Network pharmacology, molecular docking, combined with experimental verification to explore the role and mechanism of shizhifang decoction in the treatment of hyperuricemia.

Ethnopharmacological relevance: Shizhifang Decoction, a traditional Chinese medicine prescription formulated by Professor Zheng Pingdong of Shuguang Hospital, has been widely utilized in clinical settings for the treatment of hyperuricemia due to its proven safety and efficacy. Objective: In this study, we used network pharmacology, molecular docking technology, and experimental validation to elucidate the therapeutic effects and underlying mechanisms of Shizhifang Decoction in managing hyperuricemia. Methods: Quality control and component identification of the freeze-dried powder of Shizhifang Decoction were conducted using ultra-high performance liquid chromatography-tandem quadrupole time-of-flight mass spectrometry. Active ingredients and their corresponding targets were obtained from Traditional Chinese Medicine Systems Pharmacology, Traditional Chinese Medicine Information Database, The Encyclopedia of Traditional Chinese Medicine, and other databases. Disease-related targets for hyperuricemia were collected from GeneCards and DisGeNET databases. The Venny platform is used to screen common targets for drug active ingredients and diseases. Subsequently, we constructed an active component-target-disease interaction network using the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) database, create a component disease common target network using Cytoscape 3.9.1 software, from which core targets were selected. Import common targets into the Database for Annotation, Visualization and Integrated Discovery (DAVID) for Gene Ontology enrichment and Kyoto Encyclopedia of Genes and Genomes pathway analysis. Molecular docking was then conducted to validate the binding capacity of key active ingredients and their associated targets in Shizhifang Decoction. The theoretical predictions were further confirmed through in vitro and in vivo experiments. Result: A total of 35 active ingredients and 597 action targets were identified, resulting in 890 disease-related targets for hyperuricemia. After comprehensive analysis, 99 common targets were determined. Protein-protein interaction network analysis revealed crucial relationships between these targets and hyperuricemia. Among them, 12 core targets (CASP3, IL1B, IL6, TNF, TP53, GAPDH, PTGS2, MYC, INS, VEGFA, ESR1, PPARG) were identified. Gene Ontology enrichment analysis demonstrated significant associations with the regulation of inflammatory response, cell apoptosis, and the positive regulation of extracellular regulated protein kinases 1 and extracellular regulated protein kinases 2 cascades. Kyoto Encyclopedia of Genes and Genomes pathway analysis highlighted inflammation and apoptosis-related pathways as critical mediators of Shizhifang Decoction's effects on hyperuricemia. Molecular docking studies further supported the interactions between apoptosis-related proteins and active ingredients in the extracellular regulated protein kinases 1/2 signaling pathway. In vitro experiments confirmed the downregulation of apoptosis-related proteins (caspase-3, Bax, Bcl-2) and the inhibition of the extracellular regulated protein kinases 1/2 signaling pathway by Shizhifang Decoction. These findings were also validated in animal models, demonstrating the potential of Shizhifang Decoction to mitigate renal injury induced by hyperuricemia through extracellular regulated protein kinases 1/2-mediated inhibition of renal tubular epithelial cell apoptosis. Conclusion: Our study provides valuable insights into the main mechanism by which Shizhifang Decoction ameliorates hyperuricemia. By targeting the ERK1/2 signaling pathway and modulating cell apoptosis, Shizhifang Decoction exhibits promising therapeutic potential for the treatment of hyperuricemia. These findings support the continued exploration and development of Shizhifang Decoction as a potential herbal remedy for hyperuricemia management.

Ginkgo biloba extract (EGb-761) confers neuroprotection against ischemic stroke by augmenting autophagic/lysosomal signaling pathway.

Ginkgo biloba extract (EGb-761) is well-recognized to have neuroprotective properties. Meanwhile, autophagy machinery is extensively involved in the pathophysiological processes of ischemic stroke. The EGb-761 is widely used in the clinical treatment of stroke patients. However, its neuroprotective mechanisms against ischemic stroke are still not fully understood. The present study was conducted to uncover whether the pharmacological effects of EGb-761 can be executed by modulation of the autophagic/lysosomal signaling axis. A Sprague-Dawley rat model of ischemic stroke was established by middle cerebral artery occlusion (MCAO) for 90 min, followed by reperfusion. The EGb-761 was then administered to the MCAO rats once daily for a total of 7 days. Thereafter, the penumbral tissues were acquired to detect proteins involved in the autophagic/lysosomal pathway including Beclin1, LC-3, SQSTM1/p62, ubiquitin, cathepsin B, and cathepsin D by western blot and immunofluorescence, respectively. Subsequently, the therapeutic outcomes were evaluated by measuring the infarct volume, neurological deficits, and neuron survival. The results showed that the autophagic activities of Beclin1 and LC3-II in neurons were markedly promoted by 7 days of EGb-761 therapy. Meanwhile, the autophagic cargoes of insoluble p62 and ubiquitinated proteins were effectively degraded by EGb-761-augmented lysosomal activity of cathepsin B and cathepsin D. Moreover, the infarction size, neurological deficiencies, and neuron death were also substantially attenuated by EGb-761 therapy. Taken together, our study suggests that EGb-761 exerts a neuroprotective effect against ischemic stroke by promoting autophagic/lysosomal signaling in neurons at the penumbra. Thus, it might be a new therapeutic target for treating ischemic stroke.

Shizhifang ameliorates pyroptosis of renal tubular epithelial cells in hyperuricemia through inhibiting NLRP3 inflammasome.

ETHNOPHARMACOLOGICAL RELEVANCE: Traditional Chinese Medicine (TCM) Compound Shizhifang (SZF), consisting of the seeds of four Chinese herbs, has been used in Shanghai Shuguang Hospital in China for more than 20 years and has proven its clinical safety and efficacy in lowering uric acid and protecting kidney function. AIM OF THE STUDY: Hyperuricemia (HUA)-induced pyroptosis of renal tubular epithelial cells serves as a significant cause of tubular damage. SZF proves to be effective in alleviating renal tubular injury and inflammation infiltration of HUA. However, the inhibiting effect of SZF on pyroptosis in HUA still remains elusive. This study aims to verify whether SZF could ameliorate pyroptosis in tubular cells induced by uric acid (UA). MATERIALS AND METHODS: Quality control analysis and chemical and metabolic identification for SZF and SZF drug serum were performed by using UPLC-Q-TOF-MS. In vitro, human renal tubular epithelial cells (HK-2) stimulated by UA were treated with SZF or NLRP3 inhibitor (MCC950). HUA mouse models were induced by intraperitoneal injection of potassium oxonate (PO). Mice were treated with SZF, allopurinol or MCC950. We focused on evaluated the effect of SZF on the NLRP3/Caspase-1/GSDMD pathway, renal function, pathologic structure and inflammation. RESULTS: SZF significantly restrained the activation of the NLRP3/Caspase-1/GSDMD pathway in vitro and in vivo induced by UA. SZF was better than allopurinol and MCC950 in reducing pro-inflammatory cytokine levels, attenuating tubular inflammatory injury, inhibiting interstitial fibrosis and tubular dilation, maintaining tubular epithelial cell function, and protecting kidney. Furthermore, 49 chemical compounds of SZF and 30 metabolites in serum after oral administration were identified. CONCLUSIONS: SZF inhibits UA-induced renal tubular epithelial cell pyroptosis via by targeting NLRP3 to inhibit tubular inflammatory and prevent the progression of HUA-induced renal injury effectively.

Image fusion technique for target volume delineation in 125I seed implant brachytherapy for parotid gland cancers.

Purpose: Variability in volume delineation is a possible error source in brachytherapy. This study assessed the interobserver variations in clinical target volume (CTV) delineation in postoperative adjuvant 125I seed implant brachytherapy after parotid gland cancer surgical resection and evaluated the image fusion technique for target volume delineation. Material and Methods: Five radiation oncologists delineated gross tumor volume (GTV) and CTV in 20 patients using conventional delineation and image fusion methods. The consistency in target volume delineation was determined on the basis of differences between the oncologists. Variability was determined using Kendall's W-test, the mean conformity index (CI), the mean distance to conformity (MDC), and the center of gravity distance (CGD). Results: There were significant variations in the delineated target volumes among radiation oncologists, but the CTV consistency was significantly enhanced using the image fusion technique, based on Kendall's W, mean CI, average MDC, and average CGD, which were 0.752, 0.41, 2.75, and 4.997, respectively, using the conventional method, and 0.987, 0.86, 0.55, and 1.27, respectively, using the image fusion method. Conclusions: The interobserver variation in the delineation of the postoperative parotid target volume is large, but it can be considerably decreased using image fusion technology, which resulted in a noticeable improvement in the delineation precision of the target volume for parotid gland cancer. Thus, this technology can enhance the efficacy of 125I seed implant brachytherapy and decrease any adverse effects induced by errors in target delineation.

Acupuncture-Related Therapies for Parkinson's Disease: A Meta-Analysis and Qualitative Review.

Objective: This systematic review and meta-analysis aimed to assess the effects of the combination of acupuncture-related therapies with conventional medication compared with conventional medication in patients with Parkinson's disease (PD). Methods: A literature search within eight databases [including Medline, Embase, the Cochrane Library, PubMed, China National Knowledge Infrastructure (CNKI), China Biology Medicine (CBM), VIP, and Wanfang Database] was performed covering a time frame from their inception to August 2020. Randomized controlled trials (RCTs) comparing acupuncture-related therapies combined with conventional medication vs. conventional medication in patients with PD were eligible. Two authors independently assessed the risk of bias. Assessments were performed with the total and subscales scores of the Unified Parkinson's Disease Rating Scale (UPDRS), 39-item Parkinson's Disease Questionnaire (PDQ-39), the dosage of Madopar, Mini-Mental State Examination (MMSE), and 17-item Hamilton Depression Scale (HAMD). Data were analyzed by adopting the Cochrane Collaboration's RevMan 5.4 (Review Man, Copenhagen, Denmark); and mean effect sizes and 95% confidence intervals were estimated. Tests for heterogeneity were used to assess differences in treatment effects across different types of acupuncture used. Results: Sixty-six trials met the inclusion criteria, of which 61 trials provided data for the meta-analysis. We defined high-quality articles as those with a low risk of bias in four or more domains; and only 10 (15.15%) articles were of high quality. Compared with the controls, acupuncture-related therapies with conventional medication achieved a benefit in the primary outcomes of UPDRS (motor subscore: -3.90, -4.33 to -3.49, P < 0.01; total score: -7.37 points, -8.91 to -5.82, P < 0.001; activities of daily living subscore: -3.96, -4.96 to -2.95, P < 0.01). For the subgroup difference test among the effects of different acupuncture methods, significant differences existed in outcomes with the UPDRS-III, UPDRS-I, UPDRS-IV, and PDQ-39 scores and Madopar dosage, while non-significant differences existed with the UPDRS-total, UPDRS-II, HAMD, and MMSE scores. Conclusions: Acupuncture-related therapies combined with conventional medication may benefit individuals with PD. Our review findings should be considered with caution because of the methodological weaknesses in the included trials. Future, large randomized trials of acupuncture-related therapies for PD with high methodological quality are warranted. Systematic Review Registration: Identifier CRD42021228110.

Multifunctional TiO2/SBA-15 mesoporous silica hybrids loaded with organic sunscreens for skin application: The role in photoprotection and pollutant adsorption with reduced sunscreen permeation.

TiO2 acts as an inorganic sunscreen and photocatalyst to protect humans from environmental pollutants. We incorporated TiO2 into mesoporous silica (SBA-15) for skin application to prevent environmental stresses including UVA irradiation and pollutant invasion. Organic ultraviolet (UV)A filters such as avobenzone and oxybenzone were then loaded into mesoporous support for synergistic sunscreen efficiency. The as-prepared formulations with different TiO2 amounts (10 %-50 %) were fabricated. The pore size decreased from 4.72 to 4.00 nm following the increase in TiO2 percentage. TiO2/SBA-15 captured about 60 % fluoranthene and 80 % furfural within 3 h with no significant difference due to different TiO2 content. The in vitro photoprotection assessed by UVA/UVB ratio exhibited the increase in Boots star rating from 2 to 3 to 5 by entrapment of avobenzone into TiO2/SBA-15. Thirty-percent TiO2/SBA-15 in hydrogel decreased avobenzone and oxybenzone deposition by 70 % and 80 % compared to free form, respectively. Avobenzone and TiO2 supplementation to SBA-15 significantly alleviated skin cell death and neutrophil recruitment in the photoaged mouse skin compared to the SBA-15 application alone. Compared to the UVA-irradiated skin, 30 % TiO2/SBA-15 showed a 2.5- and 3.1-fold decline in IL-1ß and IL-6 levels, respectively. The TiO2/SBA-15 hybrid was considered non-irritant based on results of cytotoxicity assay, skin histology, and cutaneous barrier function. Our data indicate that the versatile mesoporous silica is an effective system for topical use in sunscreen and skin protection.

Aidi Injection, Compound Kushen Injection, or Kanglaite Injection: Which Is the Best Partner with Systemic Chemotherapy for Patients with HCC? A Network Meta-Analysis.

OBJECTIVE: The aim of this network meta-analysis (NMA) was to explore the effectiveness of different traditional Chinese medicine injections (TCMIs) combined with systemic chemotherapy for the treatment of hepatocellular carcinoma (HCC). METHODS: A comprehensive search for randomized controlled trials (RCTs) was performed with regard to different TCMIs for treating HCC in seven electronic databases up to November 2019. The quality assessment of the included RCTs was conducted according to the Cochrane risk of bias tool. The objective response rate (ORR), clinical benefit rate (CBR), and Karnofsky performance score (KPS) data were extracted. The network meta-analysis used the network package in Stata software to analyse the data and draw a map of the evidence summarizing the direct and indirect comparisons. RESULTS: A total of 1697 articles were retrieved through the comprehensive search. Twenty RCTs focusing on Aidi injection, compound Kushen injection, and Kanglaite injection as adjuvant therapies to chemotherapy were included, involving a total of 1418 patients. The NMA statistics showed that all three indicators (ORR, CBR, and KPS) were better in the combined treatment group of TCMIs with chemotherapy than that in the single treatment group of chemotherapy alone. Kanglaite injection tended to be better than the other two in terms of primary outcome, but there was not a significant difference. The combined treatment group had fewer adverse reactions than the single treatment group. Moreover, several articles reported that TCMIs combined with chemotherapy could increase the number of CD3+ and CD4+ T lymphocytes and the ratio of CD4+/CD8+ T lymphocytes. CONCLUSIONS: TCMIs combined with systemic chemotherapy could be an effective and safe treatment option for patients with HCC. Kanglaite injection showed a tendency to be better than the other two kinds of injections in terms of ORR. Nevertheless, additional results from multicentre trials and high-quality studies will be pivotal for supporting our findings.

Hydrogen Sulfide Prevents Elastin Loss and Attenuates Calcification Induced by High Glucose in Smooth Muscle Cells through Suppression of Stat3/Cathepsin S Signaling Pathway.

Vascular calcification can be enhanced by hyperglycemia. Elastin loss in tunica media promotes the osteogenic transformation of smooth muscle cells (SMCs) and involves arterial medial calcification (AMC) that is associated with a high incidence of cardiovascular risk in patients with type 2 diabetes. Here, we tested whether hydrogen sulfide (H2S), an endogenous gaseous mediator, can prevent elastin loss and attenuate calcification induced by high glucose in SMCs. Calcification was induced by high glucose (4500 mg/L) in human aortic SMCs (HASMCs) under the condition of calcifying medium containing 10 mM ß-glycerophosphate (ß-GP). The experiments showed that NaHS (an H2S donor, 100 µM) mitigated the calcification of HASMCs treated with high glucose by decreasing calcium and phosphorus levels, calcium deposition and ALP activity and inhibited osteogenic transformation by increasing SMα-actin and SM22α, two phenotypic markers of smooth muscle cells, and decreasing core binding factor α-1 (Cbfα-1), a key factor in bone formation, protein expressions in HASMCs. Moreover, NaHS administration inhibited the activation of Stat3, cathepsin S (CAS) activity and its expression, but increased the level of elastin protein. Pharmacological inhibition or gene silencing Stat3 not only reversed elastin loss, but also attenuated CAS expression. Inhibition of CAS alleviated, while CAS overexpression exacerbated, elastin loss. Interestingly, overexpression of wild type (WT)-Stat3, but not its mutant C259S, elevated CAS protein expression and reduced elastin level. Moreover, NaHS induced S-sulfhydration in WT, but not in the C259S Stat3. These data suggest that H2S may directly regulate Cys259 residue in Stat3 and then impair its signaling function. Our data indicate that H2S may attenuate vascular calcification by upregulating elastin level through the inhibition of Stat3/CAS signaling.

Hydrogen Sulfide: Recent Progression and Perspectives for the Treatment of Diabetic Nephropathy.

Diabetic kidney disease develops in approximately 40% of diabetic patients and is a major cause of chronic kidney diseases (CKD) and end stage kidney disease (ESKD) worldwide. Hydrogen sulfide (H2S), the third gasotransmitter after nitric oxide (NO) and carbon monoxide (CO), is synthesized in nearly all organs, including the kidney. Though studies on H2S regulation of renal physiology and pathophysiology are still in its infancy, emerging evidence shows that H2S production by renal cells is reduced under disease states and H2S donors ameliorate kidney injury. Specifically, aberrant H2S level is implicated in various renal pathological conditions including diabetic nephropathy. This review presents the roles of H2S in diabetic renal disease and the underlying mechanisms for the protective effects of H2S against diabetic renal damage. H2S may serve as fundamental strategies to treat diabetic kidney disease. These H2S treatment modalities include precursors for H2S synthesis, H2S donors, and natural plant-derived compounds. Despite accumulating evidence from experimental studies suggests the potential role of the H2S signaling pathway in the treatment of diabetic nephropathy, these results need further clinical translation. Expanding understanding of H2S in the kidney may be vital to translate H2S to be a novel therapy for diabetic renal disease.

The role of glutamate transporter-1 in the acquisition of brain ischaemic tolerance in rats induced by electro-acupuncture pre-treatment.

OBJECTIVE: To investigate whether electro-acupuncture can serve as a method of inducing brain ischaemic tolerance (BIT) by encouraging the expression of glutamate transporter-1 (GLT-1) and suppressing the release of glutamate (Glu). METHODS: Sprague-Dawley (SD) rats were divided into sham, ischaemia and EA groups. EA was performed on dazhui and baihui acupoints and the rat cerebral ischaemia model was achieved by occluding the middle cerebral artery (MCA) for 2 hours, followed by reperfusion. Dialysate was collected from the striatum in vivo to detect the concentration of Glu and the expression of Glutamate Transporter-1 (GLT-1) was examined. The changes of neurological deficit scores were evaluated at 24 hours after reperfusion, while the infarct volumes of brains were then measured with 2,3,5-triphenyltetrazolium chloride (TTC) staining. RESULTS: Compared with the ischaemia group, the concentration of Glu decreased and the expression of GLT-1 increased at most of the detective time points in the EA group; the neurological deficit scores were lower and the infarct volumes were smaller in the EA group. CONCLUSION: EA can up-regulate the expression of GLT-1 and inhibit the excessive release of Glu in the striatum in the process of subsequent ischaemic-reperfusion brain injury, which may be one of the mechanisms of inducing BIT and, thus, be neuroprotective for early ischaemic brain injury.

Brain regions involved in moxibustion-induced analgesia in irritable bowel syndrome with diarrhea: a functional magnetic resonance imaging study.

BACKGROUND: Moxibustion is one of the most commonly used therapies in acupuncture practice, and is demonstrated to be beneficial for patients with diarrhea from irritable bowel syndrome (D-IBS). But its mechanism remains unclear. Because visceral hypersensitivity in IBS patients has been documented by evaluation of perceived stimulations through functional magnetic resonance imaging (fMRI) studies, we focused on observing brain imaging changes in D-IBS patients during rectal balloon distention before and after moxibustion in order to reveal its possible central mechanism and further evaluate its effect. METHODS: This clinical trial is registered under the number: ChiCTR-TRC-10000887. Eighty D-IBS patients were randomly divided into a moxibustion and sham moxibustion group (control group) for a 4-week treatment. Fifteen patients in moxibustion group and thirteen patients in control group completed two fMRI scans during a 50 and 100 ml rectal balloon distention before and after treatment. Rectal pain were obtained with a scan test. Birmingham IBS Symptom Scale and IBS Quality of Life (QOL) Scale were used to evaluate therapeutic effect. RESULTS: After treatment, the decrease in Birmingham IBS Symptom Scale and IBS QOL Scale scores in moxibustion group was significantly greater than that of control group (P < 0.01). The defecation urge threshold and the pain perception threshold of moxibustion group was also significantly higher after treatment than that of control group (P < 0.01). The decrease in pain score during the 100 ml rectal balloon distention in moxibustion group was significantly greater than that of control group (P < 0.05). There was no definite activated center during the 50 ml rectal distention in either group before treatment. After treatment, the prefrontal cortex (PFC) was affected in moxibustion group, while the PFC and the anterior cingulated cortex (ACC) were affected in control group. During the 100 ml distention before treatment in both groups, the PFC and ACC were activated. After treatment, they disappeared in moxibustion group but remained in control group. CONCLUSIONS: Moxibustion can improve symptoms and quality of life in D-IBS patients. It can also decrease rectal sensitivity. The activation of PFC and ACC during a 100 ml rectal distention disappeared after moxibustion treatment.

Anti-inflammatory diterpenoids from the root bark of Acanthopanax gracilistylus.

Five new ent-pimarane (1-3, 7, and 8) and three new ent-kaurane diterpenoids (4-6) and a new oleanane triterpene acid (9), together with 22 known compounds, were isolated from the root bark of the medicinal herb Acanthopanax gracilistylus. The structures of 1-9 were established based on the interpretation of high-resolution MS and 1D- and 2D-NMR data. The absolute configurations of 7 and 11 were determined by single-crystal X-ray diffraction and electronic circular dichroism analysis. Compounds 7 and 8 represent rare naturally occurring structures based on the devinyl ent-pimarane skeleton. Compounds 3, 10, 14, 16, and 17 exhibited potent inhibitory effects on the release of interleukin-1ß (IL-1ß), interleukin-8 (IL-8), and tumor necrosis factor (TNF-α) in lipopolysaccharide-stimulated peripheral blood mononuclear cells.

[Analysis on diagnosis and treatment of 15 cases with severe influenza A].

OBJECTIVE: To analyze the diagnosis and treatment characteristics of patients with severe Influenza A. METHOD: A retrospective investigation on the clinical manifestation, chest radiography, electronic fiber bronchoscopy and the histology of the cast, rescue course and outcome was conducted in 15 children with severe influenza A during January to May of 2013. RESULT: Eleven cases were male, the range of age was 2 to 6 years; 5 cases were female, the range of age was 1 month to 6 years, accouting for 4.2% of hospitalized children with influenza. Three patients had an underlying chronic disease, two had nephrotic syndrome, and one had congenital heart disease. All the 15 cases were diagnosed as severe influenza A virus infection complicated with pneumonia and respiratory failure, of whom 10 cases were infected with H1N1 virus , the other 5 cases could not be identified as H1N1 virus by using H1N1 kit, but none of the 15 cases were infected with H7N9 virus. Of 15 cases, 8 had atelectasis, 4 had pneumothorax, 3 had pneumomediastinum, 4 had pleural effusion, 1 had pneumorrhagia; 12 patients required mechanical ventilation. 1 only required noninvasive mask CPAP, 2 did not require assisted ventilation, they were just given mask oxygen. Seven cases' sputum culture showed combined infection with bacteria and fungi, sputum smear examination detected: G(+) cocci in 2 cases, and G(-) bacilli in the other 2. By using electronic fiber bronchoscopy, bronchial cast was detected in 5 patiens. Histological examination of the bronchial cast revealed a fibrinous exudation containing large quantity of eosinophils, neutrophils in 1 patients, fibrinous exudation and necrotic material containing large quantity of neutrophils in 4 patients. After the bronchial casts were removed, 4 patients were improved greatly. All patients were treated with postural drainage of left and right side position, massage of electric oscillation, strengthening the sputum suction aiming to improve pulmonary ventilation function. Three patients died: 1 case was compliicated with nephrotic syndrome, another case had congenital heart disease, and 1 case hads pneumorrhagia, renal failure and multiple organ dysfunction syndrome (MODS). CONCLUSION: The mortality of severe Influenza A is higher if it is complicated with underlying chronic diseases. In children undergoing rapid and progressive respiratory distress with lung atelectasis, consolidation or emphysema on chest X-ray, plastic bronchitis should be considered. Electronic fiber bronchoscopy should be performed early Lung physicotherapeutics still are important assistant measures for improving the pulmonary ventilation function.

A rapid and convenient method for preparative separation of three indissolvable polyphenols from Euphorbia pekinensis by the flexible application of solvent extraction combined with counter-current chromatography.

A rapid and convenient method was established to preparatively isolate the three ellagic acid types of compounds, which were the main polyphenols in Euphorbia pekinensis, by flexibly applying solvent extraction combined with counter-current chromatography (CCC). The total extract (extracted using 95% ethanol) of E. pekinensis was pretreated by two simple steps before CCC isolation, following the procedure: the total extract was extracted by classical solvent extraction using petroleum ether and ethyl acetate, respectively, and then the ethyl acetate extract was suspended using 95% ethanol, after being allowed to stand overnight, the sediment was obtained. Partial sediment (100 mg) was then directly separated by CCC with a two-phase solvent system composed of chloroform-95% ethanol-water-85% formic acid (50:50:50:5, v/v/v/v). About 22 mg of 3,3'-dimethoxy ellagic acid (1), 12 mg of 3,3'-di-O-methyl-4-O-(ß-D-xylopyranosyl)ellagic acid (2), and 35 mg of ellagic acid (3) with purities of 96.0, 95.2, and 95.4% were obtained respectively in one step within 4 h. After being purified by washing with methanol, the purities of the three compounds obtained were all above 98%. The purities were determined by HPLC and their chemical structures were further identified by (1)H and (13)C NMR spectroscopy. The recoveries were calculated as 84.6, 85.7, and 89.5%, respectively. The result demonstrated that the present isolation method was rapid, economical and efficient for the preparative separation of polyphenols from E. pekinensis.

Neolignanamides, lignanamides, and other phenolic compounds from the root bark of Lycium chinense.

Seven new neolignanamides (1-7), including two pairs of cis- and trans-isomers, and a new lignanamide (8) were isolated from the EtOAc-soluble fraction of an EtOH extract of the root bark of Lycium chinense, together with 22 known phenolic compounds (9-30), four of which were obtained from the genus Lycium for the first time. Compounds 5, 6, and 7 are unusual dimers having a rare connection mode between the two cinnamic acid amide units, while compounds 6, 7, and 8 are the first naturally occurring dimers derived from two dissimilar cinnamic acid amides. The cinnamic acid amides, neolignanamides, and lignanamides possess moderate radical-scavenging activity against the DPPH (2,2-diphenyl-1-picrylhydrazyl) and superoxide radicals.

Long-term changes of diffusion tensor imaging and behavioural status after acupuncture treatment in rats with transient focal cerebral ischaemia.

BACKGROUND: The effect of acupuncture treatment in cerebral ischaemia is controversial. A study was undertaken to assess its effects in rats with transient middle cerebral artery occlusion (tMCAO) and discuss its mechanisms. METHODS: 21 Sprague-Dawley rats were divided into three groups. Group 1 underwent tMCAO and subsequently received acupuncture treatment, Group 2 underwent tMCAO without treatment and Group 3 only underwent sham operation. The evolution of diffusion tensor imaging (DTI) features in ischaemic lesions from acute to chronic periods was assessed and the correlations with behavioural tests and histopathological changes were examined. RESULTS: tMCAO rats displayed side-specific sensorimotor deficits after occlusion. Behavioural scores of rats in group 1 reduced gradually with acupuncture treatment. No significant difference in lesion size on T2-weighted imaging was found between the two tMCAO groups. Relative apparent diffusion coefficient (rADC) and relative fractional anisotropy (rFA) values in the centre and at the edge of the ischaemic lesions reduced at first and then increased to varying degrees. Only changes in the rFA value at the edge of the ischaemic lesions were significantly different between the two tMCAO groups. A more significant increase in growth-associated protein B-50 and synaptophysin protein expression was found in group 1 than in the other groups. No significant correlations were found between behavioural scores, DTI appearances and immunohistochemical results. CONCLUSIONS: The acupuncture points applied were effective, and improving neuronal regeneration may have a role in the mechanism of acupuncture treatment of post-stroke paralysis of the limbs in rats. MRI, particularly the fractional anisotropy value of DTI, is an appropriate parameter to evaluate the recovery status.

Three new acetylated benzyl-beta-resorcylate glycosides from Cassia obtusifolia.

Three new acetylated benzyl-beta-resorcylate glycosides (1-3) were isolated from seeds of Cassia obtusifolia. Their structures were determined on the basis of the spectroscopic methods and physicochemical properties as 2-benzyl-4,6-dihydroxy benzoic acid-6-O-[2,6-O-diacetyl]-D-glucopyranoside (1), 2-benzyl-4,6-dihydroxy benzoic acid-6-O-[3,6-O-diacetyl]-D-glucopyranoside (2) and 2-benzyl-4, 6-dihydroxy benzoic acid-6-O-[4,6-O-diacetyl]-D-glucopyranoside (3), respectively.

Fragmentation pathways of oxygenated tetracyclic triterpenoids and their application in the qualitative analysis of Ganoderma lucidum by multistage tandem mass spectrometry.

The fragmentation pathways of oxygenated tetracyclic triterpenoids from Ganoderma lucidum were systematically studied based on interpreting the mass spectra of 44 known triterpenoids using a combination of multistage tandem mass spectrometry (MS(n)) experiments and high-resolution mass spectrometry (HRMS) analysis. In negative ion mode, the fragmentation pathways of triterpenoid acids are rather characteristic. After the prominent loss of H(2) O or CO(2), cleavages take place on the A, B, C and D rings. Interestingly, the cleavage mode is highly dependent on the positions of the carbonyl groups and hydroxyl groups in the tetracyclic skeleton. Characteristic cleavage of ring A occurs in 7-oxo-11-H or 7-oxo-11-hydroxy derivatives; characteristic cleavage of ring B occurs in the 7-oxo-11-hydroxy derivatives; characteristic cleavage of ring C occurs in the 7-hydroxy-15-oxo derivatives; while the cleavage of ring D can be observed in the majority of the compounds investigated. The odd-electron species, which disobey the 'even-electron rule', are also observed and discussed in this paper. These phenomena provide an easy way to determine the tetracyclic skeleton and distinguish the isomers of the triterpenoids from each other. What is more, the fragmentation pathways of triterpenoid alcohols were also investigated in positive ion mode. The accurate masses of the product ions were determined using quadrupole orthogonal time-of-flight (QTOF) instruments. Finally, the fragmentation rules were applied to identify the components of G. lucidum. As a result, 73 triterpenoids including 11 new ones were identified. The triterpenoids were classified into six subclasses according to their different fragmentation behaviors. The application of tandem mass spectrometry was further explored.

Benzyl-beta-resorcylates from Cassia obtusifolia.

The seed of Cassia obtusifolia Linn have yielded three new compounds, 2-benzyl-4,6-dihydroxy benzoic acid (1), 2-benzyl-4,6-dihydroxy benzoic acid-6-O-beta-D-glucopyranoside (2) and 2-benzyl-4,6-dihydroxy benzoic acid-4-O-beta-D-glucopyranoside (3). Their structures were determined by spectroscopic methods, including (1)D- and (2)D NMR spectroscopy, HR-ESI-MS, as well as by comparison of their spectral data with those of related compounds.

[Comparison of the effect of eight kinds of volatile oil of Chinese material medica on percutaneous absorption of ibuprofen in vitro].

OBJECTIVE: To compare the permeation-enhancing effect of the volatile oil of eight kinds of Chinese material medica including Fructus Evodia, Radix Saposhnikoviae, Rhizoma Atractylodes lancea Radix Aucklandiae, Radix Curcuma wenyujin, Rhizoma et Radix Notopterygii, Lignum Aquilariae Resinatum, Herba Schizonepetae on the model drug of ibuprofen in order to provide theoretic foundation for the selection and application of natural penetration enhancer. METHODS: The permeation flux of ibuprofen was determined using the penetration experiment apparatus in vitro. The content of ibuprofen was measured by HPLC. RESULTS: All the above mentioned volatile oil showed permeation enhancing effect on ibuprofen in different degrees, and the enhancive permeation multiple was 3.46, 3.00, 2. 36, 2.32, 2.28, 2.01, 1.37 and 1.29. CONCLUSION: All eight kinds of volatile oil can enhance the percutaneous absorption of ibuprofen in vitro, among which the volatile oil of Fructus Evodia and Radix Saposhnikoviae showed significant penetration enhancing effect.