Thirty patients of thalassemia major (8–18 years old) receiving packed cells transfusion, to maintain hemoglobin concentration between 9.0 to 10.5 g/dl, were selected for the study. Results were compared with a group of fifteen normal healthy controls (group I). Patients were subdivided into two groups, whereas every alternate patient attending the Thalassemia Clinic and receiving packed cell transfusion therapy was included in group II, the remaining 15 patients were included in group III. The patients in group III, in addition to packed cell transfusion, were supplemented with calcium (1.5 g/day, for 3 months) and vitamin D3 (60,000 IU/day, for 10 days). These patients were again re-evaluated after 3 months of the supplementation of the two micronutrients.
All the subjects were assessed for routine anthropometric and laboratory investigations that were found to be within normal limits. Mean age of the children in different groups was also comparable and was 11.87 ± 2.13 years for group I, 12.01 ± 2.82 years for group II, and 11.61 ± 2.89 years for group III, respectively. The three groups were also sex matched.
All the patients were on regular PCV infusion since the age of 1–2 years, after the diagnosis of thalassemia was confirmed. Inclusion criteria were—(a) age 8–18 years, (b) diagnosis of thalassemia major, and (c) those recieving regular PCV infusion and chelation therapy. Exclusion criteria were—(a) poor compliance, (b) very sick patients, (c) those that cannot be followed, (d) having some renal disease, (e) malabsorption syndrome, or (f) on long term anticonvulsant therapy. Informed consent of the parent was obtained. Approval of IRB and ethical committee were also taken.
Five ml venous blood was collected in a plain vial, asceptically. Serum was separated and analyzed immediately (except for PTH), using Konelab 30i Clinical Chemistry Analyzer. For PTH, serum was frozen and used within a week. Concentrations of serum and 24 h urinary calcium were estimated using O-cresolpthalein complexone [6
], serum phosphorus according to Fiske and Subbarow [7
], alkaline phosphatase according to King and Armstrong [8
], and albumin by dye-binding method using bromocresol green [9
]. Albumin-corrected calcium was calculated (to obtain a reasonable estimate of the free calcium) according to Pyane [10
], who suggested that albumin-corrected calcium gives an estimate of the free calcium level when albumin levels are within the normal range. Serum intact PTH was estimated by two-site labeled antibody radioimmuno assay [11
]. Results were statistically analyzed by Student’s t