Phei-Lang Chang

Xanthogranulomatous Pyelonephritis: Experience in 36 Cases

A retrospective study of 36 patients with xanthogranulomatous pyelonephritis who underwent nephrectomy at our hospital was performed. The disease occurred most frequently in middle-aged women with a history of recurrent urinary tract disorder. There were 2 cases of focal xanthogranulomatous pyelonephritis, 2 associated with emphysematous pyelonephritis, 2 that manifested as fistula formation between the colon or skin, and 1 with deep sinus formation into the hip joint that presented as septic arthritis. Flank pain and fever were the most frequent complaints. Escherichia coli (67%) and Proteus mirabilis (26%) were the most common organisms isolated from the voided urine, kidney and blood stream. Cephalothin plus gentamicin or tobramycin were the drugs of choice before surgical intervention.

Comparative and Targeted Proteomic Analyses of Urinary Microparticles from Bladder Cancer and Hernia Patients

Bladder cancer is a common urologic cancer whose incidence continues to rise annually. Urinary microparticles are an attractive material for noninvasive bladder cancer biomarker discovery. In this study, we applied isotopic dimethylation labeling coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) to discover bladder cancer biomarkers in urinary microparticles isolated from hernia (control) and bladder cancer patients. This approach identified 2964 proteins based on more than two distinct peptides, of which 2058 had not previously been reported as constituents of human urine exosomes/microparticles. A total of 107 differentially expressed proteins were identified as candidate biomarkers. Differences in the concentrations of 29 proteins (41 signature peptides) were precisely quantified by LC-MRM/MS in 48 urine samples of bladder cancer, hernia, and urinary tract infection/hematuria. Concentrations of 24 proteins changed significantly (p<0.05) between bladder cancer (n=28) and hernia (n=12), with area-under-the-curve values ranging from 0.702 to 0.896. Finally, we quantified tumor-associated calcium-signal transducer 2 (TACSTD2) in raw urine specimens (n=221) using a commercial ELISA and confirmed its potential value for diagnosis of bladder cancer. Our study reveals a strong association of TACSTD2 with bladder cancer and highlights the potential of human urinary microparticles in the noninvasive diagnosis of bladder cancer.

Discovery of novel bladder cancer biomarkers by comparative urine proteomics using iTRAQ technology.

A urine sample preparation workflow for the iTRAQ (isobaric tag for relative and absolute quantitation) technique was established. The reproducibility of this platform was evaluated and applied to discover proteins with differential levels between pooled urine samples from nontumor controls and three bladder cancer patient subgroups with different grades/stages (a total of 14 controls and 23 cancer cases in two multiplex iTRAQ runs). Combining the results of two independent clinical sample sets, a total of 638 urine proteins were identified. Among them, 55 proteins consistently showed >2-fold differences in both sample sets. Western blot analyses of individual urine samples confirmed that the levels of apolipoprotein A-I (APOA1), apolipoprotein A-II, heparin cofactor 2 precursor and peroxiredoxin-2 were significantly elevated in bladder cancer urine specimens (n = 25-74). Finally, we quantified APOA1 in a number of urine samples using a commercial ELISA and confirmed again its potential value for diagnosis (n = 126, 94.6% sensitivity and 92.0% specificity at a cutoff value of 11.16 ng/mL) and early detection (n = 71, 83.8% sensitivity and 94.0% specificity). Collectively, our results provide the first iTRAQ-based quantitative profile of bladder cancer urine proteins and represent a valuable resource for the discovery of bladder cancer markers.

Triiodothyronine Modulates Cell Proliferation of Human Prostatic Carcinoma Cells by Downregulation of the B-Cell Translocation Gene 2

Studies suggest that triiodothyronine (T3) and cognate nuclear receptors (hTR) are involved in regulation of prostatic cell growth and differentiation. To probe mechanisms for T3 effects, we studied prostate carcinoma cells, investigating the effect of T3 on expression of the B‐cell translocation gene 2 (BTG2), which regulates the G1/S transition of the cell cycle.

Curcumin Blocks the Activation of Androgen and Interlukin-6 on Prostate-Specific Antigen Expression in Human Prostatic Carcinoma Cells

Curcumin, a naturally occurring compound, exhibits anticancer chemopreventive effects. We evaluated the effects and mechanisms of curcumin on the gene expression of prostate-specific antigen (PSA) in human androgen-sensitive prostatic carcinoma cells. LNCaP cells were used to determine the effect of curcumin on PSA expression. Quantitative PSA expression was assessed by reverse transcription polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA), and immunoblot assay. The modulation of androgen, interlukin-6 (IL-6), and prostate-derived Ets factor (PDEF) on the PSA gene was identified by transient gene expression assay with the use of a PSA reporter vector. The effect of curcumin on the activity of androgen receptors was evaluated by electrophoretic mobility shift assay (EMSA). Immunoblot assays, RT-PCR, and ELISA indicated that curcumin treatments blocked the stimulation of methyltrienolone (R1881) and IL-6 on PSA gene expression in LNCaP cells. The effects of curcumin appear to be mediated via the androgen response element of PSA gene. Results from immunoblot assay and EMSA revealed the modulation of curcumin on the expression of androgen receptor and androgen receptor binding activity on androgen response element of PSA gene. Although overexpression of PDEF dramatically enhanced PSA gene expression, the results of immunoblot assays and transient reporter assays indicated that curcumin treatments did not affect the gene expression of PDEF. Curcumin inhibits R1881- and IL-6-mediated PSA gene expression in LNCaP cells through down-regulation of the expression and activity of androgen receptors.

l-Mimosine blocks cell proliferation via upregulation of B-cell translocation gene 2 and N-myc downstream regulated gene 1 in prostate carcinoma cells

L-Mimosine, an iron chelator and a prolyl 4-hydroxylase inhibitor, blocks many cancer cells at the late G1 phase. B-cell translocation gene 2 (Btg2) regulates the G1/S transition phases of the cell cycle. N-myc downstream regulated gene 1 (Ndrg1) is a differentiation-inducing gene upregulated by hypoxia. We evaluated the molecular mechanisms of L-mimosine on cell cycle modulation in PC-3 and LNCaP prostate carcinoma cells. The effect of L-mimosine on cell proliferation of prostate carcinoma cells was determined by the [3H]thymidine incorporation and flow cytometry assays. L-Mimosine arrested the cell cycle at the G1 phase in PC-3 cells and at the S phase in LNCaP cells, thus attenuating cell proliferation. Immunoblot assays indicated that hypoxia and L-mimosine stabilized hypoxia-inducible factor-1α (HIF-1α) and induced Btg2 and Ndrg1 protein expression, but downregulated protein levels of cyclin A in both PC-3 and LNCaP cells. L-Mimosine treatment decreased cyclin D1 protein in PC-3 cells, but not in LNCaP cells. Dimethyloxalylglycine, a pan-prolyl hydroxylase inhibitor, also induced Btg2 and Ndrg1 protein expression in LNCaP cells. The transient gene expression assay revealed that L-mimosine treatment or cotransfection with HIF-1α expression vector enhanced the promoter activities of Btg2 and Ndrg1 genes. Knockdown of HIF-1α attenuated the increasing protein levels of both Btg2 and Ndrg1 by hypoxia or L-mimosine in LNCaP cells. Our results indicated that hypoxia and L-mimosine modulated Btg2 and Ndrg1 at the transcriptional level, which is dependent on HIF-1α. L-Mimosine enhanced expression of Btg2 and Ndrg1, which attenuated cell proliferation of the PC-3 and LNCaP prostate carcinoma cells.

Upregulation of prostate-derived Ets factor by luteolin causes inhibition of cell proliferation and cell invasion in prostate carcinoma cells

Luteolin is a polyphenolic flavone and has antitumor activity for many cancers. The prostate‐derived Ets factor (PDEF), a novel epithelium‐specific Ets transcription factor, acts as an androgen‐independent transcriptional activator of the prostate‐specific antigen (PSA) promoter. We determined the antitumor function of luteolin via upregulation of PDEF gene expression in human prostate carcinoma LNCaP cells. Results from flow cytometry and 3H‐thymidine incorporation assays revealed that luteolin treatments attenuated cell proliferation and arrested the cell cycle at the G1/S phase. High concentration of luteolin (30 μM) induced cell apoptosis. Immunoblot assays and enzyme linked immunosorbent assay revealed that luteolin treatment upregulated PDEF but downregulated androgen receptor (AR) gene expression, which decreased PSA gene expression in LNCaP cells. Results of immunoblot and transient gene expression assays revealed that luteolin treatments at proapoptosis dosage, enhanced gene expression of PDEF, B‐cell translocation gene 2 (BTG2), N‐myc downstream regulated gene 1 (NDRG1) and Maspin. Transient gene expression assays indicated that cotransfection of the PDEF expression vector enhanced the promoter activities of the BTG2, NDRG1 and Maspin genes. Stable overexpression of PDEF significantly induced BTG2, NDRG1 and Maspin gene expression, which markedly attenuated in vitro cell proliferation and invasion of LNCaP cells. The modulatory effect of luteolin on BTG2, NDRG1 and Maspin gene expression were attenuated when PDEF was knocked‐down. These results suggest that luteolin blocks PSA gene expression by downregulation of AR expression. The enhancement of PDEF expression, which induced BTG2, NDRG1 and Maspin gene expression, could account for the function of luteolin for antiproliferation and anti‐invasion in LNCaP cells.

p53 Downregulates the Gene Expression of Mitochondrial Aconitase in Human Prostate Carcinoma Cells

Mitochondrial aconitase (mACON) is regarded as the key enzyme in citrate oxidation in human prostate epithelial cells, and its abnormal expression has been implicated in tumorigenesis of the prostate. Evidence also supports a broad role for the p53 gene in suppressing prostatic tumorigenesis. We investigated whether p53 regulates mACON expression and explore the potential mechanisms responsible for its effect on prostate cancer cells.

Association of nucleophosmin/B23 with bladder cancer recurrence based on immunohistochemical assessment in clinical samples.

AbstractAim:To investigate the possible correlation of nucleophosmin/B23 expression with bladder carcinoma recurrence.Methods:Surgically-resected bladder tumors staged pTa to pT4 were examined for nucleophosmin/B23 expression by immuno-histochemistry. The study group consisted of 132 consecutive patients surgically treated at Chang Gung Memorial Hospital between December 1998 and November 1999. The mean follow up was 72 months (range: 48-84 months).Results:Nuclear nucleophosmin/B23 staining was detected in 96% of advanced stage and poorly-differentiated tumors. Higher nucleophosmin/B23 levels were linked to more advanced tumor stages, grades, poor prognosis, and likelihood of recurrence (P<0.05). The Cox multivariate analysis indicated the nucleophosmin/B23 expression as an independent indicator for tumor recurrence (P=0.009).Conclusion:The results suggest that nucleophosmin/B23 is a favorable prognostic indicator for bladder cancer. Nucleophosmin/B23 could be a useful molecular tumor marker for predicting bladder cancer recurrence.

A therapeutic trial of acupuncture in neurogenic bladder of spinal cord injured patients : a preliminary report

Eighty patients with spinal cord injury (SCI) and neurogenic bladder were studied. Among them, 28 (70%) cases in the control group and 32 (80%) cases in the electroacupuncture group achieved ultimately balanced voiding and were selected for further analysis. The acupuncture group received electroacupuncture at four acupoints: Chung Chi (Conception Vessel CV3), Kuan Yuan (CV4), and bilateral Tzu Liao (Urinary Bladder UB32), in addition to conventional intermittent catheterization program (ICP). Whereas the control group underwent conventional bladder training program with ICP only. The results revealed that the time of achieving balanced voiding was statistically significantly shorter with electroacupuncture than in the control group: 57.1±22.5 vs 85.2±27.4 days (P<0.005) for upper motor neuron lesions, and 55.4±22.6 vs 83.4±26.1 days (P<0.01) for lower motor neuron lesions. However, there was almost no difference between upper motor neuron lesions and lower motor neuron lesions. When acupuncture commenced was also a factor to influence the results. Those who received acupuncture within 3 weeks after injury had significantly shortened the total days to achieve a balanced bladder, as compared to those who received acupuncture 3 weeks after injury (46.6±13.2 vs 65.8±15.4 days, P<0.005). Our study implied that acupuncture might be beneficial in the management of neurogenic bladder of SCI, and the earlier the patient received electro-acupuncture therapy, the sooner the bladder balanced. On the other hand, we also found that complete spinal cord injury, either with pronounced detrusor-sphincter dyssynergia in upper motor nueron lesion or with persistent areflexic bladder in lower motor neuron lesion, was not affected by acupuncture.