AIM: For studies done in Malaysia, to identity the sample sizes and heterogeneity present in the various studies which used p16 in evaluating lesions of the cervix. To evaluate if it would be possible for a single study to answer the various questions posed by the original authors. To highlight areas where the design features of future studies can be optimised.
MATERIALS AND METHODS: Various databases were searched using synonyms for p16 AND cervix AND Malaysia. These were assessed for broad conformity to a Diagnostic Test Accuracy format. Methodological and clinical heterogeneity indicators were extracted into standardised fields.
RESULTS: There were 5 studies eligible for inclusion. Each sought to study different aspects of the disease such as diagnostic grade stratification and pathogenesis. The study type broadly conformed to a Diagnostic Test Accuracy format. The study design used was either consecutive or non-consecutive. Sample size ranged from 75 to 201. Clinical heterogeneity was present in the choice of controls with some using normal and some using inflamed tissue. Methodological heterogeneity in applying the reference test, index test and different antibody clones were present.
CONCLUSION: There was both clinical and methodological heterogeneity making synthesis of studies difficult. It is possible to design a study which would be able to answer all the questions posed by the original authors with internal validity while at the same time increasing sample size.
CASE REPORT: A 58-year-old female presented with a 3-cm painless right axillary mass. Extensive radiological investigations that include mammography, ultrasonography of the breasts and positron emission tomography (PET) scan failed to conclude the primary site of the tumour. Histological examination of the lymph node revealed loosely cohesive sheets of poorly differentiated malignant cells, without discernible glandular or squamous differentiation. Immunohistochemically, the malignant cells exhibited diffuse immunoreactivity toward pan-cytokeratin and CK7, while leukocyte common antigen, S100 and CK20 were negative. A second panel of immunomarkers was carried out. The malignant cells expressed breast-specific markers (GATA-3, GCDFP-15 and mammaglobin), and were negative for ER, PR and TTF-1 immunohistochemistry. A diagnosis of OBC was rendered.
DISCUSSION: Breast primary must always be considered in the differential diagnosis in patients with sole presentation of axillary lymphadenopathy. The breast-specific immunomarkers play a pivotal role in the diagnosis of ER, PR-negative occult breast cancer.
MATERIALS AND METHODS: The miR-302 cluster was amplified by polymerase chain reaction technique from human genomic DNA and was ligated into pTRIPz, an inducible lentiviral vector.
RESULTS: MRC5 fibroblasts and HEK293 (human embryonic kidney) cells were infected with pTRIPz-302 cluster lentivirus and the family of 302 miRNAs were strongly expressed in HEK293 cells but lowly expressed in MRC5 fibroblasts. When cultured in hESC conditions, MRC5 cells expressed only low levels of DNMT3B, Nanog, Oct4 and Lin28 and failed to show stem cell induction. The red fluorescent expression seen in the majority of MRC5 cells, indicated that the rate of infection by lentivirus was efficient.
DISCUSSION: The efficiency of reprogramming may be improved perhaps by either using a different cell type or a high expression vector with a different type of promoter.