Objective: To explore the association of zinc finger ZBTB22 gene single nucleotide polymorphisms (SNPs) and schizophrenia in Han Chinese population. Methods: A case-control study was designed by genotyping four SNPs (rs3130100, rs1061783, rs1061801 and rs3130099) in 658 schizophrenia patients and 658 healthy controls of Chinese Han origin. SHEsis was used to analyze genotypic and allelic distributions, linkage disequilibrium and haplotype distribution. Results: rs3130100, rs1061783, and rs3130099 showed significant differences between the cases and controls in allele frequencies (P<0. 05 after Bonferroni correction), but no statistically significant differences were found in genotype frequencies after Bonferroni correction. Strong linkage disequilibrium was found among the four SNPs, and the frequency of haplotype TCGA significantly decreased in the schizophrenia patients (P=0.015 after Bonferroni correction). Conclusion: The ZBTB22 SNPs rs3130100, rs1061783 and rs3130099 may be associated with schizophrenia in Han Chinese population.

Testicular prostheses have been used to deal with anorchia for nearly 80 years. Here, we evaluated a novel testicular prosthesis that can controllably release hormones to maintain physiological levels of testosterone in vivo for a long time. Silastic testicular prostheses with controlled release of testosterone (STPT) with different dosages of testosterone undecanoate (TU) were prepared and implanted into castrated Sprague-Dawley rats. TU oil was applied by oral administration to a separate group of castrated rats. Castrated untreated and sham-operated groups were used as controls. Serum samples from every group were collected to measure the levels of testosterone (T), follicle-stimulating hormone and luteinizing hormone (LH). Maximum intracavernous penile pressure (ICPmax) was recorded. The prostates and seminal vesicles were weighed and subjected to histology, and a terminal dexynucleotidyl transferase-mediated UTP nick end labeling (TUNEL) assay was used to evaluate apoptosis. Our results revealed that the weights of these tissues and the levels of T and LH showed significant statistical differences in the oral administration and TU replacement groups compared with the castrated group (P < 0.05). Compared with the sham-operated group, the ICPmax, histology and TUNEL staining for apoptosis, showed no significant differences in the hormone replacement groups implanted with medium and high doses of STPT. Our results suggested that this new STPT could release TU stably through its double semi-permeable membranes with excellent biocompatibility. The study provides a new approach for testosterone replacement therapy.

Objective: To establish a method for the isolation exosome from serum by polyethylene glycol (PEG). Methods: The ultracentrifugation, commercial EXOQuick precipitation kit and PEG8000 precipitation method were used to isolate exosomes from blood serum of the patients with breast cancer and benign breast lesions and the healthy individuals, respectively. The morphology and particle size of exosomes were measured by transmission electron microscopy and NanoSight NS90 method, respectively. The expression level of the surface marker CD9 in serum exosomes was determined by Western blotting. The expression level of microRNA-21 (miR-21) in serum samples and serum exosomes was detected by real-Time fluorescent quantitative PCR. Results: A method of isolating exosomes by PEG was established successfully. In terms of particle size distribution and CD9 concentration on the surface of exosomes, the performance of PEG8000 precipitation method was superior to ultracentrifugation, and equivalent to the commercial EXOQuick kit. Compared with breast benign lesions, the level of miR-21 was upregulated by 2.68 fold (P =0.024) in serum exosomes isolated from malignant breast cancer patients by using PEG8000 precipitation method. However, the miR-21 expression level was almost equal in serum samples from the patients with breast benign lesions and breast cancer (P =0.373). Conclusion: PEG8000 precipitation method for enriching exosomes is easy to operate, short of time and low in cost. The detection efficiency of miRNA expression in exosomes can be increased significantly than that in serum samples.

OBJECTIVE: To identify pathogenic mutation in a Chinese family affected with hereditary spastic paraplegia (HSP) through genetic testing and a follow-up survey. METHODS: Whole exome sequencing was performed on DNA samples of two patients and one unaffected member to screen candidate mutations. Sanger sequencing was used to validate the suspected mutations in all ten family members. RESULTS: Four patients and three asymptomatic members (under 25 years old) carried a c.1771T>C mutation of the KIAA0196, while the other three asymptomatic members (over 40 years old) did not carry the mutation. The mutation was predicted to be "affect protein function", "probably damaging" and "disease causing" by SIFT, PolyPhen-2 and Mutation Taster, respectively. Three asymptomatic carriers were followed up and one of them developed HSP one year later, while the other two had no signs of the disease yet. CONCLUSION The clinical phenotype of the c.1771T>C mutation of KIAA0196 has a considerable heterogeneity and this mutation may be a common pathogenic site of KIAA0196 mutations among Chinese patients with hereditary spastic paraplegia.
Adult ; Asian Continental Ancestry Group ; Heterozygote ; Humans ; Mutation ; Pedigree ; Phenotype ; Proteins ; genetics ; Spastic Paraplegia, Hereditary ; genetics

Histone variants confer chromatin unique properties and also participate in the regulation of genetic expression through specific deposition and removal machineries. macroH2A1 is one of histone variants and has been found implicating in female X chromosome inactivation and transcriptional repression. However, recent marcoH2A1 has been reported to suppress tumor proliferation through regulating cellular senescence and metabolism under specific situations. The paper reviews the roles of macroH2A1 in tumorigenesis and the molecular me-chanisms in regulating genetic expression based on current research progress.

Objective: To explore the relationship between the activity of Wnt/β-catenin signaling pathway and prostate cancer stem cells by utilizing a Wnt/β-catenin signaling reporter system. Methods: Immunofluorescence staining was used to explore the activation of Wnt/β-catenin signaling pathway in prostate cancer. Wnt/β-catenin signaling reporter plasmid 7TGP carrying T cell specific transcription factor (TCF) binding site and enhanced green fluorescent protein (GFP) was transfected into prostate cancer PC3 and DU145 cells by liposomes, respectively. Furthermore, the top 5% (GFP+) and bottom 5% (GFP-) groups of PC3 and DU145 cells according to the fluorescent intensity of GFP were collected via flow cytometry. Then Western blotting assay was used to compare the expression level of activated β-catenin protein in the nucleus of GFP+ and GFP- cell groups. The real-time fluorescent quantitative PCR was used to detect the expressions of Wnt/β-catenin signaling downstream target genes in GFP+ and GFPcell groups. Sphere formation assay and real-time fluorescent quantitative PCR were used to compare the stemness of GFP+ and GFP- cells. Results: The activity of canonical Wnt signaling pathway was low in most of prostate cancer cells, in which β-catenin was not activated and translocated to the nucleus. While β-catenin was increasingly translocated to the nucleus in a small percentage of prostate cancer cells resulting in high activation of Wnt signaling pathway. PC3-GFP+ and DU145-GFP+ cells had more expression of nuclear β-catenin comparing with the corresponding GFP- cells (both P < 0.05), and the expression level of Wnt signaling pathway down-stream target gene Axis inhibition protein 2 (AXIN2) was up-regulated (both P < 0.05). In addition, the expression levels of cancer stem cell markers including B-cell-specific moloney leukemia virusinsert site 1 (BMI1) and aldehyde dehydrogenase family 1 member A1 (ALDH1A1) were up-regulated in PC3-GFP+ and DU145-GFP+ cells comparing with the corresponding GFP- cells (all P < 0.05). The sphere formation capacity was remarkedly up-regulated in PC3-GFP+ and DU145-GFP+ cells comparing with the corresponding GFP- cells (both P < 0.05). Conclusion: Prostate cancer cells with higher activity of Wnt/β-catenin signaling show the enhanced stemness.

Objective: To investigate the effects of microRNA-378 (miR-378) on proliferation and migration of renal cancer cell lines in vitro, and to explore the possible mechanism. Methods: The expression of miR-378 in renal cell carcinoma and normal renal epithelial tissues was analyzed using the datasets from The Cancer Genome Atlas (TCGA). The expression of miR-378 in human renal carcinoma cell lines 786-O, ACHN, 769-P and Caki-1 as well as human renal proximal tubular epithelial cell line HK2 was detected by real-time fluorescent quantitative PCR (RFQ-PCR). MiR-378 mimic and inhibitor were synthesized and then transfected into renal carcinoma 786-O and Caki-1 cells, respectively. The proliferation and migration of 786-O and Caki-1 cells were detected by CCK-8 assay, Transwell chamber assay and wound healing assay, respectively. The potential target gene of miR-378 was screened by bioinformatics, and confirmed by RFQ-PCR and Western blotting in renal cancer cells, respectively. Results: The expression level of miR-378 in renal cancer tissues and cell lines was significantly up-regulated as compared with that in normal renal epithelial tissues or normal renal epithelial HK2 cells (all P < 0.001). After transfection with miR-378 mimic, the proliferation (P < 0.01) and migration (P < 0.001) of 786-O cells were remarkably increased. In contrast, after transfection with miR-378 inhibitors, the proliferation (P < 0.01) and migration (P < 0.001) of Caki-1 cells were remarkably suppressed. By bioinformatics assay, the large tumor suppressor 2 (LATS2) was identified as a target gene of miR-378 in renal cancer. The mRNA and protein expression of LATS2 were negatively related with the expression of miR-378 in renal cancer 786-O and Caki-1 cells (all P < 0.01). Conclusion: Overexpression of miR-378 may promote the proliferation and migration of renal cancer cells by negative targeting LATS2 gene.

Objective: To investigate the effects of Krüppel-like factor 4 (KLF 4) knockdown on epithelialmesenchymal transition (EMT), cell migration and invasion of prostate cancer cells. Methods: The prostate cancer cells with stable knockdown of KLF 4 named as LNCaP-shKLF4 cells and the LNCaP-con cells as the control were constructed. The expressions of KLF4 mRNA and protein in LNCaP-con and LNCaP-shKLF4 cells, and the expressions of EMTassociated gene mRNA and protein in the prostate cancer PC3-shKLF4 cells with stable knockdown of KLF 4 and the PC3-con cells (as the control) as well as the LNCaP-shKLF4 and LNCaP-con cells were detected by real-time fluorescence quantitative-PCR and Western blotting, respectively. The abilities of migration and invasion of PC3-shKLF4 and PC3-con cells as well as LNCaP-shKLF4 and LNCaP-con cells were detected by Transwell chamber assay. Results: LNCaP-shKLF4 and LNCaP-con cells were successfully constructed. The expression levels of KLF4 mRNA and proteins in LNCaP-shKLF4 cells were lower than those in LNCaPcon cells (P<0.01, P<0.05). The expression levels of E-cadherin (E-cad) mRNA in PC3- shKLF4 and LNCaP-shKLF4 cells were higher than those in PC3-con and LNCaP-con cells, respectively (both P<0.01). The expression levels of N-cadherin (N-cad), Zinc finger E-box-binding homeobox 1 (Zeb1), Snail1, vimentin (Vim) and matrix metallopeptidase 1 (MMP1) mRNAs in PC3-shKLF4 and LNCaP-shKLF4 cells were lower than those in PC3-con and LNCaP-con cells, respectively (all P<0.05). The expression levels of E-cad protein in PC3-shKLF4 and LNCaP-shKLF4 cells were higher than those in PC3-con and LNCaP-con cells, respectively (both P<0.05). The expression levels of N-cad, Zeb1, Snail1, Vim and MMP1 mRNAs in PC3-shKLF4 and LNCaP-shKLF4 cells were lower than those in PC3-con and LNCaP-con cells, respectively (all P<0.05). The abilities of migration and invasion of PC3-shKLF4 and LNCaP-shKLF4 cells were weaker than those of PC3-con and LNCaP-con cells, respectively (P<0.01, P<0.05). Conclusion: KLF 4 knockdown in prostate cancer cells can activate the expression of epithelium-associated genes and inhibit the expressions of mesenchymal-associated genes, resulting in the inhibition of cell migration and invasion of prostate cancer cells in vitro.

Although epigenetic regulation of histone modification has attracted more and more attention, the underlying mechanisms are still unclear. The present review introduces the role of acetylated H3 lysine 23 (H3K23ac) in glioma tumorigenesis, and demonstrates a new mechanism by which H3K23ac regulates gliomagenesis, which may provide novel approaches to glioma treatment.