J Endocrinol Metab

Journal of Endocrinology and Metabolism, ISSN 1923-2861 print, 1923-287X online, Open Access

Article copyright, the authors; Journal compilation copyright, J Endocrinol Metab and Elmer Press Inc

Journal website http://www.jofem.org

Original Article

Volume 1, Number 4, October 2011, pages 174-181

Compound Heterozygous Mutations in Vitamin D Receptor Gene in Two Sisters With Hereditary Vitamin D Resistant Rickets Type II

Figures

Figure 1. X-ray A) wrist and B) lower limb of the first patient showed radiological signs of active rickets: diffuse osteoporosis, widening of the zone of provisional calcification at the metaphysis, metaphyseal fraying, flaying and cupping of proximal and distal tibia, distal femur, radius and ulna, irregular ill defined outline of the epiphysis, genu valgum deformity, bilateral anterior bowing of the lower part of the tibia and fibula, and delayed bone age. C) Improvement of signs of active rickets and of bone age after treatment.

Figure 2. Electropherogram of exon 2 of the VDR gene: A is the first patient showing A to C substitution at position 22, B is the father with normal base sequence.

Figure 3. Electropherogram of exon 3 of the VDR gene: A is the first patient with a substitution and a deletion mutations, at nucleotide position 150 of the VDR gene, B is the father with the substitution mutation.

Tables

**Table 1.** Biochemical and hormonal profiles of the 2 patients
	Patient 1			Patient 2			Reference value
	Jan/08	Feb/09	Jul/11	Jan/08	Feb/09	Jul/11	Reference value
*urinary phosphate.
Ca (mg/dl)	9.7	9.7	9.5	9.4	8.9	10.4	8.8 - 10.8
P (mg/dl)	2.9	2.8	2.9	2.1	3.9	5.6	3.7 - 5.6
ALP (U/L)	1035	960	422	1573	767	392	145 - 420
PTH (pg/ml)	118	162	60	620	315	38.4	12 - 95
U. PO₄ (g/24h/l)*	-	0.4	-	-	0.2	-	0.4 - 1.3
1,25(OH)₂D₃ (pg/ml)	74	106	97	71	75	146	27 - 71
25(OH)D (ng/ml)	-	14	18	-	7	14	20 - 200

**Table 2.** Primers and PCR conditions used for amplifications
	Primers	PCR conditions
Exon 2	2 a sense, 5'-AGCTGGCCCTGGCACTGACTCTGCTCT-3' 2 b antisense, 5'- ATGGAAACACCTTGCTTCTTCTCCCTC-3'	1) Denaturation at 94 °C for 5 min. 2) 35 cycles of denaturation at 94 °C for 1 min., annealing at 60 °C for 1 min., and extension at 72 °C for 1 min.
Exon 3	3a sense, 5'- AGGGTGAAGGAGCCGGAAGTTCAGTGAC-3' 3b antisense, 5'-CTTTCCCTGACTCCACTTCA GGCCCAA-3'	3) final extension at 72 °C for 10 min.
Exons 7& 8	7a sense 5'-GCGAATTCCGTTACTGGTAACCTGACCTCTTC-3' 8b antisense 5'-TGGAATTCATACACCCCGCTCCCCAGTCCCTGAG-3'	As above but with annealing at 56 °C.