Brucea javanica
1、       This product is the dried and mature fruit of Brucea javanica (L.) merr., a bitter wood plant. The fruits are harvested when they are ripe in autumn, removed impurities, and dried in the sun.
      [properties] this product is oval, 6 ~ 10mm long, 4 ~ 7mm in diameter. The surface is black or brown, with raised reticular wrinkles, the mesh is irregular polygonal, there are obvious edges on both sides, the top is tapered, and there are concave fruit stem marks at the base. The fruit shell is hard and brittle, and the seed is oval, 5-6mm long and 3-5mm in diameter, with white or yellowish white surface and reticulation; The seed coat is thin, and the cotyledons are milky white and oily. The breath is slight and the taste is extremely bitter.
      [identification] (1) the peel of this product is brown. Epidermal cells are polygonal and contain brown matter. The parenchymal cells are polygonal, containing calcium oxalate cluster crystals and square crystals, and the diameter of cluster crystals is about 30 μ M. Stone cells are round or polygonal, with a diameter of 14-38 μ M. The seed powder is yellowish white. . The endosperm and cotyledon cells were obscurely powdered.
      (2) Take 1g of crude powder of the product, add 20ml of petroleum ether (60 ～ 90 ℃), sonicate for 30 minutes, filter, and add 20ml of petroleum ether (60 ～ 90 ℃) to the filtrate as the test solution. Another 1g of Brucea Javanica reference medicinal material was prepared into the reference medicinal material solution by the same method. Then take oleic acid reference substance and add petroleum ether (60 ～ 90 ℃) to make a solution containing 3mg per 1ml as the reference solution. According to the test of thin-layer chromatography (general rule 0502), suck 1 ~ 3 μ l of each of the above three solutions, respectively dot on the same silica gel G thin-layer plate, use petroleum ether (60 ~ 90 ℃) - ethyl acetate glacial acetic acid (8.5:1.5:0.1) as the developing agent, develop, take out, dry, and smoke in iodine vapor until the spots are clear. In the chromatogram of the test sample, spots with the same color appear at the corresponding positions of the chromatogram of the control medicinal material and the chromatogram of the control sample.
      [inspection] impurities shall not exceed 2.5% (general rule 2301).
      .
      The total ash content shall not exceed 6.5% (general rule 2302).
      [content determination] determine according to gas chromatography (general rule 0521).
      Chromatographic conditions and system suitability test polyethylene glycol 20000 (PEG-20M) capillary column (column length: 30m, inner diameter: 0.25mm, membrane thickness: 0.25 μ m); The detector temperature is 250 ℃ (FID); The temperature of the injection port was 250 ℃; The column temperature was 205 ℃; The split ratio was 20:1. The number of theoretical plates shall not be less than 5000 according to the peak of oleic acid.
      Determination of correction factor take an appropriate amount of oleic acid reference substance, weigh accurately, and add n-hexane to make a solution containing 3mg per 1ml. Accurately measure 5ml, put it into a 10ml plugged test tube, blow dry with nitrogen, add 2ml of 0.5mol/l potassium hydroxide methanol solution, put it in a 60 ℃ water bath for Saponification for 25 minutes, until all the oil droplets disappear, cool it, add 2ml of 15% boron trifluoride ether solution, put it in a 60 ℃ water bath for methyl esterification for 2 minutes, cool it; Precisely add 2ml of n-hexane, shake, add 1ml of saturated sodium chloride solution, shake, stand still, and take the upper solution as the reference solution. . Accurately measure 1ml of the reference solution and 1ml of the internal standard solution, shake well, suck 1 μ L, inject it into the gas chromatograph, determine and calculate the correction factor.
      . Accurately measure 3ml, and prepare the test solution in the same way as the reference solution preparation method from "place in 10ml plugged test tube and blow dry with nitrogen". Precisely measure 1ml of the test solution and 1ml of the internal standard solution, shake well, suck 1 μ L, inject into the gas chromatograph, and determine.
      According to the calculation of dry product, the content of oleic acid (c18h34o2) shall not be less than 8.0%.
      Decoction pieces
      [processing] remove the shell and impurities.
      [nature, taste and meridian tropism] bitter, cold; There is little poison. .
      [functions and indications] clearing away heat and toxin, intercepting malaria and stopping dysentery; . For dysentery, malaria; External treatment of warts, corns.
      [usage and dosage] 0.5 ~ 2G, wrap it with longan meat or swallow it in capsules. .
      [storage] put it in a dry place.


2、 Chemical constituents of Brucea javanica
Brucea Javanica contains more than 30 bitter components with anticancer activity that are structurally similar to quassin: Bruceine aBrucea javanicaB、Brucea javanicaC、Brucea javanicaD、Brucea javanicaE、Brucea javanicaF、Brucea javanicaG、Brucea javanicaH、Brucea javanicaI，Brucea Javanica bitter alcohol（brusatol）， Dehydrobrusatol, dehydrobruceantinol, dehydrobruceine A and B, dihydrobruceine,Crow gall Pavilion（bruceantin）， ,Brucea Javanica glycoside a（yadanzioside）、Brucea Javanica glycosideB、C、D、E、F、G、H、I、J、K、L、M、N、O、p， Bruceoside A and B, javanicin, etc. It also contains cauliflower lignan a, 4-ethoxycarbonyl-2-quinolone,Vanillic acid（vanillic acid），Hyperin（Hyperoside）， Luteolin 7-o - β - d-glu-coside,Carotene（daucosterol）。 It also contains oil (Brucea javanica oil), oleic acid with tumor suppressive activity and inactive components: tcmlibiolein, etc.

3、 Pharmacological effects of Brucea javanica
1. antiparasitic effect: 1.1. anti amoeba activity and cytotoxic effect of Brucea Javanica: the activities of Brucea Javanica butanol extract, jaydanin and Brucea Javanica bitter C against dysentery amoeba (entamoebahistolitica) and the cytotoxic effect on guinea pig ear keratinocytes (GPK) were determined in vitro.
1.2. both degreased Brucea Javanica water extract and ether extract can kill amoeba infected feces. .
1.3. antimalarial effect: 1.3.1. in vitro antimalarial effect of crude extract of Brucea Javanica and its active ingredients on drug-resistant strains of Plasmodium falciparum: morphological method was used to determine the in vitro antimalarial activity of crude extract of Brucea Javanica with four solvents against Plasmodium falciparum, and the results showed that chloroform extract had the strongest activity. Three compounds, Brucea Javanica a, Brucea Javanica B hydrate and Brucea Javanica C, were isolated and purified from the chloroform extract of Brucea Javanica. The in vitro antimalarial activities of these three chemical components against naturally isolated Plasmodium falciparum strains were determined. The results showed that Brucea Javanica A and Brucea Javanica B hydrate had similar antimalarial activities (ID50 of 8.66 and 8.15ng/ml), while Brucea Javanica C had higher antimalarial activity (ID50 of 1.95ng/ml), which was significantly higher than the new antimalarial drug mefloquine (ID50 of 6.26ng/ml)difference (p< 0.041).
1.3.2. effect of Brucea Javanica lignin on chloroquine resistant Plasmodium falciparum strains: the activity of a series of Brucea Javanica lignin components against chloroquine resistant Plasmodium falciparum strains in vitro was determined, including 9 components ic50< 0.05μg /ml， The in vitro antimalarial activity of yadanzioside F and I and its glycosyl Brucea Javanica bitter B were much less than that of yadanzioside F and I. The inhibitory effects of different solvent extracts of Brucea Javanica and some components of Brucea javanica on Plasmodium parasitemia in mice were also determined. .
1.3.3. the in vitro antimalarial activities of buceoside-a, Brucea Javanica and bruceolide against chloroquine resistant strains of Plasmodium falciparum showed that Brucea Javanica had high antimalarial activity (half effective amount = 7.58ng/ml), bruceoside a had no activity (half effective amount = 581.14ng/ml), and bruceoliole had minimal activity (half effective amount = 75.20ng/ml).
1.3.4. anti other parasitic effects: the crude extract of Brucea Javanica can drive away intestinal nematodes and stripworms in dogs. . A bitter sweetener extracted from Brucea Javanica can kill Paragonimus adults within 24 hours at a concentration of 1:10000 in vitro. It can kill Trichomonas, Paramecium and urozoa. Contact of 5-10% cold extract with mosquito larvae and eggs for 18-48 hours can kill them all.
2. antitumor effect: 2.1.10% Brucea Javanica Oil intravenous emulsion antitumor effect: 2.1.1. anticancer effect in mice: 2.1.1.1. effect on Ehrlich ascites cancer: mice weighing 20 ± 0.5g, 10 mice in each group, administered the next day after inoculation with Ehrlich ascites cancer. . The results showed that the average survival days of the two dose groups were significantly longer than those of the control group (P < 0.001).
2.1.1.2. effect on solid tumors S37 and S180: mice weighing 20 ± 0.5g were subcutaneously inoculated with cancer cell suspension in the axilla of the right forelimb, with 10 mice in each group. From the next day after inoculation, the tumor was locally administered with 2.5g/ (kg. day) × 7 days. The animals were sacrificed 15 days after administration, and the body weight, tumor weight and tumor inhibition rate were recorded. The results showed that Brucea Javanica Oil intravenous emulsion had an anticancer effect on mouse solid tumors S37 and S180.
. The results of morphological method showed that Brucea javanica oil emulsion had direct damage effect on Ehrlich ascites cancer cells, liver cancer ascites cancer cells, such as membrane degeneration, membrane rupture and nuclear pyknosis. The Mouse Spermatogonia method proved that Brucea javanica oil emulsion could inhibit the mitosis of Mouse Spermatogonia.
2.2. killing kinetic effect of Brucea javanica oil emulsion on Ehrlich ascites cancer cells: the effect of 10% Brucea javanica oil emulsion on S phase, G2 phase and go phase cells of Ehrlich ascites cancer mice inoculated for 7 days was observed by autoradiography. The results of S-phase killing showed that 8 hours after administration, the damage of S-phase and non-s-phase cells showed a peak, but the damage percentage of S-phase cells was higher than that of non-s-phase cells. . The experiment of 3H TDR labeling and colchicine blocking showed that the increase of labeling division index (PLM) in the administration group was significantly lower than that in the control group. Because no damaged M-phase cells were found during drug treatment, and cells in various stages of mitosis could be seen, the no obvious rise in MI after drug treatment was not caused by the direct damage of M-phase cells by drugs, indicating that Brucea javanica oil emulsion can inhibit or damage G2 phase cells, thus reducing and prolonging the entry of G2 phase cells into M phase. According to the continuous labeling method, 47.8% of go phase cells were identified, and the damage rate of go phase cells was 17% 5 hours after administration, indicating that Brucea javanica oil emulsion also has a certain killing effect on go phase cells. Autoradiography confirmed that Brucea javanica oil emulsion had obvious inhibitory effect on the incorporation of 3H TDR into DNA biosynthesis. The Li was 11% at 0.5 h and 33% at 24 h after administration, indicating that the drug had reversible inhibition on Ehrlich ascites cancer cells.
3. anti leukemia effect: Brucea javanica, which contains the bitter lignin glycosides brnceosidea, B, yadanziosides A-H, yadanziosides F, I, J, l, P, shows ANTI LEUKEMIA activity. Brucea Javanica bitter alcohol showed effective anti leukemia effect on P-388 leukemia (0.125mg/kg/ day in mice, t/c=158). Cleomiscosina had activity on P-388 lymphoid leukemia in vitro (half effective dose = 0.4 μ g/ml).
4. anti other tumor effects: skin cancer and papilloma induced by methyl cholanthracetone in mice were tested, and it was proved that Brucea Javanica paste and water agent can cause tumor cells to undergo degeneration and necrosis when applied locally. But it also has a similar effect on normal tissues. .