Epigenetic marks or not? The discovery of novel DNA modifications in eukaryotes.

DNA modifications add another layer of complexity to the eukaryotic genome to regulate gene expression, playing critical roles as epigenetic marks. In eukaryotes, the study of DNA epigenetic modifications has been confined to 5mC and its derivatives for decades. However, rapid developing approaches have witnessed the expansion of DNA modification reservoirs during the past several years, including the identification of 6mA, 5gmC, 4mC, and 4acC in diverse organisms. However, whether these DNA modifications function as epigenetic marks requires careful consideration. In this review, we try to present a panorama of all the DNA epigenetic modifications in eukaryotes, emphasizing recent breakthroughs in the identification of novel DNA modifications. The characterization of their roles in transcriptional regulation as potential epigenetic marks is summarized. More importantly, the pathways for generating or eliminating these DNA modifications, as well as the proteins involved are comprehensively dissected. Furthermore, we briefly discuss the potential challenges and perspectives, which should be taken into account while investigating novel DNA modifications.

A vitamin-C-derived DNA modification catalysed by an algal TET homologue.

Methylation of cytosine to 5-methylcytosine (5mC) is a prevalent DNA modification found in many organisms. Sequential oxidation of 5mC by ten-eleven translocation (TET) dioxygenases results in a cascade of additional epigenetic marks and promotes demethylation of DNA in mammals1,2. However, the enzymatic activity and function of TET homologues in other eukaryotes remains largely unexplored. Here we show that the green alga Chlamydomonas reinhardtii contains a 5mC-modifying enzyme (CMD1) that is a TET homologue and catalyses the conjugation of a glyceryl moiety to the methyl group of 5mC through a carbon-carbon bond, resulting in two stereoisomeric nucleobase products. The catalytic activity of CMD1 requires Fe(II) and the integrity of its binding motif His-X-Asp, which is conserved in Fe-dependent dioxygenases3. However, unlike previously described TET enzymes, which use 2-oxoglutarate as a co-substrate4, CMD1 uses L-ascorbic acid (vitamin C) as an essential co-substrate. Vitamin C donates the glyceryl moiety to 5mC with concurrent formation of glyoxylic acid and CO2. The vitamin-C-derived DNA modification is present in the genome of wild-type C. reinhardtii but at a substantially lower level in a CMD1 mutant strain. The fitness of CMD1 mutant cells during exposure to high light levels is reduced. LHCSR3, a gene that is critical for the protection of C. reinhardtii from photo-oxidative damage under high light conditions, is hypermethylated and downregulated in CMD1 mutant cells compared to wild-type cells, causing a reduced capacity for photoprotective non-photochemical quenching. Our study thus identifies a eukaryotic DNA base modification that is catalysed by a divergent TET homologue and unexpectedly derived from vitamin C, and describes its role as a potential epigenetic mark that may counteract DNA methylation in the regulation of photosynthesis.

Cosinusoidal encoding multiplexed structured illumination multispectral ghost imaging.

The information dimension obtained by multispectral ghost imaging is more abundant than in single-band ghost imaging. Existing multispectral ghost imaging systems still meet some shortages, such as complex structure or reconstruction time-consuming. Here, an approach of cosinusoidal encoding multiplexed structured illumination multispectral ghost imaging is proposed. It can capture the multispectral image of the target object within one projection cycle with a single-pixel detector while maintaining high imaging efficiency and low time-consuming. The core of the proposed approach is the employed novel encoding strategy which is apt to decode and reconstruct the multispectral image via the Fourier transform. Specifically, cosinusoidal encoding matrices with specific frequency characteristics are fused with the orthogonal Hadamard basis patterns to form the multiplexed structured illumination patterns. A broadband photomultiplier is employed to collect the backscattered signals of the target object interacted by the corresponding structured illumination. The conventional linear algorithm is applied first to recover the mixed grayscale image of the imaging scene. Given the specific frequency distribution of the constructed cosinusoidal encoding matrices, the mixed grayscale image can be converted to the frequency domain for further decoding processing. Then, the pictures of multiple spectral components can be obtained with some manipulations by applying Fourier transform. A series of numerical simulations and experiments verified our proposed approach. The present cosinusoidal encoding multiplexed structured illumination can also be introduced in many other fields of high-dimensional information acquisition, such as high-resolution imaging and polarization ghost imaging.

PTEN-silencing combined with ChABC-overexpression in adipose-derived stem cells promotes functional recovery of spinal cord injury in rats.

The efficiency of cell therapy after spinal cord injury (SCI) depend on the survival of transplanted cells. However, sterile microenvironment and glial scar hyperplasia extremely reduce their numbers. Our previous study found overexpression of ChABC gene is positively correlated to migration ability. Expression of PTEN gene is closely associated with proliferation. However, whether manipulation of PTEN and ChABC on adipose-derived mesenchymal stem cells (ADSCs) promote motor recovery is unknown. This study aimed to promote hindlimb function recovery in SCI rats by enhancing proliferation and migration ability of ADSCs, transiently silencing expression of PTEN following overexpression of ChABC (double-gene modified ADSCs, DG-ADSCs). After PTEN silencing, we observed strong proliferation and accelerated G1-S transition in DG-ADSCs using CCK8 assay and flow cytometry. In addition, we demonstrated that migration numbers of DG-ADSCs were higher than control group using Transwell assay. The protein and mRNA levels of MAP2 and ßⅢ-tubulin in DG-ADSCs were increased compared with ADSCs. These results were further confirmed in SCI rats. Increased survival cells and reduction of glial scars were quantitatively analyzed in DG-ADSCs groups, which is definitely correlated to function recovery. Recovery of motor function was observed in DG-ADSCs treatment rats using BBB score, which emphasized that improved viability of transplanted cells and reduction of glial scars were an effective strategy for enhancing recovery of neurological function after SCI.

[Chemical profiling and tissue distribution study of Jingyin Granules in rats using UHPLC-Q-Exactive Orbitrap HR-MS].

In this study, the chemical profiling of Jingyin Granules and the tissue distribution of nine major constituents in this Chinese medicine were performed after oral administration of Jingyin Granules to rats, by using UHPLC-Q-Exactive Orbitrap HR-MS. An Acquity UPLC BEH C_(18) chromatographic column(2.1 mm×100 mm, 1.7 µm) was used as solid phase, while the mobile phase was methanol and 0.1% formic acid water for gradient elution. The major constituents in this Chinese medicine were quickly and accurately identified, via comparison with the retention times and MS/MS spectra of the standards. A total of 106 chemicals were identified from Jingyin Granules, including 24 kinds of organic acids, 47 kinds of flavonoids, 10 kinds of iridoids, and 21 kinds of saponins and 4 kinds of other compounds. After oral administered Jingyin Granules to rats, 48, 30, 25, 23, 45, 34, 39, 26, 19 prototype compounds were identified in serum, heart, liver, spleen, lung, kidney, brain, fat, and testicles, respectively. Meanwhile, an LC-MS based analytical method was established for simultaneous determination of chlorogenic acid, swertiamarin, caffeic acid, sweroside, liquiritin, prim-O-glucosylcimifugin, arctiin, 5-O-methylvisammioside and arctigenin in biological samples. The tissue distribution(serum, liver and lung) of these nine aim constituents in rats after oral administration of Jingyin Granules were investigated. It was found that these nine constituents could be quickly absorbed into circulation system and then distributed to liver and lung tissues. Except arctigenin, the exposure of other eight aim constituents to serum and lung was peaked at 1 h. At 1 h, the exposure of these components to lung tissue were ranked as follows: swertiamarin [(75 191.0±3 483.21) ng·g~(-1)]>arctiin [(2 716.5±36.06) ng·g~(-1)]>5-O-methylvisammioside [(585.1±0.71) ng·g~(-1)]>arctigenin [(437.45±3.18) ng·g~(-1)]>chlorogenic acid [(308.1±5.66) ng·g~(-1)]>prim-O-glucosylcimifugin [(211.35±2.19) ng·g~(-1)]>sweroside [(184.3±9.05) ng·g~(-1)]>caffeic acid [(175.95±2.05) ng·g~(-1)]>liquiritin [(174.78±153.34) ng·g~(-1)]. In summary, an UHPLC-Q-Exactive Orbitrap HR-MS method has been established for rapid and accurate identification of the constituents in Jingyin Granules, while the tissue distribution of nine major absorpted constituents were investigated in rats following oral administration of Jingyin Granules. These findings provided key information and guidance for further studies on pharmacodynamic substances and clinical applications of Jingyin Granules.

Sparse Fourier single-pixel imaging.

Fourier single-pixel imaging is one of the main single-pixel imaging techniques. To improve the imaging efficiency, some of the recent method typically select the low-frequency and discard the high-frequency information to reduce the number of acquired samples. However, sampling only a small amount of low-frequency components will lead to the loss of object details and will reduce the imaging resolution. At the same time, the ringing effect of the restored image due to frequency truncation is significant. In this paper, a new sparse Fourier single-pixel imaging method is proposed that reduces the number of samples explorations while maintaining increased image quality. The proposed method makes a special use of the characteristics of the Fourier spectrum distribution based on which the power of image information decreases gradually from low to high frequencies in the Fourier space. A variable density random sampling matrix is employed to achieve random sampling with Fourier single-pixel imaging technology, followed by the processing of the sparse Fourier spectra using compressive sensing algorithms to recover the high-quality information of the object. The new algorithm can effectively improve the quality of object restoration comparing with the existing Fourier single-pixel imaging methods which only acquire the low-frequency parts. Additionally, considering that the resolution of the system is diffraction limited, super-resolution imaging can also be achieved. Experimental results demonstrate the mainly correctness but also effectiveness of the proposed method.

Radon single-pixel imaging with projective sampling.

A novel technique for Radon single-pixel imaging with projective sampling, which is based on the theorem of the Radon transform, is proposed. In contrast to current patterns in conventional single-pixel imaging systems, candy-striped patterns called Radon basis patterns, which are produced by projecting the 1D Hadamard functions along different angles, are employed in our proposed technique. Here, the patterns are loaded into a projection system and then illuminated onto an object. The light reflected from the object is detected by a single-pixel detector. An iterative reconstruction method is used to restore the object's 1D projection functions by summing the 1D Hadamard functions and detected intensities. Next, the Radon spectrum of the object is recovered by arranging the 1D projection functions along the projection angle. Finally, the image of the object can be recovered using a filtered back-projection algorithm with the Radon spectrum. Experiments demonstrate that the proposed technique can obtain the information of the Radon spectrum and image of the object. Recognition directly in the Radon spectrum domain, rather than in the image domain, is fast and yields robust and high classification rates. A recognition experiment is performed by detecting the lines in one scene by searching the singular peaks in the Radon spectrum domain. According to the results, the lines in the scene can be easily detected in the Radon spectrum domain. Other shapes can also be detected by the characteristics of those shapes in the Radon spectrum domain.

Quantum Spin Liquid Intertwining Nematic and Superconducting Order in Fese.

Despite its seemingly simple composition and structure, the pairing mechanism of FeSe remains an open problem due to several striking phenomena. Among them are nematic order without magnetic order, nodeless gap and unusual inelastic neutron spectra with a broad continuum, and gap anisotropy consistent with orbital selection of unknown origin. Here we propose a microscopic description of a nematic quantum spin liquid that reproduces key features of neutron spectra. We then study how the spin fluctuations of the local moments lead to pairing within a spin-fermion model. We find the resulting superconducting order parameter to be nodeless s±d wave within each domain. Further we show that orbital dependent Kondo-like coupling can readily capture observed gap anisotropy. Our prediction calls for inelastic neutron scattering in a detwinned sample.

Uracil-DNA Glycosylase UNG Promotes Tet-mediated DNA Demethylation.

In mammals, active DNA demethylation involves oxidation of 5-methylcytosine (5mC) into 5-formylcytosine (5fC) and 5-carboxylcytosine (5caC) by Tet dioxygenases and excision of these two oxidized bases by thymine DNA glycosylase (TDG). Although TDG is essential for active demethylation in embryonic stem cells and induced pluripotent stem cells, it is hardly expressed in mouse zygotes and dispensable in pronuclear DNA demethylation. To search for other factors that might contribute to demethylation in mammalian cells, we performed a functional genomics screen based on a methylated luciferase reporter assay. UNG2, one of the glycosylases known to excise uracil residues from DNA, was found to reduce DNA methylation, thus activating transcription of a methylation-silenced reporter gene when co-transfected with Tet2 into HEK293T cells. Interestingly, UNG2 could decrease 5caC from the genomic DNA and a reporter plasmid in transfected cells, like TDG. Furthermore, deficiency in Ung partially impaired DNA demethylation in mouse zygotes. Our results suggest that UNG might be involved in Tet-mediated DNA demethylation.

Endoplasmic Reticulum Stress Promotes Autophagy and Apoptosis and Reduces Chemotherapy Resistance in Mutant p53 Lung Cancer Cells.

BACKGROUND/AIMS: Lung cancer (LC) continues to be one of the most prevalent cancers around the world. During this study we aimed to investigate the involvement of endoplasmic reticulum stress (ERS) in autophagy, apoptosis, and chemotherapy resistance of mutant p53 LC cells. METHODS: Immunohistochemistry was employed to help determine the p53 mutation status of cancer cells from 92 primary LC patients, who were subsequently assigned to either the mutant p53 (n = 39) or wild-type p53 group (n = 53). RESULTS: Mutant p53 cells exhibited increased expression of the C/EBP homologous protein (CHOP), glucose-regulated protein 78 (GRP78), and inositol-requiring enzyme-1α (IRE1α). The Mutant p53 cells were also found to be sensitive to chemotherapy and displayed decreased expression of PI3K, Akt, and mTOR. The mutant p53 cell lines were treated with tunicamycin to induce ERS and rapamycin in order to inhibit mTOR. Both agents increased the expression of CHOP, GRP78, IRE1α, LC3-II/LC3-I, Atg5, Atg7, caspase-3, caspase-12, cleaved caspase-3, cleaved caspase-12, as well as decreases in cell proliferation as well as the expression levels of PI3K, Akt, and mTOR. Enhanced levels of cell apoptosis and reduced chemotherapy resistance were also detected. CONCLUSION: The findings of our study suggest that ERS promotes autophagy and apoptosis, while acting to reduce chemotherapy resistance in mutant p53 LC cells by downregulating the PI3K/Akt/mTOR signaling pathway.

Inhibition of microRNA-196a might reverse cisplatin resistance of A549/DDP non-small-cell lung cancer cell line.

We aimed to explore the possible mechanism of microRNA-196a (miR-196a) inhibition and reversion of drug resistance to cisplatin (DDP) of the A549/DDP non-small-cell lung cancer (NSCLC) cell line. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect expression differences of miR-196a in the drug-resistant A549/DDP NLCLC cell line and the parental A549 cell line, and expressions of miR-196a in the A549/DDP NLCLC cell line transfected with miR-196a inhibitor (anti-miR-196a group) and the miR-196a negative control (miR-NC) group and blank group (without transfection). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test was applied in examining the cell viability of A549/DDP cell line before and after transfection. Clonogenic assay was used to detect cell proliferation ability. Flow cytometry was applied in detecting apoptosis rate of assayed tumor cell and rhodamine-123 changes in cells. Western blot was applied in detecting proteins of drug-resistant related gene in A549/DDP cell line. Significantly higher expression of miR-196a was detected in the drug-resistant A549/DDP cell line than that in the parental A549 cell line (P < 0.05). However, miR-196a expression in the anti-miR-196a group decreased obviously compared to that in the blank group and the miR-NC group (both P < 0.05); The value of IC50 in the anti-miR-196a group showed remarkably lower than that in the blank group and the miR-NC group (both P < 0.05); Rh-123 absorbing ability in the anti-miR-196a group increased 2.51 times and 2.49 times respectively compared to that in the blank group and the miR-NC group (both P < 0.05). No statistical differences in the apoptosis rate of A549/DDP cell line in the early stage were found among the three groups (all P > 0.05), but the late-stage apoptosis rate in the anti-miR-196a group was significantly higher than that in the blank group and the miR-NC group (both P < 0.05); The expressions of human multidrug resistance 1 (MDR1), multidrug resistance protein 1 (MRP1), excision repair cross-complementation 1 (ERCC1), survivin, and B cell lymphoma 2 (Bcl-2) decreased significantly while RhoE increased significantly in the anti-miR-196a group than the blank group and the miR-NC group (all P < 0.05). Inhibition of miR-196a could reverse cisplatin resistance of A549/DDP cell lines, which might relate with inhibition of drug efflux, down-regulation of drug-resistant protein expression, cell apoptosis, and cell proliferation suppression.

The vascular delta-like ligand-4 (DLL4)-Notch4 signaling correlates with angiogenesis in primary glioblastoma: an immunohistochemical study.

Delta-like ligand-4 (DLL4)-Notch signaling is known to play a pivotal role in the regulation of tumor angiogenesis. We had previously found that DLL4 was overexpressed, while Notch1 receptor, which binds to DLL4 during angiogenesis, was absent in the majority of human primary glioblastomas. Thus, DLL4-Notch signaling pathway in the regulation of tumor angiogenesis in primary glioblastoma remains unknown. Tumor tissues from 70 patients with primary glioblastoma were analyzed by immunohistochemistry for expression of components of DLL4-Notch signaling, vascular endothelial growth factor (VEGF), and microvessel density (MVD). Immunohistochemistry results showed that the positive staining of DLL4 and Notch4 was primarily distributed in tumor vascular endothelial cells but rarely detected in tumor cells. However, VEGF, hairy/enhancer of split-1 (HES1; a target gene of Notch signaling), and Notch1-3 expression was seen in both tumor vascular endothelial cells and tumor cells. Univariate analysis showed that the expression levels of VEGF and DLL4, HES1, and Notch4 in tumor endothelial cells were significantly associated with MVD in primary glioblastoma (P < 0.001). Binary logistic regression analysis showed that high expression levels of DLL4, HES1, and Notch4 in tumor endothelial cells were associated with a decrease of MVD in primary glioblastoma, while MVD increased with elevated VEGF expression in contrast. In addition, DLL4, Notch4, and HES1 expression were positively correlated in tumor vascular endothelial cells (P < 0.05). We conclude that the vascular DLL4-Notch4 signaling and VEGF signaling complementing each other plays an important role in the progression of tumor angiogenesis in primary glioblastoma. Graphical abstract A, positive staining of DLL4 in human kidney; B, positive staining of VEGF in human breast cancer; C, positive staining of CD34 in human lung cancer; D, positive staining of HES1 in human breast cancer; E-H, positive staining of Notch1-4: E-F in human lung cancer; G-H in human kidney.

One-stage posterior focus debridement, interbody grafts, and posterior instrumentation and fusion in the surgical treatment of thoracolumbar spinal tuberculosis with kyphosis in children: a preliminary report.

PURPOSE: The purpose of this study was to determine the efficacy and feasibility of surgical management of children with thoracolumbar spine tuberculosis with kyphosis by using one-stage posterior focus debridement, interbody grafts, and posterior instrumentation and fusion. METHODS: From October 2010 to September 2013, 21 children with thoracolumbar spinal tuberculosis accompanied by kyphosis were treated with one-stage posterior decompression, interbody grafts, and posterior instrumentation and fusion. There were 13 males and 8 females, aged from 7 to 13 years old (average age 9.9 years). The mean follow-up was 34 months (range26-48 months). Patients were evaluated before and after surgery in terms of ESR, neurologic status, pain, and kyphotic angle. RESULTS: Spinal tuberculosis was completely cured, and the grafted bones were fused in all 21 patients. There was no recurrent tuberculous infection. ESR got normal within 3 months in all patients. The ASIA neurologic classification improved in all cases. Pain relief was obtained in all patients. The average preoperative kyphosis was 29.7° (range 12-42°) and decreased to 5.5° (range 2-10°), postoperatively. There was no significant loss of the correction at the latest follow-up. CONCLUSIONS: Our results show that one-stage posterior decompression, interbody grafts, and posterior instrumentation and fusion were an effective treatment for children with thoracolumbar spinal tuberculosis. It is characterized as minimum surgical trauma, good neurologic recovery, good correction of kyphosis, and prevention of progressive kyphosis.

Effects of a feeding intervention in patients with Alzheimer's disease and dysphagia.

AIMS AND OBJECTIVES: This study aimed to investigate the effects of a feeding intervention in patients with Alzheimer's disease with dysphagia. BACKGROUND: In patients with Alzheimer's disease, inadequate food and fluid intake can result in malnutrition, dehydration and increased morbidity and mortality. Patients may lose self-care abilities such as self-feeding. DESIGN: A prospective cohort study. METHODS: A three-month self-control feeding intervention was conducted prospectively in 30 nursing home residents with Alzheimer's disease with dysphagia. Pre- and post-intervention measures included the Kubota water swallow test, type and amount of food intake and assessment of nutritional status by triceps skinfold thickness, upper arm circumference, serum albumin and haemoglobin. We used the Edinburgh Feeding Evaluation in Dementia scale to evaluate eating compliance and the Mini- Mental State Examination to evaluate cognitive function. Pre- and post-intervention results were compared to evaluate the effects of nursing intervention. RESULTS: Patients' eating/feeding abilities improved overall, including significantly increased food intake (p < 0·001), decreased levels on the Kubota water swallow test (p < 0·001) and significant differences in skinfold thickness, arm circumference, serum albumin and haemoglobin (all p < 0·01), indicating improved nutritional status. Edinburgh Feeding Evaluation in Dementia scale scores decreased significantly, showing improved eating compliance. No changes were noted in cognition post-intervention. Among 22 patients who initially required feeding, five patients resumed self-feeding after the intervention (p = 0·06). CONCLUSIONS: Results of this study show that a feeding intervention can improve food intake, eating compliance and nutritional status in patients with Alzheimer's disease with dysphagia and prevent further decline in swallowing function. RELEVANCE TO CLINICAL PRACTICE: The significant improvement in eating/feeding measures suggest that this feeding intervention model could be developed as a feeding skills programme to improve both the eating/feeding care by nursing staff and the eating/feeding abilities and nutritional status of Alzheimer's disease patients.

One-stage posterior focus debridement, interbody graft using titanium mesh cages, posterior instrumentation and fusion in the surgical treatment of lumbo-sacral spinal tuberculosis in the aged.

PURPOSE: Aged patients represent a high risk group for acquiring spinal tuberculosis, and it still remains a leading cause of kyphosis and paraplegia in developing nations. Aged patients often combined with cardiovascular and respiratory disease and single lung ventilation via anterior approach surgery could result in more post-operative complications. We aimed to analyze the efficacy and feasibility of surgical management of aged patients with lumbo-sacral spine tuberculosis using one-stage posterior focus debridement, interbody graft using titanium mesh cages, posterior instrumentation, and fusion. METHODS: From March 2009 and July 2012, 17 aged patients with lumbo-sacral spinal tuberculosis were treated with one-stage posterior focus debridement, interbody graft using titanium mesh cages, posterior instrumentation, and fusion. There were eight male and nine female with a mean age of 63.3 years (range: 60-71 years). The mean follow-up was 46.5 months (range 38-70 months). Patients were evaluated before and after surgery in terms of ESR, neurological status, visual analog scale (VAS), and lumbosacral angle. RESULTS: Spinal tuberculosis was completely cured and the grafted bones were fused in all 17 patients. There were no recurrent tuberculous infections. ESR became normal within three months in all patients. The ASIA neurological classification and VAS scores improved in all cases. The average preoperative lumbosacral angle was 20.6° (range 18.1°-22.5°) and became 29.4° (range 27.1°-32.5°) at final follow-up. CONCLUSIONS: Our results showed that one-stage posterior focus debridement, interbody graft using titanium mesh cages, posterior instrumentation, and fusion was an effective treatment for aged patients with lumbo-sacral spinal tuberculosis. It is characterized with minimum surgical trauma, good pain relief, good neurological recovery, and good reconstruction of the spinal stability.

Effectiveness of adaptive optics system in satellite-to-ground coherent optical communication.

Adaptive optics (AO) systems can suppress the signal fade induced by atmospheric turbulence in satellite-to-ground coherent optical communication. The lower bound of the signal fade under AO compensation was investigated by analyzing the pattern of aberration modes for a one-stage imaging AO system. The distribution of the root mean square of the residual aberration is discussed on the basis of the spatial and temporal characteristics of the residual aberration of the AO system. The effectiveness of the AO system for improving the performance of coherent optical communication is presented in terms of the bit error rate and system availability.

Identification of candidate biomarkers for early detection of human lung squamous cell cancer by quantitative proteomics.

To discover novel biomarkers for early detection of human lung squamous cell cancer (LSCC) and explore possible mechanisms of LSCC carcinogenesis, iTRAQ-tagging combined with two dimensional liquid chromatography tandem MS analysis was used to identify differentially expressed proteins in human bronchial epithelial carcinogenic process using laser capture microdissection-purified normal bronchial epithelium (NBE), squamous metaplasia (SM), atypical hyperplasia (AH), carcinoma in situ (CIS) and invasive LSCC. As a result, 102 differentially expressed proteins were identified, and three differential proteins (GSTP1, HSPB1 and CKB) showing progressively expressional changes in the carcinogenic process were selectively validated by Western blotting. Immunohistochemistry was performed to detect the expression of the three proteins in an independent set of paraffin-embedded archival specimens including various stage tissues of bronchial epithelial carcinogenesis, and their ability for early detection of LSCC was evaluated by receiver operating characteristic analysis. The results showed that the combination of the three proteins could perfectly discriminate NBE from preneoplastic lesions (SM, AH and CIS) from invasive LSCC, achieving a sensitivity of 96% and a specificity of 92% in discriminating NBE from preneoplatic lesions, a sensitivity of 100% and a specificity of 98% in discriminating NBE from invasive LSCC, and a sensitivity of 92% and a specificity of 91% in discriminating preneoplastic lesions from invasive LSCC, respectively. Furthermore, we knocked down GSTP1 in immortalized human bronchial epithelial cell line 16HBE cells, and then measured their susceptibility to carcinogen benzo(a)pyrene-induced cell transformation. The results showed that GSTP1 knockdown significantly increased the efficiency of benzo(a)pyrene-induced 16HBE cell transformation. The present data first time show that GSTP1, HSPB1 and CKB are novel potential biomarkers for early detection of LSCC, and GSTP1 down-regulation is involved in human bronchial epithelial carcinogenesis.

[Establishment of composite reference intervals for thyroid-stimulating hormone and free thyroxine in determination of subclinical hypothyroidism].

OBJECTIVE: To detect changes of thyroid stimulating hormone (TSH) with gender, age and levels of thyroid peroxidase antibodies (TPO-Ab) in patients with subclinical hypothyroidism, and to establish composite reference intervals for TSH and free thyroxine (FT4) in determination of subclinical hypothyroidism. METHODS: From Oct. 2011 to July 2012, 7 964 healthy people (males: 4 789, females: 3 175) undergoing medical examinations were recruited. Their serum levels of TSH and FT4 were determined. Of those participants, 794 were also tested for TPO-Ab. The serum TSH and FT4 data were transformed into normal distributions, with outliers being eliminated and a correction for skewness (0. 909 and 0. 384, respectively) and kurtosis (2.605 and 1.947, respectively). The composite reference intervals were established according to the Mahalonobis distance formula. RESULTS: Serum TSH increased with age and TPO-Ab. Using the conventional reference standards, 358 participants were identified with subclinical hypothyroidism, which included 230 at 41-70 years of age and 43 showing TPO-Ab positive. In contrast, using composite reference intervals, 301 participants were identified with subclinical hypothyroidism, which included 142 at 41-70 years of age and 25 showing TPO-Ab. CONCLUSION: The conventional cutoff values for FT4 and TSH separately lead to overestimation of the prevalence of subclinical thyroid disease.

Bioinformatics analysis revealed the potential crosstalk genes and molecular mechanisms between intracranial aneurysms and periodontitis.

OBJECTIVES: The risk of intracranial aneurysms (IAs) development and rupture is significantly higher in patients with periodontitis (PD), suggesting an association between the two. However, the specific mechanisms of association between these two diseases have not been fully investigated. MATERIALS AND METHODS: In this study, we downloaded IAs and PD data from the Gene Expression Omnibus. Differentially expressed genes (DEGs) were identified, and functional enrichment analysis was performed. The protein-protein interaction (PPI) network and weighted gene co-expression network analysis (WGCNA) was performed to identified key modules and key crosstalk genes. In addition, the immune cell landscape was assessed and the correlation of key crosstalk genes with each immune cell was calculated. Finally, transcription factors (TFs) regulating key crosstalk genes were explored. RESULTS: 127 overlapping DEGs were identified and functional enrichment analysis highlighted the important role of immune reflection in the pathogenesis of IAs and PD. We identified ITGAX and COL4A2 as key crosstalk genes. In addition, the expression of multiple immune cells was significantly elevated in PDs and IAs compared to controls, and both key crosstalk genes were significantly negatively associated with Macrophages M2. Finally, GATA2 was identified as a potential key transcription factor (TF), which regulates two key crosstalk gene. CONCLUSIONS: The present study identifies key crosstalk genes and TF in PD and IAs, providing new insights for further study of the co-pathogenesis of PD and IAs from an immune and inflammatory perspective. Also, this is the first study to report the above findings.

The potential crosstalk genes and molecular mechanisms between glioblastoma and periodontitis.

Despite clinical and epidemiological evidence suggestive of a link between glioblastoma (GBM) and periodontitis (PD), the shared mechanisms of gene regulation remain elusive. In this study, we identify differentially expressed genes (DEGs) that overlap between the GEO datasets GSE4290 [GBM] and GSE10334 [PD]. Functional enrichment analysis was conducted, and key modules were identified using protein-protein interaction (PPI) network and weighted gene co-expression network analysis (WGCNA). The expression levels of CXCR4, LY96, and C3 were found to be significantly elevated in both the test dataset and external validation dataset, making them key crosstalk genes. Additionally, immune cell landscape analysis revealed elevated expression levels of multiple immune cells in GBM and PD compared to controls, with the key crosstalk genes negatively associated with Macrophages M2. FLI1 was identified as a potential key transcription factor (TF) regulating the three key crosstalk genes, with increased expression in the full dataset. These findings contribute to our understanding of the immune and inflammatory aspects of the comorbidity mechanism between GBM and PD.