LncRNA HOTAIR regulates the expression of E-cadherin to affect nasopharyngeal carcinoma progression by recruiting histone methylase EZH2 to mediate H3K27 trimethylation.

BACKGROUND/AIM: There has been increasing evidence for the function of long non-coding RNA (lncRNA) in nasopharyngeal carcinoma (NPC). We aim to delve into the position of lncRNA HOX antisense intergenic RNA (HOTAIR), together with enhancer of zeste homolog 2 (EZH2), E-cadherin and trimethylation of lysine 27 on histone H3 (H3K27me3) in NPC. METHODS: HOTAIR, EZH2, and E-cadherin expression in NPC tissues and cells were tested. NPC cell biological functions were examined through gain-of and loss-of function assays. The mechanism of lncRNA HOTAIR/E-cadherin/EZH2/H3K27 axis in NPC was decoded. RESULTS: LncRNA HOTAIR and EZH2 were highly expressed in NPC, and E-cadherin was lowly expressed. Down-regulation of HOTAIR or EZH2 inhibited NPC cell progression and tumor growth. HOTAIR recruited histone methylase EZH2 to mediate trimethylation of H3K27 and regulated E-cadherin expression. CONCLUSION: HOTAIR inhibits E-cadherin by stimulating the trimethylation of H3K27 to promote NPC cell progression through recruiting histone methylase EZH2.

[Correlation of single nucleotide polymorphisms of the osteopontin gene with astheozoospermia].

OBJECTIVE: To investigate the correlation of the single nucleotide polymorphisms (SNP) rs1126772, rs117291487, rs11730582, rs142608941 and rs6813526 of the osteopontin (OPN) gene with the risk of asthenozoospermia (AZS). METHODS: We included 135 AZS patients in the AZS group and another 239 fertile men as normal controls. Using the SNaPshot technique, we genotyped the rs1126772, rs117291487, rs11730582, rs142608941 and rs6813526 polymorphisms of the OPN gene in all the subjects and analyzed the correlation of the five SNPs with AZS. RESULTS: The GA genotype and A allele of the OPN gene rs1126772 were found to be correlated with the risk of AZS (GA vs AA: OR = 0.55, 95% CI: 0.35－0.86, P = 0.009; A vs G: OR = 0.64, 95% CI: 0.46－0.89, P = 0.007), and so was the CT genotype and T allele at the RS11730582 locus (CT vs TT: OR = 0.526, 95% CI: 0.34－0.82, P = 0.009; T vs C: OR = 0.60, 95% CI: 0.44－0.83, P = 0.002). Haplotype analysis showed that the AATCT haplotype decreased the risk of AZS (AATCT: OR = 0.61, 95% CI: 0.42－0.88, P = 0.008) . CONCLUSIONS: The polymorphisms of the OPN gene RS1126772 and RS11730582 may reduce the risk of AZS.

Association of interleukin-27 gene polymorphisms with susceptibility to HIV infection and disease progression.

Interleukin-27 (IL-27) gene polymorphisms are linked to infectious disease susceptibility and IL-27 plasma level is associated with HIV infection. Therefore, we aimed to investigate the association between IL-27 polymorphisms and susceptibility to HIV infection and disease progression. A total of 300 patients with HIV infection (48 long-term nonprogressors and 252 typical progressors) and 300 healthy controls were genotyped for three IL-27 polymorphisms, rs17855750, rs181206, rs40837 which were performed by using multiple single nucleotide primer extension technique. Significant association was found between IL-27 rs40837 polymorphisms with susceptibility to HIV infection (AG vs AA: adjusted OR = 1.60, 95% CI, 1.11-2.30, P = 0.012; AG+GG vs AA: adjusted OR = 1.44, 95% CI, 1.02-2.03, P = 0.038) and disease progression (LTNP: AG vs AA: adjusted OR = 2.33, 95% CI, 1.13-4.80, P = 0.021; TP: AG vs AA: adjusted OR = 1.50, 95% CI, 1.04-2.24, P = 0.030). Serum IL-27 levels were significantly lower in cases compared to controls (P < 0.001). There were lower serum IL-27 levels in TPs than in LTNPs (P < 0.001). We further found that LTNPs with rs40837 AG or GG genotype had lower serum IL-27 levels than with AA genotype (P < 0.05). The CD4+ T counts in cases were significantly lower than controls (P < 0.001). In contrast, individuals with rs40837 AG genotype had lower CD4+ T counts than with AA genotype in cases (P < 0.05). In addition, CD4+ T counts in TPs were significantly lower than LTNPs (P < 0.001). IL-27 rs40837 polymorphism might influence the susceptibility to HIV infection and disease progression probably by regulating the level of serum IL-27 or the quantity of CD4+ T.

Exosomal lncRNA HNF1A-AS1 affects cisplatin resistance in cervical cancer cells through regulating microRNA-34b/TUFT1 axis.

BACKGROUND: There is growing evidence of the role of long non-coding RNAs (lncRNAs) in cervical cancer (CC). The objective was to discuss whether exosomal lncRNA HNF1A-AS1 impacted drug resistance in CC via binding to microRNA-34b (miR-34b) and regulating TUFT1 expression. METHODS: The expression of HNF1A-AS1 in normal cervical epithelial cells, cisplatin (DDP)-sensitive cell line (HeLa/S) and DDP-resistant cell line (HeLa/DDP) cells were detected. HeLa/S and HeLa/DDP cells were interfered with HNF1A-AS1 to determine IC50, proliferation, colony formation and apoptosis of CC cells. The exosomes were isolated and identified. Subcellular localization of HNF1A-AS1, expression of miR-34b and TUFT1 in receptor cells were also verified. The binding site between HNF1A-AS1 and miR-34b, together with miR-34b and TUFT1 were confirmed. Tumorigenic ability of cells in nude mice was also detected. RESULTS: HNF1A-AS1 was upregulated in DDP-resistant cell line HeLa/DDP. Silencing HNF1A-AS1 suppressed CC cell proliferation and promoted its apoptosis. HNF1A-AS1 was found to act as a competing endogenous RNA (ceRNA) of miR-34b to promote the expression of TUFT1. Exosomes shuttled HNF1A-AS1 promoted the proliferation and drug resistance of CC cells and inhibited their apoptosis by upregulating the expression of TUFT1 and downregulating miR-34b. Furthermore, suppressed exosomal HNF1A-AS1 in combination with DDP inhibited tumor growth in nude mice. CONCLUSION: Our study provides evidence that CC-secreted exosomes carrying HNF1A-AS1 as a ceRNA of miR-34b to promote the expression of TUFT1, thereby promoting the DDP resistance in CC cells.

Genetic association of promoter in GRP78 gene with nasopharyngeal carcinoma in a Chinese population.

BACKGROUND: Emerging evidences were accumulated to support the view that GRP78 might be associated with multiple types of cancer. Given these, the aim of this study is to investigate the relationship between single nucleotide polymorphisms (SNPs) of GRP78 gene promoter and nasopharyngeal carcinoma (NPC). METHODS: Three SNPs (rs3216733, rs17840761 and rs17840762) in GRR78 promoter were estimated in 422 NPC patients and 452 controls. Genotyping was performed using SNaPshot SNP. Serum GRP78 level was performed by enzyme-linked immunosorbent assay (ELISA). Data were analyzed by SPSS 17.0 software. RESULTS: Significant association between rs3216733 polymorphism and NPC was observed (Cd vs. dd: OR = 0.57, 95% CI 0.43-0.76, P < 0.001; CC vs. dd: OR = 0.62, 95% CI 0.39-0.98, P = 0.043; Cd/CC vs. dd: OR = 0.58, 95% CI 0.44-0.76, P < 0.001; C vs. d OR = 0.70, 95% CI 0.57-0.86, P = 0.001). Additionally, we further found that expression were down-regulated in serum of patients with NPC carrying rs3216733 CC genotype when compared to that of dd genotype (P < 0.001). CONCLUSION: The observations suggest that rs3216733 polymorphism in the GRP78 gene promoter may correlate with NPC susceptibility.

[Asthenozoospermia is not correlated with 3' UTR polymorphisms of the GRP78 gene].

OBJECTIVE: To investigate the correlation of the single nucleotide polymorphisms (SNPs) rs12009, rs1140763 and rs16927997 in the 3'-untranslated region (3'UTR) of the glucose-regulated protein 78 (GRP78) gene with the risk of male asthenozoospermia (AZS). METHODS: We included 400 AZS patients in the AZS group and another 400 fertile men as normal controls. Using the SNaPshot technique, we genotyped the rs12009, rs1140763 and rs16927997 polymorphisms in the 3'UTR of the GRP78 gene in all the male subjects and analyzed the association of the three SNPs with AZS. RESULTS: The percentage of progressively motile sperm was significantly lower in the AZS group than in the normal controls (ï¼»20.09 ± 8.18ï¼½ % vs ï¼»57.16 ± 13.45ï¼½ %, P <0.01). Three genotypes of CC, CT and TT and 2 alleles of C and T were found in rs12009 and rs1140763 of the GRP78 gene, and another three genotypes of GG, GA and AA and two alleles of G and A were observed in rs16927997. There were no statistically significant differences between the control and AZS groups in the frequencies of the C and T alleles in rs12009 (44.3% vs 47.3% and 55.7% vs 52.7%, P >0.05) or rs1140763 (50.0% vs 52.0% and 50.0% vs 48.0%, P >0.05) or those of the G and A alleles in rs16927997 (6.0% vs 4.4% and 94.0% vs 95.6%, P >0.05), nor in the genotypes and allele frequencies of the 3 polymorphisms (P >0.05). Furthermore, three haplotypes of C-C-A, T-C-G and T-T-A were observed in the male subjects but showed no evident correlation between the AZS and normal control groups. CONCLUSIONS: The polymorphisms in the 3'UTR of the GRP78 gene are not correlated with the risk of male asthenozoospermia.

Uncovering the ceRNA network and DNA methylation associated with gene expression in nasopharyngeal carcinoma.

OBJECTIVE: This study aimed to uncover abnormally expressed genes regulated by competitive endogenous RNA (ceRNA) and DNA methylation nasopharyngeal carcinoma and to validate the role of lncRNAs in the ceRNA network on nasopharyngeal carcinoma progression. METHODS: Based on the GSE64634 (mRNA), GSE32960 (miRNA), GSE95166 (lncRNA), and GSE126683 (lncRNA) datasets, we screened differentially expressed mRNAs, miRNAs and lncRNAs in nasopharyngeal carcinoma. A ceRNA network was subsequently constructed. Differentially methylated genes were screened using the GSE62336 dataset. The abnormally expressed genes regulated by both the ceRNA network and DNA methylation were identified. In the ceRNA network, the expression of RP11-545G3.1 lncRNA was validated in nasopharyngeal carcinoma tissues and cells by RT-qPCR. After a knockdown of RP11-545G3.1, the viability, migration, and invasion of CNE-2 and NP69 cells was assessed by CCK-8, wound healing and Transwell assays. RESULTS: This study identified abnormally expressed mRNAs, miRNAs and lncRNAs in nasopharyngeal carcinoma tissues. A ceRNA network was constructed, which contained three lncRNAs, 15 miRNAs and 129 mRNAs. Among the nodes in the PPI network based on the mRNAs in the ceRNA network, HMGA1 was assessed in relation to the overall and disease-free survival of nasopharyngeal carcinoma. We screened two up-regulated genes regulated by the ceRNA network and hypomethylation and 26 down-regulated genes regulated by the ceRNA network and hypermethylation. RP11-545G3.1 was highly expressed in the nasopharyngeal carcinoma tissues and cells. Moreover, the knockdown of RP11-545G3.1 reduced the viability, migration, and invasion of CNE-2 and NP69 cells. CONCLUSION: Our findings uncovered the epigenetic regulation in nasopharyngeal carcinoma and identified the implications of RP11-545G3.1 on the progression of nasopharyngeal carcinoma.

Acrylamide Exposure Destroys the Distribution and Functions of Organelles in Mouse Oocytes.

Acrylamide (ACR) is a common industrial ingredient which is also found in foods that are cooked at high temperatures. ACR has been shown to have multiple toxicities including reproductive toxicity. Previous studies reported that ACR caused oocyte maturation defects through the induction of apoptosis and oxidative stress. In the present study, we showed that ACR exposure affected oocyte organelle functions, which might be the reason for oocyte toxicity. We found that exposure to 5 mM ACR reduced oocyte maturation. ACR caused abnormal mitochondrial distribution away from spindle periphery and reduced mitochondrial membrane potential. Further analysis showed that ACR exposure reduced the fluorescence intensity of Rps3 and abnormal distribution of the endoplasmic reticulum, indicating that ACR affected protein synthesis and modification in mouse oocytes. We found the negative effects of ACR on the distribution of the Golgi apparatus; in addition, fluorescence intensity of vesicle transporter Rab8A decreased, suggesting the decrease in protein transport capacity of oocytes. Furthermore, the simultaneous increase in lysosomes and LAMP2 fluorescence intensity was also observed, suggesting that ACR affected protein degradation in oocytes. In conclusion, our results indicated that ACR exposure disrupted the distribution and functions of organelles, which further affected oocyte developmental competence in mice.

Effects of Lycium barbarum Polysaccharide on Endoplasmic Reticulum Stress and Oxidative Stress in Obese Mice.

BACKGROUND: The incidence of obesity-associated decline in male fertility has increased over the years. Lycium barbarum polysaccharide (LBP), a natural plant polysaccharide extracted from the Chinese herb L. barbarum has shown promising therapeutic effects in overcoming the same. AIM: This study aimed to investigate the protective effect of LBP on the testes of obese mice. METHODS: Following administration of LBP to high-fat diet-induced obese mice for 35 days, serum, sperm, and testis samples were obtained for subsequent experiments. Biochemical analysis and sex hormone content determination were performed to observe changes in glycolipid metabolism and testosterone levels, respectively, in the blood. Hematoxylin and eosin staining were carried out to assess the pathological changes in the testicular tissue. Oxidative stress levels were detected using enzyme-linked immunosorbent assay and expression levels of endoplasmic reticulum stress markers were determined using western blot in the testicular tissue. RESULTS: Our results suggested that LBP reduced glucose levels and insulin resistance, increased testosterone levels and insulin sensitivity, and decreased testicular oxidative stress and pathological damage in obese mice. In addition, LBP down-regulated the expression of p-eIF2α, GRP78, and CHOP in the testicular tissues of obese mice. CONCLUSION: Our results show that LBP is a potential novel drug for preventing male infertility caused by obesity.

Podophyllotoxin Exposure Causes Spindle Defects and DNA Damage-Induced Apoptosis in Mouse Fertilized Oocytes and Early Embryos.

Podophyllotoxin (PPT) is a kind of lignans extracted from the roots and stems of the genus Podophyllum from the tiller family, and it has been widely used in the treatment of condyloma acuminatum, multiple superficial epithelioma in the clinics. However, PPT has been reported to be toxic and can cause liver defects and other organ poisoning. In addition, emerging evidences also indicate that PPT has reproductive toxicity and causes female reproduction disorders. In this study, we used fertilized oocytes and tried to explore the effects of PPT on the early embryonic development with the mouse model. The results showed that exposure to PPT had negative effects on the cleavage of zygotes. Further analysis indicated that PPT could disrupt the organization of spindle and chromosome arrangement at the metaphase of first cleavage. We also found that PPT exposure to the zygotes induced excessive reactive oxygen species (ROS), suggesting the occurrence of oxidative stress. Moreover, in the PPT-exposed embryos, there was positive γH2A.X and Annexin-V signals, indicating that PPT induced embryonic DNA damage and early apoptosis. In conclusion, our results suggested that PPT could affect spindle formation and chromosome alignment during the first cleavage of mouse embryos, and its exposure induced DNA damage-mediated oxidative stress which eventually led to embryonic apoptosis, indicating the toxic effects of PPT on the early embryo development.

[Study on the fluorescence characteristics of the interaction between DNA and two halogenated fluoresceins].

The fluorescence characteristics of the interaction between DNA and two halogen fluoresceins, tetrabromofluorescein (TBF) and tetrachlorotetrabromofluorescein (TTF), were studied. The result showed that for TBF, lamda ex/ lamda em were 518 nm/540 nm, and for TTF, they were 540 nm/560 nm. The intensity of fluorescence of TBF and TTF changed in the presence of DNA. The fluorescence quenching studies indicated that TBF binds to DNA in the groove and the polarization experiments showed that TBF is intercalated into the DNA base pairs. So the comprehensive interaction mode of TBF with DNA may be that TTF is intercalated into DNA base pairs and binds to DNA in the groove. Both the fluorescence quenching studies and the polarization experiments indicated that TTF is intercalated into the DNA base pairs. It was also found that ionic strength could affect the interaction of TBF/TTF and DNA. The binding constants K are 1 x 10(6) L x mol(-1) and 2 x 10(6) L x mol(-1), and the binding site numbers n are 0.62 and 0.16 for TBF and TTF, respectively.

[Study on the interaction of tetrabromofluorescein and DNA].

The interaction of tetrabramofluorescein (TBF) and DNA was studied by the fluorescence and SS-RTP. The effect of the ion strength and ethanol on the fluorescence spectra of TBF in the presence of DNA and absence of DNA was investigated. In addition, the effect of pH on the SS-RTP spectra of TBF was also examined. The quenched constant was 719.74 and 880.22 in the absence and presence of DNA respectively. Finally, the interaction mode between TBF and DNA was discussed from the fluorescence quenched and phosphorescence polarized experiments.

[Study on the interaction of tetraiodofluorescein and bovine serum albumin by fluorimetry].

The binding reaction of tetraiodofluorescein with bovine serum albumin i n aqueous solution was studied by fluorimetry.The binding constant is 2.65 x 10(5) L x mol(-1), the equilibrium constant is 5.7 x 10(4) L x mol(-1), and the binding number is 0.86. The research results of binding model indicate that the hydrophobic force was the main binding force. The presence of BSA can increase the fluorescence intensity of TIF. And the deltaF(F-F0) is linear to the concentration of BSA in a certain range, so the first analytical application was proposed. The linear dynamic range (LDR) was 8 x 10(-7) - 9 x 10(-6) mol x L(-1), the limit of detection (LOD) was 9.88 x 10(-8) mol x L(-1), and the relative standard deviation (RSD) was 3.4%.

Study on the inclusion complexes of cryptotanshinone with beta-cyclodextrin and hydroxypropyl-beta-cyclodextrin.

The inclusion complexes of beta-cyclodextrin (beta-CD) and HP-beta-cyclodextrin (HP-beta-CD) with a kind of tanshinone, cryptotanshinone (CTan) were investigated by using spectrophotometry. Stable inclusion complexes were established in solution and in solid state and were characterized by UV, IR and 1H NMR spectra, respectively. The optimum pH for inclusion is about 7.5. Stoichiometry of the inclusion complex is 1:1. The stabilities of beta-CD and HP-beta-CD to CTan were in the order: beta-CD

Studies on the spectroscopic behavior of cryptotanshinone, tanshinone IIA, and tanshinone I.

A comparative study on the spectroscopic behavior of cryptotanshinone (CTan), tanshinone IIA (Tan IIA), and tanshinone I (Tan I) has been investigated, including UV-Vis absorption, low temperature phosphorescence (LTP), low temperature fluorescence (LTF), paper substrate-room temperature phosphorescence (PS-RTP), paper substrate-room temperature fluorescence (PS-RTF) and fluorescence in liquid (LF). The effect of pH on the luminescence intensity is discussed. Lifetime and polarization of the LTP and RTP have been examined with phosphorescence lifetime in the range of 0.6-0.9s and polarization in the range of 0.10-0.27. Analytical characteristics of LF, PS-RTF and PS-RTP of CTan, Tan IIA, and Tan I have been studied.

[Study on feasible emission control level of air pollutions for cement industry ].

The revised National Emission Standard of Air Pollutions for Cement Industry has been issued, which will be effective for the new enterprises and the existing enterprises on Mar. 1st, 2014 and July 1st, 2015, respectively. In the process of revision, the key technical issues on determination of standard limits was how to determine the feasible emission control level of air pollutions. Feasible emission control requirements were put forward, according to air pollutants emission, technologies, environmental management requirements and foreign standards, etc. The main contents of the revised standard include expanding the scope of application, increasing the pollutants, improving the particulate and NO emissions control level, and increasing special emission limits applied to key areas of air pollutants. The standard will become the gripper of pollution prevention, total emission reduction, structural adjustment and optimization of the layout, and will promote scientific and technical progression for the cement industry.

[A comparative study on domestic and foreign emission standards of air pollutants for cement industry].

The new National Emission Standard of Air Pollutants for Cement Industry (GB 4915-2013) becomes effective on Mar. 1st, 2014. It will play an important role in pollution prevention, total emission reduction, structure adjustment, and layout optimization for cement industry. Based on the research of emission standard in China, U. S., EU and Japan, the similarities and differences in the pollutant projects, control indicators, limits and means of implementation were discussed and advice was proposed, with the purpose to provide a reference for revision of emission standard, and to improve the level of environmental management and pollution control.

[Revision process and thinking of emission standard of air pollutants for cement industry].

The new National Emission Standard of Air Pollutants for Cement Industry (GB 4915-2013) was released recently, which is the third revision since the first release in 1985. This paper reviewed the revision process for the emission standard of air pollutants in cement industry, analyzed the impact of environmental protection situation and management policies changes on the content and form of the standard. The standard formulating principles and several key issues together constitute the base of emission standard, which are not only important to complete the theories and methods of emission standard development, but also important to improve the environmental management and pollution control level.