CN101275941A - Human gastric disease and animal model tissue chip - Google Patents
Human gastric disease and animal model tissue chip Download PDFInfo
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- CN101275941A CN101275941A CNA2007100384779A CN200710038477A CN101275941A CN 101275941 A CN101275941 A CN 101275941A CN A2007100384779 A CNA2007100384779 A CN A2007100384779A CN 200710038477 A CN200710038477 A CN 200710038477A CN 101275941 A CN101275941 A CN 101275941A
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Abstract
The present invention discloses a tissue microarray chip, and the tissue point whereon is composed of normal stomach tissue of human and animal, stomach tissue which is infected with helicobacter pylori and affection part tissue of stomach disease. Thereby the relationship between the whole process of generation and growth of the stomach disease and the infection of helicobacter pylori can be researched in the levels of molecule, cell and tissue. The chip also can screen out specific and significant gene and the protein expressed by the gene.
Description
One, technical field
The invention belongs to biological technical field, more specifically, relate on the sheet base and be fixed with organization chip.
Two, background technology
Along with histology, pathological development, people are to human body or other vegeto-animal understanding, and the organ level on anatomy develops to tissue, cellular level.But because complete organ is difficult to use microscopic examination, so need be cut into the very thin section observation of dyeing to organ,, histological techniques occur in order to address this problem.The principle of this method is by tissue that organ is cut into small pieces, after formalin fixed, utilize alcohol to slough moisture in this tissue, immerse dimethylbenzene again or benzole soln is sloughed alcohol, last piece of tissue immerses in the melted paraffin solution sloughs dimethylbenzene, put into mould and carry out embedding with paraffin solution, the cooling back forms paraffin mass.This paraffin mass utilizes microtome to cut into slices, and wax disk(-sc) is mounted on glass sheet, bakes sheet, dewaxing again, just can carry out observational study with microscope after dyeing, the mounting.At present, histological techniques is not only simple dyeing microscopic examination, various specific stains, immunohistochemistry technology, hybridization in situ technique, original position round pcr have also occurred.
In recent years along with genomics, protein science research and development, the physiology of people's research and disease progressively from molecular level to cellular level and organize the level development, therefore, how to utilize and organize research, be will one of deal with problems in the research at present.Adopt tradition to organize the Study on Technology method, because the tissue specimen requirement is big, the experimental work amount is big, and be difficult to adapt to research is wanted.In recent years, the tissue micro-array chip technology occurs, solved the problems referred to above.This technology fitly is arranged in dozens of, a hundreds of and even thousands of little tissue on a certain carrier (normally microslide) and is made into the micro histotomy, and its fundamental procedure (as shown in Figure 1) comprising: the relevant position, the organization chip that design and draft the making of the taxonomic series of organization chip, the pathological diagnosis of checking tissue or diseases related, tissue positioned, array wax stone, drill through tissue and transfer to blank paraffin mass again from the source wax stone are cut into slices.The characteristics of organization chip have: volume is little, information content is big, high flux; Reduce or avoided destruction the original structure wax stone; The accurate positioning of tissue samples reduces invalid tissue; Batch making and possible interpretation of result robotization; The processing of the case of separate sources, dyeing and analysis condition are identical.
Disease of stomach is one group of common, multiple disease of going.Though the cause of cancer of the stomach is at present also indeterminate, but it is roughly relevant: 1, chronic gastritis chronic gastritis with following several factors, particularly atrophic gastritis and cancer of the stomach is related closely, because atrophic gastritis often is associated with intestinal gland metaplasia and forms polyp, more increased the possibility that cancerates, therefore, usually atrophic gastritis is treated as precancerous lesion.2, Helicobacter pylori infection we usually the origin cause of formation of gastric ulcer was summed up as factors such as neuroendocrine in the past.Verified now, gastric ulcer is caused by helicobacter pylori.The gastric ulcer patient is carried out the regular treatment of system, to the very positive meaning that has of prevention cancer of the stomach.3. the polyp of stomach polyp of stomach belongs to the benign tumour of stomach, and the epithelial proliferation gonadoma in the polyp of stomach very easily cancerates into cancer, is considered to precancerous lesion.As find this disease, should active treatment to remove a hidden danger.4. the smoking and the smoking of drinking reach and drink for a long time, and particularly the great drinker generally has chronic gastritis, and becomes between chronic gastritis may take place just, even cancerates into cancer of the stomach, also is an important step of prevention cancer of the stomach thus away from tobacco and wine.5. dietary factor 6. inherent cause cancer of the stomach have family's tendentiousness clearly.Early carcinoma of stomach can be divided into following various (nineteen ninety WHO classification): 1. gland cancer comprise papillary adenocarcinoma, tubular adenocarcinoma, with myxoadenocarcinoma (grade of malignancy is higher than the above two), be divided into high differentiation, middle differentiation and 3 kinds of low differentiation again according to differentiation degree again; 2. undifferentiated carcinoma (grade of malignancy is the highest); 3. carcinoma muco-cellulare (being signet ring cell cancer); 4. specific type cancer: comprise adenosquamous carcinoma, squama cancer, carcinoid, suede cancer, carcinosarcoma and neuroendocrine carcinoma etc.Also have some rare cancers: liver cancer sample cancer of the stomach, parietal cell cancer, cephaloma etc.There is the variation of an active development in various diseases: carcinoma in situ (primary carcinoma is limited to mucous layer and does not involve proper mucous membrane); Tumour is invaded and submucosa; Tumour is invaded and the flesh layer; Tumour is invaded and placenta percreta; Invade and adjacent organs; Gastroscopic biopsy turns out to be cancer, but cancer is not found in the histological examination of excision sample; Tumor invasive depth can't be judged; No regional lymph node metastasis; Lymphatic metastasis is confined to borderline tumor 3cm with interior stomach week lymph node; Lymphatic metastasis exceeds beyond the borderline tumor 3cm, comprises left gastric artery, arteria hepatica communis, spleen and abdominal cavity peripheral lymph node; The lymphatic metastasis situation can't be determined; No DISTANT METASTASES IN etc.
The subordinate's of the World Health Organization (WHO) in 1994 IARC is classified helicobacter pylori as I class (affirming) procarcinogen that causes cancer of the stomach.The generation development how helicobacter pylori causes tumour is the difficult point and the focus of research at present.Seek a kind of effectively reliably research tool, thereby from molecule, cell, organize that level is observed the generation of these diseases, the whole process of development finds significant gene and expressed protein, is present urgent problem.
Three, summary of the invention
The purpose of this invention is to provide a kind of efficient, easy and gastric tissue mixings micro-array chip of people and the animal that infects helicobacter pylori cancer of the stomach tumor susceptibility gene cheaply, thus can be at molecule, cell, organize and study cancer of the stomach disease and the generation of helicobacter pylori in human body and animal body, the whole process of development on the level.By this chip also can filter out specificity, significant gene and and expressed protein as the potential use of diagnosis and treatment.
This gastric tissue micro-array chip comprises being connected with the specific tissue point on sheet base, the sheet base.The sheet base can be glass, biological membrane, silicon, poly-difluoride membranes or nitrocellulose membrane, and the sheet base can be to handle through anti-flake.The required different size diameter of interlacing point can be 0.6mm, 1.0mm, 1.5mm, 2.0mm, 2.5mm, 3.0mm, 3.5mm, 4.0mm, 4.5mm, 5.0mm, 5.5mm, 6.0mm etc. on the sheet base.Density is 500 point/square centimeter～1 point/square centimeters, can have 1000 point～10 not wait on every sheet base.Can the rectangular array mode, circular, fashion or other shapes arrange.
Described interlacing point is selected from the human stomach tissue and the animal model gastric tissue of following normal or disease pathology at least: the human stomach tissue (comprises at the bottom of the stomach, body of stomach, lesser curvature, greater curvature, pylorus, orifice of the stomach) normal structure, chronic superficial gastritis, the simple form atrophic gastritis, gastric ulcer, intestines type atrophic gastritis completeness colonization is given birth to, intestines type atrophic gastritis imperfection colonization is given birth to, intestines type atrophic gastritis completeness enterization is given birth to, and intestines type atrophic gastritis imperfection enterization is given birth to, the stomach atypical hyperplasia, stomach tube shape adenoma, villous adenoma of stomach, stomach tube shape villous adenoma, ZoHinger-Ehison syndrome, stomach vascular knurl, leiomyoma of stomach, leiomyosarcoma of stomach, nervus gastrica sheath knurl, the stomach malignant schwannoma, the stomach papillary adenocarcinoma, stomach tube shape gland cancer, gastric mucus gland cancer, the stomach signet ring cell cancer, the stomach adenosquamous carcinoma, stomach squama cancer, stomach small cell carcinoma, the stomach undifferentiated carcinoma, malignant lymphoma of stomach etc.; The gastric tissue of the normal and pathology of animal model: at first set up helicobacter pylori infections animal stomach model.Zoologize the feed that contains helicobacter pylorus strain as Mongolian gerbil jird feeding and infect by giving, in 1 week, the different time periods such as 2 weeks or 3 weeks slaughter animal and obtain gastric tissue, zoologize and inducing the whole process that causes gastric tissue generation pathology, finally cause cancer of the stomach to take place through a series of environmental factor.The tissue of choosing comprises: stomach (at the bottom of comprising stomach, body of stomach, lesser curvature, greater curvature, pylorus, orifice of the stomach) normal structure, inflammation tissue, precancerous lesion tissue and cancerous issue.
Specifically set forth content of the present invention below in conjunction with embodiment
One, people and animal incidence gastric cancer mechanism and development overall process organization chip
The tissue type information of each point is shown in form one in people and animal incidence gastric cancer mechanism and the development overall process organization chip array, arrange organization chip design sketch that the back forms as shown in Figure 2: 1 for handle through anti-flake at slide, 2 are carbon core gauge point (mark array reads order), 3 tissue samples for arrangement.The diameter of point is 1.0mm, and the spacing between the point is 0.4mm.10 points of every row, totally 12 row.Types of organization below in conjunction with arrangement point in the form one enumeration array: have a carbon core gauge point (Maker) to be used for positioning in the chip upper left corner to the orientation of array.A1, A2, for infecting the male mongolian gerbil gastric tissue in ATCC 3 weeks of bacterial strain; A3, A4, for infecting the female mongolian gerbil gastric tissue in ATCC 3 weeks of bacterial strain; A5, A6, A7, A8, for male mongolian gerbil gastric tissue A9, the A10 that infects ATCC 8 weeks of bacterial strain be not infectious bacteria and raise the mongolian gerbil gastric tissue in 8 weeks of control group.B1, B2, B3, B4 are for infecting the male mongolian gerbil gastric tissue in ATCC 3 weeks of bacterial strain; A3, A4, for infecting the female mongolian gerbil gastric tissue in ATCC 3 weeks of bacterial strain; A5, A6, A7, A8, for the male mongolian gerbil gastric tissue C5, the C6 that infect ATCC 8 weeks of bacterial strain, be control group infectious bacteria and raise the mongolian gerbil gastric tissue in 20 weeks not; C7, C8, C9, C10 are for infecting the male mongolian gerbil gastric tissue in HP161 8 weeks of bacterial strain.D1, D2, for infecting the male mongolian gerbil gastric tissue in HP161 8 weeks of bacterial strain; D3, D4, be control group infectious bacteria and raise the mongolian gerbil gastric tissue in 8 weeks not; D5, D6, D7, D8, for infecting the female mongolian gerbil gastric tissue in HP161 8 weeks of bacterial strain; D9, D10 are not infectious bacteria and raise the mongolian gerbil gastric tissue in 8 weeks of control group.E1, E2, for infecting the male mongolian gerbil gastric tissue in HP161 20 weeks of bacterial strain; E3, E4, be control group infectious bacteria and raise the mongolian gerbil gastric tissue in 20 weeks not; E5, E6, E7, E8, for infecting the female mongolian gerbil gastric tissue in HP161 20 weeks of bacterial strain; E9, E10 are the infectious bacteria and raise the mongolian gerbil gastric tissue in 20 weeks not of control group not.F1, F2, F3, F4 are respectively stomach (orifice of the stomach), stomach (big curved), stomach (little curved), stomach (hole) normal structure; F5, F6, F7 are respectively chronic superficial gastritis evolution mild or moderate, moderate, the pathological tissues of stressed three phases; F8, F9, F10 are respectively simple form atrophic gastritis evolution mild or moderate, moderate, the pathological tissues of severe three phases; G1 is gastric ulcer pathology tissue; G2, G3, G4, G5 are respectively the different causes of disease of intestines type atrophic gastritis: the completeness colonization is given birth to, and the imperfection colonization is given birth to, and the completeness enterization is given birth to, and the imperfection enterization is given birth to four kinds of pathological tissues; It is slight that G6, G7, G8 are respectively three different stages of development of stomach atypical hyperplasia, moderate, the pathological tissues of severe; G9, G10, H1, H2, H3, H4, H5, H6, H7 are respectively stomach tube shape adenoma, villous adenoma of stomach, stomach tube shape villous adenoma, ZoHinger-Ehison syndrome, stomach vascular knurl, leiomyoma of stomach, leiomyosarcoma of stomach, nervus gastrica sheath knurl, stomach malignant schwannoma cancerous issue; H8, H9, H10, I1 are respectively tumor tissues and the other tissue of tumour that two examples suffer from stomach papillary adenocarcinoma patient; I2, I3, I4, I5 are respectively tumor tissues and the other tissue of tumour that two examples suffer from stomach tube shape gland cancer patient; I6, I7, I8, I9 are respectively tumor tissues and the other tissue of tumour that two examples suffer from gastric mucus gland cancer patient; I10, J1, J2, J3 are respectively tumor tissues and the other tissue of tumour that two examples suffer from stomach signet ring cell cancer patient; J4, J5, J6, J7 are respectively tumor tissues and the other tissue of tumour that two examples suffer from stomach adenosquamous carcinoma patient; J8, J9, J10, K1 are respectively tumor tissues and the other tissue of tumour that two examples suffer from stomach squama cancer patient; K2, K3, K4, K5 are respectively tumor tissues and the other tissue of tumour that two examples suffer from stomach small cell carcinoma patient; K6, K7, K8, K9 are respectively tumor tissues and the other tissue of tumour that two examples suffer from stomach undifferentiated carcinoma patient; K10, L1, L2, L3 are respectively tumor tissues and the other tissue of tumour that two examples suffer from the malignant lymphoma of stomach patient; L4 is that a carbon core gauge point (Marker) is used for the end position that mark array is arranged.The chip normal and the canceration control tissue of this Zhang Hanyou reflection people and animal cancer of the stomach different onset mechanism and development overall process is used for SABC, in situ hybridization, different tests researchs such as immunofluorescence.
Description of drawings:
Fig. 1 makes basic procedure for organization chip.
Fig. 2 be the organization chip synoptic diagram wherein: 1 for handle through anti-flake at slide; 2 are carbon core gauge point (mark array reads order); 3 tissue samples for arrangement.
Should be understood that the exercise question of this specification and the embodiment that this specification is addressed only are used for explanation The present invention and being not used in limits scope of the present invention.
Claims (10)
1. tissue micro-array chip, comprise sheet base and interlacing point, interlacing point is arranged and is invested on the sheet base with dot matrix way, it is characterized in that described interlacing point is made of the mongolian gerbil stomach tissue of normal human subject normal gastric mucosa, human gastric disease pathology portion tissue, the normal stomach tissue of mongolian gerbil, infection helicobacter pylorus.
2. a kind of tissue micro-array chip according to claim 1 is characterized in that described base can be glass, film, biological membrane, silicon, poly-difluoride membranes or nitrocellulose membrane.
3. a kind of tissue micro-array chip according to claim 2 is characterized in that described base can be to handle through anti-flake.
4. a kind of tissue micro-array chip according to claim 1 is characterized in that, described interlacing point diameter is 0.6 millimeter, 1.0 millimeters, and 1.5 millimeters, 2.0 millimeter, 2.5 millimeters, 3.0 millimeters, 3.5 millimeters, 4.0 millimeters, 4.5 millimeter, 5.0 millimeters, 5.5 millimeters, 6.0 millimeters.
5. a kind of tissue micro-array chip according to claim 1 is characterized in that described dot matrix way can arrange for rectangular array mode or garden shape mode, and density is 500 to 1 point/square centimeters, and quantity is 1000 o'clock to 10 o'clock.
6. a kind of tissue micro-array chip according to claim 1 is characterized in that the human normal gastric mucosa of being addressed comprises human stomach orifice of the stomach, lesser curvature, greater curvature, stomach hole portion tissue at least.
7. a kind of tissue micro-array chip according to claim 1, it is characterized in that the gastric disease of being addressed comprises chronic superficial gastritis at least, simple gastritis, atrophic gastritis, gastric ulcer, the stomach atypical hyperplasia, stomach tube shape adenoma, villous adenoma of stomach, stomach tube shape villous adenoma, ZoHinger-Ehison syndrome, stomach vascular knurl, leiomyoma of stomach, leiomyosarcoma of stomach, nervus gastrica sheath knurl, the stomach malignant schwannoma, the stomach papillary adenocarcinoma, stomach tube shape gland cancer, gastric mucus gland cancer, the stomach signet ring cell cancer, the stomach adenosquamous carcinoma, stomach squama cancer, the stomach small cell carcinoma, stomach undifferentiated carcinoma and malignant lymphoma of stomach.
8. a kind of tissue micro-array chip according to claim 1 is characterized in that the stomach tissue of the infection helicobacter pylorus mongolian gerbil addressed comprises mongolian gerbil stomach tissue that infects the ATCC bacterial strain and the mongolian gerbil stomach tissue that infects the HP161 bacterial strain.
9. a kind of tissue micro-array chip according to claim 8 is characterized in that the infection ATCC bacterial strain mongolian gerbil stomach tissue of being addressed comprises the male mongolian gerbil gastric tissue that infects ATCC 3 weeks of bacterial strain, the female mongolian gerbil gastric tissue that infects ATCC 3 weeks of bacterial strain, the male mongolian gerbil gastric tissue in infection ATCC 8 weeks of bacterial strain.
10. a kind of tissue micro-array chip according to claim 8, it is characterized in that described infection HP161 bacterial strain mongolian gerbil stomach tissue comprise male mongolian gerbil gastric tissue, the male mongolian gerbil gastric tissue that infects HP161 8 weeks of bacterial strain, the female mongolian gerbil gastric tissue that infects HP161 8 weeks of bacterial strain that infects HP161 8 weeks of bacterial strain, the male mongolian gerbil gastric tissue that infects HP161 20 weeks of bacterial strain,, the female mongolian gerbil gastric tissue in infection HP161 20 weeks of bacterial strain.
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| CNA2007100384779A CN101275941A (en) | 2007-03-26 | 2007-03-26 | Human gastric disease and animal model tissue chip |
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| CNA2007100384779A CN101275941A (en) | 2007-03-26 | 2007-03-26 | Human gastric disease and animal model tissue chip |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2723895A2 (en) * | 2011-06-23 | 2014-04-30 | Children's Hospital Medical Center | Molecular diagnostic panel of eosinophilic gastrointestinal disorders |
| CN108037278A (en) * | 2017-12-13 | 2018-05-15 | 广州吉奥生物科技有限责任公司 | The preparation method of immunohistochemistry detection section |
| CN112614128A (en) * | 2020-12-31 | 2021-04-06 | 山东大学齐鲁医院 | System and method for assisting biopsy under endoscope based on machine learning |
| US11564905B2 (en) | 2016-01-13 | 2023-01-31 | Children's Hospital Medical Center | Compositions and methods for treating allergic inflammatory conditions |
| US11859250B1 (en) | 2018-02-23 | 2024-01-02 | Children's Hospital Medical Center | Methods for treating eosinophilic esophagitis |
-
2007
- 2007-03-26 CN CNA2007100384779A patent/CN101275941A/en active Pending
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2723895A2 (en) * | 2011-06-23 | 2014-04-30 | Children's Hospital Medical Center | Molecular diagnostic panel of eosinophilic gastrointestinal disorders |
| EP2723895A4 (en) * | 2011-06-23 | 2015-02-11 | Childrens Hosp Medical Center | Molecular diagnostic panel of eosinophilic gastrointestinal disorders |
| US9345763B2 (en) | 2011-06-23 | 2016-05-24 | Children's Hospital Medical Center | Methods of treating allergic inflammatory conditions by administering an anti-cadherin-like 26-based therapeutic |
| US9803244B2 (en) | 2011-06-23 | 2017-10-31 | Children's Hospital Medical Center | Methods of determining eosinophilic gastritis status based on marker or gene expression |
| US11564905B2 (en) | 2016-01-13 | 2023-01-31 | Children's Hospital Medical Center | Compositions and methods for treating allergic inflammatory conditions |
| CN108037278A (en) * | 2017-12-13 | 2018-05-15 | 广州吉奥生物科技有限责任公司 | The preparation method of immunohistochemistry detection section |
| CN108037278B (en) * | 2017-12-13 | 2018-09-25 | 广州吉奥生物科技有限责任公司 | The preparation method of immunohistochemistry detection slice |
| US11859250B1 (en) | 2018-02-23 | 2024-01-02 | Children's Hospital Medical Center | Methods for treating eosinophilic esophagitis |
| CN112614128A (en) * | 2020-12-31 | 2021-04-06 | 山东大学齐鲁医院 | System and method for assisting biopsy under endoscope based on machine learning |
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Open date: 20081001 |