Search

Aim: To understand how liver sinusoidal endothelial cells (LSECs) respond to nonalcoholic steatohepatitis (NASH). Methods: We profiled single-LSEC from livers of control and MCD-fed mice. The functions of C-Kit+-LSECs were determined using coculture and bone marrow transplantation (BMT) methods. Results: Three special clusters of single-LSEC were differentiated. C-Kit+-LSECs of cluster 0, Msr1+-LSECs of cluster 1 and Bmp4+Selp+-VECs of cluster 2 were revealed, and these cells with diverse ectopic expressions of genes participated in regulation of endothelial, fibrosis and lipid metabolism in NASH. The number of C-Kit+-primary LSECs isolated from MCD mice was lower than control mice. Immunofluorescence co-staining of CD31 and C-KIT showed C-Kit+-LSECs located in hepatic sinusoid were also reduced in NASH patients and MCD mice, compared to AIH patients and control mice respectively. Interestingly, lipotoxic hepatocytes/HSCs cocultured with C-Kit+-LSECs or the livers of MCD mice receipting of C-Kit+-BMCs (bone marrow cells) showed less steatosis, inflammation and fibrosis, higher expression of prolipolytic FXR and PPAR-α, lower expression of TNF-α and α-SMA. Furthermore, coculturing or BMT of C-Kit+-endothelial derived cells could increase the levels of hepatic mitochondrial LC3B, decrease the degree of mitochondrial damage and ROS production through activating Pink1-mediated mitophagy pathway in NASH. Conclusions: Hence, a novel transcriptomic view of LSECs was revealed to have heterogeneity and complexity in NASH. Importantly, a cluster of C-Kit+-LSECs was confirmed to recovery Pink1-related mitophagy and NASH progression. [ABSTRACT FROM AUTHOR]

Background: The role of macrophages in the pathogenesis of nonalcoholic steatohepatitis (NASH) is complex and unclear. Methods: Single-cell RNA sequencing was performed on nonparenchymal cells isolated from NASH and control mice. The expression of Vsig4+ macrophages was verified by qPCR, flow cytometry and immunohistochemistry. Primary hepatic macrophages were cocultured with primary hepatocytes or hepatic stellate cells (LX2) cells by Transwell to detect immunofluorescence and oil red O staining. Results: Two main single macrophage subsets were identified that exhibited a significant change in cell percentage when NASH occurred: resident Kupffer cells (KCs; Cluster 2) and lipid-associated macrophages (LAMs; Cluster 13). Nearly 82% of resident single KCs in Cluster 2 specifically expressed Cd163, and an inhibited subgroup of Cd163+ resident single-KCs was suggested to be protective against NASH. Similar to Cd163, Vsig4 was both enriched in and specific to Cluster 2. The percentage of Vsig4+-KCs was significantly decreased in NASH in vivo and in vitro. Hepatocytes and hepatic stellate cells produced less lipid droplet accumulation, proinflammatory protein (TNF-α) and profibrotic protein (α-SMA) in response to coculture with Vsig4+-KCs than in those cocultured with lipotoxic KCs. Conclusions: A subgroup of Vsig4+ resident single-KCs was shown to improve hepatic inflammation and fibrosis in NASH. [ABSTRACT FROM AUTHOR]

Plant communities that colonize high-elevation zones generally have short growing seasons which lead to specialized adaptations in such zones. In montane areas of Taiwan, Yushan bamboo (YB, Yushania niitakayamensis ) is dominant at elevations ranging from 2500 to 3300 m and grows in a grassland-like open habitat. In this study, hexapods were collected from YB bimonthly between 2009 and 2012 by using a sweeping net. The composition of and several bioindices for the hexapods were determined, and multivariate analyses were conducted to explore the dynamics and seasonal distribution of the hexapods. A total of 32,000 individuals belonging to 11 orders and 113 families were collected, with adult individuals being collected more frequently in warmer seasons (from June to October). Of the sampled individuals, 90% belonged to the orders Collembola (42%), Hemiptera (35%), and Hymenoptera (13%). The number of individuals belonging to Hemiptera were stable in all seasons, and the number of hymenopteran wasps was influenced by temperature and exhibited a stable dynamic pattern. The number of individuals belonging to Collembola fluctuated dramatically. The multivariate analyses revealed that the collected hexapods could be divided into two major family groups according to survey season ( i.e. , summer and winter groups). Several families were collected only in summer, but a few were collected only in winter. Eigenvalues obtained from a principal component analysis revealed that the families Chironomidae, Delphacidae, Entomobryidae, Hypogastruridae, Sminthuridae, and Thripidae (all dominant) were the major contributors to the winter group. These families were abundant all year, although some were more abundant during winter. The three dominant orders Collembola, Hemiptera, and Hymenoptera, each of which has a distinct community structure and dynamic pattern, may have their own adaptive mechanisms in the subtropical regions of Taiwan. Hemiptera individuals, which feed on YB, were most abundant in the adult stage in summer and in the nymphal stage in winter. The abundance of parasitic hymenopteran wasps, which had stable dynamic patterns, was associated with that of their host insects and temperature. The drastic fluctuations in the abundance of Collembola may have been caused by abiotic factors, such as precipitation and microhabitat factors. The early onset of spring and the late onset of winter might also affect the dynamics of the studied hexapods.

An HPLC-UV method has been developed for the determination of valibose, miglitol, voglibose and acarbose, the four anti-diabetic drugs. The separation was accomplished successfully by using reversed phase chromatography (Prevail carbohydrate column, 250 mm x 4.6 mm, 5 microm) with a gradient acetonitrile-phosphate buffer solution (pH 8.0) at a wavelength of 210 nm. Furthermore, the method of a high-performance liquid chromatography coupled with ESI-MS in positive ionization mode has been established. These two methods were successfully applied to the assay and qualitative detection of four alpha-glucosidase inhibitors in the potential counterfeit anti-diabetic drugs.