ROS-NLRP3 signaling pathway induces sterile inflammation after thulium laser resection of the prostate.

The sterile inflammation (SI) of the urinary tract is a common problem requiring serious consideration after prostatectomy. This study mainly focuses on the role of the reactive oxygen species-NLR family, pyrin domain-containing 3 (ROS-NLRP3) signaling pathway in SI after thulium laser resection of the prostate (TmLRP). Urinary cytokines were determined in patients who received TmLRP, and heat shock protein 70 (HSP70) was detected in the resected tissues. The involvement of ROS signaling in HSP70-induced inflammation was explored in THP-1 cells with or without N-acetyl- l-cysteine (NAC) pretreatment. The function of NLRP3 and Caspase-1 was determined by Western blot analysis, enzyme-linked immunosorbent assay (ELISA), and polymerase chain reaction. These phenomena and mechanisms were verified by the beagle models that received TmLRP. Clinical urine samples after TmLRP showed high expression of inflammatory factors and peaked 3-5 days after surgery. The high expression of HSP70 in the resected tissues was observed. After HSP70 stimulation, the expression of ROS, NLRP3, Caspase-1, and interleukin-18 (IL-18) increased significantly and could be reduced by ROS inhibitor NAC. The expression of IL-1ß and IL-18 could be inhibited by NLRP3 or Caspase-1 inhibitors. In beagle models that received TmLRP, HSP70, NLRP3, Caspase-1, IL-1ß, and IL-18 were highly expressed in the wound tissue or urine, and could also be reduced by NAC pretreatment. Activation of the ROS-NLRP3 signaling pathway induces SI in the wound after prostatectomy. Inhibition of this pathway may be effective for clinical prevention and treatment of SI and related complications after prostatectomy.

Long non-coding RNA lnc-MX1-1 is associated with poor clinical features and promotes cellular proliferation and invasiveness in prostate cancer.

Long non-coding RNAs (lncRNAs) are emerging as key molecules in human cancer genesis and progression, including prostate cancer. Large amount of lncRNAs have been found that differentially expressed between prostate cancer tissues and normal prostate tissues. Whether these lncRNAs could serve as a novel biomarker for prostate cancer diagnosis or prognosis, and their biological functions in prostate cancer need further investigation. In the present study, we identified that lncRNA lnc-MX1-1 is over-expressed in prostate cancer tissues compared with their adjacent normal prostate tissues by gene expression array profiling. The expression of lnc-MX1-1 in 60 prostate cancer cases was determined by real-time quantitative PCR and the correlations between lnc-MX1-1 expression and patients' clinical features were further analyzed. Next, we impaired lnc-MX1-1 expression using RNAi in LNCaP and 22Rv1 prostate cancer cells to explore the effects of lnc-MX1-1 on proliferation and invasiveness of the cells. Our results showed that there was a significant association between over-expression of lnc-MX1-1 and patients' clinical features such as PSA, Gleason score, metastasis, and recurrence free survival. Moreover, knockdown of lnc-MX1-1 reduced both proliferation and invasiveness of LNCaP and 22Rv1 cells. In conclusion, the results suggest that lnc-MX1-1 may serve as a potential biomarker and therapeutic target for prostate cancer.

Clinical evaluation and technical features of three-dimensional laparoscopic partial nephrectomy with selective segmental artery clamping.

PURPOSE: To investigate the feasibility and effectiveness of three-dimensional laparoscopic partial nephrectomy with selective segmental artery clamping (3D-LPNSSAC) comparing with the conventional two-dimensional laparoscopic partial nephrectomy (2D-LPN). METHODS: Between November 2012 and March 2014, 90 patients with cT1 renal tumor at Shanghai General Hospital were enrolled in our study, which were randomly divided into two groups: the 3D-LPNSSAC group (n = 45) and 2D-LPN group (n = 45). The perioperative variables, including operative time, dissecting time, suturing time, blood loss, warm ischemia time (WIT), preoperative and postoperative renal functions, were recorded and analyzed. In addition, the oncological outcomes and complications were also evaluated. RESULTS: All the LPNs were performed successfully without conversion to radical nephrectomy or open surgery, only three cases were converted to total renal artery clamping during 3D-LPNSSAC. There were no significant differences in operative time and dissecting time between the groups, while the suturing time was shorter during 3D-LPNSSAC (P < 0.01). The technique was associated with higher blood loss (P < 0.01). The technique of 3D-LPNSSAC significantly reduced WIT (P = 0.04), and better postoperative ipsilateral renal function could be obtained during 3D-LPNSSAC (P < 0.01). During a mean follow-up time of 16.8 months (range 5.5-22.5 months), the complication rate was 8.8 % (8/90) and no tumor reoccurrence was detected. CONCLUSIONS: 3D-LPNSSAC is a feasible and safe technique for treating selective renal tumors, presenting with the beneficial clinical outcomes of reduced suturing time, shorter WIT and better postoperative ipsilateral renal function.

The toxic effect of opioid analgesics on human sperm motility in vitro.

Opioid analgesics are the most common therapeutic analgesic for acute pain. In this study, the toxicological and pharmacological features of a group of opioid analgesics were characterized by the motility of human sperm. Aliquots of sperm were incubated with various concentrations of opioid analgesics in vitro. Computer-assisted sperm analysis was used to assess sperm motility at 15 minutes, 2 hours, and 4 hours after drug addition to the medium. Butorphanol and dezocine showed marked reduction of motility after incubation with sperm for 15 minutes. Butorphanol was more effective than dezocine in immobilizing sperm. Other opioids studied, such as fentanyl, alfentanil, and sufentanil, showed only partial inhibitory activity. Based on the data reported herein, we have found that butorphanol and dezocine exert a sperm-immobilizing effect. However, fentanyl, alfentanil, and sufentanil exhibit only partial inhibition of sperm motility. Given the increasing use of opioids and their potential effect on sperm motility, these findings are greatly relevant to male reproductive health.

[Effects of opioid analgesics on sperm motility in vitro].

OBJECTIVE: To observe the effects of short-term exposure to opioid analgesics on human sperm motility. METHODS: Twenty normal semen samples were collected, each divided into 19 groups, one as the control and the others treated in vitro with six opioid analgesics at three different concentrations, respectively, and sperm motility was assessed by computer-assisted sperm analysis at 15 min, 2 h and 4 h. RESULTS: Compared with the control group, fentanyl, alfentanil and sufentanil at 1 x 10(-5), 2 x 10(-3) and 0.05 mg/ml significantly decreased the percentage of grade a + b sperm at 15 min, 2 h and 4 h (P<0.05), and so did butorphanol at 2 x 10(-3) and 0.05 mg/ml (P<0.05) and dezocine at 0.05 and 0.5 mg/ml (P<0.05), but neither showed any remarkable effect at 1 x 10(-5) mg/ml at the three time points (P>0.05). Pentazocine effected no significant difference at 3 x 10(-5) and 0. 05 mg/ml (P>0.05) but a gradual increase in the percentage of grade a + b sperm at 0.5 mg/ml at the three time points (P<0.05). Butorphanol totally inhibited sperm motility at 0.05 mg/ml at 15 min and at 2 x 10(-3) mg/ml at 2 h, and so did dezocine at 0.05 and 0.5 mg/ml, but such inhibitory effect was not observed with fentanil, alfentanil and sulfentanil at 0.05 mg/ml. As for the sperm motility decreasing effect at 0.05 mg/ml at 15 min, sufentanil, butorphanol and dezocine exhibited significant differences (P<0.05) while fentanyl displayed none from alfentanil (P>0.05). CONCLUSION: Given the same length of time of treatment, butorphanol and dezocine totally inhibit sperm motility at a high concentration, but make no significant change at a low concentration. While fentanyl, alfentanil and sufentanil can significantly decrease sperm motility at the same low concentration, and partially inhibit it at all concentrations. On the contrary, a high concentration of pentazocine can promote human sperm motility.

The effect of repeated etomidate anesthesia on adrenocortical function during a course of electroconvulsive therapy.

BACKGROUND: Etomidate may affect adrenocortical function. We conducted an investigation of the comparative effects of etomidate and propofol during electroconvulsive therapy (ECT) on adrenocortical function and hemodynamics. METHODS: Patients in group T received etomidate and those in group B received propofol during intravenous anesthesia in ECT. Patients underwent ECT once every 2 days for 6 times. The serum levels of cortisol (Cor) and adrenocorticotropic hormone were determined 5 minutes before first anesthesia (baseline level, D0), and 24 hours (D1) as well as 48 hours after the last ECT (D2). At the same time, the hemodynamics was measured 2 minutes before anesthetic induction (T0), 30 seconds (T1) and 20 minutes after ECT (T2). Electrographic seizure duration (t), average seizure energy index, and postictal suppression index were recorded. RESULTS: Compared with the baseline level, serum Cor levels in group T were markedly decreased, but in normal ranges, at 24 hours after second and sixth treatments. No significant difference in serum Cor level was observed between the baseline and 48 hours posttreatment. In group B, there was no significant difference in serum Cor level between the baseline and 24 hours as well as 48 hours after each treatment. Furthermore, no significant difference in adrenocorticotropic hormone level was observed between the baseline and 24 hours as well as 48 hours posttreatment. However, the hemodynamics markedly changed during ECT and reached the preanesthetic level at 20 minutes posttreatment. The ECT-induced seizure duration in group T was longer than that in group B. However, seizure energy index and postictal suppression index was not significantly different between groups T and B. CONCLUSIONS: Etomidate and propofol would not affect the adrenocortical function during ECT, and hemodynamics reached normal level in a short time after ECT. Etomidate and propofol were both safe intravenous anesthetics during ECT, although etomidate was associated with comparatively longer seizure duration.

[Effects of remifentanil combined with naloxone on human sperm motility].

OBJECTIVE: To observe the effects of remifentanil combined with naloxone on human sperm motility in vitro and to investigate its possible mechanism. METHODS: Twenty normal semen samples were collected, each divided into 13 aliquots, one as the control and the others treated in vitro with different concentrations of remifentanil or remifentanil + naloxone for 35 min. The percentage of progressive mobile sperm was assessed by computer-assisted sperm analysis at 5, 10, 15, 20 and 35 min. RESULTS: Compared with the control group, remifentanil at 0.1, 1, 10 and 100 microg/L significantly decreased sperm motility at 5 and 10 min in a dose-dependent manner, with no significant difference at 15 and 30 min; sperm motility showed no significant difference on 5 -35 min exposure to naloxone at 0.004 -0.04 mg/L, nor on 5, 10, 15 and 20 min exposure at 0.4 -4 mg/L, but was significantly increased at 35 min. Compared with the corresponding concentrations of remifentanil alone, remifentanil + naloxone remarkably increased sperm motility at 0.1 + 0.004, 1 + 0.04, 10 + 0.4, and 100 + 4 mg/L at 5 and 10 min, with no obvious difference at 15 and 30 min. CONCLUSION: The onset and offset of the effect of remifentanil on sperm motility are rapid and its inhibitory effect can be antagonized by naloxone, which may be related with the micro-opioid receptor.

[Characterization of stromal cell cultures from the prostatic peripheral zone of men at different ages].

OBJECTIVE: To characterize age-related cellular phenotype alterations and growth rates of human prostatic stromal cell cultures from the normal prostatic peripheral zone of young donors (PZ-young) and old donors (PZ-old). METHODS: We isolated stromal cells from 10 donors of different ages, assessed the cellular phenotypes by immunocytostaining for prolyl-4-hydroxylase, alpha-smooth muscle actin (alpha-SMA) and desmin, and analysed the ultrastructure by transmission electron microscopy (TEM). The proliferation and apoptosis of the cells were determined by Cell Counting Kit-8 assay and flow cytometry, respectively. RESULTS: All the stromal cells were positive for prolyl-4-hydroxylase regardless of the donors' age, while alpha-SMA and desmin positive cells increased with their age. The positive expressions of alpha-SMA and desmin were (2.56 +/- 1.81)% and (0.89 +/- 0.93)% in PZ-young, and (38.89 +/- 11.22)% and (14.89 +/- 5.97)% in PZ-old (P < 0.01). The alpha-SMA- and/or desmin-positive stromal cells were morphologically large, flat and polygonal. Ultrastructural analysis showed that the cell cultures from PZ-old were richer in rough endoplasmic reticulum and golgi complexes. The stromal cells of PZ-old had a lower growth rate than that of PZ-young (P < 0.01), but there was no significant difference in the apoptosis rate between the two groups. CONCLUSION: Cellular phenotypes of human prostate stromal cell cultures change with the increase of age from predominantly typical fibroblasts to a mixture of fibroblasts and myofibroblasts, which might responsible for the high incidence of prostate cancer in elderly men.

Peripheral zone PSA density: a predominant variable to improve prostate cancer detection efficiency in men with PSA higher than 4 ng ml-1.

To improve the diagnostic efficiency of prostate cancer (PCa) and reduce unnecessary biopsies, we defined and analyzed the diagnostic efficiency of peripheral zone prostate-specific antigen (PSA) density (PZ-PSAD). Patients who underwent systematic 12-core prostate biopsies in Shanghai General Hospital (Shanghai, China) between January 2012 and January 2018 were retrospectively identified (n = 529). Another group of patients with benign prostatic hyperplasia (n = 100) were randomly preselected to obtain the PSA density of the non-PCa cohort (N-PSAD). Prostate volumes and transition zone volumes were measured using multiparameter magnetic resonance imaging (mpMRI) and were combined with PSA and N-PSAD to obtain the PZ-PSAD from a specific algorithm. Receiver operating characteristic (ROC) curve analysis was used to assess the PCa detection efficiency in patients stratified by PSA level, and the area under the ROC curve (AUC) of PZ-PSAD was higher than that of PSA, PSA density (PSAD), and transition zone PSA density (TZ-PSAD). PZ-PSAD could amend the diagnosis for more than half of the patients with inaccurate transrectal ultrasonography (TRUS) and mpMRI results. When TRUS and mpMRI findings were ambiguous to predict PCa (PIRADS score ≤3), PZ-PSAD could increase the positive rate of biopsy from 21.7% to 54.7%, and help 63.8% (150/235) of patients avoid unnecessary prostate biopsy. In patients whose PSA was 4.0-10.0 ng ml-1, 10.1-20.0 ng ml-1, and >20.0 ng ml-1, the ideal PZ-PSAD cut-off value for predicting clinically significant PCa was 0.019 ng ml-2, 0.297 ng ml-2, and 1.180 ng ml-2, respectively (sensitivity >90%). Compared with PSA, PSAD, and TZ-PSAD, the efficiency of PZ-PSAD for predicting PCa is the highest, leading to fewer missed diagnoses and unnecessary biopsies.

[Effects of inhaled anesthetics on human sperm motility in vitro].

OBJECTIVE: To study the effects of inhaled anesthetics on human sperm motility in vitro. METHODS: Sperm samples were obtained from 20 healthy men by masturbation and prepared by the swim-up technique. The effects of isoflurane and sevoflurane at the clinical concentration (1.4%-5.6%) and high concentration (5.6%-84%) on human sperm motility in vitro were observed at 25 degrees C by the computer-assisted sperm analysis (CASA). RESULTS: The sperm vitality and motility were significantly increased on 0.5-4 h exposure to isoflurane at the clinical concentration and decreased gradually at high concentration (42%-84%). The effect of isoflurane on human sperm motility and vitality at the clinical concentration was reversible when the anesthetic withdrawn. Sevoflurane had no effects on human sperm motility and vitality at either the clinical or high concentration. CONCLUSION: Isoflurane has a reversible increasing effect at the clinical concentration and a significant decreasing effect at the high concentration on the motility and vitality of human sperm, while sevoflurane does not affect human sperm motility and vitality at either concentration.

[Effects of isoflurane combined with diltiazem on human sperm motility].

OBJECTIVE: To observe the effects of isoflurane combined with diltiazem on human sperm motility in vitro and to investigate its possible mechanism. METHODS: Ten normal semen samples were collected, each divided into 9 groups, one as the control and the others treated in vitro with different concentrations of diltiazem or diltiazem +4.2% isoflurane for 1 hour. Sperm motility was observed with the computer-assisted sperm analyzer. RESULTS: Compared with the control, diltiazem significantly decreased sperm motility at the concentrations of 0.01 g/L, 0.04 g/L, 0.2 g/L and 1 g/L in a dose-dependent manner, and reduced it to approximately 0% at 1 g/L. When combined with 4.2% isoflurane, diltiazem obviously increased sperm motility at 0.01 g/L, markedly decreased it at 0.2 g/L, and effected no significant difference at 0.04 g/L and 1 g/L as compared with the corresponding concentrations of diltiazem alone. CONCLUSION: The stimulating effect of isoflurane on sperm motility may be associated with the calcium ion channel in sperm. When completely blocked by diltiazem, this effect may turn into an inhibition of sperm motility.

Deregulation of ATG9A by impaired AR signaling induces autophagy in prostate stromal fibroblasts and promotes BPH progression.

The activation of androgen receptor (AR) signaling plays an essential role in both prostate stromal cells and epithelial cells during the development of benign prostatic hyperplasia (BPH). Here we demonstrated that androgen ablation after 5α-reductase inhibitor (5-ARI) treatment induced autophagy in prostate stromal fibroblasts inhibiting cell apoptosis. In addition, we found that ATG9A expression was increased after androgen ablation, which facilitated autophagic flux development. Knockdown of ATG9A not only inhibited autophagy notably in prostate stromal fibroblasts, but also reduced the volumes of prostate stromal fibroblast and epithelial cell recombinant grafts in nude mice. In conclusion, our findings suggested that ATG9A upregulation after long-term 5-ARI treatment constitutes a possible mechanism of BPH progression. Thus, combined treatment with 5-ARI and autophagy inhibitory agents would reduce the risk of BPH progression.

MiR-185 attenuates androgen receptor function in prostate cancer indirectly by targeting bromodomain containing 8 isoform 2, an androgen receptor co-activator.

OBJECTIVES: Aberrant androgen receptor (AR) signaling functions are implicated in prostate cancer (PCa) pathogenesis. Here, we studied interactions between miR-185 and the bromodomain containing 8 isoform 2 (BRD8 ISO2) to investigate indirect mechanisms of miR-185 with respect to AR function through BRD8 ISO2 in PCa. METHODS: Putative miRNA response element (MRE) of miR-185 in 3'-untranslated region (3'-UTR) of BRD8 ISO2 mRNA was predicted by software and confirmed using dual-luciferase assays and Ago2 immunoprecipitation. BRD8 and AR expression were determined by qRT-PCR and Western blot in PCa cells and tissues. MMTV-Fluc reporter plasmids and dual-luciferase assays were used to evaluate AR activity. RESULTS: MRE prediction, dual-luciferase assays and Ago2 immunoprecipitation confirmed that miR-185 is capable of binding the 3'-UTR of BRD8 ISO2 mRNA. QRT-PCR and Western blot indicated that BRD8 ISO2 expression is decreased by miR-185 mimic transfection while increased by miR-185 inhibitor transfection. MMTV-Fluc reporter assays revealed that miR-185 can attenuate AR function by suppressing BRD8 ISO2. Additionally, Pearson's correlation analyses confirmed that BRD8 ISO2 mRNA expression is inversely correlated with miR-185 expression in clinical specimens. CONCLUSION: In addition to suppression of AR expression, miR-185 can attenuate AR function indirectly by suppressing BRD8 ISO2. MiR-185 and BRD8 ISO2 may be possible therapeutic targets for PCa treatment.

LIM domain only 2 over-expression in prostate stromal cells facilitates prostate cancer progression through paracrine of Interleukin-11.

Mechanisms of stromal-epithelial crosstalk are essential for Prostate cancer (PCa) tumorigenesis and progression. Peripheral zone of the prostate gland possesses a stronger inclination for PCa than transition zone. We previously found a variety of genes that differently expressed among different prostate stromal cells, including LIM domain only 2 (LMO2) which highly expressed in peripheral zone derived stromal cells (PZSCs) and PCa associated fibroblasts (CAFs) compared to transition zone derived stromal cells (TZSCs). Studies on its role in tumors have highlighted LMO2 as an oncogene. Herein, we aim to study the potential mechanisms of stromal LMO2 in promoting PCa progression. The in vitro cells co-culture and in vivo cells recombination revealed that LMO2 over-expressed prostate stromal cells could promote the proliferation and invasiveness of either prostate epithelial or cancer cells. Further protein array screening confirmed that stromal LMO2 stimulated the secretion of Interleukin-11 (IL-11), which could promote proliferation and invasiveness of PCa cells via IL-11 receptor α (IL11Rα) - STAT3 signaling. Moreover, stromal LMO2 over-expression could suppress miR-204-5p which was proven to be a negative regulator of IL-11 expression. Taken together, results of our study demonstrate that prostate stromal LMO2 is capable of stimulating IL-11 secretion and by which activates IL11Rα - STAT3 signaling in PCa cells and then facilitates PCa progression. These results may make stromal LMO2 responsible for zonal characteristic of PCa and as a target for PCa microenvironment-targeted therapy.

Discrepancy of blood pressure between the brachial artery and radial artery.

BACKGROUND: In this study, we attempted to find the relations between blood pressure (BP) measured on the brachial artery (bBP) and BP assessed on the radial artery (rBP) in the right arm. METHODS: Three hundred and fifteen patients were enrolled in this study. Those who had peripheral vascular disease, wounds of arm skin or subcutaneous tissue infection were excluded. After a 15-minute equilibration and stabilization period after inducation of anesthesia, three bBP and rBP records were obtained sequentially using an oscillometric device with an adult cuff and infant cuff, respectively. Order for each BP was randomized. RESULTS: The bBP was significantly lower than the rBP (P<0.05). The difference between the two values varied from 13 to 18 mmHg in systolic BP (SBP), diastolic BP (DBP) and mean blood pressure (MAP) respectively. And the rBP was positively correlated with the bBP (r=0.872, 0.754, 0.765; P<0.001, <0.001, <0.001; SBP, DBP, MAP, respectively). CONCLUSION: The bBP value can be evaluated by the noninvasive measurements of rBP using an appropriate cuff in clinical practice.

Androgen receptors expressed by prostatic stromal cells obtained from younger versus older males exhibit opposite roles in prostate cancer progression.

Aging is a major risk factor for prostate cancer (PCa), and prostatic stromal cells may also promote PCa progression. Accordingly, stromal cells do not equally promote PCa in older males and younger males. Therefore, it is also possible that the expression of androgen receptors (ARs) by prostatic stromal cells in older versus younger males plays different roles in PCa progression. Using a gene knockdown technique and coculture system, we found that the knockdown of the AR in prostatic stromal cells obtained from younger males could promote the invasiveness and metastasis of cocultured PC3/LNCaP cells in vitro. By contrast, the invasiveness and metastasis of LNCaP cells was inhibited when cocultured with prostatic stromal cells from older males that when AR expression was knocked down. Moreover, after targeting AR expression with small hairpin RNA (shRNA), matrix metalloproteinase (MMP) expression in stromal cells was observed to increase in the younger group, but decreased or remained unchanged in the older group. One exception, however, was observed with MMP9. In vivo, after knocking down AR expression in prostatic stromal cells, the incidence of metastatic lymph nodes was observed to increase in the younger age group, but decreased in the older age group. Together, these data suggest that the AR in prostatic stromal cells played opposite roles in PCa metastasis for older versus younger males. Therefore, collectively, the function of the AR in prostatic stromal cells appears to change with age, and this may account for the increased incidence of PCa in older males.

Primary stromal cells isolated from human various histological/pathological prostate have different phenotypes and tumor promotion role.

BACKGROUND: Prostate stromal cells are known to regulate epithelial growth as well as support and maintain epithelial function. However, how stromal cells regulate epithelial cells and what differences among various histological/pathological prostate stromal cells in prostate cancer progression still remain unclear. This study aimed to investigate the different phenotypes of human various histological/pathological prostate stromal cells, and their role in tumor promotion. METHODS: The different phenotypes of the human normal prostatic peripheral zonal primary stromal cells (NPPF), transitional zonal primary stromal cells (NPTF), and prostate cancer associated primary stromal cells (CAF) were examined with growth curves and Annexin V-fluorescein isothiocyanate (FITC) assay. The different effects on prostate cancer cell line C4-2B by NPPF, NPTF, and CAF were examined with MTT assay and Annexin V-FITC assay. The gene expression of different histological/pathological prostate stromal cells was profiled by microarray and hierarchical cluster analysis. RESULTS: The growth rate of NPPF, NPTF and CAF gradually increased, followed by decreasing apoptosis. In vitro stromal-C4-2B cell line co-culture models, the proliferation and apoptosis of C4-2B cell line were differently affected by human various histological/pathological prostate stromal cells. CAF showed the most powerful effect to C4-2B cell line, as opposed to a weakest effect of NPTF. Microarray and hierarchical cluster analysis showed that the differentially expressed genes of CAF and NPPF were less than NPPF and NPTF, or CAF and NPTF. This was consistent with clinical observations that prostate cancer mostly derived from the peripheral zone and does not usually occur in the transitional zone. CONCLUSION: NPPF, NPTF and CAF possess extremely different biological characteristics and gene expression, which may play an important role in genesis and development of prostate cancer.

Differences in phenotype and gene expression of prostate stromal cells from patients of varying ages and their influence on tumour formation by prostate epithelial cells.

Prostate cancer (PCa) is an age-related disease, and the stromal microenvironment plays an important role in prostatic malignant progression. However, the differences in prostate stromal cells present in young and old tissue are still obscure. We established primary cultured stromal cells from normal prostatic peripheral zone (PZ) of donors of varying ages and found that cultured stromal cells from old donors (PZ-old) were more enlarged and polygonal than those from young donors (PZ-young). Furthermore, based on immunocytochemical and ultrastructural analysis, the components of stromal cells changed from a majority of fibroblasts to a mixture of fibroblasts and myofibroblasts with increasing donor age. Using a three-dimensional in vitro culture system, we found that PZ-old stromal cells could enhance the proliferation, migration and invasion of cocultured benign BPH-1 and PC-3 cells. Using an in vivo tissue recombination system, we also found that PZ-old stromal cells are more effective than PZ-young cells in promoting tumour formation by BPH-1 cells of high passage (>100) and PC-3 cells. To probe the possible mechanism of these effects, we performed cDNA microarray analysis and profiled 509 upregulated genes and 188 downregulated genes in PZ-old cells. Among the changed genes, we found genes coding for a subset of paracrine factors that are capable of influencing adjacent epithelial cells; these include hepatocyte growth factor (HGF), fibroblast growth factor 5 (FGF5), insulin-like growth factor 2 (IGF2), insulin-like growth factor-binding protein 4 (IGFBP4), IGFBP5 and matrix metallopeptidase 1 (MMP1). Changes in the expression of these genes were further confirmed by quantitative real-time polymerase chain reaction (PCR), Western blotting and enzyme-linked immunosorbent assays. Overall, our findings indicate that stromal cells from prostate PZ of old donors are more active than similar cells from young donors in promoting the malignant process of adjacent epithelial cells. This finding hints at a new potential strategy for the prevention of PCa.

Slow release properties and liver-targeting characteristics of methotrexate erythrocyte carriers.

To study the releasing properties and tissue-targeting characteristics of methotrexate-loaded red blood cells (MTX-RBCs), pharmacokinetics and tissue distributions of intravenous injected MTX-RBCs and free MTX were compared. MTX-RBCs were made from rat erythrocytes using a hypertonic method. After i.v. injection of MTX-RBCs or free MTX to rats, both plasma and tissue homogenate samples at each time-point were collected and analyzed by RP-HPLC. From this data, pharmacokinetic parameters and tissue distributions of MTX were obtained. MTX-RBCs were successfully produced by the hypertonic method. After i.v. injection, MTX-RBCs displayed more than three times longer half-life and MRT than free MTX, and the velocity of MTX clearance from plasma was much slower. The ratio of area under the concentration time curve (AUC)(tissue) to AUC(plasma) in the liver was clearly higher than that for other organs after MTX-RBCs administration; MRT in the liver was also longer. This study has demonstrated that the hypertonic method for making MTX-RBCs has led to a preparation with slow release properties as well as liver-targeting characteristics in rats. This approach offers considerable potential for the treatment of tumors in liver, which would merit further investigation.