Identification of clinical prognostic factors and analysis of ferroptosis-related gene signatures in the bladder cancer immune microenvironment.

BACKGROUND: Bladder cancer (BLCA) is a prevalent malignancy affecting the urinary system and poses a significant burden in terms of both incidence and mortality rates on a global scale. Among all BLCA cases, non-muscle invasive bladder cancer constitutes approximately 75% of the total. In recent years, the concept of ferroptosis, an iron-dependent form of regulated cell death marked by the accumulation of lipid peroxides, has captured the attention of researchers worldwide. Nevertheless, the precise involvement of ferroptosis-related genes (FRGs) in the anti-BLCA response remains inadequately elucidated. METHODS: The integration of BLCA samples from the TCGA and GEO datasets facilitated the quantitative evaluation of FRGs, offering potential insights into their predictive capabilities. Leveraging the wealth of information encompassing mRNAsi, gene mutations, CNV, TMB, and clinical features within these datasets further enriched the analysis, augmenting its robustness and reliability. Through the utilization of Lasso regression, a prediction model was developed, enabling accurate prognostic assessments within the context of BLCA. Additionally, co-expression analysis shed light on the complex relationship between gene expression patterns and FRGs, unraveling their functional relevance and potential implications in BLCA. RESULTS: FRGs exhibited increased expression levels in the high-risk cohort of BLCA patients, even in the absence of other clinical indicators, suggesting their potential as prognostic markers. GSEA revealed enrichment of immunological and tumor-related pathways specifically in the high-risk group. Furthermore, notable differences were observed in immune function and m6a gene expression between the low- and high-risk groups. Several genes, including MYBPH, SOST, SPRR2A, and CRNN, were found to potentially participate in the oncogenic processes underlying BLCA. Additionally, CYP4F8, PDZD3, CRTAC1, and LRTM1 were identified as potential tumor suppressor genes. Significant discrepancies in immunological function and m6a gene expression were observed between the two risk groups, further highlighting the distinct molecular characteristics associated with different prognostic outcomes. Notably, strong correlations were observed among the prognostic model, CNVs, SNPs, and drug sensitivity profiles. CONCLUSIONS: FRGs are associated with the onset and progression of BLCA. A FRGs signature offers a viable alternative to predict BLCA, and these FRGs show a prospective research area for BLCA targeted treatment in the future.

Irisin protected hemopoietic stem cells and improved outcome of severe bone marrow failure.

Acquired aplastic anemia (AA) is a bone marrow failure (BMF) disease, characterized by fatty bone marrow (BM) and BM hypocellularity resulted from auto-immune dysregulated T cells-mediated destruction of BM haemopoietic stem cells (HPSC). The objective of this study was to investigate potential therapeutic effect of irisin, a molecule involved in adipose tissue transition, on AA mouse model. Our results showed that the concentration of irisin in serum was lower in AA patients than in healthy controls, suggesting a role of irisin in the pathogenesis of AA. In the AA mice, irisin administration prolonged the survival rate, prevented or attenuated peripheral pancytopenia, and preserved HPSC in the BM. Moreover, irisin also markedly reduced BM adipogenesis. In vitro results showed that irisin increased both cell proliferation and colony numbers of HPSC. Furthermore, our results demonstrated that irisin upregulated the expression of mitochondrial ATPase Inhibitory Factor 1 (IF1) in HPSC, inhibited the activation of mitochondrial fission protein (DRP1) and enhanced aerobic glycolysis. Taken together, our findings indicate novel roles of irisin in the pathogenesis of AA, and in the protection of HPSC through stimulation of proliferation and regulation of mitochondria function, which provides a proof-of-concept for the application of irisin in AA therapy.

Preparation, function, and safety evaluation of a novel degradable dermal filler, the cross-linked poly-γ-glutamic acid hydrogel particles.

Poly-γ-glutamic acid (PGA) is a naturally degradable hydrophilic linear microbial polymer with moisturizing, immunogenic, cross-linking, and hydrogel water absorption properties similar to hyaluronic acid, a biomaterial that is commonly used as a dermal filler. To explore the development feasibility of cross-linked PGA as a novel dermal filler, we studied the local skin response to PGA fillers and the effect of various cross-linking preparations on the average longevity of dermal injection. Injection site inflammation and the formation of collagen and elastin were also determined. PGA hydrogel particles prepared using 28% PGA and 10% 1,4-butanediol diglycidyl ether showed optimal filler properties, resistance to moist heat sterilization, and an average filling longevity of 94.7 ± 61.6 days in the dermis of rabbit ears. Local redness and swelling due to filler injection recovered within 14.2 ± 3.6 days. Local tissue necrosis or systemic allergic reactions were not observed, and local collagen formation was promoted. Preliminary results suggested that dermal injection of cross-linked PGA particles appeared safe and effective, suggesting that cross-linked PGA particles could be developed as a new hydrogel dermal filler.

Mesenchymal stem cells significantly improved treatment effects of Linezolid on severe pneumonia in a rabbit model.

The present study aimed to investigate whether co-administration of mesenchymal stromal cells (MSC) and linezolid (LZD) into a rabbit model of methicillin-resistant Staphylococcus aureus (MRSA)-infected pneumonia would bring a synergistic therapeutic effect. Human umbilical cord-derived MSCs (hUMSCs) were isolated and characterized. A rabbit model of pneumonia was constructed by delivering 1 × 1010 CFU MRSA via a bronchoscope into the basal segment of lower lobe of right lung. Through analyzing vital sign, pulmonary auscultation, SpO2, chest imaging, bronchoscopic manifestations, pathology, neutrophil percentage, and inflammatory factors, we verified that a rabbit model of MRSA-induced pneumonia was successfully constructed. Individual treatment with LZD (50 mg/kg for two times/day) resulted in improvement of body weight, chest imaging, bronchoscopic manifestations, histological parameters, and IL-10 concentration in plasma (P<0.01), decreasing pulmonary auscultation, and reduction of IL-8, IL-6, CRP, and TNF-α concentrations in plasma (P<0.01) compared with the pneumonia model group at 48 and 168 h. Compared with LZD group, co-administration of hUMSCs (1 × 106/kg for two times at 6 and 72 h after MRSA instillation) and LZD further increased the body weight (P<0.05). The changes we observed from chest imaging, bronchoscopic manifestations and pathology revealed that co-administration of hUMSCs and LZD reduced lung inflammation more significantly than that of LZD group. The plasma levels of IL-8, IL-6, CRP, and TNF-α in combined group decreased dramatically compared with the LZD group (P<0.05). In conclusion, hUMSCs administration significantly improved therapeutic effects of LZD on pneumonia resulted from MRSA infection in a rabbit model.

Analysis of the correlation between lipotoxicity and pituitary-thyroid axis hormone levels in men and male rats.

Lipotoxicity seriously harms human health, but it is unclear whether lipotoxicity is detrimental to the pituitary. We investigated the correlation between serum triglyceride and pituitary axis hormone levels in epidemiological and animal studies. In the epidemiological study, serum thyroid-stimulating hormone (TSH), follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels were greater in male patients with isolated hypertriglyceridemia than in controls, whereas adrenocorticotropin (ACTH) levels were lower in the patients with hypertriglyceridemia. Pituitary hormone levels correlated with triglyceride levels, even after adjustment for potential confounders. In the animal study, male rats were fed a high-fat or control diet for 28 weeks. As the duration of high-fat feeding increased, the serum and pituitary triglyceride concentrations increased. At early times, the high-fat diet elevated serum TSH and triiodothyronine. At later times, much higher serum TSH levels coupled with reduced thyroxine were observed in the high-fat group. Serum levels of pituitary-gonadal and pituitary-adrenal axis hormones were not affected by the diet. The mRNA and protein expression of Tshß were greater in the high-fat group than in the control group, whereas expression of Fshß, Lhß and Acth had no difference between the groups. Overall, serum triglyceride levels were associated with pituitary-thyroid axis hormone levels.

Relationship between gene polymorphisms and prostate cancer risk.

OBJECTIVE: To investigate the relationship between genetic factor and prostate cancer (Pca) risk and the possible cause in it. METHODS: The polymorphisms of cytochrome P450 family 17 (CYPl7) rs743572, p27 V109G and androgen receptor (AR) gene CAG repeat length in peripheral blood from 70 cases and 70 controls were detected through the polymerase chain reaction-restriction fragment length polymorphism technique or short tandem repeat-polymerase chain reaction technique. Then, according to the results of case-control study, the recombinant plasmids containing the wild/mutant p27 gene were constructed and transfected Pca LNcap cells. After 24 and 72 h of transfection, the cell proliferative activity was determined by MTT method, cell cycle distribution and apoptosis was detected by flow cytometry, and the expression level of bcl-2, caspase-3 and p27 protein was determined by Western-blot. RESULTS: In three target polymorphisms, only p27 V109G polymorphism was related to Pca risk (P = 0.030, OR = 0.202, 95% CI = 0.042-0.973). Pca risk of p27-109G allele was lower than -109V allele (P = 0.006, OR = 0.285, 95% CI = 0.110-0.737). Cells transfected with wild/mutant p27 gene both showed the higher cells apoptosis rate and the lower cell proliferative activity than mock cells (P < 0.05 or 0.01), the regulatory effect of mutant p27 on cell proliferation and apoptosis was stronger than the wild p27 (P < 0.05). CONCLUSIONS: p27-109G allele that could cause higher p27 protein expression than -109V allele in LNcap cells, maybe is the protective factor of Pca.

Effects of different interchain linkers on biological activity of an anti-prostate cancer single-chain bispecific antibody.

BACKGROUND: A single-chain bispecific antibody (scBsAb; an engineered antibody), has promising clinical applications. Nonetheless, the effect of different interchain linkers on its activity is poorly understood. METHODS: Gene synthesis was used to splice the anti-γ-seminoprotein single-chain antibody (anti-γ-Sm scFv) gene with the anti-CD3 single-chain antibody (anti-CD3 scFv) gene via different interchain peptide linkers. The Phyre2 software was used to predict spatial configuration of different scBsAbs. Eukaryotic expression vectors carrying scBsAbs were constructed by molecular cloning techniques and these plasmids were transfected into HeLa cells with liposomes. scBsAbs were purified by Ni(2+)-NTA agarose and analysed for antigen binding by an enzyme-linked immunosorbent assay (ELISA). Blood pharmacokinetics and inhibition of prostate tumour growth in nude mice were analysed in in vivo experiments. RESULTS: Bioinformatics analysis and prediction showed that none of the three linkers, Fc, 205C', and HSA, had a significant effect on protein folding of anti-γ-Sm scFv or anti-CD3 scFv. Nevertheless, the spatial structures of the three linkers were noticeably different. Anti-γ-Sm × anti-CD3 scBsAb with an Fc, 205C', or HSA linker was successfully constructed, and these antibodies had similar protein expression levels. ELISA showed that all the three scBsAbs bound to Jurkat cells and the LNCaP membrane antigen, although binding of (205C')scBsAb was weaker than that of the two parental scFvs (P < 0.05). In contrast, binding strength of (HSA)scBsAb and (Fc)scBsAb was close to that of the parental scFvs (P > 0.05). Pharmacokinetic analysis showed that the half-clearance time of the elimination phase (T(1/2ß)) for (HSA)scBsAb was the longest: up to 4.4 h. Compared with γ-Sm ScFv, the three scBsAbs all had a much stronger inhibitory effect on the growth of prostate cancer (P < 0.05), but there were no significant differences among the three scBsAbs (P > 0.05). CONCLUSIONS: HSA is the optimal linker for the anti-γ-Sm × anti-CD3 scBsAb and may improve antigen-binding affinity of antibodies and prolong physiological retention time. Interchain linkers affect the function of scBsAbs; these effects may have important implications for construction of antibodies.

[Clinical significance of serum telomerase mRNA in patients with prostate cancer].

OBJECTIVE: To explore the clinical significance of detecting serum mRNA of telomerase in the diagnosis of prostate cancer. METHODS: Serum samples were collected from 29 patients with prostate cancer and 29 age-matched patients with cardiovascular or metabolic disease as non-tumor controls. Sera samples from 15 healthy age-matched subjects were used as healthy control. Detection of serum telomerase mRNA was performed with real-time reverse transcriptase quantitative polymerase chain reaction (PCR). RESULTS: Serum telomerase mRNA was detectable in 89.7% (26/29) patients with prostate cancer, but almost undetectable in non-tumor (6.9%, 2/29) and healthy control groups(1/14). CONCLUSION: Using real-time PCR for detecting serum telomerase mRNA may be an auxiliary method for diagnosing and monitoring of prostate cancer.

[Expression and clinical significance of transcription factors in prostate cancer].

OBJECTIVE: To explore the roles of various transcription factors in the occurrence and development of prostate cancer. METHODS: A total of 139 specimens with prostate cancer (PCa) and 83 specimens with benign prostatic hyperplasia (BPH) from hospitalized patients at our hospital from 2008 to 2011 were enrolled. The mRNA expressions of c-Myc, Klf4, Nanog, Oct4A and Sox2 were determined by quantitative real-time polymerase chain reaction (PCR) and the expressions of Klf4 isoforms by conventional PCR. Immunohistochemical method was used for the detection of Klf4 protein via tissue microarray in 404 prostate samples. RESULTS: No significant difference existed in the expressions of Nanog, Oct4A and Sox2 genes between BPH and PCa samples. And the expressions of c-Myc and Klf4 genes were significantly higher in PCa than those in BPH specimens. Immunohistochemical results showed that Klf4 protein could be detected in a large majority of epithelial prostatic cells irrespective of malignant transformation. However, it was predominantly located cytoplasmically in PCa tissues and remained consistent with the expression of a differentially spliced Klf4α isoform. CONCLUSION: Klf4 is highly expressed in both BPH and PCa tissues. But in malignant cells, a specific gene product Klf4α is predominantly detectable in cytoplasm. The positioning of Klf4 protein may have an important relationship with its role in the tumorigenic process.

TRAP1 regulates proliferation, mitochondrial function, and has prognostic significance in NSCLC.

UNLABELLED: The TNF receptor-associated protein 1 (TRAP1) is a mitochondrial HSP that has been related to drug resistance and protection from apoptosis in colorectal and prostate cancer. Here, the effect of TRAP1 ablation on cell proliferation, survival, apoptosis, and mitochondrial function was determined in non-small cell lung cancer (NSCLC). In addition, the prognostic value of TRAP1 was evaluated in patients with NSCLC. These results demonstrate that TRAP1 knockdown reduces cell growth and clonogenic cell survival. Moreover, TRAP1 downregulation impairs mitochondrial functions such as ATP production and mitochondrial membrane potential as measured by TMRM (tetramethylrhodamine methylester) uptake, but it does not affect mitochondrial density or mitochondrial morphology. The effect of TRAP1 silencing on apoptosis, analyzed by flow cytometry and immunoblot expression (cleaved PARP, caspase-9, and caspase-3) was cell line and context dependent. Finally, the prognostic potential of TRAP1 expression in NSCLC was ascertained via immunohistochemical analysis which revealed that high TRAP1 expression was associated with increased risk of disease recurrence (univariate analysis, P = 0.008; multivariate analysis, HR: 2.554; 95% confidence interval, 1.085-6.012; P = 0.03). In conclusion, these results demonstrate that TRAP1 impacts the viability of NSCLC cells, and that its expression is prognostic in NSCLC. IMPLICATIONS: TRAP1 controls NSCLC proliferation, apoptosis, and mitochondrial function, and its status has prognostic potential in NSCLC.

[Expression of GDNF in rat Sertoli cells cultured in vitro at different temperatures].

OBJECTIVE: To explore the mechanism of hyperthermia inducing infertility by observing the expression of glial cell line-derived neurotrophic factor (GDNF) in rat Sertoli cells cultured in vitro at different temperatures. METHODS: Using combination enzyme digestion and selective adhesion, we isolated Sertoli cells from male Wistar rats and cultured them in vitro at different temperatures, followed by observation of the changes in their adhesion and morphology and identification by FasL immunohistochemical staining. We divided the Sertoli cells into a control group (35 degrees C) and four experimental groups (36 degrees C, 37 degrees C, 38 degrees C, and 39 degrees C), measured their proliferation by CCK-8, observed their morphology and structure by HE staining, and determined the expression of GDNF by RT-PCR, immunofluorescence and Western blot. RESULTS: Sertoli cells were successfully isolated and in vitro-cultured, with a purity of (95.30 +/- 2.15)% (n = 10). The CCK-8 assay showed that the proliferation of the Sertoli cells was the highest at 36 degrees C, gradually decreasing with the temperature above 36 degrees C, and significantly inhibited at 39 degrees C (P < 0.01). Immunofluorescence revealed the expression of GDNF in the cytoplasm, with the highest fluorescence intensity at 36 degrees C. RT-PCR and Western blot exhibited a decreasing trend of the GDNF expression with the increasing temperature above 36 degrees C. There were statistically significant differences in the expression of GDNF between the control group and the four experimental groups (P < 0.01). CONCLUSION: The proliferation and GDNF expression of in vitro-cultured Sertoli cells differ significantly at different temperatures. At > 36 degrees C, the higher the temperature is, the lower the Sertoli cell proliferation and GDNF expression are. Our findings suggest that high temperature above 36 degrees C suppresses the function of Sertoli cells and may also damage spermatogenesis.

Establishment and identification of induced pluripotent stem cells in liver cancer patients.

OBJECTIVE: To induce pluripotent stem (IPS) cells from fibrocytes that are separated from liver cancer patients. METHODS: The fibrocytes were reprogrammed to IPS cells by lentiviral vector, stained and identified by immunohistochemistry. RESULTS: The IPS cells were successfully established from fibrocytes after infection, and IPS cell clones formed in round shape under a microscopy. The induction rate was 0.013%±0.007%. No tumor formed at the back of nude mice within 8 weeks after the inoculation of cell clones. However, tetatoma appeared in nude mice within 1 week after IPS inoculation. A few tumors formed in nude mice within 4 weeks after the inoculation of cell clones. However, subcutaneous tumors formed within 1 week after IPS inoculation. The induced IPS cells showed three germ layers in tetatoma. Nanog and OCT4 in the induced IPS cells showed hypomethylation. SSEA-A, TRA-1-6-, TRA-1-81 and Nanog were highly expressed in the induced IPS cells, indicating the IPS cells possessed the similar ability as the stem cells. CONCLUSIONS: The IPS cells of liver cancer patients can be established effectively from fibrocytes and can be cultured stably in vitro, which provides an approach for the treatment of intermediate or advanced stage liver cancer.

[Hypoxia reduces the proliferation and occludin expression of primary sertoli cells].

OBJECTIVE: To investigate the effect of hypoxia on the proliferation and occludin expression of primary rat Sertoli METHODS: We constructed a primary Sertoli cell system by two-step enzymatic digestion in 18 -22 days old Wistar rats and identified it by oil red O and immunofluorescence methods. We randomly divided the Sertoli cells into five groups to be cultured in oxygen at the concentrations of 20%, 15%, 10%, 5% and 1%, respectively, for 6, 12, 24, 48 and 72 hours. We detected the proliferation of the Sertoli cells by CCK-8 assay, determined the expression of occludin by Western blot, and analyzed the differences among the five groups. RESULTS: Oil red O staining revealed red lipid droplets in the cytoplasm of the Sertoli cells, and immunofluorescence showed the positive expression of the FasL protein, with the purity of Sertoli cells over 95% in vitro. Compared with the 20% normoxic group, the proliferation of the Sertoli cells was gradually reduced in the 15% and 10% hypoxia groups, and significantly declined in the 5% and 1% groups (P < 0.01). At 12 hours, the expression of occludin began to decrease with the prolonging of time and reduction of oxygen concentration (P < 0.01). CONCLUSION: Hypoxia suppresses the proliferation of Sertoli cells and reduces the expression of occludin. It could be inferred that hypoxia could damage the integrity of blood-testis barrier and spermatogenesis of the testis.

[High temperature reduces the proliferation of and occludin expression in rat Sertoli cells in vitro].

OBJECTIVE: To explore the action mechanisms of temperature in male infertility or subfertility by observing the effects of different temperatures on the proliferation of and occludin (OCLN) expression in rat Sertoli cells in vitro. METHODS: We isolated Sertoli cells from the testis of male Wistar rats, and performed oil red O staining and immunohistochemistry to identify their FasL. We cultured the Sertoli cells at 34 degrees C (control group) and at 35, 36, 37, 38 and 39 degrees C (experimental groups) for 4 days. Then we measured their proliferation by CCK-8 assay, observed their morphology and structure by hematoxylin-eosin staining, and determined their OCLN expression level by Western blotting and immunofluorescence. RESULTS: The purity of the isolated Sertoli cells was (96.20 +/- 1.95)%. CCK-8 assay indicated that the proliferation of the Sertoli cells was increased between 34 and 36 degrees C, and decreased at 36-39 degrees C. The pyknotic nuclei and fragmentation of the Sertoli cells were more obvious at > 36 degrees C. Western blot and immunofluorescence showed the highest level of OCLN expression at 36 degrees C, which, however, decreased while the temperature rose above 36 degrees C (P < 0. 01). CONCLUSION: High temperature (> 36 degrees C) inhibited the proliferation of rat Sertoli cells in vitro, and decreased the expression of OCLN, which suggests that a higher temperature above 36 degrees C may reduce male fertility by affecting the proliferation of Sertoli cells and integrity of the tight junction among Sertoli cells or Sertoli cells and other cells.

[Effects of Qiangjing Capsule on the oxidative and antioxidative system in the epididymis of varicocele rats].

OBJECTIVE: To observe the effects of Qiangjing Capsule (QC) on the oxidative amd antioxidative system in the epididymis of varicocele rats in comparison with those of Shaofuzhuyu Capsule (SC) and Wuziyanzong Capsule (WC), and to explore its possible mechanism of enhancing epididymal sperm maturation. METHODS: Ten of 100 adolescent male SD rats were randomized to a sham-operation group, and varicocele models were successfully established in 72 of the other 90 by narrowing of the left renal vein. Then the model rats were equally assigned to 6 groups: model control, high-dose QC (0.216 g/ml), medium-dose QC (0.108 g/ml), low-dose QC (0.054 g/ml), SC (0.146 g/ml), and WC (0.130 g/ml). After 4 weeks of treatment, we determined the activity of glutathione peroxidase (GPx) and the level of malondialdehyde (MDA) in the left epididymis of different groups of rats. RESULTS: Compared with the sham-operation group, the model group showed a significant decrease in GPx activity (P < 0.01) and a marked increase in the MDA level (P < 0.05). And the high-dose QC group exhibited a significantly hither GPx activity and lower MDA level than all the other groups (P < 0.01 and P < 0.05). CONCLUSION: Varicocele can reduce the activity of GPx and elevate the level of MDA in the epididymis of rats, while Qiangjing Capsule can increase the former and decrease the latter, and thereby may improve epididymal microenvironment, enhance epididymal sperm maturation and promote fertility.

Intervention with cilostazol attenuates renal inflammation in streptozotocin-induced diabetic rats.

AIMS: An inflammatory reaction is commonly found in the pathogenesis of diabetic nephropathy (DN). Cilostazol, a type 3 phosphodiesterase (PDE) inhibitor, has been previously reported to be anti-inflammatory, independent of an anti-platelet property. In the present study, we evaluated the hypothesis that cilostazol has protective effects on diabetic nephropathy by modulating the inflammatory process. MAIN METHODS: Cilostazol was administered (27 or 9 mg kg(-1)d(-1)) to streptozotocin (STZ)-induced diabetic rats for eight weeks. We studied the kidney expression of vascular cell adhesion molecule (VCAM)-1 and intercellular adhesion molecule (ICAM)-1 by immunofluorescence, western blotting and real-time PCR. The renal monocyte chemoattractant protein (MCP)-1 and vascular endothelial growth factor (VEGF) levels were examined by ELISA. The nuclear factor (NF)-kappaB-DNA binding activity was assessed by electrophoresis mobility shift assay (EMSA). KEY FINDINGS: Our results showed cilostazol inhibited diabetes-induced hypertrophy of the glomeruli and infiltration of inflammatory cells, as well as the increase in the VCAM-1 and ICAM-1 mRNA and protein expression, and MCP-1 and VEGF contents in the kidneys. Consistent with these findings, cilostazol attenuated the enhanced activation of NF-kappaB in diabetic rats. SIGNIFICANCE: These results demonstrate that the renoprotective effects of cilostazol may be mediated by its anti-inflammatory actions, including inhibition of NF-kappaB activation and the subsequent decrease in proinflammatory factors, such as VCAM-1, ICAM-1, MCP-1 and VEGF expression in kidneys of diabetic rats.

Tissue-specific expression of PPAR mRNAs in diabetic rats and divergent effects of cilostazol.

Cilostazol and ligands of peroxisome proliferator-activated receptors (PPARs) have been effectively used to alleviate diabetic complications, but the common and tissue-specific expression patterns of PPARs in different tissues in diabetic patients and those treated with cilostazol have not been reported. Here, we aimed to assess the effects of diabetes and cilostazol on mRNA expression of PPARalpha and PPARgamma in the aorta, renal cortex, and retina of diabetic rats treated with cilostazol for 8 weeks. PPARalpha mRNA expression showed uniform downregulation in all these tissues in diabetic rats, and this effect was reversed by cilostazol treatment. Surprisingly, PPARgamma mRNA expression was reduced in the renal cortex and retina, yet increased in the aorta of diabetic rats, although cilostazol still reversed these changes. Interestingly, cilostazol, a well-known phosphodiesterase 3 inhibitor and cAMP elevator, augmented cAMP content only in the aorta, but showed no significant effects in the renal cortex of diabetic rats. In conclusion, mRNA expression of PPARs is tissue-specific in diabetes and may be differently affected by cilostazol, possibly because of its tissue-specific effects on cAMP content.

Relationship between the expression of RASSF1A protein and promoter hypermethylation of RASSF1A gene in bladder tumor.

To investigate the relationship between the expression of RASSF1A protein and promoter hypermethylation of RASSF1A gene, RASSF1A protein expression was measured by Western blotting in 10 specimens of normal bladder tissues and 23 specimens of bladder transitional cell carcinoma (BTCC). The promoter methylation in BTCC and normal bladder tissues was detected by methylation-specific PCR (MSP). The results showed that the expression level of RASSF1A protein was significantly lower in BTCC tissues than that in normal bladder tissues. However, it was not correlated with its clinical stages and pathological grades. The frequency of promoter methylation of RASSF1A gene was higher in BTCC tissues than that in normal bladder tissues. In 14 patients with the aberrant promoter methylation, 13 showed loss or low expression of RASSF1A protein. It is concluded that RASSF1A gene promoter methylation may contribute to the low level or loss of RASSF1A protein expression, the inactivation of RASSF1A gene and the genesis of BTCC. But, it may bear no correlation with its clinical stages and pathological grades.

Relationship between the Expression of RASSF1A Protein and Promoter Hypermethylation of RASSF1A Gene in Bladder Tumor / 华中科技大学学报（医学）（英德文版）

To investigate the relationship between the expression of RASSFIA protein and promoter hypermethylation of RASSFIA gene, RASSFIA protein expression was measured by Western blot- ting in 10 specimens of normal bladder tissues and 23 specimens of bladder transitional cell carci- noma (BTCC). The promoter methylation in BTCC and normal bladder tissues was detected by me- thylation-specific PCR (MSP). The results showed that the expression level of RASSFIA protein was significantly lower in BTCC tissues than that in normal bladder tissues. However, it was not corre- lated with its clinical stages and pathological grades. The frequency of promoter methylation of RASSF1A gene was higher in BTCC tissues than that in normal bladder tissues. In 14 patients with the aberrant promoter methylation, 13 showed loss or low expression of RASSF1A protein. It is con- cluded that RASSFIA gene promoter methylation may contribute to the low level or loss of RASSFIA protein expression, the inactivation of RASSFIA gene and the genesis of BTCC. But, it may bear no correlation with its clinical stages and pathological grades.