Imageable Brachytherapy with Chelator-Free Radiolabeling Hydrogel.

Brachytherapy stands as an essential clinical approach for combating locally advanced tumors. Here, an injectable brachytherapy hydrogel is developed for the treatment of both local and metastatic tumor. Fe-tannins nanoparticles are efficiently and stably radiolabeled with clinical used therapeutic radionuclides (such as 131I, 90Y, 177Lu, and 225Ac) without a chelator, and then chemically cross-linked with 4-armPEG-SH to form brachytherapy hydrogel. Upon intratumoral administration, magnetic resonance imaging (MRI) signal from ferric ions embedded within the hydrogel directly correlates with the retention dosage of radionuclides, which can real-time monitor radionuclides emitting short-range rays in vivo without penetration limitation during brachytherapy. The hydrogel's design ensures the long-term tumor retention of therapeutic radionuclides, leading to the effective eradication of local tumor. Furthermore, the radiolabeled hydrogel is integrated with an adjuvant to synergize with immune checkpoint blocking therapy, thereby activating potent anti-tumor immune responses and inhibiting metastatic tumor growth. Therefore, this work presents an imageable brachytherapy hydrogel for real-time monitoring therapeutic process, and expands the indications of brachytherapy from treatment of localized tumors to metastatic tumors.

Research on the Formation Mechanism of MgO and Al2O3 on Composite Calcium Ferrite Based on DA-RBF Neural Network.

Calcium complex ferrate is an ideal binder phase in the sintered ore phase, and a detailed study of the whole process of calcium complex ferrate generation is of great significance to improve the quality of sintered ore. In this paper, we first investigated calcium ferrate containing aluminum (CFA), which is an important precursor compound for the generation of complex calcium ferrate (SFCA), followed by a series of composite calcium ferrate generation process phase XRD detections and data preprocessing of data. Data correlation and data fitting analysis were combined with composite calcium ferrite phase diagram energy spectrum analysis to obtain the effect of MgO and Al2O3 on the formation of composite calcium ferrite. Then a modified RBF neural network model using the resource allocation network algorithm (RAN) was used to predict the generation trend of complex calcium ferrate. The parameters of the neural network are optimized with the Dragonfly algorithm, compared with the traditional RBF neural network. The prediction accuracy of the improved algorithm was found to be higher, with a prediction result of 97.6%. Finally, the predicted results were based on comparative metallurgical experimental results and data analysis. The validity and accuracy of the findings in this paper were verified.

Teratogenic Toxicity Evaluation of Bladder Cancer-Specific Oncolytic Adenovirus on Mice.

BACKGROUND: In our previous studies, we had demonstrated the efficiency and specificity of constructed bladder tissue-specific adenovirus Ad-PSCAE-UPII-E1A-AR (APU-EIA-AR) on bladder cancer. The virus biodistribution and body toxicity in nude mice have also been investigated. However, the safety of the bladder cancer-specific oncolytic adenovirus on fetal mice and F1 mice should be under intense investigation. OBJECTIVE: In order to evaluate the teratogenic toxicity of bladder cancer-specific oncolytic adenovirus APU-EIA-AR on mice, in this study, we investigated the fetal mice weight, fetal body length and tail length, fetal skeleton development, as well as the F1 mice weight, growth curve, and major organ pathology. These teratogenic toxicity data of bladder tissue-specific adenovirus Ad-PSCAE- UPII-E1A-AR (AD) would provide safe information prior to embarking on clinical trials. METHODS: On the sixth day of being fertilized, the pregnant mice began to be intramuscularly administrated with AD (1×107VP, 1×108VP, 1×109VP) every other day for ten days. The pregnant mice were then divided into two groups. One group was euthanized on the seventeenth day; the fetal mice were taken out, and the bone structure of the infants was observed. The other group was observed until natural childbirth. The Filial Generation (F1) is fed for 30 days; the variations in the growth progress and development were assessed. The mice were then euthanized; The tissues from major organs were harvested and observed under the microscope. RESULTS: In the process of teratogenic toxicity test, the Placenta weight, fetal mice weight, body length, and a tail length of mice fetal in adenovirus treated group did not reveal any alteration. Meanwhile, comparing with the PBS group, there is no obvious change in the skeleton of fetal mice treated with adenovirus. During the development process of F1 mice treated with adenovirus, the changes in mice weight show statistical significance. However, in the progress of the growth curve, this difference is not very obvious. Furthermore, the pathological section showed no obvious alteration in major organs. CONCLUSION: Our study demonstrated that bladder cancer-specific adenovirus Ad-PSCAE-UPII- E1A-AR appears safe in pregnant mice without any discernable effects on fetal mice and F1 development. Hence, it is relatively safe for tumor gene therapy.

Correction: A photo-inducible protein-inorganic nanoparticle assembly for active targeted tumour theranostics.

Correction for 'A photo-inducible protein-inorganic nanoparticle assembly for active targeted tumour theranostics' by Jinbing Xie, Gang Han et al., Nanoscale, 2019, 11, 6136-6144.

A photo-inducible protein-inorganic nanoparticle assembly for active targeted tumour theranostics.

The assembly of protein-inorganic nanoparticles is an important yet challenging approach that is utilized to develop functional materials in numerous areas, such as bio-catalysis, drug delivery, and biosensing. In this study, we report on a facile, photo-inducible self-assembly method to generate protein-inorganic hybrid nanoplatforms. More specifically, photo-treated disulfide bond rich proteins of lysozyme (LYS) were able to be used as host materials in order to encapsulate nanoparticles (i.e., as-synthesized hydrophobic NIR quantum dots (QDs)) and anti-cancer small molecule drugs (i.e., paclitaxel (PTX)), constructing functional theranostic protein-inorganic hybrid nanoparticles. The modification of the functional polymer of cRGD-PEG contributes to the active tumour targeting characteristic of this protein-inorganic nanocarrier. This novel PTX loaded protein-inorganic hybrid nanoplatform showed high tumour homing accumulation as well as effective tumour inhibition. We believe that this general approach represents a new direction for the development of a photo-induced assembly of protein-inorganic nanoparticles towards versatile applications in both materials science and biomedical fields.

Role of 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) in ß-lactoglobulin aggregation.

Protein aggregation is a prevalent phenomenon. It is important to study protein aggregation under different solution conditions. In this study, using 1-anilinonaphthalene-8-sulfonic acid (ANS) fluorescence spectra, we investigated the critical micelle concentration (CMC) of 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP). We also studied the effects of DOTAP on amyloid aggregation of ß-lactoglobulin using intrinsic fluorescence spectra, circular dichroism spectra, thioflavin T fluorescence, a Congo red binding assay and transmission electron microscopy. We observed that DOTAP had a dual role on ß-lactoglobulin amyloid aggregation. DOTAP inhibited the amyloid aggregation below the CMC, while it had the opposite effect above the CMC. Moreover, the results of transmission electron microscopy showed that spherical aggregates were formed above the CMC. These results led us to conclude that cationic lipids could be used as modulators of protein self-assembly.

Resveratrol Induces the Conversion from Amyloid to Amorphous Aggregation of ß-lactoglobulin>.

BACKGROUND: Protein aggregation is a wide-ranging phenomenon. Protein aggregation mainly includes two types: one is amorphous aggregation, the other is amyloid aggregation. In particular, amyloid aggregation in vivo can cause several fatal diseases. We have investigated the influence of resveratrol on the amyloid aggregation of ß-lactoglobulin. The results demonstrated resveratrol inhibited the amyloid aggregation and enhanced the amorphous aggregation of ß- lactoglobulin. OBJECTIVES: The main objective of this study was to investigate the effects of resveratrol on the amyloid aggregation of ß-lactoglobulin. METHODS: ß-lactoglobulin was incubated at pH 2.0 and 70 °C. ThT fluorescence, Congo red and transmission electron microscopy were used to monitor the formation of amyloid aggregates. In addition, resveratrol was added intoß-lactoglobulin solutions. Intrinsic fluorescence, Circular dichroism and 1-Anilinonaphthalene-8-sulfonic Acid (ANS) fluorescence were used to investigate conformational and hydrophobic changes. Furthermore, we also studied the effect of resveratrol on the amyloid aggregation of ß-lactoglobulin by using ThT fluorescence, Congo red and transmission electron microscopy at acidic pH and high temperature. RESULTS: ThT fluorescence, Congo red and transmission electron microscopy analysis showed that ß-lactoglobulin could form the amyloid fibrils when it was incubated under acidic pH and high temperature conditions. At the same time, the analysis also demonstrated resveratrol inhibited the formation of amyloid aggregates and enhanced the formation amorphous aggregates. Intrinsic fluorescence, Circular dichroism and 1-Anilinonaphthalene-8-sulfonic Acid (ANS) fluorescence analysis indicated that resveratrol could alter the conformation and increased the hydrophobicity of ß- lactoglobulin. CONCLUSION: Our results indicated that resveratrol could effectively inhibit the formation of amyloid aggregates and enhance the formation of amorphous aggregates of ß-lactoglobulin. Thus, resveratrol could be a potential inhibitor for preventing the formation of amyloid aggregates.

Investigating the inhibitory effects of zinc ions on amyloid fibril formation of hen egg-white lysozyme.

The amyloid fibrils derived from protein and peptide self-assembly have been studied in many diseases. In the present study, in combination with Thioflavin T(ThT) assay, Congo red(CR),transmission electron microscopy and cell cytotoxicity assay, we investigated the influence of zinc ions on amyloid fibril formation using hen egg white lysozyme (HEWL) as a model protein under high temperature and acidic pH conditions. We observed that HEWL tended to form the amyloid fibrils at pH 2.0 and 60°C, which was consistent with the previous studies. However, as the concentrations of zinc ions increased, the amounts of amyloid fibrils of HWEL gradually reduced, but the overall morphology of individual amyloid fibril was not significantly altered whether or not zinc ions were present. Moreover, by using circular dichroism (CD), ANS and intrinsic fluorescence spectra, we illustrated that zinc ions inhibited the formation of ß-sheet and exposure of hydrophobic regions of HEWL. This work would help to understand the molecular mechanism of amyloid fibril formation.

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.

Potential Mechanisms Responsible for the Antinephrolithic Effects of an Aqueous Extract of Fructus Aurantii.

The potential effects of Fa extract on the prevention and treatment of CaOx nephrolithiasis were analyzed in an ethylene glycol- (EG-) induced CaOx crystallization model in rats and an in vitro assay. Multiple biochemical variables were measured in the urine and kidney. Kidney sections were subjected to histopathological and immunohistochemical analyses. Urolithiasis-related osteopontin (OPN) was evaluated by Western blotting. The in vitro assay revealed the significant inhibition of crystal formation (3.50 ± 1.43) and dilution of formed crystals (12.20 ± 3.35) in the group treated with 1 mg/mL Fa extract compared with the control group (52.30 ± 4.71 and 53.00 ± 4.54, resp.) (p < 0.05). The in vivo experiments showed that prophylactic treatment with Fa aqueous extract significantly prevented EG-induced renal crystallization and pathological alterations compared with nephrolithic rats (p < 0.05). Significantly lower levels of oxidative stress, oxalate, and OPN expression as well as increased citrate and urine output levels were observed in both the low- and high-dose prophylactic groups (p < 0.05). However, in the low- and high-dose therapeutic groups, none of these indexes were significantly improved (p > 0.05) except for urinary oxalate in the high-dose therapeutic groups (p < 0.05). Fa extract prevented CaOx crystallization and promoted crystal dissolution in vitro. Additionally, it was efficacious in preventing the formation of CaOx nephrolithiasis in rats.

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.

Inhibitory effects of ß-ionone on amyloid fibril formation of ß-lactoglobulin.

ß-Lactoglobulin (ß-LG) is the major constituent of whey food, which has been shown to interact with a wide range of aroma compounds. In the present work, a model aroma compound, ß-ionone, is used to investigate the influence of aroma compounds on the urea-induced unfolding of ß-LG at pH 7.0. ß-Ionone is observed to enhance the stability of ß-LG at pH 7.0. Moreover, the amyloid fibrils are observed when ß-LG at pH 7.0 is incubated for 12-20 days at 37 °C in the presence of 3-5M urea. However, the formation of amyloid fibrils is inhibited when ß-ionone is added into the samples and the inhibitory effects follow a concentration-dependent fashion. There is a clear correlation between Cm and lag time. The correlation demonstrates that protein stability affects the amyloid fibril formation of ß-LG. The results highlight the critical role of protein stability and provide an approach to prevent the formation of amyloid fibrils in vitro.

Macromolecular crowding modulates the kinetics and morphology of amyloid self-assembly by ß-lactoglobulin.

The formation of amyloid fibrils by ß-lactoglobulin in the presence of GndHCl has been monitored by using thioflavin T (ThT) fluorescence, Congo Red and transmission electron microscopy (TEM). Large quantities of aggregated protein are formed by incubating ß-lactoglobulin in 2M GndHCl at room temperature and pH 7.0 for about 20 days. The kinetics of fibrillation process can be described by the lag time for formation of stable nuclei (nucleation) and the apparent rate constant for the growth of fibrils (elongation). Moreover, the effects of macromolecular crowding agents, Dextran 70 and polyethylene glycols (PEG), on the amyloid formation of ß-lactoglobulin at pH 7.0 are studied. The results show that the increase in macromolecular crowding agent concentrations results in shorter lag time and faster growth of fibrils. It proves that macromolecular crowding can effectively accelerate the fibril formation of ß-lactoglobulin at neutral pH. At the same time, it can be observed that the amplitude of the ThT fluorescence intensity decreases as the Dextran 70 concentration is increased. The observation suggests that the yield of amyloid fibrils is significantly reduced by the addition of macromolecular crowding agents. The conclusion is further confirmed by the transmission electron microscopy. In addition, the results of transmission electron microscopy also indicate that macromolecular crowding can alter the fibril morphology of ß-lactoglobulin. In brief, our findings demonstrate that the effects of macromolecular crowding are essential to the understanding of protein amyloid self-assembly occurring in vivo.

Tamsulosin for ureteral stones: a systematic review and meta-analysis of a randomized controlled trial.

BACKGROUND/AIMS: To evaluate the efficacy of tamsulosin as a medical expulsive therapy of ureteral stones. METHODS: We searched PubMed, EMBASE, the Cochrane Library, and ISI-Science Citation Index up to December 2011. All randomized controlled trials were identified in which patients were randomized to receive either tamsulosin or standard therapy with/without placebo for ureteral stones. Outcome measures assessed were overall stone expulsion rate (primary) and expulsion time, and the number of pain episodes (secondary). Three authors independently assessed study quality and extracted data. All data were analyzed using RevMan 5.0. RESULTS: Twenty-nine trials with a total of 2,763 patients met the inclusion criteria. The pooled analysis showed a 19% improvement in stone clearance with tamsulosin. According to the doses of tamsulosin, the pooling effects of tamsulosin were analyzed, with a higher expulsion rate obtained than in controls. Compared with calcium channel blockers, there was a higher stone expulsion rate in tamsulosin. In addition, a shorter expulsion time, fewer colic episodes and adverse effects were observed. CONCLUSIONS: Tamsulosin is a safe and effective medical expulsive therapy choice for ureteral stones. It should be recommended for most patients with distal ureteral stones before stones are 10 mm in size. In future, high-quality multicenter, randomized and placebo-controlled trials are needed to evaluate the outcome.

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.

Growth inhibitory effects of quercetin on bladder cancer cell.

Quercetin, a flavonoid found in many fruits and vegetables, belongs to an extensive class of polyphenolic compounds. Previous studies reported that quercetin inhibits the proliferation of various cancer cells and tumor growth in animal models. We investigated the growth inhibition and colony formation of quercetin on three bladder cancer cells (EJ, J82 and T24). The expression of tumor suppressor genes and oncogenes such as P53, Survivin, PTEN, as well as the methylation status of these genes was also evaluated. We observed that quercetin induced apoptosis in bladder cancer cells in a time- and dose-dependent manner. Quercetin (100 micromolars) significantly inhibited EJ, T24 and J82 cell growth accompanied by an increase in the G0/G1 phase. In all cell lines, quercetin decreased the expression of mutant P53 and Survivin proteins. However, there was no change in the level of PTEN protein. Moreover, the DNA methylation levels of the estrogen receptor (Er-beta), P16INK4a and RASSF1A were strongly decreased (from 35 to 70%) in the quercetin-treated group compared to the control. In conclusion, our study suggested that quercetin inhibits growth, colony formation and hypermethylation of bladder cancer cell lines. Quercetin-induced apoptosis might be associated with a decrease in mutant P53 and Survivin proteins.

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.