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Background: Previous studies suggested that the H63D and C282Y polymorphisms in the HFE genes were susceptible to many cancer types, nevertheless, the present results were inconclusive. Thus, the present study was aimed to evaluate the association between the HFE polymorphisms (H63D and C282Y) and cancer risk via meta-analysis., Materials and Methods: We retrieved PubMed, Google Scholar, Embase and Web of Science databases for all eligible studies up to April 1, 2015. All the statistical analysis was conducted by STATA 12.0., Results: Finally, a total of 20 publications including 24 case-control studies, comprising 6,524 cases and 31,080 controls for HFE-C282Y polymorphism and 19 publications including 21 case control studies, comprising 5,648 cases and 14,257 controls for HFE-H63D polymorphism were enrolled in our analysis. An increased risk for overall cancer risk was identified in HFE-H63D polymorphism under allele contrast (D vs H: OR=1.153; 95%CI=1.031- 1.289, Pheterogeneity=0.002), homozygotes vs wide type (DD vs HH: OR=1.449; 95%CI=1.182-1.777, Pheterogeneity=0.391), dominant model (DD+HD vs HH: OR=1.145; 95%CI=1.007-1.301, Pheterogeneity=0.002) and recessive model (DD vs HD+HH: OR=1.416 ; 95%CI=1.156-1.735, Pheterogeneity=0.549), as well as HFE- C282Y under homozygotes vs wide type (YY vs CC: OR=1.428, 95%CI=1.017-2.006, Pheterogeneity=0.220). In addition, in the stratified analysis by cancer type, an increased risk was identified in hepatocellular carcinoma and breast cancer in C282Y polymorphism, as well as pancreatic cancer in H63D polymorphism, whereas a decreased risk of colorectal cancer was identified in C282Y polymorphism., Conclusions: Present study suggested that H63D and C282Y polymorphisms associated with an increased risk of overall cancer. Nevertheless, well- designed study with large sample size will be continued on this issue of interest.

Common genetic variation Q192R in the paraoxonase 1 (PON1) gene has been considered to be implicated in the development of many cancers. Nevertheless, results from the related studies were inconsistent. To elucidate the association, we performed a meta-analysis for 8,112 cases and 10,037 controls from 32 published case-control studies. Odds ratios (ORs) with 95% confidence intervals (CIs) were used to assess the strength of the association by STATA 12.0 software. Overall, we revealed that the PON1-192R allele was associated with a reduced risk of the overall cancers. Moreover, in the stratified analysis by cancer types (breast cancer, prostate cancer, brain cancer etc.), the results showed that PON1-192R allele was associated with a decreased risk in breast cancer (R vs Q: OR=0.605, 95% CI=0.378-0.967, Pheterogeneity=0.000; RR vs QQ: OR=0.494, 95% CI=0.275-0.888, Pheterogeneity=0.002; RQ vs QQ: OR=0.465, 95% CI=0.259-0.835, Pheterogeneity=0.000; and RR+RQ vs QQ: OR=0.485, 95% CI=0.274-0.857, Pheterogeneity=0.000), and associated with prostate cancer in homozygote (RR vs QQ: OR=0.475, 95% CI=0.251- 0.897, Pheterogeneity=0.001) and recessive models (RR vs RQ+QQ: OR=0.379, 95% CI=0.169-0.853, Pheterogeneity=0.000), while an increased risk was identified in lymphoma (R vs Q: OR=1.537, 95% CI=1.246-1.896, Pheterogeneity=0.944; RR vs QQ: OR=2.987, 95% CI=1.861-4.795, Pheterogeneity=0.350; RR+RQ vs QQ: OR=1.354, 95% CI=1.021-1.796, Pheterogeneity=0.824; and RR vs RQ+QQ: OR=2.934, 95% CI=1.869-4.605, Pheterogeneity=0.433), and an increased risk in prostate cancer under heterozygote comparison (RQ vs QQ: OR=1.782, 95% CI=1.077-2.950, Pheterogeneity=0.000) and dominant models (RR+RQ vs QQ: OR=1.281, 95% CI=1.044-1.573, Pheterogeneity=0.056). When subgroup analysis that performed by the control source (hospital based or population based), a decreased risk of the overall cancers was revealed by homozygote (RR vs QQ: OR=0.601, 95% CI=0.366-0.987, Pheterogeneity=0.000) and dominant models (RR vs RQ+QQ: OR=0.611, 95% CI=0.384-0.973, Pheterogeneity=0.000) in hospital based group. Stratifying by ethnicity, a significantly reduced risk of the overall cancers under allele contrast model (R vs Q: OR=0.788, 95% CI=0.626-0.993, Pheterogeneity=0.000) was uncovered in Caucasian. In summary, these findings suggested that PON1 Q192R polymorphism was associated with a reduced risk of the overall cancers, nevertheless, it might increase cancer susceptibility of prostate and lymphoma risk. Large well-designed epidemiological studies will be continued on this issue of interest.

Objective: To investigate the expression and significance of ubiquitously transcribed TPR gene on the X chromosome (UTX) in renal cell carcinoma (RCC) tissues, then to explore the relationship between UTX expression and renal cancer pathologic characteristics., Methods: In the study, 45 patients with dignosed renal cell carcinoma clinical samples were collected in Peking University Shenzhen Hospital. Total RNA and protein were extracted from the cancer tissues and adjacent normal tissues. UTX expression of cancer tissues and adjacent normal tissues was detected on both mRNA and protein levels using real time-PCR and IHC, respectively. And the relationship between UTX expression and the 45 patients' clinical characteristics was analyzed., Results: The mRNA level of UTX in cancer tissues(C) was 4.4 folds, higher than that of the adjacent normal tissues(N) [ 0.883 2±0.703 8 vs. 0.199 7±0.140 0, P<0.05]. The protein expression of UTX in cancer tissues was up-regulated, and the protein score of cancer tissues was 4 folds, change compared with adjacent normal tissues[12±4 vs. 3±3, P<0.05].The expression of UTX was associated with pathological grade(P=0.004)but without gender, age, tumor size and TNM stage., Conclusion: UTX is up-regulated in RCC tissues and the expression of UTX is associated with pathological grade, illustrating that UTX may play an important role in renal cancer progression.

Male infertility is a worldwide problem, and about 15% of the cases are associated with spermatogenesis-related gene mutation. The mammalian gene UBE2B is the homolog of the RAD6 gene of yeast, belonging to the ubiquitin proteasome system and playing an important role in spermatogenesis. Mice lacking the UBE2B gene are infertile, with reduced sperm motility, increased morphologically abnormal sperm, and inhibited meiosis of spermatogonia. Accumulated evidence shows that UBE2B gene mutants and single nucleotide polymorphisms are associated with male infertility. This article reviews the relation between the UBE2B gene and male infertility, offering some theoretical evidence for the diagnosis and treatment of male infertility.

The epithelial-to-mesenchymal transition (EMT), a process involving the breakdown of cell-cell junctions and loss of epithelial polarity, is closely related to cancer development and metastatic progression. While the cystic fibrosis transmembrane conductance regulator (CFTR), a Cl(-) and HCO3(-) conducting anion channel expressed in a wide variety of epithelial cells, has been implicated in the regulation of epithelial polarity, the exact role of CFTR in the pathogenesis of cancer and its possible involvement in EMT process have not been elucidated. Here we report that interfering with CFTR function either by its specific inhibitor or lentiviral miRNA-mediated knockdown mimics TGF-β1-induced EMT and enhances cell migration and invasion in MCF-7. Ectopic overexpression of CFTR in a highly metastatic MDA-231 breast cancer cell line downregulates EMT markers and suppresses cell invasion and migration in vitro, as well as metastasis in vivo. The EMT-suppressing effect of CFTR is found to be associated with its ability to inhibit NFκB targeting urokinase-type plasminogen activator (uPA), known to be involved in the regulation of EMT. More importantly, CFTR expression is found significantly downregulated in primary human breast cancer samples, and is closely associated with poor prognosis in different cohorts of breast cancer patients. Taken together, the present study has demonstrated a previously undefined role of CFTR as an EMT suppressor and its potential as a prognostic indicator in breast cancer., (© 2013.)

Optimization based on central composite design (CCD) for enantioseparation of anisodamine (AN), atenolol (AT), and metoprolol (ME) in human urine was developed using a microfluidic chip-CE device. Coupling the flexible and wide working range of microfluidic chip-CE device to CCD for chiral separation of AN, AT, and ME in human urine, a total of 15 experiments is needed for the optimization procedure as compared to 75 experiments using the normal one variable at a time optimization. The optimum conditions obtained are found to be more robust as shown by the curvature effects of the interaction factors. The developed microfluidic chip-CE-ECL system with adjustable dilution ratios has been validated by satisfactory recoveries (89.5-99% for six enanotiomers) in urine sample analysis. The working range (0.3-600 μM), repeatability (3.1-4.9% RSD for peak height and 4.0-5.2% RSD for peak area), and detection limit (0.3-0.6 μM) of the method developed are found to meet the requirements for bedside monitoring of AN, AT, and ME in patients under critical conditions. In summary, the hyphenation of CCD with the microfluidic chip-CE device is shown to offer a rapid means for optimizing the working conditions on simultaneous separation of three racemic drugs using the microfluidic chip-CE device developed., (© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

This study assesses the long-term outcomes in Han Chinese patients with clinical stage I non-seminomatous germ cell testicular cancer (CSI NSGCT) treated with surveillance, retroperitoneal lymph node dissection (RPLND) and adjuvant chemotherapy. We retrospectively evaluated 89 patients with a mean age of 26.5 years. After orchiectomy, 37 patients were treated with surveillance, 34 underwent RPLND and 18 were managed with chemotherapy. The overall survival rate, the recurrence-free survival rate and the risk factors were evaluated. The median follow-up length was 92 months (range: 6-149 months). Thirteen of the 89 patients (14.6%) had relapses, and one died by the evaluation date. The overall survival rate was 98.9%. The cumulative 4-year recurrence-free rates were 80.2%, 92.0% and 100% for the surveillance, RPLND and chemotherapy groups, respectively. The disease-free period tended to be briefer in patients with a history of cryptorchidism and those with stage Is. Therefore, surveillance, RPLND and adjuvant chemotherapy might be reliable strategies in compliant patients with CSI NSGCT. Surveillance should be recommended for patients with the lowest recurrence rate, especially those without lymphovascular invasion. This study might aid the establishment of a standard therapy for CSI NSGCT in China.

Renal plasmacytoma is extremely rare, presenting diagnostic challenges due to its unusual location and non-specific or absent symptoms. To the best of our knowledge, only 24 cases of renal plasmacytoma have been reported in the literature. The present study reports a case of primary renal plasmacytoma in a 46-year-old female patient. Computed tomography (CT) revealed that the mass was located in the lower pole of the left kidney and metastasis was detected in an enlarged para-aortic lymph node. Following careful preparation, a partial nephrectomy was performed and the retroperitoneal lymph node was resected. A pathological examination revealed a renal parenchyma with lymph node involvement; this was confirmed by immunohistochemistry and nested polymerase chain reaction (PCR). Consequently, a diagnosis of a renal extramedullary plasmacytoma (EMP) was proposed. Following this unexpected diagnosis, various examinations were performed, but there was no evidence of systemic plasma cell disease. The patient refused further therapy, including external beam radiotherapy and chemotherapy. Abdominal CT was performed three months post-surgery and did not reveal any relapse. The patient remains disease-free at nine months post-surgery. The current study also presents a review of the literature. Although the general prognosis and outcome of EMP is good, a follow-up examination is recommended due to the possibility of relapse or progression to plasma cell neoplasm (PCN).

Cystic fibrosis (CF) transmembrane conductance regulator (CFTR) is expressed in the epithelial cells of a wide range of organs/tissues from which most cancers are derived. Although accumulating reports have indicated the association of cancer incidence with genetic variations in CFTR gene, the exact role of CFTR in cancer development and the possible underlying mechanism have not been elucidated. Here, we report that CFTR expression is significantly decreased in both prostate cancer cell lines and human prostate cancer tissue samples. Overexpression of CFTR in prostate cancer cell lines suppresses tumor progression (cell growth, adhesion and migration), whereas knockdown of CFTR leads to enhanced malignancies both in vitro and in vivo. In addition, we demonstrate that CFTR knockdown-enhanced cell proliferation, cell invasion and migration are significantly reversed by antibodies against either urokinase plasminogen activator (uPA) or uPA receptor (uPAR), which are known to be involved in various malignant traits of cancer development. More interestingly, overexpression of CFTR suppresses uPA by upregulating the recently described tumor suppressor microRNA-193b (miR-193b), and overexpression of pre-miR-193b significantly reverses CFTR knockdown-enhanced malignant phenotype and abrogates elevated uPA activity in prostate cancer cell line. Finally, we show that CFTR gene transfer results in significant tumor repression in prostate cancer xenografts in vivo. Taken together, the present study has demonstrated a previously undefined tumor-suppressing role of CFTR and its involvement in regulation of miR-193b in prostate cancer development.

The mammalian SWI/SNF complex is one of ATP-dependent chromatin-remodeling complexes, which plays important roles in cell proliferation, differentiation, development and tumor suppression. ARID1A (AT-rich interactive domain-containing protein 1A) is a large subunit of SWI/SNF complex, and also an ARID family member with non- sequence-specific DNA binding activity. ARID1A is a tumor suppressor gene which is frequently mutated in many cancers, such as ovarian, bladder and gastric cancers. ARID1A can suppress cell proliferation through the up-regulation of p21 and the down-regulation of E2F-responsive genes. These findings on ARID1A and its role of tumor suppression contribute to understanding the mechanism of cancer development and developing new therapy for cancer.It is introduced in the review that ARID1A basic characteristic, related to cancer development, and biological role for full understanding of ARID1A.

Purpose: Serum cystatin C is a promising new biomarker of glomerular filtration rate and cardiovascular events, but few studies focused on serum cystatin C levels in obstructive sleep apnea (OSA) patients. The aim of our study was to evaluate the association between serum cystatin C and OSA in younger men (≤40 years old of age) without complications., Methods: We prospectively recruited consecutive participants without comorbidities who underwent polysomnography. Fasting blood samples were obtained from all subjects for biological profile measurements. Statistical analysis was used to evaluate the relationship between serum cystatin C and other parameters., Results: The population consisted of 98 subjects (mean age = 32.5 years, mean body mass index = 27.93 kg/m(2)) that were divided according to polysomnographic finding into control group (n = 23), mild (n = 15), moderate (n = 24), and severe (n = 36) OSA group. Compared with the control group, patients with severe OSA were significantly heavier (body mass index, 29.69 ± 3.81 vs. 26.42 ± 3.10) and presented significantly higher levels of high sensitive C-reactive protein (hsCRP) (1.10 ± 0.28 vs. 0.88 ± 0.20 mg/l) and serum cystatin C (0.87 ± 0.12 vs. 0.74 ± 0.10 mg/l) (p < 0.05 for all comparisons). Cystatin C was correlated with Apnea Hypopnea Index (AHI), Oxygen Desaturation Index, hsCRP, creatinine, and estimated glomerular filtration rate (r = 0.319, 0.279, 0.321, 0.233, -0.241, p = 0.001, 0.005, 0.001, 0.021, 0.017, respectively). After adjustment for confounding factors, AHI was significantly and positively associated with serum cystatin C levels (β = 0.284, p = 0.007)., Conclusions: Our finding indicated that serum cystatin C was associated with the severity of OSA in younger men. Further study is needed to find out whether OSA patients with increased serum cystatin C levels are prone to subclinical cardiovascular and renal diseases.

The aim of this study was to evaluate expression of COX-1 in renal cell carcinoma (RCC) and its prognostic value. mRNA of COX-1 was detected in 42 paired RCC and adjacent normal tissues with quantitative real- time polymerase chain reaction (qRT-PCR). Expression of COX-1 was also evaluated in 196 RCC sections and 91 adjacent normal tissues with immunohistochemistry. Statistical analysis was performed to assess COX-1 expression in RCC and its prognostic significance. The results of qRT-PCR showed mRNA levels of COX-1 in RCC tissues to be significantly higher than that in adjacent normal tissues (p < 0.001). Immunohistochemical assays also revealed COX-1 to be overexpressed in RCC tissues (p < 0.001). Statistical analysis demonstrated high expression of COX-1 was correlated with tumour size (p = 0.002), pathological stage (p = 0.003), TNM stage (p = 0.003, 0.007, 0.027, respectively), and tumour recurrence (p < 0.001). Survival analysis indicated patients with high expression of COX-1 had shorter survival time (p < 0.001), and COX-1 was an independent predictor. This is the first study to reveal overexpression of COX-1 in RRC and point to use as a prognostic marker in affected patients.

Azoospermia is one of the major reproductive disorders which cause male infertility in humans; however, the etiology of this disease is largely unknown. In the present study, six missense mutations of WT1 gene were detected in 529 human patients with non-obstructive azoospermia (NOA), indicating a strong association between WT1 mutation and NOA. The Wilms tumor gene, Wt1, is specifically expressed in Sertoli cells (SCs) which support spermatogenesis. To examine the functions of this gene in spermatogenesis, Wt1 was deleted in adult testis using Wt1(flox) and Cre-ER(TM) mice strains. We found that inactivation of Wt1 resulted in massive germ cell death and only SCs were present in most of the seminiferous tubules which was very similar to NOA in humans. In investigating the potential mechanism for this, histological studies revealed that the blood-testis barrier (BTB) was disrupted in Wt1 deficient testes. In vitro studies demonstrated that Wt1 was essential for cell polarity maintenance in SCs. Further studies found that the expression of cell polarity associated genes (Par6b and E-cadherin) and Wnt signaling genes (Wnt4, Wnt11) were downregulated in Wt1 deficient SCs, and that the expression of Par6b and E-cadherin was regulated by Wnt4. Our findings suggest that Wt1 is important in spermatogenesis by regulating the polarity of SCs via Wnt signaling pathway and that WT1 mutation is one of the genetic causes of NOA in humans., Competing Interests: The authors have declared that no competing interests exist.

The title compound, [Fe(C(12)H(8)N(2))(3)](C(6)H(5)N(2)O(2))(2), consists of one [Fe(phen)(3)](2+) cation (phen = 1,10-phenanthroline) and two 1,1-dicyano-2-eth-oxy-2-oxoethanide anions. Five atoms of the anion are disordered over two positions [site occupancy = 0.521 (13) for the major component]. In the complex cation, the Fe(II) atom is coordinated by six N atoms from three phen ligands in a distorted octa-hedral geometry. Two intra-molecular C-H⋯N hydrogen bonds occur in the complex cation. The crystal structure is mainly stabilized by Coulombic inter-actions. Weak intermolecular C-H⋯N inter-actions are also observed.

Chitosan, a cationic polysaccharide containing amino and hydroxyl groups, was used to fabricate an electrochemiluminescence (ECL) sensor. In the sensor construction, a glassy carbon electrode (GCE) was first coated by a chitosan film which embedded gold nanoparticles, and then the film was modified by introducing carboxyl groups on the surface, which were used to immobilize tris(2,2'-bipyridyl)ruthenium(II) doped amino-functional silica nanoparticles (NH(2)-RuSiNPs) through amido links. The successful modification was confirmed by scanning electronic microscopy and cyclic voltammetry. A binding model between the chitosan/nanogold composite film and NH(2)-RuSiNPs was also proposed, in which the amido link was the dominant bonding, accompanied with hydrogen bond interaction. ECL studies revealed that the sensor had very good response to different concentrations of 2-(dibutylamino) ethanol. This sensor was also applied in methamphetamine determination., (Copyright © 2012 Elsevier B.V. All rights reserved.)

atRA (all-trans-retinoic acid) is known to induce the differentiation of mESCs (mouse embryonic stem cells) into PGCs (primordial germ cells) in vitro. However, it is not clear as to what changes occur in PGC differentiation-associated genes or what mechanisms are involved when EBs (embryoid bodies) derived from mESCs are induced by atRA. EBs derived from mESCs were treated with 1, 2 or 5 μM atRA for 16 h, 2 days or 5 days. Real-time PCR and Western blot analysis were performed to detect the relative levels of PGC differentiation-associated genes (Lin28, Blimp1, Stra8 and Mvh) and the corresponding proteins respectively. Immunofluorescence was used to detect the protein location and distribution in EBs. The expression characteristics of genes could be divided into three categories: rapidly reached the peak value in 16 h and then decreased (Stra8, Lin28), initially low and then increased to reach the peak value in 5 days (Mvh) and relatively unchanged (Blimp1). A low level of Lin28 was expressed in EBs treated with atRA for 2 days or 5 days. The variation in the level of Lin28 mRNA did not influence the change in the level of Blimp1 mRNA. The changes in Stra8/Lin28 were consistent with the corresponding changes in the levels of their respective mRNAs, but the changes for Mvh/Blimp1 were not consistent with the corresponding changes in the levels of their respective mRNAs. Blimp1 expression may be independent of the effect of atRA on PGC differentiation. atRA may promote the start of a period in which there is a low level of Lin28 expression during PGC differentiation.

Androgen and androgen receptor (AR) play important roles in male spermatogenesis and fertility, yet detailed androgen/AR signals in Sertoli cells remain unclear. To identify AR target genes in Sertoli cells, we analyzed the gene expression profiles of testis between mice lacking AR in Sertoli cells (S-AR(-/y)) and their littermate wild-type (WT) mice. Digital gene expression analysis identified 2276 genes downregulated and 2865 genes upregulated in the S-AR(-/y) mice testis compared to WT ones. To further nail down the difference within Sertoli cells, we first constructed Sertoli cell line TM4 with stably transfected AR (named as TM4/AR) and found androgens failed to transactivate AR in Sertoli TM4 and TM4/AR cells. Interestingly, additional transient transfection of AR-cDNA resulted in significant androgen responsiveness with TM4/AR cells showing 10 times more androgen sensitivity than TM4 cells. In the condition where maximal androgen response was demonstrated, we then analyzed gene expression and found the expression levels of 2313 genes were changed more than twofold by transient transfection of AR-cDNA in the presence of testosterone. Among these genes, 603 androgen-/AR-regulated genes, including 164 upregulated and 439 downregulated, were found in both S-AR(-/y) mice testis and TM4/AR cells. Using informatics analysis, the gene ontology was applied to analyze these androgen-/AR-regulated genes to predict the potential roles of androgen/AR in the process of spermatogenesis. Together, using gene analysis in both S-AR(-/y) mice testis and TM4/AR cells may help us to better understand the androgen/AR signals in Sertoli cells and their influences in spermatogenesis.

The molecular mechanisms involved in the progression of clear cell renal cell carcinomas (ccRCCs) are still unclear. The aim of this study was to analyse the relationships between expression of RALYL and clinical characteristics. In 41 paired samples of ccRCCs and adjacent normal tissues, we used real-time qPCR to evaluate the expression of RALYL mRNA. RALYL protein levels were determined in 146 samples of ccRCC and 37 adjacent normal tissues by immunohistochemistry. Statistical analysis was used to explore the relationships between expression of RALYL and the clinical characteristics (gender, age, tumor size, T stage, N stage, M stage, survival times and survival outcome) in ccRCC. In addition, these patients were follow-up period 64 months (range: 4~116 months) to investigate the influence on prognosis. We found significantly differences between ccRCC tissues and normal tissues (p<0.001, paired-sample t test) in mRNA levels of RALYL. Immunohistochemistry analyses in 146 ccRCC samples and 37 adjacent normal tissues showed significantly lower RALYL protein levels in ccRCC samples (χ2-test, p<0.001), inversely correlating with tumour size (p=0.024), T stage (0.005), N stage (p<0.001) as well as M stage (p=0.019), but not age (p=0.357) and gender (p=0.348). Kaplan-Meier survival analysis demonstrated that people with lower level of RALYL expression had a poorer survival rate than those with a higher level of RALYL expression, significantly different by the log-rank test (p=0.011). Cox regression analysis indicated that RALYL expression (p=0.039), N stage (p=0.008) and distant metastasis (p<0.001) were independent prognosis factors for the overall survival of ccRCC patients. We demonstrated that the expression of RALYL was significantly low in ccRCC and correlated with a poor prognosis in a large number of clinical samples. Our findings showed that RALYL may be a potential therapeutic target as well as a poor prognostic factor.

Transcription factor 21 (TCF21) has been identified as a candidate tumor suppressor at 6q23-q24 that is epigenetically inactivated in many types of human cancers. We recently found that TCF21 methylation level was significantly increased in clear cell renal cell carcinoma (ccRCC). The purpose of this study was to investigate the prognostic impact of TCF21 expression in ccRCC and analyze the relationship between TCF21 expression and methylation level. We used real-time PCR and immunohistochemical staining to detect the expression of TCF21, and used methylation specific-PCR (MS-PCR) to determine the methylation status of TCF21 in ccRCC samples and cell line 786-O. The results showed that TCF21 expression level in ccRCC samples was significantly lower than in normal adjacent tissue samples (NAT samples). The Kaplan-Meier survival analysis demonstrated that TCF21 was a significant prognosticator of cancer-specific survival (p=0.001). Furthermore, the DNA demethylating agent 5'-azacytidine restored part of TCF21 expression by suppressing TCF21 methylation in 786-O. The methylation level of TCF21 in ccRCC samples was much higher than in NAT samples. These results suggest that the expression of TCF21 was an independent prognostic factor for poor survival in patients with ccRCC. Aberrant methylation was an important reason for the down-regulation the expression of TCF21, and may be associated with tumorigenesis in ccRCC.

Objective: To investigate whether mouse-induced pluripotent stem (iPS) cell line IP14D-1 has the potential to differentiate into induced primordial germ cells (iPGCs), and to explore the changes in the expression of iPGCs-differentiation associated genes and their possible mechanisms., Methods: Undifferentiated IP14D-1 was cultured to proliferate and then differentiated to form 4-, 7- and 9-day-old induced embryoid bodies (iEBs) in vitro, respectively. RT-PCR and immunofluorescence were used to detect the expressions of Lin28, Blimpl, Stra8 and Mvh, as well as the localization of the corresponding protein in iEBs., Results: The expression of Blimpl was higher than that of Lin28 in the undifferentiated IP14D-1 and mouse embryonic stem cells (mESCs). Mvh and Stra8 as well as mESCs and EBs were also expressed in IP14D-1 and iEBs, but with no significant differences. The expression of Lin28 was gradually increased in the IP14D-1-derived iEBs from 4 to 7 days, but decreased at 9 days, and the expression of Blimp1 was gradually reduced with the prolonged growing time of iEBs., Conclusion: A stable system was established for the culture and differentiation of IP14D-1 and IP14D-1-derived iEBs. The expressions of Lin28, Blimp1, Mvh and Stra8 were not significantly different between the undifferentiated IP14D-1 and mESCs, nor were the expressions of Mvh and Stra8 between iEBs and EBs. IP14D-1 and iEBs had the potential to differentiate into iPGCs, which increased in number in the 7-day-old iEBs, and the expression of iPGC-differentiation associated Lin28 became lower in the older iEBs.

Objective: To explore the role of microRNA-184(MIR-184) in the development of renal cell carcinoma(RCC)., Methods: The expressions of MIR-184 in 51 patients with RCC Investigated, normal adjacent tissues (ADTs) matched by fluorescence quantitative PCR technology (RT-qPCR) and the correlations analyzed between MIR-184 expression and the age, gender and clinical stage of RCC patients., Results: The average expression of MIR-184 in RCC was -14.664 6 ± 5.362 4, while that in ADTs was -10.408 7 ± 3.482 7(P<0.01). Bounded with the MIR-184 expression in RCC, patients were divided into lower-expression group and higher-expression group. Meanwhile, the RCC patients were divided into three groups according to the age, gender and clinical stage of the patients. Chi-square statistical analysis showed that the expression level of MIR-184 was not significantly correlated with the patient's age, gender and clinical stage (respectively: P>0.03, P>0.99, P>0.03)., Conclusion: MIR-184 in RCC was significantly lower than that in ADTs, which may have potential significance in the occurrence and development of RCC.

In this paper, an electrochemiluminescence (ECL) encoding method was developed for the first time, based on a dual-dye system including Ru(bpy)(3)(2+) as an ECL emitter and fluorescein isothiocyanate (FITC) as a coding dye.

The gene fusions between transmembrane protease serine 2 (TMPRSS2) and E26 (ETS) transcription factors are present in over 50% of patients with prostate cancer. TMPRSS2-ERG is the most common gene fusion type. The ERG overexpression induced by TMPRSS2-ERG gene fusion contributes to the development of prostate cancer. Both androgen receptor binding and genotoxic stress induce chromosomal proximity and TMPRSS2-ETS gene fusions. TMPRSS2-ERG gene fusion functions as a biomarker for prostate cancer, which can be easily detected in urine. This review focuses on the characteristics, oncogenic and rearranged mechanism, and clinical application of TMPRSS2-ETS gene fusions.

We performed a genome-wide association study of esophageal squamous cell carcinoma (ESCC) by genotyping 1,077 individuals with ESCC and 1,733 control subjects of Chinese Han descent. We selected 18 promising SNPs for replication in an additional 7,673 cases of ESCC and 11,013 control subjects of Chinese Han descent and 303 cases of ESCC and 537 control subjects of Chinese Uygur-Kazakh descent. We identified two previously unknown susceptibility loci for ESCC: PLCE1 at 10q23 (P(Han combined for ESCC) = 7.46 x 10(-56), odds ratio (OR) = 1.43; P(Uygur-Kazakh for ESCC) = 5.70 x 10(-4), OR = 1.53) and C20orf54 at 20p13 (P(Han combined for ESCC) = 1.21 x 10(-11), OR = 0.86; P(Uygur-Kazakh for ESCC) = 7.88 x 10(-3), OR = 0.66). We also confirmed association in 2,766 cases of gastric cardia adenocarcinoma cases and the same 11,013 control subjects (PLCE1, P(Han for GCA) = 1.74 x 10(-39), OR = 1.55 and C20orf54, P(Han for GCA) = 3.02 x 10(-3), OR = 0.91). PLCE1 and C20orf54 have important biological implications for both ESCC and GCA. PLCE1 might regulate cell growth, differentiation, apoptosis and angiogenesis. C20orf54 is responsible for transporting riboflavin, and deficiency of riboflavin has been documented as a risk factor for ESCC and GCA.

The spermatogenesis failure with a genetic defect is one of the major causes of male infertility. The Y chromosome is considered a lack of important functional genes. It was the discovery of the sex determining region Y that rekindled scientists'attention to the Y chromosome. The successful sequencing of the Y chromosome uncovered its actual structure and the molecular base of its microdeletion. Of the 220 Y chromosome genes (104 coding genes, 111 pseudogenes, and 5 other uncategorized genes), 16 coding genes have been found in the azoospermia factor region (AZF) and related with male fertility. To date, more than 12 Y chromosome microdeletions have been discovered in the AZF region. The amplicons regions in the Y chromosome are the genetic base of microdeletion occurrence. The Y chromosome microdeletions in the AZF region have been identified as a relatively common cause of male infertility and diagnosed by multiplex PCR in the clinical laboratory. Genomics has brought many revolutionized tools beneficial for better understanding the genetics of mal infertility and defining the role of the Y chromosome gene in spermatogenesis.

A nonenzymatic electrochemical method was developed for glucose detection using an electrode modified with palladium nanoparticles (PdNPs)-functional carbon nanotubes (FCNTs). PdNPs were homogeneously modified on FCNTs through a facile spontaneous redox reaction and characterized by transmission electron microscopy. Based on the voltammetric and amperometric results, PdNPs efficiently catalyzed the oxidation of glucose at 0.40 V in the presence of 0.2M NaCl and showed excellent resistance towards poisoning from such interfering species as ascorbic acid, uric acid, and p-acetamidophenol. This anti-poisoning ability was investigated using analysis of the electrocatalytic products by in situ subtractively normalized interfacial Fourier transform infrared reflection spectroscopy, and the results indicated that no strongly adsorbed CO(ad) species could be found in the oxidation products, which was obviously different from the results obtained using Pt-based electrodes. In order to verify the sensor reliability, it was applied to the determination of glucose in urine samples. The results indicated that the proposed approach provided a highly sensitive, wide linear range, more facile method with good reproducibility for glucose determination, promising the development of Pd-based material in nonenzymatic glucose sensing., ((c) 2010 Elsevier B.V. All rights reserved.)

Small RNAs can silence transposons at transcriptional or post-transcriptional level. Newly identified small RNAs (e.g., piRNAs and endo-siRNAs) can repress the activity of transposons. Drosophila piRNAs, mouse piRNAs, mouse endo-siRNAs, and their related proteins are expressed primarily within the germline. This suggests that the small RNA pathway plays an important role in transposon silencing of the germline. Recent studies revealed the connection between small RNAs, transposon silencing, and the regulation of fertility, but the mechanism has not been clarified in detail. In this review, the advances in the control of genomic transposon activity and the regulation of fertility by the small RNA pathway were summarized and discussed.

We performed a genome-wide association study (GWAS) of systemic lupus erythematosus (SLE) in a Chinese Han population by genotyping 1,047 cases and 1,205 controls using Illumina Human610-Quad BeadChips and replicating 78 SNPs in two additional cohorts (3,152 cases and 7,050 controls). We identified nine new susceptibility loci (ETS1, IKZF1, RASGRP3, SLC15A4, TNIP1, 7q11.23, 10q11.22, 11q23.3 and 16p11.2; 1.77 x 10(-25) < or = P(combined) < or = 2.77 x 10(-8)) and confirmed seven previously reported loci (BLK, IRF5, STAT4, TNFAIP3, TNFSF4, 6q21 and 22q11.21; 5.17 x 10(-42) < or = P(combined) < or = 5.18 x 10(-12)). Comparison with previous GWAS findings highlighted the genetic heterogeneity of SLE susceptibility between Chinese Han and European populations. This study not only advances our understanding of the genetic basis of SLE but also highlights the value of performing GWAS in diverse ancestral populations.

Objective: Interactions of cells with the extracellular matrix (ECM) are essential for cell differentiation. The authors sought to determine the roles of different ECMs in the expressions of germ cell differentiation associated genes after mouse embryonic stem cells (mESCs) differentiated into embryoid bodies (EBs)., Methods: EBs derived from mESCs were maintained in suspension for 3 days and then cultured on the plates coated with various ECMs, including fibronectin (F), laminin (L), matrigel (M), collagen (C) and nonadhensive agarose (A), respectively, for 1, 2, 3 or 4 days, followed by evaluation of the expressions of the genes associated with germ cell differentiation by RT-PCR., Results: The EBs of the F and L groups exhibited facilitated adherent differentiation. The expressions of the Blimp-1, Stella, Mvh and Stra8 genes were increased gradually in the F and L but not obviously in the M and C groups. The overall gene expressions were low in the A group, but high and then gradually decreased in the blank control group. Endogenous fibronectin, laminin and integrin beta1 were obviously expressed in the L and control groups., Conclusion: Laminin /integrin beta1 signaling may play a role in regulating the differentiation of mESCs into primordial germ cells (PGCs). Exogenous laminin can facilitate the differentiation of mESC-derived EBs into PGCs by acting on the integrin beta1 subunit, while exogenous fibronectin may be involved in the regulation of the differentiation through other integrin subunit. Endogenous laminin and fibronectin secreted by EBs may also facilitate cell differentiation in the absence of exogenous ECMs.

Objective: To compare the expressions of ODF1 (outer dense fiber of the sperm tail 1) in ejaculated spermatozoa from normozoospermic and asthenozoospermic men with low sperm motility., Methods: Semen analyses were performed on the semen samples obtained from normozoospermic (n=20) and asthenozoospermic (n=20) volunteers according to the WHO criteria. To rule out the contamination of germ cells and leucocytes, the human ejaculated spermatozoa were purified by a discontinuous Percoll density gradient centrifugation. RT-PCR and Western blot were used to detect the expressions of ODF1 in the spermatozoa from the two groups., Results: RT-PCR showed that the expression of ODF1 mRNA was significantly lower in the spermatozoa from the asthenozoospermic patients than in those from the normozoospermic men (1.35 +/- 0.25 vs. 2.79 +/- 0.28, P < 0.05). Western blot confirmed the results from RT-PCR and revealed an obviously decreased expression of ODF1 in the spermatozoa of the asthenozoospermic patients, with statistically significant difference from the normozoospermic group (1.44 +/- 0.26 vs. 3.64 +/- 0.34, P < 0.05)., Conclusion: The expression of ODF1 was significantly decreased in the ejaculated spermatozoa of asthenozoospermic men, which might be responsible for low sperm motility.

An ethanol biosensor, based on the electrogenerated chemiluminescence of Ru(bpy)(3)(2+)-doped silica nanoparticles (RuSiNPs), was investigated in this study. The biosensor was a modified glassy carbon electrode, where alcohol dehydrogenase was crosslinked to RuSiNPs, and then immobilized on the electrode surface using chitosan. The results indicated that the biosensor exhibited excellent performance during ethanol determination with a wide linear range (10(-7) to 10(-2) M), low detection limit (5.0x10(-8) M) and good stability.

In this study, a novel material, palladium nanoparticles-carboxylic functional carbon nanotubes (PdNPs-CFCNTs), based on PdNPs supported on CFCNTs was synthesized by a facile spontaneous redox method. The material reveals high electrochemical activity and excellent catalytic characteristic for alcohol electrooxidation on a glassy carbon electrode (GCE) in an alkaline medium. The preparation mechanism was studied by the galvanic cell effect between PdCl(4)(2-) and functional defect sites on CFCNTs. Results from UV-visible absorption spectroscopy and electrochemical impedance spectroscopy revealed that the reduction of PdCl(4)(2-) to metallic Pd was successfully achieved. Morphologies of PdNPs supporting on CFCNTs (PdNPs-CFCNTs) were also characterized by transmission electron micrograph. PdNPs-CFCNTs with the best electrocatalytic characteristics were obtained under the condition as: the weight ratio of Pd to CFCNTs was kept at 2:1, the temperature was kept at 70 degrees C in the synthesis, and the scan rate of the applied potential was selected at 60 mV s(-1). The results indicate that PdNPs-CFCNTs could be a great potential material in direct ethanol fuel cells and ethanol sensors.

Ropporin has been identified as a spermatogenic cell-specific protein and may be involved in sperm maturation, motility, capacitation, hyperactivation and acrosome reaction. However, latest studies have shown that Ropporin is expressed weakly in normal non-testis tissues and highly in hematologic malignancies. Its highly conservative expression in mammalians demonstrates its importance to life. This paper updates the characterization, expression and its distribution, and biological function of Ropporin, and the advances in the clinical researches of the protein.

A novel non-enzymatic electrochemiluminescence (ECL) sensor based on palladium nanoparticles (PdNPs)-functional carbon nanotubes (FCNTs) was discovered for glucose detection. PdNPs were homogeneously modified on FCNTs using a facile spontaneous redox reaction method. Their morphologies were characterized by transmission electron microscopy (TEM). Based on ECL experimental results, the PdNPs-FCNTs-Nafion film modified electrode displayed high electrocatalytic activity towards the oxidation of glucose. The free radicals generated by the glucose oxidation reacted with the luminol anion (LH(-)), and enhanced the ECL signal. Under the optimized conditions, the linear response of ECL intensity to glucose concentration was valid in the range from 0.5 to 40 micromol L(-1) (r(2)=0.9974) with a detection limit (S/N=3) of 0.09 micromol L(-1). In addition, the modified electrode presented high resistance towards the poisoning of chloride ion, high selectivity and long-term stability. In order to verify the sensor reliability, it was applied to the determination of glucose in glucose injection samples. The results indicated that the proposed approach provided a highly sensitive, more facile method with good reproducibility for glucose determination, promising the development of a non-enzymatic ECL glucose sensor.

The formation of embryoid bodies (EBs) is the principal step in the differentiation of embryonic stem (ES) cells. In this study, the morphological characteristics and gene expression patterns of EBs related to the sequential stages of embryonic development were well defined in four distinct developmental groups over 112 days of culture: early-stage EBs groups (1-7 days of differentiation), mid-stage EBs groups (9-15 days of differentiation), maturing EBs groups (17-45 days of differentiation) and matured EBs groups (50 days of differentiation). We first determined definite histological location of apoptosis within EBs and the sequential expression of molecular markers representing stem cells (Oct4, SSEA-1, Sox-2 and AKP), germ cells (Fragilis, Dazl, c-kit, StellaR, Mvh and Stra8), ectoderm (Neurod, Nestin and Neurofilament), mesoderm (Gata-1, Flk-1 and Hbb) and endoderm (AFP and Transthyretin). Our results revealed that developing EBs possess either pluripotent stem cell or germ cell states and that three-dimensional aggregates of EBs initiate mES cell differentiation during prolonged culture in vitro. Therefore, we suggest that this EB system to some extent recapitulates the early developmental processes occurring in vivo.

Objective: To identify the genes involved in sperm motility., Methods: We hybridized asthenospermia and normal motile sperm cDNA samples with the human whole genome Affymetrix chip to screen differentially expressed genes. Then we detected the mRNA expressions of the voltage-dependent anion channel genes (VDACs) in human organs and spermatozoa by RT-PCR and compared their expressions in the poor and normal motility spermatozoa., Results: Differentially expressed genes VDACs were identified by analysis of the hybridization signals, including the 3 subtypes VDAC1, VDAC2 and VDAC3. The expression of VDAC2 mRNA was significantly decreased in the poor motility sperm (0.568 +/- 0.036), as compared with the healthy men (0.803 +/- 0.043, P < 0.01)., Conclusion: The decreased expression of VDAC2 in the ejaculated spermatozoa is possibly associated with the reduction of sperm motility.

Microdeletion of the azoospermia factor in the Yq of the Y chromosome is one of the important causes of male infertility. Complete deletion of the AZFc region (b2/b4 deletion) is the most common type of AZF deletion. Recent studies have shown a variety of deletions of the AZFc region, including partial deletions, such as gr/gr deletion, b1/b3 deletion and b2/b3 deletion, which may also be associated with male infertility.

Previous studies showed that gossypol could block the gap junctional intercellular communication (GJIC) between cultured cells. The present study was designed to investigate the effects of gossypol on the GJIC and the expression of connexin43 (Cx43) in the cultured cells. A Sertoli cell line, TM4, was treated with different concentrations of gossypol 1.25, 2.5, 5, and 10micromol/L for 6, 12, 24, and 48h. Cell viability was assessed with CCK-8 assay. GJIC in the cells was determined using the scrape loading and dye transfer (SLDT) assay; the expression of Cx43 was detected by RT-PCR, immunofluorescence and Western blot analysis. The SLDT assay showed gossypol significantly decreased GJIC between adjacent cells. RT-PCR, immunofluorescence and Western blot analyses demonstrated the expression of Cx43 in TM4 cells. The expression of Cx43 was gradually decreased with the increasing concentrations of gossypol, and the effect occurred as early as 6h after the treatment and continued until 48h. These results suggested that gossypol impaired GJIC by decrease of Cx43 expression in the cells, which is important for Sertoli cells to regulate spermatogenesis.

A simple, novel electrochemiluminescence (ECL) method for the detection of dichlorvos pesticide (DDVP) with high sensitivity was discovered. Detection was carried out in a static ECL system, in which a glassy carbon electrode was selected as the working electrode. ECL parameters, including the concentrations of cetyrltrimethylammonium bromide and luminol, the solution pH, and the scan rate of the applied potential, were optimized. Under these optimal conditions, the linear response of ECL-emission versus DDVP concentration was valid in the range 5-8000 ng/L (r(2)=0.9982) with a relative standard deviation of 4.3% at 2000 ng/L (n=10), yielding a detection limit (S/N=3) of 0.42 ng/L. The ECL emission was caused by a radical reaction process, in which the dissolved oxygen in the luminol solution reacted with the DDVP and generated free radicals. The free radicals reacted with the luminol anion and yielded the luminol radical. The approach presented was successfully applied to the determination of DDVP residues in vegetable samples.

Objective: To investigate the expression of amphiregulin in human endometrium during the menstrual cycle., Methods: Endometrial tissues were collected from the patients undergoing hysterectomy or endometrial biopsy. Real-time RT-PCR, in situ hybridization and immunohistochemistry were used to detect the expression characteristics of amphiregulin in human endometrium in proliferative and secretory phases., Results: Real-time RT-PCR showed the expression of amphiregulin mRNA in secretory phase was 32 times that in proliferative phase. The results from in situ hybridization and immunohistochemistry showed that amphiregulin was located in the cytoplasm and mainly expressed in the gland of endometrium. The expressions of amphiregulin mRNA in proliferative and secretory phases were 0.54+/-0.22 and 2.96+/-0.47 (P<0.01), and the expressions of amphiregulin protein in proliferative and secretory phases were 0.77+/-0.47 and 2.60+/-0.43 (P<0.01), respectively. The data demonstrated that the amphiregulin expression was significantly increased in secretory phase, which was consistent with the results from RT-PCR., Conclusion: The expression of amphiregulin was increased in the secretory phase of human endometrium, which suggests that amphiregulin may play an important role in human implantation.

Aim: To detect the expression of VASA in human ejaculated spermatozoa, and to compare the expression of VASA between normozoospermic men and patients with oligozoospermia., Methods: Ejaculated spermatozoa were collected from normozoospermic men and patients with oligozoospermia by masturbation, and subsequently segregated through a discontinuous gradient of Percoll to obtain the spermatozoa. Reverse transcription polymerase chain reaction (RT-PCR), quantitative RT-PCR (QRT-PCR), immunoflurescence and Western blotting were used to detect the expression of VASA in mRNA and protein levels., Results: VASA mRNA was expressed in the ejaculated spermatozoa. QRT-PCR analysis showed that VASA mRNA level was approximately 5-fold higher in normozoospermic men than that in oligozoospermic men. Immunofluorescence and Western blotting analysis showed that VASA protein was located on the cytoplasmic membrane of heads and tails of spermatozoa, and its expression was significantly decreased in oligozoospermic men, which is similar to the result of QRT-PCR., Conclusion: The expression of VASA mRNA and protein was significantly decreased in the sperm of oligozoospermic men, which suggested the lower expression of the VASA gene might be associated with pathogenesis in some subtypes of male infertility and VASA could be used as a molecular marker for the diagnosis of male infertility.

Previous studies showed that interleukin-6 (IL-6) was expressed in human Leydig and Sertoli cells and that it inhibited sperm motility. The aim of this study was to compare the expression of IL-6, IL-6R, and GP130 in ejaculated spermatozoa between normozoospermic and asthenozoospermic men. Human spermatozoa in the semen were purified by Percoll gradient technique to separate the seminal plasma and other round cells. RT-PCR, immunocytochemistry, and Western blot were used to detect the expression of IL-6, IL-6R, and GP130 in spermatozoa. With RT-PCR, only GP130 mRNA but not IL-6 and IL-6R mRNA was expressed in human ejaculated spermatozoa. The expression of GP130 mRNA was significantly lower in asthenozoospermic men than in normozoospermic men. The protein expression of GP130 was further confirmed by both immunocytochemistry and Western blot. Again, GP130 protein levels were significantly lower in asthenozoospermic men than in normozoospermic men. The results suggested that the decreased expression of GP130 in ejaculated spermatozoa could be associated with low sperm motility in asthenozoospermic men.

Since Nature published the first report in 2002 on using immunodeficient mice as recipients and allogeneous or heterogeneous testes as donor tissues to study the ectopic development of spermatogenic cells, the technique has been widely applied in various species (including human). In comparison with other in vitro maturation methods for male germ cells, testicular allografting or xenografting technique has such advantages as similar environment for the development of germ cells in physiological conditions, and better reproducibility. Up to now, sperm has been successfully produced by this technique from the testicular tisues of the immature mouse, hamster, cat, rabbit, pig, goat, bovine and rhesus monkey, and their offspring have even been generated by ICSI technique using the mouse and rabbit sperm derived from testis grafts. This article comprehensively reviews the development of the technique by discussing the influencing factors on the germ cell development in grafts including the variety and age of donors, the sex, integrity and immunity of recipients, the graft location and grafting time. And the applications of the technique and the existing problems are discussed as well.

Objective: To establish the method to identify human testicular spermatogenic cells., Methods: Cells were dispersed by mechanic disintegration on testicular biopsy samples of obstructive azoospermic patients. Diff-Quik staining was applied to mixed cell smears. Live cells were observed under inverted microscope equipped with Hoffman modulation contrast optics, and classified according to their morphology. Chromogenic in situ hybridization (CISH) using chromosome 17 centromere probe and anti-c-kit immunocytochemistry staining were applied to classified cell smears., Results: Sertoli cell, primary pachytene spermatocyte (PPS), spermatogonia, round spermatid were the main four round cell groups that can be classified under Hoffman optics. In CISH, Sertoli cell, PPS and spermatogonia displayed 2 centromere signals, while round spermatid and elongated spermatid/sperm displayed 1 centromere signal. In immunocytochemistry staining, PPS and spermatogonia displayed positive staining, while Sertoli cell, round spermatid and sperm displayed negative staining., Conclusion: Dispersed human testicular cells displayed different characteristics in live/staining morphology, ploidy analysis, cell surface membrane antigen expression. All of these methods can be chosen to identify human testicular spermatogenic cells at different stages, while cell morphology classifying under Hoffman optics is a simple and effective method for live cell identification.

Objective: To investigate the protein expression of hypoxia inducible factor-1alpha (HIF-1alpha) and glucose transporter protein 1 (Glut-1) in renal cancer and bladder cancers and the clinical significance thereof., Methods: TransAMTMELISA was used to detect the protein expression of HIF-1alpha in the tissue sections from 25 patients with bladder transitional cell carcinoma (TCC) and 16 patients with renal clear cell carcinoma resected during operation and small amounts of normal renal and bladder tissues far from the cancer tissue resected simultaneously. Immunohistochemistry was used to detect the protein expression of Glut-1 in tissue sections from 58 patients with bladder carcinoma transitional cell carcinoma and 38 patients with renal clear cell carcinoma resected during operation. 16 specimens of normal bladder and 16 specimens of normal renal tissue were obtained from the patients with other diseases who underwent operation during the same period to be used as controls. Follow-up was conducted for more than 2 years for all patients., Results: The protein expression of HIF-1alpha in the tissue of renal clear cell carcinoma was 3.38 +/- 1.71 microg/well, significantly higher than that in the tissue near the cancer (2.23 +/- 1.07 microg/well, P < 0.01). The protein expression of HIF-1alpha in the bladder TCC tissue was 2.69 +/- 1.34 microg/well, not significantly different from that in the tissue near the cancer (248 +/- 1.28 microg/well, P = 0.60). Protein expression of Glut-1 was not detected in the normal renal and bladder tissues, however, was significantly higher in the bladder TCC and renal clear cell carcinoma tissues. The positive rate of Glut-1 in the bladder TCC tissue was 77.90% (45/58). The positive rates of Glut-1 in the tissues of bladder TCC tissue, grades G1, G2, and G3 were 66.7%, 89.1%, and 53.3%. (P = 0.29), showing the correlation of Glut-1 expression with the grading of cancer. The positive rate of Glut-1 in the superficial TCC was 83.9%, not significantly different from that of the invasive TVV (70.4%, P = 0.90), showing that the Glut-1 is not correlated with the cancer staging. Nineteen of the 31 cases of superficial TCC showed recurrence within 2 years, however, the protein expression of Glut-1 rate was not significantly different between those with recurrence and those without recurrence (P = 0.90), showing that the protein expression of Glut-1 is not correlated with the recurrence of TCC. The protein expression of Glut-1 in the renal clear cell carcinoma was at a rate of 86.9% (33/38), however, it was not correlated with the grade and stage of the cancer., Conclusion: The protein expression of HIF-1alpha is strongly associated with the neoplastic progression of renal clear cell carcinoma, but its role in the development of bladder TCC is not clear yet. The protein expression of Glut-1 is strongly associated with the neoplastic progression of bladder TCC and renal clear cell carcinoma.

Aim: To investigate the stepwise development and germ cell gene expression in allografted neonatal mouse testes and the differentiation of immature human testicular cells in xenografted human testes., Methods: Immunodeficient nude mice were used as hosts for allografting of neonatal mouse testes and xenografting of human fetal testicular tissues. Stepwise development and stage-specific gene expression of germ cells in allografts were systematically evaluated and parallel compared with those in intact mice by periodically monitoring the graft status with measurement of graft weight, histological analysis and determination of five stage-specific genes. Human testicular tissues from 20 and 26 weeks fetuses were used for the xenografting study. Histological analysis of xenografts was performed 116 and 135 d after the grafting procedure., Results: In the allografting study, progressive increase in tissue volume and weight as well as in tubule diameter in grafts was observed; the appearance time of various germ cells in seminiferous tubules, including spermatogonia, spermatocytes, round and elongate spermatids and sperm, was comparable with that in intact donors; the initiation of gene transcription in grafts showed a similar trend as in normal mice. Graft weight ceased to increase after 7-8 weeks and degradation of grafts was observed after 5 weeks with progressive damage to seminiferous epithelium. In the xenografting study using immature human testicular tissues, graft survival and development was indicated by increasing graft weight, Sertoli cells differentiation into advanced stage, germ cells migration and location to the basal lamina and formation of a niche-like structure., Conclusion: The developmental course and gene expression pattern of germ cells in allografts were similar to those in intact mice. The best time point for retrieval of mouse sperm from grafts was 5-7 weeks after grafting procedure. An accelerated development of immature human testicular cells could be achieved by ectopic xenografting of human testes.

Objective: To investigate the differentiation of human testicular spermatogenic cells during in vitro culture., Methods: Testicular cells of obstructive azoospermic patients' testis biopsies were dispersed employing mechanic methods. Then, (1) mixed testicular cells were applied to in vitro culture, and changes of the ratio of elongating spermatids and all round cells were analyzed during mixed cell culture; (2) round spermatids were picked up from the mixed cells employing micromanipulator, followed by differentiation of the isolated round spermatids during microdrop culture., Results: The ratio of the elongating spermatids increased significantly (P < 0.05) after 24 hours of mixed cell culture in HTF medium supplemented with FSH and testosterone. During single round spermatid culture, transformation of the round spermatid to elongating spermatid with newly formed flagellum was observed, and the transformation ratio within 48 hours of microdrop culture was 3.54%. The differentiation of human testicular spermatogenic cells cultured in Vero cell conditioned medium was similar to that cultured in HTF medium., Conclusion: Human testicular round spermatids can differentiate to elongating spermatids during in vitro culture. Vero cell conditioned medium does not promote the differentiation of human testicular round spermatids to elongating spermatids.

Objective: To evaluate the predictive value of serum inhibin B (INH B) levels as an indicator of the presence of testicular spermatozoa in nonobstructive azoospermia., Methods: Forty patients with nonobstructive azoospermia (NOA), 20 patients with obstructive azoospermia (OA), and 10 fertile volunteers were involved in this study. A chemoluminescence method was used to measure the levels of FSH; Inhibin B was analysed by using sandwich enzyme-linked immuno-sorbent assay., Results: Patients with nonobstructive azoospermia has significantly higher levels of serum FSH [(21.34 +/- 12.15) IU/L] and significantly lower levels of inhibin B [(53.15 +/- 58.74) ng/L] than patients with obstructive azoospermia [FSH: (3.94 +/- 1.52) IU/L, INH B: (162.49 +/- 78.38) ng/L, P < 0.01] and fertile volunteers [FSH: (4.27 +/- 2.84) IU/L, INH B: (228.49 +/- 110.68) ng/L, P < 0.01]. Mean serum inhibin B were significantly higher in patients with nonobstructive azoospermia who had spermatozoa on TESE than in those in whom no spermatozoa was found on TESE [INHB: (90.31 +/- 72.18) ng/L vs (19.54 +/- 20.38) ng/L, r = 0.528, P < 0.01], but mean FSH levels did not have similar predictive power (P > 0.05)., Conclusion: Serum INH B level seems to be more accurate than serum FSH in the prediction of presence of testicular spermatozoa in patients with nonobstructive azoospermia. Serum inhibin B determination may be substitute of TESE as a diagnostic index.