Purpose:To determine the efficacy of paclitaxel in human bladder cancer lines and to investigate the mechanism by which paclita xel induce apoptosis in human bladder cancer cells. Methods:BIU-87, 5637, T24 and EJ bladder cancer cell lines wer e cultured by techniques of cell culture in vitro. The cytotoxic activity an d apoptosis induction abilities of paclitaxel were analyzed by MTT and Annexin- V assay as well as DNA cytometry , respectively. The effects on the cell cycle w ere assessed by flow cytometry of propidium iodide. The expressions of Bcl-2, B ax, p53 and Caspase3 proteins were determined by flow cytometry immunofluorescen ce. Results:Paclitaxel dose-dependent inhibition of cell prolifera tion was seen.Paclitaxel induced G_2/M arrest (71.29% and 64.57%) which was maximal in 5637 and EJ cell lines. While paclitaxel at 1?g/ml concentration ex posure to 5637 12h, 14h and 48h respectively, the apoptosis rates of the respect ive times were 5.0%, 12.9%, 27.6%. The expression of genes p53 and Bcl-2 was no t influenced, whereas the expression of Bax and Caspase3 had increases time-dep endently after exposure to paclitaxel. The analysis of Annexin-V showed a drama tic dose-dependent increase of apoptosis. Conclusions:Paclitaxel inhibited bladder cancer cells prolifera tion and had more effect on those cells whose grade was lower and doubling time was longer. Paclitaxel could block G_2/M arrest, and induce apoptosis by th e path of Bcl-2/Bax in bladder cancer cell lines.

OBJECTIVESTo investigate the androgen receptor (AR) isoforms expression in human benign and malignant prostatic tissues and LNCaP cells.

METHODSUsing high resolution isoelectric focusing (IEF), the different expression of AR isoforms were demosntrated in human benign and malignant prostatic tissues and LNCaP cells.

RESULTSData were obtained from 41 AR-positive BPH, three prostatic cancer specimens, and LNCaP cells. From these materials, three types of AR isoforms were detected with pI values at 6.5, 6.0 and 5.3. In the case of BPH tissues, 15 (36.5%) specimens expressed all the three types of isoforms at pI 6.5, 6.0 and 5.3, and 10 (24.4%) samples contained isoforms at pI 6.5 and 5.3, five (12.2%) samples indicated isoforms at pI 6.5 and 6.0, four (9.8%) showed the isoforms at pI 6.0 and 5.3. Of all the 41 specimens, two (4.9%) and two (4.9%) as well as three (7.3%) denoted the isoforme at pI 6.5, 6.0 and 5.3 respectively. As for three prostatic cancer specimens, one sample showed all the three types of AR isoforms at pI 6.5, 6.0, 5.3, but another specimen expressed at pI 6.5 and 6.0, and only one failed to indicate any types of isoforms. LNCaP cells expressed all three types of AR isoforms at pI 6.5, 6.0 and 5.3. Binding of 3H-dihydrotestosterone to these three types of isoforms was inhibited by the addition of 100-fold excess of DHT and testosterone. No effect of progesterone, oestradiol and diethylstilboestrol on tritiated hormone binding was observed.

CONCLUSIONSThe expression of AR isoforms is different among various patients and different between BPH and LNCaP cells, though no clear explanation could be induced for this. These results suggest the possibility of explaining effective hormonal therapy to prostatic disease in the future.

Humans ; Male ; Prostate ; metabolism ; Protein Isoforms ; metabolism ; Receptors, Androgen ; metabolism ; Tumor Cells, Cultured

Apoptosis is an essential factor in keeping homeostasis of the organism. Apoptosis is regulated by a series of cytokines. Bcl-2 family proteins are key regulators of apoptosis. The Bcl-2 family includes both anti- and pro-apoptotic proteins with opposing biological functions. Their interaction regulates the transmission of the apoptosis signal. High expression of anti-apoptotic members such as Bcl-2 and Bcl-xL are commonly found in human cancers. In recent years, following the disclosing of the crystal structures of Bcl-2 family proteins, researchers have paid attention to the development of the small molecule inhibitors of Bcl-2 family proteins. This article reviews the progress in this field from the view of drug design.
Antimycin A ; chemistry ; pharmacology ; Antineoplastic Agents ; chemistry ; pharmacology ; Apoptosis ; drug effects ; Benzopyrans ; chemistry ; pharmacology ; Biphenyl Compounds ; chemistry ; pharmacology ; Cell Line, Tumor ; Drug Design ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Gossypol ; chemistry ; pharmacology ; Humans ; Nitriles ; chemistry ; pharmacology ; Nitrophenols ; chemistry ; pharmacology ; Piperazines ; chemistry ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; antagonists & inhibitors ; pharmacology ; Structure-Activity Relationship ; Sulfonamides ; chemistry ; pharmacology ; Thiazoles ; chemistry ; pharmacology ; bcl-X Protein ; antagonists & inhibitors ; pharmacology

Chronic prostatitis (CP)/chronic pelvic pain syndrome (CPPS) is a common problem of medically controversial condition that causes considerable morbidity and impact on life. Although there are many competing causes proposed, the etiology and pathogenesis of CP/CPPS remain unclear. The causative factors underlying the CPPS are not fully understood. The optimal management of CP/CPPS is still unknown. The guideline of diagnosis and management of CP/CPPS based on evidence base medicine is not yet established. Many problems are still not resolved, such as the significance of leukocytes and the role of inflammation in CP/CPPS, the significance of bacteria presence and the role of infection in CP/CPPS, the correlation between leukocytes/bacteria and severity of symptoms, how to divide the subgroups of CP/CPPS, the role of antimicrobial therapy in the treatment of men with CP/CPPS, why patients with category IIIb complain of symptoms, while those with category IV complain of none. Although CP/CPPS is now achieving greater recognition, well-designed studies with large sample size should be performed.
Chronic Disease ; Humans ; Male ; Pelvic Pain ; etiology ; microbiology ; therapy ; Prostatitis ; etiology ; microbiology ; therapy

OBJECTIVETo investigate the protective effect of melatonin on residual hair follicle cells of scald rats at early stage.

METHODSEighteen male Sprague-Dawley rats were randomly divided into scald group, treatment group, sham group , with 6 rats in each group. The rats in scald group and treatment group were subjected to 30% TBSA partial thickness scald on the back, and were resuscitated with balanced solution after 1 hour, while those in sham group were immersed in water at 37 degrees C for 25 s to simulate scald, and did not receive fluid replacement. Rats in treatment group were intraperitoneally injected with 10 mg/kg melatonin solution at 1 minute, 8 hours and 12 hours after scald, while those in sham group and scald group were given equal volume of 1% alcohol sodium-isotonic saline instead. Tissue samples were harvested at 6, 12 and 24 post scald hours (PSH) for determination of MDA and GSH levels. Apoptosis of residul hair follicle was detected by TUNEL method and immunohistochemistry of caspase-3.

RESULTSThe level of MDA in scald group at each time point was much higher than that in sham group (P < 0.01) and treatment group (P < 0.05), and it peaked at 12 PSH. The changes in GSH were just opposite to that of MDA. Under fluorescence microscope, the residual hair follicle cells were blue, and the apoptotic cells appeared green. The apoptosis rate in scald group at 6, 12, 24 PSH was obviously higher than that in sham (P < 0.01) and treatment groups (P < 0.05), which was (20.2 +/- 3.4)% vs (4.3 +/- 2.3)% vs (10.9 +/- 3.2)%, (31.2 +/- 3.6)% vs (5.1 +/- 2.5)% vs (19.1 +/- 3.7)%, (22.4 +/- 2.7)% vs (4.1 +/- 2.4)% vs (13.1 +/- 3.4)%, respectively. The score of caspase-3 positive cell in scald group was higher than those in sham group (P < 0.01) and treatment group (P < 0.05).

CONCLUSIONSThere is obvious correlation between oxidative stress and apoptosis rate of hair follicle cells in rats with partial thickness scald. Early administration of melatonin may have anti-apoptosis ability for residual hair follicle cells by attenuation of oxidative stress.

Animals ; Apoptosis ; Burns ; drug therapy ; metabolism ; Hair Follicle ; cytology ; metabolism ; Male ; Melatonin ; therapeutic use ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley

Hypoxia-induced 15-hydroxyeicosatetraenoic acid (15-HETE) is an essential mediator to constrict pulmonary arteries (PA). The signaling pathway involved in 15-HETE-induced PA vasoconstriction remains obscure. The aim of the present study was to test the hypothesis that hypoxic PA constriction induced by 15-HETE was possibly regulated by the extracellular signal-regulated kinase-1/2 (ERK1/2) pathway. PA ring tension measurement, Western blot and immunocytochemistry were used in the study to determine the possible role of ERK1/2 in 15-HETE-induced PA vasoconstriction. The organ bath for PA rings tension study was employed. Adult male Wistar rats were raised in hypoxic environment with fractional inspired oxygen (FIO2, 0.12) for 9 d. PA 1~1.5 mm in diameter were dissected and cut into 3 mm long rings for tension study. ERK1/2 up-stream kinase (MEK) inhibitor PD98059, which blocks the activation of ERK1/2, was used. The results showed that pretreatment of PD98059 significantly blunted 15-HETE-induced PA vasoconstrictions in the rings from hypoxic rat. Moreover, in endothelium-denuded rings, PD98059 also significantly attenuated 15-HETE-induced vasoconstriction. Phosphorylation of ERK1/2 in pulmonary arterial smooth muscle cells (PASMCs) of rat was enhanced evidently when stimulated by 15-HETE. Thus, the data suggest that ERK1/2 signaling pathway is involved in 15-HETE-induced hypoxic pulmonary vasoconstriction.
Animals ; Flavonoids ; pharmacology ; Hydroxyeicosatetraenoic Acids ; antagonists & inhibitors ; pharmacology ; Hypoxia ; physiopathology ; MAP Kinase Signaling System ; physiology ; Male ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; drug effects ; Pulmonary Artery ; cytology ; drug effects ; physiopathology ; Rats ; Rats, Wistar ; Vasoconstriction ; drug effects

AIMTo study upon to serum deprivation if delta-opioid receptor activation has direct effect on cultured impaired cardiomyocytes survival.

METHODSMyocardial cells of neonatal rats were cultured in vitro. The cell viability was determined with crystal violet staining uptake. The percentage of S + G2 + M in cell cycle was determined by flow cytometry. Apoptosis rates were determined by flow cytometry (FCM). The expression of Caspase-3 were investigated by Western blotting.

RESULTSMyocardial cells of neonatal rats were cultured of serum-free in vitro, apoptotic index was significantly increased, the expression of Caspase-3 was significantly increased, free-serum induced apoptosis in cardiac myocytes after 48 h. At concentrations of 10 nmol x L(-1) - 10 micromol x L(-1), a delta opoid receptor agonist [D-Ala2, D-Leu5]-enkephalin DADLE promoted the myocardial cells survival, in a concentration-dependent manner. The optimal response was achieved at 0.1 micromol x L(-1), which increase survival index of cardiac myocyte, percentage of S + G2 + M in cell cycle, decrease apoptotic index of cardiac myocyte, and the expression activate caspase-3. Delta-opioid receptor antagonist naltrindole at 10 micromol x L(-1) inhibited the promoting effects of DADLE, which decrease survival index of cardiac myocyte, and percentage of S + G2 + M in cell cycle, increase apoptotic index of cardiac myocyte and the expression of Caspase-3.

CONCLUSIONThe protective of delta-opioid receptor activation can promote survival in cultured impaired myocardial cells.

Animals ; Animals, Newborn ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Survival ; drug effects ; Cells, Cultured ; Culture Media, Serum-Free ; Enkephalin, Leucine-2-Alanine ; pharmacology ; Female ; Male ; Myocytes, Cardiac ; cytology ; Rats ; Rats, Sprague-Dawley ; Receptors, Opioid, delta ; agonists

BACKGROUNDThis study was to evaluate whether anergic cells induced by the blockade of CD40-CD154 and CD28-B7 costimulatory pathways can act as potent immunoregulatory cells in vitro and prolong cardiac allograft survival after adoptive transfer.

METHODSAnergic cells were induced in vitro by the addition of anti-CD154 and anti-CD80 monoclonal antibodies (mAbs) to primary MLR (mixed lymphocyte reaction) consisting of BALB/c as responder and C3H as stimulator. Anergic cells were added to a newly formed MLR in assessing the regulatory capacity and antigen specificity of anergic cells. The ability of anergic cells to respond to antigen and/or exogenous recombinant mouse interleukin-2 (rmIL-2) was tested. For in vivo studies, anergic cells were intravenously injected into 3.0-Gy gamma-irradiated BALB/c mice immediately after heterotopic abdominal cardiac transplantation. To prolong allograft survival, recipient mice injected with anergic cells received rapamycin therapy [1 mg.day(-1).kg(-1)].

RESULTSAnergic cells strongly suppressed the proliferation of naicaron;ve BALB/c splenocytes against the original (C3H) stimulator in a dose-dependent manner, but they failed to suppress the proliferation of naicaron;ve BALB/c splenocytes against the third-party (C57BL/6J) stimulator. The anergic state was reversed by both original (C3H) stimulator and additional exogenous IL-2. In in vivo studies, untreated irradiated BALB/c mice rejected C3H cardiac allografts with a mean survival time of (8.6 +/- 1.1) days, whereas those injected with the anergic cells rejected the allografts with a mean survival time of (11.8 +/- 1.9) days, which was slightly longer than that of the untreated mice. The protocol based on anergic cells injection plus rapamycin therapy could prolong allograft survival significantly [(29.6 +/- 4.4) days].

CONCLUSIONSAnergic cells induced by the blockade of CD40-CD154 and CD28-B7 costimulatory pathways can act as potent immunoregulatory cells in vitro, and prolong cardiac allograft survival after adoptive transfer in the presence of rapamycin therapy. This procedure might be clinically useful for prolonging allograft survival if optimal protocols are developed.

Animals ; Antibodies, Monoclonal ; pharmacology ; B7-1 Antigen ; physiology ; CD28 Antigens ; physiology ; CD40 Antigens ; physiology ; CD40 Ligand ; physiology ; Graft Survival ; Heart Transplantation ; Lymphocyte Activation ; Lymphocyte Culture Test, Mixed ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C3H ; Mice, Inbred C57BL ; T-Lymphocytes, Regulatory ; immunology ; Transplantation, Homologous

AIMTo study the characteristic pattern of the age-related growth of the human prostate gland.

METHODSThe volume (weight) of the prostate in 1,601 males, aged from newborn to 92 years, was determined by B-ultrasonography.

RESULTSProstatic volume determination by B-ultrasonography in 1601 males (1301 normal subjects and 300 BPH patients) pointed out that the age-stratified growth of human prostate could be categorized into 4 life stages: (1) the first slow growing phase (from newborn to 9 years): the prostate grows slowly at a rate of 0.14 g per year; (2) the first rapid growing phase (from 10 to 30 years): the prostate grows at a rate of 0.84 g per year; (3) the second slow growing phase (from 30 to 50 years), the prostate grows at a rate of 0.21 g per year; (4) the second rapid growing phase (from 50 to 90 years): the prostate grows at one of the following rates: in one group the growth rate is of 0.50 g per year and in the other 1.20 g per year, leading to benign prostatic hyperplasia (BPH).

CONCLUSIONThe volumes of the prostate are different in different age groups and it grows with age at different rates in four life phases. The prostate growth in phases can be expressed by the following equation: Y=19.36+1.36X'-0.58X'(2+0.33X'3), where Y = prostate volume, X = age (up to 70 years), X'=(X-35.5)/10.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Aging ; Child ; Child, Preschool ; Humans ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Organ Size ; Prostate ; anatomy & histology ; diagnostic imaging ; growth & development ; Ultrasonography

OBJECTIVETo discuss the resection of tumors, reconstruction of defects and the postoperative complications.

METHODSThirty-one patients with tumors around acetabular were treated surgically in People's Hospital between July 1997 and July 2003. The series comprised 19 males and 12 females. Twelve patients were diagnosed with chondrosarcoma, 1 with Ewing sarcoma, 3 with osteosarcoma, 1 with lymphoma, 1 with carcinosarcoma, 1 with malignant fibrohistiocytoma (MFH), 2 with myeloma, 9 with giant cell tumor (GCT), 1 with aneurysmal bone cyst. Among 31 patients with peri-acetabular tumors, 8 were reconstructed with hemi-pelvic prosthesis, 7 with saddle prosthesis, 6 with cauterized tumor bone and total hip arthroplasty, 10 with total hip replacement after curettage of lesion and cemented.

RESULTSAmong 21 patients who underwent tumor resection and reconstruction in region II, 6 had local relapse. Two of 3 patients with osteosarcoma were dead. Five of 12 patients with chondrosarcoma were free of disease. Twenty-one patients with acetabular reconstruction after resection of lesions in region II could sit and stand normally and walked with a cane, several of which even had normal gait.

CONCLUSIONAllograft or pelvic prosthesis can be used to reconstruct the acetabulum after resection of tumors. We must pay more attention on the following points in the surgical treatment of periacetabular tumors: (1) Extensively resect tumors as far as possible; (2) Be acquainted with advantages and disadvantages of different reconstructive methods of acetabulum to prevent the complications; (3) The reconstructed acetabulum is unstable, so the patients must stand with a cane to protect the reconstructed hip joint; (4) Prevent wound necrosis and infection; (5) Surgical treatment of pelvic tumors would easily result in poor wound healing especially in the patients receiving chemotherapy or radiotherapy because of extensive soft tissue stripping. The destroyed soft tissue caused by chemotherapy or radiotherapy may increase the great tissue tension after implantation of allograft. And the factors of poor blood supply and hematoma in the wound theoretically increase the chance of infection.

Acetabulum ; surgery ; Adolescent ; Adult ; Aged ; Arthroplasty, Replacement, Hip ; methods ; Child ; Female ; Hemipelvectomy ; methods ; Humans ; Male ; Middle Aged ; Pelvic Neoplasms ; surgery ; Postoperative Complications ; prevention & control ; Retrospective Studies