Specific knockdown of HOXB7 inhibits cutaneous squamous cell carcinoma cell migration and invasion while inducing apoptosis via the Wnt/ß-catenin signaling pathway.

Metastatic cutaneous squamous cell carcinoma (CSCC) is a major cause of death associated with nonmelanoma skin cancer. The involvement of homeobox B7 ( HOXB7) in cancers has been reported. Thus, the current study intends to explore the effect of HOXB7 on CSCC and its relationship with the Wnt/ß-catenin signaling pathway. Initially, microarray-based gene expression profiling of CSCC was performed, and HOXB7 was identified as an upregulated gene based on the microarray data of GSE66359 . Following this, the experimental results indicated that HOXB7 and ß-catenin formed a composite, demonstrating that endogenous HOXB7 binds to ß-catenin. Subsequently, CSCC cells were treated with siRNA against HOXB7 or an inhibitor of the Wnt/ß-catenin signaling pathway to analyze any underlying regulatory mechanism of HOXB7 on the CSCC cells. Tumor growth involving xenografts in nude mice was also observed so as to explore whether or not HOXB7 could regulate subcutaneous tumor growth through in vivo culturing. To investigate the potential effects of HOXB7 on the Wnt/ß-catenin signaling pathway, we determined the expression of HOXB7 and downstream genes of the Wnt/ß-catenin signaling pathway. Notably, siRNA-mediated knockdown of HOXB7 inhibited the activation of the Wnt/ß-catenin signaling pathway, thereby impeding the progression of cell viability, migration, and invasion as well as of the tumor growth, although contrarily facilitating cell apoptosis. Taken together, silencing of the HOXB7 has the mechanism of inactivating the Wnt/ß-catenin signaling pathway, thereby accelerating cell apoptosis and suppressing cell migration and invasion in CSCC, which could provide a candidate target for the CSCC treatment.

Effects of Narrow Band Ultraviolet B on Serum Levels of Vascular Endothelial Growth Factor and Interleukin-8 in Patients with Psoriasis.

The aim of this study was to investigate the effects of narrow band ultraviolet B (NB-UVB) therapy on serum levels of vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8) in patients with psoriasis. Relevant Chinese and English scientific literature databases were searched to identify studies published before November, 2013 that included serum VEGF and IL-8 levels in patients with psoriasis. The studies retrieved from database searches were screened on the basis of predefined selection criteria, and data from finally selected studies were extracted for meta-analysis. Analyses were conducted using STATA 12.0 software. Our systematic search resulted in retrieval of 700 studies (500 studies in Chinese, 200 studies in English), and after a multistep screening process, 13 studies met the inclusion criteria and were enrolled for meta-analysis. The 13 studies contained a combined total of 400 patients with psoriasis and 221 healthy controls. The results of meta-analysis revealed that healthy controls exhibited significantly lower serum level of VEGF, compared to patients with psoriasis before therapy. After NB-UVB, VEGF levels were significantly decreased in patients with psoriasis, as compared to their pretherapy VEGF levels. Although no statistically significant differences were detected in IL-8 serum levels between patients with psoriasis and healthy controls before therapy, after NB-UVB therapy, the serum IL-8 levels in patients with psoriasis were markedly decreased. Corresponding reductions in the psoriasis area and severity index scores of patients with psoriasis were observed after NB-UVB treatment. Our results revealed that NB-UVB therapy significantly decreased the serum levels of VEGF and IL-8 in patients with psoriasis. Furthermore, VEGF and IL-8 levels correlated with disease status, indicating that they are sensitive biomarkers for evaluating the effectiveness of psoriasis therapy.

Whole Transcriptome Analysis Identifies the Taxonomic Status of a New Chinese Native Cattle Breed and Reveals Genes Related to Body Size.

Wandong (WD) cattle has recently been identified as a new Chinese native cattle breed by the National Commission for Livestock and Poultry Genetic Resources. The population size of this breed is less than 10,000. WD cattle and Dabieshan (DB) cattle are sympatric but are raised in different ecological environments, on mountains and plains, respectively, and the body sizes of these two breeds are markedly different. Blood samples were obtained from 8 adult female WD cattle and 7 adult female DB cattle (24 months old). The total RNA was extracted from leukocyte cells, and sequencing experiments were conducted on the Illumina HiSeqTM 4000 platform. After the removal of one outlier sample from the WD cattle breed as determined by principal component analysis (PCA), phylogenetic and population structure analyses indicated that WD and DB cattle formed a distinct Central China cattle group and showed evidence of hybridization between Bos. taurus and Bos. indicus. The immune-regulator CD48 (P = 1.3E-6) was associated with breed-specific traits according to loss-of-function variant enrichment analysis. In addition, 113 differentially expressed genes were identified between the two breeds, many of which are associated with the regulation of body growth, which is the major difference between the two breeds. This study showed that WD cattle belong to the group of hybrids between Bos. Taurus and Bos. indicus, and one novel gene associated with breed traits and multiple differentially expressed genes between these two closely related breeds was identified. The results provide insights into the genetic mechanisms that underlie economically important traits, such as body size, in cattle.

Transcriptional activity of FIGLA, NEUROG2, and EGR1 transcription factors associated with polymorphisms in the proximal regulatory region of GPR54 gene in cattle.

Activity of transcription factors affect synthesis of G-protein coupled receptor 54 (GPR54), an important factor in regulation of initiation of puberty. Expression of the GPR54 gene in cattle is associated with polymorphisms in the proximal regulatory region (PRR) of the GPR54 gene. Transcription resulting in production of GPR54 mRNA transcript occurs as a result of transcription factor (TF) interactions in the PRR. Polymorphisms in the PRR may be associated with extent of activity of these TFs. Folliculogenesis-specific BHLH TF (FIGLA), neurogenin 2 (NEUROG2), and early growth response 1 (EGR1) are important in modulation of ovarian follicle development and neurons synthesizing GnRH, thus, regulating biosynthesis of luteinizing hormone. The aim of this study, therefore, was to assess the transcription-activating potential of binding sites for FIGLA, NEUROG2, and EGR1 TFs in the GPR54 promoter of cattle. Two luciferase-based promoters, ATC and CCT, which contain three single nucleotide polymorphisms (SNPs), A/C-794, T/C-663, and C/T-601, in the GPR54 PRR, were analyzed to evaluate gene expression and activation of different promoters by FIGLA, NEUROG2, and EGR1. The FIGLA induced GPR54 transcription through the CCT, whereas NEUROG2 and EGR1 induced GPR54 transcription through the ATC promoter-binding site. The CCT-activating effects of FIGLA were greater (2.56-fold) than the ATC-activating effects (P < 0.05). The ATC-activating effects of NEUROG2 and EGR1 were markedly greater (12.91- and 8.41-fold; P < 0.01) than CCT-activating effects. The polymorphisms, CCT and ATC, of the cattle GPR54 affect the activity of transcription factors, therefore, have an important effect on production of GPR54 mRNA transcript.

[Polymorphic and linkage analysis of microsatellite BMS2508 and FecB gene in sheep].

Genetic polymorphisms of microsatellite locus BMS2508, which was closely linked to the ovine fecundity gene FecB, were detected in prolific (Small Tail Han sheep) and non-prolific breeds of sheep (Texel, Dorset and Chinese Merino). The linkage disequilibrium between microsatellite locus BMS2508 and FecB gene of Small Tail Han sheep was also analyzed. There was the same mutation (A746G) of BMPR-IB gene in Small Tail Han sheep as that of FecB in Booroola Merino ewes, but the FecB mutation was absent in Texel, Dorset and Chinese Merino sheep. The genotype frequencies of BB, B+ and ++ were 0.485, 0.398 and 0.117 in Small Tail Han sheep, respectively. There were eight alleles varied from 94 bp to 116 bp and 15 genotypes detected at BMS2508 locus in four sheep breeds totally 438 individual. The preponderant allele was 100 bp, 94 bp, 94 bp, 112 bp, 100 bp, 100 bp, 112 bp, and the frequency was 0.453, 0.544, 0.802, 0.475, 0.483, 0.439, 0.389 in Small Tail Han (n=307), Texel (n=45), Dorset (n=46), Chinese Merino (n=40), and BB group (n=149), B+ group (n=122), ++ group (n=36) from Small Tail Han, respectively. In Small Tail Han sheep, linkage analysis indicated that there was certain linkage disequilibrium between 100 bp allele of microsatellite BMS2508 and B allele of FecB gene (D' =0.408), and certain linkage disequilibrium between 110 bp and 114 bp alleles of microsatellite BMS2508 and + allele of FecB gene (D'=0.513).

Inhibitory effect of microRNA-154 targeting WHSC1 on cell proliferation of human skin squamous cell carcinoma through mediating the P53 signaling pathway.

OBJECTIVE: Skin squamous cell carcinoma (SCC) is a common, morbid, and frequently lethal malignancy and ranks as the sixth most deadly cancer worldwide. Hence, this study aims to explore the effect of microRNA-154 (miR-154) targeting WHSC1 on proliferation and apoptosis of SCC cells via the P53 signaling pathway. METHODS: The targeting relationship between WHSC1 and miR-154 was validated using dual-luciferase reporter assay. Normal human epidermal keratinocytes (NHEK) were included, and SCC A431 and SCC-15 cell lines were cultured and transfected with miR-154 mimic, miR-154 inhibitor or siRNA-WHSC1. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) and western blot analysis were used for the miR-154 expression and levels of WHSC1, P53 signaling pathway- and apoptosis-related genes. MTT assay and flow cytometry were applied to determine the cell viability and apoptosis. RESULTS: WHSC1 is a target gene of miR-154. MiR-154 negatively regulated WHSC1 expression and inhibited the activation of P53 signaling pathway. In response to miR-154 mimic or siRNA-WHSC1, SCC A431 and SCC-15 cell lines exhibited increased expression of P73, P16 and Bax, decreased expression of WHSC1, P53, c-myc and Bcl-2, as well as attenuated cell viability and enhanced cell apoptosis. The treatment of miR-154 inhibitor reversed the tendency. CONCLUSION: These results demonstrate that up-regulation of miR-154 inhibits proliferation and induces apoptosis of human skin SCC cells by down-regulating WHSC1 and blocking the P53 signaling pathway.

[Methodology of mapping quantitative trait loci for discrete traits using maximum likelihood].

The maximum likelihood method was used to compare the efficiency of interval mapping with either the threshold model or the linear model. The irfluencing factors of quantitative trait loci (QTL) detection efficiency (e.g. QTL effect, heritability and incidence of categories) were simulated in our study. Daughter design with multiple families was applied, and the number of segregating population was 500. The results showed that the threshold model was superior in terms of parameter estimation. It was a more efficient and accurate model of QTL mapping for discrete traits. In addition, the accuracy of QTL mapping depended on the effect of the putative QTL, the value of heritability and incidence directly. With an increase of QTL effect, heritability and incidence of categories, the accuracy of QTL mapping improved correspondingly.

Effects of nuclear factor-κB and ERK signaling transduction pathway inhibitors on human melanoma cell proliferation in vitro.

The present study aimed to investigate the effects of blocking nuclear factor (NF)-κB and/or extracellular signal-regulated kinase (ERK) signaling pathways on proliferation and apoptosis of melanoma cells in vitro. A375 Human melanoma cells were treated with U0126 (ERK signaling pathway inhibitor) and BMS-345541 (NF-κB inhibitor), alone or in combination. At 12, 24 and 48 h after treatment, cell viability was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, cell cycle progression and apoptosis were evaluated by flow cytometry, and Bcl-2 protein content was determined by western blot analysis. BMS-345541 and U0126 significantly inhibited A375 cell proliferation in a dose- and time-dependent manner (P<0.01). The rate of proliferation inhibition at 24 h was 35.41±1.38% for BMS-345541 alone, 30.64±2.86% for U0126 alone, and 77.27±2.70% for BMS-345541 and U0126 in combination. The difference between combination and single treatment was significantly different (P<0.01). The proportion of cells in S phase was 14.20, 18.40 and 22.64% following treatment with BMS-345541, U0126, and BMS-345541 and U0126 in combination, respectively; these values were all significantly reduced compared with the untreated control group (P<0.01). The apoptosis rate was 24.98±1.03% in the BMS-345541 group, 13.96±0.96% in the U0126 group and 38.91±1.46% in the combination group; all significantly increased compared with the control group (P<0.01). Bcl-2 protein content in A375 cells was significantly increased following treatment with BMS-345541 and U0126, alone or in combination, when compared with the untreated control group (P<0.01). Therefore, NF-κB and ERK signaling pathway inhibitors may serve as potential therapeutic targets for melanoma.

[Study on high-risk behaviour and suicide associated risk factors related to HIV/AIDS among gay or bisexual men].

OBJECTIVE: Characteristics on AIDS high-risk behaviors in gay or bisexual men with suicide ideas were explored and analyzed. METHODS: A cross-sectional survey was conducted with the snowball sampling method adopted. Subjects with suicide ideas were collected from responses to the valid questionnaires and subjects with no suicide ideas were collected from the age comparable men. RESULTS: The overall rate of gays or bisexuals with suicide ideas was 20.2% in this survey. The attitude for homogeneity and marital status among the unmarried was more than that among the comparable group (P < 0.05). The rate of AIDS high-risk behaviors as same-sex sexual harassment, bleeding during sexual intercourse in the last year, coitus with unfamiliar same-sex partners in cities, suffering from adult same-sex sexual abuse before the age of 16, having had sexual abuse and abusive behavior, having had active or passive anal kiss, having had active or passive coitus with fingers, alcohol consumption weekly at least once or more, hurt by gays because of attitude and/or same-sex sexual activity and hurt by heterosexual men because of attitude and/or same-sex sexual activity were significantly higher in gays and bisexual men with suicide ideas than those without (P < 0.05). Data from multivariate logistic regression models suggested that harm from gays (Waldχ(2) = 6.637, P = 0.010) and heterosexual men (Waldχ(2) = 5.835, P = 0.016) due to attitude on homosexual activity appear to be the risk factors causing the suicide ideas. CONCLUSION: Reducing the social discrimination and harm towards gays and bisexual men could reduce the occurrence of the suicide ideas and have a positive effect on curbing the prevalence of AIDS.