CN110755642A - Composite coloring agent and application thereof - Google Patents
Composite coloring agent and application thereof Download PDFInfo
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- CN110755642A CN110755642A CN201911278140.4A CN201911278140A CN110755642A CN 110755642 A CN110755642 A CN 110755642A CN 201911278140 A CN201911278140 A CN 201911278140A CN 110755642 A CN110755642 A CN 110755642A
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- toluidine blue
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- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/006—Biological staining of tissues in vivo, e.g. methylene blue or toluidine blue O administered in the buccal area to detect epithelial cancer cells, dyes used for delineating tissues during surgery
Abstract
The invention discloses a composite coloring agent and application thereof, wherein the composite coloring agent is a mixed solution consisting of an acetic acid solution, a toluidine blue solution and normal saline, the mass concentration of acetic acid in the mixed solution is 1.5-2%, and the mass concentration of toluidine blue in the mixed solution is 0.2-0.4%. The complex staining agent of the invention can be matched with a gastrointestinal scope, so that the focus which is difficult to distinguish under white light can be conveniently found, and the complex staining agent is particularly used for treating canine esophagitis and can improve the accurate diagnosis rate and sensitivity of the canine esophagitis.
Description
Technical Field
The invention relates to the technical field of biochemistry, and particularly relates to a composite coloring agent and application thereof.
Background
The rapid development of the pet medical industry has higher and higher requirements on pet disease diagnostic instruments. The diagnosis of esophageal diseases of pet dogs has become a great problem in the pet diagnosis and treatment industry. Many dogs do not show obvious clinical symptoms when suffering from esophageal injury, are difficult to find esophageal diseases by X-ray examination, cannot make accurate disease diagnosis, and have the danger of choking into the trachea and suffocating when being drenched with barium meal or iodine preparation contrast agents.
Canine esophagitis generally refers to inflammation of the esophagus caused by edema and congestion of the esophageal mucosa due to stimulation or damage to superficial or deep tissues of the esophageal mucosa. Chemical stimuli include gastric acid, bile, alcohol, strong acids, strong bases, drugs, etc.; physical stimulation includes food with high temperature, sharp foreign bodies, esophageal foreign bodies (such as fish bones) incarceration, long-term placement of nasogastric feeding tubes and the like; in pet clinic, dog esophagitis is common in improper feeding management of pet owners, and excessively sharp food (such as bones and the like) is fed to pet dogs, so that esophagitis is caused after the esophagus of the dogs is scratched and stimulated by sharp foreign matters. In addition, in the clinical treatment process of certain diseases, esophagitis is caused by local damage of the esophagus due to long-term treatment (doxycycline and the like) of certain medicines. The diagnosis of esophagitis of pet dogs is always a great problem in the pet diagnosis and treatment industry, and the clinical symptoms of esophageal diseases mainly include reflux, dysphagia, vomiting, appetite reduction and the like. The clinical symptoms of canine esophagitis have high similarity with the clinical symptoms of canine digestive tract diseases, and are difficult to distinguish from other canine digestive tract diseases. In addition, the canine esophagitis has unobvious clinical symptoms and no specificity, and is easy to be missed or misdiagnosed in the clinical diagnosis of pets. Therefore, accurate diagnosis of canine esophagitis has been a difficult problem in clinical diagnosis of pets.
The gastrointestinal endoscope is a better diagnostic instrument in clinical diagnosis of pets, is painless and harmless to a sick dog like other imaging instruments (such as digestive tract radiography, ultrasound, CT, MR and the like), but has better directness and timeliness in gastrointestinal endoscope diagnosis. The gastrointestinal endoscope enables a clinical pet doctor to directly observe the instant appearance state of the canine lesion part through screen imaging, and provides great help for disease diagnosis. However, the history of dog performing gastrointestinal endoscopy is not long, and at present, pet gastrointestinal endoscopy is mostly used for taking out foreign matters in pet digestive tracts, and is rarely used as a disease diagnosis method, which is caused by the reason that the digestive tract lesions of pets under white light gastrointestinal endoscopy are not easy to be found, and the like, and is also a great neglect of the function of endoscopic diagnosis and examination.
Disclosure of Invention
Aiming at the prior art, the invention aims to provide a composite coloring agent and application thereof, and the composite coloring agent is matched with a gastrointestinal endoscope, so that a focus which is difficult to distinguish under white light can be conveniently found, and the diagnosis rate and the sensitivity are high particularly for canine esophagitis.
In order to achieve the purpose, the invention adopts the following technical scheme:
the composite coloring agent is a mixed solution composed of an acetic acid solution, a toluidine blue solution and normal saline, wherein the mass concentration of acetic acid in the mixed solution is 1.5% -2%, and the mass concentration of toluidine blue in the mixed solution is 0.2% -0.4%.
Preferably, the mixed solution contains acetic acid at a concentration of 1.5% by mass and toluidine blue at a concentration of 0.2% by mass.
In a second aspect of the present invention, a preparation method of the composite coloring agent is provided, which comprises the following steps: mixing acetic acid solution, toluidine blue solution and normal saline to obtain the composite coloring agent.
Preferably, when the composite coloring agent is prepared, the mass concentration of the adopted acetic acid solution is 3% -4%, and the mass concentration of the toluidine blue solution is 1% -2%.
More preferably, the acetic acid solution and the toluidine blue solution are prepared by using normal saline as a solvent when the composite coloring agent is prepared. The mass concentration of sodium chloride in the physiological saline is 0.9%.
Preferably, the volume ratio of the acetic acid solution, the toluidine blue solution and the physiological saline used for preparing the composite coloring agent is 5:2: 3.
In a third aspect of the invention, the application of the composite staining agent in preparing a canine esophageal mucosa staining reagent is provided.
In a fourth aspect of the invention, the application of the composite staining agent in preparing a dog esophagitis diagnostic reagent is provided.
In a fifth aspect of the invention, there is provided a method of staining canine esophageal mucosa, comprising the steps of: spraying the composite staining agent into dog esophagus, sucking out the residual composite staining agent in the esophagus after 2-3min, and washing the esophagus.
Preferably, the esophagus is washed repeatedly for 3-4 times by using purified water.
Preferably, the volume of each spray of the composite coloring agent is 10-15 ml.
The invention has the beneficial effects that:
1. by combining the dyeing of the composite coloring agent with the gastrointestinal endoscope, the defect that fine focuses under the gastrointestinal endoscope are difficult to observe can be overcome, so that the disease focuses can be visually observed in an enlarged manner, the focuses are highlighted and easier to find after dyeing, and clinical pet doctors are guided to carry out correct diagnosis, so that the endoscope can play a greater role, the sensitivity and specificity of a diseased part in gastrointestinal endoscope examination are increased through dyeing, and the accuracy of disease diagnosis is improved. The dyed esophagus structure level, form and damage degree are easier to observe, and then clinical pet doctors are helped to better judge the real-time condition of the canine esophagus so as to make more accurate diagnosis. The focus which is difficult to distinguish under white light can be conveniently found, biopsy can be more accurately guided, and the detection rate of lesion and the accuracy of diagnosis are improved.
2. The composite staining agent disclosed by the invention is used for staining the canine esophagus and matching with the gastrointestinal endoscopy examination, the diagnosis rate of canine esophagitis is 94.44%, the sensitivity is 92.17%, and compared with the white light gastrointestinal endoscopy examination of the same group, the diagnosis rate is increased by 69.99%, and the sensitivity is increased by 64.59%. The composite coloring agent is a mixed solution prepared by mixing an acetic acid solution, a toluidine blue solution and normal saline in advance, namely the acetic acid solution and the toluidine blue solution are added simultaneously during dyeing, and if the acetic acid or the toluidine blue is added firstly for dyeing or the concentration of the acetic acid or the toluidine blue is changed, the diagnosis rate and the sensitivity are influenced.
Drawings
FIG. 1 is a line graph of staining and white light gastrointestinal mirror scoring in the acetic acid staining group experiment of the present invention;
FIG. 2 is a diagram of esophageal mucosa damage under a white light gastroenterology scope in an acetic acid staining group experiment of the invention;
FIG. 3 is a diagram showing the damage of the mucosa of the esophagus under the acetic acid staining enteroscope in the acetic acid staining group experiment of the present invention;
FIG. 4 is a line graph showing staining and white light gastrointestinal mirror scoring in the toluidine blue staining group experiment of the present invention;
FIG. 5 is a chart of esophagitis mucosa under a white light gastroscope in a toluidine blue staining group experiment of the present invention;
FIG. 6 is a chart of toluidine blue stained gastrointestinal esophagitis mucosa in a toluidine blue stained group experiment of the present invention;
FIG. 7 is a diagram of esophageal mucosa injury under a white light gastroscope in an acetic acid-toluidine blue complex staining group experiment of the present invention;
FIG. 8 is a graph of the damage to the esophageal mucosa under a gastroscope stained with acetic acid-toluidine blue in the acetic acid-toluidine blue complex staining group experiment of the present invention;
FIG. 9 is a diagram of esophageal mucosa injury under a white light gastroscope in a toluidine blue-acetic acid complex staining group experiment of the present invention;
FIG. 10 is a graph of toluidine blue-acetic acid stained gastrointestinal endoscopic esophageal mucosa injury in a toluidine blue-acetic acid complex staining group experiment of the present invention;
FIG. 11 is a graph of esophageal mucosa damage under a white light gastroscope in a mixed acetic toluidine blue staining group experiment according to the present invention;
FIG. 12 is a graph of mixed acetic toluidine blue stained gastrointestinal endoscopic esophageal mucosa injury in a mixed acetic toluidine blue stained group experiment of the present invention;
FIG. 13 is a graph showing the comparison of the diagnosis rate of esophagitis in dogs using the staining methods of comparative examples 1 to 5 and application example 1, respectively, according to the present invention;
FIG. 14 is a comparison graph showing the improvement of the diagnosis rate of canine esophagitis by the staining method of comparative examples 1 to 5 and application example 1, respectively;
FIG. 15 is a graph showing the sensitivity of the present invention to dog esophagitis scores using the staining methods of comparative examples 1 to 5 and application example 1, respectively;
FIG. 16 is a graph showing the improvement of the sensitivity of the staining method of comparative examples 1 to 5 and application example 1 to the canine esophagitis score according to the present invention;
FIG. 17 is a photograph of normal esophageal mucosa under a white light gastroenteroscopy; normal mucous membranes appear as a pale pink white of uniform hue;
FIG. 18 is a picture of normal esophageal mucosa under an acetic acid-stained mirror with a mass concentration of 1.5%; normal mucous membrane is more prominent on the surface, and the phenomenon that the mucous membrane is reddish after whitening exists;
FIG. 19 is a photograph of a white light gastroenteroscopic canine abdominal esophagus; the junction of the esophagus and the cardia of the stomach can be seen, the esophagus at the abdominal part is even and light pink, and the junction of the normal esophagus and the stomach can be seen when the esophagus turns white circularly at the cardia of the stomach;
FIG. 20 is a picture of the junction of the abdominal esophagus and the cardia of the stomach of a canine under an acetic acid staining enteroscopy with a mass concentration of 1.5%; the boundary of the cardia of the stomach and the esophagus of the abdominal part can be clearly seen, the whitening of the esophagus of the abdominal part is reddish after 1.5 percent acetic acid dyeing, and the whitening of the cardia of the stomach is more obvious and the phenomenon of squamazation and stenosis appears;
fig. 21 is a picture of normal esophageal mucosa under a gastroscope stained with toluidine blue at a mass concentration of 0.2%; normal mucous membranes appear as light blue with a uniform hue.
Detailed Description
It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.
As introduced in the background art, because the focus of the digestive tract of the pet under the white-light gastrointestinal endoscope is not easy to be found, the current gastrointestinal endoscope of the pet is mostly used for taking out foreign matters in the digestive tract of the pet and rarely used as a disease diagnosis method; based on the above, the invention provides the composite staining agent, through the matching of the staining of the composite staining agent and a gastrointestinal scope, the focus which is difficult to distinguish under white light can be conveniently found, the biopsy can be more accurately guided, the detection rate of lesion and the accuracy rate of diagnosis are improved, and the diagnosis confirmation rate and the sensitivity are high especially for canine esophagitis.
In order to make the technical solutions of the present application more clearly understood by those skilled in the art, the technical solutions of the present application will be described in detail below with reference to specific embodiments.
The test materials used in the examples of the present invention were all commercially available materials that are conventional in the art and are commercially available.
Example 1: preparation of composite coloring agent
Mixing 3% by mass of acetic acid solution, 1% by mass of toluidine blue solution and 0.9% by mass of sodium chloride normal saline according to the volume ratio of 5:2:3 to prepare the composite coloring agent, wherein the concentration of acetic acid in the prepared composite coloring agent is 1.5%, and the concentration of toluidine blue is 0.2%.
Example 2: preparation of composite coloring agent
Mixing an acetic acid solution with the mass concentration of 4%, a toluidine blue solution with the mass concentration of 2% and a normal saline with the mass concentration of sodium chloride of 0.9% according to the volume ratio of 5:2:3 to prepare a composite coloring agent, wherein the concentration of the acetic acid in the prepared composite coloring agent is 2%, and the concentration of the toluidine blue is 0.4%.
Example 3: preparation of composite coloring agent
Mixing 3.5% acetic acid solution, 1.5% toluidine blue solution and 0.9% sodium chloride normal saline according to the volume ratio of 5:2:3 to prepare the composite coloring agent, wherein the concentration of acetic acid in the prepared composite coloring agent is 1.75%, and the concentration of toluidine blue is 0.3%.
Application example 1: the esophageal mucosa of the experimental dog is stained by the composite stain prepared in the example 1.
Application example 2: the esophageal mucosa of the experimental dog is stained by the composite stain prepared in the example 2.
Application example 3: the esophageal mucosa of the experimental dog is stained by the composite stain prepared in the example 3.
Comparative example 1: and (3) taking an acetic acid solution with the mass concentration of 1.5% as a staining agent to stain the esophageal mucosa of the experimental dog.
Comparative example 2: and (3) dyeing the esophageal mucosa of the experimental dog by using a toluidine blue solution with the mass concentration of 0.2% as a dyeing agent.
Comparative example 3: and adopting Lugol iodine solution with the mass concentration of 1% as a staining agent to stain the esophageal mucosa of the experimental dog.
Comparative example 4: firstly, 1.5% acetic acid solution is adopted for dyeing, and then 0.2% toluidine blue solution is adopted for dyeing the esophageal mucosa of the experimental dog.
Comparative example 5: the method comprises the steps of firstly adopting toluidine blue solution with the mass concentration of 0.2% to dye, and then adopting acetic acid solution with the mass concentration of 1.5% to dye the esophageal mucosa of the experimental dog.
The acetic acid solution and the toluidine blue solution used in each example were prepared using a normal saline solution with a sodium chloride mass concentration of 0.9% as a solvent.
Experiments were conducted to determine the accuracy of the diagnosis results of canine esophagitis after staining canine esophageal mucosa using the methods of application example 1, comparative example 2, comparative example 3, comparative example 4, and comparative example 5, respectively.
Gastrointestinal diagnostic technique using composite staining agent
① anesthesia, because esophagus gastrointestinal microscopic examination observation is needed, experimental examination animals need to be anesthetized, large dogs are fasted for 12-24h before operation and are forbidden for 8h, small dogs are fasted for 8-12h before operation and are forbidden for 6-8h before operation and are subcutaneously injected with atropine injection liquid 20 minutes before anesthesia, 0.04mg/kg im reduces salivary secretion, because atropine can reduce glandular secretion and dry airway, keep respiratory tract smooth and slow gastrointestinal peristalsis, keep visual field clear and convenient operation, prevent heart rate slowing problem caused by medicament and examination operation by blocking vagus nerve reflex, reduce adverse reaction, ensure safety and provide convenient conditions for gastrointestinal microscopic examination, more different anesthesia schemes can be used in clinic, a method of using the xylazine hydrochloride injection liquid (fast new) and the Shutai compound anesthesia is selected in the experiment for anesthesia, the xylazine injection liquid is firstly used for 0.04-0.08ml/kg im and then is diluted by 50 times of the Shutai injection liquid for maintaining anesthesia, and only the adrenalin the intravenous injection operation and the monitoring of dexamethasone is performed in the standby examination.
② posture, the right side of the experimental dog is fixed when the oesophagoscope is used for dyeing, the mouth opener for the dog is arranged between the upper and lower dog teeth on the right side of the experimental dog to ensure that the oral cavity of the dog to be checked is completely opened, the tongue of the dog to be checked is pulled out of the oral cavity, saliva secreted by the dog to be checked is easy to flow out of the oral cavity, and the oral cavity of the dog to be checked is cleaned by gauze in time to maintain the clean oral cavity so as to keep the respiratory tract of the dog to be checked unobstructed.
When the right side position is kept, the neck section esophagus is parallel to the trachea level, and the chest section esophagus is located above the trachea and is parallel to the trachea, so the left side position is kept and the esophagus is not influenced by the gravity compression of the trachea, saliva secreted by the experimental dog in the operation is not easy to flow into the trachea, and the foreign body pneumonia is avoided.
On the left side, similar to the right side, but in the chest region, the esophagus is located below the trachea, which is more difficult to operate than the right side.
In the normal position, the esophagus is relatively lower due to the gravity compression of the trachea, and the esophagus is not easy to inflate to observe the focus after compression.
③ dyeing, wherein the best shooting time is that the esophagus trend is clear, the foam is less, the damaged part is in the center of the visual field, before dyeing, air is firstly sent into the esophagus to display the esophagus trend and the outline, the esophageal focus area is found, a proper shooting angle is found, the esophageal focus picture is shot under a white light gastroenteroscope, after the white light gastroenteroscope shooting is finished, the esophagus is dyed, a proper amount of dye liquid is sprayed into the esophagus, the esophageal focus is observed after a few minutes, the esophagus is aerated again to suck out the residual dye liquid, whether the esophagus is washed or not is selected according to the property of the dye liquid, and the picture after the esophageal focus is dyed is shot.
④ quality evaluation, the position of the esophageal focus is the same when shooting, the esophageal focus is required to be positioned at the center of the picture as much as possible on the picture and kept clear, and the dyeing effect is analyzed by comparing the pictures of the same esophageal focus before and after dyeing, such as the focus size, the dyeing color depth, the range and the like.
⑤ examination procedure for evaluating the severity of esophageal diseases, standardized examination procedures such as lesion size, staining depth, and hematology examination are used.
Method for evaluating esophageal diseases by composite stain enterogastroscope diagnostic technology
Three unknown veterinarians judge whether the dog has the canine esophagitis or not by video in the esophagus of the dog under the same gastrointestinal endoscope. The video that the veterinarian watched requires to be clear, and the video requires that the visible dog esophagus is inflated to be opened, and the video duration is required to be 15-20s when the video starts from the neck esophagus and advances to the abdominal esophagus along the esophageal wall and the video ends when the lens reaches the cardia position. The gastrointestinal observation method comprises the following steps: the observation of the endoscope is combined with the observation of the endoscope, and the original state of the focus is observed mainly by the observation of the endoscope. The perspective and the close-up are combined, the perspective is taken as the main point, the focus panorama is viewed, and the focus structure is viewed closely.
Three veterinarians watch videos and photos under the canine normal esophageal mucosa white-light enteroscope and various staining enteroscopes in advance to know the morphological structure of the canine normal esophageal mucosa. This has some help to their diagnosis of gastroenteroscopic canine esophagitis. Evaluation criteria are specified here for Visual inspection after acetic acid staining (VIA), Visual inspection after toluidine blue staining (VITB) iodine staining (Visual inspection with Lugol's iodine, VILI):
diagnostic criteria for VIA: the canine esophageal mucosa surface becomes uniform pink white and is negative; the canine esophageal mucosa has abnormal bulges which are positive; the canine esophageal mucosa part is not whitened or the whitening unevenness is positive.
Diagnostic criteria for VITB: the canine esophageal mucosal surface appears to be uniform and non-staining or light blue is negative; the canine esophageal mucosa is dyed blue, and the canine esophageal mucosa is positive; the canine esophageal mucosa is stained with a gem blue dot which is positive; the canine esophageal mucosa is dyed dark blue and has blue dots as strong positive;
VILI diagnostic standard: the canine esophageal mucosa appeared uniform brown negative; the canine esophageal mucosa is negative for a part of the area where the canine esophageal mucosa appears, i.e. light brown.
The diagnosis rate (number of confirmed dogs/total number of dogs) × 100%.
Sensitivity was determined blindly by three veterinarians by statistical analysis of the oesophagitis scores obtained from the pictures obtained from the experiments. The sensitivity after staining was relative to that under a white light gastroscope, the procedure was as follows: pictures of an oesophagitis model dog are taken under white light gastroenteroscopy examination and staining enteroscopy examination, the pictures are respectively given to three unknown veterinarians for carrying out oesophagitis distinguishing and scoring, and each picture is given a score of 0-10 points according to the positive score of the oesophagitis from low to high. The veterinarian gives a score by comparing the information given by the picture (e.g. changes in esophageal mucosa morphology, color, etc.) with the normal esophageal mucosa morphology. During the gastrointestinal endoscopy process of each esophagitis model dog, 5 diseased parts are firstly shot under a white light gastrointestinal endoscope, and then 5 diseased parts are shot under the gastrointestinal endoscope after staining. Each veterinarian gave each esophagitis model dog 5 white light and 5 staining underwent gastrointestinal scoring, and finally the scores of each group were collected and the average of each group was taken as the final score. 10 points correspond to a sensitivity of 100%, so sensitivity is the average of canine esophagitis scores × 10 × 100%.
Test animals: 36 healthy laboratory beagle dogs, 18 dogs each for male and female, 12 months of age and similar body types are provided by a veterinary clinical diagnosis laboratory. The animal model is established, and the canine esophagitis is caused by direct esophageal injury to 36 healthy experimental beagle dogs. After the experimental beagle dog is anesthetized, the right side of the experimental beagle dog is horizontally fixed, after the mucosa wall at the same part is repeatedly clamped by a biopsy forceps under a gastrointestinal endoscope to cause mucosa injury, 20ml of hydrogen peroxide is injected into the esophagus immediately to cover the injury part, and the residual liquid is sucked out after the hydrogen peroxide stays in the esophagus for 10 min. Feeding dog food with rough mouthfeel during the establishment period of an esophageal injury model of an experimental dog. After 2 days, if the experimental beagle dog shows symptoms (such as eating slowness, dysphagia, vomiting and the like) and white blood cells rise in blood routine examination, the canine esophagitis model is considered to be successfully established.
An experimental instrument: shanghai adult transit electron enteroscope GVE-2100P; merrill BC-2600 automatic blood cell analyzer; idel dry biochemical analyzer; CARDELL 9500HD ECG monitor; a urine analyzer.
Experimental equipment: 4 mouth openers for dogs, 4 handles of tongue forceps, 5 boxes of indwelling needles, 10 bandages and 2 boxes of self-adhesive tape.
Experimental drugs: atropine, sultam, propofol, simethicone, lidocaine, 3% acetic acid, 1% toluidine blue and the like.
Photographs of normal esophageal mucosa under a white-light gastroenteroscopy and a stained gastroenteroscopy are respectively taken before modeling, and as shown in fig. 17-21, the effect of the normal esophageal mucosa under white light and the effect of staining can be clarified.
The test method comprises the following steps:
1. single staining experiment
18 beagle dogs were divided into 3 groups of 6 dogs each with 3 dogs each for male and female. The first group was subjected to the dyeing test using the method of comparative example 1 and was designated as the acetic acid dyeing group; the second group was subjected to the dyeing test using the method of comparative example 2 and was designated as toluidine blue dye group; thirdly, a dyeing experiment is carried out by adopting the method of a comparative example 3 and is marked as a Lugol iodine solution dyeing group;
the single staining experiment of each group is divided into two parts, wherein the former part is observed under a white light gastrointestinal endoscope, and the latter part is observed under the white light gastrointestinal endoscope after the white light gastrointestinal endoscope observation is finished. The white light gastroscope observation picture score is used as a control group, and the staining gastroscope observation picture score is used as an experimental group.
(1) Acetic acid staining group experiment
Experiment 6 successful beagle dogs built with esophagitis models were subjected to enteroscopy staining experiments, which were numbered AA1, AA2, AA3, AA4, AA5, and AA6, respectively.
Control group: fasting is carried out for 8-12h before general anesthesia, and water is forbidden for 6-8 h. 2 simethicone tablets are taken 1h before anesthesia, and atropine injection (0.04mg/kg) is subcutaneously injected 20min before anesthesia to reduce salivary secretion. After the esophagitis experimental dog is anesthetized, the right side is laid horizontally for fixation, a mouth gag for dogs is arranged between right side dog teeth, a gastrointestinal endoscope is inserted into the esophagus of the dogs to observe the mucous membranes of the neck section, the chest section and the abdomen section of the esophagus, the suspicious lesion area 5 is shot by observation and recording, and the picture of the suspicious lesion area is scored.
Experimental groups: after the white light gastrointestinal endoscopy examination is finished, the same experimental dog inserts a plastic catheter with an opening into an esophagus cavity through a gastrointestinal endoscopy biopsy hole, 10ml of acetic acid with the mass concentration of 1.5% is sprayed from front to back, so that the acetic acid is uniformly distributed on the surface of a mucous membrane, the residual staining solution is sucked out after 1min, the gastrointestinal endoscopy is inserted into the esophagus of the dog to observe the neck section of the esophagus, the mucous membranes of the chest section and the abdomen section, 5 suspicious lesion areas are observed and recorded and shot, and pictures of the suspicious lesion areas are scored.
(2) Toluidine blue staining group experiment
Experiment results were obtained by performing enteroscopy staining experiments on 6 beagle dogs successfully established with an esophagitis model, which were respectively numbered TB1, TB2, TB3, TB4, TB5 and TB 6.
Control group: the method was performed in the same manner as in the control group of acetic acid staining experiment.
Experimental groups: after the white light gastrointestinal endoscope examination is finished, the same experimental dog inserts a plastic catheter with an opening into an esophageal cavity through a gastrointestinal endoscope biopsy hole, 10ml of toluidine blue dye solution with the mass concentration of 0.2% is sprayed from front to back to enable the toluidine blue to be uniformly distributed on the surface of a mucous membrane, the dye solution is sucked out after 3min, and the dyed mucous membrane is repeatedly washed by water for 3-4 times to enable the residual dye solution and liquid in the esophageal cavity to be fully sucked out. After the washing, visit the intestines and stomach mirror and observe the mucous membrane of esophagus neck section, chest section and abdomen section in the dog esophagus, observe and take 5 departments of suspicious lesion area with the record, grade suspicious lesion area picture.
(3) Lugol's iodine staining group experiment
Control group: the method was performed in the same manner as in the control group of acetic acid staining experiment.
Experimental groups: after the white light gastrointestinal endoscopy examination is finished, the same experimental dog is inserted into a plastic catheter with an opening to an esophageal cavity through a gastrointestinal endoscopy biopsy hole, 10ml of Lugol iodine solution dye solution with the mass concentration of 1% is sprayed from front to back, the Lugol iodine solution is uniformly distributed on the surface of a mucous membrane, the dye solution is sucked out after 3min, and the dyed mucous membrane is repeatedly washed by water for 3-4 times, so that the residual dye solution and liquid in the esophageal cavity are fully sucked out. After the washing, visit the intestines and stomach mirror and observe the mucous membrane of esophagus neck section, chest section and abdomen section in the dog esophagus, observe and take 5 departments of suspicious lesion area with the record, grade suspicious lesion area picture.
2. Composite dyeing experiment
18 beagle dogs were divided into 3 groups of 6 dogs each with 3 dogs each for male and female. The first group was subjected to the dyeing test using the method of comparative example 4 and was designated as the acetic acid-toluidine blue complex dyeing group; the second group was subjected to the dyeing test using the method of comparative example 5 and was designated as toluidine blue-acetic acid composite dyeing group; the third group was subjected to the dyeing test in the manner of application example 1 and was designated as a mixed acetic toluidine blue dye group.
The composite staining experiment of each group is divided into two parts, wherein the former part is observed under a white light gastrointestinal endoscope, and the latter part is observed by a staining gastrointestinal endoscope after the white light gastrointestinal endoscope observation is finished. The white light gastroscope observation picture score is used as a control group, and the staining gastroscope observation picture score is used as an experimental group.
(1) Acetic acid-toluidine blue complex staining group experiment
Experiment 6 successful beagle dogs built with esophagitis models are subjected to enteroscopy staining experiments and are respectively numbered as AA-TB1, AA-TB2, AA-TB3, AA-TB4, AA-TB5 and AA-TB 6.
Control group: fasting is carried out for 8-12h before general anesthesia, and water is forbidden for 6-8 h. 2 simethicone tablets are taken 1h before anesthesia, and atropine injection (0.04mg/kg) is subcutaneously injected 20min before anesthesia to reduce salivary secretion. After the esophagitis experimental dog is anesthetized, the right side is laid horizontally for fixation, a mouth gag for dogs is arranged between right side dog teeth, a gastrointestinal endoscope is inserted into the esophagus of the dogs to observe the mucous membranes of the neck section, the chest section and the abdomen section of the esophagus, the suspicious lesion area 5 is shot by observation and recording, and the picture of the suspicious lesion area is scored.
Experimental groups: after the white light gastrointestinal endoscope examination is finished, the same experimental dog inserts a plastic catheter with an opening into an esophageal cavity through a gastrointestinal endoscope biopsy hole, 10ml of acetic acid with the mass concentration of 1.5% is sprayed forwards and backwards to enable the acetic acid to be uniformly distributed on the surface of a mucous membrane, the rest dyeing solution is sucked out after 1min, 10ml of toluidine blue dyeing solution with the mass concentration of 0.2% is sprayed forwards and backwards to enable the toluidine blue to be uniformly distributed on the surface of the mucous membrane, the dyeing solution is sucked out after 3min, and the rest dyeing solution and liquid in the esophageal cavity are fully sucked out after the mucous membrane is repeatedly washed and dyed by water for 3-4 times. After the washing, visit the intestines and stomach mirror and observe the mucous membrane of esophagus neck section, chest section and abdomen section in the dog esophagus, observe and take 5 departments of suspicious lesion area with the record, grade suspicious lesion area picture.
(2) Toluidine blue-acetic acid complex staining group experiment
Experiments were carried out by performing enteroscopy staining experiments on 6 beagle dogs successfully established with an esophagitis model, which were numbered TB-AA1, TB-AA2, TB-AA3, TB-AA4, TB-AA5 and TB-AA6, respectively.
Control group: the method is used for carrying out complex staining on an experimental control group with acetic acid-toluidine blue.
Experimental groups: after the white light gastrointestinal endoscope examination is finished, the same experimental dog inserts a plastic catheter with an opening into an esophageal cavity through a gastrointestinal endoscope biopsy hole, 10ml of toluidine blue dye solution with the mass concentration of 0.2% is sprayed from front to back to enable the toluidine blue to be uniformly distributed on the surface of a mucous membrane, the dye solution is sucked out after 3min, and the dyed mucous membrane is repeatedly washed by water for 3-4 times to enable the residual dye solution and liquid in the esophageal cavity to be fully sucked out. After the washing is finished, 10ml of acetic acid with the mass concentration of 1.5% is sprayed from front to back to ensure that the acetic acid is uniformly distributed on the surface of the mucous membrane, and the residual staining solution is sucked out after 1 min. The gastrointestinal endoscope is inserted into the esophagus of the dog to observe mucous membranes of the neck section, the chest section and the abdomen section of the esophagus, the suspicious lesion area 5 is observed and recorded to be shot, and the picture of the suspicious lesion area is scored.
(3) Mixed acetic acid toluidine blue staining group experiment
Experiment results of enterogastric staining experiments of 6 beagle dogs successfully established with an esophagitis model are numbered as MIX1, MIX2, MIX3, MIX4, MIX5 and MIX 6.
Control group: the method is the same as that of an acetic acid-toluidine blue complex staining group experiment control group.
Experimental groups: after the white light gastrointestinal endoscopy examination is finished, the same experimental dog inserts a plastic catheter with an opening into an esophageal cavity through a gastrointestinal endoscopy biopsy hole, 10ml of mixed toluidine acetate blue dye solution prepared in application example 1 is sprayed from front to back, so that the toluidine blue is uniformly distributed on the surface of the mucous membrane, the dye solution is sucked out after 2min, and the dyed mucous membrane is repeatedly washed by water for 3-4 times so that the residual dye solution and liquid in the esophageal cavity are fully sucked out. After the washing, visit the intestines and stomach mirror and observe the mucous membrane of esophagus neck section, chest section and abdomen section in the dog esophagus, observe and take 5 departments of suspicious lesion area with the record, grade suspicious lesion area picture.
3. Results and analysis of the experiments
1. The results of the acetic acid staining group are shown in tables 1 and 2.
TABLE 1 comparison of the confirmed diagnosis ratios of esophagitis in acetic acid gastrointestinal mirror staining experiments
As can be seen from Table 1, the rate of diagnosis of esophagitis in dogs by the white-light enteroscope is 44.44%, the rate of diagnosis of esophagitis in dogs by the 1.5% acetic acid solution-stained enteroscope is 50.00%, and the rate of diagnosis of esophagitis in dogs by the 1.5% acetic acid-stained enteroscope is 5.56% higher than that of the white-light enteroscope, which is partially improved.
TABLE 2 comparison of scores for canine esophagitis in acetic acid gastrointestinal staining experiments
As can be seen from table 2, P for the 1.5% acetic acid stained enteroscopy compared to the white light enteroscopy score was <0.05 with significant differences. The sensitivity of the 1.5% acetic acid staining enteroscope to the canine esophagitis diagnosis is 52.50%, the sensitivity of the white light enteroscope to the canine esophagitis diagnosis is 50.33%, and the sensitivity of the 1.5% acetic acid staining enteroscope to the canine esophagitis is 2.17% higher than that of the white light enteroscope, so that the sensitivity is improved. Therefore, the 1.5% acetic acid enteroscope has better clinical significance for diagnosing the canine esophagitis than the white light enteroscope.
It can be seen directly from fig. 1 that the solid line is significantly higher than the dotted line, but the two broken lines are slightly different and almost parallel, indicating that the acetic acid enteroscopy staining is more sensitive to the diagnosis of canine esophagitis than the white light gastroscope, but slightly different.
As can be seen from fig. 2, the damaged part of the esophagus under the white light microscope is blurred and is difficult to distinguish from the surrounding normal esophageal mucosa; acetic acid staining As seen in FIG. 3, the whitening of the esophageal mucosa is evident, and the damaged part of the esophagus protrudes from the surface of the esophageal mucosa, which is clearly contrasted with the surrounding normal esophageal mucosa. After 1.5% acetic acid staining, the esophageal mucosa bulges more obviously, and the damaged part bulges more than under a white light gastrointestinal endoscope. 1.5% acetic acid makes the color of the canine esophageal mucosa reddish and deepen after whitening compared with that before undyed, the esophageal mucosa structure is prominent, and the esophageal mucosa is smoothly changed from before undyed to after dyed with different heights. The damaged esophageal mucosa of the dog is redder than the normal esophageal mucosa under the white light gastrointestinal endoscopy, but the phenomenon that the overall reddish esophageal mucosa of the dog is observed under the whitening condition under the 1.5% acetic acid staining gastrointestinal endoscopy, so that the damaged esophageal mucosa and the normal esophageal mucosa are difficult to distinguish.
2. The results of the toluidine blue staining group are shown in tables 3 and 4.
TABLE 3 comparison of the ratio of definite diagnosis of esophagitis in dog by toluidine blue gastrointestinal mirror staining experiment
As can be seen from Table 3, the rate of diagnosis of canine esophagitis by white-light gastroscope is 50.00%, the rate of diagnosis of canine esophagitis by toluidine blue-stained gastroscope is 72.22%, and the rate of diagnosis of canine esophagitis by toluidine blue-stained gastroenteroscope is 22.22% higher than that by white-light gastroscope, which is significantly improved.
TABLE 4 comparison of canine esophagitis scores in toluidine blue gastrointestinal staining experiments
As can be seen from table 4, P for toluidine blue stained enteroscopy compared to white light enteroscopy score was <0.05 with significant differences. The sensitivity of the toluidine blue-stained gastrointestinal endoscope to the canine esophagitis diagnosis is 83.00%, the sensitivity of the white-light gastrointestinal endoscope to the canine esophagitis diagnosis is 55.17%, and the sensitivity of the toluidine blue-stained gastrointestinal endoscope to the canine esophagitis is 27.83% higher than that of the white-light gastrointestinal endoscope, so that the toluidine blue-stained gastrointestinal endoscope has obvious improvement. Therefore, the toluidine blue stained enteroscope has better clinical significance for diagnosing the canine esophagitis than the white enteroscope.
As can be seen from FIG. 4, the solid-line toluidine blue-stained gastrointestinal mirror group is significantly higher than the dotted-line white-light gastrointestinal mirror group, and the difference of the two groups of diagnostic sensitivities is obviously visually seen, which indicates that the toluidine blue-stained gastrointestinal mirror is more effective in diagnosing canine esophagitis than the white-light gastrointestinal mirror.
As can be seen from fig. 5 and 6, by comparing the pictures before and after the canine esophageal mucosa staining, the esophageal mucosa structure is more obvious after the toluidine blue staining. When the esophageal mucosa is inflamed, the pathological esophageal mucosa is not obviously displayed under the white light mirror, when a light source of the probe irradiates the esophageal mucosa, the esophageal mucosa is reddish due to light, and the normal esophageal mucosa is easily mistaken for the pathological esophageal mucosa, so that the diagnosis of the esophagitis under the white light mirror is more difficult. After the canine esophageal mucosa is observed after the canine esophagitis is dyed by toluidine blue under a gastrointestinal endoscope, the damaged esophageal mucosa can be obviously dyed to be dark blue and is shown with jewel blue dense dots, while the normal esophageal mucosa is only dyed to be light blue or not dyed, so that the damaged esophageal mucosa and the normal esophageal mucosa form a sharp contrast, and great help is provided for the diagnosis of the canine esophagitis.
3. The results of the complex staining set with acetic acid-toluidine blue are shown in tables 5 and 6.
TABLE 5 comparison of the number of confirmed diagnoses of acetic acid-toluidine blue complex staining experiment dog for esophagitis
As can be seen from Table 5, the rate of diagnosis of canine esophagitis by white-light enteroscope is 55.56%, the rate of diagnosis of canine esophagitis by toluidine blue complex-stained enteroscope is 72.22%, and the rate of diagnosis of canine esophagitis by toluidine blue-stained enteroscope is 16.66% higher than that by white-light enteroscope, which is remarkably improved.
TABLE 6 comparison of canine esophagitis scores for complex acetic acid-toluidine blue staining
As can be seen from FIG. 7, the boundary between the injured mucous membrane and the normal mucous membrane is not clear, and the color of the mucous membrane is greatly different; as can be seen from fig. 8, the damaged mucosa shows a dark blue color with uniform color tone, and the damaged mucosa is clearly demarcated from the normal mucosa to form a distinct boundary, so that the damaged mucosa is prominently protruded.
4. The results of the toluidine blue-acetic acid complex staining group are shown in table 7 and table 8.
TABLE 7 comparison of the number of confirmed diagnoses of dog esophagitis stained with toluidine blue-acetic acid
As can be seen from Table 7, the rate of diagnosis of canine esophagitis by white-light enteroscope is 55.56%, the rate of diagnosis of canine esophagitis by toluidine blue-acetic acid complex staining enteroscope is 83.33%, and the rate of diagnosis of canine esophagitis by toluidine blue-acetic acid complex staining enteroscope is 27.77% higher than that by white-light enteroscope, which is remarkably improved.
TABLE 8 comparison of toluidine blue-acetic acid stained canine esophagitis scores
As can be seen from table 8, P of toluidine blue-acetic acid complex stained enteroscopy compared with white light enteroscopy score is less than 0.05, with significant difference. The sensitivity of the toluidine blue-acetic acid compound staining enteroscope to the canine esophagitis diagnosis is 85.00 percent, the sensitivity of the white light enteroscope to the canine esophagitis diagnosis is 53.67 percent, and the sensitivity of the toluidine blue-acetic acid compound staining enteroscope to the canine esophagitis is 31.33 percent higher than that of the white light enteroscope, so that the toluidine blue-acetic acid compound staining enteroscope has obvious improvement. Therefore, the toluidine blue-acetic acid compound staining enteroscope has better clinical significance for diagnosing the canine esophagitis than a white light enteroscope.
It can be seen from fig. 9 that the boundary between the damaged mucosa and the normal mucosa is not clear, and the color of the mucosa is different, and it can be seen from fig. 10 that the damaged mucosa is displayed as blue with uniform color, the boundary between the damaged mucosa and the normal mucosa is clear, an obvious boundary is formed, and the damaged mucosa is protruded obviously.
5. The results of the mixed toluidine acetate blue staining groups are shown in tables 9 and 10.
TABLE 9 comparison of confirmed diagnosis numbers of canine esophagitis in the mixed toluidine acetate blue staining group
As can be seen from table 9, the diagnosis rate of the white-light enteroscope for canine esophagitis is 55.56%, the diagnosis rate of the mixed acetic toluidine blue complex-dyed enteroscope for canine esophagitis is 94.44%, and the diagnosis rate of the mixed acetic toluidine blue complex-dyed enteroscope for canine esophagitis is 38.88% higher than that of the white-light enteroscope, which is significantly improved.
TABLE 10 comparison of canine esophagitis scores for mixed toluidine acetate blue stained groups
P <0.05 compared to the white light enteroscopy score for mixed toluidine acetate blue stained enteroscopy from table 10 is significantly different. The sensitivity of the mixed toluidine acetate blue-stained gastrointestinal endoscope to the canine esophagitis diagnosis is 92.10%, the sensitivity of the white-light gastrointestinal endoscope to the canine esophagitis diagnosis is 56.00%, and the sensitivity of the mixed toluidine acetate blue-stained gastrointestinal endoscope to the canine esophagitis is 36.17% higher than that of the white-light gastrointestinal endoscope, so that the sensitivity is obviously improved. Therefore, the mixed acetic toluidine blue staining enteroscope has better clinical significance for diagnosing the canine esophagitis than the white light enteroscope.
It can be seen from fig. 11 that the damaged mucosa appears pink-white with uneven hue, and from fig. 12 it can be seen that the damaged mucosa appears blue with dense sapphire blue dots.
The six groups of staining experiments are summarized and counted, the experimental results can be intuitively analyzed through a chart, the result analysis is shown in fig. 13, in a single staining experiment, the diagnosis rate of the canine esophagitis by the enteroscope is ranked from high to low as comparative example 2 (toluidine blue staining group), comparative example 1 (acetic acid staining group), white light enteroscope and comparative example 3 (Lugol iodine liquid staining group), and therefore, the Lugol iodine liquid enteroscope staining does not indicate the diagnosis rate of the canine esophagitis by the enteroscope. In a single staining experiment, the enterogastroscopic staining with 1% Lugol iodine solution did not help the diagnosis of canine esophagitis, but decreased the rate of certainty in canine esophagitis. In the complex staining experiment, the diagnosis rates of canine esophagitis by gastrointestinal endoscopy were ranked from high to low as application example 1 (mixed toluidine acetate blue staining group), comparative example 5 (toluidine blue-acetic acid complex staining group), and comparative example 4 (acetic acid-toluidine blue complex staining group).
As can be seen from fig. 14, the rates of improvement of the rate of confirmed diagnosis of canine esophagitis by the stained enteroscope compared to the white-light enteroscope were ranked from high to low as application example 1 (mixed toluidine blue staining group), comparative example 5 (toluidine blue-acetic acid complex staining group), comparative example 2 (toluidine blue staining group), comparative example 4 (acetic acid-toluidine blue complex staining group), comparative example 1 (acetic acid staining group), and comparative example 3 (lugol's iodine solution staining group).
As can be seen from fig. 15, in the single staining experiment, the susceptibility of the enteroscope to canine esophagitis was ranked from high to low as comparative example 2 (toluidine blue staining group), comparative example 1 (acetic acid staining group), white light enteroscope, comparative example 3 (lugol's iodine staining group), and the 1% lugol's iodine enteroscope staining did not help in the diagnosis of canine esophagitis and also reduced the susceptibility to canine esophagitis. In the complex staining experiment, the sensitivity of the stained gastrointestinal endoscope to canine esophagitis was ranked from high to low as application example 1 (mixed toluidine acetate blue staining group), comparative example 5 (toluidine blue-acetic acid complex staining group), and comparative example 4 (acetic acid-toluidine blue complex staining group).
As can be seen visually in fig. 16, the rates of the improvement in the canine esophagitis sensitivity of the dyed enteroscopes compared to the white-light enteroscopes were ranked from high to low as application example 1 (mixed toluidine blue staining set), comparative example 5 (toluidine blue-acetic acid complex staining set), comparative example 2 (toluidine blue staining set), comparative example 4 (acetic acid-toluidine blue complex staining set), comparative example 1 (acetic acid staining set), and comparative example 3 (lugol's iodine fluid staining set).
As can be seen, the staining method of application example 1 (mixed acetic toluidine blue staining group) has higher diagnosis rate and sensitivity to canine esophagitis.
The invention judges the effect of the gastrointestinal endoscope staining mode by comparing each gastrointestinal endoscope staining mode with the change of the diagnosis rate and sensitivity under the white light gastrointestinal endoscope. In order to ensure the accuracy of the experiment, the control group and the experimental group of each gastrointestinal endoscopy staining mode are carried out in the same beagle dog, and meanwhile, in order to avoid errors generated in time, pictures and videos shot by the control group and the experimental group are arranged in the same gastrointestinal endoscopy examination, so that the control group and the experimental group are ensured to be completed on the same experimental object as much as possible, and the accuracy of the experiment is improved. In addition, six different beagle dogs are selected for each gastrointestinal mirror staining mode to carry out repeated experiments, so that the generation of experimental errors is reduced. The dyeing effect of each gastrointestinal endoscope is determined by comparing the dyeing mode with the white-light gastrointestinal endoscope of the group, so that the error of different diagnosis rates and sensitivities of the white-light gastrointestinal endoscopes among different groups is avoided, and the experimental result has higher reliability.
The diagnosis rate of the staining method of comparative example 1 (acetic acid staining group) on the canine esophagitis is 50.00%, the sensitivity is 52.50%, the improvement rate of the staining method compared with the white light enterogastroscope examination of the same group is 12.51%, the improvement rate of the sensitivity is 4.31%, the diagnosis assistance on the canine esophagitis is not large, and the diagnosis is related to that the acetic acid is weak acid and has certain irritation on the canine esophagitis mucosa, although 1.5% acetic acid can cause a transient acetowhitening reaction on the canine esophagitis mucosa, and the acidic irritation of 1.5% acetic acid causes swelling and reddening of the canine esophagitis mucosa, so that the acetic acid has poor whitening effect on the canine esophagitis mucosa. After 1.5% acetic acid is dyed, normal esophagus and damaged esophagus of a dog are pink after whitening, so that the damaged esophageal mucosa is difficult to distinguish from the normal esophageal mucosa, and the acetic acid only plays a role in highlighting the esophageal mucosa structure in dyeing the esophageal mucosa of the dog, so that the damaged esophageal mucosa is more prominent than the normal esophageal mucosa and is easier to find.
The staining method of comparative example 2 (toluidine blue staining group) has the diagnosis rate of 72.22% and the sensitivity of 83.00% for canine esophagitis, and the improvement rate of 44.44% compared with the white light enterogastroscopy diagnosis rate of the same group, the sensitivity of 50.44% is improved, cations in toluidine blue have staining effect, have strong affinity with acid groups (including carboxylic acid, sulfuric acid, phosphate and the like) of DNA and RNA in cells, can stain cell nuclei to be blue, and the staining degree of cells can reflect the content of DNA and RNA in cells. Toluidine blue also has a strong affinity for mitotically active cells, such as abnormally proliferating epithelial cells, and therefore enables staining thereof.
The diagnosis rate of the staining method of comparative example 3 (Lugol's iodine solution staining group)) for canine esophagitis was 38.83%, the sensitivity was 52.17%, the improvement rate of the diagnosis rate was-12.62% and the improvement rate of the sensitivity was-1.57% compared to the white light gastrointestinal examination of the same group, and thus it was found that a single Lugol's iodine solution gastrointestinal staining was not suitable for canine esophagitis diagnosis. Lugol's iodine solution is dark brown staining solution, which is mainly used in human medicine to screen esophageal cancer, and cancer cells are rapidly replicated and divided, so that glycogen supply in a cancerous region is insufficient, and no sugar in the cancerous region reacts with Lugol's iodine solution to form glycogen reaction, so that the cancerous region is in a non-staining state. However, although glycogen is consumed in the area with inflammation caused by esophageal injury, glycogen is not consumed as much as cancer tissues, so the inflammation area of the canine esophageal mucosa still reacts with Lugol iodine solution to form glycogen reaction, and the staining of the injury area is lighter. Although the staining of the damaged esophageal mucosa area of the dog is light, the damaged esophageal mucosa area of the dog is not easily distinguished from the brown color of the normal esophageal mucosa, so that the diagnosis effect on the canine esophagitis is reduced.
The diagnosis rate of canine esophagitis by the staining method of comparative example 4 (acetic acid-toluidine blue complex staining group) is 72.22%, and the improvement rate is 29.99% compared with a white light gastroscope; the sensitivity was 81.67%, and the lift ratio was 43.70% compared to a white gastroscope. The dyeing effect of the dyeing method of the comparative example 2 (toluidine blue dyeing group) is better than that of the comparative example 4 (acetic acid-toluidine blue composite dyeing group), because 10ml of 1.5% acetic acid is firstly used for dyeing canine esophageal mucosa, so that esophageal mucosa is swollen, squamous epithelial cells of the esophageal mucosa are more closely arranged, 0.2% toluidine blue dye solution is difficult to enter intercellular spaces, so that the dye solution is difficult to be absorbed by cells, and the dyeing effect of the 0.2% toluidine blue dye solution on the canine esophageal mucosa is reduced.
The diagnosis rate of the staining method of comparative example 5 (toluidine blue-acetic acid composite staining group) on the canine esophagitis is 83.33%, the sensitivity is 85.00%, and compared with the white light gastrointestinal endoscopy examination of the same group, the diagnosis rate is 49.98%, and the sensitivity is improved by 58.38%. Compared with the comparative example 2 (toluidine blue staining group), the diagnosis rate of the dog esophagitis diagnosis of the comparative example 5 (toluidine blue-acetic acid composite staining group) is improved by 5.54 percent, and the sensitivity is improved by 7.94 percent. The canine esophageal mucosa is firstly dyed with toluidine blue and then acetic acid is used, the whitening effect of the acetic acid does not reduce the dyeing effect of the toluidine blue, the acetic acid enables the esophageal mucosa to swell and be transparent, the damaged esophageal mucosa dyed with blue is more prominent, and the dyeing effect of the esophageal mucosa is improved.
The diagnosis rate of dog esophagitis by using the staining method of example 1 (mixed toluidine acetate blue staining group) is 94.44%, the sensitivity is 92.17%, and compared with the white light enteroscopy diagnosis rate of the same group, the diagnosis rate is increased by 69.99%, and the sensitivity is increased by 64.59%. The acetic acid is added into toluidine blue staining solution to ensure that the staining effect is better, because the acetic acid causes the canine esophageal mucosa to swell, the squamous epithelial cells of the esophageal mucosa become bigger, the permeability of cell membranes is increased, and meanwhile, toluidine blue is easier to be absorbed by cells to stain the cells.
Acetic acid in the acetic acid-toluidine blue compound dyeing group is not beneficial to the absorption of toluidine blue dye liquor, but acetic acid in the mixed acetic acid toluidine blue dyeing group is more beneficial to the absorption of toluidine blue, and different effects are caused because the time nodes of the acetic acid in the esophageal mucosa hair dyeing effect are different. In the acetic acid-toluidine blue composite staining group, acetic acid is firstly used for staining, after esophageal mucosa cells swell, toluidine blue is used for staining the swollen esophageal mucosa, at the moment, the esophageal mucosa cells are closely arranged, and the added toluidine blue staining solution is difficult to enter the intercellular spaces and is difficult to absorb, so that the staining effect of the staining group is poorer than that of the mixed acetic acid toluidine blue staining group. In the mixed acetic acid toluidine blue staining group, acetic acid and toluidine blue are mixed and then simultaneously act on esophageal mucosa, the acetic acid enables the esophageal mucosa to swell, and toluidine blue in the mixed solution is easier to enter cells and combine with intracellular DNA in the process of increasing the permeability of the cell membrane in the mixed solution, so that the toluidine blue staining effect is better. Meanwhile, acetic acid enables the morphological gradation of the esophageal mucosa to be clearer, and the dyeing effect of toluidine blue is improved.
The canine esophageal mucosa staining method is used for staining canine esophageal mucosa by the staining methods of application example 1, application example 2 and application example 3, the diagnosis rate and sensitivity results are not obviously different, and the method takes application example 1 as an example and is compared with other comparative examples in experiments.
The above description is only a preferred embodiment of the present application and is not intended to limit the present application, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present application shall be included in the protection scope of the present application.
Claims (9)
1. A composite coloring agent is characterized in that: the composite coloring agent is a mixed solution consisting of an acetic acid solution, a toluidine blue solution and normal saline, wherein the mass concentration of acetic acid in the mixed solution is 1.5-2%, and the mass concentration of toluidine blue in the mixed solution is 0.2-0.4%.
2. The composite coloring agent according to claim 1, characterized in that: the mass concentration of acetic acid in the mixed solution was 1.5%, and the mass concentration of toluidine blue was 0.2%.
3. The method for preparing a composite coloring agent according to any one of claims 1 to 2, characterized by comprising the steps of: mixing acetic acid solution, toluidine blue solution and normal saline to obtain the composite coloring agent.
4. The production method according to claim 3, characterized in that: in the preparation of the composite coloring agent, the mass concentration of the adopted acetic acid solution is 3-4%, and the mass concentration of the toluidine blue solution is 1-2%.
5. The production method according to claim 3, characterized in that: the volume ratio of acetic acid solution, toluidine blue solution and normal saline is 5:2:3 when the composite coloring agent is prepared.
6. The use of the composite coloring agent of claim 1 in the preparation of a staining reagent for canine esophageal mucosa.
7. The use of the composite coloring agent of claim 1 for preparing a diagnostic reagent for canine esophagitis.
8. The method for staining the esophageal mucosa of the dog is characterized by comprising the following steps: spraying the composite coloring agent of any one of claims 1-2 into the esophagus of a dog, sucking out the residual composite coloring agent in the esophagus and flushing the esophagus after 2-3 min.
9. The method of claim 8, wherein: the volume of the composite coloring agent sprayed each time is 10-15 ml.
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| CN115414323A (en) * | 2022-08-25 | 2022-12-02 | 遵义医科大学 | General anesthesia method for experimental pigs |
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| CN112985953A (en) * | 2021-02-22 | 2021-06-18 | 武汉大学 | Surgical specimen marking method using magnifier and dye |
| CN115414323A (en) * | 2022-08-25 | 2022-12-02 | 遵义医科大学 | General anesthesia method for experimental pigs |
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