The culture of suspended Vero cells is facing difficulties such as low cell viability and long doubling time. To investigate the main reasons for the slow growth and low viability of suspended Vero cells, this study conducted transcriptomic analysis of suspended Vero cells (Vero-XF) and adherent Vero cells (Vero-AD) to screen the differentially expressed genes (DEGs) affecting the growth of suspended cells. In addition, epidermal growth factor (EGF) was supplemented to the culture system to improve the growth of Vero-XF. The results showed that compared with the Vero-AD group, the Vero-XF group had 7 376 significant DEGs. Kyoto encyclopedia of genes and genomes enrichment analysis revealed that the DEGs were mainly enriched in the autophagy and mitophagy pathways. Eleven DEGs were selected and verified by quantitative real-time PCR, which showed up-regulated expression of ATG9B, WIPI2, LAMP2, OPTN, Rab7a, and DEPTOR and down-regulated expression of ATG4D, being consistent with the results of transcriptomic analysis. In addition, the Vero-XF group showed significantly up-regulated expression of ATG101, ATG2A, and STX17 and insignificant change in the expression of NBR1, compared with the Vero-AD group. The protein levels of LC3 and P62 in Vero-XF and Vero-AD were determined by Western blotting, which showed up-regulated expression of LC3Ⅱ/Ⅰ and down-regulated expression of P62 in Vero-XF, indicating a higher level of autophagy. Finally, the exogenous supplementation of EGF at 10, 20, and 30 μg/L in the culture system reduced the autophagy level of Vero-XF by 22.35%, 48.15%, and 71.29%, increased the specific growth rate by 15.48%, 33.33%, and 57.14%, and decreased the apoptosis rate by 2.84%, 15.46%, and 16.23%, respectively. The results of this study preliminarily reveal that the activation of autophagy is one of the reasons for the slow growth of Vero-XF, which provides reference for the subsequent culture of suspended Vero cells.
Animals ; Vero Cells ; Autophagy/genetics* ; Chlorocebus aethiops ; Epidermal Growth Factor/pharmacology* ; Gene Expression Profiling ; Transcriptome ; Cell Survival

Objective:To explore the effects of anshen jieyu decoction on hippocampal morphology and regulation of PI3K/AKT signalling pathway with hypothalamic-pituitary-adrenal axis expression in CUMS depressed rats.Methods:Rats were divided into a control group(Control),CUMS model group(CUMS),and ASJYD treatment group(CUMS+ASJYD).Rats in the CUMS and CUMS+ASJYD groups were exposed to 10 different chronic stressors to induce depressive-like behaviors.After modeling,the CUMS+ASJYD group received ASJYD via gavage.Depressive-like behaviors were assessed using the sucrose preference test and open field test.Serum levels of adrenocorticotropic hor-mone(ACTH)and corticosterone(CORT)were measured by ELISA.Hippocampal neuronal morphological changes were observed via Nissl staining,mitochondrial ultrastructure was examined using transmission electron microscopy,and hippocampal PI3K and AKT protein phosphorylation levels were detected by Western blot.Results:Compared with the Control group,the CUMS group showed significantly reduced sucrose consumption,preference rate,total travel distance in the open field,and central zone activity time(P＜0.01).Serum ACTH and CORT levels were elevated(P＜0.01),with disorganized hippocampal cell arrangement,reduced cell count,mitochondrial swelling,cristae blurring/rupture observed under electron microscopy,and decreased phosphorylation levels of PI3 K and AKT proteins(P＜0.01).After ASJYD treatment,the CUMS+ASJYD group exhibited significant improvements in sucrose consumption,preference rate,locomotor activity,and central zone exploration(P＜0.01),accompanied by reduced ACTH/CORT expression,alleviated hippocampal pathology,restored mitochondrial integrity with clear cristae,and increased PI3K/AKT phosphorylation(P＜0.01).Conclusion:Anshen Jieyu decoction significantly improved the depression-like be-haviour of CUMS rats,which may be achieved by down-regulating the hyperactive hypothalamic-pituitary-adrenal axis,regulating the expression of PI3K/AKT signaling pathway,and ameliorating the damage to hippocampal neurons and mi-tochondria.

Objective:To explore the effects of anshen jieyu decoction on hippocampal morphology and regulation of PI3K/AKT signalling pathway with hypothalamic-pituitary-adrenal axis expression in CUMS depressed rats.Methods:Rats were divided into a control group(Control),CUMS model group(CUMS),and ASJYD treatment group(CUMS+ASJYD).Rats in the CUMS and CUMS+ASJYD groups were exposed to 10 different chronic stressors to induce depressive-like behaviors.After modeling,the CUMS+ASJYD group received ASJYD via gavage.Depressive-like behaviors were assessed using the sucrose preference test and open field test.Serum levels of adrenocorticotropic hor-mone(ACTH)and corticosterone(CORT)were measured by ELISA.Hippocampal neuronal morphological changes were observed via Nissl staining,mitochondrial ultrastructure was examined using transmission electron microscopy,and hippocampal PI3K and AKT protein phosphorylation levels were detected by Western blot.Results:Compared with the Control group,the CUMS group showed significantly reduced sucrose consumption,preference rate,total travel distance in the open field,and central zone activity time(P＜0.01).Serum ACTH and CORT levels were elevated(P＜0.01),with disorganized hippocampal cell arrangement,reduced cell count,mitochondrial swelling,cristae blurring/rupture observed under electron microscopy,and decreased phosphorylation levels of PI3 K and AKT proteins(P＜0.01).After ASJYD treatment,the CUMS+ASJYD group exhibited significant improvements in sucrose consumption,preference rate,locomotor activity,and central zone exploration(P＜0.01),accompanied by reduced ACTH/CORT expression,alleviated hippocampal pathology,restored mitochondrial integrity with clear cristae,and increased PI3K/AKT phosphorylation(P＜0.01).Conclusion:Anshen Jieyu decoction significantly improved the depression-like be-haviour of CUMS rats,which may be achieved by down-regulating the hyperactive hypothalamic-pituitary-adrenal axis,regulating the expression of PI3K/AKT signaling pathway,and ameliorating the damage to hippocampal neurons and mi-tochondria.

Objective To study the effects of different extracorporeal circulation temperature combined with dexmedetomidine on oxidative stress in patients undergoing cardiac surgery under cardiopulmonary bypass.Methods 240 patients who underwent cardiac surgery under cardiopulmonary bypass were selected from June 2021 to June 2024.The patients were 50 to 75 years old and did not have severe hepatic or renal insufficiency.The Mini-Mental State Examination was completed 1 day before the operation.Patients were routinely established for cardiopulmonary bypass.Patients were randomly divided into four groups(n=55):hypothermia+normal saline group(L0),hypothermia+dexmedetomidine group(L1),hyperthermia+normal saline group(H0)and hyper-thermia+dexmedetomidine group(H1).The nasopharyngeal temperature was maintained at(30±1)℃in the hypothermia group and(33±1)℃in the high-temperature group during the reflux period.Dexmedetomidine injection was intravenously injected at 1 μg/kg 10 minutes before anesthesia in L1 and H1 groups,and pumped continuously at a rate of 0.5 μg/(kg·h)until the end of surgery.L0 and H0 groups were given equal volume of normal saline until the end of operation.5 mL of central venous blood was collected before the beginning of anesthesia(T1),at the end of surgery(T2),24 h(T3)and 48 h(T4)after surgery,and serum neuron specific enolase(NSE),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)levels were detected by ELISA.The levels of superoxide dismutase(SOD)and malondialdehyde(MDA)in serum were determined by xanthine oxidase method and thiobarbituric acid method.The Confusion Assessment Method-ICU was used to evaluate the occurrence of POD.Results 220 patients were finally enrolled,including 81 patients in POD group,with an incidence of 36.8%.Compared with NPOD group,the concentrations of NSE,IL-6,TNF-ɑ and MDA in POD group were in-creased,while the concentration of SOD was decreased.Compared with L0 group,the concentrations of NSE,IL-6,TNF-ɑ and MDA were decreased and the concentration of SOD increased in L1 group.Compared with H0 group,NSE,IL-6,TNF-ɑ and MDA concentrations in H1 group were decreased,while SOD concentration was in-creased.Compared with L0 group,concentration of NSE,IL-6,TNF-ɑ and MDA increased,while the concentra-tion of SOD decreased in H0 group.Compared with L1 group,concentration of NSE,IL-6,TNF-ɑ and MDA in-creased,while the concentration of SOD decreased in H1 group.Conclusion Hypothermia combined with dexme-detomidine during cardiopulmonary bypass can reduce oxidative stress response and POD in patients.

Objective To study the effects of different extracorporeal circulation temperature combined with dexmedetomidine on oxidative stress in patients undergoing cardiac surgery under cardiopulmonary bypass.Methods 240 patients who underwent cardiac surgery under cardiopulmonary bypass were selected from June 2021 to June 2024.The patients were 50 to 75 years old and did not have severe hepatic or renal insufficiency.The Mini-Mental State Examination was completed 1 day before the operation.Patients were routinely established for cardiopulmonary bypass.Patients were randomly divided into four groups(n=55):hypothermia+normal saline group(L0),hypothermia+dexmedetomidine group(L1),hyperthermia+normal saline group(H0)and hyper-thermia+dexmedetomidine group(H1).The nasopharyngeal temperature was maintained at(30±1)℃in the hypothermia group and(33±1)℃in the high-temperature group during the reflux period.Dexmedetomidine injection was intravenously injected at 1 μg/kg 10 minutes before anesthesia in L1 and H1 groups,and pumped continuously at a rate of 0.5 μg/(kg·h)until the end of surgery.L0 and H0 groups were given equal volume of normal saline until the end of operation.5 mL of central venous blood was collected before the beginning of anesthesia(T1),at the end of surgery(T2),24 h(T3)and 48 h(T4)after surgery,and serum neuron specific enolase(NSE),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)levels were detected by ELISA.The levels of superoxide dismutase(SOD)and malondialdehyde(MDA)in serum were determined by xanthine oxidase method and thiobarbituric acid method.The Confusion Assessment Method-ICU was used to evaluate the occurrence of POD.Results 220 patients were finally enrolled,including 81 patients in POD group,with an incidence of 36.8%.Compared with NPOD group,the concentrations of NSE,IL-6,TNF-ɑ and MDA in POD group were in-creased,while the concentration of SOD was decreased.Compared with L0 group,the concentrations of NSE,IL-6,TNF-ɑ and MDA were decreased and the concentration of SOD increased in L1 group.Compared with H0 group,NSE,IL-6,TNF-ɑ and MDA concentrations in H1 group were decreased,while SOD concentration was in-creased.Compared with L0 group,concentration of NSE,IL-6,TNF-ɑ and MDA increased,while the concentra-tion of SOD decreased in H0 group.Compared with L1 group,concentration of NSE,IL-6,TNF-ɑ and MDA in-creased,while the concentration of SOD decreased in H1 group.Conclusion Hypothermia combined with dexme-detomidine during cardiopulmonary bypass can reduce oxidative stress response and POD in patients.

Gut bacterial nitroreductases play an important role in reduction of various nitroaromatic compounds to the corresponding N-nitroso compounds, hydroxylamines or aromatic amines, most of which are carcinogenic and mutagenic agents. Inhibition of gut nitroreductases has been recognized as an attractive approach for reducing mutagen metabolites in the colon, so as to prevent colon diseases. In this study, the inhibitory effects of 55 herbal medicines against Escherichia coli(E. coli) nitroreductase (EcNfsA) were examined. Compared with other herbal extracts, Syzygium aromaticum extract showed superior inhibitory potency toward EcNfsA mediated nitrofurazone reduction. Then, the inhibitory effects of 22 major constituents in Syzygium aromaticum against EcNfsA were evaluted. Compared with other tested natural compounds, ellagic acid, corilagin, betulinic acid, oleanic acid, ursolic acid, urolithin M5 and isorhamnetin were found with strong to moderate inhibitory effect against EcNfsA, with IC₅₀ values ranging from 0.67 to 28.98 mol·L^-1. Furthermore, the inhibition kinetic analysis and docking simulation demonstrated that ellagic acid and betulinic acid potently inhibited EcNfsA (K_i < 2 μmol·L ^-1) in a competitively inhibitory manner, which created strong interactions with the catalytic triad of EcNfsA. In summary, our findings provide new scientific basis for explaining the anti-mutagenic activity of Syzygium aromaticum, where some newly identified EcNfsA inhibitors can be used for developing novel agents to reduce the toxicity induced by bacterial nitroreductase.
Ellagic Acid/pharmacology* ; Escherichia coli ; Kinetics ; Nitroreductases/pharmacology* ; Plant Extracts/pharmacology* ; Syzygium

Small proteins specifically refer to proteins consisting of less than 100 amino acids translated from small open reading frames (sORFs), which were usually missed in previous genome annotation. The significance of small proteins has been revealed in current years, along with the discovery of their diverse functions. However, systematic annotation of small proteins is still insufficient. SmProt was specially developed to provide valuable information on small proteins for scientific community. Here we present the update of SmProt, which emphasizes reliability of translated sORFs, genetic variants in translated sORFs, disease-specific sORF translation events or sequences, and remarkably increased data volume. More components such as non-ATG translation initiation, function, and new sources are also included. SmProt incorporated 638,958 unique small proteins curated from 3,165,229 primary records, which were computationally predicted from 419 ribosome profiling (Ribo-seq) datasets or collected from literature and other sources from 370 cell lines or tissues in 8 species (Homo sapiens, Mus musculus, Rattus norvegicus, Drosophila melanogaster, Danio rerio, Saccharomyces cere-visiae, Caenorhabditis elegans, and Escherichia coli). In addition, small protein families identified from human micro-biomes were also collected. All datasets in SmProt are free to access, and available for browse, search, and bulk downloads at http://bigdata.ibp.ac.cn/SmProt/.

Pancreatic lipase (PL), a crucial enzyme in the digestive system of mammals, has been proven as a therapeutic target to prevent and treat obesity. The purpose of this study is to evaluate and characterize the PL inhibition activities of the major constituents from Fructus Psoraleae (FP), one of the most frequently used Chinese herbs with lipid-lowering activity. To this end, a total of eleven major constituents isolated from Fructus Psoraleae have been obtained and their inhibition potentials against PL have been assayed by a fluorescence-based assay. Among all tested compounds, isobavachalcone, bavachalcone and corylifol A displayed strong inhibition on PL (IC < 10 μmol·L). Inhibition kinetic analyses demonstrated that isobavachalcone, bavachalcone and corylifol A acted as mixed inhibitors against PL-mediated 4-methylumbelliferyl oleate (4-MUO) hydrolysis, with the K values of 1.61, 3.77 and 10.16 μmol·L, respectively. Furthermore, docking simulations indicated that two chalcones (isobavachalcone and bavachalcone) could interact with the key residues located in the catalytic cavity of PL via hydrogen binding and hydrophobic interactions. Collectively, these finding provided solid evidence to support that Fructus Psoraleae contained bioactive compounds with lipid-lowering effects via targeting PL, and also suggested that the chalcones in Fructus Psoraleae could be used as ideal leading compounds to develop novel PL inhibitors.

Objective To investigate the application of intraoperative neuromonitoring (IONM) during thyroidectomy for external branch of superior laryngeal nerve(EBSLN).Methods From Jan.2017 to Jun.2017,138 patients undergoing thyroidectomy were randomly divided into monitor group (n=69) and the control group (n=69).The monitor group were used IONM for EBSLN,while the control group were used conventional area protection.Results The overall incidence of EBSLN injury was 1.4％(1/69) in the monitor group,and the overall incidence of EBSLN injury was 11.6％(8/69) in the control group.There was statistical significance between the two groups.Conclusion The application of IONM in thyroidectomy can exactly identify EBSLN,and reduce the possibility of EBSLN injury remarkably.

Aberrant regulation of miRNA genes contributes to pathogenesis of a wide range of human diseases, including cancer. The TAR DNA binding protein 43 (TDP-43), a RNA/DNA binding protein associated with neurodegeneration, is involved in miRNA biogenesis. Here, we systematically examined miRNAs regulated by TDP-43 using RNA-Seq coupled with an siRNA-mediated knockdown approach. TDP-43 knockdown affected the expression of a number of miRNAs. In addition, TDP-43 down-regulation led to alterations in the patterns of different isoforms of miRNAs (isomiRs) and miRNA arm selection, suggesting a previously unknown role of TDP-43 in miRNA processing. A number of TDP-43 associated miRNAs, and their candidate target genes, are associated with human cancers. Our data reveal highly complex roles of TDP-43 in regulating different miRNAs and their target genes. Our results suggest that TDP-43 may promote migration of lung cancer cells by regulating miR-423-3p. In contrast, TDP-43 increases miR-500a-3p expression and binds to the mature miR-500a-3p sequence. Reduced expression of miR-500a-3p is associated with poor survival of lung cancer patients, suggesting that TDP-43 may have a suppressive role in cancer by regulating miR-500a-3p. Cancer-associated genes LIF and PAPPA are possible targets of miR-500a-3p. Our work suggests that TDP-43-regulated miRNAs may play multifaceted roles in the pathogenesis of cancer.
Animals ; Cells, Cultured ; DNA-Binding Proteins ; metabolism ; Electrophoretic Mobility Shift Assay ; Humans ; Immunoprecipitation ; Mice ; MicroRNAs ; genetics ; metabolism ; Neoplasms ; genetics ; metabolism