| With high death rate, Malignancy threaten mankind's healthiness badly. Because of side-effects such as immunity-restraining, marrow- restraining etc., it is hard to improve survival quality of cancer-patients by Chemotherapeutics medicine only. In recent years, study of TCM on anti-tumor is very prolific, with the deepening study of Molecule-biology, abecedarian mechanism-study has began to go on of TCM to anti-tumor on molecule level. Extraction of Hedyotis diffusa Willd. (Rubiaceous) have obvious anti-tumor activity, the clinical application enunciate that it can treat kinds of Malignancy, such as oesophagus-cancer, tummy-cancer, galactophore-cancer an so on. Having unknown of the matter-base and the mechanism, and it is pressed for clinical preparations, application of it have been limited highly. So it is requisite to study on the matter-base and the mechanism of Hedyotis diffusa Willd.Purposes:1. To summarize and analyse the international literatures of Hedyotis diffusa Willd.;2. By using indexes including amylase-content & flavone-content to evaluate quality of the medicinal materials in different provenances abecedarianly, and to offer theory - gist for standard-constituting;3. To extract , separate and purificate chemical constituents from Hedyotis diffusa Willd, by ethanol, in order to find potential anti-tumor compound;4. To purificate amylase of it which extracted by water, and to study the approachs of extraction and purification;5. To separate and purify chemical constituents from Hedyotis diffusa Willd. injector by ethanol, in order to find potential anti-tumor compound or chemical-mixture;6. To evaluate the ability of killing tumor-cell of the compounds or chemical-mixture abecedarianly, which were got from Hedyotis diffusa Willd. and it's clinical preparations; To evaluate the ability to control tumor of amylase abecedarianly, in body of little white- rats who have sickened for cancer. On base of all the work above, we want to find the potential matter-base and the mechanism, to answer why it can treat cancer, and to offer theory-gist for the manufacture of new, effectual, steady and safe clinical preparations.Methods:1. Using computer or handworked searches, by full using of CA, BP, CNKI, CMCI, WEB and so on, to get plentiful and all-sided literatures of Hedyotis diffusa Willd., in order to analyse it's study- survey;2. Using UV-colorimetry to establish simple and convenient methods to determine amylase-content & flavone-content, so we can evaluate quality of the medicinal materials in different provenances synthetically;3. Using solvent-extraction, chromatogram to extract, separate and purify chemical constituents from Hedyotis diffusa Willd. by ethanol; Using mp., spectrum-IR, NMR, MS-to conclude possible frame of chemical compounds which were got from Hedyotis diffusa Willd. ;4. Using orthogonal design, with amylase-content as the index, to find the best approachs of extraction for amylase extraction and purification;5. According to GB, to produce good-quality Hedyotis diffusa Willd. Injector, to find potential anti-tumor compound or chemical-mixture by chromatogram;6. To evaluate the ability for anti-tumor of amylase abecedarianly, in body of little white- rat who had got cancer; Using MTT-way to evaluate the ability to killing tumor-cell of the compounds or chemical-mixture abecedarianly, which were got from Hedyotis diffusa Willd. and it's clinical preparations.Results:1. Literatures summarization:Applying tools above, we collected about 90 literatures at all, among them, domestic literatures are focusing on study of quality, pharmacology, clinic; overseas literatures are focusing on study of Chemical Constituents. But there are no literatures about the matter-base and the mechanism of Hedyotis diffusa Willd. and it's clinical preparations; The preparations study is behindhand comparatively, such as Hedyotis diffusa Willd. injector-easy approachs, side-effects and unknown of matter-base.2. Study on quality of medicinal materials:2.1 Naphtha: Oil rate of medicinal materials are accordant to each other from Guangdong, Guangxi and Jiangxi Province, about 0.25 ~ 0.30%,primary compound is Hexadecanoic acid, and there is tiny difference in oil composing;2.2 Amylase: Using vitriol-phenol way, we established simple and convenient methods of determine amylase-content, standard curve got is Conc.=0.10493mg* Abs (r=0.99981) ; amylase-content is accordant to each other of three block of medicinal materials, about 11%, which from Jiangxi Province is most high;2.3 Flavone: Using Al3+-colorimetry way, we established simple and convenient methods of determine flavone-content, standard curve got is Conc.= 1.94900mg* Abs (r=0.99985) ; flavone-content of medicinal materials from Guangdong Province is clear higher than the other.Synthesizing three indexes above, we can say that quality of medicinal materials from Guangdong Province is the best in our experimentation.3. Study on extraction by ethanol:Consult to literatures, we separated and purified three compounds belonging to steroid or terpenoids, one of them is 8-sitosterol, the other is unknown for the moment.4. Study on approaches of extraction for amylase:4.1 Approach of extraction by water: By orthogonal design, we can ascertain the best approach is to extract medicinal material tow times: first time ,we should use lOtimes water, marinate 30min, boil it 2.5h, filtrate; then we should add 8times water, boil it 2.5h,filtrate;4.2 Approach of deposit by ethanol: By orthogonal design, we can ascertain the best approach is to condense extraction to fluidextract, then add and mill ethanol to degree of ethanol's concentration get to 80%, deposited 6h;4.3 Approach of eliminating protein: Experiment show that Tea-way is more compatible than Sev-way for Hedyotis diffusa Willd. to eliminating protein; By orthogonal design, we can ascertain the best approach is to eliminating protein one time ,with one time precipitator, deposited 2h.5. Study on chemical-mixture extracted from Hedyotis diffusa Willd. Injector:5.1 According to GB, we produced two block of Hedyotis diffusa Willd. injector, and their qualities accord with the State standard;5.2 By polyamide-silica gel pole chromatography, we get two chemical-mixtures: alkaloid and flavone; By TLC we know that there are least 5 compounds belonging to alkaloid.6. Study on pharmacology:6.1 Anti-tumor in body: In rat body, the amylase have the capability to treat transplanted S-180 caruncle, control rate is almost 40%, and there is plus dose-effect relation, especially the high-dose group behave marked difference (P<0.001); the amylase can also markedly increase the thymus and spleen indexes. Compared with Cy, though the control rate of Cy is more high, but there also more high side-effects when treated by Cy;6.2 The ability to kill tumor-cell: By MTT-way, we put up study on the ability tokill tumor-cell of the compounds or chemical-mixture abecedarianly, which were got from Hedyotis diffusa Willd. and it's clinical preparations. Results tell us that liver cancer-cell(SMMC-7721) is more sensitive to J-3, and colon cancer-cell(Lovo) is more sensitive to J-2, the control rates are all about 40%.Conclusion:Our experiments prove that compound or chemical-mixture, which were got from Hedyotis diffusa Willd. and it's clinical preparations, have the ability to anti-tumor; The results can also offer theory-gist for the standard-constituting, for expounding the matter-base and the mechanism, also for manufacture of new, effectual, steady and safe clinical preparations. But there are some study haven't finished faultlessly, so it is requisite to study more deeply on the matter-base and the mechanism of Hedyotis diffusa Willd..
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