Decreased intranuclear cardiac troponin I impairs cardiac autophagy through FOS/ATG5 in ageing hearts.

In our previous study, intranuclear cardiac troponin I (cTnI) may function as a co-factor of Yin Yang 1(YY1). Here, we aimed to explore the role of intranuclear cTnI in ageing hearts. Nuclear translocation of cTnI was demonstrated using Western blot and immunofluorescence. The potential nuclear localization sequences (NLSs) of cTnI were predicted by a web server and then verified in 293T cells by putative NLS-eGFP-GST and NLS-mutant transfection. The ratio of Nuclear cTnI/ Total cTnI (Nu/T) decreased significantly in ageing hearts, accompanied with ATG5-decline-related impaired cardiac autophagy. RNA sequencing was performed in cTnI knockout hearts. The differential expressed genes (DEGs) were analysed by overlapping with YY1 ChIP-sequencing data. cTnI gain and loss experiments in vitro determined those filtered DEGs' expression levels. A strong correlation was found between expression patterns cTnI and FOS. Using ChIP-q-PCR, we demonstrated that specific binding DNA sequences of cTnI were enriched in the FOS promoter -299 to -157 region. It was further verified that pcDNA3.1 (-)-cTnI could increase the promoter activity of FOS by using luciferase report assay. At last, we found that FOS can regulate the ATG5 (autophagy-related gene 5) gene by using a luciferase report assay. Taken together, our results indicate that decreased intranuclear cTnI in ageing hearts may cause impaired cardiac autophagy through the FOS/ATG5 pathway.

Sodium butyrate inhibits oral squamous cell carcinoma proliferation and invasion by regulating the HDAC1/HSPB7 axis.

The study was performed to ascertain the mechanism of sodium butyrate (NaB) mediating the proliferative and invasive properties of oral squamous cell carcinoma (OSCC) cells. The cell proliferative, migrating, and invasive potentials were detected by CCK-8, colony formation, EdU, and Transwell assays. The expression of proliferation- and invasion-related proteins, HDAC1, and HSPB7 in OSCC cells were evaluated by western blot. Immunofluorescence was also performed to evaluate the HDAC1 expression. The enrichment of histone deacetylase HDAC1 in the promoter region of HSPB7 was assessed by the ChIP assay. In vivo growth of OSCC cells was measured by tumorigenesis in nude mice (n=18). The t-test was employed for comparisons of data between the two groups. One-way ANOVA was utilized for comparisons of data among multiple groups, and repeated-measures ANOVA for comparisons of data at different time points among groups, followed by Bonferroni post-hoc test. The data showed that HDAC1 expression was highly upregulated in OSCC cells compared to human normal oral keratinocytes (HNOKs) (p<0.0001), and NaB diminished the HDAC1 expression in OSCC cells. NaB restricted OSCC cell proliferative, migrating, and invasive capabilities by downregulating HDAC1. HSPB7 expression was downregulated in OSCC cells versus HNOKs (p<0.0001). HDAC1 inversely orchestrated the HSPB7 expression in OSCC cells through histone deacetylation modification, and NaB augmented the HSPB7 expression by inhibiting HDAC1. Moreover, NaB inhibited OSCC cell growth in vivo by elevating HSPB7 levels through the HDAC1 repression. In conclusion, NaB restrained cell proliferation and invasion in OSCC cells via HSPB7 upregulation by decreasing the HDAC1 expression.

Does the incorporation of strontium into calcium phosphate improve bone repair? A meta-analysis.

BACKGROUND: The application of calcium phosphate (CaP)-based bone substitutes plays an important role in periodontal regeneration, implant dentistry and alveolar bone reconstruction. The incorporation of strontium (Sr) into CaP-based bone substitutes appears to improve their biological properties, but the reported in vivo bone repair performance is inconsistent among studies. Herein, we conducted a systematic review and meta-analysis to investigate the in vivo performance of Sr-doped materials. METHODS: We searched PubMed, EMBASE (via OVIDSP), and reference lists to identify relevant animal studies. The search, study selection, and data extraction were performed independently by two investigators. Meta-analyses and sub-group analyses were conducted using Revman version 5.4.1. The heterogeneity between studies were assessed by I2. Publication bias was investigated through a funnel plot. RESULTS: Thirty-five studies were finally enrolled, of which 16 articles that reported on new bone formation (NBF) were included in the meta-analysis, covering 31 comparisons and 445 defects. The overall effect for NBF was 2.25 (95% CI 1.61-2.90, p < 0.00001, I2 = 80%). Eight comparisons from 6 studies reported the outcomes of bone volume/tissue volume (BV/TV), with an overall effect of 1.42 (95% CI 0.65-2.18, p = 0.0003, I2 = 75%). Fourteen comparisons reported on the material remaining (RM), with the overall effect being -2.26 (95% CI - 4.02 to - 0.50, p = 0.0009, I2 = 86%). CONCLUSIONS: Our study revealed that Sr-doped calcium phosphate bone substitutes improved in vivo performance of bone repair. However, more studies are also recommended to further verify this conclusion.

[Correlation between high-risk TP53 mutation and extracapsular spread in oral squamous cell carcinoma].

PURPOSE: To investigate the correlation between high-risk tumor protein p53 (TP53) mutation and extracapsular spread(ECS) in oral squamous cell carcinoma (OSCC). METHODS: The data of 88 OSCC patients admitted to Gansu Provincial People's Hospital from January 2013 to January 2016 were retrospectively analyzed. The patients were divided into non-lymph node metastasis group (A1), lymph node metastasis without ECS group (A2), lymph node metastasis with ECS group(A3) according to the presence or absence of lymph node metastasis. The deletion of exon 5-8 of TP53 gene in primary and metastatic lesions was detected. The correlation of ECS with disease-free survival(DFS), overall survival(OS) rate and TP53 mutation were determined, and single factor analysis of ECS was performed. The data were analyzed by SPSS 20.0 software package. RESULTS: TP53 (5) homozygote deletion was found in all three groups. TP53 (6) homozygous deletion was found in group A3 (10/30). No homozygous deletion of TP53(7) was found in three groups.Homozygous deletion of TP53 (8) was found in group A2(7/42), group A3(10/30) and group A3(10/60).The 1-year and 3-year DFS rates were 75.00% and 70.83% in group A1, 70.59% and 58.82% in group A2, 46.67% and 40.00% in group A3, with significant difference(P<0.05). The 1-year and 3-year OS rates were 83.33% and 75.00% in group A1, 73.53% and 50.00% in group A2, 46.67% and 40.00% in group A3 , with significant difference(P<0.05). In group A3, there were 4 cases (13.33%) of low-risk TP53 mutation (LR), 12 cases (40.00%) of high-risk TP53 mutation (HR), 4 cases(13.33%) of wild-type TP53 mutation (Wt), and 10 cases(33.33%) of other mutations. HR mutation and other mutations occurred more frequently than other types(P<0.05). Smoking, primary lesion size, wild type/low risk and high risk/others were correlated with ECS(P<0.05). CONCLUSIONS: ECS is an important marker of DFS and OS in OSCC patients, and high-risk mutations were common in ECS, indicating a certain correlation between high-risk mutations of TP53 and ECS in OSCC.

Macrophage migration inhibitory factor promotes tumor aggressiveness of esophageal squamous cell carcinoma via activation of Akt and inactivation of GSK3ß.

The pleiotropic pro-inflammatory cytokine, macrophage migration inhibitory factor (MIF), represents an important link between chronic inflammation and tumorigenesis. Although accumulating evidence demonstrates that MIF overexpression is implicated in the development and progression of multiple cancers, including esophageal squamous cell carcinoma (ESCC), the molecular mechanisms underlying its tumor-promoting roles in ESCC remain unclear. In the present study, we observed that MIF is overexpressed in ESCC and correlated significantly with lymph node metastasis, advanced clinical stage, and poor survival of ESCC. MIF knockdown attenuated the proliferation, migration, and invasion of ESCC cells in vitro and in vivo. Moreover, blockage of MIF expression decreased the activation of the Akt, MEK/ERK, and NF-κB pathways and enhanced sensitivity to apoptosis. Meanwhile, repression of MIF expression resulted in activation of glycogen synthase kinase 3 beta (GSK3ß) and subsequent decrease of active ß-catenin, as well as its downstream targets including cyclin D1, matrix metalloproteinase (MMP)-7, c-myc, and c-Jun. Collectively, our results provided mechanistic insights into the tumor-promoting role of MIF in ESCC, and suggested that MIF represents a potential therapeutic target for treatment of ESCC.

Integrative topological analysis of mass spectrometry data reveals molecular features with clinical relevance in esophageal squamous cell carcinoma.

Combining MS-based proteomic data with network and topological features of such network would identify more clinically relevant molecules and meaningfully expand the repertoire of proteins derived from MS analysis. The integrative topological indexes representing 95.96% information of seven individual topological measures of node proteins were calculated within a protein-protein interaction (PPI) network, built using 244 differentially expressed proteins (DEPs) identified by iTRAQ 2D-LC-MS/MS. Compared with DEPs, differentially expressed genes (DEGs) and comprehensive features (CFs), structurally dominant nodes (SDNs) based on integrative topological index distribution produced comparable classification performance in three different clinical settings using five independent gene expression data sets. The signature molecules of SDN-based classifier for distinction of early from late clinical TNM stages were enriched in biological traits of protein synthesis, intracellular localization and ribosome biogenesis, which suggests that ribosome biogenesis represents a promising therapeutic target for treating ESCC. In addition, ITGB1 expression selected exclusively by integrative topological measures correlated with clinical stages and prognosis, which was further validated with two independent cohorts of ESCC samples. Thus the integrative topological analysis of PPI networks proposed in this study provides an alternative approach to identify potential biomarkers and therapeutic targets from MS/MS data with functional insights in ESCC.

Downregulation of 14-3-3σ correlates with multistage carcinogenesis and poor prognosis of esophageal squamous cell carcinoma.

AIMS: The asymptomatic nature of early-stage esophageal squamous cell carcinoma (ESCC) results in late presentation and consequent dismal prognosis This study characterized 14-3-3σ protein expression in the multi-stage development of ESCC and determined its correlation with clinical features and prognosis. MATERIALS AND METHODS: Western blot was used to examine 14-3-3σ protein expression in normal esophageal epithelium (NEE), low grade intraepithelial neoplasia (LGIN), high grade intraepithelial neoplasia (HGIN), ESCC of TNM I to IV stage and various esophageal epithelial cell lines with different biological behavior. Immunohistochemistry was used to estimate 14-3-3σ protein in 110 biopsy samples of NEE, LGIN or HGIN and in 168 ESCC samples all of whom had follow-up data. Support vector machine (SVM) was used to develop a classifier for prognosis. RESULTS: 14-3-3σ decreased progressively from NEE to LGIN, to HGIN, and to ESCC. Chemoresistant sub-lines of EC9706/PTX and EC9706/CDDP showed high expression of 14-3-3σ protein compared with non-chemoresistant ESCC cell lines and immortalized NEC. Furthermore, the downregulation of 14-3-3σ correlated significantly with histological grade (P = 0.000) and worse prognosis (P = 0.004). Multivariate Cox regression analysis indicated that 14-3-3σ protein (P = 0.016) and T stage (P = 0.000) were independent prognostic factors for ESCC. The SVM ESCC classifier comprising sex, age, T stage, histological grade, lymph node metastasis, clinical stage and 14-3-3σ, distinguished significantly lower- and higher-risk ESCC patients (91.67% vs. 3.62%, P = 0.000). CONCLUSIONS: Downregulation of 14-3-3σ arises early in the development of ESCC and predicts poor survival, suggesting that 14-3-3σ may be a biomarker for early detection of high-risk subjects and diagnosis of ESCC. Our seven-feature SVM classifier for ESCC prognosis may help to inform clinical decisions and tailor individual therapy.

The CRISPLD2 gene is involved in cleft lip and/or cleft palate in a Chinese population.

BACKGROUND: Nonsyndromic cleft lip and/or cleft palate (NSCLP) are common congenital anomalies in humans, the etiologies of which are complex and associated with both genetic and environmental factors. Previous data suggested single nucleotide polymorphisms (SNPs) of rs1546124, rs4783099, and rs16974880 of the CRISPLD2 gene were associated with an increased risk of NSCLP; however, subsequent studies have yielded conflicting results. This study aims to evaluate the associations of the aforementioned polymorphisms with NSCLP in a Northwestern Chinese population. METHODS: Three CRISPLD2 SNPs were genotyped in a case-control study (n = 907), including 444 NSCLP patients and 463 healthy individuals, using polymerase chain reaction-denaturing high-performance liquid chromatography (PCR-DHPLC). RESULTS: The genotype and allele frequencies of rs1546124 (odds ratio [OR], 2.30; 95% confidence interval [CI], 1.58-3.34; p = 1 × 10(-5) ) and rs4783099 (OR, 0.73; 95% CI, 0.54-1.00; p = 0.05) were different in NSCLP patients compared with controls. Furthermore, the CC genotype at rs1546124 was associated with increased risk for cleft lip with or without cleft palate (CL/P; OR, 2.11; 95% CI, 1.41-3.15; p(correct) = 1.5 × 10(-4) ) and for cleft palate only (CPO; OR, 2.93; 95% CI, 1.69-5.07; p(correct) = 5.4 × 10(-4) ), whereas the T allele of rs4783099 was associated with decreased risk for CPO. Further gender stratification showed that the statistical association of these two loci is mainly in the male patients, and not in female patients. CONCLUSION: Our results suggest that the CRISPLD2 gene contributes to the etiology of NSCLP in the Northwestern Chinese population. SNP rs1546124 is significantly related to NSCLP, associated with both CL/P and CPO groups, and SNP rs4783099 is significantly associated with CPO.

[Risk assessment of nitrogen and phosphorus export in upper reach of Daliao river watershed].

With the development of point pollution control, non-point source pollution has become an important environmental and water quality management problem. Land cover composition across a watershed is a dominant factor in controlling the amount of nitrogen and phosphorus exported from a watershed. Generally, urban and agricultural land covers are considered as principal sources of excess loads of nitrogen (N) and phosphorous (P) in receiving waters. A well developed literature of nutrient export coefficients by land-cover class was used to model the risk of equaling or exceeding specified levels of nutrient export in drainage basin of Dahuofang Reservoir. The model was applied to about 513 comparatively small watersheds mapped for the drainage basin of Dahuofang Reservoir for environmental analysis and planning. The results suggest that the probabilities of risk are 19.31% and 8.95% for N and P nutrient respectively. As the spatial distribution concerned, risk estimates generally differed with different slope degrees and districts. Risk estimates generally increased from the places where lies near the rivers to the places where are far from the river, but numerous areas of high variability were evident.

[Contents, contamination and geochemical characteristics of metals in the sediment from Songhua River].

Geochemical behavior of major and trace elements in the sediment of the Songhua River was studied. 39 bottom sediment samples were collected from the Songhua River and total contents of As, Cd, Co, Cr, Cu, Hg, Zn, Ni, Pb, Sb, Sc, V, Mn, Ti, Al, Fe, Mg, Ca, Na and K in these samples were measured. The objectives of this study were to identify the contents and spatial variations of these metals in the sediment, investigate geochemical relationships among the elements, and develop baseline relationships between 9 trace metals and conservative reference element Sc to quantify the contamination of these 9 elements. Results indicated that the concentrations of toxic trace metals, such as As, Cd, Co, Cr, Cu, Hg, Zn, Ni, Pb, and V in the sediment, were 2.7-11.5, 0.05-1.38, 4.8-14.7, 15.9-78.9, 2.4-75.4, 0.01-1.27, 21.8-403.1, 6.2-35.8, 12.6-124.4, and 22.1-108.0 mg/kg, respectively. Due to the input of anthropogenic sources, temporal and spatial variation of Cd, Cu, Hg, Zn, and Pb contents in the sediment was higher than that of major elements. In addition, correlations between these metals and their mineral matrix elements such as Fe, Mg, and Sc were decreased. The baseline relationships between general metal contaminants and Sc were developed, with high correlation coefficients for Co, Cr, Cu, Zn, Ni, Pb, and V and relatively low correlation coefficients for As and Cd. These baseline relationships provide a way to quantitatively evaluate the sediment contamination by these metals. Generally, sediment contamination of the Songhua River by trace metals was less than that of the Zhujiang River and the Changjiang River, and similar to that of the Huanghe River.

[Regional environment risk assessment and probability distribution of topsoil PAHs in Tianjin area].

Based on some foreign standards of soil PAHs assessment, regional environment risk assessment of topsoil PAHs in Tianjin Area was discussed using the method of indicator Kriging. The results using different standards are also compared. It is shown that Bap is the only one component which exceeds the preliminary remedial goals and risk based concentration of U.S. in Tianjin area and the regions of exceeding standards are about 8.12% and 2.34% respectively. Most components of soil PAHs in Tianjin area are exceed the standard of Holland. The regions of exceeding standard are mostly 90% except the components of Ant (5.26%) and Bas (68.42%). Otherwise, the regions of exceeding Canada standard are mostly under 5% except Nap (97.89%), Phe (56.84%) and Pyr (47.65%). Based on the Soil Plan Zealand of Denmark, Nap is the only one component which exceeds the standard in Tianjin area and the region is about 9.26%. Comparing these risk distribution maps and spatial distribution maps of PAHs, it is found that the areas with high risk are mainly fallen in the areas with high concentrations of PAHs. The probability maps of PAHs risk are helpful for regional environmental management and planning.

[Effect of land use/cover change on pollution load of non-point source in upper reach of Yangtze River Basin].

On the basis of other scholars' researches, utilizes export coefficient model, adopts RS and GIS techniques, estimates the non-point source (NPS) pollution load of upper reach of Yangtze River Basin, and simulates its special distribution. The results indicates that the total nitrogen load caused by land use drop from 1.23 x 10(6) tons in 1970s' to 1.16 x 10(6) tons in 2000 on the premise of taking no account of basin loss. It reduced year by year basically in the past several decades and so did TP load which decreased from 3.7 x 10(4) tons in 1970s' to 3.5 x 10(4) tons in 2000. As far as province, land use and water system are considered, Sichuan province, crop land grass, Jinsha river and Jialing river are important contributories of NPS pollution load in study area. Intensity analysis shows the region of Chongqing municipality and the watershed of Jialing River are two highest NPS pollution load areas, and these areas should be gained more attention in the future. Using the method put forward in this paper, NPS pollution space simulation is carried out in large scale basin such as upper reach of Yangtze River Basin precisely basically.

[Multi-scale spatial relations of PAHs and soil properties in Tianjin area].

Polycyclic aromatic hydrocarbons(PAHs) in soil have serious latent danger to our human health. Based on the sampling data, the spatial scale-dependent correlations of PAHs content and some soil properties are analyzed by Factorial Kriging for top soil in Tianjin area. Based on 1 88 topsoil samples in Tianjin area, pH, TOC, CLAY and sum of 16prior PAHs are tested. Resultsshow that significant difference on spatial correlations at different spatial scales between the PAHs content and soil properties such as pH, TOC and CLAY existed in the studies area.