Urinary exosomal microRNA-96-5p and microRNA-183-5p expression as potential biomarkers of bladder cancer.

Because of low sensitivity and specificity of the currently available urine biomarkers of bladder cancer (BC) detection and painful cystoscopy procedure. Our study aimed to evaluate expression of urinary exosomal miR-96-5p and miR-183-5p as probable non-invasive and accurate biomarkers for the diagnosis and follow up of BC. Using quantitative real-time polymerase chain reaction; expression of exosomal microRNA (miR)-96-5p and miR- 183-5p in the urine samples of 51 patients with BC, 21 patients with benign urinary bladder lesions and in 24 normal individuals as control group was done. Our study results showed higher expressions of both miR-96-5p and miR-183-5p in urine of BC patients in comparison with control group (P < 0.001 for each). Receiver operating characteristic curve (ROC) analysis showed that each microRNA had good sensitivity and specificity to differentiate BC from non-BC patients miR-96-5p 80.4% and 91.8% and miR-183-5p 78.4% and 81.6% respectively compared to cytology (37.3% and 100%). In addition, it was obvious that the sensitivity of combined miR-96-5p and miR-183-5p for the diagnosis of BC reached 88.2%% and specificity reached 87.8%, which were higher than each one alone. We also found that expression of miR-96-5p and miR-183-5p with high grade, and pathological stage was significantly increased. After surgery, collected urine samples showed significantly lower expression of miR-96-5p-: P < 0.001; and miR-183-5p: P = 0.002. In conclusion, urine miR-96-5p and miR-183-5p are promising tumor biomarkers of BC diagnosis; particularly, when they combined with each other or with urinary cytology.

The validity of testicular aspirate cytology and DNA image-analysis of the aspirate in the assessment of infertile men.

OBJECTIVE: To assess the possibility of using cytological examination and DNA image-analysis of testicular fine-needle aspirates instead of open surgical biopsy in the investigation of infertile men, as testicular biopsy has long been used for investigating infertility but the interpretation of histological slides is usually subjective. PATIENTS AND METHODS: Thirty-three men (aged 22-36 years) were evaluated for infertility and underwent both open biopsy and fine-needle aspiration of their testes. Subsequently, the needle aspirates were assessed histopathologically and cytologically, and by DNA image cytometry. The percentages of haploid, diploid and tetraploid cells were determined for each patient. RESULTS: The cases were divided into four categories: (1) Complete spermatogenesis, with a DNA pattern of 1n > 2n > 4n; (2) Maturation arrest, with a DNA pattern of 2n > 4n with no haploid cells; (3) Sertoli cell-only syndrome, with a DNA pattern of only 2n, with no haploid or tetraploid cells; (4) Hypospermatogenesis, with a variable DNA pattern, i.e. mild with 1n > 2n, moderate with 2n > 1n > 4n, and marked where the DNA pattern was 2n > 4n > 1n. From the cytological and DNA image-analysis of the aspirate a diagnosis was possible that had a strong correlation with the histological diagnosis of the same case. From image analysis we could exclude interstitial cells, Sertoli cells and sperms on the static image, and differentiate between spermatozoa and spermatids based on morphological characteristics in the cytological smear. This technique can therefore be used to quantitatively determine the percentages of various cell types within the seminiferous tubules. By coupling image ploidy analysis and cytological examination of a cytological smear, spermatogenesis can be assessed accurately. CONCLUSION: Image cytometry could be used to exclude interstitial cells, Sertoli cells and sperms on the static image and so produce an accurate assessment of spermatogenesis. A combination of ploidy and cell morphology characteristics in cytological smears provides an accurate, reproducible and easily used alternative to open testicular biopsy.