CN111398276A - Evaluation method of fish intestinal tissue slices - Google Patents
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Abstract
The invention discloses an evaluation method of fish intestinal tissue slices, which belongs to the field of aquatic products and comprises the following steps: observing the mucosa layer, the submucosa layer, the muscular layer and the serous layer of the target tissue section by using a microscope, and respectively scoring the observation symptom results; the weights of mucosa, submucosa, muscularis and serosa are b1, b2, b3 and b4 respectively, wherein b1> b2 and b3> b4, and b1+ b2+ b3+ b4 is 1; scoring of intestinal tissue sections. The method adopts a weighted scoring method to establish an index scoring standard of pathological symptoms of the intestinal tissues, carries out comprehensive quantitative evaluation on the intestinal tissue slices, determines the weights of different symptoms of the same structural layer and the weights of different structural layers of the intestinal tissues according to the importance degree, finally multiplies the grade scores of the symptoms by the symptom weights to obtain the scores of the structural layers, comprehensively considers all symptoms of each intestinal slice, finally converts the symptoms into data for statistical analysis, and has objective and comprehensive data.
Description
Technical Field
The invention relates to the field of aquatic products, in particular to an evaluation method of fish intestinal tissue slices.
Background
The currently used evaluation method of intestinal slices is to represent the level of the treatment group by description or measurement of some local parts of an individual intestinal slice. However, the difference of intestinal tissue sections is large, mainly manifested by: the intestinal sections of different fishes in the same treatment group are different, the pathological changes of different tissue structures in the same intestinal ring are different, and the different visual fields of the same tissue structure in the same intestinal ring are different. Secondly, the quantifiable indexes of the intestinal slices are not many, and the measurement result is easily influenced by the quality of the slices and the subjectivity of an observer. For example, to measure pile height, the piles are not uniform in size, are bent or folded, and the observer often selects the longest 10 piles to measure and take the average. It can be seen that some local descriptions or measurements of a slice are difficult to represent the overall level of the treatment group and are clearly one-sided and subjective.
Disclosure of Invention
In order to solve the problems of subjective one-sided evaluation results, large difference, low accuracy of evaluation structure and the like in the prior art, the invention provides the evaluation method of the fish intestinal tissue slices, forms a set of relatively systematic, comprehensive and complete evaluation method, and improves the systematicness and accuracy of the fish intestinal tissue slice evaluation.
The specific technical scheme is as follows:
a method for evaluating fish intestinal tissue slices comprises the following steps:
the method comprises the following steps: observing the mucosa layer, the submucosa layer, the muscular layer and the serous layer of the target tissue section by using a microscope, and scoring the observation symptom results respectively, wherein the scores of the four structural layers are respectively c1, c2, c3 and c4, the scoring range is 0-M, M is a certain integer from 1 to 100, and the larger the numerical value from 0-M is, the more serious the pathological change of the corresponding structural layer is; the weights of mucosa, submucosa, muscularis and serosa are b1, b2, b3 and b4 respectively, wherein b1> b2 and b3> b4, and b1+ b2+ b3+ b4 is 1;
step two: scoring of intestinal tissue sections
The score D ranged from 0-M, with larger values indicating more severe lesions in the intestinal tissue sections.
Preferably, the scores of the mucosa layer, the submucosa layer, the muscularis layer and the serosal layer in the intestinal tissue section adopt a weighted scoring method, pathological symptoms in the four structural layers are listed one by one, each pathological symptom score A is divided into 6 grades, and the scores are respectively 0,
M, wherein 0 represents normal;
indicating occasional mild changes;
indicates a mild change;
indicating a moderate change;
indicating a severe change; "M" means complete change, complete regression of function, villous necrosis, or extreme edema.
Preferably, the mucosa pathology comprises 10 pathologies, each pathology having a score aa1-Aa10The weight of each pathology in the mucosal layer is a1-a10, respectively, and the correspondence is as follows:
a1 is the weight of villus branch, a2 is the weight of villus fusion and necrosis, a3 is the weight of villus fracture disintegration or damage, a4 is the weight of villus edema, a5 is the weight of villus inflammatory cell infiltration, a6 is the weight of villus hemorrhage, a7 is the weight of sparse villus, a8 is the weight of villus disorganization, a9 is the weight of villus shortness, a10 is the weight of villus deletion, and a1+ a2+ a3+ a4+ a5+ a6+ a7+ a8+ a9+ a10 ═ 1;
scoring of the mucosal layer
Preferably, the submucosal pathology comprises 4 pathologies, each pathology scored a respectivelya11-Aa14The weight of each pathology in the mucosal layer is a11-a14, respectively, and the correspondence is as follows:
a11 is the weight of submucosa loss, a12 is the weight of submucosa edema, a13 is the weight of submucosa hemorrhage, a14 is the weight of submucosa separation from muscularis, and a11+ a12+ a13+ a14 is 1;
scoring of the submucosa
Preferably, the muscle layer pathology comprises 5 pathologies, each pathology having a score aa15-Aa19The weight of each pathology in the muscle layer is a15-a19, and the corresponding relation is as follows:
a15 is the weight of inner ring muscle loss, a16 is the weight of inner ring muscle edema, a17 is the weight of inner ring muscle bleeding, a18 is the weight of outer longitudinal muscle loss, a19 is the weight of outer longitudinal muscle edema, and a15+ a16+ a17+ a18+ a19 is 1;
scoring of the muscle layer
Preferably, the serosal pathology comprises 4 pathologies, each pathology scoring a respectivelya20-Aa23The weight of each pathology in the serosal layer is a20-a23, respectively, and the corresponding relationship is as follows:
a19 is the weight of serosal layer damage, defect, a20 is the weight of serosal layer edema, a21 is the weight of serosal layer hemorrhage, a22 is the weight of lateral longitudinal muscle loss, a23 is the weight of serosal layer-muscle layer separation, and a19+ a20+ a21+ a22+ a23 is 1;
scoring of the serosal layer
Preferably, the weights b1, b2, b3 and b4 of the mucosa layer, the submucosa layer, the muscle layer and the serosa layer are respectively 0.5, 0.2 and 0.1.
Preferably, M has a value of 40.
Preferably, the weights a1-a10 are 0.06, 0.25, 0.12, 0.02, 0.32, respectively.
Preferably, the weights a11-a14 are 0.3, 0.1, respectively.
Preferably, the weights a15-19 have values of 0.3, 0.2, 0.1, respectively.
Preferably, the weights a20-23 have values of 0.3, 0.1, respectively.
The beneficial effects of the evaluation method provided by the invention are as follows:
and establishing a grading standard of the pathological symptom indexes of the intestinal tissues by adopting a weighted grading method, and carrying out comprehensive quantitative evaluation on the intestinal tissue slices. All pathological symptoms observed in the four-layer structure of the intestinal tissue (mucosa layer, submucosa layer, muscle layer and serosa layer) are listed one by one, and each pathological symptom is divided into 6 grades and scores. And determining the weights of different symptoms of the same structural layer and the weights of different structural layers of intestinal tissues according to the importance degree. Finally, multiplying the grade scores of all symptoms by the symptom weights to obtain the scores of all structural layers; and multiplying the score of each structural layer by the weight of the structural layer to obtain the total score of the intestinal slice, wherein the higher the score is, the more serious the intestinal pathological symptoms are. The method comprehensively considers all symptoms of each intestinal section, and finally converts the symptoms into data for statistical analysis, and the data are objective and comprehensive.
Drawings
FIG. 1 is a microscopic section of the posterior intestine of a preferred embodiment of the present invention (HE staining, 50 ×);
FIG. 2 is a photograph of a preferred microscopic posterior section of a moderate lesion of the present invention (HE staining, 50 ×);
FIG. 3 is a photograph of a microscopically lightly lesioned hindgut section (HE stain, 40 ×) in accordance with a preferred embodiment of the present invention;
FIG. 4 is a microscopic view of a normal section of the hindgut of a preferred embodiment of the present invention (HE stain, 100 ×).
Detailed Description
The present invention will be described in further detail with reference to examples. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The beneficial effects of the evaluation method provided by the invention are as follows: and establishing a grading standard of the pathological symptom indexes of the intestinal tissues by adopting a weighted grading method, and carrying out comprehensive quantitative evaluation on the intestinal tissue slices. All pathological symptoms observed in the four-layer structure of the intestinal tissue (mucosa layer, submucosa layer, muscle layer and serosa layer) are listed one by one, and each pathological symptom is divided into 6 grades and scores. And determining the weights of different symptoms of the same structural layer and the weights of different structural layers of intestinal tissues according to the importance degree. Finally, multiplying the grade scores of all symptoms by the symptom weights to obtain the scores of all structural layers; and multiplying the score of each structural layer by the weight of the structural layer to obtain the total score of the intestinal slice, wherein the higher the score is, the more serious the intestinal pathological symptoms are. The method comprehensively considers all symptoms of each intestinal section, and finally converts the symptoms into data for statistical analysis, and the data are objective and comprehensive. The scoring criteria of the pathological symptom indexes of the intestinal tissues are shown in the table 1.
TABLE 1 Scoring standard for pathological symptom index of intestinal tissue
The above table is specifically illustrated as follows:
(1) organization structure: the tissue structures of the front, middle and back intestines of the fish are respectively a mucous layer, a submucosa layer, a muscular layer and a serosal layer from the intestinal cavity to the intestinal wall in sequence. Wherein the mucosal layer is composed of intestinal epithelial cells and lamina propria which bulge toward the intestinal lumen to form a plurality of finger-like projections called intestinal villi. The submucosa consists of loose connective tissue and is the strut of the mucosal layer. The muscle layer is composed of smooth muscle, the inner layer is the ring muscle, and the outer layer is the longitudinal muscle. The serosal layer consists of a thin layer of loose joint tissue that is associated with the mesentery by an outer layer of epithelial cells extending from the mesentery.
(2) Grading and scoring of symptoms: the mucosa, submucosa, muscularis and serosa have corresponding symptoms. For example, the mucosal layer has 10 symptoms such as villus branch … … villus loss, the submucosa has 4 symptoms such as submucosa … … submucosa separated from the muscularis, the muscularis has 5 symptoms such as inner ring muscle loss … … outer longitudinal muscle edema, and the serosal layer has 4 symptoms such as serosal layer damage, and defect … … serosal layer separated from the muscularis. Each symptom is divided into 5 to 6 grades, each grade is represented by different symbols, wherein, 0 represents normal; "±" indicates occasional mild changes; "+" indicates a slight change; "+ +" indicates a moderate change; "+ + + +" indicates severe change; "+ ++" indicates complete change, complete regression of function, villous necrosis, or extreme edema. The scores of the grades are 0, 5, 10, 15, 30 and 40 in sequence, and the score is higher as the symptom is worse. In each symptom, the villus branch is beneficial to enlarging the digestive absorption area of the intestinal tract, belongs to a beneficial index, and the scores of all grades are 0, -5, -10, -15 and-30 in sequence.
(3) Weight of different symptoms of the same structural layer:
a mucous membrane layer: intestinal villi are sites for digestion and absorption of nutrients and also are a defense barrier, and have fusion necrosis, villi fracture disintegration or damage, the greatest influence on the digestion and absorption functions of the mucosa layer, the weights of the intestinal villi on the mucosa layer are also the greatest, and are respectively 0.25, and the total weight of the intestinal villi and the mucosa layer is 0.5. The edema, infiltration of inflammatory cells and hemorrhage of the villi are also pathological changes of the villi, but the influence on the function of the mucosa is smaller than the above two symptoms, so the weight is 0.12 respectively, and the total weight of the three is 0.36.
Submucosa: submucosa loss, edema, bleeding, and a greater influence on submucosa function, the weight of submucosa is also greater, 0.3 for each, and the total of the three is 0.9.
Muscle layer: rhythmic contraction and relaxation of the inner ring muscle are the main motive force for segmental movement of the intestinal tract, and the outer longitudinal muscle is the contraction and relaxation mainly forms the swing of the intestinal tract. The influence of the inner ring muscle on the function of the muscle layer is far greater than that of the outer longitudinal muscle, and the weight of 3 symptoms of the inner ring muscle on the muscle layer is 0.8 in total; wherein the internal ring muscle loss and edema have great influence on the peristalsis of the intestinal tract and are not beneficial to the backward movement and digestion of the bolus, so the weight of the 2 symptoms in the muscle layer is also the largest and is respectively 0.3.
A serosal layer: the symptoms that greatly affect the serosa layer are serosa layer defect, edema, and hemorrhage, and the weights of the serosa layers are 0.3 and 0.9 in total.
(4) Weight of different structural layers: the function of the 4-layer structure of the intestinal tract tissue is different, and the important degrees of the intestinal tract health are as follows in sequence: mucosal layer > submucosal layer, muscular layer > serosal layer, so their weights are 0.5, 0.2, 0.1, respectively.
(5) Score calculation for individual tissue sections:
the score of each structural layer C is ∑ (symptom score A × weight a of different symptoms of the same structural layer), and the score of each structural layer is calculated respectively.
The final score D of the intestinal section is ∑ (score of each structural layer C ×, weight b of each structural layer).
The scores of all structural layers and the final scores of the intestinal slices participate in statistical analysis in a data form, so that the difference of the same structural layer among different treatment groups can be compared, the difference of the integral conditions of the intestinal tracts among different treatment groups can be compared, and the data is comprehensive and objective.
Wherein a severe lesion is found when the final score D satisfies 17< D ≦ 36: the grade of symptoms such as villus fusion or necrosis, villus fracture and disintegration or damage, villus edema and the like reaches + + + or above, and the total number of indexes with the grade of symptoms reaching + + + and above in 9 unfavorable indexes of the mucosa layer is more than 5; the number of indicators with symptom grade of +++ and above is more than 6 in total. Moderate lesions are obtained when D is more than 10 and less than or equal to 17: the level of villus fusion or necrosis is not higher than + +, and the number of indexes with symptom levels reaching or exceeding + + + is not more than 2 in 5 higher-weighted mucosal layer indexes (villus fusion or necrosis, villus fracture disintegration or damage, villus edema, villus inflammatory cell infiltration, villus hemorrhage); the grade of other symptoms is between 0 and + and; the number of indicators having symptom levels of + + and above is not more than 11 in total. Mild lesions are obtained when D is more than 6 and less than or equal to 10: the grade of villus fusion or necrosis is not higher than + +, the grade of symptoms such as villus fracture disintegration or damage, villus edema and the like is not higher than +, the grade of symptoms such as villus hemorrhage, submucosal edema and the like is not higher than + +, and the grade of other symptoms is between 0 and + inclusive; the number of indexes with symptom grade of + and above is not more than 11 in total. Normal when D is more than 0 and less than or equal to 6: the grade of villus fusion or necrosis is not higher than +, the grade of symptoms such as villus fracture disintegration or damage, villus edema and the like is not higher than +/-and the grade of symptoms such as villus hemorrhage, submucosal hemorrhage and the like is not higher than + +, and the grade of other symptoms is 0 or +/-; the number of indexes with symptom grades of +/-and above is not more than 11 in total.
Analyzing intestinal slices with pathological changes of different degrees based on the scoring standard of the pathological symptom indexes of the intestinal tissues:
a severely diseased intestinal section is shown in fig. 1, where villus break disintegration or damage is shown +++: lesion area > 3/4; villous fusion, necrosis +++: villi are degenerated and necrotic, and the function is completely eliminated; table 2 shows the score of the section, which is 19.25.
TABLE 2 evaluation of hindgut sections of severe lesions
As shown in fig. 2, moderately diseased intestinal sections showing villous edema + +: the edema area is 1/4-1/2; villus break disintegration or damage +++: lesion area > 3/4; villous hemorrhage ++: each villus bleeds with high red cell concentration and deep blood color (bleeding area in each villus > 1/2); table 3 shows the score of the section, which is 13.15.
TABLE 3 evaluation of hindgut sections for moderate lesions
As shown in fig. 3 is a mildly diseased intestinal section showing villous fusion, necrosis + +: most of the hair is fused, but normal villi still exist, and the hair has functions; villous hemorrhage ++: each villus is bled and has a certain area (the bleeding area in each villus is 1/3-1/2); villous edema +: edema area < 1/4; table 4 shows the score of the section, which is 7.68.
TABLE 4 evaluation of hindgut sections of moderate lesions
Normal intestinal sections are shown in fig. 4, where villous fusion, necrosis +: fusing part of villi; fluff break disintegration or damage 0: no damage and no fragment; villous hemorrhage ++: each villus is bled and has a certain area (the bleeding area in each villus is 1/3-1/2); table 5 shows the score of the section, which is 5.3.
TABLE 5 evaluation of Normal hindgut sections
While the foregoing description shows and describes the preferred embodiments of the present invention, it is to be understood that the invention is not limited to the forms disclosed herein, but is not to be construed as excluding other embodiments and is capable of use in various other combinations, modifications, and environments and is capable of changes within the scope of the inventive concept as described herein, commensurate with the above teachings, or the skill or knowledge of the relevant art. And that modifications and variations may be effected by those skilled in the art without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (12)
1. A method for evaluating fish intestinal tissue slices is characterized by comprising the following steps:
the method comprises the following steps: observing the mucosa layer, the submucosa layer, the muscular layer and the serous layer of the target tissue section by using a microscope, and scoring the observation symptom results respectively, wherein the scores of the four structural layers are respectively c1, c2, c3 and c4, the scoring range is 0-M, M is a certain integer from 1 to 100, and the larger the numerical value from 0-M is, the more serious the pathological change of the corresponding structural layer is; the weights of mucosa, submucosa, muscularis and serosa are b1, b2, b3 and b4 respectively, wherein b1> b2 and b3> b4, and b1+ b2+ b3+ b4 is 1;
step two: scoring of intestinal tissue sections
The score D ranged from 0-M, with larger values indicating more severe lesions in the intestinal tissue sections.
2. The method according to claim 1, wherein the scores of the mucosal layer, the submucosal layer, the muscular layer and the serosal layer in the intestinal tissue section are weighted and scored, pathological symptoms in the four structural layers are listed one by one, each pathological symptom score A is divided into 6 grades, and the scores are respectively 0, 0 and,
M, wherein 0 represents normal;
indicating occasional mild changes;
indicates a mild change;
indicating a moderate change;
indicating a severe change; "M" means complete change, complete regression of function, villous necrosis, or extreme edema.
3. The method of claim 2, wherein the mucosal pathologies comprise 10 pathologies, each pathology being scored aa1-Aa10The weight of each pathology in the mucosal layer is a1-a10, respectively, and the correspondence is as follows:
a1 is the weight of villus branch, a2 is the weight of villus fusion and necrosis, a3 is the weight of villus fracture disintegration or damage, a4 is the weight of villus edema, a5 is the weight of villus inflammatory cell infiltration, a6 is the weight of villus hemorrhage, a7 is the weight of sparse villus, a8 is the weight of villus disorganization, a9 is the weight of villus shortness, a10 is the weight of villus deletion, and a1+ a2+ a3+ a4+ a5+ a6+ a7+ a8+ a9+ a10 ═ 1;
scoring of the mucosal layer
4. The method of claim 3, wherein the submucosal pathology comprises 4 pathologies, each pathology scored separatelyAa11-Aa14The weight of each pathology in the mucosal layer is a11-a14, respectively, and the correspondence is as follows:
a11 is the weight of submucosa loss, a12 is the weight of submucosa edema, a13 is the weight of submucosa hemorrhage, a14 is the weight of submucosa separation from muscularis, and a11+ a12+ a13+ a14 is 1;
scoring of the submucosa
5. The method of claim 4, wherein the pathology of the muscular layer comprises 5 pathologies, and the score of each pathology is Aa15-Aa19The weight of each pathology in the muscle layer is a15-a19, and the corresponding relation is as follows:
a15 is the weight of inner ring muscle loss, a16 is the weight of inner ring muscle edema, a17 is the weight of inner ring muscle bleeding, a18 is the weight of outer longitudinal muscle loss, a19 is the weight of outer longitudinal muscle edema, and a15+ a16+ a17+ a18+ a19 is 1;
scoring of the muscle layer
6. The method of claim 5, wherein the serosal pathology comprises 4 pathologies, each pathology having a score Aa20-Aa23The weight of each pathology in the serosal layer is a20-a23, respectively, and the corresponding relationship is as follows:
a19 is the weight of serosal layer damage, defect, a20 is the weight of serosal layer edema, a21 is the weight of serosal layer hemorrhage, a22 is the weight of lateral longitudinal muscle loss, a23 is the weight of serosal layer-muscle layer separation, and a19+ a20+ a21+ a22+ a23 is 1;
scoring of the serosal layer
7. The method of claim 6, wherein the weights b1, b2, b3 and b4 of mucosal layer, submucosal layer, muscular layer and serosal layer are 0.5, 0.2 and 0.1, respectively.
8. The method according to claim 7, wherein M has a value of 40.
9. The method of claim 8, wherein the weights a1-a10 are 0.06, 0.25, 0.12, 0.02, 0.32, respectively.
10. The method for evaluating a slice of intestinal tissue of a fish according to claim 9, wherein the values of the weights a11-a14 are 0.3, 0.3 and 0.1, respectively.
11. The method for evaluating a slice of intestinal tissue of a fish according to claim 10, wherein the values of the weights a15-19 are 0.3, 0.2, 0.1 and 0.1, respectively.
12. The method according to claim 11, wherein the values of the weights a20-23 are 0.3, and 0.1, respectively.
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