Long non-coding RNAs (lncRNAs) are essential factors that regulate tumor development and metastasis via diverse molecular mechanisms in a broad type of cancers. However, the pathological roles of ...lncRNAs in gallbladder carcinoma (GBC) remain largely unknown. Here we discovered a novel lncRNA termed lncRNA Highly expressed in GBC (lncRNA-HGBC) which was upregulated in GBC tissue and aimed to investigate its role and regulatory mechanism in the development and progression of GBC.
The expression level of lncRNA-HGBC in GBC tissue and different cell lines was determined by quantitative real-time PCR. The full length of lncRNA-HGBC was obtained by 5' and 3' rapid amplification of the cDNA ends (RACE). Cellular localization of lncRNA-HGBC was detected by fluorescence in situ hybridization (FISH) assays and subcellular fractionation assay. In vitro and in vivo assays were preformed to explore the biological effects of lncRNA-HGBC in GBC cells. RNA pull-down assay, mass spectrometry, and RNA immunoprecipitation (RIP) assay were used to identify lncRNA-HGBC-interacting proteins. Dual luciferase reporter assays, AGO2-RIP, and MS2-RIP assays were performed to verify the interaction between lncRNA-HGBC and miR-502-3p.
We found that lncRNA-HGBC was upregulated in GBC and its upregulation could predict poor survival. Overexpression or knockdown of lncRNA-HGBC in GBC cell lines resulted in increased or decreased, respectively, cell proliferation and invasion in vitro and in xenografted tumors. LncRNA-HGBC specifically bound to RNA binding protein Hu Antigen R (HuR) that in turn stabilized lncRNA-HGBC. LncRNA-HGBC functioned as a competitive endogenous RNA to bind to miR-502-3p that inhibits target gene SET. Overexpression, knockdown or mutation of lncRNA-HGBC altered the inhibitory effects of miR-502-3p on SET expression and downstream activation of AKT. Clinically, lncRNA-HGBC expression was negatively correlated with miR-502-3p, but positively correlated with SET and HuR in GBC tissue.
Our study demonstrates that lncRNA-HGBC promotes GBC metastasis via activation of the miR-502-3p-SET-AKT cascade, pointing to lncRNA-HGBC as a new prognostic predictor and a therapeutic target.
LiNi1/3Co1/3Mn1/3O2 as a promising cathode material in lithium‐ion batteries was synthesized by flash/field‐assisted sintering technique for the first time. This study showed that the current‐limited ...synthesis of LiNi1/3Co1/3Mn1/3O2 could be carried out at temperatures less than 400°C for only 8 minutes, compared with the conventional pressureless sintering at 850°C for 12 hours. X‐ray diffraction results showed the phase evolution from precursor mixtures to the final LiNi1/3Co1/3Mn1/3O2 products during flash/field‐assisted sintering process and a well‐layered structure without undesirable cation mixing in the as‐formed LiNi1/3Co1/3Mn1/3O2. Combined with the lowered sintering temperatures and reduced sintering time, the excellent electrochemical performance of flash/field‐assisted sintered LiNi1/3Co1/3Mn1/3O2 materials suggested that this technique could be an energy‐efficient approach for the synthesis of lithium‐ion battery cathode materials and other materials requiring high‐temperature heat treatment.
Abstract
Background
To evaluate the value of the real‐time PCR–based multicolor melting curve analysis (MMCA) with an automatic analysis system used in a mass thalassemia screening and prenatal ...diagnosis program.
Methods
A total of 18,912 peripheral blood samples from 9456 couples and 1150 prenatal samples were detected by MMCA assay. All prenatal samples were also tested by a conventional method. Samples with unknown melting peaks, unusual peak height ratios between a wild allele and a mutant allele, or a discordant phenotype‐genotype match were further studied by using multiplex ligation–dependent probe amplification (MLPA) or Sanger sequencing. All MMCA results were automatically analyzed and manually checked. The consistency between MMCA assay and conventional methods among prenatal samples was investigated.
Results
Except for initiation codon (T > G) (HBB:c.2T > G), all genotypes of thalassemia inside the scope of conventional methods were detected by MMCA assay. Additionally, 27 carriers with 10 rare HBB variants, 13 with α fusion gene, 1 with a rare deletion in α globin gene, and 1 with rare HBA variant were detected by using MMCA assay.
Conclusion
MMCA can be an alternative approach used in routine thalassemia carrier screening and prenatal diagnosis for its high throughput, sufficient stability, low cost, and easy operation.
Gallbladder cancer (GBC) is the most common malignant tumour of the biliary track system. Angiogenesis plays a pivotal role in the development and progression of malignant tumours. miR-143-3p acts as ...a tumour suppressor in various cancers. Their role in GBC is however less well defined. Here we show that the expression levels of miR-143-3p were decreased in human GBC tissues compared with the non-tumour adjacent tissue (NAT) counterparts and were closely associated with overall survival. We discovered that miR-143-3p was a novel inhibitor of tumour growth and angiogenesis in vivo and in vitro. Our antibody array, ELISA and PLGF rescue analyses indicated that PLGF played an essential role in the antiangiogenic effect of miR-143-3p. Furthermore, we used miRNA target-prediction software and dual-luciferase assays to confirm that integrin α6 (ITGA6) acted as a direct target of miR-143-3p. Our ELISA and western blot analyses confirmed that the expression of PLGF was decreased via the ITGA6/PI3K/AKT pathway. In conclusion, miR-143-3p suppresses tumour angiogenesis and growth of GBC through the ITGA6/PI3K/AKT/PLGF pathways and may be a novel molecular therapeutic target for GBC.
To explore mutations in the additional sex combs-like 3 (
ASXL3
) gene in two Chinese families with congenital heart disease (CHD). Whole-exome sequencing (WES) was used to reveal a novel compound ...heterozygous mutation in the
ASXL3
gene that was associated with CHD. Sanger sequencing of a further 122 CHD patients was used to determine an additional compound heterozygous mutation in the
ASXL3
gene. Cell apoptosis was examined by MTS assay and flow cytometry. The cardiac structure was identified via hematoxylin–eosin (HE), Masson’s trichrome, and ultrasound scanning. RNA sequencing was performed to identify a series of differentially expressed mRNAs. The mRNA and protein expressions were identified by quantitative real-time PCR and western blotting, respectively. A compound heterozygous mutation c.2168C > G (p.Pro723Arg) and c.5449C > G (p.Pro1817Ala) in the
ASXL3
gene associated with CHD was identified. Overexpression of this compound heterozygous mutation in HL-1 cells resulted in increased apoptosis and reduced cell viability. Moreover, it affected cardiac structure and fibrosis in mice. There were 126 downregulated mRNAs and 117 upregulated mRNAs between the
ASXL3
compound heterozygous mutation c.2168C > G (p.Pro723Arg) and c.5449C > G (p.Pro1817Ala) mice and wild-type mice.
Ezh2
,
Slc6a4
, and
Socs3
, which could interact with
ASXL3
through proteins, were all upregulated. Another compound heterozygous mutation c.3526C > T (p.Arg1176Trp) and c.4643A > G (p.Asp1548Gly) in the
ASXL3
gene was identified by screening a further 122 patients with CHD. The
ASXL3
gene is important in cardiac development and may exert this influence by affecting the expression of mRNAs associated with cell apoptosis and cell proliferation.
Our study was aimed to explore the clinical implication of chromosome microarray analysis (CMA) in genetically etiological diagnosis of children with congenital heart disease (CHD).
A total of 104 ...children with CHD with or without multiple congenital anomalies (MCA) or intellectual disabilities/developmental delay (ID/DD) but normal karyotype were investigated using Affymetrix CytoScan HD array.
Pathogenic copy number variations (PCNVs) were identified in 29 children (27.9%). The detection rates in children with simple CHD and complex CHD were 31.1% (19/61) and 23.2% (10/43), respectively. The detection rates of PCNVs were 17.9% (7/39), 20% (5/25), 63.2% (12/19) and 23.8% (5/21) in isolated CHD, CHD plus MCA, CHD plus ID/DD, CHD plus MCA and ID/DD, respectively. The PCNVs rate of CHD plus ID/DD was significantly higher than that of isolated CHD. Two genomic loci including 15q11.2 deletion and 1q43-q44 deletion were considered as CHD locus. The DVL1, SKI, STIM1, CTNNA3 and PLN were identified as candidate genes associated with CHD phenotypes.
CMA can increase the diagnostic rate and improve the etiological diagnosis in children with CHD. We suggest CMA as a first-tier test in children with CHD, especially in children with CHD plus ID/DD.
•Chemical composition of waters in upper Huanghe is mainly controlled by carbonate weathering.•Evaporite dissolution in the midstream is responsible for the high TDS in Huanghe basin.•Waters ...composition in the lower reaches nearly inherits the feature of middles reaches.•Carbonate weathering in upper reaches is the main sink of CO2 in the Huanghe basin.
To systematically study chemical weathering in the entire Huanghe River basin, we divided the basin into three parts with significant differences in geology and climate. We collected 38 water samples from the main channel and its main tributaries, spread between the upstream (above Lanzhou), midstream (from Lanzhou to Huayuankou) and downstream (from Huayuankou up to river mouth) segments of the river. The concentrations of major elements and H, O isotopic compositions were obtained from the samples, and the total dissolved solids (TDS) and, CO2 consumption budget were calculated from the aggregated data of each of the three segments. The results demonstrate that: the TDS are mainly derived from carbonate weathering in upper Huanghe River; evaporite dissolution is predominately occurred in the midstream; and there is almost no additional contribution from rock weathering in the downstream. An increasing trend of CO2 consumption rate by silicate weathering is observed, from 0.14×105mol/km2/yr in the upstream to 5.62×105mol/km2/yr in the downstream, and the budget of CO2 consumption by silicate weathering is estimated to be 26.2×109mol/yr. In contrast, the CO2 consumption rate by carbonate weathering decreases from 3.04×105mol/km2/yr in the upstream to near zero in the downstream, and the budget of CO2 consumption is estimated to be 100.5×109mol/yr. As a whole, in the entire Huanghe River basin, the CO2 consumption budget and TDS yield are estimated to be 126.7×109mol/yr, and 27.5×106t/yr, respectively. These results indicate that evaporite dissolution in the midstream is responsible for the high TDS in the Huanghe River basin, while carbonate weathering in the upstream plays the most significant role in CO2 consumption.
Gallbladder carcinoma (GBC), the most common malignant tumour of the bile duct, is highly aggressive and has a poor prognosis. MicroRNA-30a-5p (miR-30a-5p) is an important tumour suppressor that ...participates in many aspects of carcinogenesis and cancer development. However, the role of miR-30a-5p in GBC development remains to be determined, as do the mechanisms underlying its effects in GBC. Using samples collected from 42 subjects with gallbladder carcinoma (GBC), we showed decreased miR-30a-5p expression in the primary lesions vs. non-tumour adjacent tissues (NATs). Decreased miR-30a-5p was associated with shorter disease-free survival (DFS) and overall survival (OS). Inhibiting miR-30a-5p expression in 2 representative GBC cell lines (GBC-SD and NOZ) increased cell proliferation, migration, invasiveness, as well as β-catenin nuclear translocation, vice versa. In nude mice, NOZ cells transfected with miR-30a-5p mimics grew slower (vs. miR-NC) upon subcutaneous inoculation, and had lower rate of hepatic metastasis upon spleen inoculation. Dual luciferase assay confirmed that E2F transcription factor 7 (E2F7) was a direct target of miR-30a-5p and antagonized the effects induced by miR-30a-5p downregulation in GBC cells. MiR-30a-5p attenuates the EMT and metastasis in GBC cells by targeting E2F7, suggesting miR-30a-5p is a tumour suppressor that may serve as a novel potential prognostic biomarker or molecular therapeutic target for GBC.
Xinqiao is a large copper-gold deposit and consists of two major mineralization types: stratabound and skarn. The skarn occurs along the contact between a quartz diorite intrusion and ...Carboniferous-Triassic limestone. Xinqiao has a strongly developed skarn zone, including endoskarn and exoskarn; the exoskarn is divided into proximal and distal exoskarn. We present systematic major, trace and rare earth element (REE) concentrations for garnets from the skarn zone, discuss the factors controlling the incorporation of trace elements into the garnets, and constrain the formation and evolution of the garnet from skarn zone in Xinqiao deposit. Grossular (Adr20–44Grs56–80) mostly occurs in endoskarn and has typical HREE-enriched and LREE-depleted patterns, with small Eu anomalies and low ∑REE. Garnets from the exoskarn show complex textures and chemical compositions. The composition of garnets range from Al-rich andradite (Adr63–81Grs19–47) to andradite (Adr67–98Grs2–33). Garnet in endoskarn has typical HREE-enriched and LREE-depleted patterns. Al-rich andradite in proximal skarn has small Eu anomalies and moderate ∑REE. Andradite from distal exoskarn shows strong positive Eu anomalies and has variable ∑REE. The U, Y, Fe and Al relationship with ∑REE shows that two mechanisms controlled incorporation of REE into the garnets: crystal chemistry (substitution and interstitial solid solution) mainly controlled in the endoskarn garnet (grossular) and the proximal exoskarn (Al-rich andradite), and fluid and rock chemistry (surface adsorption and occlusion) controlled REEs in the distal exoskarn. Furthermore, Al has a negative relationship with ∑REE indicating that REE3+ did not follow a coupled, YAG-type substitution into the garnets. Variations in textures and trace and rare earth elements of garnets suggest that the garnets in the endoskarn formed by slow crystal growth at low W/R ratios and near-neutral pH in a closed system during periods of diffusive metasomatism. The garnets in the exoskarn formed rapidly from externally derived fluids during advective metasomatism, and adsorption had a major control on the REE patterns in distal exoskarn. With the end of water-rock reaction, the contents of REE decreased in the hydrothermal fluid, and the system became nearly closed.
•Xinqiao has a strongly developed skarn zone, including endoskarn and exoskarn.•Garnets from endoskarn to exoskarn have a distinct ∑REE and δ Eu variation.•The variation of ∑REE and δ Eu in garnet can indicate the formation and evolution of garnet in skarn zone.
A hybrid variable neighborhood search (HVNS) algorithm, which combines the key characteristics of chemical-reaction optimization (CRO) and estimation of distribution (EDA), for solving the hybrid ...flow shop (HFS) scheduling problems. The proposed algorithm is tested on sets of the well-known benchmark instances. Through the analysis of experimental results, the highly effective performance of the proposed HVNS algorithm is shown against the best performing algorithms from the literature.
•We propose a novel hybrid variable neighborhood search (HVNS) algorithm for hybrid flow shop (HFS) scheduling problems.•A well-designed decoding mechanism is presented to schedule jobs with more flexibility.•Considering the problem structure, eight neighborhood structures are developed.•A kinetic energy sensitive neighborhood change approach is proposed.•A dynamic neighborhood set update mechanism is utilized.•An effective EDA-based global search approach is presented.
This paper proposes a hybrid variable neighborhood search (HVNS) algorithm that combines the chemical-reaction optimization (CRO) and the estimation of distribution (EDA), for solving the hybrid flow shop (HFS) scheduling problems. The objective is to minimize the maximum completion time. In the proposed algorithm, a well-designed decoding mechanism is presented to schedule jobs with more flexibility. Meanwhile, considering the problem structure, eight neighborhood structures are developed. A kinetic energy sensitive neighborhood change approach is proposed to extract global information and avoid being stuck at the local optima. In addition, contrary to the fixed neighborhood set in traditional VNS, a dynamic neighborhood set update mechanism is utilized to exploit the potential search space. Finally, for the population of local optima solutions, an effective EDA-based global search approach is investigated to direct the search process to promising regions. The proposed algorithm is tested on sets of well-known benchmark instances. Through the analysis of experimental results, the high performance of the proposed HVNS algorithm is shown in comparison with four efficient algorithms from the literature.
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