LncRNA CALML3-AS1 modulated by m6A modification induces BTNL9 methylation to drive non-small-cell lung cancer progression.

Non-small cell lung cancer (NSCLC) is a common and lethal malignancy. The carcinogenic roles of lncRNA CALML3 antisense RNA 1 (CALML3-AS1) have been documented. However, the function and potential mechanisms of CALML3-AS1 in the progression of NSCLC need to be further explored. The molecule expression was assessed by qRT-PCR and Western blot. The subcellular localization of CALML3-AS1 was observed by fluorescence in situ hybridization (FISH). The malignant behaviors of NSCLC cells were evaluated by CCK-8, colony formation, EdU, wound healing and transwell assays. In vivo xenograft tumor and liver metastatic models were established. The molecular mechanisms were investigated by RIP, RNA pull-down and ChIP assays. The methylation level was detected by MSP. Herein, we found that CALML3-AS1 was upregulated, while butyrophilin-like 9 (BTNL9) was downregulated in NSCLC. Functionally, CALML3-AS1 depletion repressed NSCLC cell malignant phenotypes, in vivo tumor growth, and liver metastasis. Mechanistically, AlkB homolog 5 (ALKBH5) enhanced CALML3-AS1 stability via N6-methyladenosine (m6A) demethylation, whereas m6A reader YTH domain-containing 2 (YTHDC2) destabilized CALML3-AS1. Moreover, CALML3-AS1 inhibited BTNL9 transcription and expression through the recruitment of Zeste homolog 2 (EZH2). Rescue experiments demonstrated that BTNL9 downregulation counteracted sh-CALML3-AS1-mediated antitumor effects on NSCLC. Taken together, CALML3-AS1 modulated by ALKBH5 and YTHDC2 in an m6A modification dependent manner drives NSCLC progression via epigenetically repressing BTNL9.

m6A methyltransferase METTL3-induced lncRNA SNHG17 promotes lung adenocarcinoma gefitinib resistance by epigenetically repressing LATS2 expression.

Gefitinib has been widely applied for the treatment of lung adenocarcinoma (LUAD). However, the long-term application of gefitinib usually leads to acquired drug resistance in tumour patients, resulting in clinical treatment failure. Small nucleolar host gene 17 (SNHG17) has been shown to play a regulatory role in LUAD progression. Nevertheless, the role of SNHG17 in LUAD gefitinib resistance remains elusive. The expression pattern of SNHG17 was examined in tissues and cell lines of gefitinib-sensitive and gefitinib-resistant LUAD, respectively. Gain- and loss-of-function experiments were employed to assess the biological functions of SNHG17 in cell proliferation and apoptosis, as well as aggressive phenotypes of LUAD cells. MeRIP-qPCR and colorimetric quantificational analysis were performed to detect m6A modifications and contents. Fluorescence in situ hybridisation (FISH) and subcellular fractionation analysis were used to reveal the distribution of SNHG17. RIP and ChIP assays were performed to further validate the SNHG17/EZH2/LATS2 regulatory axis. A xenograft tumour growth assay was conducted to evaluate the role of SNHG17 in LUAD gefitinib resistance in vivo. SNHG17 was upregulated in gefitinib-resistant LUAD tissues and cell lines. Functional assays showed that SNHG17 aggravated the malignant phenotypes of gefitinib-resistant LUAD cells. In addition, METTL3-mediated N6-methyladenosine modification could induce the upregulation of SNHG17by stabilising its RNA transcript. Mechanistically, SNHG17 epigenetically repressed the expression of LATS2 by recruiting EZH2 to the promoter region of LATS2. The regulatory role of the SNHG17/EZH2/LATS2 axis in LUAD gefitinib resistance was further supported in vivo. Collectively, our findings suggested that SNHG17 induced by METTL3 could promote LUAD gefitinib resistance by epigenetically repressing LATS2 expression.

TGF-ß1/SH2B3 axis regulates anoikis resistance and EMT of lung cancer cells by modulating JAK2/STAT3 and SHP2/Grb2 signaling pathways.

The pathogenesis of lung cancer, the most common cancer, is complex and unclear, leading to limited treatment options and poor prognosis. To provide molecular insights into lung cancer development, we investigated the function and underlying mechanism of SH2B3 in the regulation of lung cancer. We indicated SH2B3 was diminished while TGF-ß1 was elevated in lung cancer tissues and cells. Low SH2B3 level was correlated with poor prognosis of lung cancer patients. SH2B3 overexpression suppressed cancer cell anoikis resistance, proliferation, migration, invasion, and EMT, while TGF-ß1 promoted those processes via reducing SH2B3. SH2B3 bound to JAK2 and SHP2 to repress JAK2/STAT3 and SHP2/Grb2/PI3K/AKT signaling pathways, respectively, resulting in reduced cancer cell anoikis resistance, proliferation, migration, invasion, and EMT. Overexpression of SH2B3 suppressed lung cancer growth and metastasis in vivo. In conclusion, SH2B3 restrained the development of anoikis resistance and EMT of lung cancer cells via suppressing JAK2/STAT3 and SHP2/Grb2/PI3K/AKT signaling cascades, leading to decreased cancer cell proliferation, migration, and invasion.

GRP78 facilitates M2 macrophage polarization and tumour progression.

This study investigated the regulation of GRP78 in tumour-associated macrophage polarization in lung cancer. First, our results showed that GRP78 was upregulated in macrophages during M2 polarization and in a conditioned medium derived from lung cancer cells. Next, we found that knocking down GRP78 in macrophages promoted M1 differentiation and suppressed M2 polarization via the Janus kinase/signal transducer and activator of transcription signalling. Moreover, conditioned medium from GRP78- or insulin-like growth factor 1-knockdown macrophages attenuated the survival, proliferation, and migration of lung cancer cells, while conditioned medium from GRP78-overexpressing macrophages had the opposite effects. Additionally, GRP78 knockdown reduced both the secretion of insulin-like growth factor 1 and the phosphorylation of the insulin-like growth factor 1 receptor. Interestingly, insulin-like growth factor 1 neutralization downregulated GRP78 and suppressed GRP78 overexpression-induced M2 polarization. Mechanistically, insulin-like growth factor 1 treatment induced the translocation of GRP78 to the plasma membrane and promoted its association with the insulin-like growth factor 1 receptor. Finally, IGF-1 blockade and knockdown as well as GRP78 knockdown in macrophages inhibited M2 macrophage-induced survival, proliferation, and migration of lung cancer cells both in vitro and in vivo.

LncRNA ST7-AS1, by regulating miR-181b-5p/KPNA4 axis, promotes the malignancy of lung adenocarcinoma.

BACKGROUND: Growing evidence suggests that suppressor of tumorigenicity 7 antisense RNA 1 (ST7-AS1) is an oncogenic long noncoding RNA (lncRNA). However, little is known on its clinical significance, biological functions, or molecular mechanisms in lung adenocarcinoma (LUAD). METHODS: The expression of ST7-AS1 and miR-181b-5p were examined by qRT-PCR. The correlations between ST7-AS1 level and different clinicopathological features were analysed. In vitro, LUAD cells were examined for cell viability, migration and invasion by MTT, wound healing and Transwell assay, respectively. Epithelial-mesenchymal transition (EMT) biomarkers were detected by Western blot. The regulations between ST7-AS1, miR-181b-5p, and KPNA4 were examined by luciferase assay, RNA immunoprecipitation, RNA pulldown. Both gain- and loss-of-function strategies were used to assess the importance of different signalling molecules in malignant phenotypes of LUAD cells. The in vivo effect was analysed using the xenograft and the experimental metastasis mouse models. RESULTS: ST7-AS1 was upregulated in LUAD tissues or cell lines, correlated with tumours of positive lymph node metastasis or higher TNM stages, and associated with shorter overall survival of LUAD patients. ST7-AS1 essentially maintained the viability, migration, invasion, and EMT of LUAD cells. The oncogenic activities of ST7-AS1 were accomplished by sponging miR-181b-5p and releasing the suppression of the latter on KPNA4. In LUAD tissues, ST7-AS1 level positively correlated with that of KPNA4 and negatively with miR-181b-5p level. In vivo, targeting ST7-AS1 significantly inhibited xenograft growth and metastasis. CONCLUSIONS: ST7-AS1, by regulating miR-181b-5p/KPNA4 axis, promotes the malignancy of LUAD cells. Targeting ST7-AS1 and KPNA4 or up-regulating miR-181b-5p, therefore, may benefit the treatment of LUAD.

Co-ligand tuned pyrimidine-2-carboxylate Mn(ii) complexes from a 2D 63 layer to an interpenetrated srs-net.

A 63 2D layer complex [Mn3Cl3(L3)]n·H2O (1) (L = pyrimidine-2-carboxylate) was obtained by assembling 2-cyanopyrimidine and manganese chloride, in which the L ligands were generated in situ. In 1 six-membered Mn rings were constructed from MnII ions and L ligands, which were connected to each other by double chloride anions affording a 2D layer. When the chloride anions in 1 were substituted partly by formate, [Mn4Cl3L4(HCO2)]n (2) was obtained. In 2, the L ligands bridge MnII to give a 1D chain, which was further connected by the double chloride anions and Cl/formate bridges to form a two-fold interpenetrated srs-net. Interestingly, 2 exhibits an obvious SHG response of approximately 0.8 times that of KDP. Furthermore, 2 is an antiferromagnet with a field induced spin flop transition.

[An Analysis of the Spectrums between Different Canopy Structures Based on Hyperion Hyperspectral Data in a Temperate Forest of Northeast China].

Canopy is a major structural layer for vegetation to carry out ecological activities. The differences of light radiative transfer processes in canopies caused by forest canopy structure directly influence remote sensing inversion of forest canopy biochemical composition. Thus an analysis of spectral characteristics between different canopy structures contributes to improve the accuracy of remote sensing inversion of forest canopy biochemical components. Based on a Hyperion hyperspectral image in the north Slope of Changbai Mountain Nature Reserve, through FLAASH (the Fast Line-of-sight Atmospheric Analysis of Spectral Hypercubes) atmospheric correction, different canopy reflectance spectra were extracted, and spectral transforms were carried out using continuum removal method and first derivative method for quantitative analysis of the spectral characteristics. A set of spectral indices were calculated, including NIR (near infrared reflectance), NDVI (normalized difference vegetation index), EVI (Enhanced Vegetation Index), NDNI (normalized difference nitrogen index), SPRI (normalized photochemical reflectance index) * NDVI and SPRI * EVI (vegetation productivity index). Combined with the broad foliar dominance index (BFDI), the relationships between the spectral indices and canopy structure composition were investigated. The characteristics of canopy structure composition impacting its spectral curve and indices were clarified in the temperate forest. The results showed that: (1) there existed significantly different spectral characteristics between different canopy structures: comparing to the spectrum of broad-leaved forest canopies, the red edge moved to the left and their slope decreased, blue edge and yellow edge features were also weakened, near-infrared reflectance decreased, normalized reflectance in visible region risen for the spectrum of conifer forest canopies; (2) the spectrum variation were controlled by BFDL The correlations between BFDI and the spectral indices were significant (P < 0.01). It was suggested the ratio of broad-leaved and conifer in canopy played an important role in variation of spectral indices. The coefficients of determination (R2) of BFDI and NDVI, EVI, SPRI * EVI, SPRI * NDVI and NDNI were 0.90, 0.83, 0.83, 0.81, 0.68 and 0.59 respectively. It was revealed that BFDI could control the variation of the canopy structure, greenness, leaf nitrogen concentration, leaf area index and productivity in temperate coniferous and broad-leaved mixed forests. Our findings were very significant foundation for accurate determination of forest type, quantitative extraction of canopy biochemical components, estimation of regional forest ecosystem productivity and other related researches.

[Estimation of light-use efficiency of China' s mid-subtropical planted coniferous forest based on flux measurements and spectral observations].

The photochemical reflectance index (PRI) calculated from spectral reflectance has universally become a proxy for the light-use efficiency (LUE), which significantly improves the LUE-based estimation of ecosystem gross primary productivity on a large scale through upscaling. In this study, we observed the vegetation spectral reflectance of a planted subtropical coniferous forest from the top of a flux tower at Qianyanzhou Station, one of the ChinaFLUX sites, in September and December 2013, and simultaneously measured CO2 flux and meteorological variables for correlation and regression analysis. Results showed that PRI had a better correlation with LUE (R2 = 0.20, P< 0.001) than that of normalized difference vegetation index (NDVI), i.e., PRI was preferred in LUE retrieval. During the whole observation period, PRI and soil water content (SWC)-based bivariate regression model correlated well with LUE (R2 = 0.29, P < 0.001 and R2 = 0.30, P < 0.01 for daytime and midday observation, respectively), but in autumn the bivariate regression model of PRI and vapor pressure deficit (VPD) had a higher correlation with LUE (R2 = 0.448, P < 0.001) for midday observation, which showed that environmental factors, i.e., SWC and VPD, had a potential in improving the LUE retrieval from PRI, but the choice of appropriate environmental factors depended on season.

Single nucleotide polymorphisms of myostatin gene in Chinese domestic horses.

The myostatin gene (MSTN) is a genetic determinant of skeletal muscle growth. Single nucleotide polymorphisms (SNP) in MSTN are of importance due to their strong associations with horse racing performances. In this study, we screened the SNPs in MSTN gene in 514 horses from 15 Chinese horse breeds. Six SNPs (g.26T>C, g.156T>C, g.587A>G, g.598C>T, g.1485C>T, g.2115A>G) in MSTN gene were detected by sequencing and genotyped using PCR-RFLP method. The g.587A>G and g.598C>T residing in the 5'UTR region were novel SNPs identified by this study. The g.2115A>G which have previously been associated with racing performances were present in Chinese horse breeds, providing valuable genetic information for evaluating the potential racing performances in Chinese domestic breeds. The six SNPs together defined thirteen haplotypes, demonstrating abundant haplotype diversities in Chinese horses. Most of the haplotypes were shared among different breeds with no haplotype restricted to a specific region or a single horse breed. AMOVA analysis indicated that most of the genetic variance was attributable to differences among individuals without any significant contribution by the four geographical groups. This study will provide fundamental and instrumental genetic information for evaluating the potential racing performances of Chinese horse breeds.

Y chromosome diversity and paternal origin of Chinese cattle.

To determine the Y chromosome genetic diversity and paternal origin of Chinese cattle, 369 bulls from 17 Chinese native cattle breeds and 30 bulls from Holstein and four bulls from Burma were analyzed using a recently discovered USP9Y marker that could distinguish between taurine and indicine cattle more efficiently. In total, the taurine Y1, Y2 haplogroup and indicine Y3 haplogroup were detected in 7 (1.9 %), 193 (52.3 %) and 169 (45.8 %) individuals of 17 Chinese native breeds, respectively, although these frequencies varied amongst the Chinese native cattle breeds examined. Y2 dominates in northern China (91.4 %), while Y3 dominates in southern China (81.2 %). Central China is an admixture zone with Y2 predominating overall (72.0 %). Our results demonstrate that Chinese cattle have two paternal origins, one from B. taurus (Y2) and the other from B. indicus (Y3). The Y1 haplogroup may originate from the imported beef cattle breeds in western countries. The geographical distributions of the Y2 and Y3 haplogroup frequencies reveal a pattern of male indicine introgression from south to north China, and male taurine introgression from north to south China.

Clinical significance of SH2B1 adaptor protein expression in non-small cell lung cancer.

UNLABELLED: The SH2B1 adaptor protein is recruited to multiple ligand-activated receptor tyrosine kinases that play important role in the physiologic and pathologic features of many cancers. The purpose of this study was to assess SH2B1 expression and to explore its contribution to the non-small cell lung cancer (NSCLC). METHODS: SH2B1 expression in 114 primary NSCLC tissue specimens was analyzed by immunohistochemistry and correlated with clinicopathological parameters and patients' outcome. Additionally, 15 paired NSCLC background tissues, 5 NSCLC cell lines and a normal HBE cell line were evaluated for SH2B1 expression by RT-PCR and immunoblotting, immunofluorescence being applied for the cell lines. RESULTS: SH2B1 was found to be overexpressed in NSCLC tissues and NSCLC cell lines. More importantly, high SH2B1 expression was significantly associated with tumor grade, tumor size, clinical stage, lymph node metastasis, and recurrence respectively. Survival analysis demonstrated that patients with high SH2B1 expression had both poorer disease- free survival and overall survival than other patients. Multivariate Cox regression analysis revealed that SH2B1 overexpression was an independent prognostic factor for patients with NSCLC. CONCLUSIONS: Our findings suggest that the SH2B1 protein may contribute to the malignant progression of NSCLC and could offer a novel prognostic indicator for patients with NSCLC.

[Characteristics of terrestrial ecosystem primary productivity in East Asia based on remote sensing and process-based model].

Based on the bi-linearly interpolated meteorological reanalysis data from National Centers for Environmental Prediction, USA and by using the leaf area index data derived from the GIMMS NDVI to run the process-based Boreal Ecosystems Productivity Simulator (BEPS) model, this paper simulated and analyzed the spatiotemporal characteristics of the terrestrial ecosystem gross primary productivity (GPP) and net primary productivity (NPP) in East Asia in 2000-2005. Before regional simulating and calculating, the observation GPP data of different terrestrial ecosystem in 15 experimental stations of AsiaFlux network and the inventory measurements of NPP at 1300 sampling sites were applied to validate the BEPS GPP and NPP. The results showed that BEPS could well simulate the changes in GPP and NPP of different terrestrial ecosystems, with the R2 ranging from 0.86 to 0.99 and the root mean square error (RMSE) from 0.2 to 1.2 g C x m(-2) x d(-1). The simulated values by BEPS could explain 78% of the changes in annual NPP, and the RMSE was 118 g C x m(-2) x a(-1). In 2000-2005, the averaged total GPP and total NPP of the terrestrial ecosystems in East Asia were 21.7 and 10.5 Pg C x a(-1), respectively, and the GPP and NPP exhibited similar spatial and temporal variation patterns. During the six years, the total NPP of the terrestrial ecosystems varied from 10.2 to 10.7 Pg C x a(-1), with a coefficient of variation being 2. 2%. High NPP (above 1000 g C x m(-2) x a(-1)) occurred in the southeast island countries, while low NPP (below 30 g C x m(-2) x a(-1)) occurred in the desert area of Northwest China. The spatial patterns of NPP were mainly attributed to the differences in the climatic variables across East Asia. The NPP per capita also varied greatly among different countries, which was the highest (70217 kg C x a(-1)) in Mongolia, far higher than that (1921 kg C x a(-1)) in China, and the lowest (757 kg C x a(-1)) in India.

[Carbon sequestration potential of forest vegetation in Jiangxi and Zhejiang provinces based on national forest inventory].

Based on the sixth national forest inventory (1999-2003) and the investigation data of 1030 forest sampling plots in subtropical China collected from publications, and by using stand growth empirical equation, this paper estimated the carbon sequestration potential of forest vegetation under natural growth in Jiangxi and Zhejiang provinces in 2004-2013. In the meanwhile, the effects of three forest management measures, including interplanting, selective thinning, and fertilization, on the future forest carbon sequestration were explored by using the survey data of 455 sampling plots. In 2004-2013, the mean annual carbon sequestration potential of forest vegetation under natural growth in Jiangxi and Zhejiang could reach 11.37 and 4.34 Tg C a(-1) (1 Tg=10(12) g), respectively. Interplanting could generate the largest carbon sequestration potential of forest vegetation, followed by selective thinning, and fertilization, resulting in an increase in the potential by (6.54 +/- 3.9) Tg C a(-1), (3.81 +/- 2.02) Tg C a(-1), and (2.35 +/- 0.6) Tg C a(-1) in Jiangxi and by (2.64 +/- 1.28) Tg C a(-1), (1.42 +/- 0.69) Tg C a(-1), and (1.15 +/- 0.29) Tg C a(-1) in Zhejiang, respectively.

[Profile distribution and accumulation characteristics of organic carbon and total nitrogen in typical marshes in Sanjiang Plain].

The study on the profile distribution and accumulation characteristics of organic carbon (C) and total nitrogen (N) in two typical marshes in Sanjiang Plain showed that on the sediment profiles of Carex lasiocarpa marsh and Phragmites communis marsh, there existed distinct deposition horizon and illuviation horizon. In deposition horizon (0-60 cm), the average contents of organic C and total N were about 96 and 184 g x kg(-1), and 7.4 and 17.6 g x kg(-1), respectively, while in illuviation horizon, they were at low level. The contents of organic C and total N were exponentially decreased with increasing depth, and had significant correlations with bulk density (P < 0.01). The organic C density was the highest at the depth of 20-40 cm. In the deposition horizon of the two marshes, the stocks of organic C and total N were 1.83 x 10(4) and 1.73 x 10(4) t x km(-2), and 1.45 x 10(3) and 1.67 x 10(3) t x km(-2), respectively, and at the depth of 0-100 cm, they were 2.86 x 10(4) and 2.62 x 10(4) t x km(-2), and 2.18 x 10(3) and 2.49 x 10(3) t x km(-2), respectively. Vegetation type had definite effects on the content and stock of organic C and total N in the marsh profiles.

[Leaf nitrogen and phosphorus stoichiometry across 654 terrestrial plant species in NSTEC].

Stoichiometry of leaf N and P is a characteristic of plant to adapt to environment, and can provide data for process-based modeling at large scale. Leaf nitrogen and phosphorus stoichiometry of NSTEC terrestrial plants were studied based on a NSTEC data set including leaf nitrogen and phosphorus observations for 654 plant species at 168 sites. The results showed that leaf nitrogen and phosphorus stoichiometry in NSTEC exhibited large variations, primarily ranging 2.17-52.61 mg x g(-1) for N, 0.10-10.27 mg x g(-1) for P, 1.7-74.6 for N/P ratio. Geometric means for all plant species were 17.55 mg x g(-1), 1.28 mg x g(-1) and 13.5, respectively. Leaf P of NSTEC (ever across China) was lower than global level, therefore plant growth was more limited by P in China region comparing to global environment. For all functional groups, the difference of leaf N was largest, but that of N/P ratio was smallest, since leaf N and P closely related in most of functioning groups; for leaf N and P of the different phylogenic functional groups, the difference was largest for farthest relative (seed vs fern), least for closest relative (monocotyledon vs. dicotyledon) . There were obvious correlation between leaf N (or P) and latitude (or mean annual temperature, MAT). Leaf N and P significantly increased with latitude increasing (or MAT decreasing), but the relationship between N/P ratio and latitude (or MAT) wasn't significant (p = 0.386 and p = 0.342), and the reason maybe include, leaf N and P had the same tendencies and large variations, and region in this research was smaller than global research.

[Spatial characteristics of soil organic carbon and nitrogen in inner Mongolia].

Studies on the amount and spatial distribution of soil organic carbon/nitrogen in Inner Mongolia and their relationship to main climate factors showed that the content of soil organic carbon and nitrogen was 3.24-43.24 kg.m-3 and 269.56-3085.60 g.m-3, respectively, and the C/N ratio was about 4.46-17.13. The correlation between soil organic carbon/nitrogen and temperature was negative, and R was 0.557 and 0.460, respectively. Soil organic carbon/nitrogen had a weak positive correlation to precipitation, and R was 0.285 and 0.203. Soil organic carbon and nitrogen appeared a reducing trend with increasing temperature and decreasing precipitation from northeast to southwest.