Optimizing the Heat Treatment Method to Improve the Aging Response of Al-Fe-Ni-Sc-Zr Alloys.

This work has studied the co-addition of Sc and Zr elements into the Al-1.75wt%Fe-1.25wt%Ni eutectic alloy. The changes in the microstructure, electrical conductivity, and Vickers hardness of the Al-1.75wt%Fe-1.25wt%Ni-0.2wt%Sc-0.2wt%Zr alloy during heat treatment were studied. The results showed that two-step aging can effectively improve the aging response of the alloy over the single-step aging method. This was ascribed to the minimization of the diffusion difference between Sc and Zr elements. Furthermore, the homogenization treatment can also improve the aging response of the alloy by alleviating the uneven distribution of Sc and Zr. Nevertheless, the micro-alloyed elements exceeded the solid solubility limit in the Al-1.75wt%Fe-1.25wt%Ni-0.2wt%Sc-0.2wt%Zr alloy, and their strengthening effect has ever achieved the best prospect. Finally, both Sc and Zr contents were reduced simultaneously, and the aging response of the Al-1.75wt%Fe-1.25wt%Ni-0.15wt%Sc-0.1wt%Zr alloy was improved by optimized heat treatment. The underlying mechanisms for this alloy design and the corresponding microstructure-mechanical property relationship were analytically discussed.

Converged DNA Damage Response Renders Human Hepatocellular Carcinoma Sensitive to CDK7 Inhibition.

Hepatocellular carcinoma (HCC) is a lethal malignancy with high mortality. The inhibition of cyclin-dependent kinase 7 (CDK7) activity has shown therapeutic efficacy in HCC. However, the underlying molecular mechanisms remain elusive. Here, we show that three HCC lines, HepG2, Hep3B, and SK-Hep-1, were highly susceptible to the CDK7 inhibitor THZ1. In mouse models, THZ1 effectively reduced HepG2 tumor growth and tumor weight. THZ1 arrested cell cycle and triggered MYC-related apoptosis in HepG2. To evaluate how MYC protein levels affected THZ1-induced apoptotic cell death, we overexpressed MYC in HepG2 and found that exogenously overexpressed MYC promoted cell cycle progression and increased cells in the S phase. THZ1 drastically engendered the apoptosis of MYC-overexpressing HepG2 cells in the S and G2/M phases. Importantly, transcription-inhibition-induced apoptosis is associated with DNA damage, and exogenous MYC expression further enhanced the THZ1-induced DNA damage response in MYC-overexpressing HepG2 cells. Consistently, in the HepG2 xenografts, THZ1 treatment was associated with DNA-damage-induced cell death. Together, our data indicate that the converged effect of MYC-promoted cell cycle progression and CDK7 inhibition by THZ1 confers the hypersensitivity of HCC to DNA-damage-induced cell death. Our findings may suggest a new therapeutic strategy of THZ1 against HCC.

An Improved K-Means Algorithm Based on Evidence Distance.

The main influencing factors of the clustering effect of the k-means algorithm are the selection of the initial clustering center and the distance measurement between the sample points. The traditional k-mean algorithm uses Euclidean distance to measure the distance between sample points, thus it suffers from low differentiation of attributes between sample points and is prone to local optimal solutions. For this feature, this paper proposes an improved k-means algorithm based on evidence distance. Firstly, the attribute values of sample points are modelled as the basic probability assignment (BPA) of sample points. Then, the traditional Euclidean distance is replaced by the evidence distance for measuring the distance between sample points, and finally k-means clustering is carried out using UCI data. Experimental comparisons are made with the traditional k-means algorithm, the k-means algorithm based on the aggregation distance parameter, and the Gaussian mixture model. The experimental results show that the improved k-means algorithm based on evidence distance proposed in this paper has a better clustering effect and the convergence of the algorithm is also better.

Effects of postoperative adjuvant radiotherapy on stage IIIA-N2 non-small cell lung cancer and prognostic analysis.

PURPOSE: We aimed to explore the efficacy and safety of postoperative adjuvant radiotherapy in the treatment of non-small cell lung cancer (NSCLC) (stage IIIA-N2), and to analyze the influencing factors for the prognosis of patients. METHODS: A total of 142 patients with NSCLC (stage IIIA-N2) were collected for retrospective analysis. Postoperative adjuvant radiotherapy was performed in 71 cases (Radiotherapy group), while it was not conducted in the remaining 71 cases (Control group). The survival status of patients was recorded during follow-up. Moreover, the possible influencing factors for the prognosis of patients were analyzed. RESULTS: The median survival time was 34.7±5.4 months and 31.9±4.9 months, the 5-year overall survival (OS) rate was 32.4% and 26.8%, and the 5-year progression-free survival (PFS) rate was 25.4% and 12.7%, respectively, in the Radiotherapy group and the Control group. The 5-year OS was significantly correlated with smoking history, tumor T stage, ratio of positive lymph nodes, number of cycles of postoperative chemotherapy, and whether postoperative adjuvant radiotherapy was combined. Moreover, tumor T stage, ratio of positive lymph nodes and whether adjuvant radiotherapy was combined were independent influencing factors for postoperative OS of patients. The lower tumor T stage, lower ratio of positive lymph nodes and adjuvant radiotherapy combined corresponded to the higher OS rate. CONCLUSIONS: Postoperative adjuvant radiotherapy is safe and feasible in the treatment of NSCLC (stage IIIA-N2), which can increase the survival of patients and the local control rate of tumors. Patients with a lower tumor T stage and a lower ratio of positive lymph nodes have higher survival rates.

Mitogen-activated protein kinase inhibition-induced modulation of epidermal growth factor receptor signaling in human head and neck squamous cell carcinoma.

BACKGROUND: Epidermal growth factor receptor (EGFR) overexpression is one of the most notable characteristics in head and neck squamous cell carcinoma (HNSCC). The MAPK kinase (MEK) inhibitor trametinib has shown efficacy to treat HNSCC; however, the molecular mechanism remains unclear. METHODS: HNSCC lines, mouse models, Western blot, and flow cytometry were employed to analyze the anticancer effects of trametinib. RESULTS: The JHU-011, JHU-022, and JHU-029 HNSCC cells with different genetic alterations were highly susceptible to trametinib. Trametinib effectively reduced EGFR expression, which was accompanied by the reduction of pro-survival protein MYC, and the increased expression of a MYC-targeted cyclin-dependent kinase inhibitor p27kip1 and pro-apoptotic protein BIM. Trametinib resulted in G1 arrest of the cells, markedly reduced cell numbers in S phase, and significantly increased apoptosis. In mouse models, trametinib strongly inhibited tumors growth. CONCLUSIONS: The MAPK-ERK signaling inhibition by trametinib may target EGFR and the downstream proteins against HNSCC.

Inhibition of MEK suppresses hepatocellular carcinoma growth through independent MYC and BIM regulation.

BACKGROUND: Hepatocellular carcinoma (HCC) is an aggressive malignancy. In HCC, mitogen-activated protein kinase (MAPK) signaling is overactivated. The MAPK kinase (MEK) inhibitor trametinib has been approved to treat several types of advanced cancers with a BRAF mutation. Herein, we examined whether trametinib has efficacy against HCC. METHODS: The effects of trametinib on cell viability, proliferation and tumor growth were assessed in HCC-derived cell lines and mouse xenograft models. Western blot analysis and immunohistochemistry were used to identify key regulators critical for HHC cell proliferation and tumor growth. RESULTS: We found that trametinib dose-dependently inhibited the viability and proliferation of HCC cells. We also found that a strong suppression of MEK by trametinib downregulated the pro-survival protein MYC, but upregulated the pro-apoptotic protein BIM. This dual differential regulation of MYC and BIM was found to be accompanied by upregulation of a MYC-targeted cyclin dependent kinase inhibitor, p27kip1 (p27), and an apoptosis marker, cleaved poly (ADP ribose) polymerase 1 (PARP), indicating a concurrent modulation of cell cycle- and apoptosis-related pathways. Importantly, we found that MYC overexpression did not block increased BIM in trametinib-treated HCC cells, indicating that MAPK signaling independently regulates MYC and BIM. Finally, we found that trametinib in vivo inhibited HepG2 xenograft tumor growth and attenuated tumor invasion into surrounding tissues. Consistent with the in vitro findings, MYC expression was found to be reduced, while p27 expression was found to be elevated, and BIM expression and cleaved PARP levels were found to be increased in trametinib-treated xenograft tumors. CONCLUSIONS: Collectively, our data indicate that trametinib exhibits efficacy in treating HCC cells via distinct regulation of the MYC and BIM pathways. As such, targeting MEK to block MAPK signaling with trametinib may provide novel treatment opportunities for HCC.

The rs10229583 polymorphism near paired box gene 4 is associated with gestational diabetes mellitus in Chinese women.

Objective The rs10229583 polymorphism near paired box gene 4 ( PAX4) is associated with insulin resistance and type 2 diabetes. Mutations in the PAX4 gene may be associated with impaired differentiation/development of pancreatic islet beta cells during fetal development and, consequently, a compromised insulin response to high blood glucose. To ascertain whether this polymorphism plays a role in gestational diabetes mellitus (GDM), we investigated the genotypic and allele frequency differences between GDM and normal pregnancies. Methods A total of 310 GDM and 440 normal pregnancies were evaluated. Allele and genotype frequencies of rs10229583 were determined for all participants with Sanger sequencing and SNaPshot. Association of the allele and genotypes of the single nucleotide polymorphism with the disease was analyzed using Pearson's χ2 test and OR (odds ratio). Results The G allele was more frequent in patients with GDM compared with controls (OR = 1.47, 95% confidence interval (CI): 1.12-1.939). The GG genotype frequency of rs10229583 was significantly different between subjects with GDM and normal controls (OR = 1.411, 95% CI: 1.032-1.928). The OR of the GA + GG genotype was 3.182 (95% CI: 1.294-7.826) for patients with GDM compared with controls. Conclusion The present study suggests that rs10229583 is associated with GDM.

Simple quantitative PCR analysis for allelic Pten loss in tumor progression.

We describe a simple method to accurately detect and quantify both Pten mutation and allele-specific loss using allele-specific PCR analysis. Our approach used a heterozygous genomic DNA with one wild-type and one mutant Pten allele as a reference at a single concentration to calculate the percent ratio of the wild-type Pten gene for the detection of allele-specific gene loss. With a standard curve, ratios from PCR data were used to quantitate the wild-type Pten allele copy number loss in tumor specimens. We demonstrate the utility of our approach to calculate allele-specific Pten loss during tumor progression and show that our approach generates quantitative data that are comparable to those obtained from digital droplet PCR. As a method to detect both mutation and allele-specific gene loss, our approach is less subject to the variability of sample amount that are often very limited in clinical analysis. Since conventional PCR is easy to be carried out, our method simplifies the workflow in any laboratory and would provide significant advantages for simplicity to quantify allele-specific gene loss.

Probe-free allele-specific copy number detection and analysis of tumors.

Cancer development and progression frequently involve nucleotide mutations as well as amplifications and deletions of genomic segments. Quantification of allele-specific copy number is an important step in characterizing tumor genomes for precision medicine. Despite advances in approaches to high-throughput genomic DNA analysis, inexpensive and simple methods for analyzing complex nucleotide and copy number variants are still needed. Real-time polymerase chain reaction (PCR) methods for discovering and genotyping single nucleotide polymorphisms are becoming increasingly important in genetic analysis. In this study, we describe a simple, single-tube, probe-free method that combines SYBR Green I-based quantitative real-time PCR and quantitative melting curve analysis both to detect specific nucleotide variants and to quantify allele-specific copy number variants of tumors. The approach is based on the quantification of the targets of interest and the relative abundance of two alleles in a single tube. The specificity, sensitivity, and utility of the assay were demonstrated in detecting allele-specific copy number changes critical for carcinogenesis and therapeutic intervention. Our approach would be useful for allele-specific copy number analysis or precise genotyping.