阅读说明：本技术 一种活体内脊柱包虫病动物模型的构建方法 (Construction method of in vivo echinococcosis animal model ) 是由 王思博 王珊 王维山 戴毅 柯巍 张昊 杨智 于 2019-12-05 设计创作，主要内容包括：本发明涉及脊柱包虫病治疗的技术领域,公开了一种活体内脊柱包虫病动物模型的构建方法,包括如下步骤：步骤一、细粒棘球蚴悬液制备：配置细粒棘球蚴悬液,抽存于注射器中备用；步骤二、术前处理:所有沙鼠麻醉后固定于操作台上,背部操作区域清除毛发；步骤三、体内种植：手指确定椎体序列,以骶髂关节为水平线,沿着棘突向上确定T6-L5阶段,沿着棘突向两侧定位椎旁肌组织,从L4-L5节段斜30°进针,针头进入椎旁肌肉组织后水平进针,至T6-T7节段；步骤四、术后处理：注射后沙鼠饲养一段时间,处死模型后切开背侧皮肤,暴露椎旁肌肉组织,可见大量包虫囊泡聚集。本发明模型成功构建脊柱包虫活体内动物模型,为脊柱包虫的基础研究提供了研究平台。(The invention relates to the technical field of treatment of echinococcosis spine, and discloses a construction method of an in vivo echinococcosis spine animal model, which comprises the following steps: step one, preparation of echinococcus granulosus suspension: preparing a suspension of echinococcus granulosus, and pumping the suspension into an injector for later use; step two, preoperative treatment, namely fixing all gerbils on an operation table after anesthesia, and removing hairs in a back operation area; step three, planting in vivo: determining vertebral body sequence by fingers, determining a T6-L5 stage upwards along a spinous process by taking a sacroiliac joint as a horizontal line, positioning paraspinal muscle tissues along the spinous process to two sides, performing needle insertion at an angle of 30 degrees from an L4-L5 section, and performing horizontal needle insertion after a needle head enters the paraspinal muscle tissues to a T6-T7 section; step four, postoperative treatment: after injection, gerbils were kept for a period of time, and after sacrifice of the model, dorsal skin was incised to expose paraspinal musculature, with a large amount of hydatid vesicle accumulation visible. The model successfully constructs the in-vivo animal model of the hydatid spinalis and provides a research platform for basic research of the hydatid spinalis.)

1. A construction method of an in vivo echinococcosis spine animal model is characterized by comprising the following steps:

step one, preparation of echinococcus granulosus suspension: preparing a suspension of echinococcus granulosus, and pumping the suspension into an injector for later use;

step two, preoperative treatment: all gerbils are fixed on an operation table after anesthesia, hairs are removed from a back operation area, the range of the back area comprises a joint part from a thoracic vertebra (T1) starting position to a far end lumbosacral part, and two sides extend to two sides along a posterior median line by 1.5 cm;

step three, planting in vivo: sterilizing an injection area, paving a towel, wearing sterile gloves, determining a vertebral body sequence by fingers, determining a T6-L5 stage upwards along a spinous process by taking a sacroiliac joint as a horizontal line, positioning paraspinal muscle tissues to two sides along the spinous process, performing needle insertion at an inclination angle of 30 degrees from an L4-L5 section, performing horizontal needle insertion after a needle head enters the paraspinal muscle tissues until the T6-T7 section, sucking blood, slowly pushing fine particle echinococcus suspension, withdrawing the needle along the paraspinal muscles at the same time until a syringe is pulled out, and performing periploid implantation on the paraspinal tissues by the same method;

step four, postoperative treatment: the gerbil is transferred into a relatively clean observation cage for keeping warm, the gerbil is transferred into a feeding cage after the gerbil is completely clear, the gerbil is fed for a period of time after injection, when the back of the gerbil is obviously raised and can touch subcutaneous tumor or the gerbil has abnormal movement and other symptoms, the back skin is cut after the model is killed, paraspinal muscle tissues are exposed, a large amount of hydatid vesicles are visible to be gathered, and the gerbil model shows bone infiltration and damage.

2. The method for constructing an in vivo animal model of echinococcosis spinalis according to claim 1, wherein in the first step, the density of the echinococcus granulosus suspension is 5000 heads/ml.

3. The method for constructing an in vivo animal model of echinococcosis spine according to claim 1 or 2, wherein 10ml is stored in the syringe in the first step.

4. The method for constructing an in vivo echinococcosis spine animal model according to claim 1, wherein in the second step, the gerbil is fixed on the operation table in the prone position.

5. The method for constructing an in vivo animal model of echinococcosis spine according to claim 1, wherein the period of time is 5-6 months in step four.

Technical Field

The invention relates to the technical field of treatment of echinococcosis spine, in particular to a construction method of an in-vivo echinococcosis spine animal model.

Background

Echinococcosis is a common zoonosis disease, which not only harms public health but also affects economic development, and echinococcosis causes damage to single or multiple organs, mainly including liver (70%), lung (20%), and also 10% of all the organs that may be parasitic in any part of the body (such as brain and muscle tissue). Echinococcosis is geographically widespread and mainly concentrated in parts of the continental europe, including the mediterranean region, russia, central asia, the midwest region of china, australia, the united states (particularly the south), and the northeast of africa.

The incidence rate of the echinococcosis is low, the echinococcosis accounts for 0.5-4% of the echinococcosis, the biological behavior of the echinococcosis is more complex compared with the echinococcosis in the liver and the abdominal cavity, and the clinical symptoms are not typical. The spine is a common parasitism part of the hydatid spinalis, and the down-pressing of the thorns and the thorns starts to be located in the spinal cavity, grows slowly, and then spreads along cancellous bone and bone holes, so that the bone is damaged, and pathological fracture is caused. Cysts break through the cortical bone and invade the surrounding soft tissue, resulting in large masses. If the skin is further ulcerated, a long-term unhealed fistula is formed, pus and chafer debris flow out, and chronic pyogenic osteomyelitis can be followed. The initial stage of the disease has no obvious symptoms, pain and numbness can appear along with the development of the disease, the cyst can also press nerves, and the symptoms and physical signs of nerve compression are generated, even the paraplegia is caused.

In the aspect of treatment means, surgery is still the first choice method for treating echinococcosis of the spine, the clinical treatment effect of echinococcosis is greatly improved along with the improvement of the surgical method and the improvement of surgical skills, but the cyst fluid and the hydatid extravasation in the surgery can cause anaphylactic shock and even death, secondary infection of echinococcosis and various serious complications such as residual cavity infection and hydrops are caused due to the leakage and residue of head segments after the surgery, and the postoperative recurrence rate can reach 25 percent at most. The benzimidazole (benzamidine) is the main oral anti-echinococcosis drug designated by WHO, and represents that the drugs are Albendazole (ABZ) and Mebendazole (MBZ), according to clinical reports, the cure rate of the ABZ and MBZ for cystic echinococcosis is only 30%, and the reasons may relate to limited absorption of albendazole in the body, low local blood concentration and unstable curative effect.

Therefore, a high-efficiency treatment method for the echinococcosis is clinically lacking. However, since the basic research field lacks relevant animal models based on the hydatid spinalis, in vivo research on the hydatid spinalis cannot be carried out. The development process and mechanism research of the hydatid in vivo still remain blank, and the research of drug sensitivity experiments and treatment methods in animal models cannot be carried out. Therefore, the construction of a novel live animal model of the echinococcus spinatus is of great importance for the exploration of the echinococcosis spinatus and the research on a treatment scheme.

Disclosure of Invention

The invention provides a construction method of an in vivo echinococcosis animal model, which solves the problems in the background technology.

In order to achieve the purpose, the invention provides the following technical scheme:

a construction method of an in vivo echinococcosis spine animal model comprises the following steps:

step one, preparation of echinococcus granulosus suspension: preparing a suspension of echinococcus granulosus, and pumping the suspension into an injector for later use;

step two, preoperative treatment: all gerbils are fixed on an operation table after anesthesia, hairs are removed from a back operation area, the range of the back area comprises a joint part from a thoracic vertebra (T1) starting position to a far end lumbosacral part, and two sides extend to two sides along a posterior median line by 1.5 cm;

step three, planting in vivo: sterilizing an injection area, paving a towel, wearing sterile gloves, determining a vertebral body sequence by fingers, determining a T6-L5 stage upwards along a spinous process by taking a sacroiliac joint as a horizontal line, positioning paraspinal muscle tissues to two sides along the spinous process, performing needle insertion at an inclination angle of 30 degrees from an L4-L5 section, performing horizontal needle insertion after a needle head enters the paraspinal muscle tissues until the T6-T7 section, sucking blood, slowly pushing fine particle echinococcus suspension, withdrawing the needle along the paraspinal muscles at the same time until a syringe is pulled out, and performing periploid implantation on the paraspinal tissues by the same method;

step four, postoperative treatment: the gerbil is transferred into a relatively clean observation cage for keeping warm, the gerbil is transferred into a feeding cage after the gerbil is completely clear, the gerbil is fed for a period of time after injection, when the back of the gerbil is obviously raised and can touch subcutaneous tumor or the gerbil has abnormal movement and other symptoms, the back skin is cut after the model is killed, paraspinal muscle tissues are exposed, a large amount of hydatid vesicles are visible to be gathered, and the gerbil model shows bone infiltration and damage.

As a preferred technical scheme of the invention, in the first step, the density of the echinococcus granulosus suspension is 5000 heads/ml.

In a preferred embodiment of the present invention, in step one, 10ml is stored in the syringe.

In the second step, the gerbil is fixed on the operating table in the prone position.

As a preferable technical scheme of the invention, in the step four, the period of time is 5-6 months.

The invention has the following advantages:

the model successfully constructs the in-vivo animal model of the hydatid spinalis and provides a research platform for basic research of the hydatid spinalis.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.

FIG. 1 is a flow chart of a construction method of an in vivo echinococcosis spine animal model.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

A construction method of an in vivo echinococcosis spine animal model comprises the following steps:

step one, preparation of echinococcus granulosus suspension: preparing a echinococcus granulosus suspension with a density of 5000 heads/ml, and pumping the echinococcus granulosus suspension into a 10ml syringe for later use;

step two, preoperative treatment: fixing the anesthetized gerbils in prone positions on an operation table, removing hairs in a back operation area, wherein the range of the back area comprises a joint from a thoracic vertebra (T1) starting position to a far-end lumbosacral part, and two sides extend 1.5cm along a posterior median line to two sides;

step three, planting in vivo: sterilizing an injection area, paving a towel, wearing sterile gloves, determining a vertebral body sequence by fingers, determining a T6-L5 stage upwards along a spinous process by taking a sacroiliac joint as a horizontal line, positioning paraspinal muscle tissues to two sides along the spinous process, performing needle insertion at an inclination angle of 30 degrees from an L4-L5 section, performing horizontal needle insertion after a needle head enters the paraspinal muscle tissues to a T6-T7 section, sucking the paraspinal muscle tissues without blood, slowly pushing fine particle echinococcus suspension, withdrawing the needle along the paraspinal muscle at the same time until a syringe is pulled out, and performing fasciola hepatica planting on the paraspinal muscle tissues of the gerbils by the same method;

step four, postoperative treatment: the gerbil is transferred into a relatively clean observation cage, the gerbil is kept warm, the gerbil is transferred into a feeding cage after the gerbil is completely clear, the gerbil is fed for 5-6 months after injection, the dorsal skin of the model is cut after the model is killed, paraspinal muscle tissues are exposed, a large number of hydatid vesicle aggregates can be seen, and the gerbil model shows bone infiltration and destruction.

The invention has the following advantages:

the model successfully constructs the in-vivo animal model of the hydatid spinalis and provides a research platform for basic research of the hydatid spinalis.

It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.