OBJECTIVETo study the inhibitory effect of matrine on the proliferation of the prostate cancer cell line LNCaP and the expression of the androgen receptor (AR).

METHODSLNCaP cells were treated with matrine at the concentration of 0.5, 1.0, 1.5, 2.0 and 3.0 g/L for 12, 24 and 36 hours, the cell growth activity determined by MTT colorimetry and trypan blue staining at 36 hours, the cell cycle changes detected by flow cytometry and the expression of AR by Western blot at 24 hours.

RESULTSMatrine suppressed the in vitro growth of the androgen-sensitive prostate cancer cell line LNCaP in a time- and dose-dependent manner, blocked the cell cycles in the G2/M phase and decreased the expression of AR in the cell line in a dose-dependent manner (P < 0.01).

CONCLUSIONMatrine can significantly inhibit the in vitro growth of NCaP cells by down-regulating the expression of AR and blocking cell cycles.

Alkaloids ; pharmacology ; Blotting, Western ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Flow Cytometry ; Humans ; Male ; Prostatic Neoplasms ; metabolism ; pathology ; Quinolizines ; pharmacology ; Receptors, Androgen ; metabolism

OBJECTIVEThis study was to explore the cytotoxic effect and the related injury mechanism of deoxynivalenol (DON) on articular chondrocytes in human embryo.

METHODSArticular cartilage cells were isolated from knees of human embryo and cultured in DMEM/F12 medium. The cells of the 4th generation were divided into five groups and incubated with varying concentrations of DON as the followings: control group and group with DON of 0.1, 0.2, 0.4, 1.0 µg/ml. The effects of DON were observed 72 hours after incubation. Cell apoptosis was assayed by flow cytometry (FCM) with Annexin V-FITC/PI staining; MMP-13 and PGE2 were detected by ELISA kits; NO was measured by Griess assay with spectrophotometer. Inducible nitric oxide synthase (iNOS) and collagen II in cells were detected by FCM. The expression levels of iNOS, mRNA and collagen II mRNA were measured with RT-PCR.

RESULTSThe rates of cell apoptosis in DON groups were 6.78% - 19.05%, which were significantly higher than that in control (1.20%, F = 174.761, P < 0.05). The levels of NO in DON groups were 20.8 - 40.7 µmol/L, which were significantly higher than that in control (10.2 µmol/L, F = 91.966, P < 0.05). The levels of MMP-13 in DON groups were 0.25 - 0.56 µmol/L, which were significantly higher than that in control (0 µmol/L, F = 78.420, P < 0.05). The levels of PGE2 in DON groups were 3.2-20.6 µmol/L, which were significantly higher than that in control (11.6 µmol/L, F = 276.453, P < 0.05). The proportions of cells with positive iNOS in DON groups were 14.8% - 56.8% which were significantly higher than that in controls (7.1%, F = 214.614, P < 0.05). The proportions of cells with positive collagen II in groups with DON of 0.4 µg/ml and 1.0 µg/ml were 56.7% and 52.7%, which were significantly lower than that in control (62.2%, F = 5.134, P < 0.05). The relative absorbance values of iNOS mRNA in DON groups were 1.07 - 1.33, which were significantly higher than that in control (0.62, F = 8.358, P < 0.05). The levels of collagen II mRNA in groups with DON of 0.4 µg/ml and 1.0 µg/ml were 0.83 and 0.82, which were significantly lower than that in control (1.14, F = 7.887, P < 0.05).

CONCLUSIONDON could promote anabolism of NO in articular cartilage cells by which up-regulated the expression of PGE2 and MMP-13, which both promoted resolution of articular cartilage matrix such as collagen II. DON induced apoptosis in articular cartilage cells.

Cartilage, Articular ; cytology ; embryology ; Cells, Cultured ; Chondrocytes ; drug effects ; metabolism ; Dinoprostone ; metabolism ; Humans ; Matrix Metalloproteinase 13 ; metabolism ; Nitric Oxide ; biosynthesis ; Trichothecenes ; toxicity

OBJECTIVETo investigate the seasonal characteristics and the sources of elements and ions with different sizes in the aerosols in Beijing.

METHODSSamples of particulate matters (PM2.5), PM10, and total suspended particle (TSP) aerosols were collected simultaneously in Beijing from July 2001 to April 2003. The aerosol was chemically characterized by measuring 23 elements and 18 water-soluble ions by inductively coupled plasma-atomic emission spectroscopy (ICP-AES) and ion chromatography (IC), respectively.

RESULTSThe samples were divided into four categories: spring non-dust, spring dust, summer dust, and winter dust. TSP, PM10, and PM2.5 were most abundant in the spring dust, and the least in summer dust. The average mass ratios of PM > 10, PM2.5-10, and PM2.5 to TSP confirmed that in the spring dust both the large coarse (PM > 10) and fine particles (PM2.5) contributed significantly in summer PM2.5, PM2.5-10, and PM > 10 contributed similar fractions to TSP, and in winter much PM2.5. The seasonal variation characteristics of the elements and ions were used to divide them into four groups: crustal, pollutant, mixed, and secondary. The highest levels of crustal elements, such as Al, Fe, and Ca, were found in the dust season, the highest levels of pollutant elements and ions, such as As, F-, and Cl-, were observed in winter, and the highest levels of secondary ions (SO4(2-), NO3-, and NH4+) were seen both in summer and in winter. The mixed group (Eu, Ni, and Cu) showed the characteristics of both crustal and pollutant elements. The mineral aerosol from outside Beijing contributed more than that from the local part in all the reasons but summer, estimated using a newly developed element tracer technique.

Aerosols ; China ; Chromatography, Ion Exchange ; Environmental Monitoring ; Particle Size ; Particulate Matter ; analysis ; chemistry ; Seasons ; Spectrophotometry, Atomic

OBJECTIVETo assess the diagnostic value and potentially protective capacity of tumor necrosis factor-alpha (TNF-alpha), interleukin 1beta (IL-1beta) and heat-shock protein 70 (HSP70) in chronic bacterial prostatitis (CBP) and chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS).

METHODSWe determined the levels of cytokines TNF-alpha, IL-1beta and HSP70 by ELISA in the seminal plasma of 150 men: 36 with CBP, 43 with CP/CPPS IIIA, 46 with CP/CPPS IIIB, and 25 healthy controls. We analyzed the correlation of the HSP70 expression in the CBP and CP/CPPS patients with the chronic prostatitis symptom index (CPSI).

RESULTSSignificantly increased levels of TNF-alpha, IL-1beta and HSP70 were observed in the seminal plasma of the CBP patients as compared with the CP/CPPS patients and healthy controls. The expression of IL-1beta was significantly higher in the patients with CP/CPPS IIIA than in those with CP/CPPS III B and the controls, while the HSP70 level remarkably lower in those with CP/CPPS than in the controls, and its concentration in the seminal plasma of the CBP patients was negatively correlated with CPSI.

CONCLUSIONThe levels of HSP70 and IL-1beta in the seminal plasma appear to be most reliable molecular biological markers for the diagnosis of CBP and CP/CPPS, respectively. HSP7O has an important protective role in the regulation of cell functions in CBP patients. CP/CPPS is probably detrimental to the function of T cells and consequently suppresses the expression of HSP70.

Adolescent ; Adult ; Biomarkers ; metabolism ; Case-Control Studies ; Cytokines ; metabolism ; HSP70 Heat-Shock Proteins ; metabolism ; Humans ; Male ; Middle Aged ; Pelvic Pain ; metabolism ; Prostatitis ; metabolism ; Semen ; metabolism ; Young Adult

OBJECTIVETo investigate the effect of the mammalian target of rapamycin (mTOR) inhibitor CCI-779 on the chemosensitivity of androgen-independent prostate cancer cell line PC-3.

METHODSProstate cancer cells PC-3 were cultured and treated with CCI-779, Paclitaxel and combination of the two. Then the inhibitory effects of the three medications on the growth of the PC-3 cells were determined by MTT, and the their cell cycle and apoptosis were detected by flow cytometry.

RESULTSCompared with the control group, the three medications all significantly inhibited the proliferation of the PC-3 cells, and the combined method even enhanced the effect. Flow cytometry showed that CCI-779 and Paclitaxel blocked the cell cycle mainly in the G1/G2 stage, while the combined medication mainly in the G0/G1 stage. Significantly increased apoptosis of the PC-3 cells was observed in the three medication groups as compared with the control group (P < 0.01).

CONCLUSIONCCI-779 can inhibit the proliferation of PC-3 cells and enhance the chemosensitivity of prostate cancer.

Antineoplastic Agents ; pharmacology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Drug Therapy, Combination ; Humans ; Male ; Paclitaxel ; pharmacology ; Prostatic Neoplasms ; drug therapy ; Protein Kinase Inhibitors ; pharmacology ; Sirolimus ; analogs & derivatives ; antagonists & inhibitors ; pharmacology

BACKGROUND: Studies have shown that bone marrow mesenchymal stem cells have the potential of differentiation into alveolar epithelial cells in vitro, but so far no study has indicated that adipose-derived mesenchymal stem cells (ADSCs) can be differentiated into alveolar epithelial cells through long-term Transwell co-culture. OBJECTIVE: To observe whether rat lung epithelial-T-antigen negative cell lines (RLE-6TN) can induce rat ADSCs to differentiate into type II alveolar epithelial cells by long-term Transwell co-culture. METHODS: Three SPF health female Sprague-Dawley rats were used as donors to separate, extract, culture and identity ADSCs. The experimental group was subjected to the Transwell co-culture of ADSCs and RLE-6TN, while the control group was subjected to the culture of ADSCs alone. The morphological changes of ADSCs were observed by the inverted phase contrast microscope at 21 days after co-culture. Immunofluorescence staining using surfactant protein C (SP-C) was performed on the co-cultured ADSCs. The fluorescence staining was observed using the inverted fluorescence microscope. Integral optical density (IOD) analysis was conducted by Image pro plus 6.0 software. RESULTS AND CONCLUSION: RLE-6TN cells were identified by fluorescence staining with stable expression of SP-C protein (red fluorescence) in the experimental group, and there was no red fluorescence in the control group. After 21-day co-culture, the cell shape in the experimental group was transformed from the long spindle shape into oval or polygon shape gradually, while the cell shape in the control group remained fibroblast-like. These results show that RLE-6TN can induce ADSCs to differentiate into type II alveolar epithelial cells after a long-term (21 days) co-culture.

Objective To assess the efficacy of two antibiotic prophylactic regimens in a prospective randomized trial in 1 year for patients undergoing insertion of catheters,and to provide the evidence for uniform consensus existing on the timing,route,and choice of antibiotic.Methods During a period of 12 months,78 patients,who consecutively entered the peritoneal dialysis programme,[45 women and 33 men,mean age (48.2?15.7)years] were included.The prophylactic regimens were a single dose of ceftriaxone (1.0 g) given intravenously 30 minutes before surgery (Group A) and given cefazolin (0.25 g/L) i.p.in the each dialysis bag for 3 days postoperatively (Group B).All operations were performed in one room.The wound was observed every day,and body temperature,Count of white blood corpuscle and type,dialysate were examined every day. Results In Group A and B,none of the patients showed peritonitis or wound infection during the post-operative period (within 10 days).One of 39 patients(2.5%) in the group A,and 2 of 39 patients (5.1%) in the group B had exit site infection (P＞0.05).Conclusions There is no significant difference in the incidence of peritonitis and wound infection between two groups. Prophylactic preoperative single-dose antibiotics intravenously do as well as antibiotics given intraperitoneally for peritoneal dialysis catheter insertion,but is much more convenient.

AIMTo study the influences of dibutyryl cyclic adenosine monophosphate (dbcAMP) and forskolin on human sperm motility in vitro.

METHODSSemen samples, aseptically obtained by masturbation and prepared by swim-up technique from 20 fertile men, were incubated with different concentrations of dbcAMP and forskolin at 37 deg. Measurements were carried out after 10 min, 20 min, 30 min and 60 min incubation. Motility parameters were estimated by using an automatic analyzing system.

RESULTSTreatment with dbcAMP or forskolin resulted in a significant increase in sperm motility and progressive motility. The larger the concentrations of dbcAMP or forskolin, the greater the effect appeared. The straight linear velocity and curvilinear velocity were not affected by both agents.

CONCLUSIONdbcAMP and forskolin increase the motility and progressive motility of human sperm in vitro.

Adult ; Bucladesine ; administration & dosage ; pharmacology ; Colforsin ; administration & dosage ; pharmacology ; Dose-Response Relationship, Drug ; Humans ; In Vitro Techniques ; Male ; Osmolar Concentration ; Sperm Motility ; drug effects

OBJECTIVETo examine the levels of protein oxidation in systemic lupus erythematosus (SLE) and to evaluate the relation between oxidative protein damage and disease activity index.

METHODSPlasma was collected from SLE patients and healthy subjects as controls. Protein-bound carbonyls, protein thiols, superoxide dismutase (SOD), myeloperoxidase (MPO) and total antioxidant capacity (TAOC) were determined by spectrophotometry. Levels of anti-double-stranded DNA (anti-dsDNA) and antinuclear antibody (ANA) were quantified by enzyme-linked immunosorbent assay.

RESULTSWhen comparing with the control subjects, SLE patients exhibited elevated levels of protein carbonyls [(0.101 +/- 0.033) nmol/mg, (0.061 +/- 0.019)nmol/mg, P < 0.01], degraded levels of protein thiols [(3.911 +/- 0.968) nmol/mg, (4.655 +/- 0.798) nmol/mg, P < 0.01] and activities of T-SOD [(67.01 +/- 12.22) U/ml, (97.35 +/- 14.11) U/ml, P < 0.01]. Levels of plasma protein thiols and activities of T-SOD were lower in SLE patients positive for anti-dsDNA antibody, compared with patients negative for anti-dsDNA antibody.

CONCLUSIONWe found that the elevated levels of multiple markers of protein oxidation and degraded activities of antioxidant enzymes in plasma from SLE patients existed when comparing with the controls, and the all the levels were correlated with disease activity. Our findings suggested that protein oxidation might play a role in the pathogenesis of chronic organ damage in SLE.

Adult ; Antibodies, Antinuclear ; metabolism ; DNA ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Lupus Erythematosus, Systemic ; blood ; metabolism ; Male ; Middle Aged ; Oxidation-Reduction ; Peroxidase ; metabolism ; Proteins ; metabolism ; Superoxide Dismutase ; metabolism ; Young Adult

OBJECTIVETo investigate the curcumin-induced the expression of IkappaBalpha in androgen-dependent (LNCaP) and androgen-independent (PC3) prostate cancer cells, and to study the mechanisms of curcumin on the proliferative inhibition of prostate cancer cells.

METHODSAfter LNCaP and PC3 cells were affected by 10, 25, 50, 75, 100 micromol/L curcumin respectively, the cell activity was assayed with methyl thiazolyl tetrazolium (MTT) method at 5, 12 and 24 hours; Flow cytometry was adopted to observe the cell cycle of LNCaP and PC3 cells at 24 hours. After 5 hours, the expression of IkappaBalpha in LNCaP and PC3 cells was observed with Western blotting.

RESULTSCurcumin obviously suppressed the proliferation of LNCaP and PC3 cells in does-dependent and time-dependent manners. Curcumin could arrest the cell cycle of LNCaP and PC3 cells at G(2), M phase and then induce cell apoptosis. The expression of IkappaBalpha in LNCaP cells had no significant difference after using curcumin (F = 0.129, P > 0.05). However, the expression of IkappaBalpha in PC3 cells increased gradually with the inducement of concentration-increased curcumin (F = 31.618, P < 0.05).

CONCLUSIONSIkappaBalpha may play a role in the curcumin inducing apoptosis of PC3 cell, while the curcumin inducing apoptosis of LNCaP cells is by antioxidation and inhibiting metabolites formation in LNCaP cells.

Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Curcumin ; pharmacology ; Dose-Response Relationship, Drug ; Humans ; I-kappa B Proteins ; biosynthesis ; Male ; NF-KappaB Inhibitor alpha ; Prostatic Neoplasms ; metabolism ; pathology