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Method for diagnosing and treating type II dentinogenesis by using DSPP gene and encoding product thereof

A dentin, incomplete technology, applied in the fields of bioengineering and medicine, can solve the problems of not fully revealing the exact cause, lack of effective methods for incomplete type II disease, and no one has revealed the correlation.

Inactive Publication Date: 2007-08-08
SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] However, until now, the exact cause of DI type II has not been fully revealed, nor has anyone revealed a direct correlation between DI type II and a certain protein
[0011] In addition, the art lacks an effective method for early and / or prenatal diagnosis of DI type II disease and an effective means for non-surgical treatment of DI type II

Method used

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  • Method for diagnosing and treating type II dentinogenesis by using DSPP gene and encoding product thereof
  • Method for diagnosing and treating type II dentinogenesis by using DSPP gene and encoding product thereof
  • Method for diagnosing and treating type II dentinogenesis by using DSPP gene and encoding product thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] There are 42 people in DGI-II families and 14 people in DGI-II families with deafness. All family members were carefully checked and diagnosed by experienced dentists, and detailed records were made. Deaf patients were carefully examined by ENT physicians and further confirmed by audiometry and brainstem evoked potentials. Take 5ml of peripheral blood from family members, anticoagulate with ACD blood cell preservation solution, and perform DNA extraction as follows:

[0079] Blood sample DNA preparation

[0080] Blood sample DNA was extracted using Qiagen kit, and the method was carried out according to the manufacturer's protocol. The specific steps are as follows:

[0081] a. Add proteinase K (20μl), anticoagulated blood sample (200μl) and buffer AL (200μl) into a 1.5ml centrifuge tube. Vortex for 15 seconds to mix.

[0082] b. After digesting at 56°C for 10 minutes, add 210 μl of 100% ethanol and centrifuge at low speed for 10 seconds.

[0083] c. Transfer the a...

Embodiment 2

[0091] 1. Genotyping:

[0092] Highly polymorphic STR markers in the 4q21 region (markers A-G are:

[0093] The primer sequences of D4s2691, D4s1534, GATA62A11, DSP, DMP1, SPP1, D4s451) were obtained from GenomeDatabase. PCR amplification was carried out in MJ-Research Inc. PTC-225 PCR instrument. The amplification program adopts the parameters of the touchdown Program in the operation manual of LI-COR Company. The PCR reaction system is 10μl, containing 20ng genomic DNA template, 2.0mM dNTP, 1.0pM positive strand primer with M13 tail and 1.0pM reverse strand primer, 1.0pM fluorescently labeled M13 primer, 1.5mM MgCl2, 10mM Tris-HCl, 1U Gold Taq enzyme (Perkin-Elmer Corp.). The reaction system begins with denaturation at 95°C for 8 minutes, followed by denaturation at 95°C for 45 seconds, annealing at 68°C for 2 minutes, decreasing by 2°C per cycle, and extending at 72°C for 1 minute. After 4 cycles, denaturation at 95°C for 45 seconds, annealing at 58°C for 1 minute , 72°...

Embodiment 3

[0103]Mutation Screening of Candidate Genes

[0104] Use Primer 5.0 (http: / / www / PrimerBiosoft.com) software to design primers covering the DSP gene exons and exons and intron junctions (the specific sequences are shown in Table 3), and use PCR-SSCP technology to The DSP gene was screened for mutations, and the PCR products were electrophoresed in 10% non-denaturing polyacrylamide gel and 9.3% non-denaturing polyacrylamide gel containing 4% glycerol, and then silver-stained according to conventional standard methods.

[0105] Primers are as follows:

[0106] Table 3 Primer sequences of DSPP coding region

[0107] Primer name

sequence

base number

DSPP-E1 Forward

5′-TGCAAAAGTCCATGACAGTG-3′

128

DSPP-E1 Reverse

5′-TCAGTTGGTTCTGAGTAAAAAGGA-3′

DSPP-E2 Forward

5′-AAGTAATTTTGTGCTGTTCCTTT-3′

149

DSPP-E2 Reverse

5′-AACAAAGTGAAGAGGTTTTCT-3′

DSPP-E3 Forward

5′-AAGAACCTTTTCAATTGCTAGT-3′

189

DSPP-...

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Abstract

The invention relates to a method for diagnosing dentinogenesis incomplete II type and / or dentinogenesis incomplete II type with deaf. The inventive method comprises that checking the DSPP gene, transfer sequence, and / or protein compared with normal ones; when the dissociation exists, the expression possibility of dentinogenesis incomplete II type and / or dentinogenesis incomplete II type with deaf of said person is higher than normal one. The invention also discloses a relative treatment method and drug.

Description

[0001] This application is a divisional application filed on September 5, 2000, and the application number is CN00125042.6. technical field [0002] The invention relates to the fields of bioengineering and medicine. More specifically, the present invention relates to a method for diagnosing and treating dentinogenesis imperfecta type II by using human DSPP gene and its coding product, as well as a pharmaceutical composition containing DSPP gene and / or protein. Background technique [0003] Dentin during tooth development and in mature teeth is produced by odontoblasts, which form dentine tubules during odontinogenesis. Odontocytic processes within the tubules make dentine a living tissue. In the initial stage of dentin formation, odontoblasts synthesize, secrete and reabsorb dentin matrix components. Protein synthesis occurs in the cell body, and exocytosis and endocytosis mainly occur in the cell process. The first to form is the unmineralized dentin matrix, which is mai...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/17A61P1/02A61P27/16C12Q1/68
Inventor 孔祥银肖尚喜赵国屏于川胡兰靛
Owner SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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