[Research progress on pneumoconiosis markers based on multi-omics analysis].

The etiology of pneumoconiosis is relatively clear, but the pathogenic mechanism is not fully understood, and there is no effective cure for pneumoconiosis. Clarifying the pathogenesis of pneumoconiosis and exploring relevant markers can help screen high-risk groups of dust exposure, and relevant markers can also be used as targets to intervene in the process of pulmonary fibrosis. The in-depth development of genomics, transcriptomics and proteomics has provided a new way to discover more potential markers of pneumoconiosis. In the future, the combination of multi-omics and multi-stage interactive analysis can systematically and comprehensively identify key genes (proteins) , metabolites and metabolic pathways in the occurrence and development of pneumoconiosis, build a core regulatory network, and then screen out sensitive markers related to early diagnosis and treatment of pneumoconiosis. This article summarizes the research progress of pneumoconiosis markers from the perspective of multi-omics, hoping to provide more basic data for the early prevention and diagnosis of pneumoconiosis, pathogenesis research, and therapeutic intervention.

[Study of the association between interleukin-1 polymorphisms and genetic susceptibility of coal workers' pneumoconiosis and silicosis].

Objective: To evaluate the relationship between IL-1 gene polymorphisms and coal workers' pneumoconiosis and silicosis susceptibility. Methods: We searched published full-text from PubMed, Web of Science, CNKI, VIP and Wanfang to collect case-control study on IL-1 gene polymorphisms with coal workers' pneumoconiosis and silicosis susceptibility. Eight articles, including 10 case-control studies were included in our study. All analyses were performed using the Stata version 12.0 software. Results: The IL-1RA (+2018) TC or CC variant genotypes were associated with coal workers' pneumoconiosis and silicosis risk (OR=1.65, 95%CI: 1.11-2.46) . In further stratified analyses, the IL-1RA (+2018) TC or CC variant genotypes were associated with an increased silicosis risk (OR=2.07, 95%CI: 1.45-2.95) , which were also associated with increased coal workers' pneumoconiosis and silicosis risk in Caucasians (OR=1.74, 95%CI: 1.22-2.47) . No significant association between IL-1ß (+3953) , IL-1ß (-511) , IL-1α (+4845) and coal workers' pneumoconiosis and silicosis risk was found either in the overall study or in the stratified analysis. Conclusion: These findings suggested that IL-1RA (+2018) may modify coal workers' pneumoconiosis and silicosis susceptibility. Further replication studies with large sample sizes are warranted to re-evaluate the relationship between IL-1RA (+2018) and coal workers' pneumoconiosis and silicosis risk.

[Association between a novel regulatory genetic variants and lung cancer risk in Chinese: a two-stage case-control study].

Objective: Regulatory quantitative trait loci (regQTL) theory can help to evaluate the regulation function of single nucleotide polymorphisms (SNPs) on crucial biological signals from a three-dimensional perspective. The aim of this study was to investigate the effect of these regQTL-SNPs on the susceptibility of lung cancer. Methods: Based on the regQTL theory, using the database of identified lung cancer regQTL-SNPs, we screened the SNPs that may function as regQTL in the reported susceptible regions of lung cancer by genome-wide association study(GWAS), and a two-stage case-control study was conducted (screening stage: 2 331 lung cancer cases and 3 077 healthy controls; validation stage: 626 lung cancer cases and 667 healthy controls) to definite the association of related regQTL-SNPs with the susceptibility of lung cancer. Results: A total of 8 regQTL-SNPs were screened in the reported susceptible regions of lung cancer by GWAS. Among which, 3 SNPs were significantly associated with the risk of lung cancer (P<0.05) in the screening stage. Further validation results indicated that the variant T allele of rs6998591 in ADRA1A was significantly associated with increased risk of lung cancer (additive model: OR=1.33, 95%CI:1.01-1.74, P=0.040). In addition, the variant G allele of rs11202916 in ACTA2 was significantly associated with decreased risk of lung cancer (recessive model: OR=0.71, 95%CI:0.52-0.96, P=0.026). Stratified analysis indicated that the variant T allele of rs6998591 significantly increased lung squamous cell carcinoma risk (additive model: OR=1.53, 95%CI: 1.01-2.32, P=0.043), while the variant G allele of rs11202916 significantly decreased lung adenocarcinoma risk (additive model: OR=0.83, 95%CI: 0.69-0.98, P=0.031). Gene-environment interaction analysis indicated that the risk of developing lung cancer increased by 235% in smoking individuals carrying rs6998591 variant T allele compared with those non-smoking individuals carrying no rs6998591 variant T allele(OR=3.35,95%CI:2.10-5.34,P<0.001). Conclusion: There are two regQTL-SNPs that could significantly affect the susceptibility of lung cancer in the GWAS reported susceptible regions of lung cancer.

Transcription of the human hepatic lipase gene is modulated by multiple negative elements in HepG2 cells.

The expression of the hepatic lipase (HL) gene is highly tissue specific. In order to identify cis-acting elements which regulate the expression of this gene in the liver, multiple deletion mutants of the 5'-flanking region of the HL gene fused to the human growth hormone gene were transfected in HepG2 cells, which normally produce HL. Transient expression assays indicated the presence of negative (at nucleotides (nt) -1576(/)-1342 and -623(/)-407) and positive (at nt -1862(/)-1576 and -50(/)-9) regulatory elements. Transfection of HeLa cells, which do not produce HL, with the same deletion constructs resulted in a similar pattern of promoter activities. However, additional negative (nt -138/-50) and positive (nt -407(/)-138) elements were found. DNase I footprint analysis of the proximal and distal HLpromoter sequences with HepG2 and HeLa cell nuclear extracts identified seven protected regions: A, nt -1540(/)-1527; B, -1505(/)-1473; C, -1467(/)-1460; D, -592(/)-577; E, -565(/)-545; F, -234(/)-220; and G, -70(/) -48. Sites A, B, C, D and E were located within regions containing negative regulatory elements. In order to determine which nuclear factor interacts with the negative elements, sites B, D and E were mutated and the effects of mutation on competition in a gel retardation assay and on promoter activity were studied. When the binding motif for AP1 in sites B, D and E was mutated, the specific DNA-protein complexes were not competed with the mutant oligonucleotides and promoter activity increased twofold. The magnitude of the increase is less than expected from the deletion analysis, and simultaneous mutations did not cause further increase in promoter activity, which suggests that other sites are involved in this negative modulation. These results suggest that the transcription of the HLgene in HepG2 cells is negatively modulated by multiple cis-acting negative elements and AP1-like nuclear factor may play some role in this modulation.

The effect of pressure on ultrasonic testing sensitivity of hot steel blooms by pressure contact method.

The effect of exerted pressure of the buffer rod on the sensitivity of ultrasonic testing of hot steel blooms by the pressure contact method is discussed. The formula describing their linear relation is introduced. Results show that, at a given temperature, sensitivity increases linearly with pressure until a certain pressure is reached where sensitivity is rather insensitive to pressure variations. However, this pressure region is not always used. The proper choice of exerted pressure for practical use is analyzed.

Partial structure of the mouse glucokinase gene.

A complementary DNA for glucokinase (GK) was cloned from mouse liver total RNA by a combination of the polymerase chain reaction (PCR) and mouse liver cDNA library screening. Liver- and beta-cell-specific exons 1 were isolated by PCR using mouse and rat genomic DNAs. These clones were then used to screen a mouse genomic library; three genomic clones were isolated and characterized. The mouse GK gene spans over 20 kb, containing 11 exons including a liver- or beta-cell-specific exon 1, which encodes a tissue-specific 15-aa peptide at the N-terminus of the protein. Both types of GK contain 465 amino acid residues. The predicted amino acid sequence of mouse beta-cell-specific GK showed 98 and 96% identity to the rat and human enzymes, respectively; the corresponding values are 98 and 95%, respectively, for the liver-specific GK. Several transcription factor-binding consensus sequences are identified in the 5' flanking region of the mouse GK gene.

Cytotoxic sterols from the soft coral Nephthea erecta.

Two new cytotoxic sterols, 24-methylcholesta-5,24(28)-diene-3beta, 15beta,19-triol (1) and 24-methylcholesta-5,24(28)-diene-3beta, 19-diol-7-one (2), as well as four cytotoxic sterols, 24-methylcholesta-5,24(28)-diene-3beta,19-diol (3), 24-methylcholesta-5,24(28)-diene-3beta,19-diol-7beta-mono aetate (4), 24-methylcholesta-5,24(28)-diene-3beta,7beta,19-triol (5), and 24-methylcholesta-24(28)-ene-3beta,5alpha,6beta,19-tet raol (6), have been isolated from the soft coral Nephthea erecta. The structures of compounds 1 and 2 were determined by spectral analysis.

The nucleotide sequence of surface antigen gene of hepatitis B virus subtype adr.

The nucleotide sequence of the XhoI-BamHI fragment (1279bp), which contains the surface antigen gene (S gene) of HBVadr, was determined by Maxam and Gilbert's method. By comparing the differences both of the nucleotide sequence in the S gene and its coded amino acid sequence between adr and those reported for adw, ayw and adyw, some new variation sites were discovered. The differences were mainly distributed in the two hydrophilic regions. However, at those sites which might show biological function, there were no variations among different subtypes, they are relatively conservative in heredity and evolution. Comparing the variation of the nucleotide sequence in the S gene region with that in the non-S gene region, it is shown that the frequency of variation in the non-S gene region doubled that in the S gene region. The S gene region is more conservative.

Mouse very-low-density-lipoprotein receptor (VLDLR) cDNA cloning, tissue-specific expression and evolutionary relationship with the low-density-lipoprotein receptor.

The very-low-density-lipoprotein receptor (VLDLR) is a recently described lipoprotein receptor that shows considerable similarity to the low-density-lipoprotein receptor (LDLR). This receptor has been suggested to be important for the metabolism of apoprotein-E-containing triacylglycerol-rich lipoproteins, such as very-low-density-lipoprotein (VLDL), beta-migrating VLDL and intermediate-density lipoprotein. cDNA clones that code for the VLDLR were isolated from a mouse heart cDNA library. The deduced amino acid sequence predicts a mature protein of 846 amino acids preceded by a 27-residue signal peptide. Three mRNA species for the VLDLR with sizes of 3.9, 4.5 and 7.9 kilobases were present in high concentration in heart and muscle, which utilize triacylglycerols as an energy source. VLDLR mRNA is also detected in decreasing amounts in kidney, brain, ovary, testis, lung and adipose tissue. It is essentially absent in liver and small intestine. The amino acid sequence of the VLDLR is highly conserved among rabbit, human and mouse. VLDLR contains five structural domains very similar to those in LDLR, except that the ligand-binding domain in VLDLR has an eightfold repeat instead of a sevenfold repeat in LDLR. Sequence conservation among animal species is much higher for the VLDLR than the LDLR. Sequences of the VLDLR from three vertebrate species and the LDLR from five vertebrate species were aligned and a phylogenetic tree was reconstructed. Although both receptors contain five domains and share amino acid sequence similarity, our computations showed that they diverged before the divergence between mammals and amphibians. In addition, sequence comparison of both receptor sequences suggests that the rabbit is evolutionarily closer to man than to the mouse. These results are consistent with the hypothesis that the VLDLR and the LDLR have evolved from a common ancestral gene to play distinct roles in lipoprotein metabolism and that the metabolic handling of triacylglycerol by the body via the VLDLR is a highly conserved mechanism.

The complete nucleotide sequence of the cloned DNA of hepatitis B virus subtype adr in pADR-1.

The complete nucleotide sequence of the cloned hepatitis B virus DNA subtype adr in pADR-1 was determined by Maxam and Gilbert's method. It is 3215 base pairs in size, which is 27 bp longer than the sequence of the adr pHBr330, as reported by Ono et al. The nucleotide difference between pADR-1 and adr pHBr330 is about 2% while those between pADR-1 and adw as well as ayw are 9.3% and 9.7% respectively. In this paper, the heterogeneity and homogeneity of the S gene, the C gene and the other coding regions in pADR-1 and in the other subtypes are compared and discussed.

Essential role of BETA2/NeuroD1 in development of the vestibular and auditory systems.

BETA2/NeuroD1 is a bHLH transcription factor that is expressed during development in the mammalian pancreas and in many locations in the central and peripheral nervous systems. During inner ear ontogenesis, it is present in both sensory ganglion neurons and sensory epithelia. Although studies have shown that BETA2/NeuroD1 is important in the development of the hippocampal dentate gyrus and the cerebellum, its functions in the peripheral nervous system and in particular in the inner ear are unclear. Mice carrying a BETA2/NeuroD1 null mutation exhibit behavioral abnormalities suggestive of an inner ear defect, including lack of responsiveness to sound, hyperactivity, head tilting, and circling. Here we show that these defects can be explained by a severe reduction of sensory neurons in the cochlear-vestibular ganglion (CVG). A developmental study of CVG formation in the null demonstrates that BETA2/NeuroD1 does not play a primary role in the proliferation of neuroblast precursors or in their decision to become neuroblasts. Instead, the reduction in CVG neuron number is caused by a combination both of delayed or defective delamination of CVG neuroblast precursors from the otic vesicle epithelium and of enhanced apoptosis both in the otic epithelium and among those neurons that do delaminate to form the CVG. There are also defects in differentiation and patterning of the cochlear duct and sensory epithelium and loss of the dorsal cochlear nucleus. BETA2/NeuroD1 is, thus, the first gene to be shown to regulate neuronal and sensory cell development in both the cochlear and vestibular systems.