CN102038560A - Construction method of operation model for collecting cerebrospinal fluid repeatedly - Google Patents
Construction method of operation model for collecting cerebrospinal fluid repeatedly Download PDFInfo
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- CN102038560A CN102038560A CN 201010610122 CN201010610122A CN102038560A CN 102038560 A CN102038560 A CN 102038560A CN 201010610122 CN201010610122 CN 201010610122 CN 201010610122 A CN201010610122 A CN 201010610122A CN 102038560 A CN102038560 A CN 102038560A
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Abstract
The invention discloses a construction method of an operation model for collecting cerebrospinal fluid repeatedly, comprising the following steps: positioning an operation drilling hole on the planar structure of an animal skull; puncturing into the drilling hole by an external sleeve puncturing needle; burying and fixing a flow guiding pipe; and collecting the cerebrospinal fluid for repeatedly by the flow guiding pipe. The method provides a new model for collecting the cerebrospinal fluid in a more effective and reasonable manner for a pharmacological experiment of toxic generation and pharmacological generation and the like. According to the method, the survival rate of operation of the animal can reach 80%, the success rate of the model can reach 60%, and thus the construction method of the model is reasonable in design, and is beneficial to collecting the cerebrospinal fluid repeatedly.
Description
Technical field
The present invention relates to cerebrospinal fluid acquisition method field, be specifically related to the construction method that a kind of cerebrospinal fluid is repeatedly gathered surgery models.
Background technology
Cerebrospinal fluid (Cerebrospinal fluid; CSF) be the water white liquid that is filled in each ventricles of the brain, subarachnoid space and the myelocoele; the similar performance of character and blood plasma and lymph fluid; viscosity slightly; be used to supply the certain nutrition of brain cell, transport cerebral tissue metabolite, regulate central nervous system's acid-base balance and cushion brain and the pressure of spinal cord, brain and spinal cord are had protection and support effect.Whether therefore study medicine can effectively can explore by the content of detection of drugs in cerebrospinal fluid by blood brain barrier.
The collection of cerebrospinal fluid is the common operation difficult problem of clinical trial and non-clinical study, brain is mammal vital center and the accumulative place of many important maincenters, so even intracranial is being rich in cerebrospinal fluid, for ensuring the safety of human life, the acquisition method of cerebrospinal fluid is often selected the spinal measurement that punctures in clinical trial.The collection of cerebrospinal fluid at present mostly is disposable collection, and animal model is based on large and small Mus.In non-clinical study, most of experimental selection are thrust from the laboratory animal cervical region with puncture needle, arrive the oblongata pond and gather.
When doing medicine in the research of the intravital dynamic metabolism of machine, the method that the Different Individual animal is gathered in many employings in batches satisfies the drug data of many time points.This replacement method has limited the science of pharmacokinetics/pharmacodynamics (PK/PD) test.Therefore utilizing dog or other animals to set up the non-rodent cerebrospinal fluid of new class repeatedly gathers surgery models research seems particularly important to biological medicine.
Summary of the invention
The invention provides a kind of cerebrospinal fluid and repeatedly gather the construction method of surgery models, this method is simple to operate, is easy to control, has realized the repeatedly collection to cerebrospinal fluid.
A kind of cerebrospinal fluid is repeatedly gathered the construction method of surgery models, may further comprise the steps:
After Animal Anesthesia, positioning operation boring on animal skull planar structure, in described boring through the trocar sheath lancet puncture, imbed drainage tube and fixing drainage tube, by drainage tube collect cerebrospinal fluid repeatedly.
The diameter of described boring should not be too big, to avoid causing excessive wound surface, is preferably 2mm～5mm.
For the assurance puncture needle passes through skull smoothly, and accurately arrive cerebellar fossa, the physiology slit is aimed in described boring, and the direction of boring should be consistent with the puncture direction.
Described boring is preferably placed at the skull planar central.
In pin, fill before the described trocar sheath lancet puncture blunt nosed in pin, syringe needle is put to the hole of having bored, puncture needle thrusts the arrival cerebellar fossa downwards for 30 °～45 ° along the physiology slit during puncture, gos deep into about 3～5cm.
Described puncture needle syringe needle is extracted interior pin after arriving cerebellar fossa, drainage tube is inserted in the pin hole, fixedly drainage tube is slowly extracted puncture needle, with the fixing drainage tube skull outer end portion of medical bio glue, expose several centimetres of drainage tubes in order to collect cerebrospinal fluid, suture muscles and scalp guarantee the drainage tube fixation, and be inner unobstructed.The dog post-operative recovery is more than 12 hours.
Described drainage tube can be selected drain vinyon (PE) pipe for use.
Described animal is non-rodent, is preferably dog.
The time of described collect cerebrospinal fluid is that animal recovered more than 12 hours, and the used device of collect cerebrospinal fluid can be extraction-type devices such as syringe.
Compared with prior art, the present invention has following advantage:
The inventive method has realized the collect cerebrospinal fluid at the cerebellar fossa place, because cerebellar fossa is bigger cavity of intracranial, it is higher relatively that it stores the cerebrospinal fluid ability; And cerebellar fossa is a skull low-lying cavity bottom, and operation boring is above skull, and therefore animal waking state or narcotism cerebrospinal fluid can not run off because of the operation breach.
The inventive method has realized the repeatedly collection to cerebrospinal fluid for pharmacological testings such as poison generation, medicine generation provide more effective and reasonable cerebrospinal fluid collection new model.The operation survival rate of animal can reach 80% in this method, and the model success rate can reach 60%, illustrates that this model building method is reasonable in design, helps the repeatedly collection of cerebrospinal fluid.
Description of drawings
Fig. 1 is the plan structure sketch map of dog head skeleton;
Fig. 2 is the side structure sketch map of dog head skeleton;
Wherein, 1 is frontal bone; 2 is processus temporalis ossis zygomatici; 3 is eye socket; 4 is interparietal bone; 5 is parietal bone; 6 is drill hole; 7 is processus zygomaticus ossis temporalis; 8 is the masseter nest; 9 is the part cerebellar fossa; 10 are the residue cerebellar fossa.
The specific embodiment
After now specific embodiments of the invention further being described in.
As depicted in figs. 1 and 2, the structure of dog head skeleton comprises: frontal bone 1; Processus temporalis ossis zygomatici 2; Eye socket 3; Interparietal bone 4; Parietal bone 5; Drill hole 6; Processus zygomaticus ossis temporalis 7; Masseter nest 8; Cerebellar fossa 9 and 10.
After pentobarbital sodium solution (1ml/kg) anesthesia of Beagle dog intravenous injection 3%, prostrate being fixed on the operating-table.Rise with soft bolster and make occipital bone up at the throat place, fixedly Beagle dog head.With Beagle dog occipital bone is the center, and dog occipital bone left rear side hair is shaved off.Scratch a cross sections about 2cm with scalpel shaving the territory, hair-fields from occipital bone 2～3cm place.Separate top, dog skull plane fascia with operation separation instrument passivity and expose skull top muscle group.Downward separating muscle exposes until skull along the vertical skull of muscle lines plane.Determining that the skull planar central is a drill hole 6, get out the hole of 2～5mm at the center with cranial drill, is the skull plane about diameter 2cm around the hole.For the assurance puncture needle passes through skull smoothly, and accurately arrive cerebellar fossa 9 or 10, boring direction should be consistent with the puncture direction, and the physiology slit is aimed in boring, and cerebellum is the physiology slit with the brain place of link in the dog cerebral tissue.
Use the trocar sheath external diameter to be the 1.6mm puncture needle, in pin, fill before the puncture blunt nosed in pin, syringe needle is put to the hole of having bored, during puncture puncture needle along the physiology slit downwards 30～45 ° thrust the arrival cerebellar fossa, go deep into about 3～5cm.
After piercing needle arrives cerebellar fossa, extract interior pin, can see that cerebrospinal fluid overflows (intracranial pressure effect) from pin hole.Drainage tube is inserted in the pin hole, and fixedly drainage tube is slowly extracted puncture needle, with the fixing drainage tube skull outer end portion of Fibrin Glue, exposes several centimetres of drainage tubes in order to collect cerebrospinal fluid.Operative site is spilled into antibiotic, suture muscles and scalp, and narcotism head portion low level is placed.Seal the drainage tube opening, prevent that cerebrospinal fluid is excessive.The wound care next day of postoperative.
Postoperative 12 hours is opened drainage tube and is sealed, and syringe is inserted drainage tube, extracts 50 μ l liquid in pipe and casts out.Treat that cerebrospinal fluid oozes out from drainage tube voluntarily; Immediately take cerebrospinal fluid slowly to extract with syringe.Postoperative extracts cerebrospinal fluid and be used to check the cerebrospinal fluid success rate of operation every day, observes altogether 7 days.The extraction amount is set to: the 1st day (collection of postoperative narcotism branch) 4.5ml; Clear-headed back 0.5ml; Extract 0.5ml all the other 13 day every days.
Success rate of operation is as the criterion with the survival of Beagle dog postoperative health.But the model success rate is not successful standard by the blood contamination cerebrospinal fluid by postoperative Beagle dog continuous acquisition.10 Beagle dog operations, success rate of operation 80%.Operation back collect cerebrospinal fluid every day all can collect cerebrospinal fluid in 7 days smoothly, and the model success rate is 60%.
Embodiment 2
Method makes up 4 of animal patterns as described in example 1 above.Get medicine ethylenediamine (EDA) injection of certain penetrable blood brain barrier, it is that the speed intravenous drip of pressing medicine 1mg/kg/h is to 3h in 0.9% the NaCl injection that medicine is disposed at mass percentage concentration.And after administration 0,30,60 and 180min (administration end) and drug withdrawal 0.5,1,2,3 and 4h regularly take 2ml whole blood and 0.3ml cerebrospinal fluid.Use the EDA concentration in the method detection of biological sample of being set up.Use DAS 2.0 pharmacokinetics software analysis results.By the pharmacokinetics result as can be known, give Beagle dog 3mg/kg medicine in the intravenous drip mode after, EDA eliminates in the mode of three-compartment model.
Claims (9)
1. a cerebrospinal fluid is repeatedly gathered the construction method of surgery models, may further comprise the steps:
After Animal Anesthesia, positioning operation boring on animal skull planar structure, in described boring through the trocar sheath lancet puncture, imbed drainage tube and fixing drainage tube, by drainage tube collect cerebrospinal fluid repeatedly.
2. cerebrospinal fluid according to claim 1 is repeatedly gathered the construction method of surgery models, it is characterized in that, the diameter of described boring is 2mm～5mm.
3. cerebrospinal fluid according to claim 1 is repeatedly gathered the construction method of surgery models, it is characterized in that, the physiology slit is aimed in described boring, and the direction of boring is consistent with the puncture direction.
4. cerebrospinal fluid according to claim 1 is repeatedly gathered the construction method of surgery models, it is characterized in that, described boring is positioned at the skull planar central.
5. cerebrospinal fluid according to claim 1 is repeatedly gathered the construction method of surgery models, it is characterized in that, in pin, fill before the described trocar sheath lancet puncture blunt nosed in pin, syringe needle is put to the hole of having bored, puncture needle thrusts the arrival cerebellar fossa downwards for 30 °～45 ° along the physiology slit during puncture, gos deep into 3cm～5cm.
6. cerebrospinal fluid according to claim 1 is repeatedly gathered the construction method of surgery models, it is characterized in that, described trocar sheath puncture needle syringe needle is extracted interior pin after arriving cerebellar fossa, drainage tube is inserted in the pin hole, and fixedly drainage tube is slowly extracted puncture needle, uses fixedly drainage tube skull outer end portion of medical bio glue, expose several centimetres of drainage tubes in order to collect cerebrospinal fluid, suture muscles and scalp guarantee the drainage tube fixation, and be inner unobstructed.
7. cerebrospinal fluid according to claim 1 is repeatedly gathered the construction method of surgery models, it is characterized in that, described animal is non-rodent.
8. cerebrospinal fluid according to claim 7 is repeatedly gathered the construction method of surgery models, it is characterized in that, described animal is a dog.
9. cerebrospinal fluid according to claim 1 is repeatedly gathered the construction method of surgery models, it is characterized in that, the time of described collect cerebrospinal fluid is that animal recovers more than 12 hours.
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| CN 201010610122 CN102038560A (en) | 2010-12-28 | 2010-12-28 | Construction method of operation model for collecting cerebrospinal fluid repeatedly |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102784012A (en) * | 2012-03-16 | 2012-11-21 | 中山大学 | Blood brain barrier pharmacokinefic continuous dosing system and detecting system |
| CN103598924A (en) * | 2013-12-03 | 2014-02-26 | 菏泽学院 | Experiment rat central nervous sucking and pulling device |
| CN104323868A (en) * | 2014-11-06 | 2015-02-04 | 中国人民解放军第四军医大学 | Micro-infusion method and device thereof based on epididymis duct intracavity environment experiment |
| CN117224267A (en) * | 2023-11-10 | 2023-12-15 | 北京伽拓医药研究有限公司 | Method for establishing canine frontal sinus defect and cerebrospinal fluid rhinorrhea model and application thereof |
-
2010
- 2010-12-28 CN CN 201010610122 patent/CN102038560A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102784012A (en) * | 2012-03-16 | 2012-11-21 | 中山大学 | Blood brain barrier pharmacokinefic continuous dosing system and detecting system |
| CN102784012B (en) * | 2012-03-16 | 2015-05-06 | 中山大学 | Blood brain barrier pharmacokinefic continuous dosing system and detecting system |
| CN103598924A (en) * | 2013-12-03 | 2014-02-26 | 菏泽学院 | Experiment rat central nervous sucking and pulling device |
| CN103598924B (en) * | 2013-12-03 | 2015-09-09 | 菏泽学院 | Experimental rat nervus centralis draws drawing device |
| CN104323868A (en) * | 2014-11-06 | 2015-02-04 | 中国人民解放军第四军医大学 | Micro-infusion method and device thereof based on epididymis duct intracavity environment experiment |
| CN117224267A (en) * | 2023-11-10 | 2023-12-15 | 北京伽拓医药研究有限公司 | Method for establishing canine frontal sinus defect and cerebrospinal fluid rhinorrhea model and application thereof |
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Application publication date: 20110504 |