We aimed to establish a method for quantitative analysis of mixed haematopoietic chimerism based on microchip electrophoresis of selected molecular markers following PCR amplification for accurate monitoring of graft status post-transplantation. A 12-year-old girl with relapsed acute lymphoblastic leukaemia who underwent allogeneic bone marrow transplantation had qualitative chimerism analysis using short tandem repeat markers at three time points following the procedure. Her archived DNA samples were then used to test the ability to correlate her clinical course with changes in the quantity of donor chimerism at the different time points. Quantitative chimerism analysis was performed on the Agilent 2100 bioanalyser and donor-recipient ratios were calculated from generated electropherograms. Complete donor chimerism (98%) was demonstrated three weeks post- transplantation. Decreasing amount of donor chimerism to 24% was shown after three months and this concurred with clinical relapse. Following a second transplant, full donor chimerism was reestablished where donor chimerism rose to 100%. High resolution microchip electrophoresis could be useful in predicting the occurrence of increasing recipient chimerism which may herald impending relapse in patients while the disease burden is still low. This investigational approach may provide useful information for clinicians to select appropriate intervention strategies to ensure successful transplantation.

Midazolam is one of the most commonly used drugs for sedation in Emergency Department (ED). This was a retrospective study conducted on 380 patients from December 2012 to May 2014 in ED of Universiti Kebangsaan Malaysia Medical Centre (UKMMC). The objective was to elicit the frequency of side effects and correlation to various factors i.e. socio-demography, co-morbidities, age groups and underlying illnesses. Out of 380 patients, 35 patients experienced side effects (20 patients with midazolam alone, 15 patients with combination of drugs). The average age was 42 years and the average dose of midazolam was 3.5mg. The most common other drug combined was fentanyl. The overall complication rate for midazolam was 5.3%. The most common side effect recorded was excessive somnolence (1.6%). Other side effects included local skin reactions (1.1%), vomiting (0.8%), headache (0.8%) and hypotension (0.5%). There was no significant association between the socio-demographic factors and drugs combination with the side effects of midazolam on patients. It was concluded that midazolam was a safe drug due to absence of any life-threatening side effects. There are possibilities that most side effects recorded could be caused by other comfounding factors e.g. underlying injuries or disease and combination with other drugs.
Midazolam

In childhood acute lymphoblastic leukaemia (ALL), cytogenetics play an important role in diagnosis, allocation of treatment and prognosis. Conventional cytogenetic analysis, involving mainly karyotyping in our experience, has not been successful in a large proportion of cases due to inadequate metaphase spreads and poor chromosome morphology. Our aim is to develop a highly sensitive and specific method to screen simultaneously for the four most frequent fusion transcripts resulting from specific chromosomal translocations, namely, both the CML- and ALLtype BCR-ABL transcripts of t(9;22), E2A-PBX1 transcript of t(1;19), the MLL-AF4 transcript of t(4;11) and TEL-AML1 (also termed ETV6-CBFA2) of the cryptic t(12;21). A multiplex reverse transcription polymerase chain reaction protocol (RT-PCR) was developed and tested out on archival bone marrow samples and leukaemia cell lines. In all samples with a known translocation detected by cytogenetic techniques, the same translocation was identified by the multiplex-PCR assay. Multiplex RT-PCR assay is an effective, sensitive, accurate and cost-effective diagnostic tool which can improve our ability to accurately and rapidly risk-stratify patients with childhood ALL.
Child ; Humans ; Oncogene Proteins, Fusion ; genetics ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Translocation, Genetic

INTRODUCTIONThalassaemia is one of the major public health problems in Malaysia. Regular monthly blood transfusion remains the main treatment for severe thalassaemia patients. One of the complications of blood transfusion is the formation by the recipients of alloantibodies and autoantibodies against red blood cell (RBC) antigen. The purpose of this study was to determine the prevalence of RBC autoantibodies among multiple-transfused thalassaemic patients in our institution and factors that contribute to its development.

METHODSA prospective study was conducted in Haematology Laboratory, Hospital Universiti Sains Malaysia between January 2004 and December 2004. A total of 63 thalassaemia patients, who received regular blood transfusion were included in this study. Clinical and serological data were collected and analysed prospectively. Blood samples were subjected to standard blood bank procedures for screening of antibodies and their subsequent identification using reagent of Diamed-ID Gel microtyping system.

RESULTSThere were 49 (77.8 percent) patients with Hb E/beta-thalassaemia, ten (15.9 percent) beta-thalassaemia major, three (4.7 percent) Hb H Constant Spring and one (1.6 percent) Hb H disease. Only one (1.6 percent) patient had autoantibodies. There were no statistical associations found between the formation of autoantibodies with age at the start of transfusion, number of packed cell transfused and splenectomy.

CONCLUSIONOur data showed a low autoimmunisation rate in multiple-transfused thalassaemia patients in our hospital.

Adolescent ; Adult ; Autoantibodies ; blood ; Blood Group Antigens ; immunology ; Child ; Child, Preschool ; Coombs Test ; Erythrocytes ; immunology ; Female ; Hospitals, University ; Humans ; Malaysia ; Male ; Middle Aged ; Prospective Studies ; Thalassemia ; blood ; Transfusion Reaction