Phenotypic transition of corpus cavernosum smooth muscle cells subjected to hypoxia.

Corpora cavernosum smooth muscle cells (CCSMCs) have been shown to play a critical role in the male erectile response and are involved in the pathogenesis of multiple causes of erectile dysfunction (ED). To investigate the underlying mechanisms, we studied the changes that CCSMCs undergo under hypoxic conditions in vitro. The identified and characterized CCSMCs were exposed to hypoxia for 48 h and its phenotypic changes were examined by light and electron microscopy, MTS assay and flow cytometry. The mRNA and protein levels of TGF-ß1 and type I/III collagen, as well as CCSMC phenotype marker proteins and their transcriptional factors, were assessed by qPCR, immunofluorescence analysis and western blotting. Our results showed that CCSMCs became hypertrophic with loss of myofilament bundles and formation of an extensive rough endoplasmic reticulum (RER) under hypoxic conditions, with inhibited cell proliferation and enhanced cell apoptosis. This was accompanied by the increased synthesis of TGF-ß1 and types I and III collagen. Moreover, smooth muscle cell phenotypic markers were also affected by hypoxic conditions, as indicated by the decrease in α-SMA, desmin and CNN1 expression and the increase in vimentin expression. These changes corresponded to changes in associated transcriptional factors, such as the increase in Elk-1 and KLF-4 expression and decrease in Myocd expression. In addition, a HIF-1α knockdown effectively reversed the hypoxia-induced CCSMC phenotype, whereas its overexpression induced the dedifferentiation phenotype. These results indicate that CCSMCs undergo a phenotypic transition under hypoxic conditions.

Androgen deprivation induces bladder histological abnormalities and dysfunction via TGF-ß in orchiectomized mature rats.

Symptomatic late-onset hypogonadism, one of the most common elder diseases, is defined as a syndrome associated with a deficiency in serum testosterone. Recent studies have indicated that androgen deficiency in men is also associated with lower urinary tract symptoms and bladder dysfunction. To determine the pathologic consequences of androgen deprivation in bladder histology and function, we addressed the underlying mechanism. Male rats were divided into 4 groups: emasculated rats (EMR), emasculated rats treated with testosterone, emasculated rats treated with anti- transforming growth factor-ß (TGF-ß) neutralizing antibody, and sham surgery rats. TGF-ß is a common profibrotic factor that mediates the pathologic process of fibrosis in multiple organs. Two months later, urodynamic evaluations were employed to determine the bladder function in vivo. And then rats were sacrificed, and the bladder tissues were collected. Histological studies were employed to determine the degree of bladder fibrosis. Real time PCR was used to evaluate the mRNA level of pro-collagen I, a fibrotic marker. We demonstrate here that androgen deficiency induces bladder fibrosis and decreases the bladder maximal volume and compliance. Androgen replacement treatment completely prevented the histological and functional abnormalities induced by androgen deficiency. Subsequently, we identified that androgen deprivation induced the induction of TGF-ß mRNA level. Importantly, treatment with anti-TGF-ß antibody abolished androgen deprivation-induced bladder fibrosis and dysfunction. Our study reveals an essential role of TGF-ß in the pathogenesis of androgen deprivation-induced bladder fibrosis and dysfunction and offers a potential target for prevention and treatment of bladder dysfunction associated with androgen deficiency.

Nkx3.1 and p27(KIP1) cooperate in proliferation inhibition and apoptosis induction in human androgen-independent prostate cancer cells.

Prostate cancer (PC), which responds well to androgen ablation initially, invariably progresses to treatment resistance. The so-called androgen-independent PC is also a concern, since there is no effective therapy so far. Nkx3.1 is a putative prostate tumor suppressor that is expressed exclusively in the prostate under the regulation of androgen, and p27(KIP1) functions as a cell proliferation inhibitor and apoptosis trigger by disrupting the cyclin-dependent kinase (CDK)-cyclin complex. Lack of expressions of Nkx3.1 and/or p27(KIP1) have been detected in most advanced PC and is associated with poor clinical progression. Here, we show that endogenous expressions of both Nkx3.1 and p27(KIP1) are lost in the androgen-independent PC3 PC cells, while remaining intact in LNCaP PC cells, which contain functional androgen receptor (AR) and are hormone-responsive. Ectopic restoration of either Nkx3.1 or p27(KIP1) in PC3 cells results in reduced cell proliferation, and increased cell death. Both effects are synergistically enhanced when the two molecules are coexpressed. p27(KIP1) overexpression in PC3 results in increased cell population ceased at the G0/G1 phase, and this cell-cycle-arresting effect is significantly enhanced by the coexpression of Nkx3.1. Flow cytometry further revealed that Nkx3.1 and p27(KIP1) also cooperatively render more PC3 cells undergoing apoptosis. Consistently, the coexpression of Nkx3.1 and p27(KIP1) leads to the decreased expression of Bcl-2 oncogene and a concomitantly upregulated Bax expression. It also activates caspase 3 and leads to increased cleavage of PARP. Our findings thus reveal the crucial relevance of the combined antiproliferative and proapoptotic activities of Nkx3.1 and p27(KIP1) in androgen-independent PC cells, and further suggest that a combined, rather than single gene manipulation may be of clinical value for hormone-refractory PC.

[Simultaneous inhibition of both hTERT and androgen receptor gene expression in LNCaP cells using single shRNA vector].

OBJECTIVE: To explore the feasibility of inhibition of hTERT and androgen receptor (AR) gene expression simultaneously in LNCaP cells by single shRNA vector. METHODS: Templates DNA of both hTERT and AR siRNA were inserted into Pgenesil vector to construct a new vector Pgenesil-hTERT-AR-shRNA by RNAi-DNA vector technology. Pgenesil-HK-shRNA, Pgenesil-hTERT-shRNA, Pgenesil-AR-shRNA and Pgenesil-hTERT-AR-shRNA vectors were transfected into prostate cancer LNCaP cells respectively. The levels of AR mRNA, apoptosis and proliferation of each cell group were determined by FQ-PCR, Annexin V method and MTT. RESULTS: The level of hTERT mRNA of control group cells and cells transfected by Pgenesil-HK-shRNA, Pgenesil-hTERT-shRNA, Pgenesil-AR-shRNA and Pgenesil-hTERT-AR-shRNA was (1.51 +/- 0.08) x 10(8), (7.32 +/- 0.43) x 10(7), (2.94 +/- 0.15) x 10(6), (4.45 +/- 0.25) x 10(7) and (3.17 +/- 0.18) x 10(6) (copies/ml) respectively. The level of AR mRNA of control cell groups and cells transfected by Pgenesil-HK-shRNA, Pgenesil-hTERT-shRNA, Pgenesil-AR-shRNA and Pgenesil-hTERT-AR-shRNA was (1.92 +/- 0.11) x 10(5), (6.47 +/- 0.32) x 10(5), (3.70 +/- 0.24) x 10(4), (1.22 +/- 0.06) x 10(4) and (7.21 +/- 0.41) x 10(3) (copies/ml) respectively. These data indicate that the expression of hTERT or AR gene could be significantly inhibited by Pgenesil-hTERT-shRNA or Pgenesil-AR-shRNA while Pgenesil-hTERT-AR-shRNA could simultaneously inhibit both hTERT and AR gene expression. The apoptosis rate and the inhibition rate of cell growth of Pgenesil-hTERT-AR-shRNA group were significantly higher than those of Pgenesil-hTERT-shRNA group or Pgenesil-AR-shRNA group (P < 0.05). CONCLUSION: It is feasible to inhibit both hTERT and AR gene expression simultaneously by single shRNA vector. It will be a new research strategy of gene therapy for prostate cancer.

[Protective effect of MEK inhibitor (U0126) on donor testes from ischemia-reperfusion injury after orthotopic testicular transplantation in rats].

OBJECTIVE: To determine the effect of MEK inhibitor (U0126) on donor testes from ischemia-reperfusion injury after orthotopic testicular transplantation in rats. METHODS: The rats were divided into 7 groups, Group 1: normal control; Group 2: cold perfusion control; Group 3: sham operation control; Group 4: transplanted for 30 min; Group 5: transplanted for 1 week; Group 6: transplanted for 30 min with pretreatment of U0126; Group 7: transplanted for 1 week with pretreatment of U0126. The orthotopic testicular transplantation model was established with cuff. The levels of ERK1, ERK2, pERK1 and pERK2 of donor testes were evaluated; the change of histology and gonadal hormones were measured as well. RESULT: Group 1, 2 and 3 had no significant differences in all results (P>0.05). The levels of ERK1, ERK2, pERK1 and pERK2 in Group 4 were significantly increased compared with Group 1 (P<0.05), the levels of ERK1 and ERK2 in Group 6 were not different from those of Group 4 (P >0.05), but the levels of pERK1 and pERK2 in Group 6 were lower than those in Group 4 significantly(P <0.05), the histological changes in Group 6 were similar to Group 1 but milder than that in Group 4. The histological injury was more severe in Group 5 than that in Group 7, and the levels of gonadal hormones in Group 5 were lower than those in Group 7 (P <0.05) which remained at the normal levels. CONCLUSION: U0126 has a protective effect on the donor testes in a short period through inhibiting expression of pERK1/2 activated by testicular transplantation.

Oxidative stress stimulates testicular orphan receptor 4 through forkhead transcription factor forkhead box O3a.

Early studies reveal that testicular orphan nuclear receptor 4 (TR4) modulates signaling pathways that control various cell functions. However, how TR4 activity is regulated without the involvement of specific ligand(s) remains unclear. Here we identify a daf-16 family protein-binding element (DBE; 5'-TGTTTAC-3') in the TR4 promoter that can be recognized by the forkhead transcriptional factor FOXO3a, a key stress-responsive factor, through which TR4 gene expression is activated. The interaction between DBE and FOXO3a was confirmed using EMSA and chromatin immunoprecipitation assays. Activation of FOXO3a by oxidative stress and phosphatidylinositol 3-kinase inhibitor induced TR4 expression; in contrast, suppression of FOXO3a by small interfering RNA can reduce oxidative stress-induced TR4 expression. The biological consequence of the FOXO3a-induced TR4 by oxidative stress is to protect against stress-induced cell death in which cells with reduced FOXO3a are less resistant to oxidative stress, and addition of functional TR4 can increase stress resistance. These results suggest that this new identified oxidative stress-FOXO3a-TR4 pathway is a fundamentally important mechanism regulating stress resistance and cell survival.

Transgelin functions as a suppressor via inhibition of ARA54-enhanced androgen receptor transactivation and prostate cancer cell growth.

The androgen receptor (AR) requires coregulators for its optimal function. However, whether AR coregulators further need interacting protein(s) for their proper function remains unclear. Here we describe transgelin as the first ARA54-associated negative modulator for AR. Transgelin suppressed ARA54-enhanced AR function in ARA54-positive, but not in ARA54-negative, cells. Transgelin suppressed AR transactivation via interruption of ARA54 homodimerization and AR-ARA54 heterodimerization, resulting in the cytoplasmic retention of AR and ARA54. Stable transfection of transgelin in LNCaP cells suppressed AR-mediated cell growth and prostate-specific antigen expression, whereas this suppressive effect was abolished by the addition of ARA54-small interfering RNA. Results from tissue surveys showing decreased expression of transgelin in prostate cancer specimens further strengthened the suppressor role of transgelin. Our findings reveal the novel mechanisms of how transgelin functions as a suppressor to inhibit prostate cancer cell growth. They also demonstrate that AR coregulators, like ARA54, might have dual in vivo roles functioning as both a direct coactivator and as an indirect mediator in AR function. The finding that a protein can modulate AR function without direct interaction with AR might provide a new therapeutic approach, with fewer side effects, to battle prostate cancer by targeting AR indirectly.

Incidental prostate cancer in radical cystoprostatectomy specimens.

AIM: To investigate the rates of prostate cancer (PCa) in radical cystoprostatectomy (RCP) specimens for bladder cancer in mainland China. To determine the follow-up outcome of patients with two concurrent cancers and identify whether prostate-specific antigen (PSA) is a useful tool for the detection of PCa prior to surgery. METHODS: From January 2002 to January 2007, 264 male patients with bladder cancer underwent RCP at our center. All patients underwent digital rectal examination (DRE) and B ultrasound. Serum PSA levels were tested in 168 patients. None of the patients had any evidence of PCa before RCP. Entire prostates were embedded and sectioned at 5 mm intervals. RESULTS: Incidental PCa was observed in 37 of 264 (14.0%) RCP specimens. Of these, 12 (32.4%) were clinically significant according to an accepted definition. The PSA levels were not significantly different between patients with PCa and those without PCa, nor between patients with significant PCa and those with insignificant PCa. Thirty-four patients with incidental PCa were followed up. During a mean follow-up period of 26 months, two patients with PSA > 4 ng/mL underwent castration. None of the patients died of PCa. CONCLUSION: The incidence of PCa in RCP specimens in mainland China is lower than that in most developed countries. PSA cannot identify asymptomatic PCa prior to RCP. In line with published reports, incidental PCa does not impact the prognosis of bladder cancer patients undergoing RCP.

Effects of wortmannin on phosphorylation of PDK1, GSK3-beta, PTEN and expression of Skp2 mRNA after ischemia/reperfusion injury in the mouse kidney.

AIM: The aim of this study was to investigate the role of 3-phosphoinositide-dependent protein kinase-1 (PDK1), glycogen synthase kinase-3beta (GSK3-beta) and phosphatase and tensin homologue deleted on chromosome 10 (PTEN), which are the components of the phosphoinositide 3-kinase (PI3K)/Akt pathway, and expression of S-phase kinase-associated protein 2 (Skp2) messenger RNA (mRNA) in renal ischemia/reperfusion. MATERIALS AND METHODS: Three experimental groups, sham-operative mice, vehicle-delivered mice and wortmannin-treated ischemia/reperfusion injury (IRI) mice were designed to examine PDK1, GSK3-beta and PTEN phosphorylation status, as well as expression of Skp2 mRNA at 30 min, 90 min, 24 h and 48 h of reperfusion after ischemia treatment. Wortmannin or its vehicle was given intraperitoneally 4 h before surgery. Expression of Skp2 mRNA was examined by semiquantitative reverse-transcription PCR, and the components of the PI3K/Akt pathway were detected by western blotting in the IRI kidney. RESULTS: Phosphorylation of PDK1(Ser241), GSK3-beta(Ser9) and PTEN(Ser380) was increased after ischemia/reperfusion in the mouse kidney, and phosphorylation of PDK1 was reduced by wortmannin administration. The renal Skp2 mRNA increased after IRI in mouse, which could be inhibited by wortmannin. CONCLUSIONS: Our primary study suggests that the PI3K/Akt signaling pathway plays an important role in regulating the repair following renal IRI. The Skp2 mRNA increased in the IRI kidney and may be regulated by the PI3K/Akt pathway.

[Feasibility and safety evaluation of retrograde inserting of ureteric catheter via flexible cystoscope].

OBJECTIVE: To introduce a method of retrograde ureteric catheter placement via flexible cystoscope , and to evaluate the feasibility and safety of this method. METHODS: 112 patients, 62 males and 50 females undergoing retrograde ureteropyelography by 2 same physicians in cooperation were randomly divided into two equal groups with 31 males and 25 females each: one group via flexible cystoscope and the other group via rigid cystoscope. The catheterizing time, visual analogue scale (VAS) pain score, gross hematuria rate, and fever rate were compared between these 2 groups. RESULTS: Fifty-five patients underwent ureteric catheter placement successfully via flexible cystoscope (98%, 55/56), while 53 cases were technically successful by rigid cystoscope (95%, 53/56). The inserting time in women patients of the flexible cystoscopy group was (7.6 +/- 1.8) min, significantly shorter than that of the men [(8.0 +/- 1.8) min, P < 0.05]. The inserting time in women patients of the rigid cystoscopy group was (7.4 +/- 1.5) min, significantly shorter than that of the men [(8.2 +/- 1.2) min, P < 0.05]. However, there were not significant differences in the inserting times in both men and women between these 2 groups (both P > 0.05). The VAS pain scores in men and women of the flexible cystoscope group were 3. 5 and 2. 3 respectively, both significantly lower than those of the rigid cystoscopy group (7.2 and 3.3 respectively, both P < 0.05). The gross hematuria rate of the flexible cystoscope group was 8.6% (5/56), significantly lower than that of the rigid cystoscopy group (25.0%, 14/56, P < 0.05). Four patients had a fever after flexible cystoscopy while 6 cases did after rigid cystoscopy, however, without significant difference between these 2 groups (P > 0.05). CONCLUSIONS: Retrograde placement of ureteric catheter via flexible cystoscope is safe and reliable as rigid cystoscopy. Meanwhile, inserting ureteric catheter via flexible cystoscope causes the patients less pain and less chance of hematuria.

Expression of Nkx3.1 enhances 17beta-estradiol anti-tumor action in PC3 human prostate cancer cells.

AIM: To explore whether the anti-tumor action of 17beta-estradiol is enhanced by re-expression of the homeodomain transcription factor Nkx3.1 in PC3 human prostate cancer cells. METHODS: PC3 cells were stably transfected with pcDNA3.1-Nkx3.1-His vector, which carries a full-length cDNA of human Nkx3.1. The PC3 cells stably transfected with vector pcDNA3.1 were set as a control. The expression of Nkx3.1 protein in the cells was confirmed by Western blot analysis. The effect of Nkx3.1 on cell proliferation of PC3 cells was examined with MTT assay. The antiproliferative and apoptotic effects of 17beta-estradiol alone or in combination with Nkx3.1 were estimated on PC3 cells by using MTT growth tests and flow cytometric analyses. The expression of apoptosis-related proteins was analyzed using Western blotting. RESULTS: The plasmid carrying Nkx3.1 gene induced high expression of Nkx3.1 protein in PC3 cells. The re-expression of exogenous Nkx3.1 did not cause a significant reduction in cellular proliferation, whereas the expression of Nkx3.1 enhanced the 17beta-estradiol anti-proliferative effect in PC3 cells. Nkx3.1 expression promoted 17beta-estradiol-induced apoptosis of PC3 cells, as shown by analysis of Bcl-2, Bax, Caspase-3 and poly (ADP-ribose) polymerase expression. CONCLUSION: The present study demonstrates that re-expression of Nkx3.1 enhances 17beta-estradiol anti-tumor action in PC3 human prostate cancer cells. The in vitro study suggests that re-expression of Nkx3.1 is worthy of further consideration as an adjuvant treatment of androgen independent prostate cancer with estrogen anti-tumor therapies.

Expression of oestrogen receptor-alpha and oestrogen receptor-beta in prostate cancer.

BACKGROUND: Recent studies have suggested that estrogens are involved in normal and abnormal prostate growth, though their exact role is still controversial. Oestrogens exert inhibitory and stimulatory effects on prostate gland, but the expression of oestrogen receptor-alpha (ERalpha) and oestrogen receptor-beta (ERbeta) in malignant prostate tissue remains unresolved. We determined ERalpha and ERbeta in prostate cancer and investigated the relationship between expression of ER and pathological features of prostate carcinoma. METHODS: Thirty-two cases of prostate cancer, 12 cases of normal prostate tissue and 32 cases of benign prostate hyperplasia were analyzed for the expression of ERalpha and ERbeta using semiquantitative, reverse transcription polymerase chain reaction (RT-PCR) and the products sequenced. RESULTS: Comparisons of the normal, hyperplastic and tumour prostate tissues indicated an overexpression of ERalpha in tumour specimens (P < 0.01). However, the expression of ERbeta significantly reduced in tumour tissues compared with normal and hyperplastic specimens (P < 0.01), suggesting that severe pathological features of prostate cancer were associated with lower ERbeta expression. Spearman analysis showed negative correlation between ERbeta expression and tumour stage, grade (-0.67, -0.43, respectively, both P < 0.05), and a positive correlation between ERalpha expression and tumour stage, grade (0.51, 0.57, respectively, both P < 0.01). Our analysis also showed that hormone refractory, prostate cancer, compared with hormone dependent, prostate cancer, displayed a decreased expression of ERbeta (P < 0.01) and an increased expression of ERalpha. CONCLUSIONS: ERalpha and ERbeta may play important roles in the development of prostate cancer. The decrease in ERbeta expression is associated with higher Gleason grade tumours and prostate cancer with higher metastatic potential. The loss of ERbeta could be one of the key processes leading to uncontrolled growth of prostate epithelial cells.

[Hypoxia-inducible factor 1 and prostate cancer].

Hypoxia-inducible factor 1 (HIF-1) has a close relation with prostate cancer. It is involved not only in angiogenesis, cell proliferation/survival and glucose metabolism but also in p53, p21 and signal transduction pathway in prostate cancer. Further studies of HIF-1 may yield new approaches to the diagnosis and treatment of prostate cancer. We present a review of the structure and biological functions of HIF-1 and its relation with prostate cancer.

Effect of cocaine on germ cell apoptosis in rats at different ages.

AIM: To investigate the effect of cocaine on apoptosis and caspase-3 activity in germ cells in male rats at different ages. METHODS: Cocaine hydrochloride was given (15 mg/kg body weight s.c.) to male Sprague-Dawley rats of 3 weeks (n = 8), 6 weeks (n = 8) and 12 weeks (n = 8) of age, daily for 28 Days. The serum levels of follicle stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL), testosterone (T) and estrogen (E2) were assayed, and the DNA fragmentation of germ cells was determined by gel eletronphoresis. The cell cycle, apoptosis and caspase-3 activity of germ cells were tested by flow cytometry. RESULTS: After the 28-day cocaine treatment, testes weight of the 3-week-old rats, the testes and body weights of the 6-week-old rats were decreased significantly compared to those of their corresponding controls (P < 0.05). The serum level of T was decreased significantly in the 3-week-old and 6-week-old rats, and the serum level of PRL was also decreased significantly in 12-week-old rats compared to the controls (P < 0.05). In all the three cocaine-treated groups, the isolated DNA displayed a clear ladder pattern, especially in the 6-week old rats. The number of apoptosic germ cells increased significantly in 3- and 6-week-old rats treated with cocaine (P < 0.05). The caspase-3 activity in all three groups increased significantly compared to the controls (P < 0.05), especially in the 6-week-old rats. CONCLUSION: Cocaine exposure for 28 Days leads to significant damage to male gonad and apoptosis elevation in testes of rats of different ages, especially in those of 6 weeks of age. The increase in caspase-3 activity might be a key pathway related to the early stage of apoptosis as the mechanism of cocaine-induced germ cell loss.

[Establishment of testis transplantation model and study on mechanism of graft injury in rats].

OBJECTIVE: To establish the testis transplantation model in rats and to study the mechanism of graft injury. METHODS: The testis orthotopic transplantation model was established using three-cuff method. The animals were divided into 6 groups. Serum levels of testosterone (T), luteining hormone (LH) and follicle-stimulating hormone (FSH) were determined by radioimmunoassay (RIA). Morphology and ultrastructure were examined by light and electron microscopy. Expression of Glial cell line-derived neurotrophic factor (GDNF) mRNA was studied by reverse-transcription polymerase chain reaction (RT-PCR) technique. RESULT: On the 7th day postoperatively, the allotransplanted testes showed perivascular massive infiltration of lymphocytes and polymorphonuclear neutrophil (PMN) and reduced number of the sertoli cells under light microscopy. It also showed the broken blood-testis barrier, the atrophy of the sertoli cells and spermatogenic cells arranged in disorder under electron microscopy. The decline of serum T level and the increase of serum LH and FSH levels were similar to those found in bilateral castrates. The levels of GDNFmRNA expression were lower than those in normal controls. On 14th day postoperatively, the spermatogenesis of allotransplanted testes was still not recovered and the expression of GDNFmRNA declined further. CONCLUSION: The atrophy and reduced number of the sertoli cells and the breakage of the close connection probably are the main causes of dysfunction of spermatogenesis. The decline of GDNFmRNA expression is in accordance with the dysfunction of the sertoli cells and the spermatogenesis.

[RNA interference inhibits VEGF expression and growth of PC-3 in prostate carcinoma].

OBJECTIVE: To construct a eukaryotic expression vector carrying human VEGF RNAi and to study the effect of RNA interference on VEGF expression in prostate carcinoma. METHODS: VEGF RNAi was synthesized, inserted into the RNA interference eukaryotic expression vector, and confirmed by the result sequencing. The vector was transfected into prostate cancer PC-3, the VEGF expression detected by Western blot and the cell inhibiting rate determined by MTT. RESULTS: The VEGF RNAi eukaryotic expression vector was successfully constructed. Compared with the empty vector group and the control group, the amount of VEGF protein expression was obviously decreased in the VEGF RNAi group. The inhibiting rates were 23.5% , 33. 5% and 40. 8% at 24, 48 and 72 h respectively. CONCLUSION: VEGF RNAi can inhibit the protein expression and growth of PC-3, which provides an experimental base for the biological therapy of prostate cancer.

[The specific expression profile of homeobox gene NKX3.1 mRNA and protein in prostate tissues and its relationship with primary prostate cancer].

OBJECTIVE: To determine expressions of NKX3.1 mRNA and protein in prostatic tissues and to investigate the relation between homeobox gene NKX3.1 and prostatic carcinoma. METHODS: 76 prostatic tissues (32 cancer, 12 normal prostate and 32 benign prostatic hyperplasia tissues) and 96 non-prostatic tissues were analyzed for the detection of expressions of NKX3.1 mRNA and protein by using semi-quantitative RT-PCR, Western blotting and immunohistochemical technique. RESULTS: In 76 prostatic tissues, NKX3.1 mRNA was detected in 75 specimens (98.7%), whereas in 96 non-prostatic specimens, NKX3.1 mRNA was negatively expressed in the tissues of bladder, kidney, liver, intestine, fat and skin, except for two expressed in testis and one in mammary gland. The expression ratio of NKX3.1 protein in the epithelia cells of prostate was 100%, but in testis mammary gland was 16.7%, in bladder and intestine was 8.3%, and in kidney, liver, fat and skin was 0% (P < 0.01). The total strong positive ratio of NKX3.1 protein in the epithelia cell of prostate was 94.7%, 5.3% in the stroma cell of prostate (P < 0.01), and 13.6% in benignant prostate cell, 40.6% in prostate cancer (P < 0.01), respectively. CONCLUSION: It is suggested that NKX3.1 is not only the prostate-specific homeobox gene, but is the epithelia-cell-specific gene of prostate. It may play an important role in the development of prostatic carcinoma.

[Ultrastructural changes of the rat convoluted seminiferous tubule-after alcohol consumption].

OBJECTIVE: To study the ultrastructural changes of the rat convoluted seminiferous tubule after alcohol consumption. METHODS: Forty-eight Wistar mature male rats were divided into two groups randomly: control group (A) and experimental one (B). 6 ml/(kg x d) of 50 degrees alcohol was perfused through the gastric tube for 39 days in Group B; and 6 ml/(kg x d) of normal saline was supplemented in Group A. The ultrastructure of the rat convoluted seminiferous tubule was observed by transmission electron microscope at day 14, 27 and 40. RESULTS: In Group A, the pykno-basement membrane was unstriated and uniform, Sertoli cells showed cytoplasmic profusion, with big nucleus, well-distributed nucleoplasm, distinct nucleolus, more mitochondria and plain hierarchical tight-junction. And the ultrastructure of the rat convoluted seminiferous tubule in Group B began to change at the end of the first spermatogenic cycle (D 14) and changed more and more evidently with the ethanol administration, mainly as follows: (1) more lysosomes and vacuolisation found in Sertoli cells, and organelles decreased and blurry; (2) more and bigger vacuoles among the spermatogonia, Sertoli cells and basement membrane; (3) obvious apoptosis of spermatogonia and apoptotic bodies aggregated near the membrane; (4) more cytoplasm and vacuolisation in the sperm of the convoluted seminiferous tubule, and disarranged, deleted or clustered mitochondria in the sperm tail; (5) blurry and rigid tight-junction; (6) thickened, wrinkled or broken basement membrane and under-basement CONCLUSION: Alcohol can cause ultrastructural changes of the basement membrane, tight-junction and Sertoli cells of the membrane. rat convoluted seminiferous tubule and apoptosis of spermatogonia.

[Distribution features of Gleason score and its relationship with clinical stages in 346 prostate cancer patients].

OBJECTIVE: To analyze the distribution features of Gleason score and evaluate the relationship between Gleason score and clinical stages in patients with prostate cancer. METHODS: Surveys were made of the inpatients with prostate cancer diagnosed by pathology from January 1992 to June 2005 in our hospital. Gleason score and clinical stages were determined on the basis of pathological examination and clinical data of the prostate cancer patients. The patients were divided into three groups (1992-1999, 2000-2002 and 2003-2005). The Chi-square test was used to evaluate the distribution and differences of Gleason score among the three groups. Spearman rank correlation was applied to the evaluation of the relationship between Gleason score and clinical stages. RESULTS: We found a statistically significant shift in the distribution of Gleason score (chi2 = 17.703, P < 0.01), and a slight increase in the mean Gleason score. The proportion of moderately differentiated tumor increased (chi2 = 10.736, P < 0.01). There was little change in the proportion of Gleason score 7, 8, 9 and 10 (chi2 = 4.038, P > 0.05). Gleason score had a significant positive correlation with clinical stages in the 346 cases of prostate cancer (r = 0.452, P < 0.01). Significant difference was observed between Gleason score 2-6 and 7 or 8-10 (chi2 = 8.786, P < 0.01, chi2 = 22.956, P < 0.01), but not between the latter 2 groups (chi2 = 0.787, P > 0.05) in prediction of organ-confined disease. CONCLUSIONS: Gleason score 7 shows the similar value to Gleason score 8-10 in predicting the progression of the disease. Gleason score was significantly correlated with clinical stages, which suggests that Gleason score is also an important indicator for the prognosis of prostate cancer.

A large intrascrotal calculus.

A large stone with 8.7 cm multiply 7.2 cm multiply 6.5 cm in size and 420 g in weight dropped down spontaneously from a 93-year-old man's scrotum, who had suffered from left intrascrotal mass and pain for more than 20 years. The component of the stone was magnesium ammonium phosphate. To the best of our knowledge, it is the largest intrascrotal calculus reported in the world. We hereby present the case and discuss the diagnosis and etiology of scrotal calculi.