Journal of Haematology and Stem Cell Research https://jhscr.org/index.php/JHSCR <p><em>Journal of Haematology and Stem Cell Research (JHSCR)</em> is an official journal of Pakistan Society of Haematology, recognized by <strong>Pakistan Medical and Dental Council IP/0084 till September 2025</strong> &amp; Higher Education commission of Pakistan in Y category ). It is a double blind double peer-reviewed being published biannual journal that delivers the premier quality peer-reviewed open access original research articles, reviews, and scholarly comment on pioneering efforts and innovative studies in the medical treatment of blood disorders. The journal has a strong clinical and pharmacological focus and is aimed at an international audience of clinicians and researchers in hematology and related disciplines, providing an online forum for rapid dissemination of recent research and perspectives in this area.</p> Pakistan Society of Haematology en-US Journal of Haematology and Stem Cell Research 2790-2277 <p><a href="http://creativecommons.org/licenses/by-nc/4.0/" rel="license"><img src="https://i.creativecommons.org/l/by-nc/4.0/88x31.png" alt="Creative Commons License" /></a><br />This work is licensed under a <a href="http://creativecommons.org/licenses/by-nc/4.0/" rel="license">Creative Commons Attribution-NonCommercial 4.0 International License</a>.<br />The licensor permits others to copy, distribute, display, and perform the work, as well as make and distribute derivative works based on it. The licensor permits others to copy, distribute, display, and perform the work for non-commercial purposes only.</p> Haematology Analyzers- Something More Than the Simple Blood Cells https://jhscr.org/index.php/JHSCR/article/view/183 Nadeem Ikram Copyright (c) 2025 Journal of Haematology and Stem Cell Research https://creativecommons.org/licenses/by-nc/4.0 2025-01-22 2025-01-22 4 2 220 224 Mutations at exons 11 and 17 of c-kit Gene in Patients of Acute Myeloid Leukaemia https://jhscr.org/index.php/JHSCR/article/view/126 <p><strong>Objective:</strong> To determine the frequency of mutations at exons 11 and 17 of c-kit gene in patients of Acute Myeloid Leukaemia.</p> <p><strong>Methodology:</strong> It was descriptive study and conducted in departments of Hematology and Human Genetics and Molecular Biology, University of Health Sciences, Lahore. Total 80 newly diagnosed patients of AML from different hospitals of Lahore were included in the study The study included both peripheral blood and bone marrow aspirate slides of 80 newly diagnosed patients of AML. Each sample was amplified for exons 11 and 17 of c-kit gene using specific primers by direct DNA sequencing.</p> <p><strong>Results:</strong> Among 80 patients, 67.5% (n = 54) were males and 32.5% (n = 26) were females with a median age of 38 years (range 12-85 years). Our DNA sequencing results indicated that none of the patient had been observed to have mutation in both exons of c-kit gene. Only a polymorphism was identified in exon 11. So, it is concluded that our screened AML population was negative for these two exons of c-kit gene.</p> <p><strong>Conclusion</strong>: The study concluded that our selected AML patients were negative for these two exons but SNP was found in exon 11 of c-kit gene.</p> Yusra Rashid Moizza Sahar Ghulam Mustafa Muhammad Asif Naveed Shahida Mohsin Shagufta Khaliq Copyright (c) 2025 Journal of Haematology and Stem Cell Research https://creativecommons.org/licenses/by-nc/4.0 2025-01-22 2025-01-22 4 2 225 229 Frequency of FLT3–ITD Mutation in Newly Diagnosed Acute Myeloid Leukemia: A Single-Center Study in Pakistan https://jhscr.org/index.php/JHSCR/article/view/184 <p><strong>Objectives: </strong>To determine the frequency of FLT3-ITD mutation in newly diagnosed cases of acute myeloid leukemia.&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</p> <p><strong>Methodology: </strong>The cross sectional study was carried out over a period of one year from January to December in the department of hematology, Armed Forces Institute of Pathology (AFIP), Rawalpindi Total 85 patients were included in this study. After obtaining 3 ml of venous blood in EDTA tubes, DNA was extracted using QlAamp DNA kit. Conventional PCR was performed for detection of FLT3-ITD mutation with positive and negative controls.</p> <p><strong>Results</strong>: FLT3-ITD mutations were detected in 15 cases (17.6%). Out of 85 cases, 56 (65.9%) were males while remaining 29 patients (34.1%) were females. Most common AML type according to French American British (FAB) classification was AML-M2. 33 cases (38.8%). Mean values of different hematological variables were TLC 51.8 x10<sup>9</sup>/L (±80.98), Hb9.16 g/dl (±8.15), Platelets 59.25 x10<sup>9</sup>/L (±60.49) and Myeloblasts 47.57% (±27.93) in bone marrow aspirates.</p> <p><strong>Conclusion</strong>: It is concluded that FLT3-ITD mutations are quite frequent in our AML patients with different age specificity. Frequency of FLT3-ITD mutation in our study was lower (17.6%) than other international studies. The detection of these mutations would be helpful in risk stratification and selection of appropriate management of the patients.</p> Maliha Saad Ayesha Sarwar Muhammad Ashraf Shehla Alizai Maryam Zulfiqar Fareeha Shahid Copyright (c) 2025 Journal of Haematology and Stem Cell Research https://creativecommons.org/licenses/by-nc/4.0 2025-01-22 2025-01-22 4 2 230 233 SYBR Green PCR is Cost-Effective for Detecting Genetic Abnormalities in Acute Myeloid Leukemia https://jhscr.org/index.php/JHSCR/article/view/132 <p><strong>Objective:</strong> To determine whether SYBR Green PCR is cost-effective for detecting genetic abnormalities in acute myeloid Leukemia or not.</p> <p><strong>Methodology:</strong> This cross-sectional study was conducted at Department of Pathology-CMH Lahore medical college and institute of dentistry from October 2023- March 2024. DNA samples from 70 adults diagnosed as AML at various hospitals were ran on RT-PCR using dye and probe-based methods for detection of genetic abnormalities in acute myeloid leukemia (AML). Data was analyzed by SPSS 23 and presented in frequencies and percentages.</p> <p><strong>Results:</strong> In a total of 70 patients, twenty-five (17.5%) had defining genetic abnormalities. Fusions were detected in 19 (13.3%) patients while three rearrangements (2.1%) and three mutations (2.1%) were detected. The probe-based RT-PCR matched results in all patients.</p> <p>C<strong>onclusion</strong>: SYBR Green PCR is very cost-effective for detecting genetic abnormalities in acute myeloid leukemia.</p> Mohammad Abdul Naeem Farhat Ijaz Mohammad Amir Rana Khurram Aftab Copyright (c) 2025 Journal of Haematology and Stem Cell Research https://creativecommons.org/licenses/by-nc/4.0 2025-01-22 2025-01-22 4 2 234 237 Frequency of TP53 (17p13) Deletion in Patients with Chronic Lymphocytic Leukemia (CLL) https://jhscr.org/index.php/JHSCR/article/view/160 <p><strong>Objective: </strong>To determine the frequency of TP53 (17p13) gene deletion in patients diagnosed with chronic lymphocytic leukemia (CLL) and assess its correlation with age, gender, and total leukocyte count (TLC).</p> <p><strong>Methodology: </strong>This retrospective cross-sectional study was conducted at the Section of Hematology and Cytogenetics, Department of Pathology, Shaukat Khanum Memorial Cancer Hospital and Research Center, Lahore, from January 2019 to December 2021. A total of 157 CLL patients were included. Peripheral blood or bone marrow samples were analyzed using the Fluorescence In Situ Hybridization (FISH) technique to detect TP53 deletion. Statistical analysis was performed using SPSS version 21.0, and Chi-square tests were applied for significance assessment.</p> <p><strong>Results: </strong>Among the 157 CLL patients, 114 (72.61%) were males, and 43 (27.38%) were females, with a mean age of 56.44 ± 8.20 years. TP53 deletion was observed in 30 (19.11%) patients, including 23 (76%) males and 7 (23.30%) females. No significant correlation was found between TP53 deletion and age or gender (p &gt; 0.05). Additionally, 18 (60%) patients with TP53 deletion had TLC &lt;100×10³, while 12 (40%) had TLC &gt;100×10³.</p> <p><strong>Conclusion: </strong>The frequency of TP53 gene deletion in CLL patients was 19.11%. The deletion was more prevalent in males, though statistical significance was not established. Early identification of TP53 deletion is crucial for risk stratification and treatment decisions</p> Natalia Ahmad Ammarah Tahir Asad Hayat Ahmad Hiba Asif Mohammad Tariq Mehmood Kiran Imran Copyright (c) 2025 Journal of Haematology and Stem Cell Research https://creativecommons.org/licenses/by-nc/4.0 2025-01-22 2025-01-22 4 2 238 241 Determining the Correlation Between Automated vs. Manual Method for Reticulocyte Estimation https://jhscr.org/index.php/JHSCR/article/view/130 <p><strong>Objective: </strong>To determine the correlation between automated and manual methods for the determination of reticulocyte (Retic) count in symptomatic patients.</p> <p><strong>Methodology: </strong>This cross-sectional study was conducted at Rehman Medical Institute, Peshawar, from November 2021 to April 2022. A total of 384 patients of both genders and all ages were enrolled. Anemic patients with high Retic counts who were referred to RMI for reticulocyte (RC) examination were included. Samples that were improperly proportioned with anticoagulants, lipemic, hyperbilirubinemic, or hemolyzed were excluded. The reticulocyte counts of all samples were analyzed using both manual methods and an automated analyzer (Sysmex XN analyzer), with evaluations performed by two different hematologists to eliminate observer bias.</p> <p><strong>Results: </strong>No significant deviation was observed between manual and automated reticulocyte counts in male or female specimens (P &gt; 0.05). Similarly, there was no significant difference in average reticulocyte counts among adults, children, and infants (P &gt; 0.05). However, a noticeable difference in mean manual and automated RC was observed across different types of anemia, including microcytic, normocytic, and macrocytic anemia. Additionally, the mean reticulocyte hemoglobin cellular content, reticulocyte volume, and immature reticulocyte fraction showed significant differences across all specimens (P &lt; 0.001).</p> <p><strong>Conclusion: </strong>There were no significant deviations in reticulocyte counts between the manual and automated methods. The manual method is preferred for its cost-effectiveness, while the automated approach is recommended for its efficiency and accuracy.</p> Iqra Zeb qureshi Hina mushtaq Hamid Iqbal Marium Raza Fuad Ahmad Siddiqui Imran Khan Copyright (c) 2025 Journal of Haematology and Stem Cell Research https://creativecommons.org/licenses/by-nc/4.0 2025-01-22 2025-01-22 4 2 242 246 Association of Ferritin Levels and Biochemical Markers with COVID-19 Severity https://jhscr.org/index.php/JHSCR/article/view/185 <p><strong>Objective:</strong> To investigate the association between ferritin levels and other biochemical markers (such as total bilirubin, LDH, AST, and ALT) with the severity of COVID-19 and to identify potential predictive markers that could help in assessing disease severity, improving patient management, and reducing mortality rates.</p> <p><strong>Methodology:</strong> This retrospective observational study was conducted between June 2020 to December 2020 admitted to various hospitals in twin cities, comprised 420 confirmed cases of COVID-19 detected through PCR testing. Blood samples collected upon admission from patients in various hospitals across Islamabad and Rawalpindi were analyzed for Lactate Dehydrogenase (LDH), Aspartate Aminotransferase (AST), Alanine Transaminase (ALT), Total Bilirubin and ferritin levels . The diagnosis was made using Reverse Transcription-Polymerase Chain Reaction (RT-PCR) on nasal and pharyngeal swab specimens</p> <p>The data was statistically analyzed by using SPSS ver 25. The Kruskal-Wallace test was used to compare severe, moderate, and mild patient COVID-19 groups</p> <p><strong>Results:</strong> The mean age of the deceased was significantly high 76.09±?4.94 years vs 54.34?±?13.03 years compared to those recovered. Deceased had ten times higher median levels of ferritin 1000 ng/ml vs. 120ng/ml. In multivariate analysis, advanced age (AOR 1.673 C.I (1.372,2.040) p = 0.0001)and a raised level of ferritin (AOR 2.938 C.I. (1.244,6.941) p = 0.014) showed significant association with mortality.</p> <p><strong>Conclusion:</strong> Ferritin and age can be used to predict the severity of COVID-19. This study can help physicians to lower the mortality rate and number of admissions in the ICU.</p> Syeda Saman Batool Feroza Hamid Watto Matloob Rana Sarah Naheed Sadaf Sultana Rizwan Ahmed Kiani Nadeem Ikram Copyright (c) 2025 Journal of Haematology and Stem Cell Research https://creativecommons.org/licenses/by-nc/4.0 2025-01-22 2025-01-22 4 2 247 251 Association of Iron Overload with Insulin Resistance in Transfusion-Dependent Beta Thalassemia Major Patients via Homeostatic Beta-Cell Model (HOMA-Beta) and Homeostatic Insulin Resistance Model (HOMA-IR): Insights from a Thalassemia Center in Karachi, Pakistan https://jhscr.org/index.php/JHSCR/article/view/148 <p><strong>Objective:</strong> To determine incidence of diabetes mellitus in transfusion dependent beta-thalassemia major patients, pancreatic beta-cell function by HOMA-beta &amp; insulin resistance by HOMA-IR in beta-thalassemia major patients and the relation of beta-cell function and insulin resistance with iron overload in transfusion dependent beta-thalassemia major patients.</p> <p>Methodology: This cross-sectional study was conducted in Baqai Medical University Karachi and Muhammadi Thalassemia Center Karachi. A total of 100 patients of both genders; age group between 2 years to 16 years were included in this study. Lab investigations were performed for FBS, Plasma Insulin, HbA1c, and Serum Ferritin on fully automated chemiluminence analyzer. HOMA-IR and HOMA-Beta were calculated.</p> <p><strong>Results:</strong> Prevalence of diabetes mellitus in transfusion dependent beta-thalassemia major patients was 15%. According to HOMA-IR, 48% patients had severe insulin resistance. HOMA-Beta was significantly lower in diabetic than non-diabetic patients. Relationship between HOMA-IR and HOMA-Beta according to iron overload status (? 2500 and &gt;2500) was positive but the correlation was strong (r=0.85) between HOMA-IR and HOMA-Beta in patients whose iron overload status was less than and equal to 2500 however correlation of these HOMA-IR and HOMA-Beta was week (r=0.209) in those whose iron overload was &gt;2500.</p> <p><strong>Conclusion:</strong> Iron overload is significantly promoting insulin resistance in transfusion-dependent beta-thalassemia major patients. Beta-thalassemia major patients are developing insulin resistance because of repeated blood transfusions. HOMA-Beta and HOMA-IR are reliable predictors of insulin resistance.</p> Syeda Hira Abid Muhammad Younus Jamal Maeesa Sajeel Sadia Talib Zarrish Qasim Naheed Akhtar Copyright (c) 2025 Journal of Haematology and Stem Cell Research https://creativecommons.org/licenses/by-nc/4.0 2025-01-22 2025-01-22 4 2 252 256 Investigating the Influence of Pre-Donation Haematological Parameters on Haematocrit and the Haemoglobin Content in Packed Red Blood Cell Units: A Single Centre Study https://jhscr.org/index.php/JHSCR/article/view/149 <p><strong>Objective:</strong> This study aimed to evaluate the impact of pre-donation hematological parameters, specifically hemoglobin and hematocrit levels, on the quality of packed red blood cell (PRBC) units after 21 days of storage.</p> <p><strong>Methodology:</strong> A cross-sectional, analytical study was conducted at the Armed Forces Institute of Transfusion, Rawalpindi, involving 384 voluntary whole blood donors who met specific inclusion criteria. Pre-donation hematological parameters were measured using an automated hematology analyzer. PRBC units were processed according to standard protocols and evaluated on Day 21 for volume, hematocrit, and total hemoglobin content. Data analysis was performed using SPSS software, applying descriptive statistics, Pearson correlation, and multiple regression analysis to assess the relationship between pre-donation variables and PRBC quality.</p> <p><strong>Results:</strong> At Day 21, PRBC units exhibited significant changes compared to initial post-donation values. Pre-donation hemoglobin and hematocrit levels showed weak correlations with Day 21 PRBC hemoglobin content and hematocrit levels. The integrity of PRBC units appeared more influenced by storage conditions than donor-specific hematological parameters. While hemoglobin content remained relatively stable, hematocrit levels showed variability, indicating that additional biochemical and environmental factors during storage play a critical role in maintaining PRBC quality.</p> <p><strong>Conclusion:</strong> Pre-donation hematological parameters have a limited role in predicting PRBC quality after 21 days of storage. Factors such as donor conditions and processing parameters may significantly influence PRBC integrity, highlighting the need for comprehensive donor screening and optimized storage practices.</p> Mahnoor Siddiqui Umbreen Hashim Fahim Akhtar Rehan Lodhi Sadia Ali Minun Siddiqui Fuad Ahmad Siddiqui Copyright (c) 2025 Journal of Haematology and Stem Cell Research https://creativecommons.org/licenses/by-nc/4.0 2025-01-22 2025-01-22 4 2 257 263 The Detection of Beta Thalassemia Trait in Siblings of Beta Thalassemia Major Patients in Karachi: A Step towards Beta Thalassemia Major Free Society https://jhscr.org/index.php/JHSCR/article/view/155 <p><strong>Objective:</strong>&nbsp; To detect ?-Thalassemia trait in siblings of ?-Thalassemia major patients in Karachi</p> <p><strong>Background:</strong> Beta thalassemia major (BTM) is diagnosed very early in life and the patient usually has signs of hemolysis as well 7. Beta-thalassemia minor is another form of beta-thalassemia, in which the patient is usually asymptomatic and shows only mild anemia on CBC labs 12. Being more precise, if both partners carry beta thalassemia trait, then there is 25% chance of having homozygous beta thalassemia offspring. Considering this factor, the present study is designed to identify the presence of thalassemia minor in the siblings of Beta-thalassemia major patients.</p> <p><strong>Materials and Methodology: </strong>This cross-sectional study was conducted at Muhammadi Institute of Hematology in Karachi. Blood samples were collected for complete blood count and high-performance liquid chromatography. Data was analyzed using SPSS with comparison of parameters between non-carrier and carrier group done using independent t-test (P&lt;0.05).</p> <p><strong>Results:</strong> From the 400 siblings screened, 168 (42%) were found to be carriers of the beta thalassemia trait. On CBC interpretation, those who were carriers showed a significant reduction in MCV and MCH, where as their RBC count was high and MCHC was normal. The HbA, HbF, and Hb A2 levels were also significantly different between the carrier group and non-carrier group.</p> <p><strong>Conclusion:</strong> Prevalence rate of beta thalassemia was higher in the siblings of beta-thalassemia major patients. Therefore, proper screening, identification, and genetic counseling in this group can be extremely valuable</p> Ghazal Irfan Ghazal Maeesa Wadood maeesa Sarah Azhar sarah Muhammad khan Khan Ruqaya Nangrejo Tooba khan Yasir Rishi yasir Copyright (c) 2025 Journal of Haematology and Stem Cell Research https://creativecommons.org/licenses/by-nc/4.0 2025-02-07 2025-02-07 4 2 Utilizing WBC Differential Channel and Abnormal WBC Flags for Diagnosing Hematological Malignancies Using Sysmex XN-9000 Analyzer https://jhscr.org/index.php/JHSCR/article/view/139 <p><strong>Background: </strong>The haematology analyser, Sysmex XN-9000, generates white blood cell count with varying scattering intensities and abnormal WBC flags like monocytosis/abnormal lymphocytes/blasts during a complete blood count (CBC) analysis. The objective of the study is to determine the predictive role of abnormal flags (monocytosis; blast/abnormal lymphocytes; atypical lymphocytes) along with variation of WDF scatter plot and comparing them with peripheral blood smears for diagnosing and differentiation of leukaemia.</p> <p><strong> </strong><strong>Methodology: </strong>This study was performed at Chughtai Lab, Lahore from July, 2023 till October, 2023<strong>.</strong> A total of 160 EDTA samples were run on Sysmex NX 9000. Parameters that were studied and compared were WBC count, absolute monocyte count, monocyte percentage, abnormal flags and WDF scattergram. The results were confirmed by microscopy. Data was analyzed by SPSS version 23.00. Mean±SD was calculated for the continuous variable. Frequency and percentage were calculated for categorical variables. Pearson coefficient was performed to assess the correlation between various hematological parameters. Logistic regression was performed to determine the predictive role of abnormal flags and WDF scatter plot patterns in diagnosing leukemia. A p-value of &lt;0.05 was taken as significant</p> <p><strong>Results:</strong></p> <p>The CBC analysis by the instrument shows a good ability to detect true positive cases (sensitivity of 57.5%). Specifically, it has zero specificity and negative predictive value (NPV), meaning it cannot accurately identify true negatives, leading to a high false positive rate. Consequently, the instrument test cannot reliably rule out the condition when it gives a negative result, necessitating confirmatory testing with the gold standard i.e. microscopic examination of the peripheral smear.</p> <p><strong>Conclusion:</strong></p> <p>Relying solely on the flags generated by Sysmex XN 9000 does not warranty accurate diagnosis. Further, examining of the peripheral blood smear should be mandatory for the detection of suspicious cells whenever an abnormal flag is generated.</p> <p><strong>Key words</strong>: Automated hematology analyzer, Abnormal flags, Sysmex XN, Monocytosis, Atypical lymphocytes, WBC Flags</p> Muhammad Usman Siddique Aiman Mahmood Minhas Urooj Irfan Nimrah Ishaque Ayisha Imran Nomaan A. Malik Copyright (c) 2025 Journal of Haematology and Stem Cell Research https://creativecommons.org/licenses/by-nc/4.0 2025-01-22 2025-01-22 4 2 DETECTION OF IRON DEFICIENCY USING RETICULOCYTE HAEMOGLOBIN IN ADULT PATIENTS HAVING MICROCYTIC HYPOCHROMIC ANAEMIA WITH NORMAL OR RAISED SERUM FERRITIN https://jhscr.org/index.php/JHSCR/article/view/129 <p><strong>OBJECTIVE:</strong> To detect iron deficiency using reticulocyte haemoglobin in patients with microcytic hypochromic anemia having normal or raised serum ferritin.</p> <p><strong>STUDY DESIGN</strong>: Cross-sectional study</p> <p><strong>PLACE AND DURATION OF STUDY:</strong> This study was conducted in ddepartment of Haematology, Armed Forces Institute of Pathology (AFIP), Rawalpindi from February 2023 to July 2023.</p> <p><strong>METHODOLOGY: </strong>This cross-sectional study included 323 patients who presented with microcytic hypochromic anemia having normal or raised serum ferritin. Blood samples were taken in an Ethylenediamine tetra-acetic acid (EDTA) anticoagulant tubes and Sysmex XN 1000 analyzer was used to measure RBC parameters including reticulocyte Haemoglobin (Ret-He). Chi-square test and Independent T-test were applied for qualitative variables and quantitative variables respectively. Pearson’s correlation test was used to correlate reticulocyte hemoglobin with RBC parameters. P value &lt; 0.05 was considered statistically significant.</p> <p> <strong>RESULTS:</strong> Current study showed that the reticulocyte Haemoglobin with a cutoff value of 29 pg (sensitivity of 94.63% and a specificity of 85.48%, p&lt;0.0001) is an efficient and reliable indicator to detect iron deficiency in patients presenting with microcytic hypochromic blood picture with normal or raised serum ferritin (r = +0.217, p &lt; 0.001).</p> <p><strong>CONCLUSIONS:</strong> Our study concluded that reticulocyte haemoglobin can be a valuable marker for detecting iron deficiency in adult patients with microcytic hypochromic anaemia, especially when serum ferritin levels are normal or elevated.</p> Saima Iram Manzar Bozdar Hamid Saeed Malik Shazia Bano Rafia Mahmood Ayesha Khurshid Aysha Khan Sundas Durrani Copyright (c) 2025 Journal of Haematology and Stem Cell Research https://creativecommons.org/licenses/by-nc/4.0 2025-01-22 2025-01-22 4 2