ObjectiveTo investigate the effects and potential mechanisms of Babaodan （BBD） against acetaminophen （APAP）-induced acute liver injury （ALI） based on transcriptomics. MethodsA total of 36 male C57BL/6 mice were randomly divided into 6 groups （n=6 per group）： normal group， model group， N-acetylcysteine group （NAC， 120 mg·kg^-1）， and BBD low-， medium-， and high-dose groups （BBD-L， BBD-M， BBD-H groups， 75， 150， 300 mg·kg^-1， respectively）. Except for the normal group， all other groups were subjected to APAP-induced ALI. The serum levels of aspartate aminotransferase （AST）， alanine aminotransferase （ALT）， total cholesterol （TC）， triglyceride （TG）， and low-density lipoprotein cholesterol （LDL-C） were measured in each group. Hepatic levels or activities of malondialdehyde （MDA） and glutathione peroxidase （GSH-Px） were detected using commercial kits. Hematoxylin-eosin （HE） staining was performed to evaluate the degree of liver histopathological damage. Transcriptomic analysis was employed to screen differentially expressed genes （DEGs）， and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis was performed to identify differential pathways involved in BBD intervention against ALI. Real-time quantitative polymerase chain reaction （Real-time PCR） and Western blot were applied to validate the expression of differential genes and related pathway proteins. Additionally， glucose （GLU） consumption， as well as lactate （LD） and adenosine triphosphate （ATP） content were assessed across all mouse groups. ResultsPharmacodynamic evaluation showed that， compared with the normal group， the model group exhibited significantly elevated serum levels of ALT， AST， TC， TG， and LDL-C （P<0.01）， significantly increased MDA level （P<0.01）， and significantly decreased GSH-Px level （P<0.05）. Compared with the model group， BBD intervention at different doses significantly reduced the serum levels of ALT， AST， TC， TG， and LDL-C （P<0.05， P<0.01）， increased GSH-Px level （P<0.05， P<0.01）， significantly decreased MDA level （P<0.01）， and ameliorated hepatic histopathological injury. Liver transcriptomic analysis revealed that， following high-dose BBD intervention， the core genes were mainly enriched in pathways related to inflammatory responses and energy metabolic reprogramming， including the interleukin-17 （IL-17） signaling pathway， the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway， fructose and mannose metabolism， and glycolysis. Real-time PCR validation demonstrated that， compared with the normal group， the mRNA expression levels of glycolysis-related genes ［hexokinase 1 （HK1）， hexokinase 2 （HK2）， 6-phosphofructo-2-kinase/fructose-2，6-bisphosphatase 3 （PFKFB3）， 6-phosphofructo-2-kinase/fructose-2，6-bisphosphatase 4 （PFKFB4）， pyruvate kinase M （PKM）， and lactate dehydrogenase A （LDHA）］， as well as inflammatory cytokines ［tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， and interleukin-1β （IL-1β）］ were significantly upregulated in the model group （P<0.05， P<0.01）. Compared with the model group， the BBD-H group showed significantly decreased mRNA expression of HK1， HK2， PFKFB3， PFKFB4， PKM， LDHA， TNF-α， IL-6， and IL-1β （P<0.05， P<0.01）. Western blot results indicated that， compared with the normal group， the model group had significantly increased expression of glycolysis-related proteins ［glucose transporter 1 （GLUT1）， HK1， PFKFB3， and PKM］， inflammatory proteins ［interleukin-18 （IL-18）， TNF-α， and IL-1β］， and phosphorylated （p）-PI3K and p-Akt proteins （P<0.05， P<0.01）. Compared with the model group， the BBD-H group exhibited significantly decreased expression of GLUT1， HK1， PFKFB3， PKM， IL-18， TNF-α， IL-1β， p-PI3K， and p-Akt （P<0.05， P<0.01）. Metabolic indicator measurements showed that， compared with the model group， the BBD-H group showed significantly reduced GLU consumption， LD and ATP content （P<0.05， P<0.01）. ConclusionBBD may alleviate APAP-induced ALI through dual regulation of metabolic reprogramming and inflammatory responses， potentially via inhibition of the PI3K/Akt signaling pathway.

ObjectiveTo investigate the effects and potential mechanisms of Babaodan （BBD） against acetaminophen （APAP）-induced acute liver injury （ALI） based on transcriptomics. MethodsA total of 36 male C57BL/6 mice were randomly divided into 6 groups （n=6 per group）： normal group， model group， N-acetylcysteine group （NAC， 120 mg·kg^-1）， and BBD low-， medium-， and high-dose groups （BBD-L， BBD-M， BBD-H groups， 75， 150， 300 mg·kg^-1， respectively）. Except for the normal group， all other groups were subjected to APAP-induced ALI. The serum levels of aspartate aminotransferase （AST）， alanine aminotransferase （ALT）， total cholesterol （TC）， triglyceride （TG）， and low-density lipoprotein cholesterol （LDL-C） were measured in each group. Hepatic levels or activities of malondialdehyde （MDA） and glutathione peroxidase （GSH-Px） were detected using commercial kits. Hematoxylin-eosin （HE） staining was performed to evaluate the degree of liver histopathological damage. Transcriptomic analysis was employed to screen differentially expressed genes （DEGs）， and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis was performed to identify differential pathways involved in BBD intervention against ALI. Real-time quantitative polymerase chain reaction （Real-time PCR） and Western blot were applied to validate the expression of differential genes and related pathway proteins. Additionally， glucose （GLU） consumption， as well as lactate （LD） and adenosine triphosphate （ATP） content were assessed across all mouse groups. ResultsPharmacodynamic evaluation showed that， compared with the normal group， the model group exhibited significantly elevated serum levels of ALT， AST， TC， TG， and LDL-C （P<0.01）， significantly increased MDA level （P<0.01）， and significantly decreased GSH-Px level （P<0.05）. Compared with the model group， BBD intervention at different doses significantly reduced the serum levels of ALT， AST， TC， TG， and LDL-C （P<0.05， P<0.01）， increased GSH-Px level （P<0.05， P<0.01）， significantly decreased MDA level （P<0.01）， and ameliorated hepatic histopathological injury. Liver transcriptomic analysis revealed that， following high-dose BBD intervention， the core genes were mainly enriched in pathways related to inflammatory responses and energy metabolic reprogramming， including the interleukin-17 （IL-17） signaling pathway， the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway， fructose and mannose metabolism， and glycolysis. Real-time PCR validation demonstrated that， compared with the normal group， the mRNA expression levels of glycolysis-related genes ［hexokinase 1 （HK1）， hexokinase 2 （HK2）， 6-phosphofructo-2-kinase/fructose-2，6-bisphosphatase 3 （PFKFB3）， 6-phosphofructo-2-kinase/fructose-2，6-bisphosphatase 4 （PFKFB4）， pyruvate kinase M （PKM）， and lactate dehydrogenase A （LDHA）］， as well as inflammatory cytokines ［tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， and interleukin-1β （IL-1β）］ were significantly upregulated in the model group （P<0.05， P<0.01）. Compared with the model group， the BBD-H group showed significantly decreased mRNA expression of HK1， HK2， PFKFB3， PFKFB4， PKM， LDHA， TNF-α， IL-6， and IL-1β （P<0.05， P<0.01）. Western blot results indicated that， compared with the normal group， the model group had significantly increased expression of glycolysis-related proteins ［glucose transporter 1 （GLUT1）， HK1， PFKFB3， and PKM］， inflammatory proteins ［interleukin-18 （IL-18）， TNF-α， and IL-1β］， and phosphorylated （p）-PI3K and p-Akt proteins （P<0.05， P<0.01）. Compared with the model group， the BBD-H group exhibited significantly decreased expression of GLUT1， HK1， PFKFB3， PKM， IL-18， TNF-α， IL-1β， p-PI3K， and p-Akt （P<0.05， P<0.01）. Metabolic indicator measurements showed that， compared with the model group， the BBD-H group showed significantly reduced GLU consumption， LD and ATP content （P<0.05， P<0.01）. ConclusionBBD may alleviate APAP-induced ALI through dual regulation of metabolic reprogramming and inflammatory responses， potentially via inhibition of the PI3K/Akt signaling pathway.

Objective:To investigate the effect of overexpression of T cell restricted intracellular antigen 1(TIA1)gene in M2-type tumor-associated macrophages(M2-TAMs)on invasion and migration of gastric cancer cells and its mechanism.Methods:Primary TAMs were extracted from gastric cancer tissues and induced to differentiate into M2-TAMs using IL-4 and IL-13.The TIA1 overex-pression plasmid(oe-TIA1)and its empty vector were transfected into M2-TAMs.The expression levels of TIA1 mRNA and protein were detected by qRT-PCR and Western blot.The expression levels of CD206 and CD163 were detected by flow cytometry.The levels of IL-10,TGF-β,VEGF-A and Arg-1 in cell culture supernatant were detected by ELISA.The transfected M2-TAMs were co-cultured with gastric cancer BGC-823 cells by Transwell co-cultivation system,and PI3K agonist 740Y-P was used to intervene in parallel.The cell migration ability was detected by scratch assay.Transwell assay was used to detect cell invasion ability.Protein expression levels of PI3K,p-PI3K,AKT,p-AKT,MMP-2 and MMP-9 in the cells were detected by Western blot.Results:①Compared with primary TAMs,the levels of CD206 and CD163 expression in M2-TAMs cells and IL-10,TGF-β,VEGF-A and Arg-1 in cell culture superna-tant were significantly increased(P<0.05),while the expression levels of TIA1 mRNA and protein were significantly decreased(P<0.05).Overexpression of TIA1 gene significantly decreased the expression levels of CD206 and CD163 in M2-TAMs and IL-10,TGF-β,VEGF-A and Arg-1 in cell culture supernatant(P<0.05).②Overexpression of TIA1 gene in M2-TAMs could significantly reduce the migration and invasion ability and the expression levels of p-PI3K/PI3K,p-AKT/AKT,MMP-2 and MMP-9 proteins in BGC-823 cells(P<0.05).③740Y-P could significantly reverse the inhibitory effects of overexpression of TIA1 gene in M2-TAMs on migration,inva-sion and PI3K/AKT signaling pathway of BGC-823 cells.Conclusion:Overexpression of TIA1 gene in M2-TAMs can affect the inva-sion and migration of gastric cancer cells by blocking the PI3K/AKT signaling pathway.

Objective:To discuss the inhibitory effect of microRNA-487a(miR-487a)on the M2 polarization of tumor-associated macrophages(TAMs)in gastric cancer,and to clarify its effect on the proliferation,invasion,and migration of the gastric cancer AGS cells.Methods:The TAMs from gastric cancer tissue and adjacent normal tissue macrophages(NTMs)from adjacent tissue of the primary gastric cancer patients were isolated and cultured.The human monocyte THP-1 cells were induced in vitro to differentiate into TAMs,and the differentiated M0,M1,and M2 macrophages were cultured for 24 h by conditioned medium(CM)to obtain the TAMs,M1-TAMs,and M2-TAMs respectively.The TAMs were transfected and then divided into blank group,inhibitor-NC group,miR-487a inhibitor group,miR-487a inhibitor+si-NC group,and miR-487a inhibitor+si-TIA1 group.The transfection efficiencies of the cells in various groups were detected by real-time fluorescence quantitative PCR(RT-qPCR)and Western blotting methods.The M2-TAMs were co-cultured with the AGS cells,and divided into AGS group,AGS+inhibitor-NC group,AGS+miR-487a inhibitor group,AGS+miR-487a inhibitor+si-NC group,and AGS+miR-487a inhibitor+si-TIA1 group.RT-qPCR method was used to detect the expression levels of miR-487a and lymphocyte intracytoplasmic antigen-1(TIA1)mRNA in TAMs from gastric cancer tissue and NTMs from adjacent normal tissue in various groups;Western blotting method was used to detect the expression level of TIA1 protein in TAMs from gastric cancer tissue and NTMs from adjacent normal tissue and TAMs in various groups;flow cytometry was used to detect the levels of CD206 and CD163 in TAMs in various groups;enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of interleukin-10(IL-10),transforming growth factor-beta(TGF-β),vascular endothelial growth factor A(VEGF-A),and arginase-1(Arg-1)in culture supernatant of the TAMs cells;CCK-8 assay was used to detect the proliferative activity of the AGS cells in various groups;wound healing assay was used to detect the migration rates of the AGS cells in various groups;Transwell assay was used to detect the number of invasion AGS cells in various groups.Results:The RT-qPCR results shoued that compared with NTMs from adjacent tissue,the expression level of miR-487a in the TAMs from gastric cancer tissue was significantly increased(P<0.01)and the expression level of TIA1 mRNA was significantly decreased(P<0.01).Compared with TAMs,the expression level of miR-487a in M1-TAMs was significantly decreased(P<0.01),and the expression level of TIA1 mRNA was increased(P<0.01);the expression level of miR-487a in M2-TAMs was significantly increased(P<0.01),and the expression level of TIA1 mRNA was decreased(P<0.01).After transfection,compared with blank group and inhibitor-NC group,the expression level of miR-487a in the cells in miR-487a inhibitor group was significantly decreased(P<0.01),indicating successful transfection.The Western blotting results showed that compared with NTMs from adjacent normal tissue,the expression level of TIA1 protein in TAMs from gastric cancer tissue was decreased(P<0.01);compared with TAMs,the expression level of T1A1 protein in M1-TAMs was significantly increased(P<0.01),and the expression of TIA1 protein in M2-TAMs was significantly decreased(P<0.01);after co-transfection,compared with inhibitor-NC group,the expression level of TIA1 protein in the cells in miR-487a inhibitor group was significantly increased(P<0.01);compared with miR-487a inhibitor+si-NC group,the expression level of TIA1 protein in the cells in miR-487a inhibitor+si-TIA1 group was significantly decreased(P<0.01).The flow cytometry results showed that compared with blank group and inhibitor-NC group,the levels of CD206 and CD163 in the cells in miR-487a inhibitor group were significantly decreased(P<0.01);after co-transfection,compared with inhibitor-NC group,the levels of CD206 and CD163 in the cells in miR-487a inhibitor group were significantly decreased(P<0.01);compared with miR-487a inhibitor+si-NC group,the levels of CD206 and CD163 in the cells in miR-487a inhibitor+si-TIA1 group were significantly increased(P<0.01).The ELISA results showed that compared with blank group and inhibitor-NC group,the levels of IL-10,TGF-β,VEGF-A,and Arg-1 in culture supernatant of the TAMs in miR-487a inhibitor group were significantly decreased(P<0.01);after co-transfection,compared with inhibitor-NC group,the levels of IL-10,TGF-β,VEGF-A,and Arg-1 in culture supernatant of the TAMs in miR-487a inhibitor group were significantly decreased(P<0.01);compared with miR-487a inhibitor+si-NC group,the levels of IL-10,TGF-β,VEGF-A,and Arg-1 in culture supernatant of the TAMs in miR-487a inhibitor+si-TIA1 group were significantly increased(P<0.01).The CCK-8 assay results showed that compared with AGS group,the proliferation activity of the cells in AGS+inhibitor-NC group was significantly increased(P<0.01);compared with AGS+inhibitor-NC group,the proliferation activity of the cells in AGS+miR-487a inhibitor group was significantly decreased(P<0.01);compared with AGS+miR-487a inhibitor+si-NC group,the proliferation activity of the cells in AGS+miR-487a inhibitor+si-TIA1 group was significantly increased(P<0.01).The wound healing assay results showed that compared with AGS group,the migration rate of the cells in AGS+inhibitor-NC group was significantly(P<0.05);compared with AGS+inhibitor-NC group,the migration rate of the cells in AGS+miR-487a inhibitor group was significantly decreased(P<0.01);compared with AGS+miR-487a inhibitor+si-NC group,the migration rate of the cells in AGS+miR-487a inhibitor+si-TIA1 group was significantly increased(P<0.05).The Transwell assay results showed that compared with AGS group,the number of invasion AGS cells in AGS+inhibitor-NC group was significantly increased(P<0.01);compared with AGS+inhibitor-NC group,the number of invasion AGS cells in AGS+miR-487a inhibitor group was significantly decreased(P<0.01);compared with AGS+miR-487a inhibitor+si-NC group,the number of invasion AGS cells in AGS+miR-487a inhibitor+si-TIA1 group was significantly increased(P<0.01).Conclusion:Silencing the miR-487a expression can inhibit the M2 polarization of the gastric cancer-associated macrophages by targeted upregulation of TIA1,and suppress the proliferation,migration,and invasion of the gastric cancer cells.

OBJECTIVE To investigate the improvement effects and possible mechanism of Phyllanthus emblica water extracts on lipid metabolism disorder and insulin resistance （IR） in diabetic model rats. METHODS IR model of male SD rats was established and randomly divided into model group， positive control group， P. emblica water extracts high-dose， medium-dose and low-dose groups， and another blank group was set up， with 10 rats in each group. P. emblica water extracts high-dose， medium- dose and low-dose groups were given P. emblica water extract diluent at the doses of 800， 400 and 200 mg/kg respectively； positive control group was given pioglitazone solution 2.7 mg/kg； blank group and model group were given the same volume of normal saline， by intragastrical administration， once a day， for consecutive 4 weeks. The general state of rats was observed， and the body mass and the levels of serum lipid metabolism indexes [triacylglycerol （TG）， total cholesterol （TC）， low-density lipoprotein cholesterol （LDL-C）， high-density lipoprotein cholesterol （HDL-C）] were measured； fasting insulin of serum （FINS）， insulin resistance index （IRI） and insulin sensitivity index （ISI） were measured and calculated by enzyme-linked immunosorbent assay （ELISA）. The pathological morphology of liver tissue was observed by hematoxylin-eosin staining； the expression of glycogen in the liver was observed by periodic acid-Schiff staining， and the staining area was calculated； the protein expressions of peroxisome proliferator-activated receptor γ （PPARγ）， nuclear factor κB（NF-κB） and AMP-activated protein kinase （AMPK） were detected by Western blot. RESULTS Compared with blank group， the rats in the model group had obvious symptoms of polydipsia， polyphagia and polyuria， and the serum levels of TC， TG， LDL-C， FINS and IRI were significantly increased （P＜0.05）， while ISI was significantly reduced （P＜ 0.05）； the structure of hepatic lobule was obviously disordered， the liver cells were deformed and enlarged， and there were many lipid deposits and fat vacuoles； PAS staining area of glycogen was significantly reduced（P＜0.05）； the protein expression levels of PPARγ and p-AMPK were significantly decreased （P＜0.05）， while the protein expression level of NF-κB was significantly increased （P＜0.05）. Compared with model group， the mental state of rats， liver histopathological morphology and glycogen expression were improved significantly in P. emblica water extracts high-dose and medium-dose groups； the levels of the above indicators were significantly reversed（P＜0.05）. CONCLUSIONS P. emblica water extracts may improve glucose and lipid metabolism disorders， liver function and IR in IR model rats， and regulate IR by activating the PPARγ/AMPK/NF-κB signaling pathway.

Until the recent emergence/re-emergence of human-pathogenic viruses in ticks, tick-borne viruses have been neglected as causative agents of human disease (particularly in China). To gain insight into the diversity of tick-borne viruses in Xinjiang Uygur Autonomous Region (northwestern China), we conducted illumina deep sequencing-based screening for virus-derived small RNAs in field-collected Ixodes persulcatus ticks. We found 32, 631 unique virus-matched reads. In particular, 77 reads mapped to the tick-borne group within the genus of Flavivirus, and covered 3.8%-2.4% viral genomes. In addition, 32 unique reads were specific to the Siberian subtype of tick-borne encephalitis viruses (TBEV-Sib) which have never been reported in Chinese TBE loci. We confirmed the potential existence of TBEV-Sib by amplification (using reverse transcription-polymerase chain reaction) of genomic fragments from the envelope gene or 3' genomic terminus from the pools of examined ticks. Both sequences demonstrated high homology to TBEV-Sib strains attached geographically to southern Siberia with nucleotide identity of 97.2%-95.5% and aminoacid identity of 99.4%-98.3%, respectively. In conclusion, we report, for the first time, detection of TBEV-Sib in the natural TBE loci of China. These novel data may provide genetic information for further isolation and epidemiologic investigation of TBEV-Sib.
Animals ; Arachnid Vectors ; virology ; China ; Encephalitis Viruses, Tick-Borne ; classification ; genetics ; isolation & purification ; Encephalitis, Tick-Borne ; transmission ; virology ; Genome, Viral ; Humans ; Ixodes ; virology ; Molecular Sequence Data ; Phylogeny