A Novel missense mutation of COL2A1 gene in a large family with stickler syndrome type I.

Stickler syndrome type I (STL1, MIM 108300) is characterized by ocular, auditory, skeletal and orofacial manifestations. Nonsyndromic ocular STL1 (MIM 609508) characterized by predominantly ocular features is a subgroup of STL1, and it is inherited in an autosomal dominant manner. In this study, a novel variant c.T100>C (p.Cys34Arg) in COL2A1 related to a large nonsyndromic ocular STL1 family was identified through Exome sequencing (ES). Bioinformatics analysis indicated that the variant site was highly conserved and the pathogenic mechanism of this variant may involve in affected structure of chordin-like cysteine-rich (CR) repeats of ColIIA. Minigene assay indicated that this variant did not change alternative splicing of exon2 of COL2A1. Moreover, the nonsyndromic ocular STL1 family with 16 affected members showed phenotype variability and certain male gender trend. None of the family members had hearing loss. Our findings would expand the knowledge of the COL2A1 mutation spectrum, and phenotype variability associated with nonsyndromic ocular STL1. Search for genetic modifiers and related molecular pathways leading to the phenotype variation warrants further studies.

Toxicological Mechanism of Individual Susceptibility to Formaldehyde-Induced Respiratory Effects.

Understanding the mechanisms of individual susceptibility to exposure to environmental pollutants has been a challenge in health risk assessment. Here, an integrated approach combining a CRISPR screen in human cells and epidemiological analysis was developed to identify the individual susceptibility to the adverse health effects of air pollutants by taking formaldehyde (FA) and the associated chronic obstructive pulmonary disease (COPD) as a case study. Among the primary hits of CRISPR screening of FA in human A549 cells, HTR4 was the only gene genetically associated with COPD susceptibility in global populations. However, the association between HTR4 and FA-induced respiratory toxicity is unknown in the literature. Adverse outcome pathway (AOP) network analysis of CRISPR screen hits provided a potential mechanistic link between activation of HTR4 (molecular initiating event) and FA-induced lung injury (adverse outcome). Systematic toxicology tests (in vitro and animal experiments) were conducted to reveal the HTR4-involved biological mechanisms underlying the susceptibility to adverse health effects of FA. Functionality and enhanced expression of HTR4 were required for susceptibility to FA-induced lung injury, and FA-induced epigenetic changes could result in enhanced expression of HTR4. Specific epigenetic and genetic characteristics of HTR4 were associated with the progression and prevalence of COPD, respectively, and these genetic risk factors for COPD could be potential biomarkers of individual susceptibility to adverse respiratory effects of FA. These biomarkers could be of great significance for defining subpopulations susceptible to exposure to FA and reducing uncertainty in the next-generation health risk assessment of air pollutants. Our study delineated a novel toxicological pathway mediated by HTR4 in FA-induced lung injury, which could provide a mechanistic understanding of the potential biomarkers of individual susceptibility to adverse respiratory effects of FA.

CRISPR screen identified that UGT1A9 was required for bisphenols-induced mitochondria dyshomeostasis.

Exposure to bisphenols chemicals could cause various adverse health effects, including non-alcoholic fatty liver disease (NAFLD), which have been associated with cellular mitochondria stress. However, the biological mechanism underlying the mitochondria stress-mediated cell death by bisphenols was poorly understood. Here, CRISPR screens were performed to identify the critical genes which were involved in cell death caused by exposure to four bisphenols (BPA, BPB, BPE and BPS). Results of CRISPR screens showed that UGT1A9 was the primary genetic factor facilitating cell death induced by all of the four bisphenols. Systematic toxicological tests demonstrated that UGT1A9 was required for BPA-induced mitochondria dyshomeostasis in vitro and in vivo, and UGT1A9-mediated mitochondria dyshomeostasis was an important cause of facilitating cell death. Liver injury caused by exposure to BPA in wild-type mice was accompanied with suppression of mitophagy and increased expression of C-Caspase 3, but UGT1A9 knockout attenuated these adverse effects induced by BPA. Finally, molecular epidemiology analysis suggested that the five genetic variants of UGT1A9 could be potential genetic risk factors of NAFLD when people were exposed to BPA. The biological mechanism uncovered here provided mechanistic evidence for identification of susceptible populations of liver injury associated with exposure to BPA.

Evaluation of dioxin induced transcriptomic responses in a 3D human liver microtissue model.

Three-dimensional human liver microtissue model provides a promising method for predicting the human hepatotoxicity of environmental chemicals. However, the dynamics of transcriptional responses of 3D human liver microtissue model to dioxins exposure remain unclear. Herein, time-series transcriptomic analysis was used to characterize modulation of gene expression over 14 days in 3D human liver microtissues exposed to 2,3,7,8-tetra-chlorodibenzo-p-dioxin (TCDD, 31 nM, 10 ng/ml). Changes in gene expression and modulation of biological pathways were evaluated at several time points. The results showed that microtissues stably expressed genes related to toxicological pathways (e.g. highly of genes involved in external stimuli and maintenance of cell homeostasis pathways) during the 14-day culture period. Furthermore, a weekly phenomenon pattern was observed for the number of the differentially expressed genes in microtissues exposed to TCDD at each time point. TCDD led to an induction of genes involved in cell cycle regulation at day three. Metabolic pathways were the main significantly induced pathways during the subsequent days, with the immune/inflammatory response enriched on the fifth day, and the cellular response to DNA damage was identified at the end of the exposure. Finally, relevant transcription patterns identified in microtissues were compared with published data on rodent and human cell-line studies to elucidate potential species-specific responses to TCDD over time. Cell development and cytochrome P450 pathway were mainly affected after a 3-day exposure, with the DNA damage response identified at the end of exposure in the human microtissue system but not in mouse/rat primary hepatocytes models. Overall, the 3D human liver microtissue model is a valuable tool to predict the toxic effects of environmental chemicals with a relatively long exposure.

Cross-Model Comparison of Transcriptomic Dose-Response of Short-Chain Chlorinated Paraffins.

Short-chain chlorinated paraffins (SCCPs) have attracted attention because of their toxicological potential in humans and wildlife at environmentally relevant doses. However, limited information is available regarding mechanistic differences across species in terms of the biological pathways that are impacted by SCCP exposure. Here, a concentration-dependent reduced human transcriptome (RHT) approach was conducted to evaluate 15 SCCPs in HepG2 cells and compared with our previous results using a reduced zebrafish transcriptome (RZT) approach in zebrafish embryos (ZFEs). Generally, SCCPs induced a broader suite of biological pathways in ZFEs than HepG2 cells, and all of the 15 SCCPs were more potent in HepG2 cells compared to ZFEs. Despite these general differences, the transcriptional potency of SCCPs in both model systems showed a significant linear relationship (p = 0.0017, r2 = 0.57), and the average ratios of transcriptional potency for each SCCP in RZT to that in RHT were ∼100,000. C10H14Cl8 was the most potent SCCP, while C10H17Cl5 was the least potent in both ZFEs and HepG2 cells. An adverse outcome pathway network-based analysis demonstrated model-specific responses, such as xenobiotic metabolism that may be mediated by different nuclear receptor-mediated pathways between HepG2 cells (e.g., CAR and AhR activation) and ZFEs (e.g., PXR activation). Moreover, induced transcriptional changes in ZFEs associated with pathways and molecular initiating events (e.g., activation of nicotinic acetylcholine receptor) suggest that SCCPs may disrupt neural development processes. The cross-model comparison of concentration-dependent transcriptomics represents a promising approach to assess and prioritize SCCPs.

Benchmarking Water Quality from Wastewater to Drinking Waters Using Reduced Transcriptome of Human Cells.

One of the major challenges in environmental science is monitoring and assessing the risk of complex environmental mixtures. In vitro bioassays with limited key toxicological end points have been shown to be suitable to evaluate mixtures of organic pollutants in wastewater and recycled water. Omics approaches such as transcriptomics can monitor biological effects at the genome scale. However, few studies have applied omics approach in the assessment of mixtures of organic micropollutants. Here, an omics approach was developed for profiling bioactivity of 10 water samples ranging from wastewater to drinking water in human cells by a reduced human transcriptome (RHT) approach and dose-response modeling. Transcriptional expression of 1200 selected genes were measured by an Ampliseq technology in two cell lines, HepG2 and MCF7, that were exposed to eight serial dilutions of each sample. Concentration-effect models were used to identify differentially expressed genes (DEGs) and to calculate effect concentrations (ECs) of DEGs, which could be ranked to investigate low dose response. Furthermore, molecular pathways disrupted by different samples were evaluated by Gene Ontology (GO) enrichment analysis. The ability of RHT for representing bioactivity utilizing both HepG2 and MCF7 was shown to be comparable to the results of previous in vitro bioassays. Finally, the relative potencies of the mixtures indicated by RHT analysis were consistent with the chemical profiles of the samples. RHT analysis with human cells provides an efficient and cost-effective approach to benchmarking mixture of micropollutants and may offer novel insight into the assessment of mixture toxicity in water.

Enantioselective determination of the chiral pesticide isofenphos-methyl in vegetables, fruits, and soil and its enantioselective degradation in pak choi using HPLC with UV detection.

An enantioselective method for the simultaneous determination of the chiral pesticide isofenphos-methyl in vegetables, fruits, and soil has been established using high-performance liquid chromatography with UV detection. The complete enantioseparation was conducted by reversed-phase liquid chromatography with a cellulose-tris-(4-methylbenzoate) chiral stationary phase (CSP) (Lux Cellulose-3). The effects of different mobile phase compositions, temperatures, and flow rates on enantioseparation were also investigated. The experimental and calculated electronic circular dichroism spectra indicate that the first peak is (S)-(+)-isofenphos-methyl and the second peak is (R)-(-)-isofenphos-methyl. Alumina-A and Florisil solid-phase extraction (SPE) columns were used to clean up for vegetable, fruit, and soil samples. The mean recoveries of the two enantiomers ranged from 83.2 to 110.9 % with intra-day relative standard deviations (RSDs) from 3.2 to 10.8 % and inter-day RSDs from 3.6 to 10 %. Good linearity (≥0.9992) was obtained for the two enantiomers in all matrix-matched calibration curves in the range of 0.25 to 20 mg L(-1). The limit of detection for two enantiomers in six matrices was in the range of 0.008 to 0.011 mg kg(-1), and the limit of quantification was estimated to range from 0.027 to 0.037 mg kg(-1). The results indicated that this method was a convenient and dependable approach for the simultaneous determination of isofenphos-methyl enantiomers in food and environmental samples. The stereoselective degradation of isofenphos-methyl in pak choi has shown that the (R)-(-)-isofenphos-methyl isomer (half-life t 1/2 = 2.2 days) degraded faster than the (S)-(+)-isomer (t 1/2 = 1.9 days). Graphical Abstract The enantioselective determination and enantioselective degradation of the chiral pesticide isofenphos-methyl.

Simultaneous determination of chiral pesticide flufiprole enantiomers in vegetables, fruits, and soil by high-performance liquid chromatography.

A simple and reliable method for the simultaneous determination of chiral pesticide flufiprole enantiomers using high-performance liquid chromatography has been established. The separation and determination were performed using reversed-phase chromatography on a carbamoyl-cellulose-type chiral stationary phase, a Lux Cellulose-2 column. The effects of different mobile phase composition on separation were discussed. The absolute configuration of flufiprole enantiomers was measured through the combination of experimental and predicted ECD spectra. An Alumina-N solid-phase extraction (SPE) column was used in the cleanup of the vegetables, fruits, and soil samples. The method was evaluated by the specificity, matrix effect, linearity, precision, accuracy and stability. The mean recoveries of two enantiomers ranged from 86.8 to 98.9 %, with 1.1-6.4 % intra-day relative standard deviation (RSD) and 1.2 to 5.8 % inter-day RSD. Good linearity (R (2) > 0.998) was obtained for all analyte matrix calibration curves within the range of 0.2-20 mg L(-1). The limit of detection for two enantiomers in the six matrices was 0.007-0.008 mg kg(-1), whereas the limit of quantification of two enantiomers in fruits, vegetables, and soil was 0.021-0.025 mg kg(-1). The results confirmed that this method was convenient and accurate for the simultaneous determination of flufiprole enantiomers in food and environmental samples.

A new lignan from Laggera crispata (Vahl) Hepper & J. R. I. Wood.

A new lignan, 9'-O-angelyllariciresinol (1), and 20 known compounds (2-21) were isolated from the petroleum ether fraction of Laggera crispata (Vahl) Hepper & J. R. I. Wood. Their structures were identified by spectral analysis (NMR, IR, UV, and MS). Activity screening showed that compound 5 exhibited significant inhibitory effect on Staphylococcus aureus, while compound 2 exhibited significant inhibitory effect against liver cancer cell line HepG2.

Impacts of PFOS, PFOA and their alternatives on the gut, intestinal barriers and gut-organ axis.

With the restricted use of perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA), a number of alternatives to PFOS and PFOA have attracted great interest. Most of the alternatives are still characterized by persistence, bioaccumulation, and a variety of toxicity. Due to the production and use of these substances, they can be detected in the atmosphere, soil and water body. They affect human health through several exposure pathways and especially enter the gut by drinking water and eating food, which results in gut toxicity. In this review, we summarized the effects of PFOS, PFOA and 9 alternatives on pathological changes in the gut, the disruption of physical, chemical, biological and immune barriers of the intestine, and the gut-organ axis. This review provides a valuable understanding of the gut toxicity of PFOS, PFOA and their alternatives as well as the human health risks of emerging contaminants.

[Diagnosis of nitrogen content in upper and lower corn leaves based on hyperspectral data].

Based on the spectral characters of corn leaf nitrogen content in the space, the spectral models for rapid estimating crop nitrogen content were set up, which is practically meaningful to effectively providing the guidance in fertilization. Spectral technology was applied to explore corn leaves nitrogen content distribution regularity and the relationship between the nitrogen content and plant index was analysed and then the estimation models were built. The results showed N content in upper leaves is higher than that in lower leaves in four growing stages; lower leaves at tassel emerge stage are sensitive to nitrogen losses, which could be used in guiding fertilization in grain production; optimum estimation models were built atjointing stage, the full-grown stage and tasseling stage, The research results provided the proof of crop nutrient analysis and rational fertilization.

Human functional genomics reveals toxicological mechanism underlying genotoxicants-induced inflammatory responses under low doses exposure.

Understanding the toxicological mechanisms of chemicals is essential for accurate assessments of environmental health risks. Inflammation could play a critical role in the adverse health outcomes caused by genotoxicants; however, the toxicological mechanisms underlying genotoxicants-induced inflammatory response are still limited. Here, functional genomics CRISPR screens were performed to enhance the mechanistic understanding of the genotoxicants-induced inflammatory response at low doses exposure. Key genes and pathways associated with the activities of immune cells and the production of cytokines were identified by CRISPR screens of 6 model genotoxicants. Gene network analysis revealed that three genes (TLR10, HCAR2 and TRIM6) were involved in the regulation of neutrophil apoptosis and cytokine release, and TLR10 shared a similar functional pattern with HCAR2 and TRIM6. Furthermore, adverse outcome pathway (AOP) network analysis revealed that TLR10 was involved in the molecular initiating events (MIEs) or key events (KEs) in the inflammatory response AOPs of all the 6 genotoxicants, which provided mechanistic links between TLR10 and genotoxicants-induced inflammation and respiratory diseases. Finally, functional validation tests demonstrated that TLR10 exhibited inhibitory effects on genotoxicants-induced inflammatory responses in both epithelial and immune cells. This study highlights the powerful utility of the integration of CRISPR screen and AOP network analysis in illuminating the toxicological causal mechanisms of environmental chemicals.

Gut Microbiota and Immune Checkpoint Inhibitors-Based Immunotherapy.

Application of immune checkpoint inhibitors (ICIs) is a major breakthrough in the field of cancer therapy, which has displayed tremendous potential in various types of malignancies. However, their response rates range widely in different cancer types and a significant number of patients experience immune-related adverse effects (irAEs) induced by these drugs, limiting the proportion of patients who can truly benefit from ICIs. Gut microbiota has gained increasing attention due to its emerging role in regulating the immune system. In recent years, numerous studies have shown that gut microbiota can modulate antitumor response, as well as decrease the risk of colitis due to ICIs in patients receiving immunotherapy. The present review analyzed recent progress of relevant basic and clinical studies in this area and explored new perspectives to enhance the efficacy of ICIs and alleviate associated irAEs via manipulation of the gut microbiota.

Assessment of genotoxic chemicals using chemogenomic profiling based on gene-knockout library in Saccharomyces cerevisiae.

Understanding the adverse effects of genotoxic chemicals and identifying them effectively from non-genotoxic chemicals are of great worldwide concerns. Here, Saccharomyces cerevisiae (yeast) genome-wide single-gene knockout screening approach was conducted to assess two genotoxic chemicals (4-nitroquinoline-1-oxide (4-NQO) and formaldehyde (FA)) and environmental pollutant dichloroacetic acid (DCA, genotoxicity is controversial). DNA repair was significant enriched in the gene ontology (GO) biology process (BP) terms and KEGG pathways when exposed to low concentrations of 4-NQO and FA. Higher concentrations of 4-NQO and FA influenced some RNA metabolic and biosynthesis pathways. Moreover, replication and repair associated pathways were top ranked KEGG pathways with high fold-change for low concentrations of 4-NQO and FA. The similar gene profiles perturbed by DCA with three test concentrations identified, the common GO BP terms associated with aromatic amino acid family biosynthetic process and ubiquitin-dependent protein catabolic process via the multivesicular body sorting pathway. DCA has no obvious genotoxicity as there was no enriched DNA damage and repair pathways and fold-change of replication and repair KEGG pathways were very low. Five genes (RAD18, RAD59, MUS81, MMS4, and BEM4) could serve as candidate genes for genotoxic chemicals. Overall, the yeast functional genomic profiling showed great performance for assessing the signatures and potential molecular mechanisms of genotoxic chemicals.

Bis[tris-(ethyl-enediamine-κN,N')cobalt(III)] octa-kis-µ-(3)-oxido-hexa-deca-µ(2)-oxido-tetra-deca-oxido-µ(12)-tetra-oxo-silicato-octa-molybdenum(VI)hexa-vanadium(IV,V) hexa-hydrate.

The title compound, [Co(C(2)H(8)N(2))(3)](2)[SiMo(8)V(4)O(40)(VO)(2)]·6H(2)O, was prepared under hydro-thermal conditions. The asymmetric unit consists of a transition metal complex [Co(en)(3)](3+) cation (en is ethyl-enediamine), one half of an [SiMo(8)V(4)O(40)(VO)(2)](6-) heteropolyanion, two solvent water mol-ecules in general positions and two half-mol-ecules of water located on a mirror plane. In the complex cation, the Co(3+) ion is in a distorted octa-hedral coordination environment formed by six N atoms of the three chelating en ligands. One of the en ligands exhibits disorder of its aliphatic chain over two sets of sites of equal occupancy. The [SiMo(8)V(4)O(40)(VO)(2)](6-) heteropolyanion is a four-electron reduced bivanadyl-capped α-Keggin-type molybdenum-vanadium-oxide cluster. In the crystal, it is located on a mirror plane, which results in disorder of the central tetra-hedral SiO(4) group: the O atoms of this group occupy two sets of sites related by a mirror plane. Furthermore, all of the eight µ(2)-oxide groups are also disordered over two sets of sites with equal occupancy. There are extensive inter-molecular N-H⋯O hydrogen bonds between the complex cations and inorganic polyoxidoanions, leading to a three-dimensional supra-molecular network.

µ(3)-Dodeca-tungsto(V,VI)aluminato-κO:O':O''-tris-[aqua-bis-(ethyl-ene-diamine-κN,N')copper(II)].

The title compound, [AlCu(3)W(12)O(40)(C(2)H(8)N(2))(6)(H(2)O)(3)], was prepared under hydro-thermal conditions. The Cu(2+) ion displays an elongated octa-hedral geometry defined by one bridging O atom from the polyoxidoanion and a coordinated water mol-ecule in axial positions and four N atoms of the two chelating ethyl-enediamine (en) ligands in equatorial positions. The one-electron reduced [AlW(12)O(40)](6-) anion coordinates three [Cu(en)(H(2)O)](2+) fragments, generating a neutral tri-supported Keggin-type polyoxidometalate (POM). This tri-supported POM is located in a special position of [Formula: see text] symmetry and therefore O atoms from the central AlO(4) tetra-hedron are disordered over two sets of sites. Disorder is also observed for three other bridging O atoms of the POM. In the crystal, mol-ecules are connected via N-H⋯O and O-H⋯O hydrogen bonds, forming a three-dimensional framework.

Molecular fingerprints of conazoles via functional genomic profiling of Saccharomyces cerevisiae.

Conazoles were designed to inhibit ergosterol biosynthesis. Conazoles have been widely used as agricultural fungicides and are frequently detected in the environment. Although conazoles have been reported to have adverse effects, such as potential carcinogenic effects, the underlying molecular mechanisms of toxicity remain unclear. Here, the molecular fingerprints of five conazoles (propiconazole (Pro), penconazole (Pen), tebuconazole (Teb), flusilazole (Flu) and epoxiconazole (Epo)) were assessed in Saccharomyces cerevisiae (yeast) via functional genome-wide knockout mutant profiling. A total of 169 (4.49%), 176 (4.67%), 198 (5.26%), 218 (5.79%) and 173 (4.59%) responsive genes were identified at three concentrations (IC50, IC20 and IC10) of Pro, Pen, Teb, Flu and Epo, respectively. The five conazoles tended to have similar gene mutant fingerprints and toxicity mechanisms. "Ribosome" (sce03010) and "cytoplasmic translation" (GO: 0002181) were the common KEGG pathway and GO biological process term by gene set enrichment analysis of the responsive genes, which suggested that conazoles influenced protein synthesis. Conazoles also affected fatty acids synthesis because "biosynthesis of unsaturated fatty acids" pathway was among the top-ranked KEGG pathways. Moreover, two genes, YGR037C (acyl-CoA-binding protein) and YCR034W (fatty acid elongase), were key fingerprints of conazoles because they played vital roles in conazole-induced toxicity. Overall, the fingerprints derived from the yeast functional genomic screening provide an alternative approach to elucidate the molecular mechanisms of environmental pollutant conazoles.

Mechanistic Insights into Stereospecific Bioactivity and Dissipation of Chiral Fungicide Triticonazole in Agricultural Management.

Research interest in chiral pesticides has increased probably because enantiomers often exhibit different environmental fate and toxicity. An investigation into the enantiomer-specific bioactivity of chiral triticonazole enantiomers in agricultural systems revealed intriguing experimental and theoretical evidence. For nine of the phytopathogens studied ( Rhizoctonia solani, Fusarium verticillioide, Botrytis cinerea (strawberry and tomato), Rhizoctonia cereali, Alternaria solani, Gibberella zeae, Sclerotinia sclerotiorum, and Pyricularia grisea), the fungicidal activity data showed ( R)-triticonazole was 3.11-82.89 times more potent than the ( S) enantiomer. Furthermore, ( R)-triticonazole inhibited ergosterol biosynthesis and cell membrane synthesis 1.80-7.34 times higher than its antipode. Homology modeling and molecular docking studies suggested the distinct bioactivities of the enantiomers of triticonazole were probably due to their different binding modes and affinities to CYP51b. However, field studies demonstrated that ( S)-triticonazole was more persistent than ( R)-triticonazole in fruits and vegetables. The results showed that application of pure ( R)-triticonazole, with its high bioactivity and relatively low resistance risk, instead of the racemate in agricultural management would reduce the application dosage required to eliminate carcinogenic mycotoxins and any environmental risks associated with this fungicide, yielding benefits in food safety and environmental protection.

Hepatocellular carcinoma suppressor 1 promoter hypermethylation in serum. A diagnostic and prognostic study in hepatitis B.

BACKGROUND: Liver cancer ranks as the second leading cause of cancer-related mortality in man worldwide, and hepatocellular carcinoma (HCC) is the most prevalent malignant neoplasm of the liver. The sensitivity of alpha-fetoprotein (AFP) as an HCC diagnostic marker for HCC diagnosis is 39-65%, and one-third patients with HCC are missed using AFP. New biomarkers are needed to diagnose HCC at an earlier stage and to individualize treatment strategies. Hepatocellular carcinoma suppressor 1 (HCCS1) is a newly identified liver tumor suppressor gene. OBJECTIVE: Our study evaluated the diagnostic value of serum HCCS1 promoter methylation in patients with HCC associated with hepatitis B. METHODS: We determined the methylation status of serum HCCS1 promoter in 120 patients with HCC, 146 patients with chronic hepatitis B (CHB) and 27 healthy controls (HCs) by methylation-specific polymerase chain reaction (MSP). Evaluation of a cohort with 63 patients with HCC and 44 patients with CHB was set as a validation dataset. RESULTS: The frequency of HCCS1 promoter methylation in patients with HCC was significantly higher than that in patients with CHB (P<0.001) and HCs (P<0.001), and was associated with tumor node-metastasis (TNM) stage (P=0.01). The sensitivity of serum HCCS1 promoter methylation for discriminating patients with HCC from CHB was 62.5% and that of AFP alone was 55%. Notably, the sensitivity of serum HCCS1 promoter methylation plus AFP level was 81.7%. CONCLUSION: HCCS1 has potential as a biomarker for diagnosis and prognosis of patients with HCC.

[Tripartite motif-containing protein 34 (TRIM34) colocalized with micronuclei chromosome and hampers its movement to equatorial plate during the metaphase stage of mitosis].

Objective To examine whether tripartite motif-containing protein 34 (TRIM34) is colocalized with micronuclei and investigate the influence on the movement of micronuclei chromosome in mitosis. Methods The eukaryotic expression vector TRIM34-pEGFP-N3 was constructed, identified and then transfected into HEK293T cells. With 4', 6-diamidino-2-phenylindole 2HCI (DAPI) staining, the colocalization between TRIM34 and micronuclei was observed under a fluorescence microscope. Moreover, MitoTracker(R)Deep Red was used to identify the colocalization between the complex of TRIM34-micronulei and mitochondria under a confocal microscope. Finally, the effect of TRIM34 on the movement of micronuclei chromosome in mitosis was examined. Results DNA sequencing confirmed that the vector TRIM34-pEGFP-N3 was constructed successfully. A fluorescence microscope revealed that TRIM34 could be colocalized with micronuclei in HEK293T cells transfected with TRIM34-pEGFP-N3. In the same manner, a confocal microscope distinctly showed that TRIM34 was colocalized with micronuclei similarly in appearance. However, there was no distinguished colocalization relationship between the complex of TRIM34-micronulei and mitochondria. Interestingly, the micronuclei chromosome conjugated with TRIM34 was hardly transferred to equatorial plate during the metaphase stage of mitosis. Conclusion TRIM34 is colocalized with micronuclei chromosome and hampers its movement to equatorial plate in mitosis.