Cadmium(Cd)contamination in soil poses a huge threat to plants even at low concentrations;Broussonetia papyrifera has great potential in remediation of soil heavy metal contamination.However,whether exogenous indole-3...Cadmium(Cd)contamination in soil poses a huge threat to plants even at low concentrations;Broussonetia papyrifera has great potential in remediation of soil heavy metal contamination.However,whether exogenous indole-3-acetic acid(IAA)application and arbuscular mycorrhizal fungi(AMF)have synergistic effects on Cd tolerance of B.papyrifera remains unclear.To investigate the effects of AMF inoculation and IAA application on the tolerance of B.papyrifera to Cd stress,two experiments were conducted:the first to investigate the effect of AMF(Rhizophagus irregularis)inoculation on the tolerance of B.papyrifera to Cd stress and the second to investigate the combined effects of AMF inoculation and IAA application on the tolerance of B.papyrifera to Cd stress.Parameters including endogenous hormone concentration,antioxidant defense response,malondialdehyde(MDA)content,and gene expression related to antioxidant enzyme system and hormone were measured.The results indicated that AMF alleviated Cd toxicity of B.papyrifera by reducing MDA content and improving antioxidant enzyme activities and Cd absorption capacity.Furthermore,the combination of AMF inoculation and IAA application had a synergetic effect on the tolerance of B.papyrifera to Cd stress through upregulating BpAUX1 and BpAUX2,which might contribute to root growth and root xylem synthesis,and by upregulating BpSOD2 and BpPOD34 to enhance the antioxidant enzyme system.This work provides a new insight into the application of IAA in the remediation of soil Cd pollution by mycorrhizal plants.展开更多
AIM： To investigate the possible mechanism for HBV X gene to induce apoptosis of hepatocyte HL-7702 cells.METHODS： HBV X gene eukaryon expression vector pcDNA3-X was established and transfected into HL-7702 cells by...AIM： To investigate the possible mechanism for HBV X gene to induce apoptosis of hepatocyte HL-7702 cells.METHODS： HBV X gene eukaryon expression vector pcDNA3-X was established and transfected into HL-7702 cells by lipid-mediated transfection, including transient and stable transfection. Positive clones were screened by incubating in the selective medium with 600 μg/mL G418 and named HL-7702/HBV-encoded X protein （HBx） cells. The expressions of Fas/FasL, Bax/Bcl-2, and c-myc mRNA were measured by semi-quantitative RT-PCR in HL-7702/HBx and control group, respectively.RESULTS： RT-PCR analysis confirmed that HBV X gene was transfected into HL-7702 cells successfully. By semiquantitative RT-PCR analysis, Bax and c-myc mRNA levels in HL-7702/HBx cells of transient transfection were significantly higher than those in control, FasL and c-myc mRNA levels in HL-7702/HBx cells of stable transfection were significantly higher than those in control, whereas the Bcl-2 mRNA levels in HL-7702/HBx cells of transient and stable transfection were significantly lower than thosein control.CONCLUSION： HBV X gene may promote the apoptosis of hepatocytes by regulating the expressions of Fas/FasL, Bax/Bcl-2, and c-myc gene in a dose-dependent manner.展开更多
Heat and acid treatments were reported to be a promising substitute for SO2 fumigation in color protection of postharvest lychee （Litchi chinensis） fruits, but the mechanism was not clear. In the present study, hot ...Heat and acid treatments were reported to be a promising substitute for SO2 fumigation in color protection of postharvest lychee （Litchi chinensis） fruits, but the mechanism was not clear. In the present study, hot water （70℃） dipping followed by immersion in 2% HC1 （heat-acid） substantially protected the red color of the fruit during storage at 25℃ and inhibited anthocyanin degradation while hot water dipping alone （heat） led to rapidly browning and about 90% loss in anthocyanin content. The pH values in the pericarp of the heat-acid treated fruit dropped to 3.2, while the values maintained around 5.0 in the heat-treated and control fruit. No significantly different pH values were detected among the arils of heat-acid, heat treated and control fruit. Heat-acid treatment dramatically reduced the activities of anthocyanin degradation enzyme （ADE）, peroxidase （POD） and polyphenol oxidase in the pericarp. A marked reduction in LcPOD gene expression was also detected in heat-acid treated fruit, in contrast, induction was found in heat treated fruit. The pericarp of heat-acid treated fruit exhibited significantly lower respiration rate but faster water loss than that of the untreated or heat treated fruit. Taken together, heat treatment triggered quick browning and anthocyanin loss in lychee fruit, while heat-acid treatment protected the fruit color by a great reduction in the activities/gene expression of anthocyanin degradation enzymes and acidification of lychee pericarp.展开更多
AIM:To evaluate for the first time the protein and mRNA expression of 14-3-3εin gastric carcinogenesis.METHODS:14-3-3εprotein expression was determined by western blotting,and mRNA expression was examined by real-ti...AIM:To evaluate for the first time the protein and mRNA expression of 14-3-3εin gastric carcinogenesis.METHODS:14-3-3εprotein expression was determined by western blotting,and mRNA expression was examined by real-time quantitative RT-PCR in gastric tumors and their matched non-neoplastic gastric tissue samples.RESULTS:Authors observed a significant reduction of 14-3-3εprotein expression in gastric cancer(GC)samples compared to their matched non-neoplastic tissue.Reduced levels of 14-3-3εwere also associated with diffuse-type GC and early-onset of this pathology.Our data suggest that reduced 14-3-3εmay have a role in gastric carcinogenesis process.CONCLUSION:Our results reveal that the reduced 14-3-3εexpression in GC and investigation of 14-3-3ε interaction partners may help to elucidate the carcino-genesis process.展开更多
Spatial transcriptomics enables the study of localization-indexed gene expression activity in tissues,providing the transcriptional landscape that in turn indicates the potential regulatory networks of gene expression...Spatial transcriptomics enables the study of localization-indexed gene expression activity in tissues,providing the transcriptional landscape that in turn indicates the potential regulatory networks of gene expression.In situ sequencing(ISS)is a targeted spatial transcriptomic technique,based on padlock probe and rolling circle amplification combined with next-generation sequencing chemistry,for highly multiplexed in situ gene expression profiling.Here,we present improved in situ sequencing(IISS)that exploits a new probing and barcoding approach,combined with advanced image analysis pipelines for high-resolution targeted spatial gene expression profiling.We develop an improved combinatorial probe anchor ligation chemistry using a 2-base encoding strategy for barcode interrogation.The new encoding strategy results in higher signal intensity as well as improved specificity for in situ sequencing,while maintaining a streamlined analysis pipeline for targeted spatial transcriptomics.We show that IISS can be applied to both fresh frozen tissue and formalin-fixed paraffin-embedded tissue sections for single-cell level spatial gene expression analysis,based on which the developmental trajectory and cell-cell communication networks can also be constructed.展开更多
This paper deals with the estimation of crest settlement in a concrete face rockfill dam （CFRD）, utilizing intelligent methods. Following completion of dam construction, considerable movements of the crest and the b...This paper deals with the estimation of crest settlement in a concrete face rockfill dam （CFRD）, utilizing intelligent methods. Following completion of dam construction, considerable movements of the crest and the body of the dam can develop during the first impoundment of the reservoir. Although there is vast experience worldwide in CFRD design and construction, few accurate experimental relationships are available to predict the settlement in CFRD. The goal is to advance the development of intelligent methods to estimate the subsidence of dams at the design stage. Due to dam zonifieation and uncertainties in material properties, these methods appear to be the appropriate choice. In this study, the crest settlement behavior of CFRDs is analyzed based on compiled data of 24 CFRDs constructed during recent years around the world, along with the utilization of gene ex- pression programming （GEP） and adaptive neuro-fuzzy inference system （ANFIS） methods. In addition, dam height （H）, shape factor （St）, and time （t, time after first operation） are also assessed, being considered major factors in predicting the settlement behavior. From the relationships proposed, the values ofR2 for both equations of GEP （with and without constant） were 0.9603 and 0.9734, and for the three approaches of ANFIS （grid partitioning （GP）, subtractive clustering method （SCM）, and fuzzy c-means clustering （FCM）） were 0.9693, 0.8657, and 0.8848, respectively. The obtained results indicate that the overall behavior evaluated by this approach is consistent with the measured data of other CFRDs.展开更多
The neuromuscular junction becomes progressively less receptive to regenerating axons if nerve repair is delayed for a long period of time. It is difficult to ascertain the denervated muscle＇s residual receptivity by...The neuromuscular junction becomes progressively less receptive to regenerating axons if nerve repair is delayed for a long period of time. It is difficult to ascertain the denervated muscle＇s residual receptivity by time alone. Other sensitive markers that closely correlate with the extent of denervation should be found. After a denervated muscle develops a fibrillation potential, muscle fiber conduction velocity, muscle fiber diameter, muscle wet weight, and maximal isometric force all decrease; remodeling increases neuromuscular junction fragmentation and plantar area, and expression of myogenesis-related genes is initially up-regulated and then down-regulated. All these changes correlate with both the time course and degree of denervation. The nature and time course of these denervation changes in muscle are reviewed from the literature to explore their roles in assessing both the degree of detrimental changes and the potential success of a nerve repair. Fibrillation potential amplitude, muscle fiber conduction velocity, muscle fiber diameter, mRNA expression levels of myogenic regulatory factors and nicotinic acetylcholine receptor could all reflect the severity and length of denervation and the receptiveness of denervated muscle to regenerating axons, which could possibly offer an important clue for surgical choices and predict the outcomes of delayed nerve repair.展开更多
AIM： A few studies have applied genomic-wide gene expression analysis in inflamed colon tissue sample in ulcerative colitis （UC）. We reported the first study of non-inflamed mucosal gene expression in UC and explor...AIM： A few studies have applied genomic-wide gene expression analysis in inflamed colon tissue sample in ulcerative colitis （UC）. We reported the first study of non-inflamed mucosal gene expression in UC and explored its clinical implication. METHODS： Non-inflamed mucosa was obtained from 6 UC patients who received total colectomy. The gene expression of UC in noninflamed mucosa was monitored with a microarray. For a selected gene, RT-PCR was performed to verify array results and to further examine expression pattern in inflamed mucosa of UC, colorectal cancer tissue and normal mucosa using additional matched pairs. RESULTS： Two genes showing 2.5-fold decreased expression with significance set at in UC samples were borneo box a4 （HOXa4） and mads box transcription enhancer factor 2, polypeptide B （MEF2b）. RT-PCR showed that MEF2b expression in non-inflamed mucosa was significantly downregulated, whereas the expression of MEF2b increased in accordance with the severity of mucosal inflammation. HOXa4 expression both in inflamed and non-inflamed mucosa in UC was consistently downregulated, and the downregulation of HOXa4 was also found in colorectal carcinoma. CONCLUSION： It is suggested that the MEF2b expression in UC which increase in accordance with inflammation may be induced by the inflammatory mediator. In contrast the downregulation of HOXa4 may be partly involved in the pathogenesis of disease including UC and UC-related carcinogenesis.展开更多
To investigate the molecular mechanism underlying the neuroprotective effect of lithium on cells, in this study, we exposed SH-SY5Y cells to 0.5 mmol/L lithium carbonate （Li2CO2） for 25-50 weeks and then detected th...To investigate the molecular mechanism underlying the neuroprotective effect of lithium on cells, in this study, we exposed SH-SY5Y cells to 0.5 mmol/L lithium carbonate （Li2CO2） for 25-50 weeks and then detected the expression levels of some neurobiology related genes and post-translational modifications of stress proteins in SH-SYSY cells, cDNA arrays showed that pyruvate kinase 2 （PKM2） and calmodulin 3 （CaM 3） expression levels were significantly down-regulated, phosphatase protein PP2A expression was lightly down-regulated, and casein kinase II （CK2）, threonine/tyrosine phosphatase 7 （PYST2）, and dopamine beta-hydroxylase （DBH） expression levels were significantly up-regulated. Besides, western blot analysis of stress proteins （HSP27, HSP70, GRP78 and GRP94） showed an over-expression of two proteins： a 105 kDa protein which is a hyper-phosphorylated isoform of GRP94, and a 108 kDa protein which is a phosphorylated tetramer of HSP27. These results suggest that the neuroprotective effects of lithium are likely related to gene expressions and post-translational modifications of proteins cited above.展开更多
AIM： To elucidate the molecular mechanisms of the inhibitory effects of IFN-α on tumor growth and metastasis in MHCC97 xenografts. METHODS： Three thousand milliliter of IFN-α-treated and international units per -u...AIM： To elucidate the molecular mechanisms of the inhibitory effects of IFN-α on tumor growth and metastasis in MHCC97 xenografts. METHODS： Three thousand milliliter of IFN-α-treated and international units per -untreated MHCC97 cells were enrolled for gene expression analysis using cDNA microarray. The mRNA levels of several differentially expressed genes in cDNA microarray were further identified by Northern blot and RT-PCR. RESULTS- A total of 190 differentially expressed genes including 151 IFN-α-repressed and 39 -stimulated genes or expressed sequence tags from 8 464 known human genes were found to be regulated by IFN-α in MHCC97. With a few exceptions, mRNA levels of the selected genes in RT-PCR and Northern blot were in good agreement with those in cDNA microarray. CONCLUSION： IFN-α might exert its complicated anti- tumor effects on MHCC97 xenografts by regulating the expression of functional genes involved in cell metabolism, proliferation, morphogenesis, angiogenesis, and signaling.展开更多
The Wnt/β-catenin signaling pathway plays a crucial role in the embryonic development of metazoans. Although the pathway has been studied extensively in many model animals, its function in amphioxus, the most primiti...The Wnt/β-catenin signaling pathway plays a crucial role in the embryonic development of metazoans. Although the pathway has been studied extensively in many model animals, its function in amphioxus, the most primitive chordate, remains largely uncharacterized. To obtain basic data for functional analysis, we identified and isolated seven genes （Lrp5/6, Dvl, APC, Ckla, CklS, Gsk3β, and Gro） of the Wnt/β-catenin signaling pathway from the amphioxus （Branchiostoma floridae） genome. Phylogenetic analysis revealed that amphioxus had fewer members of each gene family than that found in vertebrates. Whole-mount in situ hybridization showed that the genes were maternally expressed and broadly distributed throughout the whole embryo at the cleavage and blastula stages. Among them, Dvl was expressed asymmetrically towards the animal pole, while the others were evenly distributed in all blastomeres. At the mid-gastrula stage, the genes were specifically expressed in the primitive endomesoderm, but displayed different patterns. When the embryo developed into the neurula stage, the gene expressions were mainly detected in either paraxial somites or the tail bud. With the development of the embryo, the expression levels further decreased gradually and remained only in some pharyngeal regions or the tail bud at the larva stage. Our results suggest that the Wnt/β-catenin pathway might be involved in amphioxus somite formation and posterior growth, but not in endomesoderm specification.展开更多
Objective:To investigate the involvement of reactive oxygen species detoxification system in Anopheles stephensi during Plasmodium berghei midgut invasion.Methods:Eight key reactive oxygen species metabolizing enzymes...Objective:To investigate the involvement of reactive oxygen species detoxification system in Anopheles stephensi during Plasmodium berghei midgut invasion.Methods:Eight key reactive oxygen species metabolizing enzymes were cloned and characterized,and their expression was monitored in parasite-infected mosquitoes.Results:Superoxide anion detoxifying superoxide dismutases(Fe/Mn SOD,Cu/Zn SOD 2,Cu/Zn SOD 3A,and Cu/Zn SOD 3B)depicted varied expression patterns.Fe/Mn SOD expression declined,whereas Cu/Zn SOD expression was elevated in the infected mosquitoes.Peroxidases,catalase and glutathione peroxidase showed lack of induction in expression during the Plasmodium berghei infection.Further,expression of thioredoxin reductase increased in the infected mosquitoes,whereas gluthathione S-transferase levels decreased markedly.Conclusions:Detoxification enzymes may play a role in modulating host immunity and parasite transmission.展开更多
Objective: To study the molecular mechanism ofdifferent tumorigenicity in nude mice of human leukemiacell lines K562-n and K562. Methods: To analyze the genes differently expressed between K562 and K562-n cells byusin...Objective: To study the molecular mechanism ofdifferent tumorigenicity in nude mice of human leukemiacell lines K562-n and K562. Methods: To analyze the genes differently expressed between K562 and K562-n cells byusing cDNA microarray technique. Results: Among the12800 genes detected, some genes involved in materialmetabolism and material transport were differentlyexpressed between K562-n and K562 cells. These genesinclude homo sapiens placenta-specific ATP-binding cassette transporter gene, dihydrodiol dehydrogenase gene, hepatic dihydrodiol dehydrogenase gene, NAD-dependent methylene tetrahydrofolate dehydrogenase cyclohydrolase,lysophosphatidic acid acyltransferase, alpha gene,argininosuccinate lyase gene, mitochondrial isocitrtatedehydrogenase, adhesion protein SQM1 gene, dimethylarginine dimethylamino-hydrolase gene, M1subunit of ribonucleotide reductase and farnesylpyrophosphate synthetase gene. Conclusion: The hightumorigenicity of K562-n cells is related to the differentexpression of some genes concerned with cell metabolismand material transpoert.展开更多
The full-length cDNA encoding the subunits p40 and p35 of human interleukin12(hIL12) were cloned separately by RTPCR, linked together by internal ribosomal entry site (IRES) of encephalomyocarditis virus which initiat...The full-length cDNA encoding the subunits p40 and p35 of human interleukin12(hIL12) were cloned separately by RTPCR, linked together by internal ribosomal entry site (IRES) of encephalomyocarditis virus which initiates capindependent translation to form a dicistronic gene fragment. The dicistronic fragment was placed between the cytomegalovirus (CMV) promoter and SV40 polyA signal to form a dicistronic expression cassette. Subsequently, the dicistronic expression cassette was inserted into E1 region of Ad5 genome in cosmid vector pAx1cw of E1substitution type. By homologous recombination with EcoT22Idigested Ad5 DNATPC in 293 cells, the replicationdeficient recombinant adenoviruses of hIL12 were generated efficiently. After infected with hIL12 recombinant adenoviruses in vitro, 293 cells, human hepatocellular carcinoma cells HepG2, and primary human skin fibroblasts expressed and secreted hIL12 at comparable levels (30～60ng/ 106cells/24hr), which could stimulate the proliferation and IFNγ production of human lymphoblasts. These suggest that the dicistronic adenovirus vector of hIL12 could effectively mediate the expression of bioactive hIL12 and might be used in cancer gene therapy.展开更多
Chitin is a common component in the natural diet of many fish.While a range of chitin-degrading chitinases have been identified,it is unclear whether chitin is metabolized in fish.Here we use a combination of chitinas...Chitin is a common component in the natural diet of many fish.While a range of chitin-degrading chitinases have been identified,it is unclear whether chitin is metabolized in fish.Here we use a combination of chitinase activity assays,transcriptomics,and 16S rRNA bacterial analysis to assess the effect of chitin supplementation on Atlantic salmon gene expression and microbial community composition.Atlantic salmon express multiple genes associated with chitin metabolism.We show that the expression and activity of Atlantic salmon chitinases are not affected by the addition of dietary chitin,but we demonstrate an association between gut microbial composition,chitinase activity in the gut,and host chitinase expression.These results contribute to a greater understanding of chitin metabolism in fish.展开更多
Objective： To evaluate the effects and possible mechanisms of rutaecarpine on angiotensin Ⅱ （Ang Ⅱ ）-induced proliferation in cultured rat vascular smooth muscle cells （VSMCs）. Methods： VSMCs were isolated fro...Objective： To evaluate the effects and possible mechanisms of rutaecarpine on angiotensin Ⅱ （Ang Ⅱ ）-induced proliferation in cultured rat vascular smooth muscle cells （VSMCs）. Methods： VSMCs were isolated from Male Sprague-Dawley rat aorta, and cultured by enzymic dispersion method. Experiments were performed with cells from passages 3-8. The cultured VSMCs were randomly divided into control, model （Ang Ⅱ 0.1 μ moVL）, and rutaecarpine （0.3-3.0μmol/L） groups. VMSC proliferation was induced by Ang Ⅱ, and was evaluated by the 3-（4, 5-dimethylthiazol-2-yl）-2, 5-diphenyltetrazolium bromide assay and cell counting. To examine the mechanisms involved in anti-proliferative effects of rutaecarpine, nitric oxide （NO） levels and NO synthetase （NOS） activity were determined. Expressions of VSMC proliferation-related genes including endothelial nitric oxide synthase （eNOS）, and c-myc hypertension related gene-1 （HRG-1） were determined by real-time reverse chain reaction （RT-PCR）. Results： Rutaecarpine （0.3-3.0μmol/l_） inhibited Ang R-induced VSMC proliferation and the best effects were achieved at 3.0 μmol/L. The Ang Ⅱ-induced decreases in cellular NO contents and NOS activities were antagonized by rutaecarpine （P〈0.05）. Ang Ⅱ administration suppressed the expressions of eNOS and HRG-1, while increased c-myc expression （P〈0.05）. All these effects were attenuated by 3.0μmol/L rutaecarpine （P〈0.05）. Conclusion： Rutaecarpine is effective against Ang Ⅱ-induced rat VSMC proliferation, and this effect is due, at least in part, to NO production and the modulation of VMSC proliferation-related gene expressions.展开更多
Class B scavenger receptors （SR-Bs） are cell surface glycoproteins involved in various physiological processes in vivo, including the transport and metabolism of lipids, binding and phagoeytosis of xenobiotics, and ...Class B scavenger receptors （SR-Bs） are cell surface glycoproteins involved in various physiological processes in vivo, including the transport and metabolism of lipids, binding and phagoeytosis of xenobiotics, and signaling. But little information is available about silkworm SR-Bs; it is necessary to study these SR-Bs for revealing their function. In this study, we cloned the full-length coding sequence of BrnSCRBQ4, a SR-B gene from the silkworm Bombyx mori L. We found that the BmSCRBQ4 gene consists of nine exons and eight introns, with an open reading frame of 1371 bp encoding 456 amino acids. Gene expression studies determined that BmSCRBQ4 messenger RNA （mRNA） was expressed in unfertilized eggs, during embryonic development and throughout the majority of the larval period. Expression of mRNA was detected in the mid gut, middle silk gland, posterior silk gland, head, integumentum, fat body, testes and the ovaries of the larval B. mori Dazao strain, as well as in the silkworm cell lines BmN and BmE. Protein expression studies found BmSCRBQ4 protein was expressed only in the testes, fat body and middle silk gland of larvae, as well as in the silkworm cell lines BmN and BmE. The BmSCRBQ4 protein showed variability in banding patterns in different tissues and cells when analyzed by Western blotting. Immunohistochemical staining showed that the BmSCRBQ4 protein localizes to the constitutive membranes or cellular membranes of these tissues. These results indicated that BmSCRBQ4 gene may play some physiologically relevant roles at the cell surface in each tissue.展开更多
Immunotherapy is a promising cancer treatment method;however,only a few patients benefit from it.The development of new immunotherapy strategies and effective biomarkers of response and resistance is urgently needed.R...Immunotherapy is a promising cancer treatment method;however,only a few patients benefit from it.The development of new immunotherapy strategies and effective biomarkers of response and resistance is urgently needed.Recently,high-throughput bulk and single-cell gene expression profling technologies have generated valuable resources.However,these resources are not well organized and systematic analysis is difficult.Here,we present TIGER,a tumor immunotherapy gene expression resource,which contains bulk transcriptome data of 1508 tumor samples with clinical immunotherapy outcomes and 11,057 tumor/normal samples without clinical immunotherapy outcomes,as well as single-cell transcriptome data of 2,116,945 immune cells from 655 samples.TIGER provides many useful modules for analyzing collected and user-provided data.Using the resource in TIGER,we identified a tumor-enriched subset of CD4^(+)T cells.Patients with melanoma with a higher signature score of this subset have a significantly better response and survival under immunotherapy.We believe that TIGER will be helpful in understanding anti-tumor immunity mechanisms and discovering effective biomarkers.展开更多
基金supported by the National Natural Science Foundation of China(Nos.32001289 and 32071639)the Laboratory of Lingnan Modern Agriculture Project,China(No.NZ2021025)。
文摘Cadmium(Cd)contamination in soil poses a huge threat to plants even at low concentrations;Broussonetia papyrifera has great potential in remediation of soil heavy metal contamination.However,whether exogenous indole-3-acetic acid(IAA)application and arbuscular mycorrhizal fungi(AMF)have synergistic effects on Cd tolerance of B.papyrifera remains unclear.To investigate the effects of AMF inoculation and IAA application on the tolerance of B.papyrifera to Cd stress,two experiments were conducted:the first to investigate the effect of AMF(Rhizophagus irregularis)inoculation on the tolerance of B.papyrifera to Cd stress and the second to investigate the combined effects of AMF inoculation and IAA application on the tolerance of B.papyrifera to Cd stress.Parameters including endogenous hormone concentration,antioxidant defense response,malondialdehyde(MDA)content,and gene expression related to antioxidant enzyme system and hormone were measured.The results indicated that AMF alleviated Cd toxicity of B.papyrifera by reducing MDA content and improving antioxidant enzyme activities and Cd absorption capacity.Furthermore,the combination of AMF inoculation and IAA application had a synergetic effect on the tolerance of B.papyrifera to Cd stress through upregulating BpAUX1 and BpAUX2,which might contribute to root growth and root xylem synthesis,and by upregulating BpSOD2 and BpPOD34 to enhance the antioxidant enzyme system.This work provides a new insight into the application of IAA in the remediation of soil Cd pollution by mycorrhizal plants.
基金Supported by the Science and Technology Fund of Fujian Province, No. 99-Z-162
文摘AIM： To investigate the possible mechanism for HBV X gene to induce apoptosis of hepatocyte HL-7702 cells.METHODS： HBV X gene eukaryon expression vector pcDNA3-X was established and transfected into HL-7702 cells by lipid-mediated transfection, including transient and stable transfection. Positive clones were screened by incubating in the selective medium with 600 μg/mL G418 and named HL-7702/HBV-encoded X protein （HBx） cells. The expressions of Fas/FasL, Bax/Bcl-2, and c-myc mRNA were measured by semi-quantitative RT-PCR in HL-7702/HBx and control group, respectively.RESULTS： RT-PCR analysis confirmed that HBV X gene was transfected into HL-7702 cells successfully. By semiquantitative RT-PCR analysis, Bax and c-myc mRNA levels in HL-7702/HBx cells of transient transfection were significantly higher than those in control, FasL and c-myc mRNA levels in HL-7702/HBx cells of stable transfection were significantly higher than those in control, whereas the Bcl-2 mRNA levels in HL-7702/HBx cells of transient and stable transfection were significantly lower than thosein control.CONCLUSION： HBV X gene may promote the apoptosis of hepatocytes by regulating the expressions of Fas/FasL, Bax/Bcl-2, and c-myc gene in a dose-dependent manner.
基金supported by the National Key Basic Research Program of China (2013CB127105)the National Natural Science Foundation of China (30671466)+1 种基金China Litchi and Logan Research System (CARS-33-14)Guangdong Fruit Research System,China (2009-356)
文摘Heat and acid treatments were reported to be a promising substitute for SO2 fumigation in color protection of postharvest lychee （Litchi chinensis） fruits, but the mechanism was not clear. In the present study, hot water （70℃） dipping followed by immersion in 2% HC1 （heat-acid） substantially protected the red color of the fruit during storage at 25℃ and inhibited anthocyanin degradation while hot water dipping alone （heat） led to rapidly browning and about 90% loss in anthocyanin content. The pH values in the pericarp of the heat-acid treated fruit dropped to 3.2, while the values maintained around 5.0 in the heat-treated and control fruit. No significantly different pH values were detected among the arils of heat-acid, heat treated and control fruit. Heat-acid treatment dramatically reduced the activities of anthocyanin degradation enzyme （ADE）, peroxidase （POD） and polyphenol oxidase in the pericarp. A marked reduction in LcPOD gene expression was also detected in heat-acid treated fruit, in contrast, induction was found in heat treated fruit. The pericarp of heat-acid treated fruit exhibited significantly lower respiration rate but faster water loss than that of the untreated or heat treated fruit. Taken together, heat treatment triggered quick browning and anthocyanin loss in lychee fruit, while heat-acid treatment protected the fruit color by a great reduction in the activities/gene expression of anthocyanin degradation enzymes and acidification of lychee pericarp.
基金Supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico(CNPqChammas R,Smith MAC and Burbano RR)+1 种基金Fundao de Amparoà Pesquisa do Estado de So Paulo (FAPESPLeal MF and Calcagno DQ)
文摘AIM:To evaluate for the first time the protein and mRNA expression of 14-3-3εin gastric carcinogenesis.METHODS:14-3-3εprotein expression was determined by western blotting,and mRNA expression was examined by real-time quantitative RT-PCR in gastric tumors and their matched non-neoplastic gastric tissue samples.RESULTS:Authors observed a significant reduction of 14-3-3εprotein expression in gastric cancer(GC)samples compared to their matched non-neoplastic tissue.Reduced levels of 14-3-3εwere also associated with diffuse-type GC and early-onset of this pathology.Our data suggest that reduced 14-3-3εmay have a role in gastric carcinogenesis process.CONCLUSION:Our results reveal that the reduced 14-3-3εexpression in GC and investigation of 14-3-3ε interaction partners may help to elucidate the carcino-genesis process.
基金supported by the Natural Science Foundation of Fujian Province(2022J06022)the Quanzhou Science and Technology Plan Project(2021C040R)the Scientific Research Funds of Huaqiao University.
文摘Spatial transcriptomics enables the study of localization-indexed gene expression activity in tissues,providing the transcriptional landscape that in turn indicates the potential regulatory networks of gene expression.In situ sequencing(ISS)is a targeted spatial transcriptomic technique,based on padlock probe and rolling circle amplification combined with next-generation sequencing chemistry,for highly multiplexed in situ gene expression profiling.Here,we present improved in situ sequencing(IISS)that exploits a new probing and barcoding approach,combined with advanced image analysis pipelines for high-resolution targeted spatial gene expression profiling.We develop an improved combinatorial probe anchor ligation chemistry using a 2-base encoding strategy for barcode interrogation.The new encoding strategy results in higher signal intensity as well as improved specificity for in situ sequencing,while maintaining a streamlined analysis pipeline for targeted spatial transcriptomics.We show that IISS can be applied to both fresh frozen tissue and formalin-fixed paraffin-embedded tissue sections for single-cell level spatial gene expression analysis,based on which the developmental trajectory and cell-cell communication networks can also be constructed.
文摘This paper deals with the estimation of crest settlement in a concrete face rockfill dam （CFRD）, utilizing intelligent methods. Following completion of dam construction, considerable movements of the crest and the body of the dam can develop during the first impoundment of the reservoir. Although there is vast experience worldwide in CFRD design and construction, few accurate experimental relationships are available to predict the settlement in CFRD. The goal is to advance the development of intelligent methods to estimate the subsidence of dams at the design stage. Due to dam zonifieation and uncertainties in material properties, these methods appear to be the appropriate choice. In this study, the crest settlement behavior of CFRDs is analyzed based on compiled data of 24 CFRDs constructed during recent years around the world, along with the utilization of gene ex- pression programming （GEP） and adaptive neuro-fuzzy inference system （ANFIS） methods. In addition, dam height （H）, shape factor （St）, and time （t, time after first operation） are also assessed, being considered major factors in predicting the settlement behavior. From the relationships proposed, the values ofR2 for both equations of GEP （with and without constant） were 0.9603 and 0.9734, and for the three approaches of ANFIS （grid partitioning （GP）, subtractive clustering method （SCM）, and fuzzy c-means clustering （FCM）） were 0.9693, 0.8657, and 0.8848, respectively. The obtained results indicate that the overall behavior evaluated by this approach is consistent with the measured data of other CFRDs.
基金sponsored by the Armed Forces Institute of Regenerative Medicine award number W81XWH-08-2-0034supported by the Sundt Fellowship fund,Department of Neurologic Surgery,Mayo Clinic,USA
文摘The neuromuscular junction becomes progressively less receptive to regenerating axons if nerve repair is delayed for a long period of time. It is difficult to ascertain the denervated muscle＇s residual receptivity by time alone. Other sensitive markers that closely correlate with the extent of denervation should be found. After a denervated muscle develops a fibrillation potential, muscle fiber conduction velocity, muscle fiber diameter, muscle wet weight, and maximal isometric force all decrease; remodeling increases neuromuscular junction fragmentation and plantar area, and expression of myogenesis-related genes is initially up-regulated and then down-regulated. All these changes correlate with both the time course and degree of denervation. The nature and time course of these denervation changes in muscle are reviewed from the literature to explore their roles in assessing both the degree of detrimental changes and the potential success of a nerve repair. Fibrillation potential amplitude, muscle fiber conduction velocity, muscle fiber diameter, mRNA expression levels of myogenic regulatory factors and nicotinic acetylcholine receptor could all reflect the severity and length of denervation and the receptiveness of denervated muscle to regenerating axons, which could possibly offer an important clue for surgical choices and predict the outcomes of delayed nerve repair.
文摘AIM： A few studies have applied genomic-wide gene expression analysis in inflamed colon tissue sample in ulcerative colitis （UC）. We reported the first study of non-inflamed mucosal gene expression in UC and explored its clinical implication. METHODS： Non-inflamed mucosa was obtained from 6 UC patients who received total colectomy. The gene expression of UC in noninflamed mucosa was monitored with a microarray. For a selected gene, RT-PCR was performed to verify array results and to further examine expression pattern in inflamed mucosa of UC, colorectal cancer tissue and normal mucosa using additional matched pairs. RESULTS： Two genes showing 2.5-fold decreased expression with significance set at in UC samples were borneo box a4 （HOXa4） and mads box transcription enhancer factor 2, polypeptide B （MEF2b）. RT-PCR showed that MEF2b expression in non-inflamed mucosa was significantly downregulated, whereas the expression of MEF2b increased in accordance with the severity of mucosal inflammation. HOXa4 expression both in inflamed and non-inflamed mucosa in UC was consistently downregulated, and the downregulation of HOXa4 was also found in colorectal carcinoma. CONCLUSION： It is suggested that the MEF2b expression in UC which increase in accordance with inflammation may be induced by the inflammatory mediator. In contrast the downregulation of HOXa4 may be partly involved in the pathogenesis of disease including UC and UC-related carcinogenesis.
文摘To investigate the molecular mechanism underlying the neuroprotective effect of lithium on cells, in this study, we exposed SH-SY5Y cells to 0.5 mmol/L lithium carbonate （Li2CO2） for 25-50 weeks and then detected the expression levels of some neurobiology related genes and post-translational modifications of stress proteins in SH-SYSY cells, cDNA arrays showed that pyruvate kinase 2 （PKM2） and calmodulin 3 （CaM 3） expression levels were significantly down-regulated, phosphatase protein PP2A expression was lightly down-regulated, and casein kinase II （CK2）, threonine/tyrosine phosphatase 7 （PYST2）, and dopamine beta-hydroxylase （DBH） expression levels were significantly up-regulated. Besides, western blot analysis of stress proteins （HSP27, HSP70, GRP78 and GRP94） showed an over-expression of two proteins： a 105 kDa protein which is a hyper-phosphorylated isoform of GRP94, and a 108 kDa protein which is a phosphorylated tetramer of HSP27. These results suggest that the neuroprotective effects of lithium are likely related to gene expressions and post-translational modifications of proteins cited above.
基金Supported by the Key Projects for the Clinical Medicine from the Ministry of Public Health of China (2002-2005)
文摘AIM： To elucidate the molecular mechanisms of the inhibitory effects of IFN-α on tumor growth and metastasis in MHCC97 xenografts. METHODS： Three thousand milliliter of IFN-α-treated and international units per -untreated MHCC97 cells were enrolled for gene expression analysis using cDNA microarray. The mRNA levels of several differentially expressed genes in cDNA microarray were further identified by Northern blot and RT-PCR. RESULTS- A total of 190 differentially expressed genes including 151 IFN-α-repressed and 39 -stimulated genes or expressed sequence tags from 8 464 known human genes were found to be regulated by IFN-α in MHCC97. With a few exceptions, mRNA levels of the selected genes in RT-PCR and Northern blot were in good agreement with those in cDNA microarray. CONCLUSION： IFN-α might exert its complicated anti- tumor effects on MHCC97 xenografts by regulating the expression of functional genes involved in cell metabolism, proliferation, morphogenesis, angiogenesis, and signaling.
基金financially supported by the National Natural Science Foundation of China(31372188,31471986)the Science and Technology Innovation Commission of Shenzhen Municipality(CXZZ20120614164555920)
文摘The Wnt/β-catenin signaling pathway plays a crucial role in the embryonic development of metazoans. Although the pathway has been studied extensively in many model animals, its function in amphioxus, the most primitive chordate, remains largely uncharacterized. To obtain basic data for functional analysis, we identified and isolated seven genes （Lrp5/6, Dvl, APC, Ckla, CklS, Gsk3β, and Gro） of the Wnt/β-catenin signaling pathway from the amphioxus （Branchiostoma floridae） genome. Phylogenetic analysis revealed that amphioxus had fewer members of each gene family than that found in vertebrates. Whole-mount in situ hybridization showed that the genes were maternally expressed and broadly distributed throughout the whole embryo at the cleavage and blastula stages. Among them, Dvl was expressed asymmetrically towards the animal pole, while the others were evenly distributed in all blastomeres. At the mid-gastrula stage, the genes were specifically expressed in the primitive endomesoderm, but displayed different patterns. When the embryo developed into the neurula stage, the gene expressions were mainly detected in either paraxial somites or the tail bud. With the development of the embryo, the expression levels further decreased gradually and remained only in some pharyngeal regions or the tail bud at the larva stage. Our results suggest that the Wnt/β-catenin pathway might be involved in amphioxus somite formation and posterior growth, but not in endomesoderm specification.
文摘Objective:To investigate the involvement of reactive oxygen species detoxification system in Anopheles stephensi during Plasmodium berghei midgut invasion.Methods:Eight key reactive oxygen species metabolizing enzymes were cloned and characterized,and their expression was monitored in parasite-infected mosquitoes.Results:Superoxide anion detoxifying superoxide dismutases(Fe/Mn SOD,Cu/Zn SOD 2,Cu/Zn SOD 3A,and Cu/Zn SOD 3B)depicted varied expression patterns.Fe/Mn SOD expression declined,whereas Cu/Zn SOD expression was elevated in the infected mosquitoes.Peroxidases,catalase and glutathione peroxidase showed lack of induction in expression during the Plasmodium berghei infection.Further,expression of thioredoxin reductase increased in the infected mosquitoes,whereas gluthathione S-transferase levels decreased markedly.Conclusions:Detoxification enzymes may play a role in modulating host immunity and parasite transmission.
基金This work was supported by the National Natural Science Foundation of China (No. 39770330).
文摘Objective: To study the molecular mechanism ofdifferent tumorigenicity in nude mice of human leukemiacell lines K562-n and K562. Methods: To analyze the genes differently expressed between K562 and K562-n cells byusing cDNA microarray technique. Results: Among the12800 genes detected, some genes involved in materialmetabolism and material transport were differentlyexpressed between K562-n and K562 cells. These genesinclude homo sapiens placenta-specific ATP-binding cassette transporter gene, dihydrodiol dehydrogenase gene, hepatic dihydrodiol dehydrogenase gene, NAD-dependent methylene tetrahydrofolate dehydrogenase cyclohydrolase,lysophosphatidic acid acyltransferase, alpha gene,argininosuccinate lyase gene, mitochondrial isocitrtatedehydrogenase, adhesion protein SQM1 gene, dimethylarginine dimethylamino-hydrolase gene, M1subunit of ribonucleotide reductase and farnesylpyrophosphate synthetase gene. Conclusion: The hightumorigenicity of K562-n cells is related to the differentexpression of some genes concerned with cell metabolismand material transpoert.
文摘The full-length cDNA encoding the subunits p40 and p35 of human interleukin12(hIL12) were cloned separately by RTPCR, linked together by internal ribosomal entry site (IRES) of encephalomyocarditis virus which initiates capindependent translation to form a dicistronic gene fragment. The dicistronic fragment was placed between the cytomegalovirus (CMV) promoter and SV40 polyA signal to form a dicistronic expression cassette. Subsequently, the dicistronic expression cassette was inserted into E1 region of Ad5 genome in cosmid vector pAx1cw of E1substitution type. By homologous recombination with EcoT22Idigested Ad5 DNATPC in 293 cells, the replicationdeficient recombinant adenoviruses of hIL12 were generated efficiently. After infected with hIL12 recombinant adenoviruses in vitro, 293 cells, human hepatocellular carcinoma cells HepG2, and primary human skin fibroblasts expressed and secreted hIL12 at comparable levels (30～60ng/ 106cells/24hr), which could stimulate the proliferation and IFNγ production of human lymphoblasts. These suggest that the dicistronic adenovirus vector of hIL12 could effectively mediate the expression of bioactive hIL12 and might be used in cancer gene therapy.
文摘Chitin is a common component in the natural diet of many fish.While a range of chitin-degrading chitinases have been identified,it is unclear whether chitin is metabolized in fish.Here we use a combination of chitinase activity assays,transcriptomics,and 16S rRNA bacterial analysis to assess the effect of chitin supplementation on Atlantic salmon gene expression and microbial community composition.Atlantic salmon express multiple genes associated with chitin metabolism.We show that the expression and activity of Atlantic salmon chitinases are not affected by the addition of dietary chitin,but we demonstrate an association between gut microbial composition,chitinase activity in the gut,and host chitinase expression.These results contribute to a greater understanding of chitin metabolism in fish.
基金Supported by the National Natural Science Foundation of China(No.81160528)the Governor Foundation of Guizhou Province(No.2006-07)Administration of Traditional Chinese Medicine of Guizhou Province Foundation(No.2009-79)
文摘Objective： To evaluate the effects and possible mechanisms of rutaecarpine on angiotensin Ⅱ （Ang Ⅱ ）-induced proliferation in cultured rat vascular smooth muscle cells （VSMCs）. Methods： VSMCs were isolated from Male Sprague-Dawley rat aorta, and cultured by enzymic dispersion method. Experiments were performed with cells from passages 3-8. The cultured VSMCs were randomly divided into control, model （Ang Ⅱ 0.1 μ moVL）, and rutaecarpine （0.3-3.0μmol/L） groups. VMSC proliferation was induced by Ang Ⅱ, and was evaluated by the 3-（4, 5-dimethylthiazol-2-yl）-2, 5-diphenyltetrazolium bromide assay and cell counting. To examine the mechanisms involved in anti-proliferative effects of rutaecarpine, nitric oxide （NO） levels and NO synthetase （NOS） activity were determined. Expressions of VSMC proliferation-related genes including endothelial nitric oxide synthase （eNOS）, and c-myc hypertension related gene-1 （HRG-1） were determined by real-time reverse chain reaction （RT-PCR）. Results： Rutaecarpine （0.3-3.0μmol/l_） inhibited Ang R-induced VSMC proliferation and the best effects were achieved at 3.0 μmol/L. The Ang Ⅱ-induced decreases in cellular NO contents and NOS activities were antagonized by rutaecarpine （P〈0.05）. Ang Ⅱ administration suppressed the expressions of eNOS and HRG-1, while increased c-myc expression （P〈0.05）. All these effects were attenuated by 3.0μmol/L rutaecarpine （P〈0.05）. Conclusion： Rutaecarpine is effective against Ang Ⅱ-induced rat VSMC proliferation, and this effect is due, at least in part, to NO production and the modulation of VMSC proliferation-related gene expressions.
基金This research was supported by grants from the Na- tional Natural Science Foundation of China （Grant Nos. 31272505, 31172269 and 31360586）.
文摘Class B scavenger receptors （SR-Bs） are cell surface glycoproteins involved in various physiological processes in vivo, including the transport and metabolism of lipids, binding and phagoeytosis of xenobiotics, and signaling. But little information is available about silkworm SR-Bs; it is necessary to study these SR-Bs for revealing their function. In this study, we cloned the full-length coding sequence of BrnSCRBQ4, a SR-B gene from the silkworm Bombyx mori L. We found that the BmSCRBQ4 gene consists of nine exons and eight introns, with an open reading frame of 1371 bp encoding 456 amino acids. Gene expression studies determined that BmSCRBQ4 messenger RNA （mRNA） was expressed in unfertilized eggs, during embryonic development and throughout the majority of the larval period. Expression of mRNA was detected in the mid gut, middle silk gland, posterior silk gland, head, integumentum, fat body, testes and the ovaries of the larval B. mori Dazao strain, as well as in the silkworm cell lines BmN and BmE. Protein expression studies found BmSCRBQ4 protein was expressed only in the testes, fat body and middle silk gland of larvae, as well as in the silkworm cell lines BmN and BmE. The BmSCRBQ4 protein showed variability in banding patterns in different tissues and cells when analyzed by Western blotting. Immunohistochemical staining showed that the BmSCRBQ4 protein localizes to the constitutive membranes or cellular membranes of these tissues. These results indicated that BmSCRBQ4 gene may play some physiologically relevant roles at the cell surface in each tissue.
基金supported by grants from the National Natural Science Foundation of China(Grant No.81772614)the National Key R&D Program of China(Grant No.2017YFA0106700)+2 种基金the Program for Guangdong Introducing Innovative and Entrepreneurial Teams(Grant No.2017ZT07S096)the Zhejiang Qianjiang Talent Project(Grant No.QJD1602025)the Guangdong Basic and Applied Basic Research Foundation(Grant No.2021B1515020108),China.
文摘Immunotherapy is a promising cancer treatment method;however,only a few patients benefit from it.The development of new immunotherapy strategies and effective biomarkers of response and resistance is urgently needed.Recently,high-throughput bulk and single-cell gene expression profling technologies have generated valuable resources.However,these resources are not well organized and systematic analysis is difficult.Here,we present TIGER,a tumor immunotherapy gene expression resource,which contains bulk transcriptome data of 1508 tumor samples with clinical immunotherapy outcomes and 11,057 tumor/normal samples without clinical immunotherapy outcomes,as well as single-cell transcriptome data of 2,116,945 immune cells from 655 samples.TIGER provides many useful modules for analyzing collected and user-provided data.Using the resource in TIGER,we identified a tumor-enriched subset of CD4^(+)T cells.Patients with melanoma with a higher signature score of this subset have a significantly better response and survival under immunotherapy.We believe that TIGER will be helpful in understanding anti-tumor immunity mechanisms and discovering effective biomarkers.
