[Low-dose testosterone undecanoate capsules combined with tadalafil for late-onset hypogonadism accompanied with ED].

OBJECTIVE: To observe the clinical effect of low-dose testosterone undecanoate capsules combined with tadalafil on late-onset hypogonadism (LOH) accompanied with ED. METHODS: Ninety cases of LOH accompanied with ED who met the inclusion criteria were randomly divided into a control group and a combination therapy group, the former treated with tadalafil and the latter with low-dose testosterone undecanoate capsules combined with tadalafil. The LOH symptoms, IIEF-5 scores, sexual encounter profile (SEP) scores, prostate volumes, and the levels of total testosterone (TT), free testosterone (FT) and prostatic specific antigen (PSA) were recorded and compared between the two groups before and after treatment. RESULTS: The IIEF-5 and SEP scores and the TT and FT levels were 20.6 +/- 3.8, 4.02 +/- 1.08, (15.4 +/- 3.4) nmol/L and (0.391 +/- 0.062) nmol/L, respectively, in the combination therapy group after treatment, significantly higher both than 15.7 +/- 3.9, 1.49 +/- 0.82, (10.1 +/- 1.2) nmol/L and (0.200 +/- 0.045) nmol/L before treatment (P < 0.01) and than 8.6 +/- 3.6, 3.50 +/- 1.21, (10.2 +/- 1.2) nmol/L and (0.210 +/- 0.051) nmol/L in the control group after treatment (P < 0.01). CONCLUSION: Low-dose testosterone undecanoate capsules combined with tadalafil has a definite clinical effect and no obvious adverse reactions in the treatment of LOH accompanied with ED.

[Role of beta-catenin signaling pathway in EMT of human prostate cancer induced by HIF-1alpha].

OBJECTIVE: Epithelial-mesenchymal transition (EMT) is an important process in tumor development. Several studies suggest that the beta-catenin signal pathway may play an important role in EMT. However, there is no direct evidence showing that this pathway actually determines the EMT induced by exogenous signal. Our previous study has successfully proved that over-expression of HIF-1alpha could induce EMT in LNCaP cells, but not in PC-3. So the present study was intended to indicate that the signal of HIF-1alpha for inducing prostate cancer cell to undergo EMT might pass through the beta-catenin pathway. METHODS: Firstly, we analyzed the expression of HIF-1alpha and its target proteins in LNCaP/HIF1alpha and PC-3/HIF1alpha by Western blot. And then EMT-associated proteins were detected by Western blot. Furthermore the potency of invasiveness and proliferation of several cell lines were evaluated by transwell and MTT assay. Lastly the expressions of beta-catenin and GSK-3beta in these cells were analyzed by Western blot and RT-PCR. RESULTS: HIF-1alpha, Glut-1 and VEGF were highly expressed in LNCaP/HIF1alpha and PC-3/HIF1alpha. And PC-3, LNCaP and PC-3/HIF1alpha were EMT-negative cell lines whereas LNCaP/HIF1alpha and IA8 had undergone EMT process. LNCaP/HIF1alpha, IA8 and PC-3/HIF1alpha exhibited a much stronger potency of invasiveness and proliferation than those of PC-3 and LNCaP. The protein levels of total GSK-3beta and phospho-GSK-3beta in LNCaP/HIF1alpha, IA8 and PC-3/HIF1alpha cells significantly decreased; however, the relative ratios of p-GSK3beta/t-GSK3beta increased. Consistently, beta-catenin protein expression increased in LNCaP/HIF1alpha and IA8 cells, but not in PC-3/HIF1alpha; RT-PCR confirmed these results, except for an enhanced transcription activity of beta-catenin mRNA in PC-3/HIF1alpha. CONCLUSION: The activation of the beta-catenin signaling pathway correlates with the characteristic of EMT and potency of invasiveness and proliferation. And it may be one critical factor directly controlling the process of EMT induced by HIF-1alpha in prostate cancer.

[Clinical efficacy of Yangjing Decoction on idiopathic asthenozoospermia].

OBJECTIVE: To evaluate the clinical effect of the Chinese medicine Yangjing Decoction on idiopathic asthenospermia. METHODS: This study included 62 patients with idiopathic asthenospermia diagnosed with the computer-assisted semen analysis system and other methods based on the WHO guidelines. The patients were equally randomized to a trial and a control group, the former treated with Yangjing Decoction at the dose of 400 ml bid, and the latter with Yougui Capsules tid, both for a course of 6 months. Then we analyzed the changes in sperm concentration and percentage of grade a + b sperm in the patients, as well as the pregnancy in their wives. RESULTS: Compared with the controls, the patients of the trial group showed a significantly elevated percentage of grade a + b sperm after 6 months medication (P < 0.01), though no statistically significant differences were observed in sperm concentration (P > 0.05). The total rate of effectiveness was 87.09% and 7 pregnancies were achieved in the trial group, as compared with 66.74% and 3 pregnancies in the controls (P < 0.05). CONCLUSION: Yangjing Decoction can significantly improve sperm vitality, and has a desirable effect on idiopathic asthenospermia.

[Over-expression of HIF-1alpha induces EMT of human prostate cancer cells].

OBJECTIVE: To determine whether human prostate cancer cell lines undergo epithelial-mesenchymal transition (EMT) and become more invasive when induced by HIF-1alpha, and to explore the underlying molecular mechanism. METHODS: The cell line LNCaP, appropriate for the HIF-1alpha induction test, was screened out from 4 different EMT-negative prostate cell lines through vimentin gene detection by RT-PCR. The recombinant plasmid pCDNA3. 1(-)/HIF-1alpha was constructed and transfected into LNCaP with the Lipofectamine 2000 system. The control plasmid pCDNA3.1 (-) was transfected by the same method. The positive clone cells were selected by G418 and confirmed by Western blot and immunofluorescence staining. Then a Transwell polycarbonate filter, coated with 100 micol Matrigel at 1:20 dilution in the serum-free medium, was used to analyze the invasive potency. The expression of E-cadherin and vimentin was detected by Western blot. RESULTS: Among the 4 different EMT-negative cell lines, LNCaP was the only one that expressed the vimentin gene but not protein. The expression of HIF1alpha was obviously higher in LNCaP/HIF1alpha than in LNCaP/pCDNA3. 1 (- an LNCaP. The number of the LNCaP/HIF1alpha cells that penetrated through the Transwell polycarbonate filter was significantly larger than that of the LNCaP and LNCaP/pCDNA3. 1(-) cells. Compared with the LNCaP/pCDNA3.1(-) and LNCaP cells, the expression of vimentin was up-regulated, while that of E-cadherin down-regulated, in LNCaP/HIF1alpha. CONCLUSION: The over-expression of HIF-1alpha could induce EMT in the human prostate carcinoma cell line LNCaP and enhance its invasiveness through E-cadherin and vimentin regulation.

[Differential analysis of two-dimensional gel electrophoresis profiles of spermatozoal protein in human normal semen and idiopathic asthenospermia].

OBJECTIVES: To evaluate the application of two-dimensional electrophoresis in the research of differentially expressed proteins in human spermatozoa. METHODS: Two-dimensional gel electrophoresis was performed on 4 sperm samples from normal healthy men and another 4 from asthenospermia patients. After silver staining, the differential expression proteins were analyzed by PDQuest 2D analysis software. RESULTS: Seven differential protein spots were identified, 2 expressed highly in the asthenospermia sperm but lowly in the normal spermatozoa, while the other 5 expressed just the opposite way. CONCLUSION: The protein profiles of differential expression between the normal spermatozoa and idiopathic asthenospermia were established and some differential proteins were found. The data obtained in this study may help prepare the ground for further studies on the isolation and identification of differentially expressed proteins in human asthenospermia sperm.

[Over-expression of Hypoxia-inducible factor 1alpha increases invasive potency of LNCaP cells in vitro].

OBJECTIVE: To evaluate the effect of hypoxia-inducible factor-1alpha (HIF1alpha) over-expression on the invasive potency of human prostate cancer cell. METHODS: Human prostate cancer cells of the line LNCaP were cultured and transfected by the recombinant plasmid pcDNA3.1(-)-HIF-1alpha containing the gene HIF1alpha with Lipofectamine 2000 system. The positive clone cells were selected by G418 and confirmed by Western blotting and immunofluorescence staining (LNCaP/HIF1alpha cells). Transwell chambers with polycarbonate filter were coated by 100 microl Matrigel at 1:20 dilution in serum-free medium. LNCaP cell suspension and LNCaP/HIF1alpha cell suspension were inoculated into the Transwell chambers respectively for 24 hours to analyze the invasive potency. Western blotting was used to detect the expression of E-cadherin, vimentin, matrix metalloproteinase-2 (MMP-2), cathepsin D, and urokinase-type plasminogen activator receptor (uPAR). RESULTS: The expression level of HIF1alpha in the LNCaP/HIF1alpha cells was distinctly higher than that in the LNCaP cells. The numbers of LNCaP-HIF1alpha cells penetrating through the Transwell polycarbonate filter was 4.6 +/- 0.4 x 10(4), significantly higher than that of the LNCaP cells (3.2 +/- 0.3 x 10(4), P < 0.01). The expressions of vimentin, MMP-2, cathepsin D, and uPAR were all up-regulated in LNCaP-HIF1alpha cells than those of the LNCaP cells. Whereas, the expression of E-cadherin was down-regulated in the LNCaP-HIF1alpha cells. CONCLUSION: Over-expression of HIF-1alpha stimulates the invasion potency of human prostate carcinoma cell. The expression of E-cadherin, vimentin, MMP-2, cathepsin D, and uPAR, all playing an established role in the invasion of tumor, can be regulated by HIF-1 in human prostate cancer cell.

[Clinical trials of combined therapy of an oral Chinese medicine with massage for chronic nonbacterial prostatitis].

OBJECTIVE: To evaluate the clinical effect of an oral Chinese medicine combined with massage of the prostate in treating chronic nonbacterial prostatitis (CNP). METHODS: Seventy-two CNP patients were randomly divided into an experimental group (oral Chinese medicine with massage) and a control group (oral Chinese medicine only). The main parameters were compared before and two months after the treatment. RESULTS: Total efficacy rates of the experimental and control groups were 90.0% and 68.6%, respectively. Compared with the controls, patients in the experiment group had a lower score on NIH-CPSI, and the difference was significant statistically (P < 0.05). CONCLUSION: The combined therapy is more effective than oral Chinese medicine alone for the treatment of CNP.

Over-expression of hypoxia-inducible factor-1alpha increases the invasive potency of LNCaP cells in vitro.

OBJECTIVE: To evaluate the effect of hypoxia-inducible factor-1alpha (HIF-1alpha) over-expression on the invasion-associated proteins in human prostate cancer cells, as HIF-1alpha is a transcriptional factor that could activate genes involved in the response to hypoxia, but might also enhance the invasive potency of prostate cancer cells. MATERIALS AND METHODS: Prostate cancer (LNCaP) cells were transfected with recombinant plasmid pcDNA3.1(-)/HIF-1alpha and pcDNA3.1(-) control vector using a commercial system, designated as LNCaP/HIF-1alpha and LNCaP/pcDNA3.1, respectively. The positive clones were selected with G418 and confirmed by Western blot and indirect immunofluorescence labelling. A polycarbonate filter, coated with a matrix gel, was used to analyse the invasive potency. The expression of E-cadherin, vimentin, cathepsin D, matrix metalloproteinase (MMP2) and urokinase plasminogen activator receptor (uPAR) was detected by Western blot. RESULTS: The expression level of HIF-1alpha in LNCaP/HIF-1alpha was distinctly higher than that in LNCaP/pcDNA3.1 and LNCaP. Many more cells of LNCaP/HIF-1alpha penetrated through the polycarbonate filter than those of LNCaP/pcDNA3.1 and LNCaP. Compared with the LNCaP/pcDNA3.1 and LNCaP cells, the expression of vimentin, cathepsin D, MMP-2 and uPAR were up-regulated in LNCaP/HIF-1alpha, whereas the expression of E-cadherin was down-regulated. CONCLUSION: These results show that over-expression of HIF-1alpha directly stimulates the invasive potency of human prostate carcinoma LNCaP cells through the matrix gel. The expression of E-cadherin, vimentin, cathepsin D, MMP-2 and uPAR, which are proteins with established roles in the pathophysiology of invasion, could be regulated by HIF-1alpha in human prostate cancer cells.

Hypoxia-inducible factor-1alpha induces the epithelial-mesenchymal transition of human prostatecancer cells.

BACKGROUND: Hypoxia-inducible factor-1alpha (HIF-1alpha) is a transcriptional factor that could improve the stimulation of angiogenesis and the metabolic adaptation of tumor cells to hypoxia. A recent study showed that HIF-1alpha could induce colon cancer cells epithelial-mesenchymal transition (EMT). However, no evidence indicates a similar correlation in human prostate cancer cells. This study was designed to evaluate the effect of HIF-1alpha over-expression on the EMT in human prostate cancer cells. METHODS: We selected the appropriate cell line for HIF-1alpha induction from those EMT negative prostate cell lines through vimentin gene detection by RT-PCR. As the result, LNCaP cell line is the best one for further experiment. LNCaP cells were transfected with recombinant plasmid pcDNA3.1 (-)/HIF-1alpha and pcDNA3.1 (-) control vector by Lipofectamine 2000 system. The positive cell colonies were confirmed by indirect immunofluorescence labeling. Then Transwell polycarbonate filter was used to analyze the invasive potency. The expression of EMT associated proteins, E-cadherin and vimentin, was detected by Western blotting. RESULTS: Among four of the EMT negative cell lines, LNCaP was the only one expressed the vimentin gene but not the associated protein. The expression level of HIF-1alpha in LNCaP/HIF-1alpha was distinctly higher than that in LNCaP/pcDNA3.1 and LNCaP. The cell numbers of LNCaP/HIF-1alpha that penetrated through the Transwell filter were higher than that of LNCaP/pcDNA3.1 and LNCaP. Compared with the LNCaP/pcDNA3.1 and LNCaP cells, the expression of vimentin was up-regulated in LNCaP/HIF-1alpha, whereas the expression of E-cadherin was down-regulated. CONCLUSIONS: Over-expression of HIF-1alpha stimulates the invasion potency of human prostate carcinoma cells through EMT pathway. The expression of E-cadherin and vimentin, playing established roles in EMT, could be regulated by HIF-1alpha in human prostate cancer cell line.

[Expression and significance of metastasis-associated proteins in prostate cancer cell lines with different metastatic potentials].

OBJECTIVE: To observe the expression profiles between two metastasis-associated proteins in different prostate cancer cell lines and explore the molecular mechanisms of bone metastatic potentials. METHODS: Expressions of E-cadherin and vimentin in two prostate cancer cell lines (LNCaP and IA8) with different metastatic potentials were detected by Western blotting. RESULTS: There was remarkable difference in the expressions of E-cadherin and vimentin between the highly metastatic cell line and the lowly one. As one of the adhesion associated proteins, E-cadherin was detected with high level of expression in LNCaP cell line, which was well known as low metastatic potential. However, E-cadherin did not expressed in IA8 with high metastatic potential. And as one of the cytoskeleton proteins, vimentin expression was high in IA8, but not in LNCaP. CONCLUSION: There is definitely difference in the metastatic phenotypes (E-cadherin and vimentin) among cell lines with different metastatic potentials. The expressions of E-cadherin and vimentin proteins may play important roles in promoting and inhibiting the metastasis of prostate cancer respectively, and may be considered to be valuable in evaluating the malignant degree, predictable metastasis and prognosis of prostate cancers.