrylhydrazyl free radicals and hydroxyl free radicals.

Preparative Isolation and Purification of Calycosin and Formononetin from Astragali Radix using Hydrolytic Extraction Combined with High Speed Countercurrent Chromatography.

Calycosin and formononetin were efficiently extracted from Astragali Radix and purified by high-speed countercurrent chromatography. Calycosin and formononetin could be hydrolyzed from calycosin-7-glucoside and ononin, respectively. The best extraction conditions were realized by single factor and orthogonal experiments, which were 100% ethanol, 2.5 mol/L hydrochloric acid, 1:40 ratio of solid to liquid, extracted 2 h and one time. The two-phase solvent system of n-hexane-ethyl acetate-ethanol-water (3:5:3:5, v/v) was selected for the purification of calycosin, and 1.3 mg calycosin (the purity was 95.8% and the recovery was 85.9%) was obtained from 264.9-mg crude extraction. The two-phase solvent system of n-hexane-ethyl acetate-ethanol-water (4:5:4:5, v/v) was selected for the purification of formononetin, and 2.0 mg formononetin (the purity was 98.9% and the recovery was 84.4%) was obtained from 248.9-mg crude extraction. Their structures were identified by HPLC, melting points, UV, FTIR, ESI-MS, 1H NMR and 13C NMR spectrum. According to the antioxidant activity assay, the scavenging abilities of calycosin to 1,1-diphenyl-2-picrylhydrazyl and hydroxyl free radicals (·OH) were stronger. The scavenging effect of formononetin was not demonstrated.

Functional metabolomics innovates therapeutic discovery of traditional Chinese medicine derived functional compounds.

Traditional Chinese medicines (TCMs) produce chemically diverse functional compounds that are importantly chemical resource for facilitating new drug discovery and development against a diversity of diseases. However, modern exploration of TCM derived functional compounds is significantly hindered by the inefficient elucidation of pharmacological functions over past decades, because conventional research methods are incapable of efficiently elucidating therapeutic potential of TCM conferred by multiple functional compounds. Functional metabolomics has the priority-capacity to characterize systems therapeutic actions of TCM by precisely capturing molecular interactions between disease response metabolite biomarkers (DRMB) and functional compounds (secondary metabolites), which underline pharmacological efficiency and associated therapeutic mechanisms. In this critical review, we innovatively summarize systems therapeutic feature of TCM derived functional compounds from a functional-metabolism perspective, then systems metabolic targets (SMT) identified by functional metabolomics method are strategically proposed to better understanding of therapeutic discovery of TCM derived functional compounds. In addition, we propose the perspective strategy as Spatial Temporal Operative Real Metabolomics (STORM) to considerably improve analytical capacity of functional metabolomics method by selectively incorporating the cutting edge technologies of mass spectrometry imaging, isotope-metabolic fluxomics, synthetic and biosynthetic chemistry, which could considerably enhance the precision and resolution of elucidating pharmacological efficiency and associated therapeutic mechanisms of TCM derived functional compounds. Collectively, such critical review is expected to provide novel perspective-strategy that could significantly improve modern exploration and exploitation of TCM derived functional compounds that further promote new drug discovery and development against the complex diseases.

Acute cognitive disorder as the initial manifestation of nitrous oxide abusing: a case report.

Nitrous oxide (N2O) is a colorless, odorless gas used as an anesthetic and analgesic. It is also abused as a recreational drug, and such abuse is associated with neurological disorders and psychiatric complications such as myelopathy and, rarely, cognitive impairment. Its abuse has not been associated with acute cognitive decline. Here, we report a young girl who presented with acute cognitive impairment after excessive recreational inhalation of nitrous oxide and who recovered completely after vitamin B12 supplementation. We conclude that nitrous oxide abuse can cause acute cognitive impairment, and that this diagnosis should be considered if a patient initially presents with acute cognitive decline.

Berberine improves colitis by triggering AhR activation by microbial tryptophan catabolites.

Inflammatory bowel diseases (IBD) are kind of recurrent inflammatory issues that occur in the gastrointestinal tract, and currently clinical treatment is still unideal due to the complex pathogenesis of IBD. Basically, gut barrier dysfunction is triggered by gut microbiota dysbiosis that is closely associated with the development of IBD, we thus investigated the therapeutic capacity of berberine (BBR) to improve the dysregulated gut microbiota, against IBD in rats, using a combinational strategy of targeted metabolomics and 16 s rDNA amplicon sequencing technology. Expectedly, our data revealed that BBR administration could greatly improve the pathological phenotype, gut barrier disruption, and the colon inflammation in rats with dextran sulfate sodium (DSS)-induced colitis. In addition, 16S rDNA-based microbiota analysis demonstrated that BBR could alleviate gut dysbiosis in rats. Furthermore, our targeted metabolomics analysis illustrated that the levels of microbial tryptophan catabolites in the gastrointestinal tract were significantly changed during the development of the colitis in rats, and BBR treatment can significantly restore such changes of the tryptophan catabolites accordingly. At last, our in vitro mechanism exploration was implemented with a Caco-2 cell monolayer model, which verified that the modulation of the dysregulated gut microbiota to change microbial metabolites coordinated the improvement effect of BBR on gut barrier disruption in the colitis, and we also confirmed that the activation of AhR induced by microbial metabolites is indispensable to the improvement of gut barrier disruption by BBR. Collectively, BBR has the capacity to treat DSS-induced colitis in rats through the regulation of gut microbiota associated tryptophan metabolite to activate AhR, which can greatly improve the disrupted gut barrier function. Importantly, our finding elucidated a novel mechanism of BBR to improve gut barrier function, which holds the expected capacity to promote the BBR derived drug discovery and development against the colitis in clinic setting.

Precision-characterization and quantitative determination of main compounds in Si-Ni-San with UHPLC-MS/MS based targeted-profiling method.

Mass spectrometry based precision identification of natural products requires the validation of the reference compounds. This study attempted to develop a LC-QTOF MS combined with LC-TQ MS method to precisely characterize the chemicals of Si-Ni-San (SNS), a classic traditional Chinese medicine formula, which is composed of four medicinal plants, and widely used for the treatments of liver disorders. 74 compounds in SNS were provisionally identified by acquiring MS spectra and MS/MS spectra of the possible chemical features, as well as retrieving small-molecule database. By comparing with the accurate MS/MS spectra of reference compounds, 37 compounds in SNS were precisely identified for the first time. In addition, our effort also successfully assigned the origin of each identified compounds against four medicinal plants. Furthermore, we developed a UHPLC-TQ MS based quantitative-profiling method for simultaneous determination of 37 targeted compounds in the different extracts of the raw SNS and commercial lyophilized powders, enabling to facilitate overall quality control of SNS and associated commercial products. Collectively, our finding precisely characterized the main chemicals in SNS, which also provides a new strategy with LC-MS/MS based chemical profiling to precisely identify a diversity of chemicals in Chinese medicinal plants and associated formulae.

Preparative Separation and Purification of Liquiritin and Glycyrrhizic Acid from Glycyrrhiza uralensis Fisch by High-Speed Countercurrent Chromatography.

The technique of high-speed countercurrent chromatography (HSCCC) was applied to the preparative isolation and purification of liquiritin and glycyrrhizic acid from a crude extract of Glycyrrhiza uralensis Fisch for the first time. Using single factor and orthogonal design experiments, the best extraction conditions were 70% ethanol, 1:25 ratio of solid-to-liquid (w/v) and extracted 1.5 h at 80°C. The contents of liquiritin and glycyrrhizic acid in the crude extract were 1.3 and 5.3%, respectively. Using the two-phase solvent system of ethyl acetate-methanol-water (5:2:5, v/v), 6.0 mg liquiritin (the purity was 96.7%, and the recovery was 89.3%), and 20.5 mg glycyrrhizic acid (the purity was 98.9%, and the recovery was 77.1%) were obtained from 500 mg crude extraction by HSCCC, respectively. The retention rate of stationary phase was 51.0%. Their structures were identified by high-performance liquid chromatography, melting points, ultraviolet radiation, Fourier transform infrared (FTIR), Electrospray ionization mass spectrometry (ESI-MS), 1H Nuclear Magnetic Resonance (NMR) and 13C NMR spectra. The scavenging abilities of glycyrrhizic acid to 1,1-diphenyl-2-picrylhydrazyl and hydroxyl free radicals were stronger than those of liquiritin.

Omics strategies decipher therapeutic discoveries of traditional Chinese medicine against different diseases at multiple layers molecular-level.

Traditional Chinese medicine (TCM) has been broadly used for the personalized treatment of many diseases in China for thousands of years. In the past century, TCM was also introduced to other Asian countries and even the Western world. Increasing evidence has shown that TCM has the capacity to treat numerous complex diseases in the clinic, such as cardiovascular diseases (CVDs), infectious diseases, metabolic diseases, and neurodegenerative diseases. However, the earlier lack of analytical strategies to annotate the chemical complexity has severely impeded the modern study and translational application of TCM. This critical review aims to explore and exploit applications of systems biology-driven omics methods in TCM against a diversity of diseases, toward the specific use of TCM to treat patients with different diseases. Such effort shall enhance the applicability of systems biology-driven omics strategies in deciphering the mechanisms by which TCM treats different diseases and may lead to the discovery of new therapeutic directions. In addition, we proposed the possible strategies to innovate the applicable pattern of omics technologies in TCM niches, such as precision-modification metabolomics and chinmedomics methods, allowing to unveil the complexity of TCM, which must enable TCM to serve better for the population-health. Taken together, this review eventually shall highlight the core value of omics technologies in innovating TCM to combat the diseases in a new horizon.

Preparative separation of liquiritigenin and glycyrrhetic acid from Glycyrrhiza uralensis Fisch using hydrolytic extraction combined with high-speed countercurrent chromatography.

The objective of this paper was to develop a preparative method for the isolation and purification of liquiritigenin and glycyrrhetic acid from Glycyrrhiza uralensis Fisch using hydrolytic extraction combined with high-speed countercurrent chromatography (HSCCC). Liquiritigenin and glycyrrhetic acid were well hydrolyzed from liquiritin and glycyrrhizic acid by hydrochloric acid, respectively. The optimal extraction conditions were obtained by single-factor and orthogonal experiments, which were 100% ethanol, 1.5 mol/L hydrochloric acid, 1:25 ratio of solid to liquid, and extracted 2 h for one time. Using the two-phase solvent system of n-hexane-ethyl acetate-methanol-water (4:5:4:5, v/v), 2.1 mg liquiritigenin (the purity was 96.5% with a recovery of 87.6%) and 12.3 mg glycyrrhetic acid (the purity was 97.1% with a recovery of 74.4%) were obtained from 315-mg crude extraction by HSCCC. The retention ratio of stationary phase was 47.2%. Their structures were identified by HPLC, melting points, UV, Fourier-transform infrared, Electrospray ionization-MS, 1 H nuclear magnetic resonance (NMR), and 13 C NMR spectra. According to the antioxidant activity assays, liquiritigenin and glycyrrhetic acid had some scavenging abilities on 1,1-diphenyl-2-picrylhydrazyl free radicals; liquiritigenin had stronger scavenging ability on hydroxyl radicals.

Preparative separation of quercetin, ombuin and kaempferide from Gynostemma pentaphyllum by high-speed countercurrent chromatography.

An efficient method for the preparative separation of quercetin, ombuin and kaempferide from Gynostemma pentaphyllum by high-speed countercurrent chromatography (HSCCC) was successfully developed for the first time. The extraction conditions were optimized by an orthogonal array design L9 (34), while quercetin, ombuin and kaempferide were efficiently released from rutin, ombuoside and kaempferide glucoside by hydrochloric acid hydrolysis, respectively. Quercetin was purified by HSCCC with two-phase solvent system composed of n-hexane-ethyl acetate-methanol-water (4:5:4:5, v/v) in a single run. From 164.7 mg of the crude extract, 90.5 mg quercetin at 99.23% purity was obtained. Ombuin and kaempferide were purified by HSCCC with two-phase solvent system composed of n-hexane-ethyl acetate-methanol-water (8:2:5:5, v/v) in a single run. From 50 mg of the crude extract, 0.4 mg of ombuin at 95.46% purity and 1.1 mg of kaempferide at 98.78% purity were obtained, respectively. Their structures were identified by ultraviolet, Fourier transform infrared, electrospray ionization mass spectrometry (ESI-MS), 1H nuclear magnetic resonance (NMR) and 13C NMR spectra. The purified quercetin had the strongest 1,1-diphenyl-2-picrylhydrazyl and hydroxyl free radical scavenging activities.

Separation and Purification of Ombuoside from Gynostemma Pentaphyllum by Microwave-Assisted Extraction Coupled with High-Speed Counter-current Chromatography.

A rapid and efficient method for the separation and purification of ombuoside from Gynostemma pentaphyllum by microwave-assisted extraction coupled with high-speed counter-current chromatography (HSCCC) was successfully developed. Using an orthogonal array design L9 (34), the extraction conditions, including microwave power, irradiation time, solid-to-liquid ratio and extraction times, were optimized. Ombuoside was isolated and purified from the crude extraction by HSCCC with two-phase solvent system composed of n-hexane:ethyl acetate:ethanol:water (5:6:5:5, v/v) in a single run. A 210 mg quantity of the crude extract containing 2.16% ombuoside was loaded, yielding 3.9 mg of ombuoside at 96.7% purity. The chemical structure of ombuoside was determined by comparison with the high-performance liquid chromatography retention time of standard substance as well as UV, FT-IR, ESI-MS, 1H NMR and 13C NMR spectra. The purified ombuoside had strong 1,1-diphenyl-2-picrylhydrazyl and hydroxyl free radical scavenging activities.

Microwave-Assisted Extraction and Purification of Arctiin and Arctigenin from Fructus Arctii by High-Speed Countercurrent Chromatography.

An efficient method for the rapid extraction, separation and purification of bioactive lignans, arctiin and arctigenin, from Fructus arctii by microwave-assisted extraction coupled with high-speed countercurrent chromatography was developed. The optimal extraction conditions of arctiin and arctigenin were evaluated by orthogonal array. Arctigenin could be converted from arctiin by hydrochloric acid hydrolysis. The separations were performed at a preparative scale with two-phase solvents composed of ethyl acetate-ethanol-water (5 : 1 : 5, v/v/v) for arctiin, and n-hexane-ethyl acetate-ethanol-water (4 : 4 : 3 : 4, v/v/v/v) for arctigenin. From 500 mg of crude extract sample, 122.3 mg of arctiin and 45.7 mg of arctigenin were obtained with the purity of 98.46 and 96.57%, and the recovery of 94.3 and 81.6%, respectively. Their structures were determined by comparison with the high-performance liquid chromatography retention time of standard substance as well as UV, FT-IR, electrospray ion source (ESI)-MS, (1)H-NMR and (13)C-NMR spectrum. According to the antioxidant activity assay, arctigenin had stronger 1,1-diphenyl-2-picrylhydrazyl free radicals scavenging activity.

Preparation and antibacterial activity studies of degraded polysaccharide selenide from Enteromorpha prolifera.

Polysaccharide extracted from Enteromorpha prolifera possessed excellent biological activities, but its molecular weight was greatly high which influenced the activity. Organic Se had higher biological activities and was safer than inorganic Se species. In the present study, degraded polysaccharide selenide (Se-LEP) was synthesized from sodium selenite and degraded polysaccharide (LEP) with the catalysis of nitric acid. The preparation conditions of LEP and Se-LEP were optimized by orthogonal experiments. The selenite ester group was formed, and the selenium content was 1335.27 µg/g. The thermal stability of Se-LEP decreased. LEP had less inhibitory effects on bacteria and plant pathogenic fungi. Se-LEP had stronger inhibitory effect on Eschetichia coli, and weaker inhibitory effect on Staphylococcus aureus than polysaccharide selenide (Se-EP). Se-LEP also had better inhibitory effects on plant pathogenic fungi.

Preparative isolation and purification of indigo and indirubin from Folium isatidis by high-speed counter-current chromatography.

INTRODUCTION: Indigo and indirubin are the main active ingredients found in traditional Chinese herbal medicine Folium isatidis. An effective method for the isolation and purification of indigo and indirubin from Folium isatidis is needed. Compared with the conventional column chromatographic techniques, high-speed counter-current chromatography (HSCCC) is a suitable alternative for the enrichment and purification of these target compounds, and eliminates the complications resulting from a solid support matrix. OBJECTIVE: To develop a reliable HSCCC method for isolation and identification of indigo and indirubin in a one-step separation from Folium isatidis. METHODOLOGY: The optimum extracting conditions of indigo and indirubin from Folium isatidis were investigated by orthogonal test L(16) (4(5)). The target compounds were isolated and purified with a solvent system of n-hexane:ethyl acetate:ethanol:water (1:1:1:1, v/v) and the lower phase was used as the mobile phase in the head-to-tail elution mode. The purities of target compounds were tested by HPLC and their structures were identified by UV, IR, electrospray ion source (ESI)-MS, (1) H-NMR and (13) C-NMR analyses. RESULTS: From 165 mg of the crude extract, 5.65 mg of indigo and 1.00 mg of indirubin were obtained by HPLC analysis with purities of 98.4% and 99.0% respectively, and their mean recoveries were 91.0% and 90.7%, respectively. CONCLUSION: The HSCCC method is effective for the preparative separation and purification of indigo and indirubin in a one-step separation from Folium isatidis.

New insights into glutamate ototoxicity in cochlear hair cells and spiral ganglion neurons.

CONCLUSION: Excess glutamate (Glu) exposure (20 mM) in the cochlear perilymph affects the physiological function of outer hair cells (OHCs) within a 2 h period and induces apoptosis in the modiolus spiral ganglion neurons (SGNs) in an apoptosis-inducing factor (AIF)-dependent manner. OBJECTIVES: To determine whether high-dose Glu affects the function of OHCs and whether it induces AIF- and caspase-3-dependent apoptosis in the cochlear SGNs. METHODS: Perilymphatic perfusions of Glu (20 mM) and artificial perilymph (AP) solutions were performed in adult guinea pig cochleae. Both cochlear microphonics (CM) and electrical auditory brainstem response (eABR) were measured before and 2 h after perfusions. The hair cell morphologies were examined using transmission electron microscopy. The expression of two apoptotic indicators, AIF and caspase-3, was examined 8 h after perfusions. RESULTS: In contrast to AP perfusions, the perfusion of 20 mM Glu caused significant reduction in the CM and eABR amplitudes. Inner hair cells (IHCs) after Glu perfusion were deformed and exhibited vacuolization in the postsynaptic region, whereas the OHC system appeared unaffected. AIF expression was detected in the nuclei of SGNs 8 h after Glu exposure, but the expression of caspase-3 was not shown in any cochlear tissues.

Comparative study on the protective effects of Yinchenhao Decoction against liver injury induced by alpha-naphthylisothiocyanate and carbon tetrachloride.

OBJECTIVE: To optimize the animal model of liver injury that can properly represent the pathological characteristics of dampness-heat jaundice syndrome of traditional Chinese medicine. METHODS: The liver injury in the model rat was induced by alpha-naphthylisothiocyanate (ANIT) and carbon tetrachloride (CCl(4) ) respectively, and the effects of Yinchenhao Decoction (, YCHD), a proved effective Chinese medical formula for treating the dampness-heat jaundice syndrome in clinic, on the two liver injury models were evaluated by analyzing the serum level of alanine aminotransferase (ALT), asparate aminotransferase (AST), alkaline phosphatase (ALP), malondialchehyche (MDA), total bilirubin (T-BIL), superoxide dismutase (SOD), glutathione peroxidase (GSH-PX) as well as the ratio of liver weight to body weight. The experimental data were analyzed by principal component analytical method of pattern recognition. RESULTS: The ratio of liver weight to body weight was significantly elevated in the ANIT and CCl(4) groups when compared with that in the normal control (P<0.01). The contents of ALT and T-BIL were significantly higher in the ANIT group than in the normal control (P<0.05,P<0.01), and the levels of AST, ALT and ALP were significantly elevated in CCl(4) group relative to those in the normal control P<0.01). In the YCHD group, the increase in AST, ALT and ALP levels was significantly reduced (P<0.05, P<0.01), but with no significant increase in serum T-BIL. In the CCl(4) intoxicated group, the MDA content was significantly increased and SOD, GSH-PX activities decreased significantly compared with those in the normal control group, respectively (P<0.01). The increase in MDA induced by CCl(4) was significantly reduced by YCHD P<0.05). CONCLUSION: YCHD showed significant effects on preventing liver injury progression induced by CCl(4), and the closest or most suitable animal model for damp-heat jaundice syndrome may be the one induced by CCl(4).

Purification of paeoniflorin from Paeonia lactiflora Pall. by high-speed counter-current chromatography.

High-speed counter-current chromatography was successfully used for the first time for the preparative separation and purification of paeoniflorin from the Chinese medicinal plant Paeonia lactiflora Pall. using a two-phase solvent system composed of n-butanol-ethyl acetate-water (1:4:5, v/v) in a single run. From 160 mg of the crude sample containing 22.0% paeoniflorin, 33.2 mg of paeoniflorin was yielded at 98.2% purity as determined by HPLC analysis. The recovery of paeoniflorin was 94.3%.