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OBJECTIVE To optimite the purification technology of total triterpenoid extracts from Inonotus obliquus ，and to investigate the anti -tumor activity of its purified products . METHODS Using inotodiol as control ，the method was established for the content determination of total triterpenoid in I. obliquus. The type of macroporous adsorption resin ，sample volume ，sample concentration，sample flow rate ，eluent volume ，eluent dosage and elution flow rate were selected by single factor experiments . The purification technology of the crude extract was determined and verified . The effects of total triterpenoid purified from I. obliquus on the proliferation ，migration and apoptosis of human cervical cancer HeLa cells were detected by cell proliferation test ， migration test ，flow cytometry and AO/EB kit . RESULTS The best purification technology of total triterpenoid crude extracts from I. obliquus was as follows ：AB-8 macroporous adsorption resin was used ；mass concentration of the sample solution was 2.0 mg/mL；sample volume was 140 mL，and the flow rate was 1.0 mL/min；the impurity was removed with 50% ethanol 40 mL， then eluted with 95% ethanol 160 mL，at the elution flow rate of 3.0 mL/min. After purification ，mass concentration of total triterpenoid from I. obliquus increased from 34.36% to 73.39%. The total triterpenoid of I. obliquus could inhibit the proliferation of HeLa cells ，and the 50% inhibitory concentration was 184.20 μg/mL. Compared with control group ，the purified products could significantly inhibit the migratio n and promote the apoptosis of HeLa cells （P＜0.05 or P＜0.01）. CONCLUSIONS The purification technology of total triterpenoids extracts from I. obliquus is successfully optimited . The purified product could inhibit the proliferation and migration of HeLa cells and induce their apoptosis.
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Objective To explore the effect of Cortex Eucommiae(CE)extract on oxidative stress markers of brain aging mice. Methods Brain aging mice model was made via subcutaneously administered D-galactose. The ethanol extract of CE was intra?gastrically administered to the model mice. Apoptosis ratio of brain cells of mice was determined by a flow cytometry to evaluate the an?ti-aging function of CE extract. A series of biomarkers related to oxidation,including malonyldialdehyde(MDA),superoxide dis?mutase(SOD),protein carbonyl(PCO),and nitric oxide(NO)in serum of mice were determined by ELISA. Results Compared with the model mice,the apoptosis ratio of brain cells in the CE extract group decreased significantly(P<0.01). ELISA test results showed that,compared with the normal group,NO and SOD levels in serum of CE extract group were significantly higher(P<0.01). Compared with the model group,MDA and PCO levels in serum of CE extract group were significantly lower(P<0.01),and SOD level was obviously higher(P<0.01). Conclusion The CE extract might play the role of brain anti-aging by the effective attenuation of ox?idative damage.
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OBJECTIVE:To study the effects of scorpion’s proteins with different molecular weights on angiogenesis of the transgenic zebrafish. METHODS:The vascular fluorescence transgenic zebrafish models were established. Scorpion’s proteins were separated by ultrafiltration and ion exchange chromatography to obtain the scorpion protein fractions with different molecular weights (3-10 ku,>10-50 ku and>50 ku). The embryos of transgenic zebrafishes were cultured in the above 10,100 and 500 μg/ml scor-pion’s proteins. Intersegmental vessels of the transgenic zebrafishes were counted under the fluorescence microscope to optimize the most suitable scorpion’s protein molecular weight. The vessels were counted again with >50 ku scorpion protein component 1 and 2,so as to select suitable component.RESULTS:The >50 ku scorpion’s protein fraction component 1 with the mass concentration of 500 μg/ml had the highest inhibitory activity for the angiogenesis of the transgenic zebrafish,with inhibitory rate of 92.59%. CONCLUSIONS:Scorpion’s protein and its fractions have the activity of angiogenesis inhibition,which may be one of anti-cancer mechanisms of scorpion.
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The four properties theory of traditional Chinese medicine (TCM) is an important basis for the guidance of TCM clinical medication.However,TCM modernization has produced a large number of active substances with unknown medicine properties.Therefore,it is urgent to build a new drug development system guided with TCM four properties theory.From the view of that four properties of TCM must have their material basis,and based on the known knowledge on TCM four properties,this paper proposed strategies of new drug research by cloning TCM partial components based on theTCM model of standard level property. A certain number of representative standard level property TCM drugs were selected to conduct system separation for the establishment of aTCM model of standard level property with the proportion and content of each separated component as a reference.The standard model was used as a boundary.Contents and proportions of the experimental TCM separation components deviated from thisstandard model were classified as hot or cold properties.From cloning of the partial components of TCM with hot or cold property medicine,the quality standards of partial components of experimental TCM were also studied,in order to investigate their corrective actions and mechanisms.The TCM partial components were expected to form a new TCM drug with the four properties theory of TCM as guidance.
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Objective To investigate the optimum extraction of total flavonoids and baicalin in baical skullcap root. Methods Ultraviolet spectrophotometry was used to detect the content of total flavonoids, HPLC was used to detect the content of baicalin. With the test of extracting method, time, frequency, and adding water volume, the optimum extracting craft was determined. Results The best craft for extraction was:heating water for recirculation, setting extracting time for 1.5 h, extracting frequency for 3 times, and water volume for 8 times of baical skullcap root. Conclusion The optimized process for extraction are stable, reasonable and feasible.
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Objective To optimize the extraction technology of ferulic acid (FA) in Ligusticum chuanxiong Hort. and tetrahydropalmatine in Corydalis yanhusuo from Kexintong dropping pill. Methods The extraction technology of L. chuanxiong was screened by the orthogonal test. The extration was refined with the high aperture jaffaite. The technology parameter for extracting C. yanhusuo by the classic method of acidic dissolution and alkaline precipitation was optimized by the orthogonal test. Results The more effective way to extract FA from L. chuanxiong was to extract with 12 times amount of 75% alcohol under reflux for 3.5 h. The chuanxiong extract of pH 3 at a flow rate of 5 ml/min on DM-130 macroporous resin column, was eluted with 80% ethanol. The eluent was collected and dried under vacuum. The more effective way to extract tetrahydropalmatine in C. yanhusuo was extracted with 22 times amount of acidic water (pH 4)for 3 h. Conclusion The optimum extraction is reasonable and practicable with high extraction ratio.
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<p><b>OBJECTIVE</b>To research on the substantial foundation of the medical speciality of Chinese traditional medicines from immunogenicity.</p><p><b>METHOD</b>Control antigen with hot nature was prepared from the mixture of the aqueous extracts of three Chinese traditional medicines with three typical hot nature of Alpinia officinarum, Cinnamomum cassia and Curculigo orchioides, while that with cold nature prepared with Rheum palmatum, Anemarrhena asphodeloides, Coptis chinensis, and polyclonal antibody was prepared by immunizing rabbit with control antigen. Dot blotting was performed between the polyclonal antibody of control antigen and the aqueous extracts of nine Chinese traditional medicines on a piece of PVDF membrane, and the blotting signals were analyzed by the software of Quantity One.</p><p><b>RESULT</b>Blotting signals with hot control antigen of nine Chinese traditional medicines in descending were Zingiber officinale, Aconitum carmichaeli, Eucommia ulmoides, Fraxinus rhynchophylla, Lonicera japonica, Anemarrhena asphodeloides, Coptis chinensis, Rheum palmatum and Phellodendron chinense, which degree of similarity to control antigen in peak value were 57.33%, 43.56 %, 34.16%, 30.2%, 28.81%, 26.53%, 21.68%, 17.62% and 14.85%, respectively. Blotting signals with cold control antigen were Rheum palmatum, Anemarrhena asphodeloides, Coptis chinensis, Phellodendron chinense, Zingiber officinale, Lonicera japonica, Fraxinus rhynchophylla, Eucommia ulmoides and Aconitum carmichaeli in descending, of which degree of similarity to cold control antigen in peak value were 55.22%, 54.23%, 46.72%, 34.08%, 30.3%, 24.48%, 24.33%, 20.35% and 15.17%, respectively. Results of cluster analysis with Wistar's method showed that nine medicines were classified into two groups, one group included Phellodendron chinense, Anemarrhena asphodeloides, Coptis chinensis, Rheum palmatum, another was Zingiber officinale, Aconitum carmichaeli, Eucommia ulmoides, Fraxinus rhynchophylla, Lonicera japonica.</p><p><b>CONCLUSION</b>Blotting signals of nine medicines with control antigen regularly varied with the alteration of medicine nature. The more similarity degree of the tested medicine to control antigen was smaller, the more distance of the tested medicine to control antigen was further. Dot immunoblotting was a practical and effective new method in researching the substantial foundation of the medical speciality of Chinese traditional medicines.</p>
Subject(s)
Animals , Male , Rabbits , Antibodies, Monoclonal , Allergy and Immunology , Antibody Formation , Antigens , Allergy and Immunology , Blotting, Western , Cold Temperature , Drugs, Chinese Herbal , Chemistry , Classification , Hot Temperature , Immunoblotting , Methods , Medicine, Chinese Traditional , MethodsABSTRACT
Objective To optimize the prescription matrix proportion of Bitongning Patch and observe its percutaneous absorption in vitro. Methods U6(66) Uniform design was performed to investigate the influence of scopolamine and aconitine in the proportion for 24 h cumulative permeation in vitro. The drug permeation behavior through excised mouse skin was studied with improved Franz diffusion cell. Results With the matrix ratio for PVA: 11.02%, PVP: 29.93%, glyceroh 7.42%, azone: 4.95%, and propanediol: 11.06%, respectively, the Patch had the maximal cumulative permeation amount and cumulative permeation curve coincided with the Higuchi Equation. Conclusion The prescription composition optimized by uniform design meets the requirement of skeleton-controlled transdermal delivery system.
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AIM: To establish a quantitative analytical method of ferulic acid in Kexintong Dripping Pills (Moschus, Rhizoma Chuaxiong, Rhizoma Corydalis, etc.) based on the capillary zone electrophoresis. METHODS: P-Nitrobenzoic acid was adopted as the internal substauce with a 30 mmol?L -1 borax solution at equal electrophoresis current of 50 ?A, detection wavelength at 295 nm, uncoated fused silica capillary of 75 ?m?50 cm, temperature at 20 ℃. Sample was injected for 5 s at pressure of 20PSI. RESULTS: Good linearity was showed in the concentration range of 4～264 mg?L -1 , RSD was 1.71%(n=6), and the recovery was 100.47%?2.1%. CONCLUSION: The result indicates the proposed method is convenient, rapid, accurate and reliable, it is a proper method for the quantitative analysis of ferulic acid in Kexintong Dripping Pills.
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AIM: To prepare Bitongning Patch(Flos Daturae, Moschus and Camphora, etc.) and study its in vitro release characteristics. METHODS: Bitongning Patch was prepared with high polymer hydrophilic material as matrix. The determination method of scopolamine was established by HPCE. The in vitro release was studied according to the requirement of Chinese Pharmacopoeia(2000) and the percutaneous absorption of Bitongning Patch was evaluated in use of the Franz diffusion cell. RESULTS: The release mechanism of Bitongning Patch in vitro coincided with Higuchi Equation. The release rate was 0.4698mg?cm -2?h -1/2. Scopolamine in Patch permeated in vitro at the speed of 0.067mg?cm -2?h -1 by zero-order kinetics. CONCLUSIONS: Bitongning Patch is a skeleton controlled TDDS whose permeation speed is limited by skin.
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OBJECTIVE:To study pharmacodynamics of different approaches of using low peptide of E’ jiao. METHODS: Anemia model rabbits and immunosuppressed model mice were induced by injecting with phenylhydrazine hydrochloride subcutaneously and hydrocortisone via i.p. respectively then treated with low peptide of E’ jiao (1～3 kD) via i.p., low peptide of E’ jiao of lower than 5 k and 5% E’ jiao solution via i.g. The values of Hb, RBC, WBC in model rabbits were measured after 7 days of medication and spleen weigh index was detected after 6 days of medication. The healthy mice were treated with low peptide of E’ jiao (1～3 kD) via i.p., low peptide of E’ jiao of lower than 5 k via i.g. and glucose solution via i.g. Then the load swimming time was determined after 14 days of medication. RESULTS: Chemical medicine-induced anemia was improved by 1～3 kD low peptide of E’ jiao increasing the amount of Hb. Low peptide of E’ jiao of lower than 5 k could not produce such an good effect on anemia but was eaqul to oral E’ jian in increasing the value of RBC, WBC. Low peptide of E’ jiao was helpful to or had effect on anti-fatigue and remarkably increased spleen index of mice. CONCLUSION: Low peptide of E’ jiao used by different approaches can strengthen immunity.