RESUMEN
Activated hepatic stellate cells (HSCs) play a detrimental role in liver fibrosis progression. Natural killer (NK) cells are known to selectively recognize abnormal or transformed cells via their receptor activation and induce target cell apoptosis and, therefore, can be used as a potential therapeutic strategy for liver cirrhosis. In this study, we examined the therapeutic effects of NK cells in the carbon tetrachloride (CCl4)-induced liver cirrhosis mouse model. NK cells were isolated from the mouse spleen and expanded in the cytokine-stimulated culture medium. Natural killer group 2, member D (NKG2D)-positive NK cells were significantly increased after a week of expansion in culture. The intravenous injection of NK cells significantly alleviated liver cirrhosis by reducing collagen deposition, HSC marker activation, and macrophage infiltration. For in vivo imaging, NK cells were isolated from codon-optimized luciferase-expressing transgenic mice. Luciferase-expressing NK cells were expanded, activated and administrated to the mouse model to track them. Bioluminescence images showed increased accumulation of the intravenously inoculated NK cells in the cirrhotic liver of the recipient mouse. In addition, we conducted QuantSeq 3' mRNA sequencing-based transcriptomic analysis. From the transcriptomic analysis, 33 downregulated genes in the extracellular matrix (ECM) and 41 downregulated genes involved in the inflammatory response were observed in the NK cell-treated cirrhotic liver tissues from the 1532 differentially expressed genes (DEGs). This result indicated that the repetitive administration of NK cells alleviated the pathology of liver fibrosis in the CCl4-induced liver cirrhosis mouse model via anti-fibrotic and anti-inflammatory mechanisms. Taken together, our research demonstrated that NK cells could have therapeutic effects in a CCl4-induced liver cirrhosis mouse model. In particular, it was elucidated that extracellular matrix genes and inflammatory response genes, which were mainly affected after NK cell treatment, could be potential targets.
RESUMEN
The modified zeo-SBR is recommended for a new nitrogen removal process that has a special function of consistent ammonium exchange and bioregeneration of zeolite-floc. Three sets of sequencing batch reactors, control, zeo-SBR, and modified zeo-SBR were tested to assess nitrogen removal efficiency. The control reactor consisted of anoxic-fill, aeration-mixing, settling, and decanting/idle phases, meaning that nitrogen removal efficiency was dependent on the decanting volume in a cycle. The zeo-SBR reactor was operated in the same way as the control reactor, except for daily addition of powdered zeolite in the SBR reactor. The operating order sequences in the zeo-SBR were changed in the modified zeo-SBR. Anoxic-fill phase was followed by aeration-mixing phase in the zeo-SBR, while aeration-mixing phase was followed by anoxic-fill phase in the modified zeo-SBR to carry NH4(+)-N over to the next operational cycle and to reduce total nitrogen concentration in the effluent. In the modified zeo-SBR, nitrification and biological regeneration occurred during the initial aeration-mixing phase, while denitrification and ammonium adsorption occurred in the following anoxic-fill phase. The changed operational sequence in the modified zeo-SBR to adapt the ammonium adsorption and biological regeneration of the zeolite-floc could enhance nitrogen removal efficiency. As a result of the continuous operation, the nitrogen removal efficiencies of the control and zeo-SBR were in 68.5-70.9%, based on the 33% of decanting volume for a cycle. The zeo-SBR showed a consistent ammonium exchange and bio-regeneration in the anoxic-fill and aeration-mixing phases, respectively. Meanwhile, the effluent total nitrogen of the modified zeo-SBR showed 50-60 mg N/L through ammonium adsorption of the zeolite-floc when the influent ammonium concentration was 315 mg N/L, indicating the T-N removal efficiency was enhanced over 10% in the same HRT and SRT conditions as those of control and zeo-SBR reactors. The ammonium adsorption capacity was found to be 6-7 mg NH4(+)-N/g FSS that is equivalent to 40 mg NH4(+)-N/L of ammonium nitrogen removal.
