1.Measurement Of β-and γ-mRNA Levels In Thalassemia By Fluorescence Quantitative Reverse Transcription Polymerase Chain Reaction 2.Umbilical Cord Blood Transplantation For A Patient With β-thalassemia Major

Objective 1. To study the application of fluorescence quantitative reverse transcription polymerase chain reaction (FQ-RT- PCR) in the measurement of hemoglobin mRNA levels. 2. To investigate the P - and Y -mRNA levels in patients with thalassemia and umbilical cord blood. Methods We measured the P - and y -mRNA levels from red cells of 38 patients with thalassemia, 25 normal controls and 23 newborn umbilical cord blood by the FQ- RT-PCR. Results All of the 17 patients with P -thalassemia major had an increased y P + v) mRNA ratio .The average i P + y) mRNA ratio in the P -thalassemia majors (70.83~22. 17%) was significantly higher than that of normal controls (1.5 5~4.l4%) (p<0.O0l) and the P -thalassemia minors (0.9% 1 .01%) (j<0.00 1). There was no significant difference between the P -thalassemia minors and the normal controls (j,>0.05). The mean of y I( P + y) mRNA ratio in the newborn umbilical cord blood (44.08d:22.4 1%) was significantly higher than that of normal controls (p(O.0O 1). The mean of J-LbF ratio in the newborn umbilical cord blood (58.1 1~7. 19%) was significantly higher than that of normal controls (O.63~0.32%) (P<0.00l). FQ-RT-PCR was in-tube monitoring and eliminating post-PCR processing, which removed the 9. chances for carryover contamination and the potential sources of errors. 23 A real-time detection ensures accurate quantity. Conclusion 1. FQ-RT-PCR is a new method suitable for the measurement of hemoglobin mRNA levels, which has a high-throughput, increased reproducibility and precision qualities. 2. The average ~?/( P + mRNA ratio in the P -thalassemia majors was significantly higher than that of normal controls and P -thalassemia minors .An increased y /( ~ + Y) mRNA ratio is one of the features of P -thalassemia major. There was no significant difference between the P -thalassemia minors and the normal controls. Measurement of the P - and y -niRNA levels is helpful for the diagnosis and therapeutic effects observation in thalassemia. 3. The mean of i?/( P + y) mRNA ratio and HbF rate in the cord blood were significantly higher than those of normal controls. Analyzing the hemoglobin mRNA levels in the umbilical cord blood can help to understand the control of hemoglobin gene expression.