Effects of chitinase-3-like protein 1 on brain death-induced hepatocyte apoptosis via PAR2-JNK-caspase-3.

Hepatocyte apoptosis is a crucial factor affecting liver quality in brain-dead donors. The identification of key molecular proteins involved in brain-death (BD)-induced hepatocyte apoptosis may help determine an effective method for improving the quality of livers from brain-dead donors. In this study, we used in vivo and in vitro models to investigate the role of chitinase-3-like protein 1 (CHI3L1) in promoting liver cell apoptosis after BD. Chitin was used to inhibit CHI3L1 in a rat model of BD. Macrophage polarization of THP-1 cells and hypoxia/reoxygenation (H/R) of LO-2 cells were used to mimic BD-induced cell stress in liver. We found that CHI3L1 played a vital role in promoting liver cell apoptosis. Six hours after BD, CHI3L1 expression was significantly upregulated in liver macrophages and was associated with BD-induced M1 polarization of these cells. In liver cells cultured under H/R conditions, recombinant CHI3L1 activated the protease-activated receptor 2 (PAR2)/c-June N-terminal kinase (JNK) apoptotic pathway and aggravated apoptosis. Compared with the control group, chitin particles inhibited the expression of CHI3L1 in the liver of brain dead rats, thereby reducing activation of the hepatocyte surface receptor, PAR2, and its downstream JNK/caspase-3 signaling pathway, ultimately reducing hepatocyte apoptosis. In conclusion, our results indicate that CHI3L1 relies on a PAR2/JNK-mediated mechanism to promote BD-induced hepatocyte apoptosis.

[Advances in biosynthesis of indigo in plants].

Natural indigo, as one of the oldest dyes, is also a pivotal dye utilized in cotton fabrics today. A diversity of plants rich in indigo compounds belong to traditional Chinese herbal medicines. Indigo compounds have a variety of biological and pharmacological activities, including anticonvulsant, antibacterial, antifungal, antiviral and anticancer activities. A substantial progress in indigo biosynthesis has been made lately. This paper summarizes the value of indigo from the aspects of cultural history, biosynthetic pathways and the medicinal activities of its related derivatives involved in the pathways. In addition, the latest research advancements in indigo biosynthetic pathways is demonstrated in this paper, which would lay the theoretical foundation for the exploration and utilization of natural indigo.

Novel insights into the apoptosis mechanism of DNA topoisomerase I inhibitor isoliquiritigenin on HCC tumor cell.

The inhibitory effect of DNA topoisomerase (Top I) by isoliquiritigenin(ISO) were investigated and their interaction mechanism was evaluated using methods including UV-vis absorption, fluorescence, coupled with molecular simulation, and using the MTT method of inhibition rate of HCC tumor cell SNU475 proliferation assay, finally, the interaction of ISO with calf thymus DNA was investigated by melting measurements and molecular docking studies. It was found that isoliquiritigenin reversibly inhibited DNA Top I in a competitive manner with the concentrations of ISO resulting in 50% activity lost (IC50) were estimated to be 0.178 ± 0.12 mM. Isoliquiritigenin exhibited a strong ability to quench the intrinsic fluorescence of Top I through a static quenching procedure. The positive values of enthalpy change and entropy change suggested that the binding of isoliquiritigenin to Top I was driven mainly by hydrophobic interactions. The molecular docking results revealed isoliquiritigenin actually interacted with the primary amino acid residues on the active site of Top I, and the detection results of fluorescence staining and the inhibitory effect on the growth of HCC SUN475 showed that isoliquiritigenin induced the apoptosis cells increased gradually. The interaction of ISO with DNA can cause the denaturation temperature to be increased, which indicated that the stabilization of the DNA helix was increased in the presence of ISO, which indicated that the results provide strong evidence for intercalative binding of ISO with DNA.

Deubiquitinating enzyme Josephin-2 stabilizes PHGDH to promote a cancer stem cell phenotype in hepatocellular carcinoma.

BACKGROUND: Deubiquitinating enzymes (DUBs) have been shown to be possible targets for hepatocellular carcinoma (HCC) treatment. OBJECTIVE: This study was designed to reveal the effect and underlying mechanism of Josephin-2, a relatively newly defined DUB, in HCC progression. METHODS: SNU-387 and PLC/PRF/5 cells were used for in vitro functional assays. The levels of Josephin-2 and phosphoglycerate dehydrogenase (PHGDH) were determined using RT-qPCR and western blotting. Cell proliferation, migration and invasion were assessed by CCK-8, colony formation and Transwell. Spheroid-forming assay was performed to assess the cancer stem cell (CSC)-phenotype of HCC cells. A xenograft mice model was applied to verify the effect of Josephin-2 on HCC cell growth in vivo. RESULTS: Herein, we showed that Josephin-2 expression was negatively correlated with HCC patient survival in data from the online database. Cell experiments indicated that knockdown of Josephin-2 attenuated HCC cell malignant biological behaviors. Besides, Josephin-2 silencing also decreased the spheroid-formation while inhibited the expression of CSC biomarkers (CD133, OCT4, SOX2 and EpCAM) in HCC cells. Mechanistically, Josephin-2 had a deubiquitinating activity towards the regulation of PHGDH protein, the rate-limiting enzyme in the first step of serine biosynthesis pathway. Depletion of Josephin-2 enhanced the ubiquitination degradation of PHGDH and ultimately inhibited the proliferation and CSC-phenotype of HCC in vitro and in vivo. CONCLUSION: Our work uncovered the regulatory effects of Josephin-2 on PHGDH protein stability and profiled its contribution in HCC malignant progression, which might provide a potential therapeutic target for HCC.

Population dynamics of Phaius flavus in southeast China: Reproductive strategies and plants conservation.

Because of species diversity and troubling conservation status in the wild, Orchidaceae has been one of the taxa with most concern in population ecological research for a long time. Although Orchidaceae is a group with high adaptability, they have become endangered for complex and various reasons such as the germination? difficulty and habitat loss, which makes it difficult to develop an accurate protection strategy. Phaius flavus is a terrestrial orchid which used to be widely distributed in central and southern Asia; however, large populations are difficult to find in the wild. Thus, the aim of this study was to provide a new perspective for conserving endangered P. flavus by investigating the mechanisms of its population decline; we established time-specific life and fertility tables, age pyramids, survival curves, and mortality curves for this plant and then conducted Leslie matrix model. We found that both of the populations from Wuhu Mount (WM) and Luohan Mount (LM) showed declining trends and exhibited pot-shaped age pyramids, low net reproductive rates, and negative intrinsic growth rates. The population from the Beikengding Mount (BM) showed a stable status with a bell-shaped age pyramid. However, it has a significant risk of decline because of the low net reproductive rate and intrinsic growth rate. This study use time-specific life and fertility tables, age pyramids, survival curves, and mortality curves, showed that the population decline of P. flavus could be attributed to 1) the shortage of seedlings caused by the low germination rate in the wild and 2) the loss of adult individuals caused by anthropogenic disturbances. To protect this species from extinction in these areas, we suggest that human activities in these habitats should be strictly forbidden and ex situ conservation of this plant in botanical gardens is also necessary.

A gigawatt level repetitive rate adjustable magnetic pulse compressor.

In this paper, a gigawatt level repetitive rate adjustable magnetic pulse compressor is investigated both numerically and experimentally. The device has advantages of high power level, high repetitive rate achievability, and long lifetime reliability. Importantly, dominate parameters including the saturation time, the peak voltage, and even the compression ratio can be potentially adjusted continuously and reliably, which significantly expands the applicable area of the device and generators based on it. Specifically, a two-stage adjustable magnetic pulse compressor, utilized for charging the pulse forming network of a high power pulse generator, is designed with different compression ratios of 25 and 18 through an optimized design process. Equivalent circuit analysis shows that the modification of compression ratio can be achieved by just changing the turn number of the winding. At the same time, increasing inductance of the grounded inductor will decrease the peak voltage and delay the charging process. Based on these analyses, an adjustable compressor was built and studied experimentally in both the single shot mode and repetitive rate mode. Pulses with peak voltage of 60 kV and energy per pulse of 360 J were obtained in the experiment. The rise times of the pulses were compressed from 25 µs to 1 µs and from 18 µs to 1 µs, respectively, at repetitive rate of 20 Hz with good repeatability. Experimental results show reasonable agreement with analyses.

Injuries of myocardial cells and changes of myocardial enzymes after firearm wound-induced intestinal perforation in porcine abdomen.

This study aims to observe the changes of myocardial injuries after the firearm wound-induced intestinal perforation in porcine abdomen. 42 healthy Landrace piglets were randomly divided into the control group and the injury group, which was then subdivided into the post-injury 1 h, 2 h, 4 h, 8 h, 12 h and 24 h subgroup. the LDH, CK and CK-MB levels of each group, as well as the plasma endotoxin, were determined and compared. The plasma endotoxin levels of the experimental groups were significantly higher than the control group, and the light microscope observation revealed that the 8 h, 12 h and 24 h subgroup appeared the gradually-aggravated myocardial cell edema and degeneration; the electron microscope revealed that the 4 h, 8 h, 12 h and 24 h subgroup appeared the mitochondrial swelling and dissolution gradually; the serum levels of LDH, CK and CK-MB of each experimental group were higher than the control group. The abdominal firearm wound-induced intestinal perforation would lead to the damaged changes of myocardial morphology and enzymes, which would aggravate as time went along.