Significance of screening sensitive methylation sites using whole-genome sequencing in early diagnosis of non-small cell lung cancer.

The current study aimed to screen the sensitive methylation sites of non-small cell lung cancer by whole-genome sequencing and construct an early warning system for lung cancer. For this purpose, from June 2017 to December 2020, fresh NSCLC tissues and paired adjacent NSCLC tissues from 45 patients were collected. DNA and total RNA were extracted from non-small cell lung cancer (NSCLC) and paired non-cancerous lung tissues. The DNA library combined with a biotinylated probe was collected by Dynabeads m270 streptavidin beads. The concentration of the final library was determined by qubit dsDNA HS assay. Quantitative analysis of DMR methylation in 45 paired tumor and normal lung tissues was performed. RT qPCR and Western blot were used to verify the mRNA expression of candidate genes. Results showed that the methylation rate of CpG 7 in stxbp6 in stage III NSCLC was higher than that in stage I and early-stage II NSCLC; The methylation rates of cpg1 and 38-39 units in fzd10 were higher in stage I NSCLC than in stage II and III NSCLC; The methylation rates of CpG 6 in stxbp6 and CpG 4 and 20-21 in bcl6b in patients with tumor diameter > 3cm were higher than those in patients with tumor diameter < 3cm; Methylation of CpG unit 3 in stxbp6 is associated with age. Stxbp6, bcl6b, fzd10 and hspb6 mRNA expression were down-regulated in patients under 45 years old. The methylation rates of CpG 7 in stxbp6, CPG 6 in stxbp6 and CpG 4 and 20-21 in bcl6b were negatively correlated with the survival time of patients; The methylation rates of CpG 1 and 38-39 units in fzd10 were positively correlated with survival time (P<0.05). It was concluded that the methylation rates of CpG 7 in stxbp6, CPG 6 in stxbp6 and CpG 4 and 20-21 in bcl6b are valuable for early diagnosis.

In vitro and in vivo toxic effects and inflammatory responses induced by carboxylated black carbon-lead complex exposure.

Black carbon (BC) is a key component of atmospheric fine particulate matter (PM2.5) and it tends to adsorb various pollutants (e.g., heavy metals and organics) during atmospheric transport. This adsorption leads to the complexity and uncertainty of the source and chemical composition of PM2.5, making the toxicologic effects and health risks induced by PM2.5 difficult to determine. Here, we used carboxylated black carbon (c-BC) and c-BC-lead complexes (c-BC-Pb) to investigate the in vitro and in vivo toxic effects and inflammatory responses. The physicochemical properties of c-BC and c-BC-Pb complexes were characterized by the transmission electron microscope (TEM), X-ray photoelectron spectroscopy (XPS), dynamic light scattering (DLS), and in ductively coupled plasma-atomic emission spectra (ICP-AES). Cytotoxicity in vitro showed that the exposure of human bronchial epithelial cells (BEAS-2B) to low-dose c-BC-Pb particles significantly induced greater toxicity than that of c-BC, suggesting that lead (Pb) might play an important role in induced cytotoxicity after combined exposure to c-BC-Pb particles. The findings were further confirmed by the results in vivo, which indicated that c-BC-Pb particles significantly induced inflammation and lung injury. Based on the results of this experiment, the differences in toxicity can be attributed to the synergistic effect of Pb on the BC particles, which play a synergistic role in vitro and in vivo in the development of toxicity. The c-BC-Pb particles model used in this study may be helpful for the evaluation of cytotoxicity induced by different sources of BC particles or BC-heavy metal complexes and provide a new approach for understanding PM2.5-induced toxicity and health risks.

Decreased SIRT1 protein may promote HMGB1 translocation in the keratinocytes of patients with cutaneous lupus erythematosus.

Background Ultraviolet radiation causes DNA damage in keratinocytes, aggravating cutaneous lupus erythematosus (CLE). High mobility group box 1 (HMGB1) participates in nucleotide excision and may transfer from the nucleus to the cytoplasm in immune active cells and the translocation of HMGB1 may result in DNA repair defects. HMGB1 was observed to transfer from the nucleus to the cytoplasm in the keratinocytes of CLE patients. As a class III histone deacetylases (HDACs), sirtuin-1 (SIRT1) can induce HMGB1 deacetylation. Epigenetic modification of HMGB1 may lead to HMGB1 translocation. Aims We aimed to evaluate the expressions of SIRT1 and HMGB1 in the epidermis of CLE patients and whether decreased SIRT1 leads to HMGB1 translocation through HMGB1 acetylation in keratinocytes. Methods We measured the messenger RNA (mRNA) and protein expressions of SIRT1 and HMGB1 in CLE patients using real-time reverse transcription polymerase chain reaction (RT-qPCR) and western blotting. Keratinocytes were treated with SIRT1 activator resveratrol (Res) and irradiated with ultraviolet B (UVB). We detected the localization expression of HMGB1 by immunofluorescence. The apoptosis level and the cell cycle proportions were measured by flow cytometry. The acetyl-HMGB1 level was detected by immunoprecipitation. Results Compared to healthy controls, the mRNA and protein expressions of SIRT1 in the epidermis of CLE patients were significantly decreased and there was translocation of HMGB1 from the nucleus to the cytoplasm. In keratinocytes, UVB irradiation led to HMGB1 translocation from the nucleus to the cytoplasm. Res treatment inhibited HMGB1 translocation, attenuated the cell apoptosis induced by UVB and decreased the acetyl-HMGB1 level. Limitations We only treated keratinocytes with the SIRT1 activator but did not perform the relevant experiments in keratinocytes with SIRT1 knockdown or overexpression. In addition, the lysine residue site of action of SIRT1 deacetylation of HMGB1 is unclear. The specific mechanism of action of SIRT1 deacetylation of HMGB1 needs to be further investigated. Conclusion SIRT1 may inhibit HMGB1 translocation by HMGB1 deacetylation which inhibited the apoptosis of keratinocytes induced by UVB. Decreased SIRT1 may promote HMGB1 translocation in the keratinocytes of patients with CLE.

[Geographical variation of growth traits among different Quercus acutissima provenances]. / ä¸åŒç§æºéº»æ Žç”Ÿé•¿æ€§çŠ¶çš„åœ°ç†å˜å¼‚.

The variations in plant growth of 32 Quercus acutissima provenances in three sites (Yongfeng, Jiangxi; Kaihua, Zhejiang and Chuzhou, Anhui) were studied. The AMMI model was used to analyze the stability of growth traits and to select the best provenances. The results showed that tree height, diameter at breast (ground) height, and aboveground biomass (AGB) per individual of the provenances were significantly different in the three sites. The AGB per individual was significantly affected by site, provenance, and the interaction of provenance × site. Site had the greatest impact on the variation of tree growth, followed by provenance and provenance × site. The growth performance of these superior provenances in different sites varied greatly in the seedling stage (1-3 years old) and young forest stage (4-11 years old). Based on the AGB per individual in the 11st year, the best provenances were selected in each site. Seven superior provenances of Yongfeng, Jiangxi were selected, with the average AGB per individual increased by 15.6%-57.8% compared with the ave-rage value. Seven superior provenances of Kaihua, Zhejiang were selected, with the average AGB per individual increased by 19.2%-45.2%. Eight superior provenances of Chuzhou, Anhui were selected,with the average AGB per individual increased by 24.9%-63.3%. According to the growth performance and stability, four superior provenances were selected to develop short-rotation charcoal forest cultivation across three sites, with an average AGB per individual of 36.55 kg and an average stability parameter of 0.97.

Insights into the binding behavior of bovine serum albumin to black carbon nanoparticles and induced cytotoxicity.

Black carbon (BC) is a main component of particulate matter (PM2.5). Due to their small size (<100nm), inhaled ultrafine BC nanoparticles may penetrate the lung alveoli, where they interact with surfactant proteins and lipids, causing more serious damage to human health. Here, BC was analyzed to investigate the binding mechanism of its interaction with protein and induction of cytotoxicity changes. The binding process and protein conformation between BC and a serum protein (bovine serum albumin, BSA) were monitored by using a fluorescence quenching technique and UV-vis absorption, Fourier transform infrared (FTIR) and circular dichroism (CD) spectroscopies. The experimental results revealed that the fluorescence quenching of BSA induced by BC was a static quenching process and the hydrophobic force played the critical role in the interaction. The native conformation of BSA on the BC surface was slightly disturbed but obvious structural unfolding of the secondary structure did not occur. In the cytotoxicity study, BC nanoparticles with low concentrations exhibited strong toxicity towards BEAS-2B cells. However, the toxicity of BC nanoparticles could be mitigated by the presence of BSA. Therefore, proteins in biological fluids likely reduce the toxic effect of BC on human health. These findings delineated the binding mechanism and the toxicity between BC and the BSA-BC system, contributing to the understanding of the biological effects of BC exposure on human health in polluted atmospheres.

[Latitude variation mechanism of leaf traits of Metasequoia glyptostroboides in eastern coastal China]. / 沿海地区水杉叶片性状的纬度变化机制.

We analyzed the rules of Metasequoia glyptostroboides along with latitude, including leaf length, leaf width, leaf perimeter, leaf area, ratio of leaf length to width, specific leaf area (SLA), and leaf dry mass based on eight stands growing at different latitudes in the coastal area of eastern China, as well as their relationships with climatic and soil factors. The results showed that the leaf length, leaf width and leaf perimeter increased with increasing latitude, while the leaf area and SLA firstly increased and then decreased. The mean annual temperature and annual precipitation were the major environmental factors affecting the leaf traits along latitude gradient. With the increase of soil N content, the SLA decreased firstly and then increased, while the leaf mass decreased significantly. With the increase of soil P content, the SLA increased, and the leaf mass decreased significantly.

[Variations in leaf C, N, P stoichiometry of Quercus acutissima provenance forests.] / éº»æ Žç§æºæž—å¶ç‰‡ç¢³ã€æ°®ã€ç£·åŒ–å­¦è®¡é‡ç‰¹å¾çš„å˜å¼‚.

In order to explore the variations in leaf stoichiometry based on the method of forest tree breeding, we determined the leaf carbon (C), nitrogen (N), phosphorus (P) stoichiometry among 29 Quercus acutissima provenances grown at 3 sites. The results indicated that the site (environment) effect was statistically significant on leaf stoichiometry, with 13.2%-66.7% of the total variations accounted for leaf C, N, P, C/N, C/P and N/P, while the provenance effect was insignificant and only accounted for 2.9%-11.0% of total variations for leaf stoichiometry. The leaf N and C/N, N and N/P, P and C/P, P and N/P were significantly correlated, and the common standardized major axis slope was also observed among three sites and two provenance groups. It could be concluded that the leaf stoichiometry of Q. acutissima, was mainly determined by its growing environment, due to the similar C, N and P biochemical pathways at species level. The stable correlation coefficients among sites and provenances implied the coupling ratios of leaf stoichiometry were independent of environment and provenance, which supported the leaf stoichiometric homeostasis.

Vaccination with different doses of Dermatophagoides farinae in a mouse model of allergic asthma.

BACKGROUND: The efficacy of vaccination with different doses of a vaccine has not been clarified. The purpose of this study was to investigate the relationship between the doses of purified house dust mite allergen, and allergic airway inflammation and airway reactivity in a mouse model of allergic asthma. METHODS: C57BL/6 mice were sensitized with Dermatophagoides farinae (Der f) and randomly treated with saline, 0.1, 1, or 2 mg Der f vaccine, respectively. The airway hyperresponsiveness, the numbers of total cells and eosinophils and the levels of interferon-γ and interleukin (IL)-4 in the bronchoalveolar lavage fluid (BALF) from individual mice were measured. The percentages of CD11c CD86 and CD11c MHC II+ dendritic cells were characterized, and the levels of IL-12 in the supernatants of cultured dendritic cells were measured. RESULTS: The airway hyperresponsiveness, the numbers of total cells and eosinophils in the BALF and the percentages of CD11c CD86 and CD11c MHC II+ dendritic cells were associated with immunologic downregulation. The levels of IFN-γ in the BALF and the levels of IL-12 in the supernatants of cultured dendritic cells were associated with immunologic upregulation. It was found that vaccination with the medium dose of Der f showed a stronger inhibitory effect. CONCLUSIONS: There seems to be a dose-dependent response between allergen and both airway inflammation and hyperreactivity in the mouse model of allergic asthma.

[Long-term effects of thinning on carbon storage in Cunninghamia lanceolata plantations].

The stand environment and tree growth could be changed as well as carbon storage be affected by thinning. Thus it is important to conduct the research on changes of carbon stock in plantations after thinning for assessing the dynamics of forest ecosystem carbon pool. The carbon storage and its distribution of various components in 22-year-old Cunninghamia lanceolata plantations were studied with control and different treatments such as moderate and heavy thinning. Moderate (thinning intensity was 35%) and heavy (thinning intensity was 50%) thinning treatments were conducted twice at the age of 7 and 14 years, respectively. The stand of control was thinned 15% in the 14th year. The results showed that the proportion of stem carbon storage increased with the increasing thinning intensity, while the proportion of carbon storage in branches, leaves and roots slightly decreased, which suggested that thinning was beneficial for carbon stocking in stem. However, the carbon storage in arbor layer decreased with the thinning intensity in C. lanceolata plantation under moderate and heavy thinning treatments, accounted for 89.0% and 83.1% of the control, respectively. The arbor carbon storage decreased in followed two years after the first thinning. The carbon storage in arbor layer had a fast recovery rate within eight years after the second thinning, and the increment of carbon storage in arbor layer had no difference with the control for the heavy thinning treatment. The carbon storage in understory vegetation, litter and soil layers also had no significant difference under the different thinning treatments. Generally total ecosystem carbon storage under the control, moderate and heavy thinning treatments reached 169.34, 156.65 and 154.37 t x hm(-2), respectively. There was no significant difference among the three treatments. Therefore, it could be concluded that the carbon storage in C. lanceolata plantation did not reduce after thinning in more than 15 years.

[Photosynthetic characters of Quercus acutissima from different provenances under effects of salt stress].

The photosynthetic characteristics of Quercus acutissima from Feixian, Chuzhou and Wanyuan provenances were investigated under NaCl stress at 5 concentrations. For the three provenances, their leaf chlorophyll a (Chl a) and chlorophyll b (Chl b) contents and net photosynthetic rate (Pn) decreased with increasing salt stress, among which, the maximal decrement of Chl a content was 21.5%-36.1% for Feixian provenance, and 3.5%-19.8% for Wanyuan provenance. The diurnal variation of Pn, in control groups had a single peak, and no midday photosynthetic depression was observed. Under high salt stress, all the provenances presented midday photosynthetic depression. Path analysis indicated that the major factors affecting Pn were vapor pressure deficit (VPD) and leaf temperature (Tl) , while the minor factors were stomatal conductance (Gs) , intercellular CO2 concentration (Ci), and photosynthetically active radiation (PARi). In the control groups, the photosynthetic characteristics mainly depended on the climate factors of provenance location. Based on the variations of leaf chlorophyll contents and Pn among the provenances under different salt stress, Wanyuan provenance had the highest salt tolerance, followed by Chuzhou provenance, and Feixian provenance.