Expression of miR-20a: A serum biomarker in the diagnostic approach for hepatocellular carcinoma.

OBJECTIVE: To determine the micro-ribonucleic acid-20a expression in hepatocellular carcinoma patients and its correlation with patients' demographic data, clinico-pathological and staging characteristics. METHODS: The case-control study was conducted from January to December 2014 at Isra University, Hyderabad, and comprised samples of hepatocellular carcinoma patients collected from two hospitals in Karachi and one in Hyderabad. Patients infected with chronic hepatitis B and C infections formed group-I and group-II and were compared with healthy controls in group-III. SPSS 21 was used for data analysis. RESULTS: There were 225 subjects divided into three equal groups of 75(33.3%) each. Among the controls, 57(76.0%) showed up-regulation, whereas 66(88.0%) in group-I and 50(66.7%) in group-II showed decreased expression. Micro-ribonucleic acid-20a showed significant down-regulation in group-I compared to group-II (p<0.001). Significant correlation of the down-regulation was found with male gender, age below 50 years and alphafetoprotein level below 20ng/ml (p<0.05). CONCLUSIONS: Micro-ribonucleic acid-20a expression was found to be decreased in hepatocellular carcinoma patients with significant correlation with gender, age and alpha-fetoprotein level..

Expression of miR-100 as a novel ancillary non-invasive biomarker for early detection of bladder carcinoma.

OBJECTIVE: To determine the expression of microribonucleic acid-100 in bladder cancer patients and to evaluate its association with clinicopathological parameters. METHODS: This cross-sectional case-control study was conducted at Jinnah Postgraduate Medical Centre Karachi and Civil Hospital Karachi from January to December 2014. Serum samples from patients having bladder carcinoma were collected, and divided into three equal groups. All the study participants were briefed about the study objectives, its outcomes, sample collection technique and their written consent was obtained. Serum samples of bladder carcinoma and healthy control subjects were divided into four equal groups. Group I included patients with papillary urothelial carcinoma of low malignant potential, group II had patients with papillary urothelial carcinoma low grade, and group III had patients with papillary urothelial carcinoma high grade. Group IV comprised healthy controls. From the serum samples, total ribonucleic acid were extracted, followed by micro ribonucleic acid extraction. The complementary deoxyribonucleic acid was synthesised and samples were further processed for real time polymerase chain reaction using the specifically designed primers of micro ribonucleic acid-100.Statistical analysis was done by SPSS 21. RESULTS: There were 240 subjects with 60(25%) in each of the four groups. Micro ribonucleic acid-100expression was decreased in all the three patient groups compared to the control group. A significant correlation was found between the expression of micro ribonucleic acid-100 with microscopic haematuria, cytology, cystoscopy and staging (p<0.05 each). No correlation was found between the expression and age, gender, gross haematuria and smoking (p>0.05 each). CONCLUSIONS: Micro ribonucleic acid-100was down-regulated in urothelial cancer patients with significant correlation with microscopic haematuria, cytology, cystoscopy and staging.

Role Of Alteration Of Ck5\6 Profile In Dysplastic Progression Of Oral Mucosa In Tobacco Users.

BACKGROUND: To determine staining expression of CK5\6 in healthy oral mucosa and various grades of oral dysplasia and to find out possible association of CK5\6 expression in dysplastic transformation of clinically normal oral mucosa to various grades of oral dysplasia. METHODS: This cross-sectional descriptive study was done at Al-Tibri Medical College and Hospital and Dow University of Health Sciences, Karachi from March 2018 to November 2018. It included 120 diagnosed paraffin embedded tissue samples of normal oral mucosa and various grades of oral epithelial dysplastic lesions. Patient's data was reviewed for age, gender and tobacco habits. For immunohistochemistry CK5\6 staining was performed on all the samples. Immunohistochemical evaluation was done by observing the staining expression of CK5\6 on various oral dysplastic samples on the basis of staining intensity. The compiled data was statistically analyzed by using Chi-square. p-values of <0.05 were considered to be significant. RESULTS: All of the 60/60 (100%) oral dysplastic cases were moderately to strongly positive for CK5\6. Gradual increase in staining intensity for CK5\6 was observed with increasing grades of dysplasia. We found highly significant association of CK5\6 immunopositivity in transformation of normal mucosa to various grades oral dysplastic lesions. CONCLUSIONS: CK5\6 can be used as reliable adjuvant marker for the early dysplastic transformation of oral mucosain tobacco users, before it progresses to oral squamous cell carcinoma (OSCC).

Flow Cytometric Analysis of ZAP-70 Protein Expression for B-Cell Chronic Lymphocytic Leukemia Prognostication: Usefulness and Limitations.

OBJECTIVES: The heterogenous clinical course in B-cell chronic lymphocytic leukemia (B-CLL) can be linked to several genetic and phenotypic characteristics of malignant B-cells. Prognostic analysis in B-CLL is routinely carried out to assist patient management; particularly to predict the time to initiate treatment. Increased ZAP-70 expression is a surrogate marker for unmutated immunoglobulin genes and inferior clinical outcomes which can be quantified to predict future outcomes in B-CLL patients. The study determined the ZAP-70 expression pattern using Z-index in Pakistani patients with B-CLL. METHODS: A retrospective analysis of B-CLL cases diagnosed and confirmed on flow cytometry at Aga Khan University Hospital for the last six years which had also undergone ZAP-70 analysis were included. In all these cases, ZAP-70 expression was quantified by measuring mean fluorescence intensities (MFIs) of normal B-cells, T-cells, and CLL-cells (CD19 and CD5 double-positive population). ZAP-70 expression was divided into high, low, and negative categories based on Z-index calculation. Mann-Whitney U test was utilized to determine the significance of ZAP-70 variations in different age groups and genders. P-value <0.05 was considered significant. RESULTS: A total of 120 patients of B-CLL had ZAP-70 analysis during the study period. The median age was 62 with an interquartile range of 35-87 and male to female ratio of 2:1. ZAP-70 expression was high in 18 (15%), low in 52 (43.3%) and negative in 50 (41.7%) cases. No significant difference in ZAP-70 expression with respect to the age or gender of the study population was identified using appropriate statistical calculations. CONCLUSIONS: This study showed only 15% of B-CLL cases showing high ZAP-70 expression, a surrogate biomarker for possible aggressive behavior which may necessitate therapeutic intervention and close surveillance.