The Identification of Phenylalanine Ammonia-Lyase(PAL)Genes from Pinus yunnanensis and an Analysis of Enzyme Activity in vitro

Phenylalanine ammonia lyase(PAL)is the rate-limiting and pivotal enzyme of the general phenylpropanoid path-way,but few reports have been found on PAL genes in Pinus yunnanensis.In the present study,three PAL genes were cloned and identified from P.yunnanensis seedlings for thefirst time,namely,PyPAL-1,PyPAL-2,and PyPAL-3.Our results indicated that the open-reading frames of PyPAL genes were 2184,2157,and 2385 bp.Phylogenetic tree analysis revealed that PyPALs have high homology with other known PAL genes in other plants.In vitro enzymatic analysis showed that all three PyPAL recombinant proteins could catalyze the deamination of L-phenylalanine to form trans-cinnamic acid,but only PAL1 and PAL2 can catalyze the conversion of L-tyrosine toρ-coumaric acid.Three PyPAL genes were expressed in different tissues in 1-year-old P.yunnanensis,and such genes had different expression patterns.This study lays a foundation for further understanding of the biosynthesis of secondary metabolites in P.yunnanensis.

Changes in Polyphenols and Antioxidant Activities of Yingshan Yunwu Tea during Digestion in Vitro

[Objectives]To explore the change rule of polyphenol content and antioxidant activity of coarse old leaves of Yingshan Yunwu Tea in the process of human digestion.[Methods]The coarse and old leaves of Yunwu tea in Yingshan,Huanggang,Hubei Province were selected as the research object,and their digestion in vitro was simulated.The total polyphenol content was determined by Folin-phenol reagent colorimetric method,and the DPPH radical scavenging activity and total antioxidant activity were determined.[Results]After simulated gastrointestinal digestion in vitro,the polyphenol content and antioxidant activity of coarse old leaf tea soup showed a downward trend.After gastrointestinal digestion,the polyphenol content in tea infusion separately decreased by 31.8%and 8.5%;the scavenging rate of DPPH free radical was 97%before digestion,decreased to 92%after gastric digestion and 65%after intestinal digestion,which decreased by 5%and 27%,respectively;after gastrointestinal digestion,the total antioxidant capacity of tea soup decreased by 4.7%and 3.1%,respectively.[Conclusions]This study provided a reference for the development and application of coarse old leaves of Yingshan Yunwu tea,and provided a reference for the nutritional value evaluation and comprehensive utilization of coarse old leaves,so as to make the best use of coarse tea leaves and reduce the waste of resources.

A Study on Anti-oxidative Activity of Soybean Peptideswith Linoleic Acid Peroxidation Systems

Soybean bioactive peptides（SBPs） were prepared from the isolated soybean protein by proteolysis with an alkaline protease, alcalase, at 50℃ and pH = 8.0. The dependence of hydrolysis time on hydrolysis degree and molecular weight distribution were examined. The hydrolysate was fractionated on a Sephadex G-25 column and the anti-oxidative activities of the fractions were detected by the method of pyrogallol auto-oxidation. The average chain length of soybean peptides that have anti-oxidative activity was estimated to be about 7. The anti-oxidative properties of the soy- bean peptide were also studied by using linoleic acid peroxidation systems. The optimal condition of the peroxidation system was set up, Vc/Cu^2 ＋ as the inducer at pH = 7.4 and 25 ℃. In addition, soybean peptides show higher antioxidative activity compared with GSH.

In vitro antioxidant and antimicrobial activity cycloart-23-ene-3β,25-diol(B2) isolated from Pongamia pinnata(L.Pierre)

Objective:To evaluate the in-vitro antioxidant and antimicrobial activity of cvcloart-23- ene-3β,25-diol(called as B2) isolated from stem bark of Pongamia pinnata.Methods:In vitro antioxidant activity of B2 was determined by methods for determination of Dl’PH radical scavenging,reducing power,superoxide anion radical scavenging,hydroxyl radical scavenging, hydrogen peroxide scavenging,metal chelating and nitric oxide radical scavenging at the doses of 20.40.60.80 and 100μg/mL,respectively.3 -tocopherol with same concentration was used as a standard antioxidant.In vitro antimicrobial activity of B2 was determined by cup plate method in different concentration range of 10-100μg/mL.Results:The results indicated that dose dependent%reduction against DPPH radical,reducing power,superoxide anion radical scavenging,hydroxyl radical scavenging,metal chelating,hydrogen peroxide scavenging and nitric oxide radical scavenging by B2 andβ-tocopherol.Conclusions:It is concluded that cycloart 23-ene-3β,25 diol(B2) showed dose dependent antioxidant activity.B2 showed more DPPH radical scavenging,reducing power,superoxide scavenging,hydroxyl radical scavenging, metal chelating scavenging,hydrogen peroxide radical scavenging and nitric oxide radical scavenging activity thanβ-tocopherol and in case of antimicrobial activity B2 exhibited broad-spectrum activ ity against bacteria and strong activity against yeast type of fungi.

In vitro Inhibitory Activity of Shisandra chinensis and Polygonatum sibiricum against Vibrio harveyi and Its Biofilms

[Objective] This study aimed to analyze the in vitro inhibitory activity of Shisandra chinensis and Polygonatum sibiricum against Vibrio harveyi and its biofilms. [Result] By agar diffusion test, in vitro inhibitory activity of 5. chinensis and P. sibiricum against V. harveyi was investigated. The minimal inhibitory concentration （ MIC） and minimal bactericidal concentration （MBC） of 5. chinensis and P. sibiricum against V. harveyi were determined by doubling dilution meth-od. The inhibitory activity of 5. chinensis and P. sibiricum on the formation of V. harveyi biofilms was evaluated by modified MTT assay. [ Result ] Both 5. chinen-sis and P. sibiricum had inhibitory activity against V. harveyi. The inhibition zone diameter of 5. chinensis against V. harveyi was 17. 95 mm; MIC and MBC of 5. chinensis were both 3.125 mg/ml. The inhibition zone diameter of P. sibiricum against V. harveyi was 12. 22 mm; MIC and MBC of P. sibiricum were 3.125 and 6.250 mg/ml, respectively. When the concentration was higher than 6. 25 mg/ml, 5. chinensis decoction had extremely significant inhibitory activity against V. harveyi （P 〈 0. 01） ; when the concentration was higher than 3. 125 mg/ml, P. sibiricum had extremely significant inhibitory activity against V. harveyi （P 〈0. 01）. [ Conclusion] 5. chinensis and P. sibiricum could significantly inhibit V. harveyi and its biofilms.

Effect of Gamma Irradiation on Total Phenolic Content and <i>in Vitro</i>Antioxidant Activity of Pomegranate (<i>Punica granatum</i>L.) Peels

Pomegranate peels were studied for the effect of gamma irradiation on microbial decontamination along with its effect on total phenolic content and in vitro antioxidant activity. Gamma irradiation was applied at various dose levels (5.0, 10.0, 15.0 and 25.0 kGy) on pomegranate peel powder. Both the values of total phenolic content and in vitro antioxi- dant activity were positively correlated and showed a significant increase (p < 0.05) for 10.0 kGy irradiated dose level immediately after irradiation and 60 days of post irradiation storage. At 5.0 kGy and above dose level, gamma irradia- tion has reduced microbial count of pomegranate peel powder to nil. Post irradiation storage studies also showed that, the irradiated peel powder was microbiologically safe even after 90 days of storage period.

Antiviral Activity of GuiQi Polysaccharides against Enterovirus 71 in vitro

In this study,we have investigated the antiviral activity of GuiQi polysaccharides (GQP) upon enterovirus 71 (EV71) in vitro.An assay using methyl thiazolyl tetrazolium (MTT),and analyses of cytopathic effects (CPE)were used to examine the antiviral activity of GQP upon Vero cells infected with EV71.The results revealed that GQP at concentrations below 31.2μg/mL exhibited significant antiviral effects upon EV71 when applied under three different experimental protocols.GQP was most strongly active in preventing the adsorption of EV71 to target cells and in this respect it was significantly more effective than ribavirin.In addition,it was clear that GQP could inhibit viral replication when added to cells 2 h after infection,but if added at the point of infection its effect was weak.GQP is considered to be less toxic than ribavirin,and may warrant further evaluation as a possible agent in the treatment of hand,foot and mouth disease (HFMD).

Potential bone-inducing activity in vitro of recombinant human bone morphogenetic protein-7 from a CHO expression system

Objective: To express the recombinant human bone morphogenetic protein-7(rhBMP-7) in Chinese hamster ovary(CHO) cells, and to establish the in vitro biological activity assay of rhBMP-7.Methods: Human BMP-7 cDNA was subcloned into p114 mammalian expression vector and transfected to CHO cells by using the Lipofectamine 2000 transfection method. CHO cell supernatants were harvested and analyzed to identify the molecule mass of secreted rhBMP-7 and examine its biological activity in vitro to stimulate the synthesis of alkaline phophatase(ALP), a characteristic of osteoblast phenotypes. Results: rhBMP-7 was produced stably in CHO cells, as a processed mature disulfide-linked homodimer, with an apparent molecular mass of 36 000. Examination of the rhBMP-7 biological activity showed that rhBMP-7 specifically stimulated the production of ALP(4-fold increase at 100 ng of rhBMP-7/ml). Conclusion: The rhBMP-7 from CHO expression system has significant biological activity in induction of osteoblast phenotype, which demonstrates rhBMP-7 has the potential bone regeneration activity.

Rabbit Defensin(NP-1) Gene Transformation of Wheat and In Vitro Microbicidal Activity Assay of Transgenic Plants

Monocot high expression vector pBARUNP1, harboring rabbit defensin(NP 1) gene and selective bar gene for resistance to the herbicide Basta, were constructed and then transferred into immature embryos of wheat (“Bobwhite” and “Zhong 60634”)via particle bombardment. Southern and RNA dot blots showed the stable integration and transcription of foreign NP 1 gene in the wheat genome. Furthermore, in vitro microbicidal activity assay indicated the proper translation of defensin. Crude protein extraction of transgenic plants exhibited to some extent cytotoxic to several pathogens including G. saubinetii, B. subtilis, E.coli, and A. tumefaciens.

Research progress of in vitro activation mechanism and clinical application of female ovarian tissue

The activation and development of primordial follicles is the key to the maturation of female gametes.Premature ovarian insufficiency(POI)patients are unable to complete the primordial follicle activation and development due to follicular dormancy and unbalanced developmental regulation in the body,leading to female infertility.Ovarian tissue in vitro activation(IVA)technology has become a new way to solve the problem of patients who cannot auto-activate primordial follicles to obtain their own mature oocytes.In IVA research,signaling pathways such as PI3K/PTEN/Akt and Hippo have become the focus of current research.This review will describe the relevant research progress and clinical application of the IVA mechanism,and provide a reference for clinical research on ovarian tissue culture and activation in vitro.

In Vitro Anti-Bacterial and Anti-Fungal Activities of Extracts from Different Parts of 7 Zingiberaceae Plants

This study aimed to explore the anti-bacterial and anti-fungal activities of extracts from different parts of plants in the Zingiberaceae family.The inhibitory rate,minimum inhibitory concentration(MIC),and minimum bactericidal concentration(MBC)of leaf and stem,and root and rhizome extracts from Alpinia katsumadai Hayata,Alpinia oxyphylla Miq×Alpinia henryi K.Schumann,Alpinia oblongifolia Hayata,Alpinia nigra(Gaertn.)Burtt,Amomum villosum Lour,Alpinia zerumbet(Pers.)Burtt.et Smith and Alpinia oxyphylla Miq were determined using the fungus cake method and double dilution method.The seven Zingiberaceae plants exhibited characteristic antibacterial activities against pathogenic bacteria and fungi.At a 1.5 mg mL^(−1),A.zerumbet root and rhizome extracts exhibited strong inhibitory activity against S.aureus and E.coli,with 83.23%and 79.62%,respectively.In addition,A.zerumbet leaf and stem extracts had an inhibitory rate of 90.85%against P.aeruginosa.At the same concentration,the leaf and stem,root and rhizome extracts of A.katsumadai had the best anti-bacterial effect against F.oxysporum,with inhibition rates of 84.46%and 84.73%,respectively.Moreover,A.katsumadai and A.zerumbet leaf and stem extracts had the most significant antibacterial effect against S.aureus,with a MIC of 0.063 mg mL^(−1).Thus,both A.katsumadai and A.zerumbet extracts had significant antibacterial activity.In addition,by comparing the inhibitory effect of extracts from different parts,it was found that the inhibitory rate and average inhibitory rate of extracts from leaf and stem were higher than those from root and rhizome.The chemical constituents of A.katsumadai and A.zerumbet,determined by the high-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS),revealed that citric acid(CA),alpinetin,and pinocembrin(PNCB)were the functional constituents yielding the antibacterial activity.Overall,A.katsumadai and A.zerumbet have the potential to be developed as new plant fungicides and bactericides.

In vitro antibacterial activity of Hibiscus rosa-sinensis flower extract against human pathogens

Objective:To access the in vitro antibacterial activity of Hibiscus rosa-sinensis(H.rosa-sinensis) flower extract against human pathogens.Methods:Antibacterial activity was evaluated by using disc and agar diffusion methods.The protein was run through poly acrylmide gel electrophoresis to view their protein profile.Results:The results showed that the cold extraction illustrates a maximum zone of inhibition against Bacillus suhtillis(B.suhtillis),Escherichia coli(E.coli) viz.,(17.00±2.91),(14.50±1.71)mm.followed by hot extraction against.E.coli.Salmonella sp.as(11.66 ±3.14),(10.60±3.09)mm.In methanol extraction showed a highest zone of inhibition recorded against B.suhtillis,E.coli as(18.86±0.18),(18.00±1.63) mm pursued by ethanol extraction shower) utmost zone of inhibition recorded against Salmonella sp.at(20.40±1.54) mm.The crude protein from flower showed a maximum inhibitory zone observed against Salmonella sp.,E.coli viz.,(16.55±1.16),(14.30±2.86)mm.The flower material can be taken as an alternative source of antibacterial agent against the human pathogens.Conclusions:The extracts of the H.rosasinensis art;proved to have potential antibacterial activity,further studies arc highly need for the drug development.

Evaluation of in-vitro antibacterial activity and anti-inflammatory activity for different extracts of Rauvolfia tetraphylla L.root bark

Objective:To assess the in-vitro antihacterial activity and anti-inflammatory activity of orally administered different extracts(Hydro-alcoholic,methanolic,ethyl acetate and hexane)of Rauvolfia tetraphylla(R.tetraphylla)root bark in Carrageetiaii induced acute inflammation in rats.Methods:In-vitro antibacterial activity was evaluated for extracts against four Gram positive and four Gram negative bacteria by using cylinder plate assay.Hydro-alcoholic extract(70%v/v ethanol)at 200,400 and 800 mg/kg doses and methanolic,ethyl acetate and hexane extracts at doses 100,200 and 400 mg/kg were tested for anti-inflammatory activity in Carrageenan induced rat paw oedema model and paw thickness was measured every one hour up to 6 hrs.Results:All extracts of R.tetraphylla root bark showed good zone of inhibition against tested bacterial strains.In Carrageenan induced inflammation model,hydro-alcoholic and methanolic extract of R.tetraphylla root bark at three different doses produced significant(P<0.00l)reduction when compared to vehicle treated control group and hexane,ethyl acetate extracts.Conclusions:In the present study extracts of R.tetraphylla root bark shows good in-vitro antibacterial activity and in-vivo anti-inflammatory activity in rats.

Representativeness of EN 1040/13727 Assay Conditions for Evaluating <i>In Vitro</i>the Bactericidal Activity of a Chlorhexidine Digluconate and Benzalkonium Chloride Antiseptic Preparation

Aims: The representativeness of the mandatory bacterial strains specified in European standards for in vitro assay of the bactericidal activity of antiseptics was evaluated by testing the activity of an antiseptic combining chlorhexidine digluconate 0.2% and benzalkonium chloride 0.5% against 21 additional bacterial strains, and the positive interaction between these two biocidal agents was assessed. Methods and Results: The bactericidal activity of the antiseptic solution used pure or diluted was assessed according to the European standards EN 1040 and EN 13727. The contact time was 1 min at 20°C. Interfering substances used in the EN 13727 assay were bovine serum albumin and sheep erythrocytes, simulating “dirty” conditions, and hard water. A reduction of colony-forming units by ≥5 log10 was deemed to meet the requirements to conclude bactericidal activity. Under “basic” conditions, the bactericidal activity of the antiseptic was observed against all four mandatory strains specified in the standards as well as against nearly all the additional strains tested, including most of those with acquired antibiotic-resistance. The positive interaction between the two biocidal agents was also confirmed. Under “dirty” conditions, the bactericidal activity of the antiseptic solution was maintained against all the mandatory strains and was reduced against only four of the additional strains tested. Conclusions: With regard to the antiseptic tested and under the experimental conditions described, bactericidal activity evidenced against the mandatory strains appeared to be representative of that manifested against a wide range of the main pathogenic bacteria. Reduced bacterial activity against some of the additional strains tested (e.g. Enterobacteriaceae) was observed under “dirty” conditions. Significance and Impact of the Study: EN 13727 with some experimental adjustments represents an additional appropriate standard that needs to be considered for mucocutaneous antiseptic assessment. However, it may be worth including other specific bacterial strains to those specified in the standard, when evaluating antiseptics intended for use in certain clinical situations.

Antifilarial activity of ethyl acetate extract of Vitex negundo leaves in vitro

Objective:To evaluate the possible antifilarial effect of ethyl acetate extract of Vitex negundo(Verhenaceae)leaves against Selaria cervi filarial parasite in vitro.Methods:In vitro screening was done by the method of motility inhibition and NTT reduction assay with concentralions of0.03 to 1.00 mg/mL for 2 to 24 h incubation periods respectively,for possible antifilarial effect by comparing with control.Results:In motility assay,complete inhibition of motility was observed and in MTT reduction assay which gave>50%reduction for concentrations 0.20,0.50and 1.00 mg/mL at 10,6 and 2 h incubation periods respectively in a dose dependent manner(P<0.05).An antifilarial effect imparted by plant extract was found to be a function of their relative concentrations.Inhibitory concentration(IC_(50))for the plant extract was found to be 0.16mg/mL.Conclusions:The present study recorded significant antifilarial effect of Vitex negundo plant extract and contributed toward the development of database for novel drug candidates for lymphatic fllariasis.

In vitro Antioxidant Activity of Leontopodium franchetii Beauv.

[Objectives] To study the in vitro antioxidant activity of L. franchetii Beauv. [Methods] The DPPH and ABTS^+ scavenging rate and reducing power of L. franchetii Beauv. were detected by ultraviolet spectrophotometry. [Results] The 70% ethanol extract of L. franchetii Beauv. had strong antioxidant capacity, and its antioxidant capacity increased with the increase of mass concentration. [Conclusions] The 70% ethanol extract of L. franchetii Beauv. has higher DPPH and ABTS^+ scavenging rate and reducing power.

Spatiotemporal Neural Activity Changes in the Molluscan Olfactory Center Specifically Induced by Innately Aversive and <i>In Vitro</i>Aversively Conditioned Odors

In the procerebrum (PC), the olfactory center, of the land slug Limax, an oscillation of local field potential (LFP) with 0.5 - 1 Hz is observed by electrophysiological extracellular recording. The oscillation has a phase delay along the distal-proximal axis, resulting in the propagation of waves from the distal to proximal region. One important advantage of nervous systems of mollusks such as Limax is that their nervous systems in vitro retain several types of computational properties found in vivo (e.g. learning and memory). A previous study showed that the LFP frequency in the PC of Limax increased specifically in response to innately aversive and in vitro aversively conditioned odors. In the present study, we examined spatiotemporal neural activity changes induced in the PC by those odors using the fluorescent voltage imaging technique. The results showed that innately aversive (onion and hexanol) and in vitro aversively conditioned (carrot, which is innately attractive) odors specifically induced an increase in propagation speed of the neural activity in the PC, while innately attractive odors did not induce it. The results also suggested that the avoidance behavior by those odors might be induced by the increase of propagation speed and the following increases in the discharges of the partial nerve that transmits the motor output.

<i>In Vitro</i>Evaluation of Ozone Activity on Recent Clinically Isolated Bacterial Strains

This study aims to evaluate the cozone bactericidal activity in different suspension media (saline, broth and whole blood) at different exposure times. Methicillin-resistant Staphylococcus aureus, Enterococcus faecalis, ESBLpositive Escherichia coli, MDR Pseudomonas aeruginosa were suspended in different media. We used a bacterial concentration of 0.2 MF for all experiments, as this concentration is consistent with the results of septic shock blood experiments. We performed ozone insufflations in a “sealed environment”. The total number of insufflations for each experiment ranged from one to four. The gas concentration was maintained at 80 mcg/ml. We confirmed the bactericidal activity of ozone on saline for all the bacterial strains. Experiments in broth revealed no changes in the bacterial growth. Ozone is primarily bactericidal against E. coli and bacteriostatic on P. aeruginosa, S. aureus and E. faecalis on whole blood. This study confirms the bactericidal efficacy of topical ozone applications and supports the need for further evaluations of the therapeutic potential of major ozone autohemotherapy. The results in E. coli promote further investigations of ozone activity on other Enterobacteriaceae and its potential use in the treatment of urinary infections. In general, these results suggest that ozone-therapy might be an alternative therapy to overcome antibiotic resistance.

IN VITRO ANTITUMOR ACTIVITY OF TUMOR-INFILTRATING LYMPHOCYTES FROM HUMAN GASTRIC CARCINOMA

Tumor-infiltrating lymphocytes (TIL) isolated from 11 gastric carcinoma were studied. TIL could grow for a long-term in medium containing recombi-nant interleukin-2(rlL-2). The mean expansion fold achieved in 6 long-term cultures of 11 specimens was 15.1. RIL-2 expanded gastric TIL exhibited significant cytotoxicity against K562, BGC823, MCF-7 and more effective antitumor cytotoxicity against fresh autologous tumor targets and human gastric cancer cell line. Peak cytotoxicity was shown in the third or fourth week after cultures. Cryopreservation of gastric TIL didn't influence their expansion capacity and antitumor activity. Phenotypic analysis was demonstrated in this study. The results of present study indicate that TIL from human gastric carcinoma could be expanded and reach high levels of antitumor effector function in long-term cultures with rIL-2. Their function may be of clinical importance.

Effects of Buyang Huanwu decoction and Astragalus mongholicus on platelet activating factor receptor activity in rabbits in vitro

BACKGROUND: The pharmacological action of traditional Chinese medicine compound is the comprehensive effect of the various ingredients, and the interactions of various ingredients are closely correlated with the final effect. In order to reveal the compatibility mechanism of BHD's prescription in treating and preventing ischemic cerebrovascular disease, we needed explore the effect and relation of ingredients in the prescription. OBJECTIVE: To observe the effect of Buyang Huanwu decoction (BHD) and Astragalus mongholicus on the activity of platelet activating factor receptor (PAFR) in the platelet of rabbits in vitro, and investigate the mechanism of Astragalus mongholicus. DESIGN: A decomposed recipes study. SETTING: Guangzhou University of Traditional Chinese Medicine. MATERIALS: Five New Zealand rabbits, weighing 2-3 kg, both sexes, were used. BHD was composed of Sheng Huang Qi 120 g, Dang Gui Wei 6 g, Chi Shao 4.5 g, Chuan Xiong 3 g, Di Long 3 g, Tao Ren 3 g, Hong Hua 3 g. The prescription for activating blood circulation consisted of Dang Gui Wei 6 g, Chi Shao 4.5 g, Chuan Xiong 3 g, Di Long 3 g, Tao Ren 3 g and Hong Hua 3 g. The prescription for invigorating qi consisted of 120 g Sheng Huang Qi. The prepared herbal pieces were purchased from the traditional Chinese medicine Dispensary of Foshan Second People's Hospital, and appraised by Professor Xu from Science of Chinese Materia Medica College, Guangzhou University of Traditional Chinese Medicine. 3H-PAF was supplied by Amersham Co., Ltd. (specific activity: 6. 475 TBq/mmol; batch number: 200402); PAF standard by Biomol Co., Ltd. (batch number: P1318V). METHODS: The experiments were carried out in the Laboratory of Nuclear Medicine, Guangzhou University of Traditional Chinese Medicine from September to December 2004. ① Injections of BHD, prescriptions for activating blood circulation and invigorating qi were prepared by the decoction and alcohol sedimentation technique. Rabbit common carotid artery blood (40 mL) was drawn via intubation to prepare platelet suspension of (0.8-1.0)×1010 L-1. ② Determination of 3H-PAF and washed PAFR binding: The general combination tube (T) contained washed platelet-rich plasma (WPRP) 380 μL + 3H-PAF (0.35 nmol/L)10 μL+distilled water 5 μL; The nonspecific binding tube (P) contained WPRP 380 μL+3H-PAF(0.35 nmol/L)10 μL+cold PAF (1 μmol/L) 5 μL; The sample tube (Y) contained WPRP 380 μL+3H-PAF(0.35 nmol/L)10 μL+experimental medicine (injection of BHD, prescriptions for activating blood circulation or invigorating qi) 5 μL. The test was conducted for three times for each sample in the same way as mentioned above. The samples were shaken on the oscillator for 30 s, then bathed at 25 ℃ for 40 minutes, and the reaction was terminated with cold Tris buffer containing 0.1% BSA, multichannel cell detachment separator was used for vacuum suction to filter the separated free 3H-PAF, and the filter paper was washed with cold Tris buffer for four times, then dried in the baking oven (80 ℃) for 1 hour, and placed in xylol liquid scintillator, and the radioactivity was determined automatically by the liquid scintillation detector. The mean of the three parallel tubes was calculated. The specific binging inhibition rate was calculated: SBIR=[(T-Y)/(T-P)]×100%]. ③ Univariate analysis of variance was conducted. And for comparison of each paired groups, the q test was adopted. MAIN OUTCOME MEASURES: Effect of BHD whole prescription, prescriptions for activating blood circulation and invigorating qi on the specific binding inhibition rate of 3H-PAF and PAFR. RESULTS: BHD, prescriptions for activating blood circulation and invigorating qi were all able to inhibit the specific binding of 3H-PAF to PAFR, the specific blinding inhibition rates were (45.90±7.50)%, (97.90±1.84)% and (26.75±2.48)%, respectively, and there were significant differences between every two groups (P < 0.01). CONCLUSION: Single Astragalus mongholicus (120 g) can inhibit the specific blinding of PAFR in the platelet of the rabbit with 3H-PAF, but the combination of Astragalus mongholicus with the drugs for activating blood circulation in BHD can significantly decrease the inhibiting action of the latter on PAFR activity of the platelet, reflecting the combined mechanism of 'removing blood stasis without injuring the vital qi' in BHD.