Methylation and expression levels of microRNA-23b/-24-1/-27b, microRNA-30c-1/-30e, microRNA-301a and let-7g are dysregulated in clear cell renal cell carcinoma.

BACKGROUND: Renal cell carcinoma is the most common form of kidney cancer in adults. DNA methylation of regulatory sequences at the genomic level and interaction between microRNAs and the messenger RNAs of target genes at the posttranscriptional level contribute to the dynamic regulation of gene activity. Aberrations in these mechanisms can result in impaired functioning of cell signaling pathways, such as that observed in malignant tumors. We hypothesized that microRNA genes methylation may be associated with renal cancer in patients. METHODS AND RESULTS: We examined methylation levels of 22 microRNA genes in tumor and normal kidney tissue of 30 patients with TNM Stage III clear cell renal cell carcinoma using a pathway-specific real-time polymerase chain reaction array (EpiTect Methyl II PCR Arrays, Qiagen). MicroRNA expression analysis by quantitative polymerase chain reaction was also performed. Significant differences in methylation levels were found in two genes and in two clusters of microRNA genes. MicroRNA-23b/-24-1/-27b, microRNA -30c-1/-30e and let-7 g was hypermetylated in clear cell renal cell carcinoma tissue, microRNA -301a was hypomethylated in tumor compared with the adjacent normal tissues. Expression of microRNA-301a, microRNA-23b in the clear cell renal cell carcinoma tissues was significantly overexpressed when compared with the adjacent normal tissues and let-7 g was significantly downregulated in tumor. CONCLUSIONS: Our results may indicate the contribution of microRNA-301a, microRNA-23b and let-7 g in the pathogenesis of renal cancer, but further studies are needed to determine the functional significance of the detected changes.

Differential DNA Hydroxymethylation in Human Uterine Leiomyoma Cells Depending on the Phase of Menstrual Cycle and Presence of MED12 Gene Mutations.

Using immunofluorescence with specific antibodies, we analyzed DNA hydroxymethylation in uncultured cells from 25 human uterine leiomyomas considering the menstrual cycle phase during surgery and the presence of MED12 gene mutations. It was found that each tumor node had specific DNA hydroxymethylation level that did not depend on the presence of mutations in MED12 gene, but depended on the phase of menstrual cycle. The degree of DNA hydroxymethylation was significantly lower in cells of leiomyomas excised during the luteal phase compared to the follicular phase (p=0.0431). Hormonal status changing at various phases of menstrual cycle is a factor affecting DNA hydroxymethylation in leiomyoma cells.

[PELVIC FLOOR RECONSTRUCTION AFTER PELVIC EVISCERATION USING GRACILIS MUSCULOCUTANEOUS FLAP].

Evisceration of the pelvic organs (EPO) is a fairly uncommon surgical treatment that removes all organs from a patient's pelvic cavity. We use gracilis musculocutaneous flap to repair pelvic floor after EPO. Over the period from November 2013 to December 2014 we carried out EPO with reconstructive repair of the pelvic floor with gracilis musculocutaneous flap in 10 patients with locally advanced pelvic tumors. We describe the surgical procedure and surgical outcomes in these patients. Mean age of the patients was 55 years. Mean duration of EPO with the pelvic floor repair was 285 min., mean blood loss--595 mL and the average length of hospital stay--19 days. Gracilis musculocutaneous flap has a sufficient arterial supply and mobility for pelvic floor reconstruction. Necrosis of flap's distal edge occurred in one of the 10 clinical cases, while the remaining flaps were fully preserved. Complete healing of wounds with no signs of weakening of the pelvic floor muscles was observed in all cases. Pelvic floor reconstruction is an essential procedure in order to reduce complications associated with the evisceration of the pelvic organs. The Gracilis musculocutaneous flap is the logical alternative to repair pelvic floor defect. It does not contribute to complications like functional deficiency of the lower limbs, complications of stoma formation or weakening of the muscles of the anterior abdominal wall.

[MED12 gene mutations in women with uterine myoma].

Uterine leiomyoma (UL) is a benign and most common tumor that affects 20-45% of women of fertile age. In this study, we analyzed the MED12 second exon nucleotide sequence from 15 DNA samples extracted from LM of 15 subjects with uterine leiomyoma and 15 DNA samples extracted from peripheral blood leukocytes of the same female subjects. It was shown that somatic mutations in the MED12 gene occur in 73% of cases with deletions of varying sizes and missense mutations being most common at codon 44. Mutations in the MED12 gene could play an indirect role in leiomyoma progression by modifying the activity of other genes that encode proteins involved in growth and tumor progression.

[Combined therapy of trophic ulcers with protracted healing]. / Kompleksnaia terapiia dlitel'no ne zazhivaiushchikh troficheskikh iazv.

The results of treatment of 126 patients with trophic ulcers of the low extremities were analyzed. Trophic ulcers in 74 patients (58.7%) appeared due to postthrombophlebitic disease, in 25 patients (19.8%) due to primary varicose veins of lower limbs, in 15 (11.9%) patients due to obliterative diseases of peripheral vessels and in 12 (9.5%)--other diseases. The duration of the illness made up from 1 to 50 years. In 26 patients (group 1) low-intensity laser irradiation (LLI) was used together with combined conservative therapy; in 32 patients (group 2) hyperbaric oxygenation (HBO), in 21 (3 group)--combined application of LLI and HBO was used. The examination of regional macrohemodynamics and microcirculation in 39 patients showed decrease of rheographic index (RI) in the shin, increase of the arterioles tone and venous outflow disturbance. The use of LLI and HBO resulted in undoubtful increase of RI and improvement of microcirculatory indexes. As a result of combined application of LLI and HBO microflora of the ulcers lowered from 10 and 10 per 1 gr in patients of group 1 and 2, respectively, in patients of group 3 there was no growth of pathogenic microflora. The investigation of humoral and cellular immunity after the application of LLI and HBO demonstrated strong immunocorrective action.

[The chronic form of the Budd-Chiari syndrome]. / Khronicheskaia forma sindroma Badda-Kiari.

A rare chronic course of Budd-Chiari syndrome associated with thrombosis of the portal vein was observed in a 30-year-old male patient suffering from postmyocarditic cardiosclerosis. At the age of 24 the patient had infectious allergic myocarditis, was hospitalized and rehospitalized for circulatory insufficiency. Upon 3 years since the disease onset the patient was admitted to a hematological department for progressive enlargement of the spleen. The diagnosis on discharge was idiopathic myelofibrosis with portal hypertension. The treatment included prednisolone, blood transfusions, myelosan. In 1987 the patient presented with enlarged liver and spleen, ascites, gastric and esophageal varicosis, augmenting hepatic insufficiency clinically evaluated as hepatic cirrhosis. Postmortem examination revealed macrofocal cardiosclerosis, splenomegaly, ascites, portal varicosis, enlarged nutmeg liver with smooth surface. Microscopically there was phlebosclerosis and phlebothrombosis varying in duration and involving predominantly medial branches of the hepatic and portal veins, liver fibrosis. The findings provided evidence for the final diagnosis of Budd-Chiari syndrome running an uncommon chronic course.