[Expression of COX10 in human non-obstructive azoospermia testes].

OBJECTIVE: To evaluate the expression of COX10 mRNA in the testes of non-obstructive azoospermia patients and normal men. METHODS: A cDNA microarray containing COX10 and some other genes as RBM and EIF1AY was used to identify the differential gene expression profiles in the normal and azoospermic testes. The cDNA probes were prepared by labeling mRNA from azoospermic and normal testis tissues with Cy5-dUTP and Cy3-dUTP respectively through reverse transcription. The mixed cDNA probes were then hybridized with cDNA microarray. Later the fluorescent signals were scanned and the values of Cy5-dUTP and Cy3-dUTP on each spot were calculated and analyzed. After that an ISH was employed to detect the expression of COX10 mRNA in 10 fertile and 39 non-obstructive azoospermic testes, and the expression levels were compared to evaluate the significance. RESULTS: We obtained 128 differentially expressed genes that might be related with azoospermia, among which 56 were up-regulated and 72 down-regulated, with the expression of COX10 significantly decreased. In situ hybridization confirmed that the mRNA expression of COX10 was stronger in the spermatogenic cells of the normal fertile than the azoospermic testes. CONCLUSION: COX10 may play a certain role in the development and progression of azoospermia. The technique of cDNA microarray can be applied to further studies of screening non-obstructive azoospermia associated genes.

[Expression of cell cycle molecules in human azoospermic testes].

AIM: To evaluate the expression and significance of cell cycle molecules in human normal and azoospermia testes. METHODS: A cDNA microarray containing cDNA of some cell cycle molecules was used to identify the differential gene expression profiles between normal and azoospermic testes. cDNA probes were prepared by labeling mRNA from normal and testis tissues with Cy5-dUTP and Cy3-dUTP, respectively, through reverse transcription. Then the mixed cDNA probes were hybridized with cDNA microarray. The fluorescent signals were scanned, and the values of Cy5-dUTP and Cy3-dUTP on each spot were analyzed and calculated. After that an ISH was employed to detect the expression of CDC10 mRNA in ten fertility and thirty-nine azoospermic testes, whose expression level was compared to evaluate the significance. RESULTS: The genes which were differentially expressed in azoospermic testes were found, among which the expression of CDC7L1 and CDC10 was up-regulated but the expression of CDK9, CDC20 and CLK3 was down- regulated. The mRNA expression of CDC10 was confirmed to be stronger in spermatogenic cells of normal fertility compared with that of azoospermic testes by in situ hybridization. CONCLUSION: The cell cycle molecules such as CDC10, CDC7L1, CDK9, CDC20 and CLK3 may play a role in the development and progression of azoospermia.

[Expressions of cadherin molecules CDH18 and PCDH17 in human azoospermic testes].

OBJECTIVE: To investigate the expressions of cadherin molecules CDH18 and PCDH17 in normal and azoospermic human testes and their significance. METHODS: We studied the routine pathological slices of normal and non-obstructive azoospermic human testis tissues for changes in the tight junction of Sertoli-germ cells, and identified the differential gene expression profiles of the normal and azoospermic testis tissues using cDNA microarrays containing multiple cadherin molecules. The results were confirmed by Western blot. RESULTS: Abnormal tight junction of the Sertoli-germ cells was observed in 37.5% of the azoospermic testis samples, and obvious changes were seen in the expressions of some cadherin molecules, with down-regulation of CDH18 and PCDH17. CONCLUSION: Cadherin molecules such as CDH18 and PCDH17 may play a certain role in the development and progression of azoospermia, which might be related with the abnormal tight junction of the Sertoli-germ cells.

[A case report of simultaneous liver, pancreas-duodenum, and kidney transplantation in a patient with post-hepatitic cirrhosis combined with uremia and insulin-dependent diabetes related to chronic pancreatitis].

OBJECTIVE: To study the effect of triple organ transplantation (liver, kidney, and pancreas) in patient of end-stage liver disease with renal failure and diabetes, and to explore the optimal surgical procedure. METHODS: Simultaneous piggyback orthotopic heterotopic liver, pancreas-duodenum, and kidney transplantation was performed on a 43-year-old male patient with exocrine pancreatic insufficiency and insulin-dependent diabetes related to chronic pancreatitis (CP) who developed hepatic and renal failure. The pancreatic exocrine secretions were drained enterically to the jejunum. Prednisone, tacrolimus, mycophenolate mofetil, and ATG were used as immunosuppression therapy. RESULTS: Good liver and pancreas allograft function recovery was achieved within 7 days after the operation. And the recovery of renal allograft function was delayed. The renal allograft was removed because of break-down of renal blood flow 16 days after the transplantation. A new renal transplantation was performed at the same position. The second kidney graft recovered its normal function 3 days later. Up to the writing of this paper no acute rejection of organs and such complications as pancreatitis, thrombosis, and localized infection occurred. The patient became insulin independent with normal liver and renal function. CONCLUSION: Simultaneous piggyback orthotopic heterotopic liver, pancreas-duodenum, and kidney transplantation can be a good method for the patients with exocrine pancreatic insufficiency and insulin-dependent diabetes combined with hepatic and renal failure.

[Antisense oligonucleotide targeting survivin induces apoptosis of renal clear-cell carcinoma cells and enhances their sensitivity to epirubicin in vitro].

OBJECTIVE: To investigate the effect of antisense oligonucleotide (ASODN) targeting survivin on the apoptosis and proliferation of renal cancer cell line 786-O and enhancement of its sensitivity to epirubicin. METHODS: ASODN targeting survivin was designed and constructed. Cultured cells were divided into 6 groups: control group, liposome group, sense oligonucleotide (SODN) group, 600 nmol/L ASODN group, and 600 nmol/L ASODN combined with epirubicin group. After transfected for 24 h, cultured cells were harvested to carry on the next tests. Cell morphological changes were examined by transmission electron microscopy. Survivin protein was detected by immunohistochemical method. Apoptosis index (AI) and proliferation index (PI) were examined by flow cytometry. RESULTS: Morphological abnormalities of cells were observed in ASODN transfected groups. Expression of survivin in ASODN groups were significantly decreased compared with that in the control group, liposomes group and SODN group. AI of ASODN groups was significantly higher than that in other groups. PI of ASODN groups was significantly lower than that in other groups. The PI of ASODN combined with epirubicin group was (35.7 +/- 1.67)%, but (9.3 +/- 0.34)% or (8.5 +/- 0.21)% in liposomes group or SODN group that had combined with epirubicin. The ASODN group achieved the strongest effects to enhance apoptosis in comparison with control group (P < 0.05), while SODN did not cause statistically significant change (P > 0.05). CONCLUSION: The expression of survivin protein in the renal clear cell carcinoma cell line 786-O is downregulated by survivin ASODN. ASODN targeting survivin induces apoptosis and inhibits proliferation of 786-O cells. Inhibition of survivin enhances sensitivity of 786-O to epirubicin.

[Clinical investigation of renal angiomyolipoma].

OBJECTIVE: To study the diagnosis and management of renal angiomyolipoma (RAML), and to identify risk factors affecting spontaneous angiomyolipoma rupture. METHODS: The data of 68 patients with RAML from 1989 to 2002 were retrospectively reviewed. These patients were divided in two groups on the basis of tumor size, 35 patients in group A ( 4 cm). RESULTS: Seven patients were identified by image-guided percutaneous biopsy, and no major complications was noted. Sixteen patients with RAML were examined with angiography and 9 of 16 patients had got spontaneous rupture. 41.2% of patients were symptomatic, 4 cases (11.4%) in group A and 24 (72.7%) cases in group B (P < 0.01). There were significant differences in mean tumor size (11.6 cm +/- 5.1 cm vs 5.3 cm +/- 2.9 cm, P < 0.01) and mean aneurysm size (13.6 mm +/- 5.8 mm vs 2.6 mm +/- 3.0 mm, P < 0.01) between 9 cases of the ruptured tumor and 59 cases of unruptured tumor, 9 cases of the ruptured and 7 cases of unruptured tumor with angiography, respectively. Treatment consisted of conservative observation in 10 patients (no radiographic changes during the follow-up of 2 - 7 years); partial nephrectomy in 14 patients, tumor enucleation in 30 patients, total nephrectomy in 14 and posterior laparoscopic nephrectomy in 3 (no recurrence and complication correlation to operation during the follow-up of 2 - 144 months). CONCLUSIONS: It is an important role that percutaneous biopsy guided by ultrasonography or computerized tomography performs in managing suspicious and/or indeterminate RAML. A higher probability of rupture is related to tumor and/or aneurysms size. Nephron-sparing surgery is the first choice for surgical treatment of RAML.