Analysis of microbial diversity in the midgut of Haemaphysalis longicornis infected with severe fever with thrombocytopenia syndrome virus / 中国热带医学

@#Abstract: Objective　To investigate the composition and diversity of midgut microbial community of Haemaphysalis longicornis infected with severe fever with thrombocytopenia syndrome virus (SFTSV)． Methods　The midgut DNA of three group Haemaphysalis longicornis infected with SFTSV was extracted, and the 16S rDNA gene of the sample was sequenced by HiSeq platform. The composition and diversity of endosymbiotic microbial community were clarified by OTU cluster analysis and alpha diversity analysis. Results　The midgut microbial clusters of the three groups infected with SFTSV were 143, 113, 163 OTUs respectively; the sparsity curve and abundance grade curve showed that the data had sufficient sequencing depth, and the midgut of Haemaphysalis longicornis infected with SFTSV was rich in microbial composition, but the species distribution was uneven. The analysis of microbial community composition showed that Proteobacteria, Firmicutes and Actinobacteria were the main dominant bacteria at the phyla level. At the class level, Gammaproteobacteria, Bacilli, Betaproteobacteria and Actinomycetia were the main dominant bacteria. At the order level, Legionellales, Bacillales, Burkholderiales and Actinomycetales were the main dominant orders. At the family level, Coxiellaceae, Bacillaceae, Moraxellaceae and Rhodococcaceae were the main dominant families. At the genus level, the relative abundance of Coxiella was the highest, followed by Aeribaillus and Azonexus. Alpha diversity analysis showed that the average Shannon index was 139.67, the average Simpson index was 0.48, the average Chao index was 145.06, and the average ACE index was 147.11. Conclusions　The species diversity of intestinal microorganisms in Haemaphysalis longicornis infected with SFTSV is rich. The results provide a basis for further exploring the interaction between intestinal microbes of Haemaphysalis longicornis and SFTSV and developing new ideas for the prevention and control of ticks and tick-borne diseases.

Design, synthesis and activity evaluation of protein arginine methyltransferase 5 inhibitor / 药学学报

Protein arginine methyltransferase 5 (PRMT5) is an important type II human methyltransferase. It catalyzes the symmetrical double-methylation of many histones and non-histones, and it is highly expressed in many kinds of tumors. PRMT5 has been proven to be a potential new target for cancer treatment. Based on the reported crystal complex of EPZ015666 with PRMT5, a series of new compounds was designed using GSK3326595 (EPZ015938) as the lead compound and using the conformational restriction approach. We found that compounds B8 and the C series of derivatives displayed enzyme inhibitory activity comparable to that of GSK3326595. Compounds C3 and C4 showed poor permeability in Caco-2 cells, and that might be one of the reasons for their poor anti-proliferative activity against Z-138 cells. These data provide insights for further structural optimization.

Effect of Yanghetang on Apoptosis of Triple Negative Breast Cancer Cell Line MDA-MB-231 Based on Egr1/p21 Signaling Pathway / 中国实验方剂学杂志

Objective::To investigate the effect of ancient recipe Yanghetang and its drug-contained serum on apoptosis of triple negative breast cancer cell line MDA-MB-231 based on the signal pathway of early growth response gene 1 (Egr1)/cyclin dependent inhibitor (p21). Method::The drug-containing serum of Yanghetang was prepared from rats, and MDA-MB-231 cells were cultured in vitro. The blank control group was set, and MDA-MB-231 cells in Yanghetang high, middle, and low dose groups (23.4, 11.7, 5.85 g·kg-1) were intervened for 24, 48, 72 h, respectively. After that, the cell counting kit-8(CCK-8) method was used to detect the cell proliferation of each group. The blank control group was set, while MDA-MB-231 cells in Yanghetang high, middle, and low dose groups (23.4, 11.7, 5.85 g·kg-1) were treated for 48 h, and then flow cytometry was used to detect the apoptosis of each group and the distribution of cell cycle. The expression of Egr1 and p21 mRNA in each group was detected by quantificational Real-time polymerase chain reaction (Real-time PCR), while the expression of Egr1 and p21 protein in each group was detected by Western blot. Result::After MDA-MB-231 cells were intervened by Yanghetang for 24, 48, 72 h, MDA-MB-231 cell proliferation was significantly inhibited in Yanghetang high and middle dose groups as compared with the blank control group (P<0.01). After MDA-MB-231 cells were intervened by Yanghetang for 48 h, the apoptosis was significantly increased in Yanghetang high and middle dose groups as compared with the blank control group (P<0.05, P<0.01). In the Yanghetang high, middle dose groups, the proportion of cell cycle G0/G1 phase decreased, and the proportion of G2/M phase increased (P<0.05, P<0.01). The mRNA and protein expressions of Egr1, p21 were increased in Yanghetang high and middle groups (P<0.05, P<0.01). Conclusion::Yanghetang can activate Egr1/p21 signaling pathway in MDA-MB-231 cells, increase the expression of Egr1 and p21, and cause G2/M cell cycle arrest, thereby inducing apoptosis of MDA-MB-231 cells.

Effect of Serum Containing Yanghetang on Apoptosis of MCF-7 Cells in Breast Cancer Through p38/STAT3 Signaling Pathway / 中国实验方剂学杂志

Objective::To investigate the effect of serum containing Yanghetang (YHT) on the apoptosis of MCF-7 cells in breast cancer based on the mitogen-activated protein kinase (p38)/signal transduction and transcriptional activator 3 (STAT3) signal pathway. Method::YHT liquid with crude drug 1 g·mL-1 was prepared. Female SD rats were randomly divided into control group (distilled water), and high, medium and low-dose YHT groups (24, 12, 6 g·kg-1). YHT-medicated serum was prepared, and 10%medicated serum was used to intervene MCF-7 cells. Cell proliferation and cytotoxicity assay (CCK-8) was used to detect the effect of serum containing YHT on MCF-7 cell proliferation, apoptosis of MCF-7 cells was detected by flow cytometry protein expressions of p38 and STAT3 were detected by Western blot, Quantitative Real-time PCR (Real-time PCR) was used to detect the expressions of B-cell lymphoma/leukemia-xl(Bcl-xl) and Survivin mRNA. Result::CCK-8 assay showed that YHT serum inhibited the proliferation of MCF-7 cells in a time and dose-dependent manner compared with the blank group. The inhibitory effect was most obvious in the high-dose group, with the inhibition rates of 38%, 45%and 54%at different time points (P<0.01). Flow cytometry showed that, compared with the blank group, the apoptosis rate in the medium and high-dose groups increased significantly in a dose-dependent manner, with the apoptosis rates at 11.6%and 16.5%respectively (P<0.05, P<0.01). Western blot analysis showed that, compared with the blank group, the expressions of p38 and STAT3 protein was decreased in high, medium-dose YHT groups (P<0.01) in a dose-dependent manner. Compared with the blank group, the expressions of Bcl-xl and Survivin mRNA were decreased in high, medium-dose YHT groups (P<0.05, P<0.01) in a dose-dependent manner. Conclusion::YHT serum can promote the apoptosis of MCF-7 cells in breast cancer, which may be related to the p38/ STAT3 signaling pathway.

Synovial inflammation of a rabbit model of early-stage knee osteoarthritis induced by low concentration of papain at different time points / 中国组织工程研究

BACKGROUND:Synovitis plays an important role in the occurrence and development of early knee osteoarthritis.OBJECTIVE:To compare synovial inflammation in a rabbit mode of knee osteoarthritis induced by injecting low concentration of papain at different time points,thus providing reference for the study on synovitis in knee osteoarthritis.METHODS:Twenty-four New Zealand white rabbits were divided randomly into four groups,and received the injection of 0.5 mL mixture of 2％ papain with 0.03 mol/L L-cysteine into the right knee at 1,4 and 7 days,respectively.The model rabbits were respectively sacrificed at 1,2 and 3 weeks after the last injection,and the rabbits in the blank control group were killed at 3 weeks.The local skin temperature and circumference of the knee were recorded,and the synovium and infrapatellar fat pad were separated from the right knees for histopathological observation and ELSA.RESULTS AND CONCLUSION:At the 1stweek after modeling,joint effusion was significantly increased,local skin temperature and circumference of knee joint were higher than those at the 2nd,and 3rd weeks.The levels of interleukin-1,tumor necrosis factor-α and matrix metalloproteinase-13 in the synovium in the three experimental groups were higher than those in the blank control group at 1,2 and 3 weeks after modeling;the levels peaked in the 1st week,but no significant fluctuation appeared in the 2nd and 3rd weeks.There were synovial tissue hyperplasia,thickening,and inflammatory cell infiltration in the 1st,2nd and 3rd weeks,and the proliferation of synovial tissue increased significantly with time.These findings indicate that the intra-articular injection of low concentration of papain and 0.03 mol/L L-cysteine mixtures contributes to a rabbit model of knee synovial inflammation within 1 week,with significantly joint effusion.However,significant synovial tissue thickening and vascular hyperplasia are observed;meanwhile,the joint effusion is decreased obviously.

Effect of solution environments on ceramic membrane microfiltration of model system of Chinese medicines / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the effect of differents solution environments on the ceramic membrane microfiltration of model system of Chinese medicines.</p><p><b>METHOD</b>Taking binary system of soybean protein-berberine as the research object, flux, transmittance of berberine and traping rate of protein as indexes, different solution environment on membrane process were investigated.</p><p><b>RESULT</b>When the concentration of soybean protein was under 1 g x L(-1), the membrane flux was minimum with the traping of berberine decreased slightly as the concentration increased. When pH was 4, the flux was maximum with the traping rate of protein was 99%, and the transmittance of berberine reached above 60%.</p><p><b>CONCLUSION</b>The efficiency of membrane separation can be improved by optimizing the solution environment of water-extraction of chinese medicines. The efficiency of membrane separation is the best when adjust the pH to the isoelectric point of proteins for the proteins as the main pollutant in aqueous solution.</p>

Effects of PPAR-α silence on ET-1 induced myocardial hypertrophy and PI3K/Akt/GSK3β-NFATc4 signal pathway / 中国病理生理杂志

AIM: To investigate the role and signal mechanism of PPAR-α in the pathogenesis of cardiac hypertrophy. METHODS: Small interfering RNA (siRNA) was applied to efficiently silence the gene expression of PPAR-α in cardiac myocytes. [~3H] leucine incorporation assay was performed to measure protein synthesis. Reverse transcription-polymerase chain reaction (RT-PCR) was used to analyze the mRNA level of atrial natriuretic factor (ANF) and PPAR-α. Western blotting analysis was performed to investigate the levels of phosphorylation of protein kinase B (PKB/Akt) and glycogen synthase kinase 3β (GSK3β). Immunofluorescence analysis was used to examine the cellular localization of NFATc4. RESULTS: (1)RSS304168 was the most efficient stealth RNAi duplex to specifically inhibit PPAR-α expression. (2)RSS304168 significantly potentiated the ET-1-induced cardiomyocyte hypertrophy and enhanced ET-1-induced protein synthesis and ANF mRNA expression in cardiomyocytes. Moreover, RSS304168 completely reversed the inhibitory effects of fenofibrate on ET-1-induced protein synthesis and ANF mRNA expression. (3)RSS304168 enhanced ET-1-induced phosphorylation of Akt at Ser473 and GSK3β at Ser9. The effects of ET-1 or ET-1 combined with RSS304168 on phosphorylation of Akt/GSK3β were completely blocked by LY294002, a PI3K specific inhibitor. Fenofibrate markedly inhibited ET-1-induced phosphorylation of Akt/GSK3β while RSS304168 abolished these effects of fenofibrate. (4)Fenofibrate prevented the nuclear translocation of NFATc4 induced by ET-1 while RSS304168 abolished this effect of fenofibrate. CONCLUSION: Activation of PPAR-α inhibits ET-1-induced cardiomyocyte hypertrophy through blocking Akt/GSK3β-NFATc4 signaling pathways.

Antennal sensilla of grasshoppers (Orthoptera: Acrididae) in relation to food preferences and habits.

The external structure, i.e. number and distribution of sensillae on male and female antennae of 12 species of grasshoppers belonging to Pamphaginae, Catantopinae, Oedipodinae and Gomphocerinae in the grasslands of Inner Mongolia was investigated using scanning electron microscopy. Five major types of antennal sensillae were detected--trichoid, long basiconic, short basiconic, slender and short basiconic, and coeloconic sensillae. Total number of antennal sensillae varied among different sexes, subfamilies, feeding groups, life forms and eco-forms. Males showed significantly more sensillae than females, due to presence of more short basiconic and coeloconic sensillae. Species under Catantopinae showed more long basiconic sensillae than the others. The Oedipodinae had the highest number of slender and short basiconic sensillae and coeloconic sensillae, followed by Catantopinae and Gomphocerinae; while Pamphaginae had the fewest. The total number of sensillae showed the same trend for these two types amongst the subfamilies as well, species which prefer habits on the ground possessed fewer antennal sensillae than species which prefer to stay on plants. The maximal number of antennal sensillae were observed in hygrophytous species, Chorthippus albomarginatus, in the 12 grasshopper species investigated, although the data is not statistically significant. The general trend which emerged was that species feeding on grass possessed more antennal sensillae, particularly coeloconic sensillae, compared to other feeding group species.