CN112557347B - Preparation and detection method of chyle particle migration model in mucus layer - Google Patents
Preparation and detection method of chyle particle migration model in mucus layer Download PDFInfo
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- 210000003097 mucus Anatomy 0.000 title claims abstract description 70
- 210000001268 chyle Anatomy 0.000 title claims abstract description 49
- 230000005012 migration Effects 0.000 title claims abstract description 33
- 238000013508 migration Methods 0.000 title claims abstract description 33
- 238000001514 detection method Methods 0.000 title claims abstract description 27
- 239000002245 particle Substances 0.000 title claims abstract description 24
- 238000002360 preparation method Methods 0.000 title claims abstract description 17
- 230000000968 intestinal effect Effects 0.000 claims abstract description 44
- 241001465754 Metazoa Species 0.000 claims abstract description 10
- 210000000936 intestine Anatomy 0.000 claims abstract description 3
- 210000002429 large intestine Anatomy 0.000 claims description 21
- 230000010355 oscillation Effects 0.000 claims description 20
- 238000000149 argon plasma sintering Methods 0.000 claims description 19
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 12
- 230000000694 effects Effects 0.000 claims description 12
- 238000002156 mixing Methods 0.000 claims description 12
- 239000000243 solution Substances 0.000 claims description 12
- 108010004103 Chylomicrons Proteins 0.000 claims description 11
- 238000000034 method Methods 0.000 claims description 10
- 230000008569 process Effects 0.000 claims description 10
- 239000000126 substance Substances 0.000 claims description 10
- 239000004382 Amylase Substances 0.000 claims description 8
- 102000013142 Amylases Human genes 0.000 claims description 8
- 108010065511 Amylases Proteins 0.000 claims description 8
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 8
- 108010019160 Pancreatin Proteins 0.000 claims description 8
- 102000057297 Pepsin A Human genes 0.000 claims description 8
- 108090000284 Pepsin A Proteins 0.000 claims description 8
- 235000019418 amylase Nutrition 0.000 claims description 8
- 239000011259 mixed solution Substances 0.000 claims description 8
- 229940055695 pancreatin Drugs 0.000 claims description 8
- 229940111202 pepsin Drugs 0.000 claims description 8
- 238000003825 pressing Methods 0.000 claims description 6
- 238000001228 spectrum Methods 0.000 claims description 6
- 238000012360 testing method Methods 0.000 claims description 6
- 239000007853 buffer solution Substances 0.000 claims description 5
- 102000004190 Enzymes Human genes 0.000 claims description 4
- 108090000790 Enzymes Proteins 0.000 claims description 4
- 229940124158 Protease/peptidase inhibitor Drugs 0.000 claims description 4
- 229940088598 enzyme Drugs 0.000 claims description 4
- 235000011389 fruit/vegetable juice Nutrition 0.000 claims description 4
- 239000000137 peptide hydrolase inhibitor Substances 0.000 claims description 4
- 210000003296 saliva Anatomy 0.000 claims description 4
- 230000000087 stabilizing effect Effects 0.000 claims description 4
- 210000004051 gastric juice Anatomy 0.000 claims description 3
- 239000007788 liquid Substances 0.000 claims description 3
- 238000003307 slaughter Methods 0.000 claims description 3
- 238000004140 cleaning Methods 0.000 claims 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 230000006399 behavior Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 230000001079 digestive effect Effects 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 210000005027 intestinal barrier Anatomy 0.000 description 1
- 230000007358 intestinal barrier function Effects 0.000 description 1
- 210000004400 mucous membrane Anatomy 0.000 description 1
- 210000004798 organs belonging to the digestive system Anatomy 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 230000032258 transport Effects 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/47—Scattering, i.e. diffuse reflection
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/59—Transmissivity
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/47—Scattering, i.e. diffuse reflection
- G01N2021/4704—Angular selective
- G01N2021/4709—Backscatter
Abstract
The invention discloses a preparation and detection method of a chyle particle migration model in a mucus layer, which relates to the technical field of preparation and detection of particle migration models and comprises the following steps: s1, obtaining animal intestine mucus; s2, preparing artificial chyle, and preserving under certain conditions; s3, adding the intestinal mucus into a sample bottle, and then adding the artificial chyle. The preparation and detection method of the chyle particle migration model in the mucus layer solves the problems of long experimental period, high operation difficulty, ethical concern and the like of conventional animals.
Description
Technical Field
The invention relates to the technical field of preparation and detection of particle migration models, in particular to a preparation and detection method of a chyle particle migration model in a mucus layer.
Background
The intestinal tract is an important digestive organ of a human body, and the surface of the intestinal tract is covered with a layer of mucus, and epithelial cells and harmful substances such as bacteria and viruses in the intestinal tract are effectively isolated and removed through continuous secretion and renewal of the mucus, so that the intestinal tract mucous membrane is lubricated and protected. Mucus layer is an important constituent constituting the intestinal barrier. Chylomicrons play an important role in the digestive absorption process of food. Studying the migration of chylomicrons reveals the absorption and transport process of substances in intestinal mucus and interactions of chylomicrons with intestinal mucus during migration.
Animal experiments are often selected for migration of chyle particles in the mucus layer in the past, but the animal experiments have long period, are greatly influenced by environment, have higher cost and have higher professional technical requirements for experimenters. Ethical issues are also involved in dissecting animals. The external digestion model is widely applied because of the advantages of simple construction, low cost, high repeatability, capability of realizing simultaneous detection of multiple samples, no ethical limitation and the like.
The invention aims to provide a preparation and detection method of a chylomicron migration model in a mucus layer, and the prepared model is used for simply and clearly characterizing chylomicron migration.
Disclosure of Invention
(one) solving the technical problems
Aiming at the defects of the prior art, the invention provides a preparation and detection method of a chylomicron migration model in a mucus layer, which solves the problem of simply and clearly researching the chylomicron migration by simulating the mucus layer in vitro.
(II) technical scheme
In order to achieve the above purpose, the invention adopts the following technical scheme: a preparation method of a chyle particle migration model in a mucus layer comprises the following steps:
s1, obtaining animal intestinal mucus.
S2, preparing the artificial chyle, and preserving under certain conditions.
S3, adding the intestinal mucus into a sample bottle, and then adding the artificial chyle.
In the step S1, the intestinal mucus is obtained from the large intestine of a pig, the large intestine of the pig is fresh, preferably, the large intestine of the pig is taken immediately after slaughtering to extract the intestinal mucus, the intestinal mucus can be stored on ice cubes in the transportation process, the dirt on the surface of the large intestine of the pig is gently washed by using PBS buffer solution before the intestinal mucus is extracted, but the large intestine of the pig is sheared into small sections of 15-20 cm after the washing is completed, the surface of the large intestine is rubbed with force to collect the mucus, and protease inhibitor is added into the intestinal mucus for storage under the condition of-20 ℃.
In the step S2, firstly, fully dissolving a substance to be detected according to the required concentration, mixing the substance with simulated saliva according to the volume ratio of 1:1, adding salivary amylase to ensure that the enzyme activity value of the salivary amylase is 70-100U/mL, adjusting the pH value to be 7, and standing for 2-5 min at the temperature of 25 ℃;
mixing the mixed solution and simulated gastric juice according to the volume ratio of 1:1, adding pepsin to ensure that the activity value of the pepsin is 1800-2200U/mL, adjusting the pH value to 3 by using 1M hydrochloric acid, placing the solution in an oscillation incubator for 2 hours, wherein the temperature is 37 ℃, and the oscillation frequency is set to 80-150 r/min;
finally, mixing the mixed solution and simulated intestinal juice according to the volume ratio of 1:1, adding pancreatin to ensure that the activity value of the pancreatin is 80-120U/mL, adjusting the pH value to 7 by using 1M sodium hydroxide, placing the solution in an oscillation incubator for 2 hours at the temperature of 37 ℃ and the oscillation frequency of 80-150 r/min to prepare the artificial chyle, wherein the artificial chyle is preserved in a desk-top constant-temperature oscillator, the temperature is set at 37 ℃ and the oscillation frequency is set at 80-150 r/min.
In the step S3, the thickness of the intestinal mucus in the sample bottle is 0.5-1.5 cm, the thickness of the artificial chyle is 3-6 cm, and the intestinal mucus and the artificial chyle are added in sequence;
firstly, adding intestinal mucus into a sample bottle, then adding artificial chyle, slowly adding to avoid generating bubbles, and ensuring the liquid level to be level with a sample bracket without splashing the solution onto the bottle wall during adding.
A detection method of a chyle particle migration model in a mucus layer comprises the following steps:
s1, firstly, starting a computer and a multiple light scattering instrument, waiting for the self-checking of the instrument to be completed, and starting to run a test after the instrument is stabilized for 30 minutes;
s2, opening computer software to wait for the initialization of the instrument to be completed, then adjusting the detection temperature of the multiple light scattering instrument to 37 ℃, waiting for the instrument to rise to a specified temperature, pressing an open button to open the cover of the multiple light scattering instrument, putting the prepared sample bottle into the multiple light scattering instrument, pressing a close button to close the cover of the instrument, and taking care to avoid using hand pushing and not shaking the sample bottle vigorously in the sample placing process;
and S3, finally, a scanning program is newly established on a computer, the stabilizing time of the multiple light scattering instrument is set to be 2-10 min, the detection frequency is set to be 2-5 min for scanning once, the measuring time is set to be 8-12 h, the start button is clicked to start testing, and the migration of chylomicrons is analyzed through the fingerprint spectrum generated by the instrument.
(III) beneficial effects
The invention has the beneficial effects that:
1. the preparation and detection method of the chyle particle migration model in the mucus layer solves the problems of long experimental period, high operation difficulty, ethical concern and the like of conventional animals.
2. The preparation and detection method of the chyle particle migration model in the mucus layer has the advantages of short experimental period, relatively low technical requirements for operators, no ethical problem in the experimental process, and less cost.
3. According to the preparation and detection method of the chyle particle migration model in the mucus layer, not only can the color and shape changes of the sample be observed by naked eyes, but also the migration behaviors of the chyle particles can be comprehensively analyzed through the changes of the transmitted light and the back scattered light of the sample.
4. According to the preparation and detection method of the chyle particle migration model in the mucus layer, the model is simple to prepare, long-time continuous observation of a sample can be achieved, and the obtained result has certain representativeness and good repeatability.
Drawings
FIG. 1 is a graph of a transmitted light reference spectrum of the present invention;
FIG. 2 is a graph of a reference spectrum of the back-scattered light of the present invention.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
As shown in fig. 1-2, the present invention provides a technical solution: a preparation method of a chyle particle migration model in a mucus layer comprises the following steps:
s1, obtaining animal intestine mucus;
s2, preparing artificial chyle, and preserving under certain conditions;
s3, adding the intestinal mucus into a sample bottle, and then adding the artificial chyle.
In the step S1, the intestinal mucus is from the large intestine of a pig, the large intestine of the pig is fresh, preferably, the large intestine of the pig is taken immediately after slaughtering to extract the intestinal mucus, the intestinal mucus can be stored on ice cubes in the transportation process, the dirt on the surface of the large intestine of the pig is gently washed by PBS buffer solution before the intestinal mucus is extracted, but the large intestine of the pig is not required to be rubbed with force, the large intestine of the pig is sheared into small sections with the length of 15-20 cm after the washing is completed, the surface of the large intestine is rubbed with force to collect the mucus, a protease inhibitor is added into the intestinal mucus, and the intestinal mucus is stored at the temperature of minus 20 ℃;
in the step S2, firstly, fully dissolving a substance to be detected according to the required concentration, mixing the substance with simulated saliva according to the volume ratio of 1:1, adding salivary amylase to ensure that the enzyme activity value of the salivary amylase is 70-100U/mL, adjusting the pH value to be 7, and standing for 2-5 min at the temperature of 25 ℃;
mixing the mixed solution and simulated gastric juice according to the volume ratio of 1:1, adding pepsin to ensure that the activity value of the pepsin is 1800-2200U/mL, adjusting the pH value to 3 by using 1M hydrochloric acid, placing the solution in an oscillation incubator for 2 hours, wherein the temperature is 37 ℃, and the oscillation frequency is set to 80-150 r/min;
finally, mixing the mixed solution and simulated intestinal juice according to the volume ratio of 1:1, adding pancreatin to ensure that the activity value of the pancreatin is 80-120U/mL, adjusting the pH value to 7 by using 1M sodium hydroxide, placing the solution in an oscillation incubator for 2 hours at the temperature of 37 ℃ and the oscillation frequency of 80-150 r/min to prepare artificial chyle, and preserving the artificial chyle in a desk-top constant-temperature oscillator at the temperature of 37 ℃ and the oscillation frequency of 80-150 r/min;
in the step S3, the thickness of intestinal mucus in the sample bottle is 0.5-1.5 cm, the thickness of the artificial chyle is 3-6 cm, and the intestinal mucus and the artificial chyle are added in sequence;
firstly, adding intestinal mucus into a sample bottle, then adding artificial chyle, slowly adding to avoid generating bubbles, and ensuring the liquid level to be level with a sample bracket without splashing the solution onto the bottle wall during adding.
A detection method of a chyle particle migration model in a mucus layer comprises the following steps:
s1, firstly, starting a computer and a multiple light scattering instrument, waiting for the self-checking of the instrument to be completed, and starting to run a test after the instrument is stabilized for 30 minutes;
s2, opening computer software to wait for the initialization of the instrument to be completed, then adjusting the detection temperature of the multiple light scattering instrument to 37 ℃, waiting for the instrument to rise to a specified temperature, pressing an open button to open the cover of the multiple light scattering instrument, putting the prepared sample bottle into the multiple light scattering instrument, pressing a close button to close the cover of the instrument, and taking care to avoid using hand pushing and not shaking the sample bottle vigorously in the sample placing process;
and S3, finally, a scanning program is newly established on a computer, the stabilizing time of the multiple light scattering instrument is set to be 2-10 min, the detection frequency is set to be 2-5 min for scanning once, the measuring time is set to be 8-12 h, the start button is clicked to start testing, and the migration of chylomicrons is analyzed through the fingerprint spectrum generated by the instrument.
Example 1
A preparation method of a chyle particle migration model in a mucus layer comprises the following steps:
s1, obtaining animal intestinal mucus.
S2, preparing the artificial chyle, and preserving under certain conditions.
S3, adding the intestinal mucus into a sample bottle, and then adding the artificial chyle.
In the step S1, the large intestine of the pig is built into 15cm segments, the large intestine of the pig is washed by PBS buffer solution, then intestinal mucus is extracted, and the PBS buffer solution (pH 7.2-7.4): naCl 8g/L, KCl 0.2g/L, na 2 HPO 4 3.63g/L，KH 2 PO 4 The protease inhibitor Cocktail (100 x, DMSO stock) was added at a volume ratio to intestinal mucus of 1:100 at 0.24 g/L.
In the step S2, firstly, fully dissolving a substance to be detected according to the required concentration, mixing the substance with simulated saliva according to the volume ratio of 1:1, adding salivary amylase to ensure that the enzyme activity value of the salivary amylase is 70U/mL, adjusting the pH value to be 7, and standing for 2min at 25 ℃;
mixing the mixed solution and simulated gastric fluid according to the volume ratio of 1:1, adding pepsin to ensure that the activity value of the pepsin is 2000U/mL, adjusting the pH value to 3 by using 1M hydrochloric acid, placing the solution in an oscillation incubator for 2 hours at the temperature of 37 ℃ and setting the oscillation frequency to 100 r/min;
finally, mixing the mixed solution and simulated intestinal juice according to the volume ratio of 1:1, adding pancreatin to ensure that the activity value of the pancreatin is 100U/mL, adjusting the pH value to 7 by using 1M sodium hydroxide, placing the solution in an oscillation incubator for 2 hours at the temperature of 37 ℃ and setting the oscillation frequency to 100 r/min, and preparing the artificial chyle. The artificial chyle is stored in a desk type constant temperature oscillator, the temperature is set to 37 ℃, and the oscillation frequency is set to 100 r/min.
In the step S3, the thickness of intestinal mucus is 0.8cm, the thickness of the artificial chyle is 4cm, and the height of the added sample meets the requirement of detection on the height.
A detection method of a chyle particle migration model in a mucus layer comprises the following steps:
s1, firstly, turning on a computer and a multiple light scattering instrument;
s2, adjusting the detection temperature of the multiple light scattering instrument to 37 ℃, waiting for the instrument to rise to a specified temperature, and then placing the sample bottle into the multiple light scattering instrument, wherein the sample bottle is not required to be vigorously shaken in the sample placing process;
s3, setting the stabilizing time of the multiple light scattering instrument to be 5min, setting the detection frequency to be 3min for one scanning, and setting the measuring time to be 12h. And analyzing the migration of chylomicrons through the fingerprint spectrum generated by the instrument.
The experimental results are as follows with reference to fig. 1 and 2:
the migration of chyle particles was judged by the change in the height of intestinal mucus, and the data in the figure shows that the migration of chyle particles was completed after 8 hours, the height of intestinal mucus was increased by 1mm, and the precipitation rate of particles was calculated to be 0.125mm/h.
While the foregoing is directed to embodiments of the present invention, other and further details of the invention may be had by the present invention, it should be understood that the foregoing description is merely illustrative of the present invention and that no limitations are intended to the scope of the invention, except insofar as modifications, equivalents, improvements or modifications are within the spirit and principles of the invention.
Claims (1)
1. A preparation method of a chyle particle migration model in a mucus layer comprises the following steps:
s1, obtaining animal intestine mucus;
s2, preparing artificial chyle, and preserving under certain conditions;
s3, adding the intestinal mucus into a sample bottle, and then adding artificial chyle;
in the step S1, the intestinal mucus is from the large intestine of a pig, the large intestine of the pig is fresh, the large intestine of the pig is taken immediately after slaughter to extract the intestinal mucus, the intestinal mucus can be stored on ice cubes in the transportation process, the dirt on the surface of the large intestine of the pig is gently washed by PBS buffer solution before the intestinal mucus is extracted, but the large intestine of the pig is sheared into small sections with the length of 15-20 cm after the cleaning is finished, the surface of the large intestine is rubbed with force to collect the mucus, protease inhibitor is added into the intestinal mucus, and the intestinal mucus is stored at the temperature of minus 20 ℃;
in the step S2, firstly, fully dissolving a substance to be detected according to the required concentration, mixing the substance with simulated saliva according to the volume ratio of 1:1, adding salivary amylase to ensure that the enzyme activity value of the salivary amylase is 70-100U/mL, adjusting the pH value to be 7, and standing for 2-5 min at the temperature of 25 ℃;
mixing the mixed solution and simulated gastric juice according to the volume ratio of 1:1, adding pepsin to ensure that the activity value of the pepsin is 1800-2200U/mL, adjusting the pH value to 3 by using 1M hydrochloric acid, placing the solution in an oscillation incubator for 2 hours, wherein the temperature is 37 ℃, and the oscillation frequency is set to 80-150 r/min;
finally, mixing the mixed solution and simulated intestinal juice according to the volume ratio of 1:1, adding pancreatin to ensure that the activity value of the pancreatin is 80-120U/mL, adjusting the pH value to 7 by using 1M sodium hydroxide, placing the solution in an oscillation incubator for 2 hours at the temperature of 37 ℃ and the oscillation frequency of 80-150 r/min to prepare the artificial chyle, wherein the artificial chyle is preserved in a desk-top constant-temperature oscillator, the temperature is set at 37 ℃ and the oscillation frequency is set at 80-150 r/min;
in the step S3, the thickness of the intestinal mucus in the sample bottle is 0.5-1.5 cm, the thickness of the artificial chyle is 3-6 cm, and the intestinal mucus and the artificial chyle are added in sequence;
firstly, adding intestinal mucus into a sample bottle, then adding artificial chyle, slowly adding to avoid generating bubbles, and ensuring that the liquid level is level with a sample bracket without splashing the solution onto the bottle wall during adding;
when detecting a sample in a sample bottle, the detection method of the chyle particle migration model in the mucus layer comprises the following steps:
s1, firstly, starting a computer and a multiple light scattering instrument, waiting for the self-checking of the instrument to be completed, and starting to run a test after the instrument is stabilized for 30 minutes;
s2, opening computer software to wait for the initialization of the instrument to be completed, then adjusting the detection temperature of the multiple light scattering instrument to 37 ℃, waiting for the instrument to rise to a specified temperature, pressing an open button to open the cover of the multiple light scattering instrument, putting the prepared sample bottle into the multiple light scattering instrument, pressing a close button to close the cover of the instrument, and taking care to avoid using hand pushing and not shaking the sample bottle vigorously in the sample placing process;
and S3, finally, a scanning program is newly established on a computer, the stabilizing time of the multiple light scattering instrument is set to be 2-10 min, the detection frequency is set to be 2-5 min for scanning once, the measuring time is set to be 8-12 h, the start button is clicked to start testing, and the migration of chylomicrons is analyzed through the fingerprint spectrum generated by the instrument.
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| 失笑散合消糜汤加味治疗乳糜尿90例临床研究;殷学超;洪兵;陈丽娟;曹乃堂;王兆凯;;国医论坛;-;第-卷(第05期);21-22 * |
| 补脾固肾乳糜汤治疗乳糜尿;张国忠;;江西中医药;-;第-卷(第05期);35-36 * |
| 食物营养与机体健康的关系――肠胃始动论;阮征;邓泽元;伍国耀;印遇龙;;食品科学(第11期);273-282 * |
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