Background:Drug-induced liver damage is a severe medical issue that affects people all over the world.Sorafenib has some side effects that cause liver injury.A dietary medicinal plant called Penthorum chinense Pursh.(...Background:Drug-induced liver damage is a severe medical issue that affects people all over the world.Sorafenib has some side effects that cause liver injury.A dietary medicinal plant called Penthorum chinense Pursh.(PCP)has hepatoprotective properties.There are currently few reports on PCP’s protective impact and mechanism against sorafenib-induced liver injury.Methods:To create a liver injury model,sorafenib was administered to BRL-3A cells.Cell viability assays,immunofluorescence tests,Western blotting,real-time quantitative PCR,and high-content imaging systems were utilized to examine PCP’s effect and mechanism.Results:In this study,PCP treatment mitigated the liver damage caused by sorafenib by enhancing cell survival,lowering lipid reactive oxygen species and malondialdehyde levels,and elevating glutathione levels.In addition,PCP can enhance the protein expression of cystine/glutamate transporter xCT and glutathione peroxidase 4,reduce iron content and alleviate mitochondrial toxicity.Further mechanism studies revealed that PCP inhibited ferroptosis by promoting the production of nuclear factor E2-related factor 2 nuclear translocation and subsequently affecting target genes(HO-1 and NQO1).Conclusion:Together,PCP regulates the nuclear factor E2-related factor 2 pathway,which helps to lessen ferroptosis brought on by sorafenib.展开更多
Background:Luteolin is a flavonoid chemical that exists in a variety of medicinal and edible plants and holds many biologically active properties in liver protection,anti-cancer,antioxidants,anti-inflammatory,neuropro...Background:Luteolin is a flavonoid chemical that exists in a variety of medicinal and edible plants and holds many biologically active properties in liver protection,anti-cancer,antioxidants,anti-inflammatory,neuroprotective,etc.According to its hepatoprotective properties,luteolin was selected to co-treat with sorafenib,one of the approved protein kinase inhibitors,to reduce sorafenib-induced normal liver cell damage.Methods:The BRL-3A cell line was treated with sorafenib to establish a liver injury model,followed by luteolin treatment.The cell viability was detected,and the mechanism of action was detected by immunofluorescence,western blotting,and real-time quantitative PCR.Results:The research findings demonstrated that luteolin could increase cystine/glutamate transporter xCT(SLC7A11)and glutathione peroxidase 4(GPX4)expression and display a chelating effect on iron,which led to increased glutathione and decreased malondialdehyde,Fe^(2+) and lipid reactive oxygen species contents in BRL-3A cells,and the sorafenib-induced mitochondrial membrane potential decrease was also inhibited.In addition,when sorafenib caused the accumulation of lipid reactive oxygen species,luteolin could help release this oxidative stress by activating nuclear factor E2-related factor 2(Nrf2)and up-regulating the expression of the associated genes heme oxygenase 1(HO-1)and quinone oxidoreductase 1(NQO1).Conclusion:Therefore,luteolin may ameliorate sorafenib-induced ferroptosis by activating the Nrf2-associated pathway without any impact on sorafenib anti-cancer activity.It can be used as an adjuvant to sorafenib to reduce liver injury in patients with hepatocellular carcinoma.展开更多
AIM To investigate the influence of high salt on dextran sulfate sodium(DSS)-induced colitis in mice and explore the underlying mechanisms of this effect.METHODS DSS and NaC l were used to establish the proinflammator...AIM To investigate the influence of high salt on dextran sulfate sodium(DSS)-induced colitis in mice and explore the underlying mechanisms of this effect.METHODS DSS and NaC l were used to establish the proinflammatory animal model. We evaluated the colitis severity. Flow cytometry was employed for detecting the frequencies of Th1, macrophages and Tregs in spleen, mesenteric lymph node and lamina propria. The important role of macrophages in the promotion of DSS-induced colitis by NaCl was evaluated by depleting macrophages with clodronate liposomes. Activated peritoneal macrophages and lamina propria mononuclear cells(LPMCs) were stimulated with NaCl, and proteins were detected by western blotting. Cytokines and inflammation genes were analyzed by enzyme-linked immunosorbent assay and RT-PCR, respectively.RESULTS The study findings indicate that NaC l up-regulates the frequencies of CD11b^+ macrophages and CD4^+IFN-γ^+IL-17^+ T cells in lamina propria in DSS-treated mice. CD3^+CD4^+CD25^+Foxp^3+ T cells, which can secrete high levels of IL-10 and TGF-β, increase through feedback in NaCl-and DSS-treated mice. Furthermore, clodronate liposomes pretreatment significantly alleviated DSSinduced colitis, indicating that macrophages play a vital role in NaCl proinflammatory activity. NaCl aggravates peritoneal macrophage inflammation by promoting the expressions of interleukin(IL)-1, IL-6 and mouse inducible nitric oxide synthase. Specifically, high NaCl concentrations promote p38 phosphorylation in lipopolysaccharide-and IFN-γ-activated LPMCs mediated by SGK1. CONCLUSION Proinflammatory macrophages may play an essential role in the onset and development of NaCl-promoted inflammation in DSS-induced colitis. The underlining mechanism involves up-regulation of the p38/MAPK axis.展开更多
To determine the molecular mechanism of cerebral ischemia/reperfusion injury, we examined the micro RNA（mi RNA） expression profile in rat cortex after focal cerebral ischemia/reperfusion injury using mi RNA microarr...To determine the molecular mechanism of cerebral ischemia/reperfusion injury, we examined the micro RNA（mi RNA） expression profile in rat cortex after focal cerebral ischemia/reperfusion injury using mi RNA microarrays and bioinformatic tools to systematically analyze Gene Ontology（GO） function classifications, as well as the signaling pathways of genes targeted by these differentially expressed mi RNAs. Our results show significantly changed mi RNA expression profiles in the reperfusion period after focal cerebral ischemia, with a total of 15 mi RNAs up-regulated and 44 mi RNAs down-regulated. Target genes of these differentially expressed mi RNAs were mainly involved in metabolic and cellular processes, which were identified as hub nodes of a mi RNA-GO-network. The most correlated pathways included D-glutamine and D-glutamate metabolism, the renin-angiotensin system, peroxisomes, the PPAR signaling pathway, SNARE interactions in vesicular transport, and the calcium signaling pathway. Our study suggests that mi RNAs play an important role in the pathological process of cerebral ischemia/reperfusion injury. Understanding mi RNA expression and function may shed light on the molecular mechanism of cerebral ischemia/reperfusion injury.展开更多
AIM:To detect human papillomavirus(HPV) DNA in esophageal carcinoma(EC) 109 cells and investigate the relationship between HPV and EC.METHODS:Genomic DNA and total RNA from EC109 cells were isolated.HPV DNA was detect...AIM:To detect human papillomavirus(HPV) DNA in esophageal carcinoma(EC) 109 cells and investigate the relationship between HPV and EC.METHODS:Genomic DNA and total RNA from EC109 cells were isolated.HPV DNA was detected by polymerase chain reaction(PCR) with the general primer sets of My09/11 and GP5 +/6 + for the HPV L1 gene and type-specific primer sets for HPV18 E6 and HPV18 E6-E7.Reverse transcription(RT) of mRNA isolated from EC109 cells was performed to produce a cDNA.And then a PCR-based protocol for the amplification of papillomavirus oncogene transcripts was used to analyze HPV18 DNA and integrated transcripts of HPV18 in the chromosomes of EC109 cells.The final nested PCR products were cloned into a pMD-18T vector and sequenced to analyze the chromosomal location of HPV integration.RESULTS:HPV18 DNA was detected in EC109 cells by PCR using the general primer sets of My09/11 and GP5 +/6 + for HPV L1 and the type-specif ic primer sets for HPV18 E6 and E6-E7 to generate products of 450 bp,150 bp,335 bp and 944 bp,respectively.Approximately 600 bp of integrated HPV18-specific transcript was identified.The final nested PCR product of integrated HPV18 DNA was cloned into a pMD-18T vector and sequenced to analyze the chromosomal location of HPV integration.Sequence alignment showed that the HPV18 sequence from EC109 cells was identical to that of the encoded early protein E7-E1 of the standard HPV18 strain X05015,and another partial gene sequence was identical to a partial sequence of human chromosome 8.CONCLUSION:Integration of the HPV genome into the host cell chromosome suggests that persistent HPV infection is vital for malignant cell transformation and carcinogenesis.展开更多
AIM: to establish a new animal model for the research of human rotavirus(HRV) infection, its pathogenesis and immunity and evaluation of potential vaccines.METHODS: 5-d, 30-d and 60-d-old Chinese mini-pigs, Guizhou an...AIM: to establish a new animal model for the research of human rotavirus(HRV) infection, its pathogenesis and immunity and evaluation of potential vaccines.METHODS: 5-d, 30-d and 60-d-old Chinese mini-pigs, Guizhou and bamma, were inoculated with a single oral dose of attenuated strain Wa, G1, G3 of HRV, and PbS(control), respectively, and fecal samples of pigs from 0 to 7 d post infection(DPI) were collected individually. Enzyme linked immunosorbent assay was used to detect HRV antigen in feces. the HRV was tested by real-time PCR(Rt-PCR). the sections of the intestinal tissue were stained with hematoxylin and eosin to observe the morphologic variation by microscopy. Immunofluorescence was used to determine the HRV in intestinal tissue. HRV particles in cells of the ileum were observed by electron micrography.RESULTS: When inoculated with HRV, mini-pigs younger than 30 d developed diarrhea in an agedependent manner and shed HRV antigen of the sameinoculum, as demonstrated by Rt- PCR.Histopathological changes were observed in HRV inoculated mini-pigs including small intestinal cell tumefaction and necrosis. HRV that was distributed in the small intestine was restricted to the top part of the villi on the internal wall of the ileum, which was observed by immunofluorescence and transmission electron microscopy. Virus particles were observed in Golgi like follicles in HRV-infected neonatal minipigs. Guizhou mini-pigs were more sensitive to HRV than bamma with respect to RV antigen shedding and clinical diarrhea.CONCLUSION: these results indicate that we have established a mini-pig model of HRV induced diarrhea. Our findings are useful for the understanding of the pathogenic mechanisms of HRV infection.展开更多
AIM:to study the prevalence of human papillomavirus(HPV) in esophageal carcinoma in tangshan,China,a high-incidence area.METHODS:Formalin-fixed,paraffin-embedded tissue specimens from 198 patients who were pathologica...AIM:to study the prevalence of human papillomavirus(HPV) in esophageal carcinoma in tangshan,China,a high-incidence area.METHODS:Formalin-fixed,paraffin-embedded tissue specimens from 198 patients who were pathologically diagnosed with esophageal squamous cell carcinoma from 2011 to 2013 were obtained from a pathology department in Tangshan.DNA was extracted from all198 specimens to detect HPV by polymerase chain reaction(PCR).β-globin PCR was performed to check the quality of the DNA extraction procedure.PCR was performed to detect a wide range of HPV types,and type-specific PCR was performed to detect HPV types16 and 18.Negative and positive controls were used for HPV 16 and 18 detection.RESULTS:the DNA extraction method in this study appeared to be more effective than other previously reported methods.After DNA extraction,more than98%of the tissue specimens had an acceptable result in the DNA qualification test(β-globin PCR).the overall prevalence of HPV in tumor tissues by GP6+/GP5+PCR was 79.79%,and the prevalence of HPV types16 and 18 was 40.40%and 47.47%,respectively.PCR demonstrated the presence of HPV,and direct sequencing confirmed the HPV genotypes.All HPVpositive PCR products were checked by DNA sequence analysis using DNAman and compared with the known HPV sequences listed in the basic Local Alignment Search tool database to evaluate the HPV types.this analysis confirmed the presence of HPV types 16 and18.CONCLUSION:DNA of high-risk HPV types 16 and 18is present in esophageal tumors,implicating HPV as a possible etiologic factor for esophageal squamous cell carcinoma.展开更多
Correction to:Signal Transduction and Targeted Therapy https://doi.org/10.1038/s41392-022-01017-8,published online 22 June 2022 In the process of collating the raw data,the authors noticed one inadvertent mistake occu...Correction to:Signal Transduction and Targeted Therapy https://doi.org/10.1038/s41392-022-01017-8,published online 22 June 2022 In the process of collating the raw data,the authors noticed one inadvertent mistake occurred in Fig.2e that needs to be corrected in the article1.The correct data are provided as follows.The key findings of the article are not affected by the correction.The original article has been corrected.展开更多
Objective:To explore whether the ethanol extract of Herpetospermum caudigerum Wall(EHC),a Xizang medicinal plant traditionally used for treating liver diseases,can improve imiquimod-induced psoriasis-like skin inflamm...Objective:To explore whether the ethanol extract of Herpetospermum caudigerum Wall(EHC),a Xizang medicinal plant traditionally used for treating liver diseases,can improve imiquimod-induced psoriasis-like skin inflammation.Methods:Immunohistochemistry and immunofluorescence staining were used to determine the effects of topical EHC use in vivo on the skin pathology of imiquimod-induced psoriasis in mice.The protein levels of interferon-γ(IFN-γ),tumor necrosis factor-a(TNF-a),and interleukin-17A(IL-17A)in mouse skin samples were examined using immunohistochemical staining.In vitro,IFN-γ-induced HaCaT cells with or without EHC treatment were used to evaluate the expression of keratinocyte-derived intercellular cell adhesion molecule-1(ICAM-1)and chemokine CXC ligand 9(CXCL9)using Western blotting and reverse transcription-quantitative polymerase chain reaction.The protein synthesis inhibitor cycloheximide and proteasome inhibitor MG132 were utilized to validate the EHC-mediated mechanism underlying degradation of ICAM-1 and CXCL9.Results:EHC improved inflammation in the imiquimod-induced psoriasis mouse model and reduced the levels of IFN-γ,TNF-a,and IL-17A in psoriatic lesions.Treatment with EHC also suppressed ICAM-1 and CXCL9 in epidermal keratinocytes.Further mechanistic studies revealed that EHC suppressed keratinocyte-derived ICAM-1 and CXCL9 by promoting ubiquitin–proteasome-mediated protein degradation rather than transcriptional repression.Seven primary compounds including ehletianol C,dehydrodiconiferyl alcohol,herpetrione,herpetin,herpetotriol,herpetetrone and herpetetrol were identified from the EHC using ultra-performance liquid chromatography-quadrupole-time of flight-mass spectrometry.Conclusion:Topical application of EHC ameliorates psoriasis-like skin symptoms and improves the inflammation at the lesion sites.展开更多
Platelet-derived growth factor receptor alpha （PDGFRct） is a marker of oligodendrocyte precursor cells in the central nervous system. NG2 is also considered a marker of oligodendrocyte precursor cells. However, whet...Platelet-derived growth factor receptor alpha （PDGFRct） is a marker of oligodendrocyte precursor cells in the central nervous system. NG2 is also considered a marker of oligodendrocyte precursor cells. However, whether there are differences in the distribution and morphol- ogy of oligodendrocyte precursor cells labeled by NG2 or PDGFRa in the developing neonatal rat brain remains unclear. In this study, by immunohistochemical staining, NG2 positive （NG2＋） cells were ubiquitous in the molecular layer, external pyramidal layer, internal pyramidal layer, and polymorphic layer of the cerebral cortex, and corpus callosum, external capsule, piriform cortex, and medial septal nucleus. NG2~ cells were stellate or fusiform in shape with long processes that were progressively decreased and shortened over the course of brain development. The distribution and morphology of PDGFRct positive （PDGFRa＋） cells were coincident with NG2＋ cells. The co- localization of NG2 and PDGFRu in the cell bodies and processes of some cells was confirmed by double immunofluorescence labeling. Moreover, cells double-labeled for NG2 and PDGFRa were predominantly in the early postnatal stage of development. The numbers of NG2＋/PDGFRa＋ cells and PDGFRa＋ cells decreased, but the number of NG2＋ cells increased from postnatal days 3 to 14 in the developing brain. In addition, amoeboid microglial cells of the corpus callosum, newborn brain macrophages in the normal developing brain, did not express NG2 or PDGFRu, but NG2 expression was detected in amoeboid microglia after hypoxia. The present results suggest that NG2 and PDGFRct are specific markers of oligodendrocyte precursor cells at different stages during early development. Additionally, the NG2 protein is involved in inflammatory and pathological processes of amoeboid microglial cells.展开更多
Pancreatic ductal adenocarcinoma(PDAC)is well-known for inefficient early diagnosis,with most patients diagnosed at advanced stages.Increasing evidence indicates that elevated plasma levels of branched-chain amino aci...Pancreatic ductal adenocarcinoma(PDAC)is well-known for inefficient early diagnosis,with most patients diagnosed at advanced stages.Increasing evidence indicates that elevated plasma levels of branched-chain amino acids(BCAAs)are associated with an increased risk of pancreatic cancer.Branched-chain amino acid transaminase 2(BCAT2)is an important enzyme in BCAA catabolism that reversibly catalyzes the initial step of BCAA degradation to branched-chain acyl-CoA.Here,we show that BCAT2 is acetylated at lysine 44(K44),an evolutionarily conserved residue.BCAT2 acetylation leads to its degradation through the ubiquitin–proteasome pathway and is stimulated in response to BCAA deprivation.cAMP-responsive element-binding(CREB)-binding protein(CBP)and SIRT4 are the acetyltransferase and deacetylase for BCAT2,respectively.CBP and SIRT4 bind to BCAT2 and control the K44 acetylation level in response to BCAA availability.More importantly,the K44R mutant promotes BCAA catabolism,cell proliferation,and pancreatic tumor growth.Collectively,the data from our study reveal a previously unknown regulatory mechanism of BCAT2 in PDAC and provide a potential therapeutic target for PDAC treatment.展开更多
Folic acid,served as dietary supplement,is closely linked to one-carbon metabolism and methionine metabolism.Previous clinical evidence indicated that folic acid supplementation displays dual effect on cancer developm...Folic acid,served as dietary supplement,is closely linked to one-carbon metabolism and methionine metabolism.Previous clinical evidence indicated that folic acid supplementation displays dual effect on cancer development,promoting or suppressing tumor formation and progression.However,the underlying mechanism remains to be uncovered.Here,we report that high-folate diet significantly promotes cancer development in mice with hepatocellular carcinoma(HCC)induced by DEN/high-fat diet(HFD),simultaneously with increased expression of methionine adenosyltransferase 2A(gene name,MAT2A;protein name,MATIIα),the key enzyme in methionine metabolism,and acceleration of methionine cycle in cancer tissues.In contrast,folate-free diet reduces MATIIαexpression and impedes HFD-induced HCC development.Notably,methionine metabolism is dynamically reprogrammed with valosin-containing protein p97/p47 complex-interacting protein(VCIP135)which functions as a deubiquitylating enzyme to bind and stabilize MATIIαin response to folic acid signal.Consistently,upregulation of MATIIαexpression is positively correlated with increased VCIP135 protein level in human HCC tissues compared to adjacent tissues.Furthermore,liver-specific knockout of Mat2a remarkably abolishes the advocating effect of folic acid on HFD-induced HCC,demonstrating that the effect of high or free folate-diet on HFD-induced HCC relies on Mat2a.Moreover,folate and multiple intermediate metabolites in one-carbon metabolism are significantly decreased in vivo and in vitro upon Mat2a deletion.Together,folate promotes the integration of methionine and one-carbon metabolism,contributing to HCC development via hijacking MATIIαmetabolic pathway.This study provides insight into folate-promoted cancer development,strongly recommending the tailor-made folate supplement guideline for both sub-healthy populations and patients with cancer expressing high level of MATIIαexpression.展开更多
Background:The shortage of donor corneas is a severe global issue,and hence the development of corneal alternatives is imperative and urgent.Although attempts to produce artificial cornea substitutes by tissue enginee...Background:The shortage of donor corneas is a severe global issue,and hence the development of corneal alternatives is imperative and urgent.Although attempts to produce artificial cornea substitutes by tissue engineering have made some positive progress,many problems remain that hamper their clinical application worldwide.For example,the curvature of tissue-engineered cornea substitutes cannot be designed to fit the bulbus oculi of patients.Objective:To overcome these limitations,in this paper,we present a novel integrated three-dimensional(3 D) bioprintingbased cornea substitute fabrication strategy to realize design,customized fabrication,and evaluation of multi-layer hollow structures with complicated surfaces.Methods:The key rationale for this method is to combine digital light processing(DLP) and extrusion bioprinting into an integrated 3 D cornea bioprinting system.A designable and personalized corneal substitute was designed based on mathematical modelling and a computer tomography scan of a natural cornea.The printed corneal substitute was evaluated based on biomechanical analysis,weight,structural integrity,and fit.Results:The results revealed that the fabrication of high water content and highly transparent curved films with geometric features designed according to the natural human cornea can be achieved using a rapid,simple,and low-cost manufacturing process with a high repetition rate and quality.Conclusions:This study demonstrated the feasibility of customized design,analysis,and fabrication of a corneal substitute.The programmability of this method opens up the possibility of producing substitutes for other cornea-like shell structures with different scale and geometry features,such as the glomerulus,atrium,and oophoron.展开更多
Epithelial ovarian cancer(EOC) exhibits strong dependency on the tricarboxylic acid(TCA) cycle and oxidative phosphorylation to fuel anabolic process.Here,we show that malate dehydrogenase 2(MDH2),a key enzyme of the ...Epithelial ovarian cancer(EOC) exhibits strong dependency on the tricarboxylic acid(TCA) cycle and oxidative phosphorylation to fuel anabolic process.Here,we show that malate dehydrogenase 2(MDH2),a key enzyme of the TCA cycle,is palmitoylated at cysteine 138(C138) residue,resulting in increased activity of MDH2.We next identify that ZDHHC18 acts as a palmitoyltransferase of MDH2.Glutamine deprivation enhances MDH2 palmitoylation by increasing the binding between ZDHHC18 and MDH2.MDH2 silencing represses mitochondrial respiration as well as ovarian cancer cell proliferation both in vitro and in vivo.Intriguingly,re-expression of wild-type MDH2,but not its palmitoylation-deficient C138 S mutant,sustains mitochondrial respiration and restores the growth as well as clonogenic capability of ovarian cancer cells.Notably,MDH2 palmitoylation level is elevated in clinical cancer samples from patients with high-grade serous ovarian cancer.These observations suggest that MDH2 palmitoylation catalyzed by ZDHHC18 sustains mitochondrial respiration and promotes the malignancy of ovarian cancer,yielding possibilities of targeting ZDHHC18-mediated MDH2 palmitoylation in the treatment of EOC.展开更多
Electromagnetic field is an available online method to increase bonding strength of clad sheet manufactured by horizontal twin-roll casting （HTRC）. In this paper, an electric current pulse （ECP） and a complex fie...Electromagnetic field is an available online method to increase bonding strength of clad sheet manufactured by horizontal twin-roll casting （HTRC）. In this paper, an electric current pulse （ECP） and a complex field （static magnetic field （SMF） together with ECP） are exerted during HTRC of steel/aluminum clad sheet. The produced clad sheet has good appearance, and no visible defects exist at the bonding interface. The inter-diffusion zone at Fe/A1 interface in ECP and SMF＋ECP sheets is 3 and 4 μm, respectively, and the latter increases slightly compared with that in non-field sheet. The average peel strengths （APS） of ECP and SMF＋ECP sheet are 14 and 21 N/mm, respectively, which increase by 2 and 9 N/mm compared with 12 N/mm of non-field sheet. The APS increment in SMF＋ECP sheet is resulted from the increment of interface bonding spots and the enhancement of inter-diffusion zone width.展开更多
To the Editor： Laparoendoscopic single-site surgery （LESS） has been widely used for various urologic diseases in order to improve safety, outcomes, and cosmesis, and to further reduce invasiveness. Here, we reporte...To the Editor： Laparoendoscopic single-site surgery （LESS） has been widely used for various urologic diseases in order to improve safety, outcomes, and cosmesis, and to further reduce invasiveness. Here, we reported our initial experience with laparoendoscopic single-site retroperitoneal partial adrenalectomy using a homemade single-port device in a municipal hospital.展开更多
基金supported by the open fund of State Key Laboratory of Southwestern Chinese Medicine Resources(No.SCMR202103)to Jian LiTibet Autonomous Region Science and Technology Plan(high-tech social development)project(No.XZ202201ZY0031G)to Yi-Xi YangAnti-infective Agent Creation Engineering Research Centre of Sichuan Province,Sichuan Industrial Institute of Antibiotics,School of pharmacy,Chengdu University(No.AAC2023002)to Qiu-Xia Lu.
文摘Background:Drug-induced liver damage is a severe medical issue that affects people all over the world.Sorafenib has some side effects that cause liver injury.A dietary medicinal plant called Penthorum chinense Pursh.(PCP)has hepatoprotective properties.There are currently few reports on PCP’s protective impact and mechanism against sorafenib-induced liver injury.Methods:To create a liver injury model,sorafenib was administered to BRL-3A cells.Cell viability assays,immunofluorescence tests,Western blotting,real-time quantitative PCR,and high-content imaging systems were utilized to examine PCP’s effect and mechanism.Results:In this study,PCP treatment mitigated the liver damage caused by sorafenib by enhancing cell survival,lowering lipid reactive oxygen species and malondialdehyde levels,and elevating glutathione levels.In addition,PCP can enhance the protein expression of cystine/glutamate transporter xCT and glutathione peroxidase 4,reduce iron content and alleviate mitochondrial toxicity.Further mechanism studies revealed that PCP inhibited ferroptosis by promoting the production of nuclear factor E2-related factor 2 nuclear translocation and subsequently affecting target genes(HO-1 and NQO1).Conclusion:Together,PCP regulates the nuclear factor E2-related factor 2 pathway,which helps to lessen ferroptosis brought on by sorafenib.
基金supported by the open fund of State Key Laboratory of Southwestern Chinese Medicine Resources(No.SCMR202103)to Jian LiTibet Autonomous Region Science and Technology Plan(high-tech social development)project(No.XZ202201ZY0031G)to Yang YXAnti-infective Agent Creation Engineering Research Centre of Sichuan Province,Sichuan Industrial Institute of Antibiotics,School of pharmacy,Chengdu University(No.AAC2023002)to Lu QX.
文摘Background:Luteolin is a flavonoid chemical that exists in a variety of medicinal and edible plants and holds many biologically active properties in liver protection,anti-cancer,antioxidants,anti-inflammatory,neuroprotective,etc.According to its hepatoprotective properties,luteolin was selected to co-treat with sorafenib,one of the approved protein kinase inhibitors,to reduce sorafenib-induced normal liver cell damage.Methods:The BRL-3A cell line was treated with sorafenib to establish a liver injury model,followed by luteolin treatment.The cell viability was detected,and the mechanism of action was detected by immunofluorescence,western blotting,and real-time quantitative PCR.Results:The research findings demonstrated that luteolin could increase cystine/glutamate transporter xCT(SLC7A11)and glutathione peroxidase 4(GPX4)expression and display a chelating effect on iron,which led to increased glutathione and decreased malondialdehyde,Fe^(2+) and lipid reactive oxygen species contents in BRL-3A cells,and the sorafenib-induced mitochondrial membrane potential decrease was also inhibited.In addition,when sorafenib caused the accumulation of lipid reactive oxygen species,luteolin could help release this oxidative stress by activating nuclear factor E2-related factor 2(Nrf2)and up-regulating the expression of the associated genes heme oxygenase 1(HO-1)and quinone oxidoreductase 1(NQO1).Conclusion:Therefore,luteolin may ameliorate sorafenib-induced ferroptosis by activating the Nrf2-associated pathway without any impact on sorafenib anti-cancer activity.It can be used as an adjuvant to sorafenib to reduce liver injury in patients with hepatocellular carcinoma.
基金Supported by National Natural Science Foundation of China,No.81271813 and No.81570497
文摘AIM To investigate the influence of high salt on dextran sulfate sodium(DSS)-induced colitis in mice and explore the underlying mechanisms of this effect.METHODS DSS and NaC l were used to establish the proinflammatory animal model. We evaluated the colitis severity. Flow cytometry was employed for detecting the frequencies of Th1, macrophages and Tregs in spleen, mesenteric lymph node and lamina propria. The important role of macrophages in the promotion of DSS-induced colitis by NaCl was evaluated by depleting macrophages with clodronate liposomes. Activated peritoneal macrophages and lamina propria mononuclear cells(LPMCs) were stimulated with NaCl, and proteins were detected by western blotting. Cytokines and inflammation genes were analyzed by enzyme-linked immunosorbent assay and RT-PCR, respectively.RESULTS The study findings indicate that NaC l up-regulates the frequencies of CD11b^+ macrophages and CD4^+IFN-γ^+IL-17^+ T cells in lamina propria in DSS-treated mice. CD3^+CD4^+CD25^+Foxp^3+ T cells, which can secrete high levels of IL-10 and TGF-β, increase through feedback in NaCl-and DSS-treated mice. Furthermore, clodronate liposomes pretreatment significantly alleviated DSSinduced colitis, indicating that macrophages play a vital role in NaCl proinflammatory activity. NaCl aggravates peritoneal macrophage inflammation by promoting the expressions of interleukin(IL)-1, IL-6 and mouse inducible nitric oxide synthase. Specifically, high NaCl concentrations promote p38 phosphorylation in lipopolysaccharide-and IFN-γ-activated LPMCs mediated by SGK1. CONCLUSION Proinflammatory macrophages may play an essential role in the onset and development of NaCl-promoted inflammation in DSS-induced colitis. The underlining mechanism involves up-regulation of the p38/MAPK axis.
基金supported by grants from the National Natural Science Foundation of ChinaNo.81271358+1 种基金Yunnan Science Foundation of ChinaNo.2013FZ199
文摘To determine the molecular mechanism of cerebral ischemia/reperfusion injury, we examined the micro RNA（mi RNA） expression profile in rat cortex after focal cerebral ischemia/reperfusion injury using mi RNA microarrays and bioinformatic tools to systematically analyze Gene Ontology（GO） function classifications, as well as the signaling pathways of genes targeted by these differentially expressed mi RNAs. Our results show significantly changed mi RNA expression profiles in the reperfusion period after focal cerebral ischemia, with a total of 15 mi RNAs up-regulated and 44 mi RNAs down-regulated. Target genes of these differentially expressed mi RNAs were mainly involved in metabolic and cellular processes, which were identified as hub nodes of a mi RNA-GO-network. The most correlated pathways included D-glutamine and D-glutamate metabolism, the renin-angiotensin system, peroxisomes, the PPAR signaling pathway, SNARE interactions in vesicular transport, and the calcium signaling pathway. Our study suggests that mi RNAs play an important role in the pathological process of cerebral ischemia/reperfusion injury. Understanding mi RNA expression and function may shed light on the molecular mechanism of cerebral ischemia/reperfusion injury.
基金Supported by An independent research fund from the National Institute for Viral Disease Control and Prevention,the Chinese Center for Disease Control and Preventionthe State Key Laboratory for Infectious Disease Prevention and Control (Grant No. 2011SKLID103)
文摘AIM:To detect human papillomavirus(HPV) DNA in esophageal carcinoma(EC) 109 cells and investigate the relationship between HPV and EC.METHODS:Genomic DNA and total RNA from EC109 cells were isolated.HPV DNA was detected by polymerase chain reaction(PCR) with the general primer sets of My09/11 and GP5 +/6 + for the HPV L1 gene and type-specific primer sets for HPV18 E6 and HPV18 E6-E7.Reverse transcription(RT) of mRNA isolated from EC109 cells was performed to produce a cDNA.And then a PCR-based protocol for the amplification of papillomavirus oncogene transcripts was used to analyze HPV18 DNA and integrated transcripts of HPV18 in the chromosomes of EC109 cells.The final nested PCR products were cloned into a pMD-18T vector and sequenced to analyze the chromosomal location of HPV integration.RESULTS:HPV18 DNA was detected in EC109 cells by PCR using the general primer sets of My09/11 and GP5 +/6 + for HPV L1 and the type-specif ic primer sets for HPV18 E6 and E6-E7 to generate products of 450 bp,150 bp,335 bp and 944 bp,respectively.Approximately 600 bp of integrated HPV18-specific transcript was identified.The final nested PCR product of integrated HPV18 DNA was cloned into a pMD-18T vector and sequenced to analyze the chromosomal location of HPV integration.Sequence alignment showed that the HPV18 sequence from EC109 cells was identical to that of the encoded early protein E7-E1 of the standard HPV18 strain X05015,and another partial gene sequence was identical to a partial sequence of human chromosome 8.CONCLUSION:Integration of the HPV genome into the host cell chromosome suggests that persistent HPV infection is vital for malignant cell transformation and carcinogenesis.
基金Supported by National Natural Science Foundation of China,No.30400402 and No.30571708
文摘AIM: to establish a new animal model for the research of human rotavirus(HRV) infection, its pathogenesis and immunity and evaluation of potential vaccines.METHODS: 5-d, 30-d and 60-d-old Chinese mini-pigs, Guizhou and bamma, were inoculated with a single oral dose of attenuated strain Wa, G1, G3 of HRV, and PbS(control), respectively, and fecal samples of pigs from 0 to 7 d post infection(DPI) were collected individually. Enzyme linked immunosorbent assay was used to detect HRV antigen in feces. the HRV was tested by real-time PCR(Rt-PCR). the sections of the intestinal tissue were stained with hematoxylin and eosin to observe the morphologic variation by microscopy. Immunofluorescence was used to determine the HRV in intestinal tissue. HRV particles in cells of the ileum were observed by electron micrography.RESULTS: When inoculated with HRV, mini-pigs younger than 30 d developed diarrhea in an agedependent manner and shed HRV antigen of the sameinoculum, as demonstrated by Rt- PCR.Histopathological changes were observed in HRV inoculated mini-pigs including small intestinal cell tumefaction and necrosis. HRV that was distributed in the small intestine was restricted to the top part of the villi on the internal wall of the ileum, which was observed by immunofluorescence and transmission electron microscopy. Virus particles were observed in Golgi like follicles in HRV-infected neonatal minipigs. Guizhou mini-pigs were more sensitive to HRV than bamma with respect to RV antigen shedding and clinical diarrhea.CONCLUSION: these results indicate that we have established a mini-pig model of HRV induced diarrhea. Our findings are useful for the understanding of the pathogenic mechanisms of HRV infection.
基金Supported by Chinese Center for Disease Control and Prevention and the State Key Laboratory for Infectious Disease Prevention and Control,No.2011SKLID103Beijing Key Laboratory of Environmental and Viral Oncology,College of Life Science and Bio-Engineering,Beijing University of Technology(Grant sponsor:State 863 projects,No.2012AA02A404,No2014ZX10005002 and No.PXM2014_014204_07_000046)
文摘AIM:to study the prevalence of human papillomavirus(HPV) in esophageal carcinoma in tangshan,China,a high-incidence area.METHODS:Formalin-fixed,paraffin-embedded tissue specimens from 198 patients who were pathologically diagnosed with esophageal squamous cell carcinoma from 2011 to 2013 were obtained from a pathology department in Tangshan.DNA was extracted from all198 specimens to detect HPV by polymerase chain reaction(PCR).β-globin PCR was performed to check the quality of the DNA extraction procedure.PCR was performed to detect a wide range of HPV types,and type-specific PCR was performed to detect HPV types16 and 18.Negative and positive controls were used for HPV 16 and 18 detection.RESULTS:the DNA extraction method in this study appeared to be more effective than other previously reported methods.After DNA extraction,more than98%of the tissue specimens had an acceptable result in the DNA qualification test(β-globin PCR).the overall prevalence of HPV in tumor tissues by GP6+/GP5+PCR was 79.79%,and the prevalence of HPV types16 and 18 was 40.40%and 47.47%,respectively.PCR demonstrated the presence of HPV,and direct sequencing confirmed the HPV genotypes.All HPVpositive PCR products were checked by DNA sequence analysis using DNAman and compared with the known HPV sequences listed in the basic Local Alignment Search tool database to evaluate the HPV types.this analysis confirmed the presence of HPV types 16 and18.CONCLUSION:DNA of high-risk HPV types 16 and 18is present in esophageal tumors,implicating HPV as a possible etiologic factor for esophageal squamous cell carcinoma.
文摘Correction to:Signal Transduction and Targeted Therapy https://doi.org/10.1038/s41392-022-01017-8,published online 22 June 2022 In the process of collating the raw data,the authors noticed one inadvertent mistake occurred in Fig.2e that needs to be corrected in the article1.The correct data are provided as follows.The key findings of the article are not affected by the correction.The original article has been corrected.
基金supported by grants from the National Natural Science Foundation of China(Grant No.82174202,No.32270407 and No.82074428)Natural Science Foundation of Sichuan Province(Grant No.2022NSFSC0726)+2 种基金Innovative Team Projects of Shanghai Municipal Commission of Health(Grant No.2022CX001)Innovation Foundation of the Affiliated Hospital of Chengdu University(Grant No.CDFYCX202209)Major Research and Development Plan of Xizang Autonomous Region(Grant No.XZ202201ZY0031G).
文摘Objective:To explore whether the ethanol extract of Herpetospermum caudigerum Wall(EHC),a Xizang medicinal plant traditionally used for treating liver diseases,can improve imiquimod-induced psoriasis-like skin inflammation.Methods:Immunohistochemistry and immunofluorescence staining were used to determine the effects of topical EHC use in vivo on the skin pathology of imiquimod-induced psoriasis in mice.The protein levels of interferon-γ(IFN-γ),tumor necrosis factor-a(TNF-a),and interleukin-17A(IL-17A)in mouse skin samples were examined using immunohistochemical staining.In vitro,IFN-γ-induced HaCaT cells with or without EHC treatment were used to evaluate the expression of keratinocyte-derived intercellular cell adhesion molecule-1(ICAM-1)and chemokine CXC ligand 9(CXCL9)using Western blotting and reverse transcription-quantitative polymerase chain reaction.The protein synthesis inhibitor cycloheximide and proteasome inhibitor MG132 were utilized to validate the EHC-mediated mechanism underlying degradation of ICAM-1 and CXCL9.Results:EHC improved inflammation in the imiquimod-induced psoriasis mouse model and reduced the levels of IFN-γ,TNF-a,and IL-17A in psoriatic lesions.Treatment with EHC also suppressed ICAM-1 and CXCL9 in epidermal keratinocytes.Further mechanistic studies revealed that EHC suppressed keratinocyte-derived ICAM-1 and CXCL9 by promoting ubiquitin–proteasome-mediated protein degradation rather than transcriptional repression.Seven primary compounds including ehletianol C,dehydrodiconiferyl alcohol,herpetrione,herpetin,herpetotriol,herpetetrone and herpetetrol were identified from the EHC using ultra-performance liquid chromatography-quadrupole-time of flight-mass spectrometry.Conclusion:Topical application of EHC ameliorates psoriasis-like skin symptoms and improves the inflammation at the lesion sites.
基金supported by grants from the National Natural Science Foundation of China,No.31100769
文摘Platelet-derived growth factor receptor alpha （PDGFRct） is a marker of oligodendrocyte precursor cells in the central nervous system. NG2 is also considered a marker of oligodendrocyte precursor cells. However, whether there are differences in the distribution and morphol- ogy of oligodendrocyte precursor cells labeled by NG2 or PDGFRa in the developing neonatal rat brain remains unclear. In this study, by immunohistochemical staining, NG2 positive （NG2＋） cells were ubiquitous in the molecular layer, external pyramidal layer, internal pyramidal layer, and polymorphic layer of the cerebral cortex, and corpus callosum, external capsule, piriform cortex, and medial septal nucleus. NG2~ cells were stellate or fusiform in shape with long processes that were progressively decreased and shortened over the course of brain development. The distribution and morphology of PDGFRct positive （PDGFRa＋） cells were coincident with NG2＋ cells. The co- localization of NG2 and PDGFRu in the cell bodies and processes of some cells was confirmed by double immunofluorescence labeling. Moreover, cells double-labeled for NG2 and PDGFRa were predominantly in the early postnatal stage of development. The numbers of NG2＋/PDGFRa＋ cells and PDGFRa＋ cells decreased, but the number of NG2＋ cells increased from postnatal days 3 to 14 in the developing brain. In addition, amoeboid microglial cells of the corpus callosum, newborn brain macrophages in the normal developing brain, did not express NG2 or PDGFRu, but NG2 expression was detected in amoeboid microglia after hypoxia. The present results suggest that NG2 and PDGFRct are specific markers of oligodendrocyte precursor cells at different stages during early development. Additionally, the NG2 protein is involved in inflammatory and pathological processes of amoeboid microglial cells.
基金supported by the Ministry of Science and Technology(2019YFA0801703)the National Natural Science Foundation of China(No.81790250,81790253,91959202 and 81802745)the Innovation Program of Shanghai Municipal Education Commission(N173606).
文摘Pancreatic ductal adenocarcinoma(PDAC)is well-known for inefficient early diagnosis,with most patients diagnosed at advanced stages.Increasing evidence indicates that elevated plasma levels of branched-chain amino acids(BCAAs)are associated with an increased risk of pancreatic cancer.Branched-chain amino acid transaminase 2(BCAT2)is an important enzyme in BCAA catabolism that reversibly catalyzes the initial step of BCAA degradation to branched-chain acyl-CoA.Here,we show that BCAT2 is acetylated at lysine 44(K44),an evolutionarily conserved residue.BCAT2 acetylation leads to its degradation through the ubiquitin–proteasome pathway and is stimulated in response to BCAA deprivation.cAMP-responsive element-binding(CREB)-binding protein(CBP)and SIRT4 are the acetyltransferase and deacetylase for BCAT2,respectively.CBP and SIRT4 bind to BCAT2 and control the K44 acetylation level in response to BCAA availability.More importantly,the K44R mutant promotes BCAA catabolism,cell proliferation,and pancreatic tumor growth.Collectively,the data from our study reveal a previously unknown regulatory mechanism of BCAT2 in PDAC and provide a potential therapeutic target for PDAC treatment.
基金the generous providing the plasmid of VCIP135-S1207A from Dr.Zhen-Kun Lou’s lab.27 We also appreciate the Biomedical Core Facility of Fudan University for technical supportsupported by National Key R&D Program of China(2020YFA0803402 and 2019YFA0801703 to Q.-Y.L.)+1 种基金National Natural Science Foundation of China(Nos.81790250/81790253,91959202 and 82121004 to Q.-Y.L.,No.81872240 to M.Y.,No.82002951 to J.-T.L.Innovation Program of Shanghai Municipal Education Commission(No.N173606 to Q.-Y.L.).
文摘Folic acid,served as dietary supplement,is closely linked to one-carbon metabolism and methionine metabolism.Previous clinical evidence indicated that folic acid supplementation displays dual effect on cancer development,promoting or suppressing tumor formation and progression.However,the underlying mechanism remains to be uncovered.Here,we report that high-folate diet significantly promotes cancer development in mice with hepatocellular carcinoma(HCC)induced by DEN/high-fat diet(HFD),simultaneously with increased expression of methionine adenosyltransferase 2A(gene name,MAT2A;protein name,MATIIα),the key enzyme in methionine metabolism,and acceleration of methionine cycle in cancer tissues.In contrast,folate-free diet reduces MATIIαexpression and impedes HFD-induced HCC development.Notably,methionine metabolism is dynamically reprogrammed with valosin-containing protein p97/p47 complex-interacting protein(VCIP135)which functions as a deubiquitylating enzyme to bind and stabilize MATIIαin response to folic acid signal.Consistently,upregulation of MATIIαexpression is positively correlated with increased VCIP135 protein level in human HCC tissues compared to adjacent tissues.Furthermore,liver-specific knockout of Mat2a remarkably abolishes the advocating effect of folic acid on HFD-induced HCC,demonstrating that the effect of high or free folate-diet on HFD-induced HCC relies on Mat2a.Moreover,folate and multiple intermediate metabolites in one-carbon metabolism are significantly decreased in vivo and in vitro upon Mat2a deletion.Together,folate promotes the integration of methionine and one-carbon metabolism,contributing to HCC development via hijacking MATIIαmetabolic pathway.This study provides insight into folate-promoted cancer development,strongly recommending the tailor-made folate supplement guideline for both sub-healthy populations and patients with cancer expressing high level of MATIIαexpression.
基金Project supported by the National Natural Science Foundation of China(Nos.51875518 and 51475419)the Key Research and Development Projects of Zhejiang Province(Nos.2017C01054 and2018C03062)the Fundamental Research Funds for the Central Universities(No.2019FZA4002),China
文摘Background:The shortage of donor corneas is a severe global issue,and hence the development of corneal alternatives is imperative and urgent.Although attempts to produce artificial cornea substitutes by tissue engineering have made some positive progress,many problems remain that hamper their clinical application worldwide.For example,the curvature of tissue-engineered cornea substitutes cannot be designed to fit the bulbus oculi of patients.Objective:To overcome these limitations,in this paper,we present a novel integrated three-dimensional(3 D) bioprintingbased cornea substitute fabrication strategy to realize design,customized fabrication,and evaluation of multi-layer hollow structures with complicated surfaces.Methods:The key rationale for this method is to combine digital light processing(DLP) and extrusion bioprinting into an integrated 3 D cornea bioprinting system.A designable and personalized corneal substitute was designed based on mathematical modelling and a computer tomography scan of a natural cornea.The printed corneal substitute was evaluated based on biomechanical analysis,weight,structural integrity,and fit.Results:The results revealed that the fabrication of high water content and highly transparent curved films with geometric features designed according to the natural human cornea can be achieved using a rapid,simple,and low-cost manufacturing process with a high repetition rate and quality.Conclusions:This study demonstrated the feasibility of customized design,analysis,and fabrication of a corneal substitute.The programmability of this method opens up the possibility of producing substitutes for other cornea-like shell structures with different scale and geometry features,such as the glomerulus,atrium,and oophoron.
基金supported by the National Key Research and Development Program of China (2020YFA0803402 and2019YFA0801703)the National Natural Science Foundation of China(81872240,81802745,81790250/81790253 and 91959202)Innovation Program of Shanghai Municipal Education Commission (N173606)。
文摘Epithelial ovarian cancer(EOC) exhibits strong dependency on the tricarboxylic acid(TCA) cycle and oxidative phosphorylation to fuel anabolic process.Here,we show that malate dehydrogenase 2(MDH2),a key enzyme of the TCA cycle,is palmitoylated at cysteine 138(C138) residue,resulting in increased activity of MDH2.We next identify that ZDHHC18 acts as a palmitoyltransferase of MDH2.Glutamine deprivation enhances MDH2 palmitoylation by increasing the binding between ZDHHC18 and MDH2.MDH2 silencing represses mitochondrial respiration as well as ovarian cancer cell proliferation both in vitro and in vivo.Intriguingly,re-expression of wild-type MDH2,but not its palmitoylation-deficient C138 S mutant,sustains mitochondrial respiration and restores the growth as well as clonogenic capability of ovarian cancer cells.Notably,MDH2 palmitoylation level is elevated in clinical cancer samples from patients with high-grade serous ovarian cancer.These observations suggest that MDH2 palmitoylation catalyzed by ZDHHC18 sustains mitochondrial respiration and promotes the malignancy of ovarian cancer,yielding possibilities of targeting ZDHHC18-mediated MDH2 palmitoylation in the treatment of EOC.
基金supported by the Industry-Academia-Research projects of Guangdong province, P. R. China (Grant Nos. 2014B090903012, 2013B090200008 and 2013B090600015)
文摘Electromagnetic field is an available online method to increase bonding strength of clad sheet manufactured by horizontal twin-roll casting （HTRC）. In this paper, an electric current pulse （ECP） and a complex field （static magnetic field （SMF） together with ECP） are exerted during HTRC of steel/aluminum clad sheet. The produced clad sheet has good appearance, and no visible defects exist at the bonding interface. The inter-diffusion zone at Fe/A1 interface in ECP and SMF＋ECP sheets is 3 and 4 μm, respectively, and the latter increases slightly compared with that in non-field sheet. The average peel strengths （APS） of ECP and SMF＋ECP sheet are 14 and 21 N/mm, respectively, which increase by 2 and 9 N/mm compared with 12 N/mm of non-field sheet. The APS increment in SMF＋ECP sheet is resulted from the increment of interface bonding spots and the enhancement of inter-diffusion zone width.
文摘To the Editor： Laparoendoscopic single-site surgery （LESS） has been widely used for various urologic diseases in order to improve safety, outcomes, and cosmesis, and to further reduce invasiveness. Here, we reported our initial experience with laparoendoscopic single-site retroperitoneal partial adrenalectomy using a homemade single-port device in a municipal hospital.
