Network analysis and experimental verification of Salvia miltiorrhiza Bunge-Reynoutria japonica Houtt. drug pair in the treatment of non-alcoholic fatty liver disease.

CONTEXT: There are currently no approved specific clinical drugs for non-alcoholic fatty liver disease (NAFLD). Salvia miltiorrhiza Bunge-Reynoutria japonica Houtt. drug pair (SRDP) has been widely used in the treatment of chronic liver diseases. However, the mechanism of SRDP treating NAFLD remains unclear. OBJECTIVE: Based on network analysis and in vitro experimental verification, we investigated the effect of SRDP on lipid deposition and explored its possible mechanism for the treatment of NAFLD. METHODS: The TCMSP platform was used to screen the active metabolites of SRDP and corresponding targets. The GeneCards and OMIM databases were used to screen the NAFLD targets. The drug-disease intersecting targets were extracted to obtain the potential targets. Then the protein-protein interaction (PPI) and drug-active metabolites-target-disease network map was constructed. The DAVID database was performed to GO and KEGG pathway enrichment analysis for the intersecting targets. The core active metabolite and signaling pathway were verified by in vitro experiments. RESULTS: Network analysis predicted 59 active metabolites and 89 targets of SRDP for the treatment of NAFLD. 112 signaling pathways were enriched for KEGG pathways, including PI3K-AKT signaling pathway,etc. It was confirmed that luteolin, the core active metabolite of SRDP, effectively reduced fat accumulation and intracellular triglyceride content in HepG2 fatty liver cell model. Luteolin could inhibit mTOR pathway by inhibiting PI3K-AKT signaling pathway phosphorylation, thereby activating autophagy to alleviate NAFLD. DISCUSSION AND CONCLUSION: The results of this study validate and predict the possible role of various active metabolites of SRDP in the treatment of NAFLD through multiple targets and signaling pathways. The core active metabolite of SRDP, luteolin can alleviate NAFLD by acting on the PI3K-AKT-mTOR signaling pathway to induce autophagy.

Neotype designation and redescription of Tropidophorus guangxiensis Wen, 1992 (Squamata: Sauria: Scincidae), with description of a new subspecies from central South China.

The genus Tropidophorus consists of small semi-aquatic lizards that dwell in lowland forest steams (Barbour, 1921; Bauer & Jackman, 2008). Here, we designate the neotype and re-describe T. guangxiensis Wen, 1992 based on newly collected topotypic specimens. We also describe a new subspecies from Xuefeng Mountain, Hongjiang County, Hunan Province, central South China. Based on two mitochondrial genes (12S rRNA and 16S rRNA), the phylogenetic position of T. guangxiensis is allocated for the first time. Additionally, our data strongly support that the new subspecies is phylogenetically closely related to T. g. guangxiensis. We also present a morphological identification key for known species and subspecies of Tropidophorus in China.

A new species of the genus Megophrys (Amphibia: Anura: Megophryidae) from Hunan, China.

We describe a new species of Megophryidae frog, Megophrys shunhuangensis sp. nov., from Hunan Province, Southern China. It can be distinguished from other known species in the subgenus Panophrys by morphological characters, bioacoustic data and a molecular divergence in the mitochondrial 16S rRNA gene similar to that found among other species of Panophrys. M. shunhuangensis sp. nov. is characterized by a relatively small body size, with adult females measuring 37.6 mm and adult males measuring 30.3-33.6 mm in snout to vent length; maxillary teeth present, vomerine teeth absent; tongue smooth, not notched behind; hindlimb slender, heels overlapping, tibio-tarsal articulation reaches forward between the nasal and tip of snout. Molecular phylogenetic analyses also show that M. sp. 6 from (Mao'er Shan, China) from Chen et al. (2017) and M. sp. 24 (Anjiangping and Mao'er Shan, China) from Liu et al. (2018) may be the same species as M. shunhuangensis sp. nov., we consequently speculate that the new species also distributed in Anjiangping and Mao'er Shan, China. At present, the genus Megophrys contains 85 species of which 48 species are distributed in China, and 30 belong to the subgenus Panophrys.

Rhinogobius maculagenys, A new species of freshwater goby (Teleostei: Gobiidae) from Hunan, China.

A new freshwater goby, Rhinogobius maculagenys sp. nov., was collected from Hunan Province in Southern China. This species can be distinguished from all congeners by a combination of the following features: first dorsal fin with 6 spines; second dorsal fin with a single spine and 7-9 segmented rays; anal fin with a single spine and 6-8 segmented rays; pectoral fin with 16 segmented rays; 32-34 longitudinal scales; 9-13 transverse scales; 11+16=27 vertebrae; pore ω1 missing; head and body yellowish brown; cheek and opercle yellowish brown with over 30 small orange spots, branchiostegal membrane yellow with over 10 small orange spots in males and white and spotless in females; first dorsal fin trapezoidal in males and nearly semicircular in females, with large bright blue blotch in front of second spine; spines 4 and 5 longest, rear tip extending to base of second branched ray of second dorsal fin in males when adpressed, but just reaching or not reaching anterior margin of second dorsal fin in females; caudal fin with 5-6 vertical rows of brown spots; flank with several longitudinal rows of blackish-brown spots; and belly pale white.

Molecular cloning, characterization and expression of FSH and LH beta subunits from grass carp (Ctenopharyngodon idella).

Follicle-stimulating hormone beta subunit (FSHbeta) and luteinizing hormone beta subunit (LHbeta) have been cloned and characterized from the pituitary of grass carp (Ctenopharyngodon idella). The full length of FSHbeta and LHbeta cDNA was 393 bp and 441 bp, with open reading frame encoding proteins of 130 and 146 amino acids, respectively. Phylogenetic analysis of the proteins of FSHbeta and LHbeta showed a high homology with other fishes. Homology analysis also indicated that LHbeta has higher conservation than FSHbeta. The expression analysis of grass carp FSHbeta and LHbeta by RT-PCR suggested that they were only expressed in the pituitary. Real-time quantitative PCR protocols were developed and validated to measure FSHbeta and LHbeta mRNAs during ovarian development. The FSHbeta and LHbeta mRNA level was very low in the pituitaries of early-pubertal fish and significantly increased during the ovulation period. These results suggested that in grass carp the gonadotropins synthesized synchronously in order for asynchronous oogenesis to take place.