Membrane fluidity is involved in the regulation of heat stress induced secondary metabolism in Ganoderma lucidum.

Ganoderma lucidum has become a potential model system for evaluating how environmental factors regulate the secondary metabolism of basidiomycetes. Heat stress (HS) is one of the most important environmental factors. It was previously reported that HS could induce the biosynthesis of ganoderic acids (GA). In this study, we found that HS increased GA biosynthesis and also significantly increased cell membrane fluidity. Furthermore, our results showed that addition of the membrane rigidifier dimethylsulfoxide (DMSO) could revert the increased GA biosynthesis elicited by HS. These results indicate that an increase in membrane fluidity is associated with HS-induced GA biosynthesis. Further evidence showed that the GA content was decreased in D9des-silenced strains and could be reverted to WT levels by addition of the membrane fluidizer benzyl alcohol (BA). In contrast, GA content was increased in D9des-overexpression strains and could be reverted to WT levels by the addition of DMSO. Furthermore, both membrane fluidity and GA biosynthesis induced by HS could be reverted by DMSO in WT and D9des-silenced strains. To the best of our knowledge, this is the first report demonstrating that membrane fluidity is involved in the regulation of heat stress induced secondary metabolism in filamentous fungi.

Transrectal microwave thermotherapy causing a short-time influence on sperm quality in Chinese chronic nonbacterial prostatitis patients.

Chronic prostatitis can affect the sperm's quality. Previous studies have shown that transrectal microwave thermotherapy (TRMT) results in symptomatic relief in patients with chronic prostatitis, but the effects on sperm have not been carefully investigated. This study evaluates the impact of TRMT on the relief or decrease of symptoms and quality of sperm when used to treat patients with chronic nonbacterial prostatitis. Sixty patients were enrolled in the study. TRMT treatment was administered over 5 days, 1 h per day. Semen examination was carried out pretreatment and immediately at the conclusion of the 5-day treatment. Also, it was repeated 1 month, 3 months, and 6 months later. The treatment's symptom relief efficacy was evaluated using the National Institutes of Health Chronic Prostatitis Symptom Index (NIH-CPSI). After the treatment, the overall NIH-CPSI scores were lower compared to those of pretreatment. In addition, the white blood cells and lecithin in expressed prostatic secretion were normal after the treatment. The sperm count was decreased by 23.8% 3 months after the treatment, sperm motility was reduced by 10.3% immediately after treatment, and sperm deformity was increased by 17.2%. The sperm volume and PH were not affected. However, the sperm quality recovered after treatment and the malformation rate was also lower at 6 months after treatment. TRMT is a favorable and safe treatment option for patients with nonbacterial chronic prostatitis. It could relieve the patient's symptoms and impact on sperm quality in the short-term.

[Correlation among serum MBL, MASP-2, HsCRP and C3 levels in rheumatoid arthritis].

OBJECTIVE: To investigate the correlation among serum levels of manning-binding lectin (MBL), MBL-associated serine proteases-2 (MASP-2), complement C3 and high-sensitive C reactive protein (HsCRP) in patients with rheumatoid arthritis (RA). METHODS: Fasting venous blood were collected from 50 RA patients (25 in active stage and 25 in remission) and 40 healthy subjects for detecting serum levels of MBL, MASP-2, complement C3 and HsCRP using enzyme-linked immunosorbent assay (ELISA) and immune turbidity assay. RESULTS: The serum levels of MBL and MASP-2 were significantly lower and HsCRP level was significantly higher in patients with RA (in both acute stage and remission) than in the healthy control group (P<0.05), but complement C3 level was similar between the RA patients and control group. Bivariate Pearson correlation analysis showed that in RA patients, MBL was positively correlated with MASP-2 level (r=0.550, P=0.001) and negatively with HsCRP (r=-0.323, P=0.022) but not correlated with C3 (r=-0.022, P=0.882); MASP-2 was negatively correlated with HsCRP (r=0.453, P=0.453) and was not correlated with C3 (r=0.049, P=0.738). ROC curve analysis revealed the largest area under curve (AUC) of HsCRP (0.844, P=0.001) and smaller AUCs of MBL (0.025, P=0.001) and MASP-2 (0.266, P=0.001). HsCRP had a much higher sensitivity (84%) than MBL (10%) and MASP-2 (40%) in the diagnosis of RA. CONCLUSION: In RA patients, MBL and MASP-2 are negatively correlated with HsCRP level. Serum MBL and MASP-2 levels decrease with the progression of joint injury in RA patients, suggesting their involvement in the pathological process of RA; but due to their low sensitivity, they are not appropriate indicators for evaluating the disease activity of RA.

Subcutaneous Injections of the Mannose-Sensitive Hemagglutination Pilus Strain of Pseudomonas aeruginosa Stimulate Host Immunity, Reduce Bladder Cancer Size and Improve Tumor Survival in Mice.

We wished to evaluate the effects of Pseudomonas aeruginosa (mannose-sensitive hemagglutination pilus strain, PA-MSHA) as an immunostimulating and anti-tumor agent for treatment of bladder cancer. Immunostimulating effects were assessed by the in vitro proliferation assay of murine splenic lymphocytes. Anti-tumor effects were studied in a subcutaneous tumor model established in female C57BL/6 mice using the MB49 bladder cell line. These mice received subcutaneous injections of normal saline (control group) or PA-MSHA (high, medium, or low dose, respectively, 1.6-2.0 × 10(9), 3.2- .0 × 10(8), 6.4-8.0 × 10(7) CFU/ml) twice a week for 3 weeks. Mice survival, tumor volume, vascular endothelial growth factor (VEGF) expression, microvessel density (MVD), serum levels of TNF-α and IFN-γ, and blood CD4(+) /CD8(+) counts were the study outcomes. We observed that PA-MSHA promoted the growth of splenic lymphocytes in vitro. In the murine tumor model, PA-MSHA prolonged mice survival and reduced tumor growth. Furthermore, VEGF and MVD were also diminished by PA-MSHA. Mice that received high and medium dose of PA-MSHA had significantly higher serum levels of IFN-γ and TNF-α (days 21 and 28), and higher levels of CD4(+) /CD8(+) cells (days 21 and 28). In conclusion, PA-MSHA exerts beneficial effects on increasing proliferation of murine splenic lymphocytes in vitro and inhibits the growth of bladder tumor in a murine model. Therefore, PA-MSHA may be useful an immunostimulating and anti-tumor agent for bladder cancer therapy.

Detection of nuclear matrix protein 22 and survivin baseline level in patients after radical cystectomy.

We investigated the baseline levels of urine nuclear matrix protein 22 (U-NMP22) and survivin in urine after radical cystectomy for primary invasive bladder cancer. We measured U-NMP22 and survivin values in 72 patients with four types of urinary diversion (Indiana bladder, Bricker bladder, Mainz bladder and orthotopic bladder) after radical cystectomy and 25 healthy volunteers. We also analyzed the relation between the U-NMP22 and survivin level and other variables among patients with continent urinary diversion and incontinent urinary diversion as well as healthy controls, and found that the U-NMP22 and survivin values were not associated with postoperative interval or gender. The U-NMP22 values (mean ± standard error) for continent urinary diversion, incontinent urinary diversion and healthy controls were 12.08 ± 0.10, 16.62 ± 0.15 and 0.01 ± 0.00 U/ml, respectively. The survivin values (mean ± standard error) for continent urinary diversion, incontinent urinary diversion and healthy controls were 0.47 ± 0.06, 0.69 ± 0.16 and 0.02 ± 0.03 U/ml, respectively. The U-NMP22 and survivin values in the Bricker bladder group were significantly higher than the values in the other three groups. We noted that increased levels of U-NMP22 and survivin after radical cystectomy varied according to different predictors, which may be useful for designing strategies to follow these cases.

In vitro enhanced cytotoxicity of tumor-infiltrating lymphocytes transfected with tumor necrosis factor-related apoptosis-inducing ligand and/or interleukin-2 gene in human renal cell carcinoma.

OBJECTIVES: To investigate whether tumor-infiltrating lymphocytes (TILs) transfected with tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and interleukin-2 (IL-2) genes are capable of improving the potency and efficacy of propagation and cytotoxicity against renal cell carcinoma (RCC) cells in vitro. METHODS: A mammal expression vector system was constructed. TILs were transfected by liposome-mediated gene transfection. The degree of cytokine mRNA expression was evaluated with Northern blot. Protein expression was determined with Western blot and enzyme-linked immunosorbent assay. Cytotoxicity of TILs against autologous RCC cells and the human RCC cell line (786-0) were examined by chromium release assay. Flow cytometric analyses were performed to determine the apoptosis of tumor cells. RESULTS: A high level of expression of the human TRAIL and IL-2 stable transfected TILs was observed. The mean IL-2 production was 22.6 +/- 5.2, 507.7 +/- 52.4, and 549.0 +/- 74.0 ng/10(6) cells/24 hours in the TIL/parental, TIL/IL-2, and TIL/TRAIL+IL-2 genes, respectively. The mean cytotoxicity (effector/target ratio 20:1) of TIL/parental, TIL/IL-2, TIL/TRAIL, and TIL/TRAIL+IL-2 against autologous RCC cells in the percentage of cytolysis was 21.2% +/- 4.8%, 32.1% +/- 5.5%, 63.5% +/- 6.6%, and 78.1% +/- 9.63%, respectively. These four groups showed cytotoxic activity against allogeneic 786-0 RCC cells; the corresponding values were 9.8% +/- 3.5%, 12.3% +/- 3.4%, 24.1% +/- 4.9%, and 30.4% +/- 6.2%. The number of apoptotic cells was significantly greater for autologous RCC cells than for 786-0 cells after TIL/TRAIL and TIL/TRAIL+IL-2 treatment. CONCLUSIONS: TIL/TRAIL+IL-2 and TIL/IL-2 were expanded by autocrine IL-2. TIL/TRAIL+IL-2 and TIL/TRAIL showed significant cytotoxicity that was induced by TRAIL. TILs, including parental TILs and transfected TILs, demonstrated a potent cytotoxicity against RCC cells with remarkable selectivity. Autologous RCC cells seemed more sensitive than allogeneic RCC cells.

Urothelial injury to the rabbit bladder caused by calcium dissolving agents including two new citrate solutions.

We compared the urothelial injury to the bladder caused by four agents capable of dissolving calcium salts. The solutions were administrated in an antegrade way through left ureterostomies in 54 rabbits for periods of 24, 48 and 72 h. The bladders were then removed and three routine histological sections were made for each. The following six solutions were used: physiological sodium chloride solution (Phys), artificial urine (Art), 0.03 M disodium EDTA buffered to pH 8.5 with triethanolamine (EDTA), 10% Renacidin (R), test solution 2 (S2, using D-gluconic acid-lactone and other compounds that differ from R in terms of ingredients or quantity), and test solution 1 (S1, using D-gluconic-acid instead of D-gluconic acid-lactone in S2 but keeping the other ingredients the same) for irrigation. At 24 h there was no observable urothelial damage caused by perfusion with Phys or Art; solutions R, S1 and S2 caused approximately the same level of injury to the rabbit bladder mucosa; however, irrigation with disodium-EDTA caused more serious urothelial injury than R, S1 and S2 (P<0.05, chi2-test) and may be unacceptable. The damage to bladder tissues treated with S1 and S2 was less than that caused by R, but this was not significant (P>0.05, chi2-test). Following a prolonged irrigation time, all of these solutions cause further urothelial damage, but EDTA caused the most, followed by R, S1, S2, Phys or Art, respectively, at 48 and 72 h. In view of the better solubility effect of solutions S1 and S2 compared with R, it might be justified in accepting the more pronounced urothelial irritation caused these solutions, but in order to enhance their effectiveness and reduce urothelial injury further study will be needed.

Effects of pollen extract EA-10, P5 on chronic prostatitis or infertility with chronic prostatitis.

AIM: To determine the drug action mechanism of pollen extract EA-10, P5 on the treatment of chronic prostatitis (CP) or infertility with CP. METHODS: Malondiadehyde (MDA), superoxide dismutase (SOD), and nitrogen monoxide (NO) were measured by biochemical assay, and zinc content was assayed by atomical spectrophotography in the pre-treatment and post-treatment of CP or infertility with CP. RESULTS: Compared with control group, leukocytes in expressed protatic secretion (LEPS), MDA, and NO were increased, and zinc content and SOD were decreased significantly in the pre-treatment of CP. After the treatment, LEPS was improved, and MDA and NO were reduced, while zinc content were increased apparently and the alteration of SOD was not evident (P>0.05). In the pre-treatment of infertility with CP, LEPS, MDA, NO, sperm viability, and seminal leukocytes were obviously higher and seminal plasma SOD, zinc content, and sperm motility were obviously lower than those in control group. After the treatment, LEPS, sperm motility, and sperm viability were improved, MDA, NO, and seminal leukocytes were decreased, SOD and zinc content were increased markedly. CONCLUSION: There was inter-correlation between oxygen free radicals (OFR) and occurrence, development, and recovery of CP; Change of OFR may be involved in the drug action mechanism of EA-10, P5 in the treatment of CP or infertility with CP.