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Pattern of vitamin d receptor polymorphism
- 4. Age-related loss of trabecular bone
- 6. Functions of the Vitamin D Endocrine System
- 8. Schematic diagram of the human vitamin D receptor gene
- 12. DNA Amplification with the Polymerase Chain Reaction (PCR) Make copies ( extend primers ) Starting DNA Template 5’ 5’ 3’ 3’ 5’ 5’ 3’ 3’ Add primers ( anneal ) 5’ 3’ 3’ 5’ Forward primer Reverse primer Separate strands ( denature ) 5’ 5’ 3’ 3’
- 13. In 35 cycles at 100% efficiency, 1.07 billion copies of targeted DNA region are created PCR Copies DNA through Multiple Thermal Cycles Original DNA target region Thermal cycle Thermal cycle Thermal cycle
- 14. Mutation detection by restriction analysis 7 6 5 4 3 2 1 Ladder 69 bp-- 196 bp-- 265 bp--
- 15. 7 6 5 4 3 2 1 Ladder 69 bp-- 196 bp-- 265 bp--
- 16. 8 7 6 5 4 3 2 1 Ladder 200 bp 650 bp 850 bp
- 17. RESULTS
- 18. Body mass index of osteoporotic and controls
- 19. Distribution of different genotypes of FOKI polymorphism & alleles frequency in patients & controls: FF genotype Ff genotype ff genotype Allele (%) F f Patients (n=50) N (%) 34 (68) 9 (18) 7 (14) 72.9 27.1 Controls (n=20) N (%) 20 (100) 0 (0) 0 (0) 100 0
- 20. Distribution of different genotypes of BSMI polymorphism & alleles frequency in patients & controls: BB genotype Bb genotype bb genotype Allele (%) B b Patients (n=50) N (%) 27 (54) 15 (30) 8 (16) 64.6 35.4 Controls (n=20) N (%) 1 (5) 2 (10) 17 (85) 13.6 86.4
- 21. Comparison of the studied parameters in FOKI 0.07 0.68±.088 .74 ± .140 BMD lumbar (g/cm 2 ) 0.000 0.68±.040 0.77 ± .129 BMD femoral (g/cm 2 ) 0.04 -2.11 ±0 .91 -1.54 ± 1.14 T score lumbar 0.3 -1.21 ± 1.14 -.94 ± .983 T scorefemoral P-value Ff+ff (N=16) Mean ± SD FF (N=54) Mean ± SD Variables
- 22. BMD lumbar of osteoporotic patients
- 23. Comparison of the studied parameters in BSMI 0.000 0.67±0.10 0.84 ± 0.11 BMD lumbar (g/cm 2 ) 0.000 0.70 ±0 .07 0.85 ±0 .13 BMD femoral (g/cm 2 ) 0.001 -2.00 ± 1.02 -1.08 ± 1.05 T score lumbar 0.10 -1.14 ± 1.06 -0.75 ± 0 .92 T scorefemoral P-value Bb+bb (N=45) Mean ± SD bb (N=25) Mean ± SD Variables
- 24. BMD lumbar of osteoporotic patients
- 25. Combined genotypes of BSMI and FOKI of osteoporotic patients
- 26. Combined genotype of BSMI and FOKI of controls
- 27. O.R ratio of FOK1genotypes of patients and controls: (32) (0) (%) 16 0 Count Ff+ff (68) (100) (%) 1.588(1.29-1.95) 0.004 34 20 Count FF cases control FOK1 OR (95% CI) P value GROUP
- 28. O.R ratio of BSM1 genotypes of patients and controls: (16) (85) (%) 8 17 Count bb (84) (15) (% ) 29.75(7.04-125.8) 0.000 42 3 Count BB+Bb cases control BSM1
- 31. Thank you
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- extraction of DNA using salting out technique followed by amplification of extracted DNA at exon 2 for FOKI & intron 8 for BSMI and amplified DNA was then cut by restriction enzyme FOKI for detection of FOKI polymorphism and by enzyme BSMI for detection of BSMI polymorphism and then electrophoresis of the DNA was done on agarose gel.