CN1947793A - Application of Entamoeba histolytica for treating malignant noumenal tumor - Google Patents
Application of Entamoeba histolytica for treating malignant noumenal tumor Download PDFInfo
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- CN1947793A CN1947793A CNA2005100218619A CN200510021861A CN1947793A CN 1947793 A CN1947793 A CN 1947793A CN A2005100218619 A CNA2005100218619 A CN A2005100218619A CN 200510021861 A CN200510021861 A CN 200510021861A CN 1947793 A CN1947793 A CN 1947793A
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- tumor
- ameba
- histolytica
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- solid tumor
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Abstract
An application of the toxic histolytic ameba in treating the malignant solid tumor is disclosed, which features that said toxic histolytic ameba is used as the biologic knife to excise malignant solid tumor.
Description
(1) technical field:
The present invention relates to the amoebic application of histolytica's class, the application of histolytica's class ameba in the treatment malignant solid tumor specifically.
(2) background technology:
Tumor is one of common disease that threatens human health, is only second to cardiovascular disease, and its morbidity is the trend that rises year by year.At present, the more employing of the clinical treatment of tumor is based on the Comprehensive Treatment of operation, comprises chemotherapy, physiotherapy (radiation, ultrasonic, thermotherapy, cryogenic technique etc.), Biotherapeutics, traditional Chinese medical herbal treatment, but all undesirable.The operative treatment cancer patient damages big, easily shifts, and normal structure of put, chemotherapy easily being killed and wounded patient and destruction patient immune system patient's resistance is reduced, and patient's medical expense height, weak curative effect, side effect are big.Therefore, the Biotherapeutics research field of the new malignant tumor of developing is the important subject of current medical circle.
Amoeba histolytica (Entamoeba histolytica) is the parasite among a kind of human colon of colonizing in, and belongs to the Lobosea (Class lobasea) of meat foot flagellum door (Phylum Sarcomastigophora).At present as yet not relevant for the report of using histolytica's class ameba treatment malignant solid tumor.
(3) summary of the invention:
The objective of the invention is amoeba histolytica is applied to treat malignant solid tumor.
Technical scheme of the present invention is as follows: a kind of purposes of amoeba histolytica, it is to be used for the treatment of malignant solid tumor.Be about to amoeba histolytica's trophozoite of tool virulence, inject the tumor center position behind location such as B ultrasonic, CT, nuclear magnetic scanning, as biological cutter localized resection malignant solid tumor, with the treatment malignant solid tumor.Its dosage is each tumor center position disposable injection amoeba trophozoite 5-10ml.
The application of histolytica's class ameba in the treatment malignant solid tumor is that histolytica's class ameba is made inoculated tube (2-20ml/ pipe).
Show through human body or zoopery result, the worm strain (Entamoeba histolytica Schaudinn) that causes the invasive amebiasis in the amoeba histolytica (Entamoeba histolytica) is adsorbable, dissolve, engulf tissue, equally also can adsorb, dissolve, engulf the malignant entity tumor tissue, the factor that plays a role is:
1. the malignant solid tumor histiocyte is adsorbed in galactose/acetylamino galactosamine coagulation 260kDa-mediation.
2. ameba perforin (amoeba pores)-form poroid destruction at the malignant solid tumor histiocyte makes permeability of cell membrane change cellular atrophy.
3. cysteine proteinase-dissolving malignant solid tumor histiocyte.
4. ameba has ingratiating with property of chemistry to host's neutrophilic granulocyte.Granulocyte is touched to go out to disengage poisonous oxide after molten and aggravate the malignant solid tumor disorganization by trophozoite.
5. the lectins of trophozoite pellicle (lectin) makes the coagulation of malignant solid tumor tissue blood, hemostasis, thrombosis; The tumor tissue blood supply reduces on the one hand, and the tumor tissue cell is hungry dead.The tumor tissue cell is restricted by the blood diffusion on the other hand.
6. can excite body fluid and cellullar immunologic response behind the routine host infection histolytica class ameba.Malignant solid tumor tissue lymph cell aggregation, secrete various IgG, IgM, IgA and IgE antibody and contain cytokines such as enriching gamma interferon, TNF, IL, and then activated macrophage and neutrophilic granulocyte, make the enhancing phagocytic power, produce superoxides, kill and wound tumor tissue, and around the solid tumor tissue, form an immune ring of encirclement, make the tumor tissue cell directly soak into diffusion and be restricted.
Zoopery is the result also show, histolytica's class ameba makes malignant solid tumor become ameba pus infections through above mechanism, after having finished biology excision to malignant solid tumor, take the Therapeutic Method (drain of various amebic abscesses again, part and the treatment of whole body anti-amebic thing etc.) cure amoeba histolytica's pus infections, promptly reach at last and use the purpose of histolytica's class ameba as a kind of biological cutter localized resection malignant solid tumor.Therefore, use histolytica's class ameba treatment malignant solid tumor and not only solved the damage of operative treatment cancer patient greatly, easily shift, put, chemotherapy infringement patient's normal structure makes reduction of patient's resistance and the unfavorable major defect of other biological therapy effect with immune system, and medical expense is low, good effect, side effect is low, has opened up the new way of treatment of cancer.
The invention will be further described by the following examples.
(4) specific embodiment:
The biological excision treatment of the foundation of embodiment 1 nude mice people hepatoma Metastasis model LCI-D20 (LCI-D35) and ameba
(1) the amoebic cultivation of pathogenic histolytica's class
The main Robinson that uses " the s culture medium, not only can cultivate histolytica's class ameba, also can cultivate multiple amebas such as breathing out Men Shi entamoeba, entamoeba coli, amoeba limax.General application 6-7ml or littler spiral cover culture tube.
1.1 constituent:
(1) saline agar slant: dissolving 15g agar powder and 7-8gization sodium are divided in the sterilization of small test tube mesohigh (121 ℃ 15min), make it form the inclined-plane when agar is cooled to incline about 75 ℃ to putting in the 1000ml distilled water.
(2) erythromycin: the 0.5g laboratory is placed sterile chamber with the erythromycin powder, add the 20ml70% dissolve with ethanol, place more than the 2h, then add aquesterilisa for 4 ℃ to 50ml.
(3) rice flour: the Semen oryzae sativae powder is through autoclave sterilization or 180 ℃ of dry sterilizations.
(4) preparation 50mol Potassium Hydrogen Phthalate, PH6.3, autoclaving.
(5) serum: 56 ℃, the 30min deactivation, even the cattle of not deactivation or horse serum all can use.
(6) R solution stock solution: NaCL 50g, (NH
4)
2SO
410g, citric acid .2H
2O 20g, MgSO
47H
2O 0.5g, KH
2PO
45g, lactic acid (90% purity) 4ml add water to 950ml, regulate PH to 7.0, and final pondage is to 1000ml, and the packing autoclaving is prepared into the storage working solution.During use the 100ml stock solution is added the 850ml distilled water, regulate PH7.0 packing autoclaving.
(7) BR solution: 25ml R working solution and 1 clone's escherichia coli, 37 ℃ of joltings were cultivated 24-48 hour.
(8) BRS solution: in above-mentioned BR solution, add equivalent serum, continue cultivation and got final product in 24-48 hour.
1.2, operating procedure:
In containing the culture test tube of agar slant, add 10mg rice flour, 120 μ l erythromycin liquid and Potassium Hydrogen Phthalate and 4: 1 mixed liquors of BRS liquid of enough hiding the inclined-plane amount, add a small amount of (about 50mg) feces, mixing, cultivate after 24 hours for 37 ℃, remove culture supernatant, add and go into 4: 1 mixed liquors in right amount and add 60 μ l erythromycin and rice flour.37 ℃ are continued to cultivate after little 48 o'clock, get one in rice flour and feces mixture, have or not trophozoite with iodine staining or direct observation; If do not find that polypide adds rice flour again, cultivate observation in 24 hours again.The nymph body positive can change continuation cultivation in the fresh culture over to cultivating mixed liquor on a small quantity, is transferred species.Get and cultivate back 72h, about 3.0 * 10 polypides of amoeba histolytica/10ml places tool plug 15ml pipe, and the centrifugal 5min of 120g abandons supernatant, washes 3-5 time repeatedly with physiological saline solution, is made into 10 * 10 with physiological saline solution
6Polypide/10ml/ pipe, 0-4 ℃ of transportation put in capping, incubates with 37 ℃ of water-bath temperature before the inoculation and can inoculate use after 30 minutes.
(2) the biological excision treatment of the foundation of people's hepatoma Metastasis model LCI-D20 (LCI-D35) and ameba.
2.1 the HCCLM3 cancerous cell HCCLM3 (cell strain is provided by Fudan University's liver cancer research) of logarithmic (log) phase growth.Culture fluid abandons, and uses phosphate buffer (PBS) washed cell 1 time, uses trypsinization, and eluting from culture bottle is made single cell suspension.Adjust concentration to every 0.2ml 5 * 10
6Individual/cell, it is subcutaneous that the right omoplate of nude mouse (being provided by laboratory animal portion of Shanghai medicine institute of the Chinese Academy of Sciences) is provided.When treating tumor length, put to death nude mice, get tumor tissues, remove surface fiber, blood vessel to about 1cm.Put into the culture dish of DMEM culture fluid after washing with PBS liquid, be cut into the 1.0mm3 piece of tissue.Sulphuric acid pentobarbital sodium intraperitoneal injection of anesthesia nude mouse (40mg/kg) with 3%.Get the about 1.5cm of left subcostal incision, the layering open abdomen enters the abdominal cavity, exposes leftlobe of liver.Do the 0.3cm otch on its surface, hemostasis by compression is implanted the tumor tissue piece in the liver, and the figure-of-eight suture liver is sent liver back to abdominal cavity, and holostrome is sewed up stomach wall.
2.2 animal grouping: all animals behind in-situ inoculating tumor piece the 10th day, former operative scar of fusiform excision and otch surrounding skin, subcutaneous tissue, muscle, carefully expose transplanted tumor after advancing abdomen, the mice of transplanted tumor success is divided into two groups, every group 10, amoeba histolytica's Biotherapeutics group is with the Entamoeba histolytica trophozoite 10 of tool virulence
5～10
6/ ml 0.1ml/ only injects liver inoculation tumor growth site center, and matched group injects 0.1ml/ does not only have amoebic ameba culture fluid, 7 days formation liver ameba pus infectionss (available B ultrasonic is observed).When growing to one times big of liver inoculation tumor body, liver ameba pus infections comes into effect the treatment of anti-amebic thing, lumbar injection levofloxacin 250mg/kg, and/d, successive administration 3W,
B ultrasonic is observed liver ameba pus infections healing state and is determined treatment time, the matched group intraperitoneal injection of saline.Observe the relatively life span of two treated animals, when experiment finishes, get that hepatoncus position tissue is weighed, pathological section and comparative control statistics T check analysis.The results are shown in Table 1
Table 1 amoeba histolytica directional inoculation is to the influence of nude mice people hepatocarcinoma (LCI-D35) metastasis model
| Grouping | Number of animals (only) | LCI-D35 cell 6 * 106/ml/ only | Mean survival time x ± s (my god) | Heavy x ± the s (mg) of liver tumor piece | Pathological section microscopy result |
| Normal saline matched group ameba treatment group | 20 20 | 0.1 0.1 | 20.3±4.5 41.4±6.2 * | 12.3±2.2 2.4±1.6 * | Meet the performance of hepatocarcinoma pathological changes and meet the performance of liver abscess later stage |
● compare P＜0.01 with the normal saline group
Embodiment 2. uses the clinical trial of amoeba histolytica as biological cutter localized resection adversity of the liver solid tumor
The constitutional limitation liver cancer patient of clinical definite is behind location such as B ultrasonic, CT, nuclear magnetic scanning, with the Entamoeba histolytica trophozoite 10 of tool virulence
5～10
6/ ml 5-10ml/ injects the liver neoplasm centre, when behind 2-3W, forming hepatocarcinoma position liver ameba pus infections (available B ultrasonic is observed) and treating that liver ameba pus infections scope surpasses former liver neoplasm 1CM, liver cancer marker (AFP) content is just often in the blood after testing, can think that liver tumor excised by biology, begin by local drain and local injection anti-amebic thing, cooperate the anti-ameba of whole body, antibacterial therapy and other support trophotherapy simultaneously, cure the ameba liver abscess, reach malignant tumor purpose clinical or that cure fully at last.
Embodiment 3. uses the clinical trial of histolytica's class ameba as biological cutter localized resection adversity of the lung solid tumor
The constitutional limitation patients with lung cancer of clinical definite is behind location such as B ultrasonic, CT, nuclear magnetic scanning, with the Entamoeba histolytica trophozoite 10 of tool virulence
5～10
6/ ml 5-10ml/ injects the lung tumor centre, when behind 2-3W, forming pulmonary carcinoma position lung ameba pus infections (available X-ray is observed) and treating that lung ameba pus infections scope surpasses former liver neoplasm 1CM (available X-ray, CT, nuclear-magnetism are checked), can think that liver tumor excised by biology, begin by local drain and local injection anti-amebic thing, cooperate the anti-ameba of whole body, antibacterial therapy and other support trophotherapy simultaneously, cure the ameba lung abscess, reach malignant tumor purpose clinical or that cure fully at last.
Embodiment 4. histolytica's class amebas as biological cutter localized resection can also be applied to breast carcinoma, intestinal cancer, malignant brain tumor, etc. the treatment of focal malignant tumor, with embodiment 2,3 roughly the same.
Claims (1)
1, the application of histolytica's class ameba in the treatment malignant solid tumor.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CNA2005100218619A CN1947793A (en) | 2005-10-10 | 2005-10-10 | Application of Entamoeba histolytica for treating malignant noumenal tumor |
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| CNA2005100218619A CN1947793A (en) | 2005-10-10 | 2005-10-10 | Application of Entamoeba histolytica for treating malignant noumenal tumor |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102697816A (en) * | 2012-06-21 | 2012-10-03 | 桂林医学院 | Application of escherichia coli in preparing medicine for treating hepatic malignant solid tumor by matching with entamoeba histolytica |
-
2005
- 2005-10-10 CN CNA2005100218619A patent/CN1947793A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102697816A (en) * | 2012-06-21 | 2012-10-03 | 桂林医学院 | Application of escherichia coli in preparing medicine for treating hepatic malignant solid tumor by matching with entamoeba histolytica |
| CN103301163A (en) * | 2012-06-21 | 2013-09-18 | 桂林医学院 | Application of escherichia coli mixed entamoeba histolytica in preparation of medicine for treating liver cancer solid tumor |
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