CN107260703A - A kind of internal magnetic guiding delivering method of salmonella - Google Patents
A kind of internal magnetic guiding delivering method of salmonella Download PDFInfo
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- CN107260703A CN107260703A CN201710301114.3A CN201710301114A CN107260703A CN 107260703 A CN107260703 A CN 107260703A CN 201710301114 A CN201710301114 A CN 201710301114A CN 107260703 A CN107260703 A CN 107260703A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0002—Galenical forms characterised by the drug release technique; Application systems commanded by energy
- A61K9/0009—Galenical forms characterised by the drug release technique; Application systems commanded by energy involving or responsive to electricity, magnetism or acoustic waves; Galenical aspects of sonophoresis, iontophoresis, electroporation or electroosmosis
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/46—Ingredients of undetermined constitution or reaction products thereof, e.g. skin, bone, milk, cotton fibre, eggshell, oxgall or plant extracts
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/5073—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals having two or more different coatings optionally including drug-containing subcoatings
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/5094—Microcapsules containing magnetic carrier material, e.g. ferrite for drug targeting
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
A kind of internal magnetic for the salmonella for guiding equipment combination to build based on magnetic corpusculum, magnetic micro-capsule and magnetic force guides delivering method, wherein the magnetic micro-capsule includes：Contain magnetic corpusculum in the mixing drop being mixed to get by the nutrient solution and hydrogel degradation solution of the salmonella containing magnetotactic, the magnetotactic salmonella；Phosphoglyceride droplet, wherein including the mixing drop；And the hydrogel shell of coated ferroferric oxide magnetic nano-particle, it is wrapped in outside the phosphoglyceride droplet.Based on the magnetic corpusculum and magnetic micro-capsule, the present invention guides equipment by Non-contact Magnetic, and the targeting that combination realizes salmonella is delivered.
Description
Technical field
The salmonella that equipment combination is built is guided based on magnetic corpusculum, magnetic micro-capsule and external magnetic force the present invention relates to a kind of
Internal magnetic guiding delivering method, specifically, is related to the magnetotactic salmonella containing magnetic corpusculum and loads magnetotactic salmonella
Magnetic micro-capsule, and equipment is guided by Non-contact Magnetic in vitro, a kind of salmonella that combination is realized targets throwing in vivo
The method sent, belongs to biomedical applications field.
Background technology
There is the problem of medicine can not be at accurate enough and enough arrival sufferer in traditional molecular medicine targeting carrying method,
Therefore the targeting efficiency and fragmentation effect of the antineoplaston factor are improved, it is necessary to rely on new targeting transport vehicle.Compared to
Other transport vehicles, salmonella has hypoxemia, thermophilic tumor tissues characteristic.By chemoattractant receptor and power flagellum, it can
Expand with sensing, mobile, aggregation and in solid tumor, but because salmonella self drive is weaker, and body vessel environment is multiple
It is miscellaneous, it is allowed to be difficult to reach target organ tumor tissues.
Magnetotactic bacteria is the general name for the bacterium that a class can be moved along magnetic direction, and it is most characterized in significantly in intracellular
Special prokaryotic device-magnetic corpusculum.Magnetic corpusculum be with outer membrane coating, nanoscale, intracellular into catenation Fe3O4Or
Fe3S4Magnetic-particle, is magnetotactic bacteria navigation [document 1] as compass.Kolinko I etc. utilize Gene transfer techniques
Synthesizing magnetic nano particle, i.e. magnetic corpusculum chain in photosynthetic red spirillum body, have started the artificial conjunction of magnetotactic bacteria
Into method [document 2].Further, Martel realizes magnetotactic bacteria in artificial blood vessel using nuclear magnetic resonance (MRI) gradient fields
Motion control [document 3].Canadian biomedical engineering research institute-nanometer robot laboratory attempt controls to carry medicine in vitro
The moving direction of magnetotactic bacteria, breaches the problem [document 4] that macromolecular drug is difficult to permeate into solid tumor Fa Yang areas.But more than
Research it is many based on external simulated test, it is difficult to the influence for overcoming internal complex environment to deliver bacterium, therefore one kind need to be researched and developed
The method that magnetotactic bacteria targets delivery in vivo.
Document 1：Alphandéry E.Applications of magnetosome synthesized by
magnetotactic bacteria in medicine.Front Bioeng Biotechnol.2014,2(5):1-6
Document 2：Kolinko I,Lohe A,Borg S,.Raschdorf O,Jogler C,Tu Q,Posfai M,
Tompa E,PLITZKO JM,Brachmann A,Wanner G,Muller R,Zhang Y,Schuler
D.Biosynthesis of magnetic nanostructures in a foreign organism by transfer
of bacterial magnetosome gene clusters.Nat Nanotechnol.2014,9(3):193-197
Document 3：Martel S,Felfoul O,Mathieu JB,Chanu A,Tamaz S,Mohammadi M,
Mankiewicz M,Tabatabaei N.MRI-based Medical Nanorobotic Platform for the
Control of Magnetic Nanoparticles and Flagellated Bacteria for Target
Interventions in Human Capillaries.Int J Rob Res.2009,28(9):1169-1182
Document 4：Taherkhani S,Mohammad M,Daoud J,Martel S,Tabrizian M.Covalent
binding of nanoliposomes to the surface of magnetotactic bacteria for the
synthesis of self-propelled therapeutic agents.ACS Nano.2014,8(5):5049-5060
The content of the invention
For technical problem present in prior art, contain magnetotactic salmonella it is an object of the invention to provide one kind
Magnetic micro-capsule, and using the magnetic micro-capsule in vitro magnetic guiding equipment effect under internal magnetotactic salmonella targeting deliver
Method.
The present invention is achieved through the following technical solutions.
A kind of magnetic micro-capsule, including：
In the mixing drop being mixed to get by the nutrient solution and degradation solution of the salmonella containing magnetotactic, the magnetotactic salmonella
Contain magnetic corpusculum；
Phosphoglyceride droplet, wherein including the mixing drop；And
Magnetic hydrogel shell, it is wrapped in outside the phosphoglyceride droplet.
Preferably, the degradation solution is Sodium Alginate Hydrogel Films degradation solution.
Preferably, the bacterial density of the nutrient solution of the salmonella containing magnetotactic in the mixing drop is 1 × 105Individual/milli
Rise；The concentration of the Sodium Alginate Hydrogel Films is 0.01 grams per milliliter.
Preferably, the hydrogel shell passes through the marine alga acid solution comprising ferroferric oxide magnetic nano-particles and chlorination
Calcium solution crosslinks reaction and formed.
Preferably, the magnetic nano-particle concentration in the marine alga acid solution comprising ferroferric oxide magnetic nano-particles
For 0.005 grams per milliliter.
Preferably, the magnetic corpusculum is to pass through electricity by plasmid pTps-ABG6 complexs and plasmid pTps-XYZ complexs
Method for punching is built.
Present invention also offers the manufacture method of the magnetic micro-capsule according to any one of above technical scheme, including with
Lower step：
The bacterium solution of magnetotactic salmonella is inoculated in lysogeny broth (LB) nutrient solution without antibiotic, vibration training
Support to mid log phase, thalline is collected by centrifugation, washed with the aseptic deionized water of precooling 2 times, the bacterium after washing is suspended from ice
The aseptic deionized water of precooling, with electroporation by pTps-ABG6, pTps-XYZ plasmid transfection to salmonella, builds magnetic small
Body gene, so as to realize the magnetotaxis of salmonella.
Using the soft lithography of semiconductor technology, microchannel chip is processed；
LB nutrient solutions and the Sodium Alginate Hydrogel Films degradation solution of 0.0162 grams per milliliter containing the attenuation salmonella are mixed
Close；
By syringe pump, by the mixed liquor, phosphoglyceride, the alginic acid comprising ferroferric oxide magnetic nano-particles are molten
Liquid and mineral oil are injected separately into the microchannel chip, form marine alga acid solution parcel phosphoglyceride droplet and include mixed liquor again
The structure of droplet；
Marine alga acid solution forms marine alga acid solution droplet by mineral oil liquid pinch off, and the microlayer model is injected into calcium chloride solution
In, alginic acid droplet will crosslink reaction with calcium chloride solution, hydrogel shell be formed, so as to form magnetic microcapsule structure.
Preferably, the depth of the fluid channel is 100 microns.
Delivering method is guided present invention also offers a kind of magnetic of salmonella, is comprised the following steps：
Step 1: manufacture includes the magnetic micro-capsule according to one of above technical scheme of the salmonella；
Step 2: the magnetic micro-capsule to be delivered to the first position near target location；
Step 3: guiding magnetic micro-capsule described in the introduction by magnetic field of equipment to reach the second near target location using magnetic force
Put；
Step 4: the magnetic micro-capsule ruptures in the second place, so as to discharge the salmonella；Afterwards, institute is made
Salmonella is stated under the guiding of magnetic field gradient, target location is moved to.
Preferably, the magnetic force guiding equipment includes mechanical arm and electromagnetism tweezer；It can adjust described by the mechanical arm
The tip location of electromagnetism tweezer, so as to form the magnetic field gradient.
Preferably, in the step 4, the magnetic micro-capsule containing the salmonella passes through its own internal package
Degradation solution ruptures its timing, so that the salmonella be discharged.
Beneficial effect：
The present invention wraps up the magnetic micro-capsule of magnetotactic salmonella by being built based on microflow control technique；It is finally based on this bag
The magnetic micro-capsule of salmonella is wrapped up in, the targeting Delivery method of the magnetic force guiding controlled based on mechanical arm is built, collection organ can be achieved
Interior intervention, big blood vessel micro-capsule magnetic conductance are delivered tends to integral active three-level delivery system certainly with thin vessels salmonella magnetic conductance,
High targeting, the delivery of high concentration of the treatment factor are broken through, the advanced colorectal cancer shifted at present with multiple organ can be overcome to control
Treatment method efficiency is low, it is specific not high the problem of, provide new approach for the therapeutic efficiency that improves targeted drug.
Brief description of the drawings
Fig. 1 is the method schematic diagram of the present invention
Fig. 2 is micro-channel structure schematic diagram
The detail view of local location 16 in Fig. 3
Reference：1. salmonella flagellum；2. salmonella；3. wrap up salmonella magnetic micro-capsule；4. intervene pipeline；
5. liver；6. mechanical arm；7. sophisticated electromagnetism tweezer；8. electromagnetism tweezer iron core is sophisticated；9. solenoid；10 nutrient solutions containing salmonella
Fluid channel is injected with the mixed liquor of Sodium Alginate Hydrogel Films degradation solution；11. phosphoglyceride injects fluid channel；12. sodium alginate
Solution injects fluid channel；13. mineral oil injects fluid channel, 14. liver medium vesselses, 15. tumours, 16. knub positions.
Embodiment
Term " micro-fluidic " in this refers to a kind of accurate control and manipulation minute yardstick fluid, especially refers in particular to sub-micron
The technology of structure.
According to the magnetic of present invention guiding targeting delivering method as shown in figure 1, the delivering method step is as follows；
Step 1: processing the magnetic micro-capsule for including magnetotactic salmonella based on microflow control technique.
Step 2: the method by being intervened in clinical medicine, by the magnetic of the present invention containing magnetotactic salmonella
Micro-capsule is accurately delivered to liver large blood vessel location.
Step 3: the magnetic micro-capsule of the present invention comprising magnetotactic salmonella is being guided into equipment by the magnetic force
As blood flow enters target organ under introduction by magnetic field；The mechanical arm in equipment is guided to adjust the tip location of electromagnetism tweezer by magnetic force,
Along the direction for the blood vessel for leading to tumour, magnetic gradient is formed.Magnetic micro-capsule can be reached along magnetic gradient leads to tumour
Thin vessels leading portion (blood vessel diameter is less than the diameter of magnetic micro-capsule).The locus of magnetic micro-capsule in vivo passes through nuclear-magnetism equipment
(MRI) monitored in real time.
Step 4: in thin vessels leading portion, the magnetic micro-capsule of the present invention containing salmonella passes through inside its own
The degradation solution of parcel ruptures its timing, so that the recombinant attenuated salmonella be discharged；Afterwards, it is just husky into the magnetotactic
Door Salmonella due to the self-driven stage with magnetotaxis, in this stage, the power of the magnetotactic salmonella motion is main
It is that, by self-contained flagellum offer, its direction moved is controlled by magnetic gradient, magnetic gradient covering knub position, so that
Guiding salmonella is moved to knub position, so that the targeting for completing the magnetic carrier containing salmonella is delivered
The distributing procedure of a preferred embodiment of the present invention is as follows：
(1) bacterium solution of attenuation salmonella is inoculated in lysogeny broth (LB) nutrient solution without antibiotic, shaken
Culture is swung to mid log phase, and thalline is collected by centrifugation, cell is washed with the aseptic deionized water of precooling 2 times, it is thin after washing
Bacterium is suspended from the aseptic deionized water of ice precooling, will with electroporation (the Eppendorf electroporations of Multiporator 4308)
PTps-ABG6, pTps-XYZ are transfected to attenuation salmonella, so as to realize the magnetotaxis of attenuation salmonella.
(2) using the soft lithography of semiconductor technology, polydimethylsiloxane (PDMS) fluid channel is processed.Such as
Shown in Fig. 2, the fluid channel includes the mixed liquor injection of the nutrient solution containing salmonella and Sodium Alginate Hydrogel Films degradation solution
Fluid channel 10, phosphoglyceride injection fluid channel 11, sodium alginate soln injection fluid channel 12, and mineral oil injection fluid channel
13.In a preferred embodiment, the depth of the fluid channel is about 100 microns, and exit width is about 400 microns.It will contain
LB nutrient solution (the bacterial densities of attenuation salmonella：1×105Individual/milliliter) and 0.0162 grams per milliliter Sodium citrate
Tribasic dihydrate Sodium Alginate Hydrogel Films degradation solution is mixed, and afterwards, according to shown in Fig. 2, by syringe pump, this is mixed
Close liquid, phosphoglyceride, the marine alga acid solution (magnetic particles comprising ferroferric oxide magnetic nano-particles：0.005 gram/
Milliliter, marine alga acid solutions：0.01 grams per milliliter) and mineral oil be injected separately into the chip, form marine alga acid solution parcel phosphoric acid
Glyceride droplet includes the structure of mixed liquor droplet again.Again, marine alga acid solution forms marine alga acid solution by mineral oil liquid pinch off
Droplet, the microlayer model is injected into calcium chloride solution, and alginic acid droplet will crosslink reaction with calcium chloride solution, forms water-setting
Glue shell, so as to form magnetic microcapsule structure.
(3) method by being intervened in clinical treatment, is accurately delivered to target organs large blood vessel location by magnetic micro-capsule, connects
, magnetic micro-capsule can be under the introduction by magnetic field of the sophisticated electromagnetism tweezer controlled by mechanical arm as blood flow enters target organ.Sophisticated electricity
Magnetic tweezer is made up of a diameter of 1 millimeter of copper cash and cylindrical soft-iron iron core two parts, and copper cash forms 5000 circles around iron core one end
Solenoid, the iron core other end is processed to tip, for converging magnetic field.As shown in figure 1, sophisticated electromagnetism tweezer iron core is basic including one
In the part of cylinder, and it is connected to the basic part in cone of one end of the cylinder.Wherein this is substantially cylindrical
Part about 22 centimetres of length, diameter of section is about 16 centimetres.This be in substantially cone part bed-plate dimension with
The face size of the cylindrical part is essentially identical, can highly design according to actual needs.In drawing for MRI system
Lead down, determine tumour particular location, the tip location of electromagnetism tweezer is adjusted by mechanical arm, along the side for the blood vessel for leading to tumour
To formation magnetic gradient.Magnetic micro-capsule can reach the thin vessels leading portion for leading to tumour along magnetic gradient.Before thin vessels
Section, magnetic micro-capsule ruptures its timing by the degradation solution of its own internal package, so that the attenuation Salmonella in micro-capsule is thin
Bacterium discharges.Afterwards, just into the self-driven stage of magnetotactic bacteria, in this stage, the power of magnetotactic attenuation salmonella motion
Mainly provided by self-contained flagellum, its direction moved is controlled by external magnetic field gradient, and external magnetic field gradient is passed through
Knub position, guiding salmonella is moved to knub position, so as to complete the targeting Delivery of recombinant attenuated salmonella.
Claims (10)
1. a kind of magnetic micro-capsule, including：
Contain in the mixing drop being mixed to get by the nutrient solution and degradation solution of the salmonella containing magnetotactic, the magnetotactic salmonella
Magnetic corpusculum；
Phosphoglyceride droplet, wherein including the mixing drop；And
Magnetic hydrogel shell, it is wrapped in outside the phosphoglyceride droplet.
2. magnetic micro-capsule according to claim 1, it is characterised in that the degradation solution is degraded for Sodium Alginate Hydrogel Films
Liquid.
3. magnetic micro-capsule according to claim 2, it is characterised in that the salmonella containing magnetotactic in the mixing drop
The bacterial density of nutrient solution is 1 × 105Individual/milliliter；The concentration of the Sodium Alginate Hydrogel Films is 0.01 grams per milliliter.
4. magnetic micro-capsule according to claim 1, it is characterised in that the magnetic hydrogel shell is by including four oxidations
The marine alga acid solution and calcium chloride solution of three ferromagnetism nanometer particles crosslink reaction and formed.
5. magnetic micro-capsule according to claim 1, it is characterised in that the magnetic corpusculum is answered by plasmid pTps-ABG6
Fit and plasmid pTps-XYZ complexs are built by electroporation method.
6. the manufacture method of the magnetic micro-capsule according to any one of claim 1-5, comprises the following steps：
Bacterium solution containing salmonella is inoculated in lysogeny broth (LB) nutrient solution without antibiotic, shaken cultivation is to right
Number growth medium, is collected by centrifugation thalline, is washed with the aseptic deionized water of precooling 2 times, the bacterium after washing is suspended from ice precooling
Aseptic deionized water, with electroporation by pTps-ABG6, pTps-XYZ plasmid transfection to salmonella, so as to realize Salmonella
The magnetotaxis of bacterium；
Using the soft lithography of semiconductor technology, microchannel chip is processed；
The Sodium Alginate Hydrogel Films degradation solution of LB nutrient solutions containing the magnetotactic salmonella and 0.0162 grams per milliliter is mixed；
Phosphoglyceride, comprising concentration it is 0.005g/mL ferroferric oxide magnetic nanos grain by the mixed liquor by syringe pump
The marine alga acid solution and mineral oil of son are injected separately into the microchannel chip, form marine alga acid solution parcel phosphoglyceride droplet
The structure of mixed liquor droplet is included again；
Marine alga acid solution forms marine alga acid solution droplet by mineral oil liquid pinch off, and the microlayer model is injected into calcium chloride solution, sea
Alginic acid droplet will crosslink reaction with calcium chloride solution, hydrogel shell be formed, so as to form magnetic microcapsule structure.
7. method according to claim 5, it is characterised in that the depth of the fluid channel is 100 microns.
8. a kind of magnetic guiding delivering method of salmonella, comprises the following steps：
Step 1: manufacture includes the magnetic micro-capsule according to one of claim 1-5 of the salmonella；
Step 2: the magnetic micro-capsule to be delivered to the first position near target location；
Step 3: guiding magnetic micro-capsule described in the introduction by magnetic field of equipment to reach the second place near target location using magnetic force；
Step 4: the magnetic micro-capsule ruptures in the second place, so as to discharge the salmonella；Afterwards, become described in making
Magnetic salmonella is moved to target location under the guiding of magnetic field gradient.
9. method according to claim 8, it is characterised in that the magnetic force guiding equipment includes mechanical arm and electromagnetism tweezer；
The tip location of the electromagnetism tweezer can be adjusted by the mechanical arm, so as to form the magnetic field gradient.
10. method according to claim 8, it is characterised in that in the step 4, the magnetic containing the salmonella
Property micro-capsule by the degradation solution of its own internal package make its timing rupture, so that the salmonella be discharged.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1679518A (en) * | 2005-01-12 | 2005-10-12 | 华东理工大学 | Magnetic medicinal capsules and preparation thereof |
| CN104256631A (en) * | 2014-07-10 | 2015-01-07 | 绵阳罗斯贸易有限公司 | Yacon flour capable of coordinating intestines and stomach, and preparation method thereof |
| CN104666276A (en) * | 2013-11-29 | 2015-06-03 | 康普药业股份有限公司 | Preparation method of 5-Fu magnetic microcapsule |
| CN205434422U (en) * | 2015-12-29 | 2016-08-10 | 何凯 | Drug carrier |
-
2017
- 2017-05-02 CN CN201710301114.3A patent/CN107260703A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1679518A (en) * | 2005-01-12 | 2005-10-12 | 华东理工大学 | Magnetic medicinal capsules and preparation thereof |
| CN104666276A (en) * | 2013-11-29 | 2015-06-03 | 康普药业股份有限公司 | Preparation method of 5-Fu magnetic microcapsule |
| CN104256631A (en) * | 2014-07-10 | 2015-01-07 | 绵阳罗斯贸易有限公司 | Yacon flour capable of coordinating intestines and stomach, and preparation method thereof |
| CN205434422U (en) * | 2015-12-29 | 2016-08-10 | 何凯 | Drug carrier |
Non-Patent Citations (3)
| Title |
|---|
| ISABEL KOLINKO ET AL.: "Biosynthesis of magnetic nanostructures in a foreign organism by transfer of bacterial magnetosome gene clusters", 《NATURE NANOTECHNOLOGY LETTERS》 * |
| TAO SUN ET AL.: "micromanipulation for coiling microfluidic Spun Alginate microfibers by Magnetically Guided System", 《IEEE ROBOTICS AND AUTOMATION LETTERS》 * |
| 廖成水等: "减毒沙门氏菌作为抗肿瘤及肿瘤基因治疗载体的研究进展", 《生物学通报》 * |
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Application publication date: 20171020 |