[Intervention effect of Erchen Decoction on methionine and choline deficient diet-induced non-alcoholic steatohepatitis in mice].

This study investigated the effect of Erchen Decoction(ECD) on the prevention of non-alcoholic steatohepatitis(NASH) in mice and explored its possible mechanism, so as to provide scientific data for the clinical application of ECD in the prevention of NASH. C57BL/6 male mice were randomly divided into normal group(methionine and choline supplement, MCS), model group(methionine and choline deficient, MCD), low-dose ECD group(ECD＿L, 6 g·kg~(-1)), medium-dose ECD group(ECD＿M, 12 g·kg~(-1)), and high-dose ECD group(ECD＿H, 24 g·kg~(-1)), with eight mice in each group. The MCS group was fed with an MCS diet, and the other groups were fed with an MCD diet. The mice in each group were given corresponding diets, but the drug intervention group was given low-, medium-, and high-dose ECD(10 mL·kg~(-1)·d~(-1)) by intragastric administration for six weeks on the basis of MCD diet feeding, and the mice could eat and drink freely during the whole experiment. At the end of the experiment, mice were fasted overnight(12 h) and were anesthetized with 20% urethane. Thereafter, the blood and liver tissue were collected. The serum was used to detect the levels of alanine aminotransferase(ALT), aspartate aminotransaminase(AST), interleukin-1ß(IL-1ß), interleukin-6(IL-6), interleukin-10(IL-10), and tumor necrosis factor-α(TNF-α). Liver tissue was processed by hematoxylin-eosin(HE) staining and used for hepatic histological analysis and detection of the expression levels of genes and proteins related to nuclear factor erythroid 2-related factor 2/glutathione peroxidase 4(Nrf2/GPX4) pathway by real-time quantitative reverse transcriptase-polymerase chain reaction(RT-qPCR) and Western blot analysis, respectively. The results showed that compared with the MCS group, the MCD group showed higher serum ALT and AST levels; the HE staining exhibited fat vacuoles and obvious inflammatory cell infiltration in liver tissue; serum IL-1ß, IL-6, and TNF-α levels were significantly increased, and the serum IL-10 level was significantly decreased. The mRNA expressions of fatty acid synthase(FASN), monocyte chemoattractant protein-1(MCP-1), and IL-1ß in liver tissue were significantly up-regulated, while those of GPX4, Nrf2, and NAD(P)H:quinine oxidoreductase(NQO1) were significantly down-regulated. Compared with the MCD group, the serum ALT and AST levels of ECD＿M and ECD＿H groups were significantly decreased, and the AST level in the ECD＿L group was significantly decreased. The number of fat vacuoles and the degree of inflammatory cell infiltration in liver tissue were improved; serum IL-1ß, IL-6, and TNF-α levels were significantly decreased, but the serum IL-10 level was significantly increased only in the ECD＿H group. The mRNA expressions of FASN, MCP-1, and IL-1ß in liver tissue were significantly down-regulated, and those of GPX4 and NQO1 were significantly up-regulated. The mRNA expressions of Nrf2 in ECD＿M and ECD＿H groups were significantly up-regulated. Western blot results showed that compared with the MCD group, the protein expression levels of Nrf2 and GPX4 in each group were significantly increased after ECD administration, and the protein expression level of FASN was significantly decreased; the protein expression of NQO1 was increased in ECD＿M and ECD＿H groups. In summary, ECD can reduce hepatic lipid accumulation, oxidative stress, liver inflammation, and liver injury in NASH mice, which may be related to the activation of the Nrf2/GPX4 pathway.

Multi-gene phylogenetic evidence indicates that Pleurodesmospora belongs in Cordycipitaceae (Hypocreales, Hypocreomycetidae) and Pleurodesmospora lepidopterorum sp. nov. on pupa from China.

A new species, Pleurodesmospora lepidopterorum, isolated from a pupa, is introduced. Morphological comparisons and phylogenetic analyses based on multigene datasets (ITS+RPB1+RPB2+TEF) support the establishment of the new species. Pleurodesmospora lepidopterorum is distinguished from P. coccorum by its longer conidiogenous pegs located in the terminal or lateral conidiophores, and smaller subglobose or ellipsoidal conidia. A combined dataset of RPB1, RPB2, and TEF confirmed the taxonomic placement of Pleurodesmospora in Cordycipitaceae for the first time.

Morphological and phylogenetic characterisations reveal three new species of Samsoniella (Cordycipitaceae, Hypocreales) from Guizhou, China.

Samsoniella species have been found on lepidopteran larvae or pupae buried in soil or leaf litter. Three new species, Samsoniella hymenopterorum, S. coleopterorum and S. lepidopterorum, parasitic on hymenopteran larvae, coleopteran larvae and lepidopteran pupae, respectively, are reported. Morphological comparisons with extant species and DNA-based phylogenies from analysis of a multigene (ITS, RPB1, RPB2 and TEF) dataset supported the establishment of the new species. Unusually, all three new species have mononematous conidiophores. The new species are clearly distinct from other species in Samsoniella occurring in separate subclades.

Characterization of Nine Compounds Isolated from the Acid Hydrolysate of Lonicera fulvotomentosa Hsu et S. C. Cheng and Evaluation of Their In Vitro Activity towards HIV Protease.

In this study, we isolated nine compounds from the acid hydrolysate of the flower buds of Lonicera fulvotomentosa Hsu et S. C. Cheng and characterized their chemical structures using 1H-NMR, 13C-NMR, and electron ionization mass spectroscopy (EI-MS). These compounds were identified as ß-sitosterol (1), 5,5'-dibutoxy-2,2'-bifuran (2), nonacosane-10-ol (3), ethyl (3ß)-3,23-dihydroxyolean-12-en-28-oate (4), oleanolic acid (5), ethyl caffeate (6), caffeic acid (7), isovanillin (8), and hederagenin (9), with 4 as a new triterpene compound. Inhibitory activity against human immunodeficiency virus (HIV) protease was also evaluated for the compounds, and only ethyl caffeate, caffeic acid, and isovanillin (6, 7, and 8) exhibited inhibitory effects, with IC50 values of 1.0 µM, 1.5 µM, and 3.5 µM, respectively. Molecular docking with energy minimization and subsequent molecular dynamic (MD) simulation showed that ethyl caffeate and caffeic acid bound to the active site of HIV protease, while isovanillin drifted out from the active site and dissociated into bulk water during MD simulations, and most of the binding residues of HIV protease have been previously identified for HIV protease inhibitors. These results suggest that caffeic acid derivatives may possess inhibitory activities towards HIV protease other than previously reported inhibitory activities against HIV integrase, and thus ethyl caffeate and caffeic acid could be used as lead compounds in developing potential HIV protease inhibitors, and possibly even dual-function inhibitors against HIV.

Three novel insect-associated species of Simplicillium (Cordycipitaceae, Hypocreales) from Southwest China.

In this paper, we introduce three new species of Simplicillium, viz. S. cicadellidae, S. formicidae and S. lepidopterorum, which were isolated from an infected leafhopper, ant and carpenterworm, respectively. Morphological comparisons and phylogenetic analyses based on multigene datasets (LSU+RPB1+RPB2+TEF and ITS+LSU) support the establishment of the three new species. Simplicillium cicadellidae was distinguished from other species in morphological characteristics by having smaller phialides and ellipsoidal conidia, and lacking octahedral crystals. The reverse of colonies were yellowish (#FFBF00), especially in the middle, and radially sulcate. Simplicillium formicidae was morphologically distinguished from other by having longer phialides and filiform to fusoid conidia, and by lacking octahedral crystals. Simplicillium lepidopterorum was morphologically distinguished from other species by having smaller, ellipsoidal to fusiform conidia, and by lacking octahedral crystals. The reverse of the colony was pale white. The three new species are likely to be nourished by plant to animal (especially insect) nutrients based on the evolutionary pattern of the Hypocreales, and they are described herein as being clearly distinct from other species in Simplicillium.

[Construction of expression vector of ALAg and immune protection of its recombinant protein induced in mice].

OBJECTIVE: To determine the candidate genes for engineering vaccine of Ascaris lumbricoides. METHODS: pMD18-T-ALAg and plasmid expression vector pET-28a(+) were digested with BamH I and EcoR I and linked to each other. The resultant plasmid pET-28a(+)-ALAg was transferred into E. coli BL21 (DE3) and its expression was induced with IPTG, and the recombinant ALAg(rALAg) was purified. A total of 30 mice were equally divided into 3 groups, the mice in each group were injected with rALAg-FCA, FCA and PBS respectively, then they were attacked by infectious eggs of Ascaris (3 600 per mouse). The IgG levels in sera of mice in each group were detected by indirect ELASA. RESULTS: rALAg was recognized by the sera from repeatedly Ascaris lumbricoides inoculated rabbits. The numbers of larvae of Ascaris lumbricoides from liver and lung of mice were 25.30 +/- 4.55 in the rALAg-FCA group and 57.60 +/- 5.76 in the PBS group, respectively, the former being the reducing rate of 69.26%, and the difference among the 3 groups showed statistical significances (P < 0.01). The IgG levels (A450 value) of the rALAg-FCA, FCA and PBS groups were 0.858 +/- 0.003, 0.149 +/- 0.004 and 0.134 +/- 0.004, respectively, there were statistical differences among them (P < 0.01). CONCLUSION: ALAg can be used as a candidate gene of genetic engineering vaccine of Ascaris lumbricoides.

Evaluation of the colonization capabilities of Salmonella Enteritidis in quails using an RT-PCR approach.

We used a real-time PCR assay and indirect fluorescent antibody (IFA) assay to detect genomic DNA of Salmonella Enteritidis in the internal organs of quails after an oral challenge. The results showed that S. Enteritidis was detected in all the samples at different time points. This study will assist a future understanding of the pathogenesis of S. Enteritidis.

Evaluation of the efficacy of a recombinant Entamoeba histolytica cysteine proteinase gene (EhCP5) antigen in Minipig.

Entamoeba histolytica cysteine proteinase gene 5(EhCP5) is one of the major proteinase genes of all EhCP-transcripts. The amebiasis cysteine proteinase gene encoding an antigen from E. histolytica, as well as the recombinant EhCP5, obtained by cloning and expression of the EhCP5 gene in heterologous host Escherichia coli BL-21 (DE3), were used to evaluate their ability to induce immune protective responses in Minipig against challenge infection in a minipig-E. histolytica model. There was a 52.27% reduction (P<0.001) in the group of recovery of challenged E. histolytica compared with that in the control group. Specific anti-EhCP5 antibodies from immune protected minipig had significantly higher levels of immunoglobulin G (IgG) (P<0.0001). Our data will help to know the mechanism of vaccinal protection of E. histolytica.

The population of a high-virulence strain of Salmonella enterica serovar Enteritidis in subcutaneously infected partridge: a quantitative time-course study using real-time PCR.

This research was undertaken to determine the population of a high-virulence strain of Salmonella enterica serovar Enteritidis in partridge by a fluorescent quencher PCR assay and to correlate these findings with the results obtained from the immunohistochemical localization and histopathological examinations of selected Salmonella enterica serovar Enteritidis-infected tissues. To make the results meaningful, a side-by-side bacteriology method (indirect immuno-fluorescent antibody staining) was performed too. The results of indirect immuno-fluorescent antibody staining and immunohistochemical localization were similar to the fluorescent quencher PCR assay. The time course of the appearance of bacterial antigens and tissue lesions in various tissues was coincident with the levels of the bacterial DNA loads at the infection sites. This suggests that Salmonella enterica serovar Enteritidis loads in internal organs are closely correlated with the progression of the infection.

[Determination of glycyrrhizic acid in different decoctions of sanaotang by HPLC and comparison with antifugal effects in vitro].

OBJECTIVE: To investigate the contents of glycyrrhizic acid in hejian decoction (mixed the traditional Chinese herbs together, then boiling them with water) and the fenjian decoction (boiling the single traditional Chinese herb with water separately, then mixed the abstracts) of Sanaotang (composed of Ephedra sinica, Prunus armeniaca and Glycyrrhiza uralensis) and to compare with their anti-bacterial activities in vitro. METHOD: A HPLC method was established with a Agilent Zorbax SB-C18 column (4. 6 mm x 250 mm, 5 microm), a mobile phase of acetonitrile-0.2% acetic acid solution (35:65), a flow rate of 1 mL x min(-1) and a detection wavelength of 254 nmn in order to determine the contents of glycyrrhizic acid minimal bacterial inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) antagonized the common bacteria in different decoctions were rieasured in vitro by employing dilution method. RESULT: The average content of glycyrrhizic acid of the hejian decoction was higher than that of the fenjian decoction. The hejian decoction could display the inhibitory bactericidal activity to Aeruginosus bacillus, but the fenjian decoction could not. And to Staphylococcus aureus, the inhibitory bactericidal activity the hejian decoction was slightly stronger than that of the fenjian decoction. CONCLUSION: Comparing with that of the fenjian decoction, the content of glycyrrhizic acid of the hejian decoction was higher and the anti-bacterial activities was stronger.