Search

Pantoea ananatis is a bacterium that is found in many agronomic crops and agricultural pests. Here, we isolated a P. ananatis strain (Lstr) from the rice planthopper Laodelphax striatellus, a notorious pest that feeds on rice plant sap and transmits rice viruses, in order to examine its genome and biology. P. ananatis Lstr is an insect symbiont that is pathogenic to the host insect and appears to mostly inhabit the gut. Its pathogenicity thus raises the possibility of using the Lstr strain as a biological agent. To this end, we analysed the genome of the Lstr strain and compared it with the genomes of other Pantoea species. Our analysis of these genomes shows that P. ananatis can be divided into two mono-phylogenetic clades (clades one and two). The Lstr strain belongs to clade two and is grouped with P. ananatis strains that were isolated from rice or rice-associated samples. A comparative genomic analysis shows that clade two differs from clade one in many genomic characteristics including genome structures, mobile elements, and categories of coding proteins. The genomes of clade two P . ananatis are significantly smaller, have much fewer coding sequences but more pseudogenes than those of clade one, suggesting that clade two species are at the early stage of genome reduction. On the other hand, P. ananatis has a type VI secretion system that is highly variable but cannot be separated by clades. These results clarify our understanding of P. ananatis ’ phylogenetic diversity and provide clues to the interactions between P. ananatis , host insect, and plant that may lead to advances in rice protection and pest control.

Uploaded files are raw data and supporting information showing diversity of mcirobes in small brown planthopper.&nbsp

We present the pion mass and decay constant using the overlap fermion valence on domain wall (DW) fermion sea at several lattice spacings. The mixed action effect in the lattice calculation is also studied, and the result suggests that the mixed action effect with the overlap valence on DW sea would be proportional to the fourth power of the lattice spacing. We obtain the pion decay constant at the physical pion mass and $N_f=2$ chiral limit to be $92.4(3)(2)~{\rm MeV}$ and $87.0(5)(7)~{\rm MeV}$ respectively; and the physical u/d averaged quark mass at $\overline{\textrm{MS}}$ 2 GeV is $3.74(4)(5)(5)(3)~{\rm MeV}$ with the linear ${\cal O}(a^2)$ continuum extrapolation. Using the FLAG value of the NLO low energy constant, we obtain the $N_f=2$ chiral condensate to be $\Sigma^{\overline{\textrm{MS}}(2~{\rm GeV})}=\big(266(2)(1)~{\rm MeV}\big)^3$.

To identify complementary laboratory indices for determining the disease status of patients with hepatitis B virus. Subjects were divided into six groups: hepatitis B virus carrier, mild chronic hepatitis B, moderate chronic hepatitis B, severe chronic hepatitis B, fulminant hepatitis B and healthy controls. Serum alanine aminotransferase, total bilirubin and direct bilirubin were measured by an automatic analyser. The levels of T-cell immunoglobulin domain and mucin-domain-containing molecule-3, macrophage inflammatory protein 2, neutrophil gelatinase-associated lipocalin and inducible nitric oxide synthase were measured by ELISA. T-cell immunoglobulin domain, mucin-domain-containing molecule-3, macrophage inflammatory protein 2 and inducible nitric oxide synthase levels were significantly higher in patients with severe chronic hepatitis B compared with those in patients with mild and moderate chronic hepatitis B or fulminant hepatitis B (P < 0.05). When normal or abnormal alanine aminotransferase was present, significant differences between macrophage inflammatory protein 2 and T-cell immunoglobulin domain and mucin-domain-containing molecule-3 levels between patients with mild, moderate, severe chronic hepatitis B or fulminant hepatitis B were observed (P < 0.05). Our results suggest that T-cell immunoglobulin domain and mucin-domain-containing molecule-3 and macrophage inflammatory protein 2 could serve as alanine aminotransferase, direct bilirubin or total bilirubin complementary indices for determining the status of patients with hepatitis B.

In the present study, we report the case of a 69-year-old female who developed urinary leakage following partial nephrectomy (PN) to remove left renal masses. The results of CT and MR urography revealed left proximal ureteral obstruction and urinary fistula. Reoperation was performed on the 16th postoperative day to explore the left kidney and ureter in order to relieve the obstruction. The left proximal ureter was found to be enfolded by fibrin glue and showed marked stiffness and adhesion during the reoperation. The lesion of the ureter was resected and the ureter was anastomosed with the routine double-J stent. Pathological examination of surgical specimens revealed fat fibrous scar tissue hyperplasia with inflammatory cell infiltration. The patient recovered completely without exudate. Our experience suggests that care should be taken to avoid touching the ureter with fibrin glue during PN surgery.

Summary. A mammalian expression vector comprised of the PreS2-TLM (translocation motif), a single-chain variable fragment (ScFv) that binds to hepatitis B surface antigen (HBsAg) and the EGFP gene was constructed. A stably transformed cell line that could express and secrete the fusion protein (PreS2-TLM-ScFv-EGFP) was established. HBsAg-positive HepG2.2.15 cells and HepG2 and HeLa cells were incubated with the supernatant of the transformed cell line cultures for evaluating the cellular permeability of PreS2-TLM-ScFv-EGFP. The location of the fusion protein PreS2-TLM-ScFv-EGFP in HepG2.2.15 cells was observed with immunofluorescence staining. EGFP was next replaced by a dominant negative mutant of the hepatitis B virus core gene (HBcDN) for producing fusion protein PreS2-TLM-ScFv-HBcDN, which was detected by western blot. The supernatant containing fusion protein PreS2-TLM-ScFv-HBcDN was added to the cultures of HepG2.2.15 cells, and the packaged hepatitis B virus (HBV) pregenomic RNA expression levels in the cells were measured using qRT-PCR. The results of the in vitro study indicated that the packaged HBV pregenomic RNA expression levels in HepG2.2.15 cells significantly decreased when these cells were exposed to the supernatant at the dose of 25% for 24, 48 and 72 h, or at the dose of 12.5% for 72 h.

We investigated the inhibitory effect of small interfering RNA (siRNA) targeting survivin gene on cell proliferation and apoptosis in human renal clear cell carcinoma 786-O cells. qRT-PCR, immunocytochemistry, and Western Blot were used to detect Survivin gene expression in 786-O cells. Cell proliferation was determined by BrdU assay and PCNA expression. Cell apoptosis was analyzed through detection of caspase-3 activity, and the effect of survivin-siRNA on Bcl-2 gene expression was also examined. Forty-eight hours after transfection, survivin expression was markedly inhibited at the mRNA and protein level. Downregulation of survivin resulted in a significant inhibition of tumor cell growth. Caspase-3 activity showed that siRNA targeting survivin gene induced cell apoptosis in 786-O cells. Moreover, Bcl-2 protein expression was markedly inhibited by transfection with siRNA against survivin. These results indicate that siRNA targeting survivin gene can downregulate survivin gene expression in 786-O cells, inhibit growth, and induce apoptosis of renal carcinoma cells.

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.

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.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.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.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.

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.

Objective To clarify the effects of androgens (testosterone and dihydrotestosterone, DHT) and ageing on gene expression of vasoactive intestinal polypeptide (VIP), assessed as the expression of VIP mRNA, in rat corpus cavernosum. Materials and methods The study comprised 160 male Sprague Dawley rats divided into group A (56 rats, 5 weeks old), group B (50 rats, 10 weeks old) and group C (54 rats, 58 weeks old). Groups A–C were subdivided, respectively, into subgroups 1 (intact controls), 2 (castrated), 3 (castrated but given testosterone undecanoate 25 mg/kg per month by intramuscular injection), 4 (castrated with testosterone undecanoate 50 mg/kg per month) and 5 (treated with finasteride 4.5 mg/kg per day, orally). At 4 and 10 weeks after these treatments half the rats were killed. Serum samples were taken for the measurement of total and free testosterone and DHT, using a radioimmunoassay. Penile samples (corpus cavernosum) were frozen in liquid nitrogen and stored at −80°C. VIP mRNA was estimated using a semi-quantitative reverse-transcription polymerase chain reaction. Results There was no significant difference in VIP mRNA in rat corpus cavernosum between intact control and castrated subgroups, or subgroups treated with finasteride, in groups A–C, including both the 4- and 10-week old animals (P > 0.05). Penile VIP mRNA was unchanged at any dose of testosterone in the castrated subgroups in all groups (P > 0.05). There was no significant relationship between penile VIP mRNA and ageing (P > 0.05). Conclusions The gene expression of VIP in rat corpus cavernosum is independent of androgens (testosterone and DHT) and ageing. Androgens probably induce penile erection by pathways other than VIP; ageing may have little effect on penile VIP mRNA.

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 6week-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. (Asian J Androl 2006 Sep; 8: 569–575)

To determine the feasibility of transfecting penile corpora cavernosa with pcDNA3/vasoactive intestinal polypeptide (VIP) cDNA, which encodes for VIP in streptozotocin (STZ)-diabetic rats, to clarify whether transfection of VIP cDNA into the cavernosum affects the physiological response to cavernosal nerve stimulation, and whether this process would affect other organs in the diabetic rat model in vivo.pcDNA3/VIP cDNA was injected into the corpus cavernosum of STZ-induced diabetic Sprague-Dawley rats. The intracavernosal pressure (ICP) and response to electrical stimulation of the cavernosal nerve (15 Hz, 1.5 ms, 20 V, 1 min) were measured in subsamples of rats at 1, 3, 7 and 14 days after injection; after measuring the ICP the animals were killed, and penile, hepatic, renal artery and abdominal aorta tissue samples were frozen in liquid nitrogen and stored at -80 degrees C. The gene expression of VIP in all samples, assessed as the expression of VIP mRNA, was estimated using a semiquantitative reverse-transcription polymerase chain reaction.The mean amplitude of ICP and expression of VIP mRNA in the cavernosa of the VIP-treated rats was greater at 1, 3, 7 and 14 days after injection (P0.05) than in the control animals. There were no changes in the expression of VIP mRNA in hepatic, renal and abdominal aorta samples after injection (P0.05).VIP cDNA is easily incorporated into corpus cavernosum, and the expression is sustained foror = 2 weeks in the penis in vivo. The transfer of VIP is capable of altering the physiologically relevant erectile response, as measured by an increase in the ICP after stimulating the cavernosal nerve. The intracorporal micro-injection of pcDNA3/VIP cDNA had little effect on the expression of VIP mRNA in other important organs.

To study the androgen dependence of the neurotransmitter, calcitonin gene-related peptide (CGRP) in rat penis.Forty-four Sprague-Dawley rats were randomly divided into Group A (intact controls), Group B (castrated) and Group C (gavaged with finasteride 4.5 mg.kg(-1).day(-1)). Four and ten weeks later respectively, half of rats in each group were anaesthetized. Blood samples were taken for the measurement of serum testosterone and dihydrotestosterone (DHT) by means of radioimmunoassay. Penile samples were harvested for the investigation of calcitonin gene related peptide (CGRP)-immunoreactive nerve fibers with immunohistochemistry. The computer-assisted imaging analysis system was applied to calculate the area proportion of the CGRP-positive nerve fibers (CGRP-PNF) in each group.1) Both 4 and 10 weeks later, testosterone and DHT levels in Group B decreased significantly compared with those in Group A, (P0.05, P0.01, respectively); DHT level in Group C was also significantly decreased in comparison with that in Group A for both 4- and 10- week animals (P0.05); 2) There was no significant differences in area proportion of CGRP-PNF among Groups A, B and C 4 weeks after treatments (P0.05); However, 10 weeks later, the proportion of CGRP-PNF in Groups B and C was significantly less than that in Group A (P0.01); 3) The proportion of CGRP-PNF of 4-week animals in Groups B and C was significantly higher than that of 10-week animals (P0.05).The expression of neurotransmitter, CGRP may depend on androgens, including testosterone and DHT in rat penis.

Nephrogenic diabetes insipidus (NDI) with dilatation of upper urinary tract and bladder is rarely reported. This study evaluated the urological manifestations and treatment results of this rare condition.According to a fluid deprivation test, seven men and two women were diagnosed as NDI, and underwent ultrasonography, magnetic resonance urography, cystography, and urodynamic and laboratory examination. A micturition diary and symptoms were recorded before and during treatment. All patients were given hydrochlorothiazide and followed up with ultrasonography, the postvoiding residual volume (PVR), urinalysis and kidney function tests.All patients presented with polydipsia and polyuria with severe dilatation of upper urinary tract and bladder. Urodynamics showed that the mean values of the bladder capacity, bladder pressure at the end of filling, maximum flow rate (Q(max)), detrusor pressure and PVR were 1056.3 ml, 40.5 cmH(2)O, 30.8 ml/s, 51.3 cmH(2)O, 436.3 ml, respectively. The total urine volume in all patients decreased by more than 50% after administration of hydrochlorothiazide, which improved the symptoms and hydronephrosis in seven cases. The symptoms of other two cases were relieved after treatment with an electric transurethral incision of the bladder neck and a cystostomy.NDI should be considered in patients with dilatation of the urinary tract and polyuria. Higher bladder pressure at the end of filling may contribute to the dilatation of urinary tract. Normal detrusor contractility with large PVR is a unique manifestation of this condition. Controlling the urine volume and reducing the PVR are of key importance in the treatment of this condition.

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.

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-β1 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-β1 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.

The aim of this study was to investigate the influence of the duration of bladder overdistention (DOBO) on kidney structure and function in rats. Bladder overdistention was induced in male Sprague-Dawley rats by an infusion of saline. Forty rats were divided into five groups: DOBO 1, 2, 4 or 8 h groups and the control. Renal function was evaluated using serum creatinine (SCr) and blood urea nitrogen (BUN). Apoptotic indices and morphologic changes of the kidney were detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL) staining and transmission electron microscopy (TEM). Compared with the control, rats undergoing 2, 4 or 8 h of overdistention showed significant, time-dependent increases in SCr (12.375 vs. 23.125, 34.375 and 51.500 μmol/l, respectively), BUN (6.980 vs. 18.689, 25.184 and 32.079 mmol/l, respectively), renal size (1.041 vs. 1.472, 1.484 and 1.634 cm3, respectively) and renal pelvis separation (0.000 vs. 0.223, 0.320, 0.308 and 0.277 cm, respectively; P

Overexpression of Bcl-2 protein in cancer cells can inhibit programmed cell death and engender chemoresistance. Bcl-2 antisense oligonucleotide (G3139) has shown its antitumor effects enhanced in preclinical models when combined with taxol-based chemotherapy. This study aimed to investigate the efficacy of G3139 combined with epirubicin in the androgen-independent prostate cancer.PC3 prostate cancer cell line was cultured and treated with epirubicin and Bcl-2 antisense oligonucleotide alone or in combination. The effects of therapeutic agents on cells were determined by the MTT assay. Expression of Bcl-2 mRNA and protein was documented by RT-PCR and Western blotting. Apoptosis induction was confirmed by flow cytometric analysis.Bcl-2 antisense oligonucleotide alone produced no cytotoxic effects and the combination of Bcl-2 antisense oligonucleotide with epirubicin sensitized PC-3 cells to the killing effects of chemotherapy. A marked down-regulation of Bcl-2 mRNA and protein was observed after antisense and epirubicin cotreatment. A statistically significantly higher fraction of apoptotic cells was detected by flow-cytometric analysis after epirubicin treatment with prior antisense Bcl-2 transfenction, as compared with mono antisense Bcl-2 or epirubicin treatment.These data suggested that inhibition of Bcl-2 expression combined with epirubicin may be an attractive therapeutic strategy in hormone-refractory prostate cancer.

The effects on sterilization and preservation of chilled pork by electron beam irradiation were studied in this article. The mathematics model was established by response surface analysis, which beam energy(X1) and dose(X2) as factors and colony form unit(Y) as responses. The results showed that the model of electron beam irradiation on sterilization of chilled pork can be expressed Y=3.78-0.24X1-0.13X2 -0.16X1X2-0.18X12+0.15X22(R2=0.9755). At the same time, the interaction between electron beam energy and absorbed doses has been found, and the significance sequence of factors was absorbed dose>;interaction>;electron beam energy. The initial total number of colony would drop 2 logarithm units when absorbed doses were in range from 3.23kGy to 4.0kGy and electron beam energy was in range from 2.3MeV to 3.8MeV. When the samples were stored at 4°C, the shelf life of irradiated samples was 18d. The results also showed that electron beam irradiation has effects on sterilization and preservation of chilled pork, and the application of electron beam irradiation was confirmed very useful for meat industry.

A two-dimensional generalized cubic nonlinear Schr��dinger equation with complex coefficients for the group dispersion and nonlinear terms is used to investigate the evolution of a finite-amplitude localized initial perturbation. It is found that modulation of the latter can lead to side-band formation, wave condensation, collapse, turbulence, and inverse cascade, although not all together nor in that order., 12 pages, 5 figures

To explore the feasibility of inhibition of hTERT and androgen receptor (AR) gene expression simultaneously in LNCaP cells by single shRNA vector.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.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 (P0.05).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.

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.

To determine the effect of MEK inhibitor (U0126) on donor testes from ischemia-reperfusion injury after orthotopic testicular transplantation in rats.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.Group 1, 2 and 3 had no significant differences in all results (P0.05). The levels of ERK1, ERK2, pERK1 and pERK2 in Group 4 were significantly increased compared with Group 1 (P0.05), the levels of ERK1 and ERK2 in Group 6 were not different from those of Group 4 (P0.05), but the levels of pERK1 and pERK2 in Group 6 were lower than those in Group 4 significantly(P0.05), 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 (P0.05) which remained at the normal levels.U0126 has a protective effect on the donor testes in a short period through inhibiting expression of pERK1/2 activated by testicular transplantation.

MicroRNAs (miRNAs) have recently been shown to down-regulate gene expression by targeting mRNA translation and to play a critical role in tumorigenesis; how they regulate bladder tumor development, particularly in patients, is, however, poorly understood. The difference in miRNA expression in a bladder tumor compared with healthy tissue from the same patients was examined using microRNA arrays in seven patients. Here, we showed that up-regulation of miRNA was not commonly found in this limited number of patients, and four miRNAs (miR-26a, miR-29c, miR-30c, miR-30e-5p) were down-regulated as a common marker in patients with a 1-3 grade of disease. Our data suggest that instead of up-regulation of carcinogenic miRNAs, loss of regulation of these miRNA may be critical for bladder tumor development in patients.

To introduce a method of retrograde ureteric catheter placement via flexible cystoscope , and to evaluate the feasibility and safety of this method.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.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, P0.05]. 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, P0.05]. However, there were not significant differences in the inserting times in both men and women between these 2 groups (both P0.05). 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 P0.05). 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, P0.05). Four patients had a fever after flexible cystoscopy while 6 cases did after rigid cystoscopy, however, without significant difference between these 2 groups (P0.05).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.

To investigate the utility of prostate specific antigen density for detecting prostate cancer in men with serum PSA levels of 4-10 ng/mL.Between January 2003 and November 2007, 237 men (aged 48-84 years, median 71) with total PSA levels of 4-10 ng/mL participated in a protocol for prostate cancer screening. Eligible patients were recommended for transrectal ultrasonography (TRUS)-guided prostate biopsies after measuring prostate volumes transrectally. The diagnostic value of PSA levels and the free-to-total PSA ratio (f/tPSA), PSA densities (PSAD) were compared using receiver operating characteristic analysis.Prostate cancer was diagnosed in 44 (18.6%) of the 237 men who had biopsies. There were significant differences between the groups in the prostate volumes determined by TRUS, PSAD, PSA levels and f/tPSA, whereas there was no significant difference in patient age. The area under the curve (AUC) of PSA (0.6786) and PSAD (0.717) was similar and significantly greater than that of f/tPSA (AUC 0.329). PSAD was a significantly better indicator of prostate cancer than f/tPSA. The sensitivity and specificity of PSA density at a cutoff of 0.134 ng/mL(2) was 90 and 33.7%, respectively.PSAD was a better predictor of prostate cancer in Chinese men with PSA levels of 4-10 ng/mL, especially those who have had prior ultrasound-determined measurements of prostate volume. Our data suggest that different PSAD cutoffs may need to be defined for Chinese.

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.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.Comparisons of the normal, hyperplastic and tumour prostate tissues indicated an overexpression of ERalpha in tumour specimens (P0.01). However, the expression of ERbeta significantly reduced in tumour tissues compared with normal and hyperplastic specimens (P0.01), 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 P0.05), and a positive correlation between ERalpha expression and tumour stage, grade (0.51, 0.57, respectively, both P0.01). Our analysis also showed that hormone refractory, prostate cancer, compared with hormone dependent, prostate cancer, displayed a decreased expression of ERbeta (P0.01) and an increased expression of ERalpha.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 (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.

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.

To construct a eukaryotic expression vector carrying human VEGF RNAi and to study the effect of RNA interference on VEGF expression in prostate carcinoma.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.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.VEGF RNAi can inhibit the protein expression and growth of PC-3, which provides an experimental base for the biological therapy of prostate cancer.

To analyze the distribution features of Gleason score and evaluate the relationship between Gleason score and clinical stages in patients with prostate cancer.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.We found a statistically significant shift in the distribution of Gleason score (chi2 = 17.703, P0.01), and a slight increase in the mean Gleason score. The proportion of moderately differentiated tumor increased (chi2 = 10.736, P0.01). There was little change in the proportion of Gleason score 7, 8, 9 and 10 (chi2 = 4.038, P0.05). Gleason score had a significant positive correlation with clinical stages in the 346 cases of prostate cancer (r = 0.452, P0.01). Significant difference was observed between Gleason score 2-6 and 7 or 8-10 (chi2 = 8.786, P0.01, chi2 = 22.956, P0.01), but not between the latter 2 groups (chi2 = 0.787, P0.05) in prediction of organ-confined disease.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.

Transitional cell carcinoma (TCC) of the urothelium is often multifocal and subsequent tumors may occur anywhere in the urinary tract after the treatment of a primary carcinoma. Patients initially presenting a bladder cancer are at significant risk of developing metachronous tumors in the upper urinary tract (UUT). We evaluated the prognostic factors of primary invasive bladder cancer that may predict a metachronous UUT TCC after radical cystectomy. The records of 476 patients who underwent radical cystectomy for primary invasive bladder TCC from 1989 to 2001 were reviewed retrospectively. The prognostic factors of UUT TCC were determined by multivariate analysis using the COX proportional hazards regression model. Kaplan-Meier analysis was also used to assess the variable incidence of UUT TCC according to different risk factors. Twenty-two patients (4.6%). developed metachronous UUT TCC. Multiplicity, prostatic urethral involvement by the bladder cancer and the associated carcinoma in situ (CIS) were significant and independent factors affecting the occurrence of metachronous UUT TCC (P = 0.0425, 0.0082, and 0.0006, respectively). These results were supported, to some extent, by analysis of the UUT TCC disease-free rate by the Kaplan-Meier method, whereby patients with prostatic urethral involvement or with associated CIS demonstrated a significantly lower metachronous UUT TCC disease-free rate than patients without prostatic urethral involvement or without associated CIS (log-rank test, P = 0.0116 and 0.0075, respectively). Multiple tumors, prostatic urethral involvement and associated CIS were risk factors for metachronous UUT TCC, a conclusion that may be useful for designing follow-up strategies for primary invasive bladder cancer after radical cystectomy.

To study the ultrastructural changes of the rat convoluted seminiferous tubule after alcohol consumption.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.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-basementAlcohol can cause ultrastructural changes of the basement membrane, tight-junction and Sertoli cells of the membrane. rat convoluted seminiferous tubule and apoptosis of spermatogonia.

To establish the testis transplantation model in rats and to study the mechanism of graft injury.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.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.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.

SUMMARY: Background: PI3K/Akt pathway has been shown to play a critical role in the regulation of mitogenic signalling, apoptosis, cell proliferation and survival in a variety of cells and tissues. The aim of the present study was to investigate the role of PI3K/Akt pathway in the renal ischaemia/reperfusion. Methods: Four experimental groups, sham-operative mice, vehicle-delivered and wortmannin-treated ischaemic/reperfusion injury mice, wortmannin-treated normal mice were designed to examined serum blood urea nitrogen level, renal injury, proliferating cell nuclear antigen protein and Akt phosphorylation status at 30 min, 90 min, 24 h, 48 h of reperfusion after ischaemic treatment. Wortmannin or its vehicle was given intraperitoneally at 4 h before surgery. Blood urea nitrogen was measured, and immunohistochemistry and western blotting were used to detect the components of PI3K/Akt pathway in the ischaemic/reperfusion injury kidney. Results: PI3-kinase inhibitor wortmannin imposes a deleterious effect on serum blood urea nitrogen level, renal function after renal ischaemia/reperfusion injury in mice. The renal cell proliferation increased after ischaemia/reperfusion injury in mouse, which could be inhibited by wortmannin. Phosphorylation of Akt was increased after ischaemia/reperfusion in the mouse kidney, and reduced by wortmannin administration. Conclusion: This primary study suggests that PI3-kinase/Akt signalling pathway play an important role in the regulation of the renal repair after ischaemia/reperfusion injury.

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.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.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% (P0.01). 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 (P0.01), and 13.6% in benignant prostate cell, 40.6% in prostate cancer (P0.01), respectively.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.

To establish a model of testis transplantation in the rat using cuff techniques.Sixty healthy inbred Wistar rats were included as donors and recipients. The testicular artery with the aortic segment with two cuffs was anastomosed to the recipient's aorta and the testicular vein with a cuff was anastomosed to the left common iliac vein of the recipient. The vas deferens was end-to-end anastomosed.Of the 30 rats undergoing transplantation, 27 survived, 2 died from paralytic intestinal obstruction and 1 died from hemorrhage. Of the 27 survivals, 4, 4 and 5 were vivisected on the 14th, 28th and 56th day after operation, respectively, showing a good blood supply to the graft. Another 14 rats were used in another experiment, which revealed no obvious congestion, necrosis and edema in the left lower limb of the recipients.The model of allogeneic rat testis transplantation established with cuff techniques, simple, stable and reliable, can be used as a tool for the study of testis transplantation.

To investigate the age and pathological features of prostate cancer patients in recent years.An analysis was made of the age and pathological features of 481 cases of prostate cancer pathologically diagnosed from January 1998 to April 2004, 39 cases in 1998, 69 in 1999, 73 in 2000, 68 in 2001, 72 in 2002, 121 in 2003, and 39 in the first four months of 2004.The patients ranged in age from 40 to 91 years, averaging 72, 95% between 55 and 84, and 84.2% over 65 years. Pathologically, 14 cases were well, 29 moderately, and 83 poorly differentiated according to the three-grade system (WHO, the Mostofi system), with 355 cases ungraded. Forty cases (8.3%) were microcarcinoma (1 cm), and 20 cases (4.2%) incidental carcinoma. Of the total number, 473 cases (98.1%) were pathologically diagnosed as adenocarcinoma, 1 endometrioid adenocarcinoma, 1 squamous cell carcinoma, 1 signet ring cell carcinoma, 1 adenosquamous cell carcinoma, 1 small cell carcinoma, 1 mucinous adenocarcinoma, 1 adenoid cystic carcinoma, and 1 transitional cell carcinoma.Prostate cancer commonly develops in men over 65 years, and adenocarcinoma is the most common histological type. The disease has become a major malignant tumor to endanger elderly males.

To clarify the ultrastructural changes of penile tunica albuginea (TA) in streptozotocin (STZ)-induced diabetic rats.Intraperitoneal injection of STZ was used to induce diabetes mellitus (DM) in 12 Sprague Dawley rats. Ten rats (age and weight-matched) were used as control. Blood samples from the tail snips of the rats were used for the determination of serum glucose levels with SureStep Plus Blood Meter. At week 4 and 10 after the injection, half of the rats in each group were sacrificed and penile samples were obtained from the middle third of the penile shaft for the examination of TA under scanning electron microscopy.In the diabetic group, the serum glucose levels were higher (P0.01 at both time points) and the TA were thinner (P0.05) than those of the controls. In the control group, the fibers of TA were rich and arranged regularly and undulated, while in the diabetic group, the fibers were diminished, lost the undulations and were arranged irregularly.In rats, DM appeared to impair the penile TA ultrastructures and this impairment could contribute to diabetic erectile dysfunction in part by impairing the veno-occlusive function.

To determine the expression of estrogen receptor-alpha (ERalpha) and estrogen receptor-beta (ERbeta) in prostatic carcinoma (PCa).The expression of ERalpha and ERbeta was analysed in 32 cases of PCa, 12 cases of normal prostate tissue and 32 cases of benign prostate hyperplasia (BPH) by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR), and the genes were sequenced.Compared with the tissue of BPH, the ERalpha expression significantly increased, but the ERbeta expression decreased in the tissue of PCa (P0.01). Compared with in the early stage and high differentiation of prostatic carcinoma, the ERalpha expression increased obviously, but ERbeta expression decreased in the developed stage and low differentiation (P0.01). ERalpha increased, but ERbeta decreased in hormone-refractory prostatic carcinoma (P0.05).ERalpha and ERbeta may play an important role in the development of PCa. It is shown that analysis of the expression of ER, especially ERbeta in PCa, will benefit to the diagnosis and treatment of this disease.

To study significance and limitations of the ratio of free to total prostate specific antigen (f/t PSA) in differential diagnosis between prostate cancer and benign prostatehyperplasia (BPH) with total PSA (tPSA) levels between 4 and 10 ng/ml.We analysed retrospectively 180 prostate cancer and BPH patients who were diagnosed and treated in our hospital from October 1998 to October 2002 and had serum tPSA levels between 4 and 10 ng/ml. Of the 180 patients, 36 (20%) were histologically confirmed as prostate cancer and 144 (80%) BPH. The tPSA and free PSA (fPSA) in serum were measured by micropartical enzyme immunoassay. Prostate volume was measured by transabdominal ultrasonography. We chose Student's t-test for comparison between prostate cancer and BPH groups. The correlation between prostate volume and f/t PSA was analyzed using Pearson's correlation coefficient.The mean values of tPSA and f/t PSA were 6.75 ng/ml and 0.17 in patients with prostate cancer, 6.48 ng/ml and 0.25 in patients with BPH. The mean value of tPSA wasn't significantly different between patients with prostate cancer and BPH (P0.05). However, the mean value of f/t PSA of patients with prostate cancer was significantly lower than that of patients with BPH (P0.01). Furthermore, there were significant and positive correlation between prostate volume and f/t PSA in both groups with prostate cancer and BPH (prostate cancer group's correlation coefficient (r = 0.50, P0.01); BPH group (r = 0.24, P0.01). There was significant difference in f/t PSA between prostate cancer and BPH patients with prostate volumes more than 40 cm(3) (P0.05), but not between these two groups with prostate volumes more than 40 cm(3) (P0.05).The f/t PSA is significant in differential diagnosis between prostate cancer and BPH with tPSA levels between 4 and 10 ng/ml. But prostate volume has an effect on f/tPSA. The f/tPSA has diagnostic value of differentiation only when the prostate volume is less than 40 cm(3).

To evaluate the effect of pretreatment with finasteride in decreasing intraoperative bleeding and irrigating fluid absorption during transurethral resection of prostate (TURP).Eighty patients with benign prostate hypertrophy undergoing TURP were divided into two groups: 40 patients were pretreated with finasteride for 7 to 14 days before TURP and 40 patients without pretreatment. Absorption of irrigating fluid was quantified by analyzing the serum concentration of gentamycin. Intraoperative blood loss was calculated based on hemoglobin concentrations before and after operation.The whole blood loss, hemoglobin concentration of irrigating fluid used, blood loss per minute, blood loss per gram tissue resected, whole irrigation absorption, irrigation absorption per minute and per gram tissue resected in patients pretreated with finasteride were significantly less than those in patients without pretreatment (P0.05). The blood transfusion volume, the incidence of hypotension and hyponatremia in patients pretreated with finasteride were significantly less than those in patients without pretreatment (P0.05).Pretreatment with finasteride is of value in reducing intraoperative bleeding, irrigation absorption and perioperative complication during TURP.

To investigate whether different types of injury on bladder wall can influence bacillus Calmette-Guerin (BCG) attachment.The bladder mucosa of 24 rabbits were treated by electrocautery,cryocautery and incision on left lateral wall, right lateral wall and posterior wall, respectively. Then radiolabeled BCG ((3)H-BCG) was instilled into bladder. Two hours latter, the injured bladder wall with different methods and non-injured wall (anterior wall of bladder) were surgically removed and digested. The quantity of BCG of each specimen was determined by liquid scintillation counter.The quantity of BCG attachment to bladder wall with different injuries was significantly higher than that of non-injured wall (P0.001), meanwhile there was no statistically difference among the BCG levels of different injury types (P0.05).BCG attachment is not influenced by different types of injury on the bladder wall.

The F-box protein Skp2 is required for the ubiquitin-mediated proteolysis of the cyclin-depended kinase inhibitor p27(kip1). Overexpression of Skp2 has been reported in many cancers, including breast carcinoma, gastric carcinoma, and prostate cancer. The purpose of this study was to investigate the correlation of Skp2 and p27(kip1) expression with the clinicopathological features of prostatic carcinoma, and the correlation between expression of Skp2 and p27(kip1) in prostate cancer.Skp2 and p27(kip1) protein expression were evaluated in the tissues of 41 human prostatic carcinomas as well as 20 benign prostatic hyperplasia (BPH) using immunohistochemistry (EnVision method).The Skp2 labeling frequency in prostatic carcinoma (8.52%+/-2.40%) was significantly higher than that in BPH (0.21%+/-0.15%)(P0.001). The Skp2 protein expression in prostatic carcinoma was positively correlated with preoperative serum prostate-specific antigen level (r=0.360,P=0.021), extraprostatic extension (r=0.570,P0.001), tumor stage (r=0.531, P0.001), and histological grade (r=0.514,P=0.001). The p27(kip1) labeling frequency in prostatic carcinoma(70.71%+/-4.25%) was significantly lower than that in BPH (97.21%+/-2.10%) (P0.001). The p27(kip1) protein expression in prostatic carcinoma was inversely correlated with preoperative serum prostate-specific antigen level (r=-0.399,P=0.010), extraprostatic extension (r=-0.329, P=0.036), tumor stage (r=-0.453,P=0.003), and histological grade (r=-0.290,P=0.046). Skp2 expression was inversely correlated with p27(kip1) in prostate cancer (rho=-0.572,P0.001).Expression of Skp2 protein may lead to decrease p27(kip1) level in human prostatic carcinoma, indicating its involvement in the development of human prostatic carcinoma. It may provide new targets for the therapy of prostate cancer.

Trauma, tumor, disease and congenital abnormalities may lead to genital organ damage or function failure, and consequently the requirement of its reconstruction. Tissue engineering follows the principles of cell transplantation, materials science and engineering toward the development of biological substitutes that would restore and maintain normal function. These new techniques have been recently introduced into the field of andrology. Based on the latest advances, the present paper afferds a general prospect of the future direction of the development of tissue engineering in andrology.