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  1. Fulltext Intravenous iron-sucrose complex for treatment of iron deficiency anaemia in pregnancy - experience in an obstetric day care clinic in a tertiary hospital
    Tan, AE, Siti, S.A.
    Medicine & Health, 2008;3(2):288-293.
    MyJurnal
    A cross-sectional study was undertaken to evaluate if outpatient administration of in-travenous iron sucrose complex (Venofer) was a sensible option in treating iron defi-ciency anaemia during pregnancy and puerperium. A total of 120 patients with iron deficiency anaemia were recruited from the Obstetric Day Care Clinic at the Universiti Kebangsaan Malaysia Medical Centre (UKMMC) over 18 months from March 2003 to August 2004. The main outcome measures were haemoglobin increment, patients’ compliance, adverse effects and saving from hospitalization fees. The pre-treatment haemoglobin (Hb) level was 8.5+0.85g/dl for the antenatal patient and 7.6+0.80 g/dl in the post-partum group. The mean post-treatment haemoglobin increment at day four-teenth was 3.52+0.75g/dl. One patient developed skin rash while another had low-grade pyrexia. Seven patients experienced mild metallic taste. There were no serious side effects or anaphylactic reactions. Ten patients (8.3%) did not complete their ther-apy - eight delivered before completion of treatment; another two defaulted following delivery. The average number of Venofer used was seven ampoules i.e. 700mg per person, most of them required three sessions to complete the course. Outpatient treatment allows each patient to save hospitalization fees of RM45 per day, which to-talled up to RM135 for a 3-days ward stay. An estimation of RM16,200 hospitalization fees for the 120 patients was avoided during the study period. In conclusion, outpatient treatment of anaemia in pregnancy and post-partum period using Venofer was safe and feasible, with high patient compliant and cost-savings from hospitalization fees. 
    Study site: Obstetric Day Care Clinic, Pusat Perubatan Universiti Kebangsaan Malaysia (PPUKM), Kuala Lumpur, Malaysia
  2. Insulin-like growth factor-1 receptor expression in the placentae of diabetic and normal pregnancies
    Hayati AR, Cheah FC, Tan AE, Tan GC
    Early Hum Dev, 2007 Jan;83(1):41-6.
    PMID: 16750336 DOI: 10.1016/j.earlhumdev.2006.04.002
    BACKGROUND: Septal hypertrophic cardiomyopathy (sHCM) is a characteristic anomaly of the infant of diabetic mother (IDM). Insulin-like growth factor-1 (IGF-1) has been identified as a mediator of tissue overgrowth and we have previously shown that maternal IGF-1 levels were significantly elevated among neonates with asymmetrical sHCM. IGF-1 does not cross the placenta; it exerts physiologic action through binding to the IGF-1 receptor (IGF-1R). Localisation and expression of IGF-1R in term diabetic pregnancies are largely unclear. We have studied IGF-1R in the placentae of diabetic and normal pregnancies and this receptor expression in association with neonates with sHCM.
    METHODS: IGF-1R localization and expression in the placentae of six diabetic pregnancies associated with neonatal sHCM were compared with six each of randomly selected diabetic and normal pregnancies without neonatal sHCM by immunohistochemistry. The staining for IGF-1R in the deciduas, cytotrophoblasts, syncytiotrophoblasts and villous endothelium for these 18 samples were assessed and scored by two pathologists who were blinded to the respective diagnoses.
    RESULTS: Placental IGF-1R staining was negative in the villous endothelium for all three groups. IGF-1R staining was present in deciduas, cytotrophoblasts and syncytiotrophoblasts but the staining was weaker in the entire group of infants with sHCM compared to those without sHCM.
    CONCLUSIONS: IGF-1R is localized in all cell types of the placenta except in villous endothelium. Weaker placental IGF-1R staining in the placentae of diabetic pregnancies associated with sHCM suggests reduced expression of IGF-1R. This may be a down-regulatory response to elevated maternal IGF with neonatal sHCM outcome.
  3. Organotypic culture of human amnion cells in air-liquid interface as a potential substitute for skin regeneration
    Fatimah SS, Chua K, Tan GC, Azmi TI, Tan AE, Abdul Rahman H
    Cytotherapy, 2013 Aug;15(8):1030-41.
    PMID: 23830235 DOI: 10.1016/j.jcyt.2013.05.003
    The aim of the present study was to evaluate the effects of air-liquid interface on the differentiation potential of human amnion epithelial cells (HAECs) to skin-like substitute in organotypic culture.
  4. Effects of epidermal growth factor on the proliferation and cell cycle regulation of cultured human amnion epithelial cells
    Fatimah SS, Tan GC, Chua KH, Tan AE, Hayati AR
    J Biosci Bioeng, 2012 Aug;114(2):220-7.
    PMID: 22578596 DOI: 10.1016/j.jbiosc.2012.03.021
    Human amnion epithelial cells (HAECs) hold great promise in tissue engineering for regenerative medicine. Large numbers of HAECs are required for this purpose. Hence, exogenous growth factor is added to the culture medium to improve epithelial cells proliferation. The aim of the present study was to determine the effects of epidermal growth factor (EGF) on the proliferation and cell cycle regulation of cultured HAECs. HAECs at P1 were cultured for 7 days in medium containing an equal volume mix of HAM's F12: Dulbecco's Modified Eagles Medium (1:1) supplemented with different concentrations of EGF (0, 5, 10, 20, 30 and 50 ng/ml EGF) in reduced serum. Morphology, growth kinetics and cell cycle analysis using flow cytometry were assessed. Quantitative gene expression for cell cycle control genes, pluripotent transcription factors, epithelial genes and neuronal genes were also determined. EGF enhanced HAECs proliferation with optimal concentration at 10 ng/ml EGF. EGF significantly increased the proportion of HAECs at S- and G2/M-phase of the cell cycle compared to the control. At the end of culture, HAECs remained as diploid cells under cell cycle analysis. EGF significantly decreased the mRNA expression of p21, pRb, p53 and GADD45 in cultured HAECs. EGF also significantly decreased the pluripotent genes expression: Oct-3/4, Sox2 and Nanog; epithelial genes expression: CK14, p63, CK1 and Involucrin; and neuronal gene expression: NSE, NF-M and MAP 2. The results suggested that EGF is a strong mitogen that promotes the proliferation of HAECs through cell cycle regulation. EGF did not promote HAECs differentiation or pluripotent genes expression.
  5. Value of human amniotic epithelial cells in tissue engineering for cornea
    Fatimah SS, Ng SL, Chua KH, Hayati AR, Tan AE, Tan GC
    Hum. Cell, 2010 Nov;23(4):141-51.
    PMID: 21166885 DOI: 10.1111/j.1749-0774.2010.00096.x
    Human amniotic epithelial cells (hAECs) are potentially one of the key players in tissue engineering due to their easy availability. The aim of the present study was to develop an optimal isolation and transportation technique, as well as to determine the immunophenotype and epithelial gene expression of hAECs. Amnion was mechanically peeled off from the chorion and digested with trypsin-ethylenediaminetetraacetic acid. The isolated hAECs were cultured in medium containing 10 ng/mL epidermal growth factor until P4. The epithelial gene expression, cell surface antigen and protein expression of hAECs were analyzed by quantitative polymerase chain reaction, flow cytometry and immunocytochemistry. hAECs were also cultured in adipogenic, osteogenic and neurogenic induction media. The best cell yield of hAECs was seen in the digestion of 15 pieces of amnion (2 × 2 cm) and isolated 30 min after digestion with trypsin. F12:Dulbecco's modified eagle medium was the best medium for short term storage at 4 °C. hAECs expressed CD9, CD44, CD73 and CD90, and negligibly expressed CD31, CD34, CD45 and CD117. After serial passage, CK3, CK19 and involucrin gene expressions were upregulated, while p63, CK1 and CK14 gene expressions were downregulated. Sustained gene expressions of integrin β1 and CK18 were observed. At initial culture, these cells might have stem-like properties. However, they differentiated after serial passage. Nonetheless, hAECs have epithelial stem cell characteristics and have the potential to differentiate into corneal epithelial cells. Further investigations are still needed to elucidate the mechanism of differentiation involved and to optimize the culture condition for long term in vitro culture.
  6. Human chorion-derived stem cells: changes in stem cell properties during serial passage
    Fariha MM, Chua KH, Tan GC, Tan AE, Hayati AR
    Cytotherapy, 2011 May;13(5):582-93.
    PMID: 21231803 DOI: 10.3109/14653249.2010.549121
    BACKGROUND AIMS: Fetal membrane from human placenta tissue has been described as a potential source of stem cells. Despite abundant literature on amnion stem cells, there are limited studies on the stem cell properties of chorion-derived stem cells.

    METHODS: The main aim was to determine the stemness properties of serial-passaged human chorion-derived stem cells (hCDSC). Quantitative polymerase chain reaction (PCR) was performed to reveal the following stemness gene expression in serial-passaged hCDSC: Oct-4, Sox-2, FGF-4, Rex-1, TERT, Nanog (3), Nestin, FZD-9, ABCG-2 and BST-1. Cell growth rate was evaluated from passage (P) 1 until P5. The colony-forming unit-fibroblast (CFU-F) frequency of P3 and P5 cells and multilineage differentiation potential of P5 cells were determined. The immunophenotype of hCDSC was compared using the surface markers CD9, CD31, CD34, CD44, CD45, CD73, CD90, CD117, HLA-ABC and HLA-DR, -DP and -DQ. Immunostaining for trophoblast markers was done on P0, P1, P3 and P5 cells to detect the contamination of trophoblasts in culture, while chromosomal abnormality was screened by cytogenetic analysis of P5 cells.

    RESULTS: The surface markers for mesenchymal lineage in hCDSC were more highly expressed at P5 compared with P3 and P0, indicating the increased purity of these stem cells after serial passage. Indeed, all the stemness genes except TERT were expressed at P1, P3 and P5 hCDSC. Furthermore, human chorion contained high clonogenic precursors with a 1:30 CFU-F frequency. Successful adipogenic, chondrogenic and osteogenic differentiation demonstrated the multilineage potential of hCDSC. The karyotyping analysis showed hCDSC maintained chromosomal stability after serial passage.

    CONCLUSIONS: hCDSC retain multipotent potential even at later passages, hence are a promising source for cell therapy in the future.

  7. Augmentation of angiogenic and endothelial associated gene expression by EDM50 in human chorion-derived stem cells
    Manzor NF, Chua KH, Tan GC, Tan AE, Abdul Rahman H
    Med J Malaysia, 2008 Jul;63 Suppl A:11-2.
    PMID: 19024960
    The objective of this study was to investigate the angiogenic potential of human chorion-derived stem cells (CDSC) cultured in medium containing bFGF and VEGF (EDM50). Total RNA was extracted from cells cultured in FD+10% FBS and EDM50. Quantitative RT-PCR was carried out to score the differential mRNA expression of genes involve in angiogenesis and endothelial differentiation. Our finding demonstrated that all angiogenic and endothelial associated genes were expressed higher in EDM50. Expression level of ANG-1, eNOS and VEGFR2 were significantly higher in EDM50 compared to FD+10% FBS. Our results suggested that human CDSC cultured in EDM50 can be used for angiogenesis purpose in regenerative medicine.
  8. The stemness gene expression of cultured human amniotic epithelial cells in serial passages
    Simat SF, Chua KH, Abdul Rahman H, Tan AE, Tan GC
    Med J Malaysia, 2008 Jul;63 Suppl A:53-4.
    PMID: 19024980
    The aim of the study is to evaluate the stemness gene expression of cultured human amniotic epithelial cells (HAECs) in serial passages. HAECs obtained from human term placentae were cultured in F12:DMEM(1:1) + 10% FBS +10ng/ml EGF in serial passages (P0, P1, P2 and P4). Quantitative RT-PCR was used to assess the gene expression analysis. The results showed that cultured HAECs expressed and downregulated the stemness genes expression for Oct-4, Sox-2, Nanog3, FGF4, Rex-1, FZD-9, BST-1 ABCG2. However, vimentin and nestin gene expression were upregulated. The results suggested that cultured HAECs may have pluripotent and multipotent properties.
  9. Quantitative RT PCR approach to evaluate the neurogenic and gliagenic gene expression of cultured human amniotic epithelial cells
    Tan GC, Simat SF, Abdul Rahman H, Tan AE, Chua KH
    Med J Malaysia, 2008 Jul;63 Suppl A:51-2.
    PMID: 19024979
    The aim of the study is to determine the neuronal and glial gene expression of cultured human amniotic epithelial cells (HAECs) in serial passages. HAECs obtained from human term placentae were cultured in F12:DMEM (1:1) + 10% FBS +10ng/ml EGF in serial passages. Quantitative RT-PCR was used to assess the gene expression analysis. The results showed that the cultured HAECs expressed the neural stem cell genes (Nestin, NSE and Vimentin), mature neuronal genes (TH, MAP-2, beta-III-tubulin and NFM) and glial genes (CNPase, MBP and Olig). These neural stem cell genes increased with serial passages while the genes expression for mature neuronal and glial cells were downregulated. These results suggested that HAECs may promote or involve in neurogenesis and gliagenesis.
  10. Upregulation of SOX-2, FZD9, Nestin, OCT-4 and FGF-4 expression in human chorion derived-stem cells after angiogenic induction
    Abdul Rahman H, Manzor NF, Tan GC, Tan AE, Chua KH
    Med J Malaysia, 2008 Jul;63 Suppl A:57-8.
    PMID: 19024982
    Angiogenic induction was made to promote angiogenesis by differentiating stem cells towards endothelial cells. However, the stemness property of induced cells has not been revealed yet. Hence, we aim to evaluate the differential mRNA expression of stemness genes in human chorion-derived stem cells (CDSC) after being cultured in EDM50 comprised bFGF and VEGF. Results indicated that CDSC cultured in EMD50 expressed significantly higher mRNA level of Sox-2, FZD9, BST-1 and Nestin. In addition Oct-4, FGF-4 and ABCG-2 were also upregulated. Our finding suggested that CDSC after angiogenic induction enhanced its stem cell properties. This could be contributed for the mechanism of stem cell therapy in ischemic problem.
  11. Laparoscopic salpingectomy in tubal pregnancy: prospective randomized trial using endoloop versus electrocautery
    Lim YH, Ng SP, Ng PH, Tan AE, Jamil MA
    J Obstet Gynaecol Res, 2007 Dec;33(6):855-62.
    PMID: 18001454
    Ectopic pregnancy is conventionally managed by laparoscopic salpingectomy. Electrocautery has been used widely to secure hemostasis during salpingectomy. However, this method is associated with a risk of thermal injury to the visceral organs. Endoloop, a pre-tied suture used in laparoscopic surgery may be an alternative treatment tool and its potential use in the management of ectopic pregnancy is explored here. Our study aims to compare the effectiveness of the endoloop technique to electrocautery during laparoscopic salpingectomy for tubal pregnancy.
  12. Effects of keratinocyte growth factor on skin epithelial differentiation of human amnion epithelial cells
    Fatimah SS, Tan GC, Chua K, Tan AE, Nur Azurah AG, Hayati AR
    Burns, 2013 Aug;39(5):905-15.
    PMID: 23273814 DOI: 10.1016/j.burns.2012.10.019
    The aim of the present study was to determine the effects of KGF on the differentiation of cultured human amnion epithelial cells (HAECs) towards skin keratinocyte. HAECs at passage 1 were cultured in medium HAM's F12: Dulbecco's Modified Eagles Medium (1:1) supplemented with different concentrations of KGF (0, 5, 10, 20, 30 and 50 ng/ml KGF). Dose-response of KGF on HAECs was determined by morphological assessment; growth kinetic evaluation; immunocytochemical analysis; stemness and epithelial gene expression quantification with two step real time RT-PCR. KGF promotes the proliferation of HAECs with maximal effect observed at 10 ng/ml KGF. However, KGF decreased the stemness genes expression: Oct-3/4, Sox-2, Nanog3, Rex-1, FGF-4, FZD-9 and BST-1. KGF also down-regulates epithelial genes expression: CK3, CK18, CK19, Integrin-β1, p63 and involucrin in cultured HAECs. No significant difference on the gene expression was detected for each Nestin, ABCG-2, CK1 and CK14 in KGF-treated HAECs. Immunocytochemical analysis for both control and KGF-treated HAECs demonstrated positive staining against CK14 and CK18 but negative staining against involucrin. The results suggested that KGF stimulates an early differentiation of HAECs towards epidermal cells. Differentiation of KGF-treated HAECs to corneal lineage is unfavourable. Therefore, further studies are needed to elucidate the roles of KGF in the differentiation of HAECs towards skin keratinocytes.
  13. Potential of human decidua stem cells for angiogenesis and neurogenesis
    Hayati AR, Nur Fariha MM, Tan GC, Tan AE, Chua K
    Arch Med Res, 2011 May;42(4):291-300.
    PMID: 21820607 DOI: 10.1016/j.arcmed.2011.06.005
    Placenta as a fetomaternal organ is a potential source of fetal as well as maternal stem cells. This present study describes novel properties of the cells isolated from the maternal part of term placenta membrane, the decidua basalis.
  14. Stemness and angiogenic gene expression changes of serial-passage human amnion mesenchymal cells
    Fatimah SS, Tan GC, Chua K, Fariha MM, Tan AE, Hayati AR
    Microvasc Res, 2013 Mar;86:21-9.
    PMID: 23261754 DOI: 10.1016/j.mvr.2012.12.004
    Particular attention has been directed towards human amnion mesenchymal stem cells (HAMCs) due to their accessibility, availability and immunomodulatory properties. Therefore, the aim of the present study was to determine the temporal changes of stemness and angiogenic gene expressions of serial-passage HAMCs.
  15. Fulltext The role of serum insulin-like growth factor I (IGF-I) in neonatal outcome
    Hayati AR, Cheah FC, Yong JF, Tan AE, Norizah WM
    J Clin Pathol, 2004 Dec;57(12):1299-301.
    PMID: 15563671
    AIMS: To determine the role of serum insulin-like growth factor I (IGF-I) in predicting the occurrence of septal hypertrophic cardiomyopathy in infants of mothers with diabetes.
    METHODS/MATERIALS: In this prospective study, 100 pregnant women (50 with diabetes and 50 controls), matched for age and race, were studied. One intrapartum blood sample was taken at 28 weeks of gestation from both groups of mothers and another sample at delivery. All samples were analysed for maternal IGF-I by an enzyme linked immunosorbent assay method. A chest radiograph and an electrocardiogram were performed on the babies of the mothers with diabetes within the first 24 hours of life. An echocardiogram was performed in the first 3 days of life to look for septal hypertrophy and to measure the myocardial thickness.
    RESULTS: In the six cases of neonatal septal hypertrophic cardiomyopathy, all the mothers had greatly raised IGF-I concentrations of more than 400 ng/ml at the time of delivery compared with a mean (SD) of 302 (25) ng/ml in control mothers.
    CONCLUSIONS: In the present study a crude analysis revealed that increased IGF-I concentrations correlate with neonatal septal hypertrophic cardiomyopathy.
    Study site: Obstetric and gynaecology clinic, Pusat Perubatan Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia
  16. Umbilical artery resistance index in diabetic pregnancies: the associations with fetal outcome and neonatal septal hypertrophic cardiomyopathy
    Tan AE, Norizah WM, Rahman HA, Aziz BA, Cheah FC
    J Obstet Gynaecol Res, 2005 Aug;31(4):296-301.
    PMID: 16018775 DOI: 10.1111/j.1447-0756.2005.00291.x
    Aim: To determine the incidence of an abnormal umbilical artery resistance index (UARI) in diabetic pregnancies and the relation to fetal outcome and the development of neonatal septal hypertrophic cardiomyopathy.

    Methods: A case-control study with subjects comprising 50 randomly selected diabetic mothers and a matched control group of 50 non-diabetic pregnancies. Doppler studies of the UARI were carried out at least once per week, beginning from 36 weeks' gestation for both groups. Within 48 h post delivery, echocardiograms were carried out on the newborn infants to identify those with hypertrophic cardiomyopathy, particularly asymmetrical septal hypertrophy.

    Results: The numbers of patients with abnormal UARI were similar in both the diabetic and control groups. A higher proportion of operative deliveries for intrapartum fetal distress was seen in patients with an abnormal UARI in the diabetic group. However, the groups did not differ in the numbers of infants who were small for gestational age, who had low Apgar scores or umbilical artery acidosis, and who required admission to the special care nursery. Six infants of diabetic mothers (12%) had septal hypertrophy, but none of these were associated with abnormal antenatal UARI.

    Conclusion: Diabetic pregnancy is not associated with a significantly higher incidence of abnormal UARI on Doppler study than non-diabetic pregnancy. UARI is not a useful single indicator by which to predict subsequent fetal outcome or the development of neonatal septal hypertrophic cardiomyopathy in diabetic pregnancies.
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