A Study on the Effect of IL-17A on Phenotypic Transformation of Fibroblasts in Bleomycin-Induced Pulmonary Fibrosis in Mice and Its Mechanism

Objective: In this study, lung fibroblasts were cultured and identified in mice lung fiber model with bleomycin. Under the induction of IL-17A, lung fibroblasts were gradually transformed into myofibroblasts in pulmonary fibrosis, and the specific induction effect of IL-17A in pulmonary fibrosis was analyzed, which could provide ideas for the prevention and treatment of clinical pulmonary fibrosis. Methods: To investigate the transcriptional expression of bleomycin-induced fractional pulmonary fibrosis in different pulmonary fibrosis processes. The 14-day mice model was taken as the research object, and the pulmonary fibrosis model was established by induction of myogenesis. After 14 days of modeling, lung tissue was removed, and after centrifugation and repeated adherent treatment, lung fibroblasts could be cultured at the origin. After three generations of culture, the morphological changes of lung fibroblasts could be observed under a microscope. Indirect immunofluorescence was used to establish the expression of vimentin, and IL-17 was used to stimulate primary cultured lung fibroblasts to detect the expression and specific localization of a-SMA in cells. Western blotting was used to stimulate the expression of lung fibroblast protein by IL-17A at different time points. Results: The typical characteristics of primary culture lung fibroblasts were obtained. After purification and culture, lung fibroblasts were obtained in morphology. The morphology of the 3rd and 4th generation cells was relatively uniform, showing long carboxyform. 1-2 nucleoli can be observed by microscope, which have distinct cell boundary and are lined up like fish schools. The results of indirect immunofluorescence showed that the vimentin staining in the third generation cells was positive, and the plasma was dark red. There were collagenous fibrous septa between the cells, which might make them develop into lung fibroblasts. A-SMA immunofluorescence results showed that in the absence of IL-17A induction, A-SMA signal was relatively weak in the lung fibroblasts of the control group and was in the cytoplasm, while after IL-17A induction, A-SMA signal was stronger in the lung fibroblasts of mice and the whole cells presented spindle structure. Western bletting showed that lung fibroblasts were stimulated by IL-17 in the 0h group. Compared with the 1h, 2h, and 4h groups, the expression of A-SMA in lung fibroblasts was significantly increased in the 1h, 2h, and 4h groups. The fibroblasts were very low in the 2h and 4h groups. There was no significant difference in the expression of AS MA signal. Compared with 0h, protein contents of p-IKB-a and p-p65 were higher in lung fibroblasts at 1h, 2h and 4h. Protein expressions of Acti, 1P6, IKB-a and P65 were different in lung fibroblasts, but there was no significant difference. However, there was no significant statistical difference in the expression of these proteins in lung fibroblasts at different times. Conclusion: By differential centrifugation and repeated adhesion, bleomycin-induced lung fibroblasts can be isolated and purified, and more cell production can be obtained. The staining vimentin was strongly positive after identification by indirect immunofluorescence. The stimulation of IL-17A could gradually transform non-fibroblasts into myofibroblasts and play an important role in pulmonary fibrosis. Therefore, through experimental studies, it was found that IL-17A stimulated F-kB signal and then increased the expression of P-IKB-a and P-P65 proteins, and transformed non-phosphorylated proteins into phosphorylated proteins, thus transforming lung fibroblasts into myofibroblasts and playing a role in pulmonary fibrosis

Temporal variability in composition and fluxes of Yellow River particulate organic matter

This study examines temporal variations of the abundance and carbon isotopic characteristics of particulate organic carbon (POC) and specific-source compounds in the context of hydrological variability in the Yellow River. The content and bulk carbon isotopic characteristics (13C and 14C) of POC were relatively uniform over the hydrologic (seasonal) cycle. We attribute these temporally invariant geochemical characteristics to the dominant contribution of loess material to the suspended particulate matter (SPM). In contrast, molecular-level signals revealed that hydrologic conditions exert a significant influence on the proportional contributions of petrogenic and especially fresh plant-derived OC, while pre-aged soil OC is mobilized via deeper erosion processes (e.g., gully erosion, mudslides) and is independent of hydrodynamics and surface runoff. A coupled biomarker-isotope mixing model was applied to estimate the time-varying supply of contemporary/modern biomass, pre-aged soil, and fossil OC components to Chinese marginal seas from the Yellow River. We found that natural (e.g., precipitation) and human-induced (e.g., water and sediment regulation) variations in hydrological regime strongly influence the flux with the magnitude of the corresponding annual fluxes of POC ranging between 0.343 ± 0.122 Mt yr−1 and 0.581 ± 0.213 Mt yr−1, but less strongly infleunce proportions of the different OC constituents. Inter-annual differences in pre-aged soil and fossil OC fluxes imply that extreme climate events (e.g., floods) modulate the exhumation and export of old carbon to the ocean, but the OC homogeneity in the pre-aged mineral soil-dominated watersheds facilitates robust predictions in terms of OC transport dynamics in the past (sediment cores) and in the future

Complexation of Z-ligustilide with hydroxypropyl-β-cyclodextrin to improve stability and oral bioavailability

To improve the stability and oral bioavailability of Z-ligustilide (LIG), the inclusion complex of LIG with hydroxypropyl-β-cyclodextrin (HP-β-CD) was prepared by the kneading method and characterized by UV-Vis spectroscopy, differential thermal analysis (DTA) and Fourier transform infrared (FTIR) spectroscopy. LIG is capable of forming an inclusion complex with HP-β-CD and the stoichiometry of the complex was 1:1. Stability of the inclusion complex against temperature and light was greatly enhanced compared to that of free LIG. Further, oral bioavailability of LIG and the inclusion complex in rats were studied and the plasma drug concentration-time curves fitted well with the non-compartment model to estimate the absolute bioavailability, which was 7.5 and 35.9 %, respectively. In conclusion, these results show that LIG/HP-β-CD complexation can be of great use for increasing the stability and biological efficacy of LIG

Introgressive hybridization between two close species Siniperca chuatsi and Siniperca kneri (Percomorpharia: Sinipercidae) in the Middle Reaches of the Yangtze River

Siniperca chuatsi and Siniperca kneri are two economically important freshwater fishes endemic to East Asia. Recently, some Siniperca specimens collected from Lake Poyang and Lake Dongting in the middle reaches of the Yangtze River couldn't be clearly identified as they showed intermediate morphological characteristics between S. chuatsi and S. kneri, and some inter-species hybrids were detected by microsatellite loci. To further verify genetic composition of these intermediate individuals, and determine the prevalence and degree of introgression between the two Siniperca species, a large set of high-quality, independent, diagnostic genetic markers were necessarily required. Based on enrichment and sequencing of target genes in sinipercid fishes, 463 loci (FST = 1) between S. chuatsi and S. kneri were selected and verified for species diagnosis. A total of 349 loci with 458 diagnostic SNPs were identified for discriminating S. chuatsi and S. kneri. From those markers, 224 diagnostic SNPs (only one SNP per locus) were selected to identify and categorize 48 specimens with intermediate morphological characters. The results showed that there were 8 specimens identified as hybrids, 8 specimens as S. chuatsi and 32 specimens as S. kneri. NEWHYBRIDS analysis showed that the hybrid offsprings were composed of the first-generation hybrid (2 individuals), first-generation backcross (1 individual), second-generation backcross (1 individual) and fourth-generation backcross (4 individuals), and the backcrossing could happen to both S. chuatsi and S. kneri. These hybrids could occur naturally, or escaped from farmed fish, due to extensive artificial breeding practice in these regions. However, the origin of the introgressive hybridization can't be easily traced. Therefore, some measures for protecting genetic resource of Siniperca speies in the Yangtze River should be enforced, such as assessing genetic background of the cultured stocks, reducing the escapement from farmed fish, and monitoring the trend of introgressive hybridization between Siniperca species in the future

China’s pig relocation in balance

In 2015, the Chinese government banned livestock production in some regions (called non-livestock production regions, NLPRs) to control surface water pollution near vulnerable water bodies. In total, 90,000 NLPRs had been established by 2017, covering a land area of 0.82 million km2 and shutting down 0.26 million pig farms1. As a consequence, the number of slaughtered pigs decreased by 46 million head yr–1 between 2014 and 2017. The NLPRs policy is globally unprecedented in terms of the geographical area and number of farms affected, as well as its implementation speed. The NLPRs policy has reduced pork self-sufficiency in some provinces by up to 40% (ref. 2). However, it is unclear which farms and regions may take over the market share

Normal spermatogenesis in Fank1 (fibronectin type 3 and ankyrin repeat domains 1) mutant mice

Background The fibronectin type 3 and ankyrin repeat domains 1 gene, Fank1, is an ancient, evolutionarily conserved gene present in vertebrates. Short-hairpin RNA (shRNA)-based knockdown transgenic mice have oligospermia caused by an increase in apoptotic germ cells. In this study, we investigated the in vivo function of Fank1. Methods In this study, we generated Fank1-knockout mice using the CRISPR/Cas9 system. We then investigated the phenotype and in vivo function of Fank1. Testes and epididymis tissues were analyzed by histological and immunofluorescence staining. Apoptotic cells were analyzed in terminal deoxynucleotidyl transferase dUTP nick end-labeling assays. Fertility and sperm counts were also evaluated. The GTEx database were used to assess gene expression quantitative trait loci and mRNA expression of candidate genes and genes neighboring single nucleotide polymorphisms was analyzed by quantitative RT-PCR. Results In contrast to the Fank1-knockdown model, no significant changes in epididymal sperm content and the number of apoptotic cells were observed in Fank1−/− homozygotes. In addition, a different pattern of Dusp1, Klk1b21 and Klk1b27 mRNA expression was detected in Fank1-knockout testis. These results reveal differences in the molecular changes between Fank1-knockdown mice and Fank1-knockout mice and provide a basic resource for population genetics studies

Causal associations of gut microbiota and metabolites on sepsis: a two-sample Mendelian randomization study

BackgroundSepsis stands as a dire medical condition, arising when the body’s immune response to infection spirals into overdrive, paving the way for potential organ damage and potential mortality. With intestinal flora’s known impact on sepsis but a dearth of comprehensive data, our study embarked on a two-sample Mendelian randomization analysis to probe the causal link between gut microbiota and their metabolites with severe sepsis patients who succumbed within a 28-day span.MethodsLeveraging data from Genome-wide association study (GWAS) and combining it with data from 2,076 European descendants in the Framingham Heart Study, single-nucleotide polymorphisms (SNPs) were employed as Instrumental Variables (IVs) to discern gene loci affiliated with metabolites. GWAS summary statistics for sepsis were extracted from the UK Biobank consortium.ResultsIn this extensive exploration, 93 distinct genome-wide significant SNPs correlated with gut microbial metabolites and specific bacterial traits were identified for IVs construction. Notably, a substantial link between Coprococcus2 and both the incidence (OR of 0.80, 95% CI: 0.68-0.94, P=0.007) and the 28-day mortality rate (OR 0.48, 95% CI: 0.27-0.85, P=0.013) of sepsis was observed. The metabolite α-hydroxybutyrate displayed a marked association with sepsis onset (OR=1.08, 95% CI: 1.02-1.15, P=0.006) and its 28-day mortality rate (OR=1.17, 95% CI: 1.01-1.36, P=0.029).ConclusionThis research unveils the intricate interplay between the gut microbial consortium, especially the genus Coprococcus, and the metabolite α-hydroxybutyrate in the milieu of sepsis. The findings illuminate the pivotal role of intestinal microbiota and their metabolites in sepsis’ pathogenesis, offering fresh insights for future research and hinting at novel strategies for sepsis’ diagnosis, therapeutic interventions, and prognostic assessments

Blastocyst quality and reproductive and perinatal outcomes : a multinational multicentre observational study

Funding H.Z. is supported by a Monash Research Scholarship. B.W.J.M. is supported by an NHMRC Investigator grant (GNT1176437). R.W. is supported by an NHMRC Emerging Leadership Investigator grant (2009767).Peer reviewedPublisher PD