PBOV1 promotes prostate cancer proliferation by promoting G1/S transition.

Prostate cancer (PC) is one of the leading causes of cancer death in men, and thus, finding new regulators is critical for PC therapy. Prostate and breast cancer overexpressed 1 (PBOV1) is overexpressed in breast, prostate, and bladder cancers, as it is upregulated in the serum of patients with PC, but the role of PBOV1 in PC has not been studied. In this article, we found that PBOV1 was indeed overexpressed in PC cells; PBOV1 overexpression promoted cell proliferation and colony formation ability and arrested cell cycle in the G0/G1 phase and tumorigenicity ability in vitro, whereas knockdown of PBOV1 reduced these effects. Further analysis of PBOV1 overexpression inhibited cell cycle inhibitors, P21 and P27, and increased the phosphorylation level of Rb and cyclin D1 expression, suggesting that PBOV1 promoted cell proliferation through promoting G1/S transition.

Flank-suspended versus prone percutaneous nephrolithotomy: changes of haemodynamics, arterial blood gases and subjective feelings.

BACKGROUND: The flank-suspended position was adopted in percutaneous nephrolithotomy (PCNL), and haemodynamics, blood gas variables and subjective feelings were examined with an attempt to explore the effect of the operative position in PCNL on the body's inner environment and patient comfort. OBJECTIVE: The influence of the flank-suspended and prone position on haemodynamics, arterial blood gases and subjective feelings in patients receiving PCNL was examined. DESIGN, SETTING AND PARTICIPANTS: A total of 100 patients with kidney stones who underwent PCNL during January 2010 to January 2011 were divided into flank-suspended groups (n = 50) and prone groups (n = 50) at random in terms of the operative position. The blood pressure, heart rate, respiratory frequency and oxyhaemoglobin saturation and blood gas variables were determined at different time points (before the operation, after position change, 30 min after the start the operation and immediately after the operation). Visual analogue scale (VAS) scoring system was employed to define the posture comfort, dyspnoea and puncture-site pain 24 h postoperation in the patients. All the measures were compared between patients in different positions at different time points. STATISTICAL ANALYSIS: Paired t-test was employed in the comparison of measures detected at different time points in the same group. Two-group comparison was subjected to t-test. A p value less than 0.05 was considered statistically significant. RESULTS AND LIMITATIONS: The blood pressure was decreased within and after the operation in both groups, substantially in the prone group, significantly lower than that before the operation (p<0.05). No significant differences in the heart rate, respiratory frequency and oxyhaemoglobin saturation were noted among the different time points in the same group. Blood gas analysis showed that pH value and base excess were profoundly reduced within and after the operation in the two groups, significantly lower than those before the operation, and the decrease was most manifest in the prone group. There was no difference in the blood sodium and potassium among the different time points in each group. The flank-suspended group was superior to the prone group with regard to posture comfort and dyspnoea degree but not puncture-site pain 24 h postoperation. CONCLUSIONS: Flank-suspended and prone PCNL affects the haemodynamics, blood gas variables and subjective feelings of patients to a varying degree. The flank-suspended PCNL possesses advantages over prone PCNL such as little influence on haemodynamics and blood gas variables, satisfactory posture comfort, less dyspnoea and easy access to vital sign observation.

Elevated expression of glutaminase confers glucose utilization via glutaminolysis in prostate cancer.

Cancer cells reprogram their metabolism towards aerobic glycolysis and elevated glutaminolysis, which contributes to the aggressive phenotype. Understanding how these metabolic pathways are regulated may provide critical targets for therapeutic intervention. Glutaminase (GLS1) is a key enzyme that converts glutamine to glutamate. In this study, we show the loss of GLS1 function by RNA interference or inhibitor diminished the rates of glucose utilization, growth and invasiveness of prostate cancer cells. We propose that GLS1 positively regulates glucose uptake in addition to glutaminolysis. Further, GLS1 involves the transcriptional repression of thioredoxin interacting protein (TXNIP), which is a potent negative regulator of glucose uptake and aerobic glycolysis. Most importantly, we provided direct evidence that elevated GLS1 expression was highly correlated with the tumor stage and progression in prostate cancer patients. Together, we defined a key role for GLS1 in coupling glutaminolysis of the TCA cycle with elevated glucose uptake and consequently the growth of prostate cancer cells. These data extends the role of GLS1 in regulating cell metabolism and the clinical utility of GLS1 inhibitors in the restriction of essential nutrients.

[Effects of adenovirus-mediated PTEN on the proliferation of prostate cancer PC-3 cells and expressions of cyclin D1 and p21].

OBJECTIVE: To construct a recombinant adenovirus expression vector containing the anti-oncogene PTEN and to investigate the effects of the PTEN gene on the proliferation of prostate cancer PC-3 cells and the expressions of cyclin D1 and p21 in the PC-3 cells. METHODS: The PTEN gene was amplified from the rat hippocampus by RT-PCR and cloned into the shuttle plasmid pEN-TR2A. The plasmids were constructed and amplified in 293A cells. Prostate cancer PC-3 cells were cultured in vitro and infected with the adenoviral vector carrying the PTEN gene (Ad-PTEN). The up-regulation of the PTEN protein was measured by indirect immuno-fluorescence assay; the expressions of PTEN, cyclin D1 and p21 in the cells infected with Ad-PTEN and Ad-LacZ were determined by RESULTS: The Western blot; and the effect of PTEN on the cell proliferation was detected by MTT assay and plate colony formation. recombinant adenoviral vector Ad-PTEN was successfully constructed. Western blot showed a significantly increased expression of the PTEN protein in the PC-3 cells infected with Ad-PTIEN (0.215 +/-0.065) as compared with that in the control ([0.052 +/-0.009], t = 4. 30, P <0.05) and the Ad-LacZ group ( [0. 056 +/- 0.008 ] , t =4.21, P <0.05). The expression of cyclin D1 was significantly lower in the Ad-PTEN-infected PC-3 cells (0. 256 +/- 0. 072) than in the control ( [0. 502 +/- 0. 087 ], t = 3.77, P < 0.05) and the Ad-LacZ group ([0.498 +/-0.081] , t =3.87, P <0.05), while the expression of p21 remarkably higher in the Ad-PTEN-infected PC-3 cells (0.589 +/-0. 076) than in the control ([0. 146 +/-0.026] , t = 9.55, P<0. 01) and the Ad-LacZ group ([0. 163 +/-0. 024] , t = 9.26, P <0.01). Ad-PTEN significantly inhibited the growth of the PC-3 cells (21.98%) at 48 h (t = 6.80, P <0.01). The colony formation rate of the PC-3 cells was (37.4 +/-4. 18)% in the Ad-PTEN group, significantly lower than (54.9 +/-4.81)% in the control (t =4.76, P<0.01) and (56.5 +/- 5.42)% in the Ad-LacZ group (t=4.83, P<0.01). CONCLUSION: The expression of PTEN induced by Ad-PTEN can significantly inhibit the proliferation of PC-3 cells, down-regulate the expression of cyclin D1, and up-regulate the expression of p21.

Upstream and downstream mechanisms for the promoting effects of IGF-1 on differentiation of spermatogonia to primary spermatocytes.

AIM: The aim of this study is to delineate the mechanisms for the promoting the effects of insulin growth factor I (IGF1) on the differentiation of spermatogonia into primary spermatocytes. MAIN METHODS: We used organ culture of testicular fragments from mice to observe the effects of varying agents and siRNAs. Real-time RT-PCR and immunoblotting analysis were employed to quantify expression of genes. Luciferase reporter gene assay was employed for verifying the targeting relationship between miRNA and protein-coding genes. KEY FINDINGS: During spermatogenesis while spermatozoa pass through epididymis and vas deferens for maturation, expression of IGF1 and its receptor IGF1R, p44/ERK1 and p42/ERK2, and PI3K was all upregulated, whereas let-7 miRNA family members let-7a/b/d/e/g/ were downregulated. We established both IGF1 and IGF1R as cognate target genes for let-7; downregulation of let-7 resulted in upregulation of IGF1 and IGF1R during the early stage of differentiation from spermatogonia to primary spermatocytes. Transfection of let-7 inhibited, whereas transfection of anti-let-7 inhibitor enhanced the differentiation. The promoting effect of anti-let-7 was eliminated by PPP to block IGF1R phosphorylation. IGF1 activated ERK1/2 and PI3K and induced differentiation. PPP eliminated the activation of ERK1/2 and PI3K and inhibited the differentiation induced by IGF1. Specific inhibition of ERK1/2 by U0126 or PI3K by LY294002 reduced the IGF1-induced differentiation. Knockdown of ERK1/ERK2 or PI3K by siRNAs also blocked IGF1-induced spermatogenesis. SIGNIFICANCE: Our study therefore identified downregulation of let-7 as an upstream mechanism for IGF1/IGF1R upregulation and activation of ERK1/2 and PI3K as a downstream mechanism mediating the IGF1 signaling cascade promoting differentiation of spermatogonia to primary spermatocytes.

Flank suspended supine position for percutaneous nephrolithotomy.

AIMS: Prone and supine positions for percutaneous nephrolithotomy are widely used but have their drawbacks. We report a new positioning method called "flank suspended supine position" (FSSP) for PCNL and describe our experience with PCNL in this position to evaluate its safety and efficacy. METHODS: Retrospective study of 150 cases of renal stone patients treated with PCNL in a new position called flank suspended supine position (FSSP) from June 2009 to July 2010. All patients were treated with PCNL in FSSP under epidural anesthesia. Operation time, bleeding rate, stone free rate, and complications were recorded. RESULTS: All patients tolerated FSSP. Mean operation time was 78.29±26.13 min. Initial stone-free rate was 83%. For those with residual stones (26 cases), 18 were stone-free after a second PCNL, 8 after extracorporeal shock wave lithotripsy (ESWL). Mean hospital stay was 7.63±2.39 days. No penetrating injury of the pleural cavity or injury to visceral organs was reported. SUMMARY: FSSP is an effective and safe position for PCNL in our hands and its effectiveness relative to traditional prone position needs to be determined in future randomized studies.

[The changes of IGF-I in testis and epididymis on a rat model with oligozoospermia/azoospermia induced by cyclophosphamide].

OBJECTIVE: To evaluate the effect of the levels of IGF-I in the epididymis and the expression of IGF-I in the testis of adult male rat after the administration of cyclophosphamide. METHODS: Ninety-six male adult rats (8 weeks age) were divided into 6 groups. The doses given to the rats of the groups 1 to 5 were 10, 20, 40, 80 and 100 mg/(kg x d), respectively. The remaining group was served as control. All those rats were sacrificed and IGF-I were quantitatively determined by ELISA techniques 2 and 4 weeks after the administration of the drug (by gastric fudge). Immunohistochemical SP technique was used to examine expression of IGF-I in rat testis. RESULTS: The levels of cell factors (IGF-I) in the epididymis of the rats were gradually reduced with the increasing time and dose after administration of the drug. In the mean time the expression of IGF-I in the tissues of the testis of those rats were also gradually reduced. CONCLUSION: In the time of oligozoospermia/azoospermia induced by the administration of cyclophosphamide, the expression levels of IGF-I in the genetic system were significantly reduced. The possible mechanism of these changes could be attributed to the lower spermatogenesis function of the testis caused by the administration of cyclophosphamide.

Elevated autoantibodies against oxidized palmitoyl arachidonoyl phosphocholine in patients with hypertension and myocardial infarction.

Oxidized 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine (oxPAPC) is antigenic and an important epitope of oxLDL. This study validates the assay for autoantibodies against oxPAPC (anti-oxPAPC-Ab) and investigates the possible association between anti-oxPAPC-Ab and cardiovascular disease. A synthetic PAPC was oxidatively modified as an antigen for the anti-oxPAPC-Ab assay. The concentrations of the antibody in serum were measured by EIA. The analytical parameters of the anti-oxPAPC-Ab assay were validated. Levels of anti-oxPAPC-Ab were prevalent in patients with hypertension, myocardial infarction (MI) and healthy subjects. Anti-oxPAPC-Ab specifically reacts with oxPAPC, but not with 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine (PAPC). The characteristics of the assay included precision (inter-assay coefficients of variation were 7.9% for IgG and 13.2% for IgM), cross-reactivity, clinical sensitivity for hypertension (43% and 47%) and MI (37% and 41%), clinical specificity (95.2%) and normal values (less than 13 Unit/mL for IgG and less than 7 Unit/mL for IgM). Elevated levels of anti-oxPAPC-Ab were found in smoking populations, in patients with hypertension and MI. Anti-oxPAPC-Ab are significantly elevated in patients with hypertension and MI. A synthetic PAPC, after oxidation, was used to detect anti-oxPAPC-Ab, which may greatly enhance the reliability of the assay. The determination of anti-oxPAPC-Ab could serve as an autoimmune marker in the associating diagnosis of cardiovascular disease.

Significance of the detection of HIV-1 gag- and/or pol-CD8/A2 T-lymphocytes in HIV-patients.

UNLABELLED: Cytotoxic T-lymphocytes (CTL) play an important role in the immune system's defense against human immunodeficiency virus (HIV) infection. The functional status of CTL closely relates to the progression of HIV disease. We have validated the characteristics of the assay for HIV-1 gag- and pol-specific-CD8/HLA-A2 T-cells from peripheral blood by flow cytometry. Sixty-nine healthy individuals and 38 HIV-patients with HLA-A2 antigen-positive subjects were included in the study. Neither HIV-1 gag- nor pol-specific-CD8/HLA-A2 T-cells were determined in these healthy subjects. HIV-1 gag- and pol-specific-CD8/HLA-A2 T-cells could be detected in HIV-patients. The frequency of specific CTL was 58% (22/38) in the patient group. There was a significantly inverse correlation (p < 0.05) between HIV-1 gag- and pol-specific-CD8/HLA-A2 T-cells and HIV plasma viremia in the patients. CONCLUSION: The HIV-1 gag- or pol-specific-CD8/HLA-A2 T-cells assay is sensitive and specific, being able to detect at the single T-cell level. This assay may provide a versatile tool for structured HIV treatment and for monitoring vaccination efficacy.