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[Fungi isolated from diseased medicinal plants].

One hundred and forty-four fungal isolates were obtained from diseased Paeonia albiflora Pall. var. trichocarpa Bung., Astragalus membranaceus Bung., Lithospermum erythrorhizon Sieb. et Zucc., Ledebouriella seseloides Wolff and Bupleurum falcatum L. which were collected in the test field of Tsukuba Medicinal Plant Research Station, National Institute of Hygienic Sciences. Most of them were identified into 15 genera containing 8 species. Fungal species presumed to be pathogens of the host plants were as follows: Cladosporium paeoniae, Pestalotia paeoniicola, Glomerella cingulata, Hainesia lythri, Guignardia sp. and Alternaria sp. from P. albiflora, Fusarium spp., Rhizoctonia spp. and Neocosmospora vasinfecta from A. membranaceus, Colletotrichum gloeosporioides from L. erythrorhizon, Rhizoctonia sp., Fusarium spp., Phoma sp. and Pyrenochaeta sp. from L. seseloides, and Fusarium sp., Alternaria alternata, Phyllosticta sp., Phoma sp., Phomopsis sp. and C. gloeosporioides from B. falcatum. Roots of B. falcatum were found to be parasitized by Meloidogyne sp.

Astragalus propinquus↗

[A histochemical study of saikosaponins].

The root of Bupleurum chinese and B.scorzonerifolium was examined histochemically in order to clarify the distribution of saikosaponins in various tissues and parts of the root. By means of qualitative and quantitative analyses, saikosaponins, the bioactive principles in the root, were found to be abundant in the cortex outside the cambium and very little in the xylem of the root. It was also found that the thinner root hairs had the highest contents of saikosaponins, but decreased towards the thicker root head.

Anti-Inflammatory Agents, Non-Steroidal↗

[Effect of saikosaponins on epileptic seizure and EEG in pentetrazole-induced chronic kindling rats].

OBJECTIVE: To study the effect of saikosaponins, the active ingredients of Bupleurum chinense DC, on epileptic seizure and EEG of pentetrazole (PTZ)-induced chronic kindling rats. METHODS: Forty-eight healthy Sprague-Dawley rats were randomly divided into 6 equal groups, namely the blank control group, normal saline (NS) group, sodium valproate (VPA) group, and 3 saikosaponins groups of high, medium and small doses. Except those in the blank control group, the rats in the other groups were all given different treatments as specified prior to intraperitoneal PTZ injection to induce kindling on a daily basis for 4 consecutive weeks. Epileptic seizures of the rats were recorded during the treatment and EEG recorded at the end of the treatments. RESULTS: Seizure frequency in the 3 saikosaponins groups decreased 2 weeks later, which was especially obvious in the high-dose group (P<0.05). The kindling rate was significantly lower in high-dose saikosaponins group than in the other treatment groups after 4 weeks of the treatment (P<0.05), with also less intense seizure onset (P<0.01) and differences in the wave form of EEG. CONCLUSION: Saikosaponins can inhibit PTZ-induced epileptic seizure in kindling rats and antagonize the kindling effect of PTZ.

Animals↗

[Saikosaponins inhibit increased glutamate and GABA expressions in the hippocampus of pentetrazole-induced slow kindling rats].

OBJECTIVE: To study the effect of saikosaponins, the active ingredients of Bupleurum chinense DC. on glutamate and GABA expressions in the hippocampus of slow kindling rats induced by pentetrazole. METHODS: Forty-eight healthy Sprague-Dawley rats were randomly divided into 6 equal groups, namely the blank control group (group A), normal saline (NS) group (group B), sodium valproate group (group C), and 3 saikosaponins groups of high, medium and small doses (groups D, E, and F, respectively). The rats in each group other than group A were given corresponding treatments after slow kindling by pentetrazole. After 4 weeks of treatment, the rats were sacrifices and the brain tissues were sampled, sliced and stained by immunohistochemically, and the results were analyzed according to the positive cell number and gray scale. RESULTS: In CA1 region, the glutamate-positive cell number and gray scale of group B was significantly different from the other groups (P<0.05), but such difference was not observed in the CA2 and DG (P>0.05); In CA1, CA2 and DG of the hippocampus, the GABA-positive cell number of group B was significantly greater but the gray scale lower than those of the other groups (P<0.05). In CA1 and CA2 regions of the hippocampus, the glutamate- and GABA-positive cell ratio of group B was significantly lower than that of the other groups (P<0.05), but in CA1, CA2, and DG region of the hippocampus, the ratio of gray scale between glutamate- and GABA-positive cells was comparable between the groups (P>0.05). CONCLUSION: The expression of glutamate and GABA, especially the latter, increased in chronic kindling rat hippocampus. Saikosaponins intervene in such changes of glutamate and GABA to contain their expressions within normal range, which may be one of the mechanisms of saikosaponins to inhibit slow kindling induced by pentetrazole.

Animals↗

[Dynamic variation of saikosaponin contents].

In this paper both the cultivated and wild Bupleurum chinense and B. scorzonerifolium from northeast China of different growing periods and ages were analyzed for the contents of saikosaponins a and d by HPLC and total saikosaponins by spectrophotometry. The results indicated that the saikosaponin content of B. chinense was higher than that of B. scorzonerifolium, and that the saikosaponin contents of the cultivated species were higher than or almost equal to those of the wild ones. It was also found that both the cultivated and wild B. chinense and B. scorzonerifolium featured the highest contents of saikosaponins in the nutrition-growing period.

Anti-Inflammatory Agents, Non-Steroidal↗

[Preliminary study of traditional Chinese medicine treatment of minimal brain dysfunction: analysis of 100 cases].

This paper reports the clinical treatment of 100 patients with minimal brain dysfunction (MBD), their age ranged from 7-14.2 years, and the average age was 10.5. Patients were divided into TCM and WM group randomly. The TCM group (80 cases) were treated with the therapy of subdueing hyperactivity of the Liver and invigorating the function of the Spleen by using: Bupleurum chinense, Scutellaria baicalensis, Astragalus membranaceus, Codonopsis pilosula, Ligustrum lucidum, Lophatherum gracile and thread of ivory. The WM group (20 cases) were treated with retalin 5-15 mg twice daily. One course of treatment lasted one month, and effects were evaluated after 1-3 courses of treatment. The results were as follows: In the TCM group, 23 cases were cured (clinical symptom and sign disappeared, 10 IQ units were raised in their intelligence, abnormal electroencephalogram was recovered and there was no recurrence during the first six months after recovery). 46 cases were improved (clinical symptom and sign markedly improved, 4 IQ units raised in intelligence and electroencephalogram improved), and 11 cases ineffective. The effective rate was 86.25%. In the WM group, 6 cases were cured, 12 cases improved and 2 cases ineffective, the clinical effective rate being 90.0%. There was no significant difference between the two groups in this study, but, the side-effects of the TCM group was less than the WM group, and the TCM group had more beneficial effects to improve intelligence, enuresis and the black of orbits. The study showed that the Verbal Scale in Wechsler Intelligence Scale for children is one of the indications of diagnosis and evaluating of the curative effect of MBD.

Adolescent↗

[Phytotherapeutic aspects of diseases of the cardiovascular system. 5. Saponins and possibilities of their use in prevention and therapy].

The summarizing paper deals with the structure and biological effects of saponins generally and in relation with the treatment and prevention of diseases of the heart and circulatory system. In this field, mainly the saponins from the plants of the genera Panax, Gynostemma or Bupleurum are of use. Also soya saponins and saponins of the genera Astragalus, Salvia, Boussigaultia and Litchi can be employed. Saponins exert a positive effect on the function of the heart direct, or they help treat related diseases. For instance, they inhibit the formation of lipid peroxides in the cardiac muscle or in the liver, they influence the function of enzymes contained in them, they decrease blood coagulation, cholesterol and sugar levels in blood, they stimulate the immunity system. They act either direct, blocking the transfer of Ca2+ ions or modulating the function of Na(+)-K(+)-ATPase, or they help resorb other active principles. It can be concluded that saponins are a prospective group of drugs of natural origin for the prevention and treatment of diseases of the heart and circulatory system.

Cardiovascular Diseases↗

Inhibition of nifedipine-induced proliferation of cultured human gingival fibroblasts by Saiko, a Chinese herbal medicine.

Saiko is predominantly contained in Saireito, a Chinese herbal medicine. The present study was conducted to determine whether or not Saiko is involved in the inhibition by Saireito of nifedipine-induced proliferation and collagen synthesis in gingival fibroblasts. Nifedipine (10 microM) significantly enhanced the proliferation starting on day 5 of the culture period. When added together with nifedipine, Saiko at concentrations of 0.05%-0.2% (w/v) dose-dependently inhibited the nifedipine-induced proliferation, and at the highest concentration tested (0.2%), Saiko inhibited the nifedipine-induced proliferation by about 40%. Moreover, Saiko (0.2%) also inhibited the normal proliferation at days 11 and 14. Sole application of nifedipine (10 microM) augmented the release of bFGF, and Saiko concentration-dependently reduced the level of bFGF in the nifedipine-containing culture medium. Nifedipine (10 microM) increased the production of type I collagen to almost twice that of the control (normal medium), and Saiko at concentrations above 0.1% significantly reduced the nifedipineinduced production of collagen. In conclusion, the present findings demonstrate that Saiko inhibited the nifedipine-induced proliferation of gingival fibroblasts by reducing the release of bFGF and that Saiko is involved in the Saireito-induced inhibition of nifedipine-stimulated proliferation and collagen synthesis in gingival fibroblasts.

Bupleurum↗

Visual detection of saikosaponins by on-membrane immunoassay and estimation of traditional Chinese medicines containing Bupleuri radix.

The purpose of this study was to describe the simple, rapid, and environmental-cost effective determination method for saikosaponins in complicated samples like Bupleuri radix and traditional Chinese medicines (TCM). Saikosaponin standards, extracts of Bupleuri radix and TCM, were applied to a polyethersulphone (PES) membrane and developed by acetonitrile-water (1:4, by volume). Saikosaponin a (SSa), SSc, and SSd were visually detected by an immunostaining method (called Eastern blotting technique) using a monoclonal antibody (MAb) against SSa. At least 62.5 ng of SSa, SSc, and SSd were clearly detectable individually. These coloring spot areas of saikosaponins on PES membrane were calculated by using the NIH Imaging software and three saikosaponins can be analyzed quantitatively between 62.5 ng and 1.0 microg. Saikosaponins in Bupleuri radix and TCM were determined and these results of SSa and total saikosaponin concentrations were in good agreement with those from the ELISA analysis.

Bupleurum↗

A novel analytical ELISA-based methodology for pharmacologically active saikosaponins.

A competitive enzyme-linked immunosorbent assay (ELISA) for saikosaponins was established using monoclonal antibody (MAb) 3G10. Hybridoma 3G10 prepared by fusing splenocytes immunized with saikosaponin a-BSA (SSa-BSA) conjugate and a hypoxanthine-aminopterin-thymidine (HAT)-sensitive mouse myeloma cell line, P3-X63-Ag8-U1, secreted monoclonal antibodies with wide cross-reactivity to saikosaponins including saikosaponin b(2) (SSb(2)), c (SSc) and d (SSd), which are stereo and/or regio isomers of SSa. The method, at an effective measuring range of 0.6 mug /ml to 2.3 mug/ml of SSa, successfully detected total saikosaponins in Bupleuri radix and Kampo medicines prescribed with Bupleuri radix. Good correlation between ELISA and HPLC analyses of total saikosaponin in a crude extract of Bupleuri radix was obtained after hydrolysis of acyl saikosaponins by treatment with a mild alkaline solution.

Animals↗

Dual effect of saikogenin D: in vitro inhibition of prostaglandin E2 production and elevation of intracellular free Ca2+ concentration in C6 rat glioma cells.

To clarify the pharmacological profile of saikogenin D, we examined the effect of saikogenin D on prostaglandin E2 (PGE2) production and intracellular free Ca2+ concentration ([Ca2+]i) in C6 rat glioma cells. Saikogenin D (1-20 microM) inhibited PGE2 production induced by the Ca2+ ionophore A23187 in a concentration-dependent manner with the IC50 of about 3 microM. Saikogenin D did not affect the conversion of arachidonic acid into PGE2 in microsomal preparations. On the other hand, saikogenin D elevated [Ca2+]i in a concentration-dependent manner (10-100 microM) with the EC50 value of about 35 microM in the presence or absence of extracellular Ca2+. These results suggest that saikogenin D possesses a dual effect: an inhibition of A23187-induced PGE2 production without a direct inhibition of cyclooxygenase activity; and an elevation of [Ca2+]i that is attributed to Ca2+ release from intracellular stores.

Animals↗

Effects of chai hu (radix burpleuri) containing formulation on plasma beta-endorphin, epinephrine and dopamine on patients.

Chai Hu (Radix Burpleuri), a major ingredient in many traditional Chinese medicine formulas, such as Xiao Yan Wan, is used in the treatment of liver stagnation and spleen deficiency syndrome (LSSDS). The objectives of this study were to examine the effects of Xiao Yao Wan containing Chai Hu on the changes of plasma indices in patients with LSSDS. Fifty-eight cases of LSSDS were randomly divided into two groups: 41 cases in the experimental group were treated with Xiao Yao Wan containing Chai Hu and 17 cases in the control group were treated with Zhi Bai Di Huang Wan for one consecutive month in a single blind design. Before and after treatment, high performance liquid chromatography (HPLC) was applied to determine the changes of plasma norepinephrine (NE), epinephrine (E) and dopamine (DA). Radioimmunoassay was performed to measure the amount of plasma beta-endorphin (beta-EP), adrenocorticotropin hormone (ACTH), estradiol (E2) and testosterone (T), and laser nephelometry was also conducted to measure plasma immunoglobulin A (Ig A) and G (Ig G). Compared to baseline levels, plasma beta-EP was significantly increased (p < 0.01), while E and DA were markedly decreased (p < 0.01) after the administration of Xiao Yao Wan in the experimental group. The other indices did not change. This is the first evidence showing that the effect of Xiao Yao Wan containing Chai Hu on the treatment of patients with LSSDS may be through enhancing plasma beta-EP and decreasing E and DA release. We conclude that Xiao Yao Wan containing Chai Hu regulates nervous and endocrine systems and contributes to the improvement of the clinical status of patients with LSSDS.

Adult↗

Development of an assay system for saikosaponin a using anti-saikosaponin a monoclonal antibodies.

For immunization, saikosaponin a (SSa) was conjugated with bovine serum albumin (BSA). The hapten number in an antigen conjugate was determined to be eleven by matrix-assisted laser adsorption/ionization time-of-flight mass spectrometry (MALDI-TOF Mass). Hybridomas secreting monoclonal antibodies (MAb) against SSa were produced by fusing splenocytes immunized with SSa-BSA conjugate and a hypoxanthine-aminopterin-thymidine-sensitive (HAT) mouse myeloma cell line, P3-X63-Ag8-653. A high specific MAb against SSa was selected from hybridomas using enzyme-linked immunosorbent assay (ELISA) analysis. Weak cross-reactivities occurred with saikosaponin c, b(2) and d, which are stereochemical and/or functional isomers of SSa, but no cross-reactivities were observed with other related steroidal glycosides. The full range of the assay extends 26 ng/ml to 1.5 microg/ml of SSa. Good correlation of SSa concentrations in a crude extract of Bupleuri radix between ELISA and HPLC methods was obtained after hydrolysis of acyl saikosaponins by treatment with a mild alkaline solution. The newly established ELISA has been applied for the quantitative assay of SSa in the Bupleuri radix and the Kampo medicines (TCM) prescribed with Bupleuri radix.

Animals↗

High performance liquid chromatographic assay of saikosaponins from radix Bupleuri in China.

In this study, the quantitative analysis of saikosaponins from Radix Bupleuri in China was performed by high performance liquid chromatography. Saikosaponin-a and -d were converted completely into saikosaponin-b1 and -b2 by mild acid treatment. Distinctive measuring of these converted diene-saponins provided a rapid and selective method for the determination of saikosaponin-a and -d in commercial samples of Radix Bupleuri. The conditions of extraction and conversion of saikosaponins were optimized using orthogonal design L9(3(4)). The HPLC analysis was performed on ODS-C18 column with a flow rate of 1.0 ml/min and detection wavelength of 250 nm. Well resolved chromatograms of saikosaponin-b1 and -b2 were obtained with an isocratic elution of acetonitrile : 1% formic acid water (37.5 : 62.5). Calibration curves of saikosaponin-b1 and -b2 were linear in the range of 4.9-98.0 microg/ml and 3.5-71.0 microg/ml, respectively. The average recovery of saikosaponin-b1 and -b2 were 98.3% (RSD = 3.1%) and 96.4% (RSD = 1.8%), respectively. Seventeen samples of different species and habitats of Radix Bupleuri were analyzed by the developed HPLC method.

Bupleurum↗

Analysis of the essential oil from radix bupleuri using capillary gas chromatography.

A simple and rapid capillary gas chromatographic (CGC) method with flame ionization detection has been newly developed for analysis of the essential oil from Radix Bupleuri. Twenty components were identified with gas chromatography-mass spectrometry. E-2-heptenal, furan, 2-pentyl, and E-2-nonenal were quantified simultaneously using the internal standard method. Decane was used as an internal standard. Separation and quantification were achieved on a DB-5 capillary column (30 m x 0.25 mm i. d., 0.25-microm film thickness). The oven temperature was programmed as follows: 60 degrees C to 70 degrees C at 1 degree C/min rate, 70 degrees C for 10 min, 3 degrees C/min to 120 degrees C, 20 degrees C/min to 250 degrees C, and held at 250 degrees C for 5 min. The oven pressure was programmed as follows: 46.1 kPa for 25 min, 20.0 kPa/min to 77.6 kPa, and then held for 22 min. Split injection was conducted with a split ratio of 10:1; flow-rate, 1.00 ml/min; carrier gas, nitrogen; injector temperature, 280 degrees C; and detector temperature, 280 degrees C. The system proved effective in resolving E-2-heptenal, furan, 2-pentyl, and E-2-nonenal peaks from their interfering components. The method displayed excellent linearity in the range of 26.8-1072 microg/ml (E-2-heptenal), 6.5-1292 microg/ml (furan, 2-pentyl), and 7.8-1564 microg/ml (E-2-nonenal). The average recovery rates of E-2-heptenal, furan, 2-pentyl, and E-2-nonenal were 100.3%, 102.8%, and 97%, respectively. CGC is a quick and accurate method for analysis of the essential oil from Radix Bupleuri.

Bupleurum↗

Adult respiratory distress syndrome induced by a Chinese medicine, Kamisyoyo-san.

We report the first case of Kamisyoyo-san-induced adult respiratory distress syndrome (ARDS). A 59-year-old female was given Kamisyoyo-san for treatment of seborrheic dermatitis. She then presented with a respiratory illness having clinical, radiologic and functional characteristics of ARDS. Bronchoalveolar lavage fluid showed an increased number of lymphocytes, neutrophils and eosinophils. The lymphocyte stimulation test with Kamisyoyo-san was positive.

Bupleurum↗

[Chemical constituents analysis on anticonvulsive effect of three extracts from radix bupleuri].

OBJECTIVE: To study the relationship between anticonvulsive effect of three extracts from Radix Bupleuri and their chemical compositions. METHODS: Volatile oil component, saikosaponin component and water-soluble component were extracted with SFE-CO2 from Radix Bupleuri, their effect on MES and PTZ seizure models had been studied, and their compositions were analyzed by GC-MS and HPLC. RESULTS: Compared with the control, volatile oil component and saikosaponin component significantly decreased anticonvulsive occurrence on MES, the main chemical compositions of them was linoleic acid; water-soluble component significantly decreased anticonvulsive occurrence on PTZ seizure, but most of its compositions were glycerol and xylitol. CONCLUSION: Linoleic acid from Radix Bupleuri maybe fight against MES seizure, while glycerol and xylitol could fight against PTZ seizure.

Anticonvulsants↗

[The effect of he jie decoction on the TCRV beta 7 of chronic hepatitis B patients].

OBJECTIVE: To explore the correlation of clinical effect and TCRV beta 7 expression of He Jie Decoction (HJD) on chronic hepatitis B patients. METHODS: 45 patients of chronic hepatitis B were divided randomly into two groups. 30 patients in the treated group were treated by HJD, and 15 patients in the control group were treated by usual western medicine. The TCRV beta 7 level were detected before and after being treated. RESULTS: After six months treatment, the ALT level of two groups were obviously decreased (P < 0.01), TCRV beta 7 expression were detected out in 6 patients of the treated group, and HBV-DNA and HBeAg of the 6 patients were negative conversion. The TCRV beta 7 expression couldn't be detected out in the control group, and HBV-DNA and HBeAg of the control group weren't negative conversion. There was no significant difference between the clinical total effective rate of the two groups (P > 0.05), but the apparent rate of the treated group was significantly higher than that of the control group (P < 0.01). CONCLUSION: HJD might have the effect on improving the TCRV beta 7 expression of chronic hepatitis B patients, which might be the ways of HJD inhibiting and killing hepatitis B virus.

Adolescent↗