[Serum metabolomics in latent pneumoconiosis tuberculosis patients based on ultra performance liquid chromatography tandem quadrupole time of flight mass spectrometry].

Objective: To explore the non-target metabonomics of serum in worker's pneumoconiosis (CWP) patients with latent tuberculosis and the biomarkers of latent tuberculosis infection of pneumoconiosis. Methods: In December 2018, 39 CWP inpatients from a hospital in Beijing were taken as subjects. The subjects were screened for latent tuberculosis using the in vitro release test of mycobacterium tuberculosis-interferon (IGRAs) test. According to the screening results, 21 positive patients with latent tuberculosis infection were selected as the latent tuberculosis group of pneumoconiosis. While 18 negative patients with CWP alone were selected as the pneumoconiosis group. Polarity components of metabolites were analyzed by UPLC-QTOF/MS. The data was processed with Progenesis QI software for multidimensional statistical analysis. Identification of structure of differential metabolites were matched through accurate mass and secondary mass spectrum. Searching the Human Metabolome Database (HMDB) , differential metabolites were imported into MetaboAnalyst 4.0 to analyze the metabolic pathways. Results: All 42 differential metabolites were screened out. Excepted for exogenous metabolites, 14 endogenous differential metabolites were identified. Compared with the pneumoconiosis group, 6 metabolites including PC [18∶4 (6Z, 9Z, 12Z, 15Z) /P-18∶1 (11Z) ], 3-Oxododecanoyl-CoA in the latent tuberculosis group were up-regulated, while 8 metabolites including the Stearoyl-CoA, (2S) -Pristanoyl-CoA were down-regulated. These results might be related to lipid, fatty acid and arachidonic acid metabolism pathways. Conclusion: There are significant differences in serum metabonomics between the patients with latent tuberculosis of pneumoconiosis and the patients with ordinary pneumoconiosis, which provide a reference for the study of biomarkers for the diagnosis of latent tuberculosis infection of pneumoconiosis.

The SNP-set based association study identifies ITGA1 as a susceptibility gene of attention-deficit/hyperactivity disorder in Han Chinese.

Genome-wide association studies, which detect the association between single-nucleotide polymorphisms (SNPs) and disease susceptibility, have been extensively applied to study attention-deficit/hyperactivity disorder (ADHD), but genome-wide significant associations have not been found yet. Genetic heterogeneity and insufficient genomic coverage may account for the missing heritability. We performed a two-stage association study for ADHD in the Han Chinese population. In the discovery stage, 1033 ADHD patients and 950 healthy controls were genotyped using both the Affymetrix Genome-Wide Human SNP Array 6.0 and the Illumina Infinium HumanExome BeadChip. The genotyped SNPs were combined to generate a powerful SNP set with better genomic coverage especially for the nonsynonymous variants. In addition to the association of single SNPs, we collected adjacent SNPs as SNP sets, which were determined by either genes or successive sliding windows, to evaluate their synergetic effect. The candidate susceptibility SNPs were further replicated in an independent cohort of 1441 ADHD patients and 1447 healthy controls. No genome-wide significant SNPs or gene-based SNP sets were found to be associated with ADHD. However, two continuous sliding windows located in ITGA1 (P-value=8.33E-7 and P-value=8.43E-7) were genome-wide significant. The quantitative trait analyses also demonstrated their association with ADHD core symptoms and executive functions. The association was further validated by follow-up replications for four selected SNPs: rs1979398 (P-value=2.64E-6), rs16880453 (P-value=3.58E-4), rs1531545 (P-value=7.62E-4) and rs4074793 (P-value=2.03E-4). Our results suggest that genetic variants in ITGA1 may be involved in the etiology of ADHD and the SNP-set based analysis is a promising strategy for the detection of underlying genetic risk factors.

DNA damage-induced sustained p53 activation contributes to inflammation-associated hepatocarcinogenesis in rats.

The tumor suppressor p53 has an important role in inducing cell-intrinsic responses to DNA damage, including cellular senescence or apoptosis, which act to thwart tumor development. It has been shown, however, that senescent or dying cells are capable of eliciting inflammatory responses, which can have pro-tumorigenic effects. Whether DNA damage-induced p53 activity can contribute to senescence- or apoptosis-associated pro-tumorigenic inflammation is unknown. Recently, we generated a p53 knock-out rat via homologous recombination in rat embryonic stem cells. Here we show that in a rat model of inflammation-associated hepatocarcinogenesis, heterozygous deficiency of p53 resulted in attenuated inflammatory responses and ameliorated hepatic cirrhosis and tumorigenesis. Chronic administration of hepatocarcinogenic compound, diethylnitrosamine, led to persistent DNA damage and sustained induction of p53 protein in the wild-type livers, and much less induction in p53 heterozygous livers. Sustained p53 activation subsequent to DNA damage was accompanied by apoptotic rather than senescent hepatic injury, which gave rise to the hepatic inflammatory responses. In contrast, the non-hepatocarcinogenic agent, carbon tetrachloride, failed to induce p53, and caused a similar degree of chronic hepatic inflammation and cirrhosis in wild type and p53 heterozygous rats. These results suggest that although p53 is usually regarded as a tumor suppressor, its constant activation can promote pro-tumorigenic inflammation, especially in livers exposed to agents that inflict lasting mutagenic DNA damage.

Cancer targeting Gene-Viro-Therapy of liver carcinoma by dual-regulated oncolytic adenovirus armed with TRAIL gene.

Liver cancer is a common and aggressive malignancy, but available treatment approaches remain suboptimal. Cancer targeting Gene-Viro-Therapy (CTGVT) has shown excellent anti-tumor effects in a preclinical study. CTGVT takes advantage of both gene therapy and virotherapy by incorporating an anti-tumor gene into an oncolytic virus vector. Potent anti-tumor activity is achieved by virus replication and exogenous expression of the anti-tumor gene. A dual-regulated oncolytic adenoviral vector designated Ad·AFP·E1A·E1B (Δ55) (Ad·AFP·D55 for short thereafter) was constructed by replacing the native viral E1A promoter with the simian virus 40 enhancer/α-fetoprotein (AFP) composite promoter (AFPep) based on an E1B-55K-deleted construct, ZD55. Ad·AFP·D55 showed specific replication and cytotoxicity in AFP-positive hepatoma cells. It also showed enhanced safety in normal cells when compared with the mono-regulated vector ZD55. To improve the anti-hepatoma activities of the virus, the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) gene was introduced into Ad·AFP·D55. Ad·AFP·D55-TRAIL exhibited remarkable anti-tumor activities in vitro and in vivo. Treatment with Ad·AFP·D55-TRAIL can induce both autophagy owing to the Ad·AFP·D55 vector and caspase-dependent apoptosis owing to the TRAIL protein. Therefore, Ad·AFP·D55-TRAIL could be a potential anti-hepatoma agent with anti-tumor activities due to AFP-specific replication and TRAIL-induced apoptosis.

Potent antitumoral efficacy of a novel replicative adenovirus CNHK300 targeting telomerase-positive cancer cells.

PURPOSE: Telomerase reverse transcriptase (hTERT) is the key determinant of telomerase activity and plays a crucial role in cellular immortalization and oncogenesis. It will be a promising target for cancer gene therapy. We constructed a novel replicative adenovirus CNHK300 in which hTERT promoter with three extra E-boxes downstream of the promoter was introduced and used to regulate adenoviral E1a gene, and studied its properties of selective replication in cancer cells and antitumoral activity. METHODS: Luciferase assay was used to detect hTERT promoter activity. The selective replication of CNHK300 in cancer cells was investigated by E1a Western blot and green fluorescent protein (GFP) reporter gene assay. The antitumoral activity of CNHK300 and its toxicity were measured on animal models. RESULTS: Luciferase assay showed that introducing extra E-boxes downstream of hTERT promoter is beneficial to decreasing the promoter activity in normal cells without affecting its strong activity in cancer cells. Experiments in vitro and in vivo demonstrated that CNHK300 can selectively target to hTERT-positive cancer cells and replicate in them, resulting in oncolytic or antitumoral effect. CNHK300 is superior to ONYX-015 in terms of selective replication and oncolytic or antitumoral effect. The toxicity assay showed no signs of toxicity to liver cells even at the higher dosage of CNHK300 in vivo. CONCLUSION: The hTERT promoter-controlled, replication-competent adenovirus CNHK300 is a promising system for targeted cancer gene therapy.

[2 homoharringtonine resistant leukemic cell lines (K562 HHT and L1210 HHT): establishment, characterization and mechanisms of action].

We developed two leukemic cell lines (K562 HHT and L1210 HHT) stably 16.7 fold and 13.4 fold resistant to HHT respectively with which the culture were treated. Both cell lines were also cross-resistant to DOX, VCR, DNR and Mel. Increased expression of MDR1 gene in the both lines was noted. To further evaluate the implications of MDR1 in HHT resistance. We studied the expression of MDR1 in sensitive and HHT-resistant sublines of K562 by ABC with an monoclonal antibody against P170, JSB-1. K562 HHT cells were positive but sensitive cells were negative. Additionally, the increased drug resistance was associated with increased level of expression of alpha and pi class GST gene, but not with increased level of expression of mu class GST gene.