Chronic myeloid leukaemia (CML) provides an illustrative disease model for both molecular pathogenesis of cancer and rational drug therapy. Imatinib mesylate (IM), a BCR-ABL1 targeted tyrosine kinase inhibitor (TKI) drug, is the first line gold standard drug for CML treatment. Conventional cytogenetic analysis (CCA) can identify the standard and variant Philadelphia (Ph) chromosome, and any additional complex chromosome abnormalities at diagnosis as well as during treatment course. Fluorescence in situ hybridization (FISH) is especially important for cells of CML patients with inadequate or inferior quality metaphases or those with variant Ph translocations. CCA in conjunction with FISH can serve as powerful tools in all phases of CML including the diagnosis, prognosis, risk stratification and monitoring of cytogenetic responses to treatment. Molecular techniques such as reverse transcriptase-polymerase chain reaction (RT-PCR) is used for the detection of BCR-ABL1 transcripts at diagnosis whereas quantitative reverse transcriptase-polymerase chain reaction (qRTPCR) is used at the time of diagnosis as well as during TKI therapy for the quantitation of BCR-ABL1 transcripts to evaluate the molecular response and minimal residual disease (MRD). Despite the excellent treatment results obtained after the introduction of TKI drugs, especially Imatinib mesylate (IM), resistance to TKIs develops in approximately 35% - 40% of CML patients on TKI therapy. Since point mutations in BCR-ABL1 are a common cause of IM resistance, mutation analysis is important in IM resistant patients. Mutations are reliably detected by nested PCR amplification of the translocated ABL1 kinase domain followed by direct sequencing of the entire amplified kinase domain. The objective of this review is to highlight the importance of regular and timely CCA, FISH analysis and molecular testing in the diagnosis, prognosis, assessment of therapeutic efficacy, evaluation of MRD and in the detection of BCR-ABL1 kinase mutations which cause therapeutic resistance in adult CML patients.

The long non-coding RNAs (lncRNAs) are the most prevalent and functionally diverse member of the non-coding RNA (ncRNA). The lncRNA has previously been considered to be a form of transcriptional “noise” but recent studies have found that the lncRNA to be associated with various disease conditions. It has also been found to play important roles in various physiological processes such as haemopoiesis, where lncRNA is reported to act as a fine-tuner of this very important process. To date, the effects of dysregulated lncRNA in thalassaemia has not been fully explored. This review article focuses on the possible roles of dysregulated lncRNAs in the pathogenesis of thalassaemia.

Interleukin-23 (IL-23) and IL-17 are the gatekeepers of CD4+ T helper 17 (Th17) cells where IL-23 is required for the development and expansion of Th17 cells that subsequently produce IL-17 to promote inflammation. Owing to such pro-inflammatory properties, the IL-23/IL-17 axis has emerged as an important mechanism in the pathogenesis of autoimmune diseases including systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA). In recent years, therapeutic antibodies targeting IL-23 (e.g. ustekinumab, tildrakizumab, guselkumab) or IL-17 (e.g. brodalumab, secukinumab, ixekizumab) have been approved for the treatment of various autoimmune diseases. In this review, we describe the pathogenic mechanisms of IL-23/IL-17 axis in SLE and RA, as well as summarising the findings from phase II and III clinical trials of anti-IL-23/IL-17 therapeutic antibodies in SLE and RA patients. In particular, phase II study has demonstrated that the anti-IL-23 antibody (ustekinumab) confers enhanced treatment outcomes in SLE patients, while anti-IL-17 antibodies (secukinumab and ixekizumab) have shown improved clinical benefits for RA patients in phase II/III studies. Our review highlights the emerging importance of targeting the IL-23/IL-17 axis in SLE and RA patients

The coronavirus disease-19 (COVID-19) has become a global pandemic of acute respiratory disease in just less than a year by the middle of 2020. This disease caused by the severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2), has resulted in significant mortality especially among the older age population and those with health co-morbidities. In contrast, children are relatively spared of this potentially ravaging disease that culminates in the acute respiratory distress syndrome, multi-organ failure and death. SARS-CoV-2 infection induces exuberant release of pro-inflammatory mediators, causing a “cytokine storm” and hypercoagulable states that underlie these complications. The SARS-CoV-2 infection median incubation is 5.1 days, with most developing symptoms by 11.5 days. It is highly infectious, spreading via the horizontal mode of transmission, but there is yet very limited evidence of vertical transmission to the newborn infant occurring either transplacentally or through breastfeeding. This said, various immune factors during childhood may modulate the expression of COVID-19, with the multisystem inflammatory syndrome in children (MIS-C) at the severe end of the disease spectrum. This article gives an overview of the SARS-CoV-2 infection, clinical presentation and laboratory tests of COVID-19 and correlating with the current understanding of the pathological basis of this disease in the paediatric population.

In 2003, it was discovered that the entry receptor for the Severe Acute Respiratory Syndrome coronavirus (SARS-CoV) is a protein called the angiotensin-converting enzyme 2 (ACE2). This protein is present in a number of cell types, including those from the respiratory tract. Soon after the emergence of SARS-CoV-2 that is responsible for the disease Covid-19, scientists found that ACE2 was also used by the new coronavirus to infect cells. This opened some interesting possibilities to explain the striking variation in risks of catching and dying from Covid-19. The best recognised of these are the much higher risk of serious illness in older than younger people, in men than women, and in those with pre-existing comorbidities such as hypertension and cardiovascular diseases. There are several ways in which the ACE2 protein might contribute to this variation. The most obvious would be if there is more ACE2, there would be more entry points for the virus to infect the cell, e.g. in older people or in men. However, the evidence for this is rather small, partly because it is not that easy to obtain representative healthy tissues. Alternatively, it could be related to ACE2 membership of a family of proteins that has one end of the protein anchored inside the cell while most of the protein protrudes from the outside of the cell which therefore can be shed when cleaved by proteases at the cell membrane. Herein we review current evidence and theories of ACE2 role on SARS-CoV-2 infectivity and Covid-19 severity.

Together with isocitrate dehydrogenase (IDH) mutation, co-deletion of 1p19q (1p19q codel) is a prerequisite for diagnosis of oligodendroglioma, making it imperative that histopathology laboratories introduce testing for 1p19q codel. To date there is still no consensus reference range and cut-offs that confirm deletion of 1p or 19q. We embarked on determining our reference range in 11 formalinfixed, paraffin-embedded non-neoplastic brain tissue using fluorescence in situ hybridisation (FISH) with the Vysis 1p36/1q25 and 19q13/19p13 FISH Probe Kit (Abbott Molecular Inc., USA). At same time we attempted to validate our methodology in 13 histologically-confirmed IDH-mutant oligodendrogliomas. For 1p, percentage cells with deletion (range=8-23%; mean±SD = 15.73±5.50%) and target: control (1p36:1q25) ratio (range = 0.89-0.96; mean±SD = 0.92±0.03) in non-neoplastic brain, differed significantly (p<0.000) from oligodendroglioma (percentage cells with deletion: range = 49-100%; mean±SD = 82.46±15.21%; target:control ratio range:0.50-0.76; mean±SD = 0.59±0.08). For 19q, percentage cells with deletion (range = 7-20%; mean±SD = 12.00±3.49%) and target:control (19q13/19p13) ratio (range:0.90-0.97; mean±SD = 0.94±0.02) in non-neoplastic brain also differed significantly from oligodendroglioma (percentage cells with deletion: range = 45-100%; mean±SD = 82.62±18.13%; target:control ratio range:0.50-0.78; mean±SD = 0.59±0.09). Using recommended calculation method, for diagnosis of 1p deletion, percentage of cells showing deletion should be >32-33% and/or target:control ratio <0.83. For 19q, percentage of cells showing deletion should be >22% and target:control ratio <0.88. Using these cut-offs all 13 oligodendroglioma demonstrated 1p19q codel.

Introduction: Recent studies have published the roles of exosomal miRNAs in the pathogenesis of various type of malignancies and can be developed as potential biomarkers for diagnostic, prognostic and therapeutic purposes. The aim of this study was to identify the expression level of selected miRNAs (miR-182, miR-301a, and miR-373) in exosomes of the serum and ascitic fluid in patients with non-alcoholic steatohepatitis (NASH)-related liver cirrhosis with or without hepatocellular carcinoma (HCC). Materials and Methods: A literature search was performed to identify potential miRNAs involved in the pathogenesis of HCC. Unpaired serum and ascitic fluid were obtained from 52 patients with NASH related liver cirrhosis (n=26 for each group of with and without HCC). Exosomal miRNA was isolated from all samples. Expression levels of miR-182, miR-301a and miR373 were determined using quantitative real-time PCR. Results: Serum-derived exosomal mir-182, miR-301a and miR-373 were significantly up-regulated with fold change of 1.77, 2.52, and 1.67 (p< 0.05) respectively in NASH-induced liver cirrhosis with HCC as compared to NASH-induced liver cirrhosis without HCC. We identified the expression levels of ascitic fluid-derived exosomal mir-182, miR-301a, and miR-373 were significantly up-regulated with fold change of 1.6, 1.94 and 2.13 respectively in NASH-induced liver cirrhosis with HCC as compared to NASH-induced liver cirrhosis without HCC (p <0.05). There was poor correlation expression of all the selected exosomal miRNA between serum- and ascitic fluid-derived in HCC group. Conclusions: This preliminary data showed significant increase in the expression levels of exosomal miR-182, miR-301a and miR373 in both serum and ascetic fluid suggesting the possible roles of these miRNAs as circulating biomarkers for NASH-induced liver cirrhosis with hepatocellular carcinoma

Introduction: Ionised calcium is a good prognostic and diagnostic tool as opposed to total calcium in critical patients but is not available in most central laboratories and non-intensive care units. To date, four equations to calculate ionised calcium in critical patients have been published. Objectives: (1) Evaluate the four published equations’ performance in estimating ionised calcium; (2) Determine the accuracy of calculated ionised and adjusted total calcium in classifying patients according to calcium states; and (3) Identify factors associated with hypocalcaemia in the critically ill population. Materials and methods: This is a cross-sectional study involving 281 critically ill patients aged 18-80 years of both genders in a Malaysian tertiary intensive care unit. Performance of the four equations was analysed using Bland-Altman difference plot and Passing Bablok regression analysis. Crosstabulation was conducted to assess classification accuracy. Mann-Whitney U or Pearson Chi-Square tests were performed to identify variables associated with hypocalcaemia. Results: Calculated ionised calcium using all four equations significantly overestimated ionised calcium. Calculated ionised and adjusted total calcium had poor accuracies in classifying hypocalcaemic patients. pH was significantly higher in hypocalcaemics. Conclusion: Calculated ionised and adjusted total calcium significantly overestimate ionised calcium in the critically ill. In this specific population, calcium status should only be confirmed with ionised calcium measured by direct ion-selective electrode (ISE).

Introduction: One commonly used equation which continues to be widely mentioned in text books and hence familiar to clinical people is total calcium + 0.02 (40 – albumin). This equation was derived using cresophthalein complexone and bromocresol green (BCG) methods for measuring serum total calcium and serum albumin respectively. However this equation maybe invalid when applied to calcium and albumin results generated by alternative assays. Hence we aim to derive an albumin-adjusted calcium equation specific to our laboratory’s total calcium and albumin methodologies. Materials and Methods: A total of 3,175 adult University Malaya Medical Centre (UMMC) patients deemed free of any calcium metabolism disorders were selected and divided into two groups for derivation and validation. Simple linear regression associating total calcium and albumin was constructed from the data in the derivation group. The new albumin-adjusted calcium equation was validated in the validation group. Differences in calcium status classification following adjustments based on existing and new albumin-adjusted calcium equation was compared in a 469 hypoalbuminaemic patients. Result: The new albumin adjusted calcium equation was: total calcium + 0.014 x (39-albumin). Of the 469 hypoalbuminemic patients, 78 were classified differently based on new equation. Based on the new equation, 55 normocalcemic patients were classified as hypocalcemic and 22 were classified as normocalcemic instead of hyperclacaemic. Conclusion: Based on the newly derived albuminadjusted calcium equation 17% of patients had different adjusted calcium classifications. This could potentially impact in the management. It is recommended that laboratories derive equations specific to their calcium/albumin methods and analytical platforms.

Introduction: Rifampicin is a key first-line antimycobacterial agent employed for the treatment of pulmonary tuberculosis (PTB). This study sought to obtain prevalence data on rifampicin-resistant Mycobacterium tuberculosis among smear-positive PTB patients in the Klang District of Malaysia. Materials and Methods: A total of 103 patients from the Chest Clinic of Hospital Tengku Ampuan Rahimah with sputum smears positive for acid-fast bacilli were included in this cross-sectional study. All sputa were tested using Xpert MTB/RIF to confirm the presence of M. tuberculosis complex and detect rifampicin resistance. Sputa were also sent to a respiratory medicine institute for mycobacterial culture. Positive cultures were then submitted to a reference laboratory, where isolates identified as M. tuberculosis complex underwent drug susceptibility testing (DST). Results: A total of 58 (56.3%) patients were newly diagnosed and 45 (43.7%) patients were previously treated. Xpert MTB/RIF was able to detect rifampicin resistance with a sensitivity and specificity of 87.5% and 98.9%, respectively. Assuming that a single resistant result from Xpert MTB/RIF or any DST method was sufficient to denote resistance, a total of 8/103 patients had rifampicinresistant M. tuberculosis. All eight patients were previously treated for PTB (p<0.05). The overall prevalence of rifampicin resistance among smear-positive PTB patients was 7.8%, although it was 17.8% among the previously treated ones. Conclusion: The local prevalence of rifampicin-resistant M. tuberculosis was particularly high among previously treated patients. Xpert MTB/RIF can be employed in urban district health facilities not only to diagnose PTB in smear-positive patients, but also to detect rifampicin resistance with good sensitivity and specificity.