Shen Yuan extract exerts a hypnotic effect via the tryptophan/5-hydroxytryptamine/melatonin pathway in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Sleep plays a critical role in several physiologic processes, and sleep disorders increase the risk of depression, dementia, stroke, cancer, and other diseases. Stress is one of the main causes of sleep disorders. Ginseng Radix et Rhizoma and Polygalae Radix have been reported to have effects of calming the mind and intensifying intelligence in Chinese Pharmacopoeia. Traditional Chinese medicine prescriptions composed of Ginseng Radix et Rhizoma and Polygalae Radix (Shen Yuan, SY) are commonly used to treat insomnia, depression, and other psychiatric disorders in clinical practice. Unfortunately, the underlying mechanisms of the SY extract's effect on sleep are still unknown. AIM OF THE STUDY: This study aimed to investigate the hypnotic effect of the SY extract in normal mice and mice with chronic restraint stress (CRS)-induced sleep disorders and elucidate the underlying mechanisms. MATERIALS AND METHODS: The SY extract (0.5 and 1.0 g/kg) was intragastrically administered to normal mice for 1, 14, and 28 days and to CRS-treated mice for 28 days. The open field test (OFT) and pentobarbital sodium-induced sleep test (PST) were used to evaluate the hypnotic effect of the SY extract. Liquid chromatography-tandem mass spectrometry and enzyme-linked immunosorbent assay were utilized to detect the levels of neurotransmitters and hormones. Molecular changes at the mRNA and protein levels were determined using real-time quantitative polymerase chain reaction and Western blot analysis to identify the mechanisms by which SY improves sleep disorders. RESULTS: The SY extract decreased sleep latency and increased sleep duration in normal mice. Similarly, the sleep duration of mice subjected to CRS was increased by administering SY. The SY extract increased the levels of tryptophan (Trp) and 5-hydroxytryptamine (5-HT) and the expression of tryptophan hydroxylase 2 (TPH2) in the cortex of normal mice. The SY extract increased the Trp level, transcription and expression of estrogen receptor beta and TPH2 in the cortex in mice with sleep disorders by decreasing the serum corticosterone level, which promoted the synthesis of 5-HT. Additionally, the SY extract enhanced the expression of arylalkylamine N-acetyltransferase, which increased the melatonin level and upregulated the expressions of melatonin receptor-2 (MT2) and Cryptochrome 1 (Cry1) in the hypothalamus of mice with sleep disorders. CONCLUSIONS: The SY extract exerted a hypnotic effect via the Trp/5-HT/melatonin pathway, which augmented the synthesis of 5-HT and melatonin and further increased the expressions of MT2 and Cry1.

Anti-inflammatory effect and pharmacokinetics of dehydroandrographolide, an active component of Andrographis paniculata, on Poly(I:C)-induced acute lung injury.

Acute lung injury (ALI) is a common and critical respiratory disorder caused by various factors, with viral infection being the leading contributor. Dehydroandrographolide (DAP), a constituent of the Chinese herbal plant Andrographis paniculata, exhibits a range of activities including anti-inflammatory, in vitro antiviral and immune-enhancing effects. This study evaluated the anti-inflammatory effects and pharmacokinetics (PK) profile of DAP in ALI mice induced by intratracheal instillation of Poly(I:C) (PIC). The results showed that oral administration of DAP (10-40 mg/kg) effectively suppressed the increase in lung wet-dry weight ratio, total cells, total protein content, accumulation of immune cells, inflammatory cytokines and neutrophil elastase levels in bronchoalveolar lavage fluid of PIC-treated mice. DAP concentrations, determined by an LC-MS/MS method, in plasma after receiving DAP (20 mg/kg) were unchanged compared to those in normal mice. However, DAP concentrations and relative PK parameters in the lungs were significantly altered in PIC-treated mice, exhibiting a relatively higher maximum concentration, larger AUC, and longer elimination half-life than those in the lungs of normal mice. These results demonstrated that DAP could improve lung edema and inflammation in ALI mice, and suggested that lung injury might influence the PK properties of DAP, leading to increased lung distribution and residence. Our study provides evidence that DAP displays significant anti-inflammatory activity against viral lung injury and is more likely to distribute to damaged lung tissue.

Isolation and identification of 3,4-seco-labdane diterpenoids from Callicarpa nudiflora and investigation of their cytotoxicity against HepG2 cells.

Twenty-one previously undescribed compounds, including nineteen 3,4-seco-labdanes (nudiflopenes P-W, Y, AI-JI), one 3,4-seco-pimarane (nudiflopene X), and one labdane (nudiflopene Z), along with nine known compounds (one 3,4-seco-pimarane and eight 3,4-seco-labdanes) were isolated from the leaves of Callicarpa nudiflora Hook. Et Arn. The structures of these compounds were elucidated by high-resolution electrospray ionization mass spectrometry and one- and two-dimensional nuclear magnetic resonance spectroscopy. In addition, configurations of the isolated compounds were determined by electronic circular dichroism, DP4+ probability analysis, and single-crystal X-ray diffraction experiments. All undescribed compounds were evaluated for their cytotoxicity against HepG2 cells in vitro, among which compound 12 exhibited a moderate activity with an IC50 value of 27.8 µM.

Nontargeted metabolomic analysis of four different parts of Actinidia arguta by UPLC-Q-TOF-MSE.

Actinidia arguta, an edible berry plant with high nutritional values, has been widely used in Asian countries as a food and traditional medicinal herb. The well-recognized health-promoting properties of A. arguta were associated with its bioactive components in its different botanical parts. To rapidly screen and identify chemical components and simultaneously determine the potential metabolites from different parts of A. arguta, UPLC-Q-TOF-MSE coupled with UNIFI platform and multivariate statistical analysis approach was established in this study. As a result, a total of 107 components were identified from the four different parts of A. arguta, in which 31 characteristic chemical markers were discovered among them, including 12, 8, 6, and 5 compounds from the fruits, leaves, roots, and stems, respectively. These results suggested that the combination of UPLC-Q-TOF-MSE and metabolomic analysis is a powerful method to rapidly screen characteristic markers for the quality control of A. arguta.

The impact of phyto- and endo-cannabinoids on central nervous system diseases：A review.

Background and aim: Cannabis sativa L. is a medicinal plant with a long history. Phyto-cannabinoids are a class of compounds from C. sativa L. with varieties of structures. Endocannabinoids exist in the human body. This article provides an overview of natural cannabinoids (phyto-cannabinoids and endocannabinoids) with an emphasis on their pharmacology activities. Experimental procedure: The keywords "Cannabis sativa L″, "cannabinoids", and "central nervous system (CNS) diseases" were used for searching and collecting pieces of literature from PubMed, ScienceDirect, Web of Science, and Google Scholar. The data were extracted and analyzed to explore the effects of cannabinoids on CNS diseases. Result and conclusion: In this paper, schematic diagrams are used to intuitively show the phyto-cannabinoids skeletons' mutual conversion and pharmacological activities, with special emphasis on their relevant pharmacological activities on central nervous system (CNS) diseases. It was found that the endocannabinoid system and microglia play a crucial role in the treatment of CNS diseases. In the past few years, pharmacological studies focused on Δ9-THC, CBD, and the endocannabinoids system. It is expected to encourage new studies on a more deep exploration of other types of cannabinoids and the mechanism of their pharmacological activities in the future.

Kunxian Capsule Extract Inhibits Angiogenesis in Zebrafish Embryos via PI3K/AKT-MAPK-VEGF Pathway.

OBJECTIVE: To investigate the anti-angiogenic activity of Kunxian Capsule (KX) extract and explore the underlying molecular mechanism using zebrafish. METHODS: The KX extract was prepared with 5.0 g in 100 mL of 40% methanol followed by ultrasonication and freeze drying. Freeze dried KX extract of 10.00 mg was used as test stock solution. Triptolide and icariin, the key bioactive compounds of KX were analyzed using ultra-high performance liquid chromatography. The transgenic zebrafish Tg(flk1:GFP) embryos were dechorionated at 20-h post fertilization (hpf) and treated with PTK 787, and 3.5, 7, 14 and 21 µg/mL of KX extract, respectively. After 24-h post exposure (hpe), mortality and malformation (%), intersegmental vessels (ISV) formation, and mRNA expression level of angiogenic pathway genes including phosphoinositide 3-kinase (PI3K), protein kinase B (AKT), extracellular signal-regulated kinases (ERKs), mitogen-activated protein kinase (MAPK), vascular endothelial growth factor (VEGF) and fibroblast growth factor (FGF-2) were determined. Further, the embryos at 72 hpf were treated with KX extract to observe the development of sub-intestinal vein (SIV) after 24 hpe. RESULTS: The chromatographic analysis of test stock solution of KX extract showed that triptolide and icariin was found as 0.089 mg/g and 48.74 mg/g, respectively, which met the requirements of the national drug standards. In zebrafish larvae experiment, KX extract significantly inhibited the ISV (P<0.01) and SIV formation (P<0.05). Besides, the mRNA expression analysis showed that KX extract could significantly suppress the expressions of PI3K and AKT, thereby inhibiting the mRNA levels of ERKs and MAPK. Moreover, the downstream signaling cascade affected the expression of VEGF and its receptors (VEGFR and VEGFR-2). FGF-2, a strong angiogenic factor, was also down-regulated by KX treatment in zebrafish larvae. CONCLUSION: KX extract exhibited anti-angiogenic effects in zebrafish embryos by regulating PI3K/AKT-MAPK-VEGF pathway and showed promising potential for RA treatment.

Exploring Active Ingredients, Beneficial Effects, and Potential Mechanism of Allium tenuissimum L. Flower for Treating T2DM Mice Based on Network Pharmacology and Gut Microbiota.

Forty compounds were isolated and characterized from A. tenuissimum flower. Among them, twelve flavonoids showed higher α-glucosidase inhibition activities in vitro than acarbose, especially kaempferol. The molecular docking results showed that the binding of kaempferol to α-glucosidase (GAA) could reduce the hydrolysis of substrates by GAA and reduce the glucose produced by hydrolysis, thus exhibiting α-glucosidase inhibition activities. The in vivo experiment results showed that flavonoids-rich A. tenuissimum flower could decrease blood glucose and reduce lipid accumulation. The protein expression levels of RAC-alpha serine/threonine-protein kinase (AKT1), peroxisome proliferator activated receptor gamma (PPARG), and prostaglandin G/H synthase 2 (PTGS2) in liver tissue were increased. In addition, the Firmicutes/Bacteroidetes (F/B) ratio was increased, the level of gut probiotics Bifidobacterium was increased, and the levels of Enterobacteriaceae and Staphylococcus were decreased. The carbohydrate metabolism, lipid metabolism, and other pathways related to type 2 diabetes mellitus were activated. This study indicating flavonoids-rich A. tenuissimum flower could improve glycolipid metabolic disorders and inflammation in diabetic mice by modulating the protein expression and gut microbiota.

Glycosides and flavonoids from the extract of Pueraria thomsonii Benth leaf alleviate type 2 diabetes in high-fat diet plus streptozotocin-induced mice by modulating the gut microbiota.

Twenty glycoside derivatives and nine flavonoids from the leaves of Pueraria (P. thomsonii) were isolated by column chromatography and characterized by nuclear magnetic resonance spectroscopy (NMR) and high performance liquid chromatography (HPLC). The contents of twenty glycosides and nine flavonoids from the extract of P. thomsonii leaf (PL) were 173.3 mg g-1 and 134.7 mg g-1, respectively. Two flavonoids with the highest content were robinin (49.28 mg g-1) and puerarin (42.87 mg g-1). Six flavonoids, i.e. puerarin, robinin, rutin, quercetin, quercitrin, and kaempferol showed more inhibitory effects against α-glucosidase than acarbose. PL could effectively increase the level of insulin, decrease the content of fasting blood glucose, reduce lipid accumulation in plasma, ameliorate oxidative injury and inflammation, and relieve liver and kidney damage in diabetic mice. Moreover, PL could increase intestinal probiotics to improve metabolic disorders caused by diabetes and decrease the level of Clostridium celatum to relieve inflammation. This study suggested that PL or its glycoside derivatives and flavonoids regulating glycolipid metabolism and inflammation levels might have the potential to be used to control type 2 diabetes.

Diosmetin has therapeutic efficacy in colitis regulating gut microbiota, inflammation, and oxidative stress via the circ-Sirt1/Sirt1 axis.

Diosmetin (3',5,7 -trihydroxy-4'-methoxy flavone) is a natural flavonoid compound in the citrus species, it exhibits a variety of pharmacological activities, but little is known of its effects on colitis. In this study we evaluated the therapeutic effects of diosmetin on mouse models of chronic and acute colitis. Chronic colitis was induced in mice by drinking water containing 3% dextran sulfate sodium (DSS) from D0 to D8, followed by administration of diosmetin (25, 50 mg · kg-1 · d-1) for another 8 days. Acute colitis was induced by drinking water containing 5% DSS from D0 to D7, the mice concomitantly received diosmetin (25, 50 mg · kg-1 · d-1) from D1 to D7. During the experiments, body weight and disease activity index (DAI) were assessed daily. After the mice were sacrificed, colon tissue and feces samples were collected, and colon length was measured. We showed that in both models, diosmetin administration significantly decreased DAI score and ameliorated microscopic colon tissue damage; increased the expression of tight junction proteins (occludin, claudin-1, and zonula occludens-1), and reduced the secretion of proinflammatory cytokines IL-1ß, IL-6, TNF-α, and Cox-2 in colon tissue. We found that diosmetin administration remarkably inhibited colon oxidative damage by adjusting the levels of intracellular and mitochondrial reactive oxygen species, GSH-Px, SOD, MDA and GSH in colon tissue. The protection of diosmetin against intestinal epithelial barrier damage and oxidative stress were also observed in LPS-treated Caco-2 and IEC-6 cells in vitro. Furthermore, we demonstrated that diosmetin markedly increased the expression of Nrf2 and HO-1 and reduced the ratio of acetylated NF-κB and NF-κB by activating the circ-Sirt1/Sirt1 axis, which inhibited oxidative stress and inflammation in vivo and in vitro. Diosmetin reversed the effects of si-circSirt1 and si-Sirt1 in LPS-treated Caco-2 and IEC-6 cells. When the gut microbiota was analyzed in the mouse model of colitis, we found that diosmetin administration modulated the abundance of Bacteroidetes, Actinobacteria, Cyanobacteria and Firmicutes, which were crucial for inflammatory bowel disease. Our results have linked colitis to the circ-Sirt1/Sirt1 signaling pathway, which is activated by diosmetin. The results imply that diosmetin may be a novel candidate to alleviate DSS-induced colitis and can be a lead compound for future optimization and modification.

Clinical Response to Traditional Chinese Herbs Containing Realgar (As2S2) is Related to DNA Methylation Patterns in Bone Marrow DNA from Patients with Myelodysplastic Syndrome with Multilineage Dysplasia.

PURPOSE: DNA methylation is known to play an important role in myelodysplastic syndrome (MDS). We previously showed that Chinese herbs (CHs) containing realgar (As2S2) were effective at treating MDS with multilineage dysplasia (MDS-MLD). We tested whether the response to CH treatment was related to changes in DNA methylation in MDS-MLD. PATIENTS AND METHODS: First, the Illumina methylation 850K array BeadChip assay was used to assess the pretreatment methylation status in bone marrow cells from eight MDS-MLD patients and 3 healthy donors. The eight MDS-MLD patients were then treated with CHs for six months, the arsenic concentration was measured following treatment. The patients were subsequently divided into "effective" and "ineffective" treatment response groups and the DNA methylation patterns of the two groups were compared. Finally, the BeadChip data were validated by pyrosequencing. RESULTS: Five of the eight MDS-MLD patients showed hematological improvement (effective-treatment group), while three showed disease progression (ineffective-treatment group) (positive response rate: 62.5%). The arsenic concentrations in the patients ranged from 26.60 to 64.16 µg/L (median 48.4 µg/L) and were not significantly different between the two groups (p = 0.27). Compared with the healthy controls, three genes were hypomethylated and 110 were hypermethylated in the ineffective-treatment group. However, in the group showing hematological improvement, 102 genes were markedly hypomethylated and 87 hypermethylated. The effective-treatment group had a higher proportion of hypomethylated sites than the ineffective-treatment group (53.9% vs 2.6%, respectively; chi-square test) (p < 0.0001). Two hypermethylated and two hypomethylated genes were selected for validation by pyrosequencing (all p < 0.05). CONCLUSION: MDS-MLD patients may present different DNA methylation subtypes. CHs containing realgar may be effective for treating MDS-MLD patients with the hypomethylation subtype.

[Analysis on the rule of clinical acupoint selection in treatment of puerperal insufficient lactation with acupuncture and moxibustion].

The rule and characteristics of clinical acupoint selection were explored in treatment of puerperal insufficient lactation with acupuncture and moxibustion. The clinical articles on the treatment of puerperal insufficient lactation with acupuncture and moxibustion were retrieved from the databases of CNKI, VIP and Wanfang from the date of establishment to June 1, 2019. The articles were screened in accordance with the inclusion and exclusion criteria. The prescriptions of acupuncture and moxibustion were extracted and normalized. Using Microsoft Excel 2016 software, the use frequency, meridians involved and acupoint distributions were analyzed. Using SPSS Statistics 21.0 software, the cluster and factor analyses were conducted. Totally, 102 articles were included,108 acupoint prescriptions were extracted, 65 acupoitns were designed and the total use frequency was 654 times. The top three acupoints used in treatment of puerperal insufficient lactation were Danzhong (CV 17), Rugen (ST 18) and Shaoze (SI 1). The most frequently involved meridians were the stomach meridian, the conception vessel, the small intestine meridian and the liver meridian. The acupoints were mainly distributed in the chest and four extremities. It was showed in cluster analysis that Rugen (ST 18), Shaoze (SI 1), Zusanli (ST 36) and Danzhong (CV 17) represent 3 clusters and a total of 7 common factors were extracted. The acupoint selection is based on syndrome differentiation in treatment of puerperal insufficient lactation with acupuncture and moxibustion, of which, the local acupoints are predominated and the distal acupoints are combined.

[Changes in chemical compositions of Chrysanthemi Flos after frying and protective effects on CCl_4-induced acute liver injury in mice].

To reveal the processing mechanism of Chrysanthemi Flos from the changes of chemical compositions after frying and its effect on the efficacy of liver protection. Ultra performance liquid chromatography-quadrupole-time of flight-mass spectrometry(UPLC-Q-TOF-MS) and ultra high performance liquid chromatography(HPLC) were used for the qualitative and quantitative researches of chemical compositions before and after Chrysanthemi Flos frying. Progenesis QI and SPSS software were used for principal component analysis(PCA), partial least squares discriminant analysis(PLS-DA), variable importance projection(VIP) analysis and t-test to identify the compositions with significant changes. Pharmacodynamics experiment was used to investigate the protective effect of crude and fried Chrysanthemi Flos on CCl_4-induced acute liver injury in mice. According to mass spectrometry data, there were 28 chemical compositions in crude and fried Chrysanthemi Flos, mainly including flavonoids and organic acids. 13 compositions such as luteolin, apigenin and luteolin glycoside were increased significantly after frying, while 7 compositions such as chlorogenic acid, luteolin-7-O-glucuronide and apigenin-7-O-glucuronide were decreased significantly after frying. Through principal component analysis, crude and fried Chrysanthemi Flos products were divided into two categories, indicating that there were internal differences in quality. The results of liver injury protection experiment in mice showed that the AST, ALT and MDA contents were significantly decreased and SOD level was increased in mice with liver injury in both the high and medium dose groups. Histopathological examination showed that crude and fried Chrysanthemi Flos can protect the liver by reducing inflammatory cell infiltration, reducing steatosis, and repairing damaged liver cells. The results of this study showed that the chemical compositions had obvious changes after frying, and both crude and fried Chrysanthemis Flos had protective effects on CCl_4-induced acute liver injury in mice. In addition, in the range of high, medium and low doses, the liver protection effect of crude and fried Chrysanthemi Flos increased with the increase of dose. The experiment results provided reference for the mechanism of fried Chrysanthemi Flos and clinical selection of processed products.

Discovery and identification of antithrombotic chemical markers in Gardenia Fructus by herbal metabolomics and zebrafish model.

ETHNOPHARMACOLOGICAL RELEVANCE: Gardenia Fructus (GF), a traditional Chinese medicine for clearing heat and purging fire, has been reported to use to treat thrombotic related diseases, but the antithrombotic components are not clear. AIM OF THE STUDY: To develop efficient research methods for discovering some representative antithrombotic compounds of GF. MATERIALS AND METHODS: AB line zebrafish induced by arachidonic acid (AA) was used as a fast and trace-sample-required valuation model for antithrombptic effect of GF samples. Among nine samples of GF from different production areas, two samples with the largest difference in bioactivity were selected for downstream analysis. High-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF/MS) was applied to detect compounds in the GF samples. And herbal metabolomics and grey correlation analysis (GCA) were used to identify crucial compounds with potential antithrombotic activity. Then the bioactivity of those important compounds was verified on the zebrafish model. Network pharmacology was used to explore the protein targets and signaling pathways of these compounds. RESULTS: Among the GF samples, S1 (Huoshan City, Anhui Province), and S6 (Jichun City, Hubei Province), significantly differed in thrombus inhibiting bioactivity. HPLC-Q-TOF/MS identified a total of 614 compounds in each GF sample. 19 compounds were selected as important potential variables from metabolomics data by orthogonal partial least squares discriminant analysis (OPLS-DA). And 10 compounds among them were further found to be positively correlated with the antithrombotic bioactivity of GF by GCA. Finally, 3 compounds in them, geniposide, citric acid, and quinic acid, were confirmed as representative antithrombotic chemical markers of GF. Using network pharmacology analysis, some key protein targets, such as proto-oncogene tyrosine-protein kinase Src (SRC) and cyclin-dependent kinase 2 (CDK2), and some signaling pathways were found to supply powerful evidence about antithrombotic mechanisms of three compounds and GF. CONCLUSIONS: This research have succeeded to discover and identify three representative antithrombotic compounds of GF using an efficient integrated research strategy we established, an Omics Discriminant-Grey Correlation-Biological Activity strategy. The antithrombotic chemical makers we found could also contribute to provided more accurate index components for comprehensive quality control of GF.

An effective method for the semi-preparative isolation of high-purity anthocyanin monomers from grape pomace.

Grape (Vitis vinifera L.) pomace, the residue of red wine production, is a good source material for production of anthocyanins. In this study, an effective and simple method for semi-preparative isolation of anthocyanins from grape pomace was developed. Ultrasonication with acidified MeOH was used to extract anthocyanins, with 56.15 mg total anthocyanins being obtained from 50 g grape pomace. Crude extracts were purified by XAD-7HP column chromatography, followed by isolation of the anthocyanin mixtures using semi-preparative HPLC, and subsequent identification of anthocyanin monomers by HPLC-DAD-MS/MS. Fourteen anthocyanins were isolated with high purities (≥90%), among which were non-acylated and acylated anthocyanins, and their cis and trans isomers. It is believed this is the first time that nearly all primary anthocyanin monomers in grapes have been isolated simultaneously using a single-step semi-HPLC procedure. The findings of this study will contribute to further research on anthocyanin monomers and profitable utilization of grape pomace.

[Clinical observation of acupuncture at Qianlianquan (Extra) and eight confluent points on chronic pharyngitis].

OBJECTIVE: To observe the clinical therapeutic effect of acupuncture at Qianlianquan (Extra) and eight confluent points on chronic pharyngitis. METHODS: A total of 42 patients with chronic pharyngitis were randomized into an observation group and a control group, 21 cases in each one. In the observation group, acupuncture was applied at Qianlianquan (Extra) and eight confluent points [Neiguan (PC 6), Waiguan (TE 5), Lieque (LU 7), Houxi (SI 3), Gongsun (SP 4), Shenmai (BL 62), Zhaohai (KI 6) and Zulinqi (GB 41)], once a day, 6 days as one course, with an interval of 1 day between the courses, 4-course treatment was required. In the control group, Qinghou Liyan granules were taken orally 5 g each time, 3 times a day, 7 days as one course and 4 courses were required. Before and after treatment, symptoms and signs scores of chronic pharyngitis and the contents of IL-2 and TNF-a in serum were compared in the two groups. RESULTS: Compared before treatment, symptoms and signs scores of chronic pharyngitis and the contents of IL-2 and TNF-a in serum were improved in both groups (all P<0.05), and the changes in the observation group were superior to the control group (all P<0.05). The total effective rate was 90.5% (19/21) in the observation group, which was superior to 81.0% (17/21) in the control group (P<0.05). CONCLUSION: The therapeutic effect of acupuncture at Qianlianquan (Extra) and eight confluent points on chronic pharyngitis is superior to conventional Chinese patent medication treatment by Qing Liyan granules.

[Study on liver protection and hepatotoxicity of saikosaponin a based on zebrafish model].

Bupleuri Radix has both liver protection and hepatotoxicity. Saponins are the main pharmacodynamic and toxic components of Bupleuri Radix. Based on zebrafish physical model and the model of alcoholic fatty liver( AFL) pathology,the liver toxic and protective effect of saikosaponin a( SSa) were assessed. The results indicated that 1. 77 µmol·L-1 SSa showed protective effect to AFL zebrafish. 5. 30 µmol·L-1 SSa was hepatotoxic to healthy zebrafish,but it showed protective effect to AFL zebrafish. 5. 62 µmol·L-1 SSa was hepatotoxic to healthy and AFL zebrafish. This study is benefit for clinical safety of saikosaponin a.

[Clinical observation on the treatment of hand dysfunction after stroke with chicken-claw needling at Shangbaxie (Extra) and Hegu (LI 4) combined with acupuncture at Houxi (SI 3)].

OBJECTIVE: To observe the difference in clinical efficacy between chicken-claw needling at Shangbaxie (Extra) and Hegu (LI 4) combined with acupuncture at Houxi (SI 3) and the conventional acupuncture at the 3 points for the treatment of hand dysfunction after stroke. METHODS: Forty-two patients were divided into an observation group and a control group according to the random number table, 21 cases in each one. The chicken-claw needling was used at Shangbaxie (Extra) and Hegu (LI 4) on the affected side combined with acupuncture at Houxi (SI 3) in the observation group. The conventional acupuncture was performed at the same point as the observation group in the control group, once a day, 6 days for a course, 1 day of interval after a course, and the therapeutic effect was observed after 5 courses. The simplifying Fugl-Meyer Motor Function Rating Scale (FMA), the modified Barthel index and the Brunnstrom grading criteria were used to evaluate the hand function of the two groups before and after treatment. RESULTS: The FMA score, Barthel index and Brunnstrom grade were improved after treatment in the two groups (all P<0.05), and the FMA score, Barthel index and Brunnstrom grade in the observation group were better than those in the control group (all P<0.05). CONCLUSION: Chicken-claw needling at Shangbaxie (Extra) and Hegu (LI 4) combined with acupuncture at Houxi (SI 3) can effectively treat hand dysfunction after stroke, and the curative effect is better than the conventional acupuncture at the 3 points.

Geographical origin of Chinese wolfberry (goji) determined by carbon isotope analysis of specific volatile compounds.

Chinese wolfberry or goji berry (Lycium barbarum) is an important traditional Chinese medicine. Its price and function has a close correlation with its geographical provenance. Illegal mislabeling motivated by commercial gains brings serious food safety problems and damages consumer confidence. In this work, a novel analytical strategy combined with chemometrics statistic tools was developed to determine the geographical origin of wolfberries from different provinces in China. Stable carbon isotopic ratios (δ13C) of wolfberry volatile compounds (i.e. limonene, tetramethylpyrazine, safranal, geranylacetone, and ß-ionone) were determined by gas chromatography-combustion-isotope ratio mass spectrometry (GC-IRMS) with headspace-solid phase micro extraction (HS-SPME). Five types of SPME fiber (i.e. DVB/CAR/PDMS, CAR/PDMS, PDMS/DVB + OC, PDMS, and PA), extraction time, temperature and GC-IRMS conditions were comprehensively optimized to obtain the best adsorption of volatile compounds in wolfberry. Method integrity was assessed by comparing volatiles extracted using HS-SPME GC-IRMS with direct injection GC-IRMS and were in good agreement with each other. The geographical variations of volatile compounds using one-way analysis of variance (ANOVA) were explored for individual δ13C values in wolfberry samples from Gansu, Ningxia and Qinghai. Geographical origin of wolfberry was differentiated by linear discrimination analysis (LDA), with an accuracy of 89.16%, 87.77% and 85.87% for these three provinces, respectively. These results showed the combination of SPME and IRMS provides a rapid and valid method to determine the geographical origin of wolfberry.

[Establishment of DNA identity card and analysis of genetic similarity among 58 varieties in Lonicera japonica].

A total of 58 varieties in Lonicera japonica from 20 producing areas were amplified by 22 pairs of SSR primers. Seven pairs of polymorphic primers were screened and their primers were used to establish DNA identity card and analyze genetic similarity.All the 58 varieties could be distinguished each other by the DNA identity card constituted by 7 pairs of core SSR primers.The genetic similarity coefficients of 58 varieties ranged from 0.366 7 to 0.916 7 by using PopGene32（vesion1.32）. Furthermore, all the varieties consistency were classified into 4 groups and constructed an evaluation table according to cluster analysis by an un-weighted pair-group average method with arithmetic mean. As expected, the results of cluster and evaluation table reflected 58 varieties relatives, which provide reference information for the selection of fine germplasm of L. japonica and the theoretical basis for the study of Dao-di herbs.

[Comparison of three different thermophilic beta-glucosidases for baicalin transformation].

Three different beta-glycosidase sequences of Ttebgl3, Tpebgl1 and Tpengl3 from Thermotoga thermarum DSM 5069 and Thermotoga petrophila RKU-1 were analyzed. Also, the influence of temperature, pH, concentration of DMSO, metal ions and kinetic constant on catalytic conversion of baicalin had been compared. The results indicated that the optimal pH and optimum temperature for transformation of baicalin was 4.5 85 °C, 5.0 80 °C and 5.5 80 °C, respectively. The family GH3 beta-glycosidase Ttebgl3 and Tpebgl3 had the better DMSO tolerance. The activation effect of the metal ions on the catalytic conversion of baicalin was not obvious, and the inhibition of the GH3 family beta glucosidase was significantly stronger than that of the GH1 family. The kinetic constants of three different beta-glucosidases catalyzed baicalin were significantly different. The Km and Vmax values of Tpebgl1, Tpebgl3 and Ttebgl3 were 0.029 2 mmol·L⁻¹ 4.85 U·mg⁻¹, 0.268 6 mmol·L⁻¹ 121.04 U·mg⁻¹ and 0.391 8 mmol·L⁻¹ 308.90 U·mg⁻¹, respectively. Family GH3 beta-glycosidase converted more baicalin than family GH1 with the optimal conditions, 0.02 g baicalin, and the conversion rate was 68%, 97.3%, 97.31% respectively. The results of the study provided a guarantee for the transformation of baicalin.