Parthenolide inhibits the proliferation and induces the apoptosis of human uveal melanoma cells

AIM: To explore the effect of parthenolide(PTL) on human uveal melanoma(UM) cells(C918 and SP6.5 cells) and its molecular mechanism. METHODS: Carboxyfluorescein succinimidyl amino ester(CFSE) assays and cell counting kit-8(CCK-8) were performed to detect the cell viability. Flow cytometry was used to analyze cell cycle and apoptosis. Quantitative realtime polymerase chain reaction(qRT-PCR) and Western blot assays were performed to measure proliferation-related and apoptosis-related factors.RESULTS: Firstly, PTL decreased the viability of C918 and SP6.5 cells in a dose-dependent manner, and the effect of PTL on C918 cells was stronger than on SP6.5;however, it did not affect normal cells. Secondly, PTL increased the proportion of cell number at cell cycle G1 phase in C918 cells, and decreased the proportion of cell number at S phase, but the proportion did not change at G2 phase. In addition, PTL induced the apoptosis of C918 cells, and decreased the expressions of Cyclin D1, B-cell lymphoma-2(Bcl-2) and B-cell lymphoma-extra large(Bcl-XL). Also, PTL increased Cyclin inhibition protein 1(P21), Bcl-2-associated X protein(Bax), Cysteinyl aspartate specific proteinas-3(Caspase-3) and Caspase-9 expression. However, the expression of Caspase-8 was not changed. CONCLUSION: PTL inhibites proliferation and induces apoptosis in UM cells by arresting G1 phase and regulating mitochondrial pathway, however, it does not affect normal cells.

Cloning and Bioinformatics Analysis of TpiA Gene of Vibrio alginolyticus HY9901

[Objectives]To clone and analyze the TpiA gene of Vibrio alginolyticus HY9901.[Methods]According to the TpiA gene sequence of V.alginolyticus,a pair of specific primers was designed,and its full length was amplified by PCR.[Results]The full length of TpiA gene is 771 bp,encoding 256 amino acid residues in total,and the NCBI accession number is OM906798.According to the deduced amino acid sequence,its molecular weight was predicted to be about 26.97548 kDa,and its isoelectric point was 4.78.The amino acid sequence of the N-terminal signal peptide structure was predicted,and it was found that there was no obvious signal peptide cleavage site,no signal peptide,and no transmembrane region;the amino acid sequence contained 3 N-glycosylation sites,4 protein kinase C phosphorylation sites,2 casein kinase II phosphorylation sites,6 N-myristoylation sites,7 microbody C-terminal target signal site,and 1 triose phosphate isomerase active site.The prediction results of protein subcellular localization showed that TpiA may be located in mitochondria or cytoplasm,with probability of 39.1%and 34.8%,respectively.The amino acid sequence of the TpiA gene of V.alginolyticus shared 98.83%-99.61%homology with other Vibrio species,and it was clustered into the same subfamily with Vibrio parahaemolyticus and had a close relationship.In the secondary structure prediction,the proportions ofα-helix,random coil and extended chain were 44.53%,41.41%and 14.06%,respectively,and the similarity of its tertiary structure model to template 1aw1.1.A was 85.16%.[Conclusions]This study is intended to provide a basis for further research on the role of TpiA gene in the type III secretion system and related research on antibiotic resistance.

The Fibrillin-1/VEGFR2/STAT2 signaling axis promotes chemoresistance via modulating glycolysis and angiogenesis in ovarian cancer organoids and cells

Background:Chemotherapy resistance is a primary reason of ovarian cancer therapy failure;hence it is important to investigate the underlying mechanisms of chemotherapy resistance and develop novel potential therapeutic targets.Methods:RNA sequencing of cisplatin-resistant and sensitive(chemoresis-tant and chemosensitive,respectively)ovarian cancer organoids was performed,followed by detection of the expression level of fibrillin-1(FBN1)in organoids and clinical specimens of ovarian cancer.Subsequently,glucose metabolism,angiogenesis,and chemosensitivity were analyzed in structural glycoprotein FBNl-knockout cisplatin-resistant ovarian cancer organoids and cell lines.To gain insights into the specific functions and mechanisms of action of FBN1 in ovarian cancer,immunoprecipitation,silver nitrate staining,mass spectrometry,immunofluorescence,Western blotting,and Forister resonance energy transfer-fluorescence lifetime imaging analyses were performed,followed by in vivo assays using vertebrate model systems of nude mice and zebrafish.Results:FBN1 expression was significantly enhanced in cisplatin-resistant ovar-ian cancer organoids and tissues,indicating that FBNI might be a key factor in chemoresistance of ovarian cancer.We also discovered that FBN1 sustained the energy stress and induced angiogenesis in vitro and in vivo,which promoted the cisplatin-resistance of ovarian cancer.Knockout of FBN1 combined with treat-ment of the antiangiogenic drug apatinib improved the cisplatin-sensitivity of ovarian cancer cells.Mechanistically,FBN1 mediated the phosphorylation of vascular endothelial growth factor receptor 2(VEGFR2)at the Tyrl054 residue,which activated its downstream focal adhesion kinase(FAK)/protein kinase B(PKB or AKT)pathway,induced the phosphorylation of signal transducer and activator of transcription 2(STAT2)at the tyrosine residue 690(Tyr690),pro-moted the nuclear translocation of STAT2,and ultimately altered the expression of genes associated with STAT2-mediated angiogenesis and glycolysis.Conclusions:The FBN1/VEGFR2/STAT2 signaling axis may induce chemore-sistance of ovarian cancer cells by participating in the process of glycolysis and angiogenesis.The present study suggested a novel FBNl-targeted therapy and/or combination of FBN1 inhibition and antiangiogenic drug for treating ovarian cancer.

Herbal formula BaWeiBaiDuSan alleviates polymicrobial sepsis-induced liver injury via increasing the gut microbiota Lactobacillus johnsonii and regulating macrophage anti-inflammatory activity in mice

Sepsis-induced liver injury(SILI)is an important cause of septicemia deaths.BaWeiBaiDuSan(BWBDS)was extracted from a formula of Panax ginseng C.A.Meyer,Lilium brownie F.E.Brown ex Miellez var.viridulum Baker,Polygonatum sibiricum Delar.ex Redoute,Lonicera japonica Thunb.,Hippophae rhamnoides Linn.,Amygdalus Communis Vas,Platycodon grandiflorus(Jacq.)A.DC.,and Cortex Phelloderdri.Herein,we investigated whether the BWBDS treatment could reverse SILI by the mechanism of modulating gut microbiota.BWBDS protected mice against SILI,which was associated with promoting macrophage anti-inflammatory activity and enhancing intestinal integrity.BWBDS selectively promoted the growth of Lactobacillus johnsonii(L.johnsonii)in cecal ligation and puncture treated mice.Fecal microbiota transplantation treatment indicated that gut bacteria correlated with sepsis and was required for BWBDS anti-sepsis effects.Notably,L.johnsonii significantly reduced SILI by promoting macrophage anti-inflammatory activity,increasing interleukin-10+M2 macrophage production and enhancing intestinal integrity.Furthermore,heat inactivation L.johnsonii(HI-L.johnsonii)treatment promoted macrophage anti-inflammatory activity and alleviated SILI.Our findings revealed BWBDS and gut microbiota L.johnsonii as novel prebiotic and probiotic that may be used to treat SILI.The potential underlying mechanism was at least in part,via L.johnsonii-dependent immune regulation and interleukin-10+M2 macrophage production.

Mechanism of Wuweijiangyasan in the Treatment of Spontaneous Hypertension based on Network Pharmacology

Background:Hypertension affects over 1 billion people globally and is the top risk factor of cardiovascular morbidity and mortality.Wuweijiangyasan(WWJYS),as an empirical prescription,has stable depressurization effects.This study investigated the chemical composition and pharmacodynamic effects of WWJYS in regulating the blood pressure(BP),emotion,and blood lipid of spontaneous hypertensive rats,and further explored the depressurization mechanism of WWJYS.Materials and Methods:This study used network pharmacology to identify the origins and predict targets of WWJYS,and artificial intelligence-based molecular docking is used to further predict targets and mechanisms.The chemical constituents of WWJYS were analyzed and identified by ultra high-performance liquid chromatography–mass spectrometry(MS)/MS.Results:In the WWJYS group,the systolic BP level significantly was decreased,and the HR was stable.The irritability became stable after the 5-week treatment compared with the model group(P<0.05).Rats’rotation tolerance time increased after 2-weeks stabilization.Compared with the model group,angiotensin-converting enzyme 2 protein and mRNA of the WWJYS group increased significantly(P<0.05).Network pharmacology collected 64 compounds and identified 22 potential targets of WWJYS for antihypertensive activity.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis showed that WWJYS might regulate smooth muscle cells,affect inflammatory response and improve endothelial function through multiple pathways.The molecular docking study further supported that the target proteins have good combinations with the main active components of WWJYS.Conclusions:The data indicated that WWJYS had significant depressurization,analgesic,and sedative,as well as lipid-lowering effects,and the depressurization mechanism of WWJYS may function in multiple signal pathways,especially in improving blood vessel function and intervening inflammation.

Author correction to‘Herbal formula BaWeiBaiDuSan alleviates polymicrobial sepsisinduced liver injury via increasing the gut microbiota Lactobacillus johnsonii and regulating macrophage anti-inflammatory activity in mice'[Acta Pharmaceutica Sinica B 13(2023)1164-1179]

The authors regret that there were some picture errors in Fig.7C and Supporting Information Fig.S10B owing to the negligence of the picture typesetting and careless mistakes.In Fig.7C,the H&E picture of PBS+Lipo+CLP group was the inverted picture of CLP group in Fig.5G.In Fig.7C,the H&E picture of Clo-Lipo+CLP group was zoom-in picture of L johnsoni+CLP group in Fig.8F.In Fig.SI0B,the H&E picture of ileum in CLP group was zoom-in picture of Anti-IL-10R+CLP group in Fig.S11C.The authors revise the H&E picture of liver in PBS+Lipo+CLP group and the H&E picture of liver in Clo-Lipo+CLP group in Fig.7C.Also,the H&E picture of ileum in CLP group of Fig.S10B have been revised.The correct figures are presented as below.