Urethral acellular matrix graft for repairing urethral defect in rabbits / 南方医科大学学报

<p><b>OBJECTIVE</b>To assess the biocompatibility of a urethral acellular matrix graft (UAMG) and evaluate its effect in repairing urethral defect in rabbit models.</p><p><b>METHODS</b>The UAMG was prepared and its structural features were observed using optical and electron microscopy. In vitro cultured rabbit bladder smooth muscle cells were seeded on UAMG and the cell proliferation was observed. The cytotoxicity of the aqueous extract of the UAMG against the cells was evaluated by MTT assay, and its biocompatibility was assessed by implanting the grafts subcutaneously on the back of the rabbits. In 24 male rabbits, a 2-cm urethral defect was induced and repaired with UAMG (experimental group, n=12) or left untreated (control group, n=12). In both groups, the rabbits were sacrificed 2, 4, 8 and 12 weeks after the operation for histological and immunohistochemical examination of the tissue regeneration.</p><p><b>RESULTS</b>The UAMG had a reticular fibrous structure without cell residues. The bladder smooth muscle cells showed normal proliferation on UAMG with normal cell morphology. The rabbits receiving the implants showed no abnormal response, and the UAMGs gradually degraded in vivo with grade 0 or 1 cytotoxcity showing satisfactory cytocompatibility. In the experimental group, new urethral tissues that were histologically compatible with normal urethral tissues were regenerated in the defect area 12 weeks after UAMG implantation.</p><p><b>CONCLUSION</b>As a tissue engineered scaffold material for urethral reconstruction, the UAMG possesses good biocompatibility and can induce the regeneration of urethral epithelial cells and smooth muscle cells.</p>

Laparoscopic treatment of a calcium fluorophosphate stone within a seminal vesicle cyst / 亚洲男科学杂志(英文版)

Stones in the seminal vesicles are extremely rare. We present a 62-year-old patient with a stone within a seminal vesicle cyst, who was cured by laparoscopic treatment. The operative time was 80 min, and the estimated blood loss was 90 mL. Scanning electron microscope examination of the stone showed a compact crystal image externally and sparse spherical crystal structure in kernel. Composition of the stone was calcium fluorophosphate on X-ray diffractometer. The follow-up time was 15 months with no recurrence of cyst or stone. To our knowledge, this case is the first to describe laparoscopic removal of a stone within a seminal vesicle cyst, and the first to describe calcium fluorophosphate as the composition of seminal vesicle stones.

Expression of kinase insert domain-containing receptor in prostate adenocarcinoma / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the correlation between the expression of the kinase insert domain-containing receptor (KDR) and the histological grade of prostate adenocarcinoma.</p><p><b>METHODS</b>Forty-eight samples of prostate adenocarcinoma tissues and 20 samples of benign prostatic hypertrophy (BPH) tissues were studied by LsAB immunohistochemical staining.</p><p><b>RESULTS</b>The positive expression rate of KDR was 73% in prostate adenocarcinoma and 30% in BPH. The expression of KDR was stronger in prostate adenocarcinoma, and there was no relationship between staining intensity and the histological grade of carcinoma.</p><p><b>CONCLUSION</b>KDR, expressed more highly in prostate adenocarcinoma, promises to be a new target in the treatment of prostate adenocarcinoma.</p>

Effects of experimental varicocele on the testosterone level in the serum and testis of rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the effects of experimental varicocele (VC) on the serum testosterone (T) and intratesticular testosterone in adolescent rats.</p><p><b>METHODS</b>A VC rat model was established by partial ligation of the left kidney vein in 20 SD rats, and another 20 were included in a sham operation group as controls. At 4 and 8 weeks, the concentrations of the serum T and intratesticular T were measured by radioimmunoassay (RIA). The testis tissues were homogenized and the extract liquid taken for RIA.</p><p><b>RESULTS</b>Compared with the controls, the level of serum T declined at 4 and 8 weeks in the VC group, but not significantly (P > 0.05), so was that of bilateral intratesticular T at 4 weeks, and with statistical significance at 8 (P < 0.01).</p><p><b>CONCLUSION</b>Within 8 weeks, experimental VC could reduce the level of bilateral intratesticular T, but not that of serum T. Varicocele could damage Leydig cells.</p>

Construction of tissue-engineered urothelial tissue in vitro and in vivo with bladder acellular matrix as scaffold / 中南大学学报(医学版)

OBJECTIVE@#To investigate the construction of urothelial structure by tissue engineering.@*METHODS@#Fresh bladder of New Zealand white rabbits were processed to prepare the bladder acellular matrix graft (BAMG) as the scaffold, which was evaluated by Masson's trichrome staining and the scanning electronic microscope. Bladder epithelia were obtained by enzymatic digestion and were proliferated in vitro. Growth of cells was observed under the inverted phase contrast microscope, and cells were identified by immunohistochemical method. The bladder epithelia were seeded on BAMG, and the epithelia/BAMG composites were observed by HE staining and the scanning electronic microscope. The composites fabricated in vitro were implanted into nude rats, and were retrieved in 4 and 8 weeks, which were observed by general observation, histological and immunohistochemical method.@*RESULTS@#White semi-transparent membrane appeared in the prepared BAMG, and a fibre mesh structure of the material without residual cells was observed under the scanning electronic microscope. Bladder epithelia cultured in vitro showed a paving stone structure. Immunohistochemical staining with cytokeratin was performed, and brown cellular plasma staining was observed as positive reaction. After the epithelia were seeded on BAMG in vitro for 7 days, the cells fully covered the surface of the framework, showing a single-layer cellular structure. After being implanted into nude rats for 4 weeks and 8 weeks, the epithelia seeded on the BAMG formed a multi-layer structure.@*CONCLUSION@#Urothelial structures can be constructed in vitro and in vivo by tissue engineering, which lays a technical foundation for further tissue engineered urinary tract reconstruction experiments.

Effect of KLF6 on prostate cancer cell line PC-3 by transgenic method / 中华男科学杂志

<p><b>OBJECTIVE</b>To observe the effect of KLF6 on prostate cancer cell line PC-3 by transgenic method.</p><p><b>METHODS</b>We obtained KLF6 cDNA by RT-PCR method from the liver cell, transfected plasmid pEGFP-C, recombinated with KLF6 into PC-3 cells, and used them as a transfection group and a control group. MTT, flow cytometer and immunocytochemical methods were used to observe the effect of anti-oncogene wild type KLF6 on prostate cancer cell line PC-3 by transgenic method for 48 hours.</p><p><b>RESULTS</b>After transfected into PC-3 cells, KLF6 enhanced growth suppression, (30.0 +/- 5.4)% in the transfection group and 0% in the control, P < 0.01, apoptosis, (24.3 +/- 2.3)% in the transfection group and (5.2 +/- 0.7)% in the control, P < 0.01, the down-regulation of the expression of cyclin D1, (25.3 +/- 3.7)% in the transfection group and (38.5 +/- 4.6)% in the control, P < 0.05 and Bcl-2, (18.7 +/- 3.2)% in the transfection group, and (41.8 +/- 5.9)% in the control, P < 0.01 in PC-3 cells. It also decreased the ratio of the cell phase G2/M, increased the ratio of G0/G1 from (58.6 +/- 7.3)% in the control to (80.0 +/- 9.8)% in the transfection group, P < 0.05.</p><p><b>CONCLUSION</b>PC-3 cells transfected with wild type KLF6 can enhance its growth suppression and apoptosis. It shows great potential for the gene therapy of androgen-independent carcinoma of the prostate.</p>

Risk factors for prostatic inflammation extent and infection in benign prostatic hyperplasia / 亚洲男科学杂志(英文版)

<p><b>AIM</b>To investigate the risk factors for prostatic inflammation extent and infection in patients with benign prostatic hyperplasia (BPH) so as to manage prostatic inflammation more efficiently.</p><p><b>METHODS</b>Sixty patients with BPH undergoing TURP between September 2005 and December 2005 in West China Hospital of Sichuan University were studied. Prostate fluid (PF) was collected for the measurement of secretory IgA (SIgA) and complement 3 (C3). Prostate tissue were collected for testing bacterial 16S rDNA by real-time PCR, examining SIgA in the tissue and examining the inflammation. The possible clinical and immune risk factors for prostatic inflammation or infection were analyzed by using the logistic regression method.</p><p><b>RESULTS</b>Abnormal white blood cell count in urinalysis, prostatic infection and a high concentration of C3 in PF are the risk factors for prostatic inflammation extent (P = 0.025, 0.034 and 0.035, respectively and odds ratio [OR] = 18.269, 8.284 and 1.508, respectively). Risk factors for prostatic infection include the C3 concentration and the concentration of SIgA in PF (P = 0.003 and 0.013, respectively, and OR=1.645 and 0.993, respectively).</p><p><b>CONCLUSION</b>The present study suggests that prostatic inflammation is associated with urinary tract infection, prostatic infection and the activated complement and that prostatic infection is associated with the activated complement and downregulated mucosal immunity in prostates of the patients with BPH. It is also suggested that individual immune regulation should be considered in the treatment of prostatic inflammation and infection of patients with BPH.</p>

Apoptosis in male germ cells induced by testicular denervation / 中华男科学杂志

<p><b>OBJECTIVE</b>To evaluate the role of spermatic nerves in the regulation of spermatogenesis.</p><p><b>METHODS</b>Fifty-four mature SD male rats (350-375 g) were randomized into a sham operation group (SO) and three experiment groups, and the latter underwent bilateral surgical removal of the superior spermatic nerve (SSN) or/and the inferior spermatic nerve (ISN). The animals were killed 1 month and 2 months after the operation. HE stain was used to observe spermatogenesis. Transmission electron microscopy and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) were employed to detect apoptosis.</p><p><b>RESULTS</b>Impaired spermatogenesis was observed 2 months after the operation, with only Sertoli cells and a few spermatogonia remaining in the regressed tubules in all the treatment groups. The abnormal tubules in the SSN, ISN and SSN + ISN denervated testes accounted for (13.25 +/- 2.03)%, (11.0 +/- 4.36)% and (34.17 +/- 3.78)% respectively. Chromosome condensation and fragmentation in the germ cells were observed under the electron transmission microscope in all the denervated testes. TUNEL showed the spermatogonia and Leydig cells to be apoptotic in all the denervated testes and the incidence of the apoptotic cells in the SSN + ISN denervated testes was significantly higher than in the SSN or ISN denervated ones.</p><p><b>CONCLUSION</b>Spermatic nerves play an important role in spermatogenesis.</p>

Expression of Bax and Bcl-2 gene in rat testis tissues induced by testicular denervation / 中华男科学杂志

<p><b>OBJECTIVE</b>To detect the changes of the expression of Bax and Bcl-2 gene in the denervated testis, and to explore the possible mechanisms underlying the apoptosis of germ cells induced by testicular denervation at the genetic translation level.</p><p><b>METHODS</b>Eighteen mature SD rats (350-375 g) were equally divided into 3 groups: a sham operation group( SO) , a superior spermatic nerve group (SSN) and an inferior spermatic nerve group (ISN) , and the latter two received bilateral surgical removal of the superior spermatic nerve and the inferior spermatic nerve, respectively. The animals were killed I month after the operation. ISH SP-method was used to detect the expression of Bax and Bcl-2 protein.</p><p><b>RESULTS</b>Significant up-regulation of Bax protein was detected in both the treatment groups 1 month after surgery( P <0. 05) , and the level of Bcl-2 protein remained unchanged.</p><p><b>CONCLUSION</b>Bax gene is involved in the apoptosis of germ cells induced by testicular denervation.</p>

Determination of expression of protamine-2 mRNA in different positions of the testis of patients with nonobstructive azoospermia / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the relationship between the expression of protamine-2 (P-2) mRNA and the results of sperm extraction in the corresponding testis tissues of patients with nonobstructive azoospermia.</p><p><b>METHODS</b>Based on pathological diagnosis, 38 cases of azoospermia at the mean age of 32.4 (ranging 24 - 42) years were divided into a nonobstructive (NOA) group and an obstructive (OA) group. Two specimens were taken from different positions of one testis, each divided into three portions for general pathological test, sperm separation and mRNA extraction, respectively. The expression of P-2 mRNA was determined by RT-PCR and image analysis assay.</p><p><b>RESULTS</b>Among the 38 cases, 27 were diagnosed as nonobstructive and 11 as obstructive azoospermia. No regularity was found as to the positions where sperm could or could not be successfully isolated. The expression of P-2 mRNA was 1.40 +/- 0.21 in the tissues where sperm was isolated and 0.51 +/- 0.23 (P < 0.05) in those where no sperm was isolated.</p><p><b>CONCLUSION</b>The expression of P-2 mRNA in the testicular tissues from the patient with nonobstructive azoospermia could reveal the results of sperm extraction in the corresponding tissues.</p>

Effects of heating on the epithelia of benign prostate hyperplasia in vitro / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the biological changes of the epithelia of benign prostate hyperplasia under heating at different temperatures and at different points of time after heating.</p><p><b>METHODS</b>Cell morphology, MTT, flow cytometer, and the immunocytochemical method for detecting heat shock protein 70 and prostate specific antigen were used to observe the effect of heating on the primarily cultured epithelia of benign prostate hyperplasia at 45 degrees C and 60 degrees C and at 1 hour and 12 hours after heating.</p><p><b>RESULTS</b>Heating at 45 degrees C for 15 min resulted in apoptosis, and at 60 degrees C, necrosis in most of the cells. The inhibiting effect of heating on the growth of cells was observed, more significant in the 60 degrees C group than in the 45 degrees C group. The cell phase arrest induced by heat, mainly G0/G1 arrest, was more significant in the 12 h group than in the 1 h group. Heating up-regulated the expressions of heat shock protein 70 and prostate specific antigen in cells.</p><p><b>CONCLUSION</b>Heating can induce apoptosis, necrosis and growth suppression of the epithelia of benign prostate hyperplasia. Its process and mechanism are correlated with the cell phase arrest and the up-regulation of the expressions of heat shock protein 70 and prostate specific antigen.</p>

Study on Toll-like receptors expression and cytokine production induced by bacillus Calmette-Guerin in human bladder cancer cell / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the effect of bacillus Calmette-Guerin (BCG) on Toll-like receptors (TLRs) expression and cytokine production in human bladder cancer cell line T24.</p><p><b>METHODS</b>Human transitional carcinoma cell line T24 was cultured in RPMI1640 medium with 10% FBS, BCG was added into the cell culture with various doses in bacteria-cell ratio. After T24 cells were stimulated by BCG for 1 hour, total RNA of cells was extracted. RT-PCR procedure was conducted with the primer of TLR-2 and TLR-4, and the products were analyzed with agarose gels electrophoresis. Then the expression of TLR-2 and TLR-4 in T24 cells was assessed by the analyzing system of gel imaging. After the stimulation culture for 12 hours, the supernatant of cells was collected. The levels of IL-4 and IL-12 in each sample were assayed by ELISA method.</p><p><b>RESULTS</b>The expression level of either TLR-2 or TLR-4 was increased by the stimulation of BCG in a dose-dependent manner, the effects reached the maximal level at the dose of BCG as 10 bacilli per cell. The production of IL-12 in T24 cells was also increased gradually by the stimulation of BCG in a dose-dependent manner, and the dose of BCG obtained maximal effect was 10 bacilli per cell, which is coincident with the results observed in the expression of TLRs. There was no difference showed on the production of IL-4 between T24 cells stimulated with BCG and control.</p><p><b>CONCLUSIONS</b>The stimulation of BCG not only up-regulated the expression of TLR-2 and TLR-4, but also increased the production of IL-12 in bladder cancer T24 cell line. The expression of TLRs and the production of cytokine in bladder cancer cells may be related to the BCG-induced immunol response to human bladder cancer.</p>

Clinical study on application of bailing capsule after renal transplantation / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe and assess the immunosuppressive effect of applying bailing capsule (BLC, a dry powder preparation of Cordyceps sinensis mycelia), after renal transplantation, its influence on other systems of organism, and to explore the possible therapeutic mechanism.</p><p><b>METHODS</b>One hundred and twenty-one recipients of renal homo-allograft were randomly divided into two groups. The 64 cases in Group A was treated with cyclosporin A (Cs A) + prednisone (pred) + azathioprine (Aza), the 57 in Group B treated with Cs A + pred + BLC. They were followed-up for 1-2 year by checking up blood routine, urine routine, liver and renal function, blood electrolytes, glucose and lipids, and uric acid for 2 times every week in the first month after transplantation, followed by proper re-examination of these items according to various condition.</p><p><b>RESULTS</b>There was no significant difference between the two groups in aspects of graft survival rate, occurrence of reject reaction, renal function recovery, blood electrolytes and blood glucose levels. However, as compared with Group A, in Group B, levels of urinary erythrocytes and leucocytes, blood alanine transaminase (ALT), aspartate aminotransferase (AST), total cholesterol, uric acid as well as the incidence of infection were significantly lower, and blood high density lipoprotein, serum total protein, albumin, RBC and WBC count were significantly higher.</p><p><b>CONCLUSION</b>BLC could effectively prevent the reject response after renal transplantation, protect renal and liver function, stimulate hemopoietic function, improve hypoproteinemia and hyperlipidemia, reduce the infection, etc., therefore, it is an ideal immunosuppressor after organ transplantation.</p>