BACKGROUND Cyclin-dependent kinase 9(CDK9)expression and autophagy in colorectal cancer(CRC)tissues has not been widely studied.CDK9,a key regulator of transcription,may influence the occurrence and progression of CRC...BACKGROUND Cyclin-dependent kinase 9(CDK9)expression and autophagy in colorectal cancer(CRC)tissues has not been widely studied.CDK9,a key regulator of transcription,may influence the occurrence and progression of CRC.The expression of auto-phagy-related genes BECN1 and drug resistance factor ABCG2 may also play a role in CRC.Under normal physiological conditions,autophagy can inhibit tumorigenesis,but once a tumor forms,autophagy may promote tumor growth.Therefore,understanding the relationship between autophagy and cancer,partic-ularly how autophagy promotes tumor growth after its formation,is a key motivation for this research.AIM To investigate the relationship between CDK9 expression and autophagy in CRC,assess differences in autophagy between left and right colon cancer,and analyze the associations of autophagy-related genes with clinical features and prognosis.METHODS We collected tumor tissues and paracarcinoma tissues from colon cancer patients with liver metastasis to observe the level of autophagy in tissues with high levels of CDK9 and low levels of CDK9.We also collected primary tissue from left and right colon cancer patients with liver metastasis to compare the autophagy levels and the expression of BECN1 and ABCG2 in the tumor and paracarcinoma tissues.RESULTS The incidence of autophagy and the expression of BECN1 and ABCG2 were different in left and right colon cancer,and autophagy might be involved in the occurrence of chemotherapy resistance.Further analysis of the rela-tionship between the expression of autophagy-related genes CDK9,ABCG2,and BECN1 and the clinical features and prognosis of colorectal cancer showed that the high expression of CDK9 indicated a poor prognosis in colorectal cancer.CONCLUSION This study laid the foundation for further research on the combination of CDK9 inhibitors and autophagy inhibitors in the treatment of patients with CRC.展开更多
BACKGROUND The ubiquitin-proteasome pathway(UPP)has been proven to play important roles in cancer.AIM To investigate the prognostic significance of genes involved in the UPP and develop a predictive model for liver ca...BACKGROUND The ubiquitin-proteasome pathway(UPP)has been proven to play important roles in cancer.AIM To investigate the prognostic significance of genes involved in the UPP and develop a predictive model for liver cancer based on the expression of these genes.METHODS In this study,UPP-related E1,E2,E3,deubiquitylating enzyme,and proteasome gene sets were obtained from the Kyoto Encyclopedia of Genes and Genomes(KEGG)database,aiming to screen the prognostic genes using univariate and multivariate regression analysis and develop a prognosis predictive model based RESULTS Five genes(including autophagy related 10,proteasome 20S subunit alpha 8,proteasome 20S subunit beta 2,ubiquitin specific peptidase 17 like family member 2,and ubiquitin specific peptidase 8)were proven significantly correlated with prognosis and used to develop a prognosis predictive model for liver cancer.Among training,validation,and Gene Expression Omnibus sets,the overall survival differed significantly between the high-risk and low-risk groups.The expression of the five genes was significantly associated with immunocyte infiltration,tumor stage,and postoperative recurrence.A total of 111 differentially expressed genes(DEGs)were identified between the high-risk and low-risk groups and they were enriched in 20 and 5 gene ontology and KEGG pathways.Cell division cycle 20,Kelch repeat and BTB domain containing 11,and DDB1 and CUL4 associated factor 4 like 2 were the DEGs in the E3 gene set that correlated with survival.CONCLUSION We have constructed a prognosis predictive model in patients with liver cancer,which contains five genes that associate with immunocyte infiltration,tumor stage,and postoperative recurrence.展开更多
[Objectives]This study was conducted to investigate the effects of Astragalus membranaceus in different groups on energy metabolism and CNTF protein expression in skeletal muscle of exercise-induced fatigue rats.[Meth...[Objectives]This study was conducted to investigate the effects of Astragalus membranaceus in different groups on energy metabolism and CNTF protein expression in skeletal muscle of exercise-induced fatigue rats.[Methods]Thirty-five clean male SD rats were randomly divided into a normal group,and low-,meddle-and high-dose groups of A.membranaceus aqueous solution,with 7 rats in each group.The low-dose,medium-dose and high-dose groups were given by gavage at 0.65,1.3 and 2.6 g/kg,respectively,while the normal group and the model group were given normal food and water.The weight of rats was observed.The contents of serum urea,lactate,muscle glycogen,liver glycogen and CNTF expression were detected.[Results]After modeling,compared with the normal group,the serum lactate and urea contents of rats in the model group significantly increased(P<0.01),while the muscle glycogen content(P<0.01)and liver glycogen content(P<0.05)of the skeletal muscle significantly decreased.Compared with the model group,the low-,meddle-and high-dose groups of A.membranaceus significantly reduced the levels of lactate and urea in serum(P<0.01),while the levels of muscle glycogen and liver glycogen in the skeletal muscle significantly increased(P<0.01,P<0.05).[Conclusions]This study provides a good research foundation for the treatment of exercise-induced fatigue using traditional Chinese herb A.membranaceus in modern clinical practice.展开更多
Adventitious shoot(AS)regeneration is a significant factor in the genetic transformation of horticultural plants.It is also a noteworthy approach to their vegetative propagation.AS regeneration remains highly dependen...Adventitious shoot(AS)regeneration is a significant factor in the genetic transformation of horticultural plants.It is also a noteworthy approach to their vegetative propagation.AS regeneration remains highly dependent on the genotype or maturity of explants.We here found that the AS regeneration abilities of apple leaves were positively correlated with MdAIL5 expression.MdAIL5 overexpression dramatically increased AS regeneration efficiency.Notably,MdAIL5 overexpression could restore the AS formation ability of explants to a certain extent,which was lost with an increase in maturity.Endogenous hormone detection revealed that MdAIL5 overexpression changed the contents of auxin,cytokinin(CK),and other hormones in apple leaves.Transcriptome analysis revealed that many genes related to auxin,CK,and brassinolide signaling pathways were significantly and differentially expressed between MdAIL5-overexpressing transgenic apple and wild-type apple plants.Yeast one-hybrid assays,the electrophoretic mobility shift assay,and the dual-luciferase reporter assay revealed that MdAIL5 directly binds to MdARF9 and MdHB14 promoters and positively affects their expression.We here established a model of MdAIL5 regulating AS formation,which acts as a theoretical basis for facilitating genotype-or explant maturity-independent AS regeneration in the future.展开更多
Assembly geometric error as a part of the machine tool system errors has a significant influence on the machining accuracy of the multi-axis machine tool.And it cannot be eliminated due to the error propagation of com...Assembly geometric error as a part of the machine tool system errors has a significant influence on the machining accuracy of the multi-axis machine tool.And it cannot be eliminated due to the error propagation of components in the assembly process,which is generally non-uniformly distributed in the whole working space.A comprehensive expression model for assembly geometric error is greatly helpful for machining quality control of machine tools to meet the demand for machining accuracy in practice.However,the expression ranges based on the standard quasistatic expression model for assembly geometric errors are far less than those needed in the whole working space of the multi-axis machine tool.To address this issue,a modeling methodology based on the Jacobian-Torsor model is proposed to describe the spatially distributed geometric errors.Firstly,an improved kinematic Jacobian-Torsor model is developed to describe the relative movements such as translation and rotation motion between assembly bodies,respectively.Furthermore,based on the proposed kinematic Jacobian-Torsor model,a spatial expression of geometric errors for the multi-axis machine tool is given.And simulation and experimental verification are taken with the investigation of the spatial distribution of geometric errors on five four-axis machine tools.The results validate the effectiveness of the proposed kinematic Jacobian-Torsor model in dealing with the spatial expression of assembly geometric errors.展开更多
The functional properties of endogenous Schwann cells(SCs)during nerve repair are dynamic.Optimizing the functional properties of SCs at different stages of nerve repair may have therapeutic benefit in improving the r...The functional properties of endogenous Schwann cells(SCs)during nerve repair are dynamic.Optimizing the functional properties of SCs at different stages of nerve repair may have therapeutic benefit in improving the repair of damaged nerves.Previous studies showed that miR-221-3p promotes the proliferation and migration of SCs,and miR-338-3p promotes the myelination of SCs.In this study,we established rat models of sciatic nerve injury by bridging the transected sciatic nerve with a silicone tube.We injected a miR-221 lentiviral vector system together with a doxycycline-inducible Tet-On miR-338 lentiviral vector system into the cavity of nerve conduits of nerve stumps to sequentially regulate the biological function of endogenous SCs at different stages of nerve regeneration.We found that the biological function of SCs was sequentially regulated,the diameter and density of myelinated axons were increased,the expression levels of NF200 and myelin basic protein were increased,and the function of injured peripheral nerve was improved using this system.miRNA Target Prediction Database prediction,Nanopore whole transcriptome sequencing,quantitative PCR,and dual luciferase reporter gene assay results predicted and verified Cdkn1b and Nrp1 as target genes of miR-221-3p and miR-338-3p,respectively,and their regulatory effects on SCs were confirmed in vitro.In conclusion,here we established a new method to enhance nerve regeneration through sequential regulation of biological functions of endogenous SCs,which establishes a new concept and model for the treatment of peripheral nerve injury.The findings from this study will provide direct guiding significance for clinical treatment of sciatic nerve injury.展开更多
Recombinant adenovirus serotype 5(Ad5)vector has been widely applied in vaccine development targeting infectious diseases,such as Ebola virus disease and coronavirus disease 2019(COVID-19).However,the high prevalence ...Recombinant adenovirus serotype 5(Ad5)vector has been widely applied in vaccine development targeting infectious diseases,such as Ebola virus disease and coronavirus disease 2019(COVID-19).However,the high prevalence of preexisting anti-vector immunity compromises the immunogenicity of Ad5-based vaccines.Thus,there is a substantial unmet need to minimize preexisting immunity while improving the insert-induced immunity of Ad5 vectors.Herein,we address this need by utilizing biocompatible nanoparticles to modulate Ad5–host interactions.We show that positively charged human serum albumin nanoparticles((+)HSAnp),which are capable of forming a complex with Ad5,significantly increase the transgene expression of Ad5 in both coxsackievirus–adenovirus receptor-positive and-negative cells.Furthermore,in charge-and dose-dependent manners,Ad5/(+)HSAnp complexes achieve robust(up to227-fold higher)and long-term(up to 60 days)transgene expression in the lungs of mice following intranasal instillation.Importantly,in the presence of preexisting anti-Ad5 immunity,complexed Ad5-based Ebola and COVID-19 vaccines significantly enhance antigen-specific humoral response and mucosal immunity.These findings suggest that viral aggregation and charge modification could be leveraged to engineer enhanced viral vectors for vaccines and gene therapies.展开更多
Understanding physiological responses in saline agriculture may facilitate wheat breeding programs.Based on a screening test,the Ningmai-14(NM-14)and Yangmai-23(YM-23)wheat cultivars were selected for further experime...Understanding physiological responses in saline agriculture may facilitate wheat breeding programs.Based on a screening test,the Ningmai-14(NM-14)and Yangmai-23(YM-23)wheat cultivars were selected for further experiments to understand the underlying salinity tolerance mechanism.This study investigated the effects of five salinity levels such as Control(CK)=0(without NaCl stress),S1=0.20%,S2=0.25%,S3=0.30%and S4=0.35%of NaCl concentrations of soil on wheat plants.The results showed that increased salinity concentration reduced the growth and yield of wheat cultivars(NM-14 and YM-23).However,YM-23(12.7%)yielded more than NM-14 at maximum salinity stress.The higher salinity(S4)increased the concentration of Na^(+)(4.3 to 5.8-fold)and P contents(2.5 to 2.2-fold),while reducing the average concentrations of K^(+),Cu,and K^(+)/Na^(+)ratio.The higher salinity(S4)reduced the spikelet length by 21.35%(followed by grain spike−1),and the starch content by 18.81%.In the YM-23 cultivar,higher salinity increased superoxide dismutase(SOD),total antioxidant capacity(TAC),and amylase.Compared to NM-14,induced expression of TaYUC2,6,and TaGA13ox,20ox genes were recorded in YM-23.Similarly,in YM-23 the stress-specific genes such as TaHSP70,90 were enhanced whereas,TaSOS1,2 were suppressed.Overall,our study revealed that salt tolerant cultivars modulate hormonal and antioxidant activities,thus maintaining high growth.展开更多
Objective:To study the inhibitory effect of siRNA mediated by expression vectors on Skp2 expression in human chondrocytes.Method:Three Skp2 sequences siRNA-59,siRNA-318 and siRNA-504 were designed as target sites usin...Objective:To study the inhibitory effect of siRNA mediated by expression vectors on Skp2 expression in human chondrocytes.Method:Three Skp2 sequences siRNA-59,siRNA-318 and siRNA-504 were designed as target sites using online siRNA design tools.Skp2 siRNA expression vectors were successfully constructed in vitro by gene recombination technology,and the influence of recombinant plasmid transfection on Skp2 mRNA expression was detected.DNA electrophoresis was used to verify the results.Results:The sequence of Skp2 interference was correct by sequence analysis.The expression of Skp2 mRNA in siRNA-59,siRNA-318,siRNA-504 transfection group was significantly lower than that in no-load group and NC group(P<0.05),the inhibition rates of Skp2 mRNA in siRNA-59,siRNA-318 and siRNA-504 were respectively 60%,41%and 64%,and the siRNA-504 transfection group had the highest inhibition rate.Conclusion:The siRNA eukaryotic expression vector of Skp2 gene was constructed successfully which effectively inhibit Skp2 mRNA expression in human chondrocytes cell,and can provide strong evidence for the treatment of osteoarthritis.展开更多
Photodamage continues to threaten human skin health despite worldwide sun awareness campaigns and the widespread use of sunscreens. To date, extensive research is lacking into the effects of sun avoidance and solar sp...Photodamage continues to threaten human skin health despite worldwide sun awareness campaigns and the widespread use of sunscreens. To date, extensive research is lacking into the effects of sun avoidance and solar specific skincare regimens on gene expression changes and DNA repair activity. We have previously reported that photoprotection and photorepair formulations which minimize the harmful effects of ultraviolet, visible light and near-infrared radiation can provide photoprotection, anti-photoaging benefits and rejuvenating effects optically, clinically and genetically. To investigate gene expression changes, specifically antioxidant and DNA repair effects following the use of topical photoprotection and photorepair formulations (The Essential Six, RATIONALE, Victoria, Australia), we used epidermal keratinocytes and dermal fibroblasts derived from a 3-dimensional reconstructed human skin model, and assessed upregulation of SOD2 and HPRT1. Gene expression was assessed via the Genemarkers Standard Skin Panel and quantitative real-time PCR exploration. Tissues were inoculated with solar specific topical formulations, then collected after 24 hours following application of photoprotection formulations and 16 hours following photorepair formulations. The quantitative real-time PCR revealed that, in comparison to the control, the genes encoding SOD2 and HPRT1 have been significantly up-regulated following usage of the photoprotection formulations, 1.86, and 1.41, respectively. SOD2 and HPRT1 were up-regulated following use of the photorepair formulations, 2.15, and 1.28, respectively. We were able to substantiate that the photo protection and photorepair formulations upregulated genes involved in antioxidant and DNA repair mechanisms in a 3-dimensional reconstructed human skin model, suggesting a promising anti-photoaging skin regimen. .展开更多
The effects of Ce (Ⅳ) on callus growth, anthocyanin content, and expression of anthocyanin biosynthetic genes in callus suspension cultures of Solanum tuberosum cv. Chieftain were studied by the measurement of fres...The effects of Ce (Ⅳ) on callus growth, anthocyanin content, and expression of anthocyanin biosynthetic genes in callus suspension cultures of Solanum tuberosum cv. Chieftain were studied by the measurement of fresh weight, spectrophotometric assays, and semiquantitative RT-PCR. The results indicate that 0.1 mmol·L^- 1 Ce ( Ⅳ ) can promote callus growth, increase the accumulation of anthocyanins, and enhance the expression of five anthocyanin biosynthetic genes ( CHS, F3H, F3'5'H, DFR, and 3 GT) most efficiently. At high concentrations of 1 mmol·L^- 1, Ce (Ⅳ) partially inhibits callus growth and at 2 mmol· L^-1 eventually lends to cell death. The results show that Ce( Ⅳ ) can induce the expression of anthocyanin biosynthetic genes to produce and accumulate anthocyanins and increase the yield of anthocyanins.展开更多
Selenium (Se) is an essential trace element. Autoimmune thyroid diseases (AITD) are destructive inflammatory or anti-receptor autoimmune diseases characterized by reactivity to self-thyroid antigens. However, the ...Selenium (Se) is an essential trace element. Autoimmune thyroid diseases (AITD) are destructive inflammatory or anti-receptor autoimmune diseases characterized by reactivity to self-thyroid antigens. However, the effects of Se on the cytokines in AITD are still unclear. So we researched the role of Selenium (Se) and Thl/Th2 cytokine productions in the pathogenesis of autoimmune thyroid diseases (AITD).展开更多
In the screening tests of drugs for silicosis in our laboratory, we found that TT, a type of alkaloid isolated from Stephania tetrandra, could inhibit the development of experimental silicosis of rats and the synthesi...In the screening tests of drugs for silicosis in our laboratory, we found that TT, a type of alkaloid isolated from Stephania tetrandra, could inhibit the development of experimental silicosis of rats and the synthesis of collagen in rat lung. Chest X-rays of silicotic patients treaied with TT for 1-3 years showed obvious changes. The silicotic nodules became smallel and shadows became clearer. PVNO was proved to have anti-silicotic effect on animal and clinically. This presentation reports the effect of them on collagen mRNA.Dot blot results showed that 1 (Ⅰ) and 1 (Ⅲ) mRNA levels increased significantly at 60 and 120 days after the rats were exposed to silica dust. The mRNA levels went down at 1 and 3 months after treated by TT and PVNO. In situ hybridization observation revealed that the silver grains of Type Ⅰand Type Ⅲ collagen were scattered within the fibroblasts in cellular nodules and in thickened interstitium of silicosis tissue. The amounts of mRNA silver grains decreased in the lung tissue treated by TT and PVNO. It was suggested that TT and PVNO may inhibil the gene expression of collagen during silicosis展开更多
Antimicrobial peptides (AMPs) are making headlines in science because they demonstrate superior microbicidal characteristics compared to synthetic and semi-synthetic antibiotics.
This paper is aimed to study the effect of ADL on expression of ~z-AR and Mz-AchR in myocardial cells of rats exposed to microwave radiation. Immunohistochemistry, Western blot and image analysis were used to detect t...This paper is aimed to study the effect of ADL on expression of ~z-AR and Mz-AchR in myocardial cells of rats exposed to microwave radiation. Immunohistochemistry, Western blot and image analysis were used to detect the expression of ~I-AR and Mz-AchR in myocardial cells at 7 and 14 d after microwave exposure. The results show that the expression level was higher in microwave exposure group and 0.75 g/(kg.d) ADL group than in sham operation group and significantly lower in 1.5 and 3.0 g/(kg.d) ADL groups than in microwave group. So we have a conclusion that the expression of I^z-AR and Mz-AchR is down-regulated in myocardial cells of rats exposed to microwave radiation. ADL can protect rats against microwave-induced heart tissue injury.展开更多
Silicosis is one of the most serious occupational diseases in China and dates back to centuries ago. In this study, we successfully established a rat model of silicosis by intratracheal silica injection for 28 days an...Silicosis is one of the most serious occupational diseases in China and dates back to centuries ago. In this study, we successfully established a rat model of silicosis by intratracheal silica injection for 28 days and determined hydroxyproline levels to evaluate collagen metabolism in lung homogenates. Oxidative stress status was evaluated by detecting catalase and glutathione peroxidase activities.展开更多
To investigate the mechanisms of microwave induced pacemaker cell injuries, Wistar rats and the primary pacemaker cells of newborn Wistar rats were exposed to microwave at average power density of 50 mW/cm2. Slower sp...To investigate the mechanisms of microwave induced pacemaker cell injuries, Wistar rats and the primary pacemaker cells of newborn Wistar rats were exposed to microwave at average power density of 50 mW/cm2. Slower spontaneous beating rate, intercellular Ca2+ aggregation and cell membrane perforation were detected immediately after the exposure. Moreover, hyperpolarizationactivated cyclic nucleotide-gated cation channel 4 (HCN4) was down-regulated immediately after the exposure and up-regulated at 12 h after the exposure. In the sinoatrial node (SAN) of the rats,展开更多
The hedgehog (Hh) signaling pathway plays an essential role in the embryonic development and homeostasis of diverse adult tissues, and its deregulation has been implicated in the tumorigenesis and metastasis of vari...The hedgehog (Hh) signaling pathway plays an essential role in the embryonic development and homeostasis of diverse adult tissues, and its deregulation has been implicated in the tumorigenesis and metastasis of various malignancies including breast cancer. Aberrant activation of the Hh pathway includes the following mechanisms: (I) Hh ligand-independent mechanism - Loss of function mutations in the Hh receptor Patched 1 (PTCH1) or gain of function mutations in the Smoothened (SMO) lead to constitutive activation of this pathway; (II) Autocrine signaling- Ith ligand produced by tumor cells stimulates the Hh signaling in tumor cells; (III) Paracrine signaling - tumor cell produced-Hh ligand activates stromal and endothelial cells that produce growth factors in microenvironment for supporting tumor growth and survival; and (IV) Reverse paracrine signaling - Hh ligand produced by stromal cells support tumor growth and survival. Upon the pathway activation, the Gli transcription factors, effectors of the Hh signaling, activate or inhibit transcription by binding to their responsive genes and interacting with the transcriptional complex. The Gli transcription factor family includes Glil, Gli2, and Gli3 (1). Glil is a transcriptional activator whose expression has been recognized as an activation state of the Hh signaling pathway, Gli2 is either an activator or repressor, and Gli3 is a strong repressor of transcriptional activities. To date, a ligand-dependent autocrine model of activating the Hh signaling has been described in breast cancer, and both an autocrine and paracrine mechanisms in colorectal cancer, pancreatic cancer and prostate cancer (2,3). Notably, a ligand-independent mechanism (mutationsin PTCHI and SMO) of the signaling has been well demonstrated in basal cell carcinoma and medulloblastoma (4,5).展开更多
Background The cellular repressor of ElA-stimulated genes(CREG) is a secreted glycoprotein that inhibits cell proliferation and/or enhances differentiation.CREG is widely expressed in adult tissues such as the brain,h...Background The cellular repressor of ElA-stimulated genes(CREG) is a secreted glycoprotein that inhibits cell proliferation and/or enhances differentiation.CREG is widely expressed in adult tissues such as the brain,heart, lungs,liver,intestines and kidneys in mice.We investigated the level of CREG expression during mouse embryogenesis and its distribution at 18.5 days post coitus(dpc).Methods Immunohistochemical staining with diaminobenzidine,western blotting and reverse transcription-polymerase chain reaction were used.Results CREG expression was rst detected in mouse embryos at 4.5 dpc.It was expressed at almost all stages up to 18.5 dpc.The level of CREG was found to increase gradually and was highest at 18.5 dpc.Western blotting showed that the CREG protein was expressed at higher levels in the brain,heart,intestines and kidneys than in the lungs and liver at 18.5 dpc.In 9.5 dpc embryos,CREG was expressed only in the endothelial cells of blood vessels,after the vascular lumen had formed.With advanced differentiation, vascular smooth muscle cells developed in the embryonic vascular structures;the expression of smooth muscle a-actin protein and CREG were positive and increased gradually in 10.5 dpc embryonic vessels.CREG expression in the embryonic blood vessels peaked at 15.5 dpc and was reduced slightly at 18.5 dpc.Conclusions These results indicate that CREG is expressed during mouse embryogenesis and might participate in the differentiation of these organs during embryogenesis.展开更多
BACKGROUND: Hypoxia inducible factor-1 alpha (HIF-1 (x) and erythropoietin(EPO), possessing neuroprotective effect in the cerebral ischemia, might play an important role in the formation of cerebral ischemic tol...BACKGROUND: Hypoxia inducible factor-1 alpha (HIF-1 (x) and erythropoietin(EPO), possessing neuroprotective effect in the cerebral ischemia, might play an important role in the formation of cerebral ischemic tolerance (IT). OBJECTIVE:To observe the neuroprotective effect of cerebral ischemic preconditioning(IPC) of rats, and the expression and mechanism of HIF-1α and target gene erythropoietin in the brain tissue following the formation of cerebral IT. DESIGN : A randomized and controlled observation SETTING: Department of Neurology, the Affiliated Hospital of Medical College, Qingdao University MATERIALS: Totally 84 enrolled adult healthy male Wistar rats of clean grade, weighing 250 to 300 g, were provided by the Animal Experimental Department, Tongji Medical College of Huazhong University of Science and Technology. Ready-to-use SABC reagent kit and rabbit anti-rat HIF-1α monoclonal antibody were purchased from Boshide Bioengineering Co.Ltd (Wuhan); Rabbit anti-rat EPO monoclonal antibody was purchased from Santa Cruz Company (USA). METHODS: This experiment was carried out in the Department of Anatomy, Medical College, Qingdao University during March 2005 to March 2006. ① The 84 rats were divided into 3 groups by a lot: IPC group (n=40), sham-operation group (n=40) and control group (n=4). In the IPC group, middle cerebral artery was occluded for 2 hours respectively on the 1^st, 3^rd, 7^th, 14^th and 21^st days of the reperfusion following 10-minute preischemia was made using a modified middle cerebral artery second suture method from Zea-Longa. The rats were sacrificed 22 hours after reperfusion in the end of middle cerebral artery occlusion (MCAO). That was to say, after 10-minute preischemia, suture was exited to the extemal carotid artery and embedded subcutaneously. Middle cerebral artery was occluded again to form the second reperfusion at the set time point after reperfusion. Twenty-two hours later, rats were sacrificed; In the sham-operation group,the preischemia was substituted by sham-operation(only common carotid artery and crotch were exposed, and MCAO by suture was omitted), and the other procedures were the same as those in the IPC group. In the control group, rats were given sham-operation twice at an interval of one day, and they were sacrificed 24 hours after the second sham-operation. ② Brain tissue was taken from the rats in each group. Cerebral infarction area of each layer was measured with TTC staining, and total cerebral infarction volume (The total cerebral infarction area of each layerxinterspace ) was calculated. After brain tissue was stained by haematoxylin-esoin (HE), the form of nerve cells was observed under an optical microscope, and the expressions of HIF-1α(and EPO protein in the brain tissue were detected with immunohistochemical method. MAIN OUTCOME MEASURES: ①Cerebral infarction volume;②form of nerve cell; ③ the expression of HIF-1α and EPO protein in the brain tissue. RESULTS:Totally 84 rats were enrolled in the experiment. The dead rats were randomly supplied during the experiment, and finally 84 rats entered the stage of result analysis. ① Detection of cerebral infarction volume of rats in each group: Cerebral infarction volume in the IPC group was significantly smaller than that in the sham-operation group on the 1^st, 3^rd and 7^th days after reperfusion respectively [(161.2±6.9) mm^3 vs (219.9±11.2) mm^3, (134.9±9.0) mm^3 vs (218.6±13.0) mm^3, (142.9±13.7) mm^3 vs (221.3±14.2) mm^3, t=-8.924, 10.587,7.947, P〈 0.01]. ② Observation of nerve cell form of brain tissue: HE staining showed that the ischemic degree, range and cerebral edema degree of IPC group were significantly milder than those of sham-operation group. ③ The expressions of HIF-1α and EPO protein in cerebral cortex and hippocampus : The expression of HIF-1αof IPC group was significantly higher than that of sham-operation group on the 1^st, 3^rd and 7^th days after reperfusion respectively (125.93±3.79 vs 117.65±5.60, 140.63±4.64 vs 119.33±4.26, 131.15±2.74 vs 107.60±3.89, t=2.449, 6.763,9.899,P 〈 0.05-0.01). The expression of EPO of IPC group was significantly higher than that of sham-operation group on the 3^rd and 7^th days after perfusion respectively (141.68±3.29 vs 126.33±4.51, 138.88±2.59 vs 125.58±6.18,t=5.499,3.970, P〈 0.05). CONCLUSION : ①IPC can protect the never cells in rat brain and the best time to onset of cerebral IT induced by IPC is 1 to 7 days after reperfusion. ② Neuroprotective effect of cerebral IT might be related to the expression of HIF-1α and its target gene EPO.展开更多
基金the Science and Technology Development Fund of Tianjin Education Commission for Higher Education,No.2020KJ133Tianjin Key Medical Discipline(Specialty)Construction Project,No.TJYXZDXK-009A.
文摘BACKGROUND Cyclin-dependent kinase 9(CDK9)expression and autophagy in colorectal cancer(CRC)tissues has not been widely studied.CDK9,a key regulator of transcription,may influence the occurrence and progression of CRC.The expression of auto-phagy-related genes BECN1 and drug resistance factor ABCG2 may also play a role in CRC.Under normal physiological conditions,autophagy can inhibit tumorigenesis,but once a tumor forms,autophagy may promote tumor growth.Therefore,understanding the relationship between autophagy and cancer,partic-ularly how autophagy promotes tumor growth after its formation,is a key motivation for this research.AIM To investigate the relationship between CDK9 expression and autophagy in CRC,assess differences in autophagy between left and right colon cancer,and analyze the associations of autophagy-related genes with clinical features and prognosis.METHODS We collected tumor tissues and paracarcinoma tissues from colon cancer patients with liver metastasis to observe the level of autophagy in tissues with high levels of CDK9 and low levels of CDK9.We also collected primary tissue from left and right colon cancer patients with liver metastasis to compare the autophagy levels and the expression of BECN1 and ABCG2 in the tumor and paracarcinoma tissues.RESULTS The incidence of autophagy and the expression of BECN1 and ABCG2 were different in left and right colon cancer,and autophagy might be involved in the occurrence of chemotherapy resistance.Further analysis of the rela-tionship between the expression of autophagy-related genes CDK9,ABCG2,and BECN1 and the clinical features and prognosis of colorectal cancer showed that the high expression of CDK9 indicated a poor prognosis in colorectal cancer.CONCLUSION This study laid the foundation for further research on the combination of CDK9 inhibitors and autophagy inhibitors in the treatment of patients with CRC.
基金the Tianjin Municipal Natural Science Foundation,No.21JCYBJC01110。
文摘BACKGROUND The ubiquitin-proteasome pathway(UPP)has been proven to play important roles in cancer.AIM To investigate the prognostic significance of genes involved in the UPP and develop a predictive model for liver cancer based on the expression of these genes.METHODS In this study,UPP-related E1,E2,E3,deubiquitylating enzyme,and proteasome gene sets were obtained from the Kyoto Encyclopedia of Genes and Genomes(KEGG)database,aiming to screen the prognostic genes using univariate and multivariate regression analysis and develop a prognosis predictive model based RESULTS Five genes(including autophagy related 10,proteasome 20S subunit alpha 8,proteasome 20S subunit beta 2,ubiquitin specific peptidase 17 like family member 2,and ubiquitin specific peptidase 8)were proven significantly correlated with prognosis and used to develop a prognosis predictive model for liver cancer.Among training,validation,and Gene Expression Omnibus sets,the overall survival differed significantly between the high-risk and low-risk groups.The expression of the five genes was significantly associated with immunocyte infiltration,tumor stage,and postoperative recurrence.A total of 111 differentially expressed genes(DEGs)were identified between the high-risk and low-risk groups and they were enriched in 20 and 5 gene ontology and KEGG pathways.Cell division cycle 20,Kelch repeat and BTB domain containing 11,and DDB1 and CUL4 associated factor 4 like 2 were the DEGs in the E3 gene set that correlated with survival.CONCLUSION We have constructed a prognosis predictive model in patients with liver cancer,which contains five genes that associate with immunocyte infiltration,tumor stage,and postoperative recurrence.
基金Supported by Undergraduate Innovation and Entrepreneurship Training Program of Guizhou University of Traditional Chinese Medicine(GZYDCHZ[2019]42)National Key R&D Plan(2019YFC1712500)Guizhou Provincial Science and Technology Planning Project(QKHHBZ[2020]3003).
文摘[Objectives]This study was conducted to investigate the effects of Astragalus membranaceus in different groups on energy metabolism and CNTF protein expression in skeletal muscle of exercise-induced fatigue rats.[Methods]Thirty-five clean male SD rats were randomly divided into a normal group,and low-,meddle-and high-dose groups of A.membranaceus aqueous solution,with 7 rats in each group.The low-dose,medium-dose and high-dose groups were given by gavage at 0.65,1.3 and 2.6 g/kg,respectively,while the normal group and the model group were given normal food and water.The weight of rats was observed.The contents of serum urea,lactate,muscle glycogen,liver glycogen and CNTF expression were detected.[Results]After modeling,compared with the normal group,the serum lactate and urea contents of rats in the model group significantly increased(P<0.01),while the muscle glycogen content(P<0.01)and liver glycogen content(P<0.05)of the skeletal muscle significantly decreased.Compared with the model group,the low-,meddle-and high-dose groups of A.membranaceus significantly reduced the levels of lactate and urea in serum(P<0.01),while the levels of muscle glycogen and liver glycogen in the skeletal muscle significantly increased(P<0.01,P<0.05).[Conclusions]This study provides a good research foundation for the treatment of exercise-induced fatigue using traditional Chinese herb A.membranaceus in modern clinical practice.
基金supported by the China Agriculture Research System(Grant No.CARS-27)the National Natural Science Foundation of China(Grant No.32202463)the Agricultural Science and Technology Innovation Program(Grant No.CAAS-ASTIP-2021-RIP-02).
文摘Adventitious shoot(AS)regeneration is a significant factor in the genetic transformation of horticultural plants.It is also a noteworthy approach to their vegetative propagation.AS regeneration remains highly dependent on the genotype or maturity of explants.We here found that the AS regeneration abilities of apple leaves were positively correlated with MdAIL5 expression.MdAIL5 overexpression dramatically increased AS regeneration efficiency.Notably,MdAIL5 overexpression could restore the AS formation ability of explants to a certain extent,which was lost with an increase in maturity.Endogenous hormone detection revealed that MdAIL5 overexpression changed the contents of auxin,cytokinin(CK),and other hormones in apple leaves.Transcriptome analysis revealed that many genes related to auxin,CK,and brassinolide signaling pathways were significantly and differentially expressed between MdAIL5-overexpressing transgenic apple and wild-type apple plants.Yeast one-hybrid assays,the electrophoretic mobility shift assay,and the dual-luciferase reporter assay revealed that MdAIL5 directly binds to MdARF9 and MdHB14 promoters and positively affects their expression.We here established a model of MdAIL5 regulating AS formation,which acts as a theoretical basis for facilitating genotype-or explant maturity-independent AS regeneration in the future.
基金Supported by National Natural Science Foundation of China (Grant No.51975369)National Key Science and Technology Research Program of China (Grant No.2019ZX04027001)。
文摘Assembly geometric error as a part of the machine tool system errors has a significant influence on the machining accuracy of the multi-axis machine tool.And it cannot be eliminated due to the error propagation of components in the assembly process,which is generally non-uniformly distributed in the whole working space.A comprehensive expression model for assembly geometric error is greatly helpful for machining quality control of machine tools to meet the demand for machining accuracy in practice.However,the expression ranges based on the standard quasistatic expression model for assembly geometric errors are far less than those needed in the whole working space of the multi-axis machine tool.To address this issue,a modeling methodology based on the Jacobian-Torsor model is proposed to describe the spatially distributed geometric errors.Firstly,an improved kinematic Jacobian-Torsor model is developed to describe the relative movements such as translation and rotation motion between assembly bodies,respectively.Furthermore,based on the proposed kinematic Jacobian-Torsor model,a spatial expression of geometric errors for the multi-axis machine tool is given.And simulation and experimental verification are taken with the investigation of the spatial distribution of geometric errors on five four-axis machine tools.The results validate the effectiveness of the proposed kinematic Jacobian-Torsor model in dealing with the spatial expression of assembly geometric errors.
基金supported by the National Natural Science Foundation of China,No.81771351the National Key R&D Program of China,No.2017YFA0105802+1 种基金the Joint Research Fund Liaoning-Shenyang National Laboratory for Materials Science,No.2019JH3/30100022the National Science Foundation for Post-doctoral Scientists of China,No.2018M641732(all to QA and LLW)。
文摘The functional properties of endogenous Schwann cells(SCs)during nerve repair are dynamic.Optimizing the functional properties of SCs at different stages of nerve repair may have therapeutic benefit in improving the repair of damaged nerves.Previous studies showed that miR-221-3p promotes the proliferation and migration of SCs,and miR-338-3p promotes the myelination of SCs.In this study,we established rat models of sciatic nerve injury by bridging the transected sciatic nerve with a silicone tube.We injected a miR-221 lentiviral vector system together with a doxycycline-inducible Tet-On miR-338 lentiviral vector system into the cavity of nerve conduits of nerve stumps to sequentially regulate the biological function of endogenous SCs at different stages of nerve regeneration.We found that the biological function of SCs was sequentially regulated,the diameter and density of myelinated axons were increased,the expression levels of NF200 and myelin basic protein were increased,and the function of injured peripheral nerve was improved using this system.miRNA Target Prediction Database prediction,Nanopore whole transcriptome sequencing,quantitative PCR,and dual luciferase reporter gene assay results predicted and verified Cdkn1b and Nrp1 as target genes of miR-221-3p and miR-338-3p,respectively,and their regulatory effects on SCs were confirmed in vitro.In conclusion,here we established a new method to enhance nerve regeneration through sequential regulation of biological functions of endogenous SCs,which establishes a new concept and model for the treatment of peripheral nerve injury.The findings from this study will provide direct guiding significance for clinical treatment of sciatic nerve injury.
基金supported in part by the grant from National Natural Science Foundation of China(82171818,81703048,82041019,and 82101919)the grant from Defense Industrial Technology Development Program of China(JCKY2020802B001)Beijing Municipal Science and Technology Commission(Z201100005420024)。
文摘Recombinant adenovirus serotype 5(Ad5)vector has been widely applied in vaccine development targeting infectious diseases,such as Ebola virus disease and coronavirus disease 2019(COVID-19).However,the high prevalence of preexisting anti-vector immunity compromises the immunogenicity of Ad5-based vaccines.Thus,there is a substantial unmet need to minimize preexisting immunity while improving the insert-induced immunity of Ad5 vectors.Herein,we address this need by utilizing biocompatible nanoparticles to modulate Ad5–host interactions.We show that positively charged human serum albumin nanoparticles((+)HSAnp),which are capable of forming a complex with Ad5,significantly increase the transgene expression of Ad5 in both coxsackievirus–adenovirus receptor-positive and-negative cells.Furthermore,in charge-and dose-dependent manners,Ad5/(+)HSAnp complexes achieve robust(up to227-fold higher)and long-term(up to 60 days)transgene expression in the lungs of mice following intranasal instillation.Importantly,in the presence of preexisting anti-Ad5 immunity,complexed Ad5-based Ebola and COVID-19 vaccines significantly enhance antigen-specific humoral response and mucosal immunity.These findings suggest that viral aggregation and charge modification could be leveraged to engineer enhanced viral vectors for vaccines and gene therapies.
基金the National Natural Science Foundation of China(32101817)Jiangsu Agriculture Science and this work was funded by the National Natural Science Foundation of China(32101817)+3 种基金Jiangsu Agriculture Science and Technology Innovation Fund(CX(21)3111)the Natural Science Foundation of the Jiangsu Higher Education Institutions(21KJD210001)the Scientific and Technological Innovation Fund of Carbon Emissions Peak and Neutrality of Jiangsu Provincial Department of Science and Technology(BE2022304)the project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD)for their financial support.
文摘Understanding physiological responses in saline agriculture may facilitate wheat breeding programs.Based on a screening test,the Ningmai-14(NM-14)and Yangmai-23(YM-23)wheat cultivars were selected for further experiments to understand the underlying salinity tolerance mechanism.This study investigated the effects of five salinity levels such as Control(CK)=0(without NaCl stress),S1=0.20%,S2=0.25%,S3=0.30%and S4=0.35%of NaCl concentrations of soil on wheat plants.The results showed that increased salinity concentration reduced the growth and yield of wheat cultivars(NM-14 and YM-23).However,YM-23(12.7%)yielded more than NM-14 at maximum salinity stress.The higher salinity(S4)increased the concentration of Na^(+)(4.3 to 5.8-fold)and P contents(2.5 to 2.2-fold),while reducing the average concentrations of K^(+),Cu,and K^(+)/Na^(+)ratio.The higher salinity(S4)reduced the spikelet length by 21.35%(followed by grain spike−1),and the starch content by 18.81%.In the YM-23 cultivar,higher salinity increased superoxide dismutase(SOD),total antioxidant capacity(TAC),and amylase.Compared to NM-14,induced expression of TaYUC2,6,and TaGA13ox,20ox genes were recorded in YM-23.Similarly,in YM-23 the stress-specific genes such as TaHSP70,90 were enhanced whereas,TaSOS1,2 were suppressed.Overall,our study revealed that salt tolerant cultivars modulate hormonal and antioxidant activities,thus maintaining high growth.
基金National Natural Science Foundation of China(82060875)National Natural Science Foundation of China(82160912)Guangxi Natural Science Foundation(2022JJA141229)。
文摘Objective:To study the inhibitory effect of siRNA mediated by expression vectors on Skp2 expression in human chondrocytes.Method:Three Skp2 sequences siRNA-59,siRNA-318 and siRNA-504 were designed as target sites using online siRNA design tools.Skp2 siRNA expression vectors were successfully constructed in vitro by gene recombination technology,and the influence of recombinant plasmid transfection on Skp2 mRNA expression was detected.DNA electrophoresis was used to verify the results.Results:The sequence of Skp2 interference was correct by sequence analysis.The expression of Skp2 mRNA in siRNA-59,siRNA-318,siRNA-504 transfection group was significantly lower than that in no-load group and NC group(P<0.05),the inhibition rates of Skp2 mRNA in siRNA-59,siRNA-318 and siRNA-504 were respectively 60%,41%and 64%,and the siRNA-504 transfection group had the highest inhibition rate.Conclusion:The siRNA eukaryotic expression vector of Skp2 gene was constructed successfully which effectively inhibit Skp2 mRNA expression in human chondrocytes cell,and can provide strong evidence for the treatment of osteoarthritis.
文摘Photodamage continues to threaten human skin health despite worldwide sun awareness campaigns and the widespread use of sunscreens. To date, extensive research is lacking into the effects of sun avoidance and solar specific skincare regimens on gene expression changes and DNA repair activity. We have previously reported that photoprotection and photorepair formulations which minimize the harmful effects of ultraviolet, visible light and near-infrared radiation can provide photoprotection, anti-photoaging benefits and rejuvenating effects optically, clinically and genetically. To investigate gene expression changes, specifically antioxidant and DNA repair effects following the use of topical photoprotection and photorepair formulations (The Essential Six, RATIONALE, Victoria, Australia), we used epidermal keratinocytes and dermal fibroblasts derived from a 3-dimensional reconstructed human skin model, and assessed upregulation of SOD2 and HPRT1. Gene expression was assessed via the Genemarkers Standard Skin Panel and quantitative real-time PCR exploration. Tissues were inoculated with solar specific topical formulations, then collected after 24 hours following application of photoprotection formulations and 16 hours following photorepair formulations. The quantitative real-time PCR revealed that, in comparison to the control, the genes encoding SOD2 and HPRT1 have been significantly up-regulated following usage of the photoprotection formulations, 1.86, and 1.41, respectively. SOD2 and HPRT1 were up-regulated following use of the photorepair formulations, 2.15, and 1.28, respectively. We were able to substantiate that the photo protection and photorepair formulations upregulated genes involved in antioxidant and DNA repair mechanisms in a 3-dimensional reconstructed human skin model, suggesting a promising anti-photoaging skin regimen. .
基金Project Supported bythe International Cooperation Research of Jiangsu Province (BZ2003041)
文摘The effects of Ce (Ⅳ) on callus growth, anthocyanin content, and expression of anthocyanin biosynthetic genes in callus suspension cultures of Solanum tuberosum cv. Chieftain were studied by the measurement of fresh weight, spectrophotometric assays, and semiquantitative RT-PCR. The results indicate that 0.1 mmol·L^- 1 Ce ( Ⅳ ) can promote callus growth, increase the accumulation of anthocyanins, and enhance the expression of five anthocyanin biosynthetic genes ( CHS, F3H, F3'5'H, DFR, and 3 GT) most efficiently. At high concentrations of 1 mmol·L^- 1, Ce (Ⅳ) partially inhibits callus growth and at 2 mmol· L^-1 eventually lends to cell death. The results show that Ce( Ⅳ ) can induce the expression of anthocyanin biosynthetic genes to produce and accumulate anthocyanins and increase the yield of anthocyanins.
基金supported by three programs from the National Natural Science Foundation of China (NSFC) (The experimental study on the effect of trace elements iodine and selenium on the autoimmune thyroid disease (No.30571564)The cross-sectional investigation on hypothyroidism induced by excess iodine intake and the experimental research on pathogenesy (No.30972465)The change of thyroid pathology and the levels of T3,T4 in SePP1,GPX3 knock out mice (No.30810103004)
文摘Selenium (Se) is an essential trace element. Autoimmune thyroid diseases (AITD) are destructive inflammatory or anti-receptor autoimmune diseases characterized by reactivity to self-thyroid antigens. However, the effects of Se on the cytokines in AITD are still unclear. So we researched the role of Selenium (Se) and Thl/Th2 cytokine productions in the pathogenesis of autoimmune thyroid diseases (AITD).
文摘In the screening tests of drugs for silicosis in our laboratory, we found that TT, a type of alkaloid isolated from Stephania tetrandra, could inhibit the development of experimental silicosis of rats and the synthesis of collagen in rat lung. Chest X-rays of silicotic patients treaied with TT for 1-3 years showed obvious changes. The silicotic nodules became smallel and shadows became clearer. PVNO was proved to have anti-silicotic effect on animal and clinically. This presentation reports the effect of them on collagen mRNA.Dot blot results showed that 1 (Ⅰ) and 1 (Ⅲ) mRNA levels increased significantly at 60 and 120 days after the rats were exposed to silica dust. The mRNA levels went down at 1 and 3 months after treated by TT and PVNO. In situ hybridization observation revealed that the silver grains of Type Ⅰand Type Ⅲ collagen were scattered within the fibroblasts in cellular nodules and in thickened interstitium of silicosis tissue. The amounts of mRNA silver grains decreased in the lung tissue treated by TT and PVNO. It was suggested that TT and PVNO may inhibil the gene expression of collagen during silicosis
基金supported by grants from the Project of Jilin province Technology Platform(NO.20070210)the Young Research Fond of Jilin University
文摘Antimicrobial peptides (AMPs) are making headlines in science because they demonstrate superior microbicidal characteristics compared to synthetic and semi-synthetic antibiotics.
文摘This paper is aimed to study the effect of ADL on expression of ~z-AR and Mz-AchR in myocardial cells of rats exposed to microwave radiation. Immunohistochemistry, Western blot and image analysis were used to detect the expression of ~I-AR and Mz-AchR in myocardial cells at 7 and 14 d after microwave exposure. The results show that the expression level was higher in microwave exposure group and 0.75 g/(kg.d) ADL group than in sham operation group and significantly lower in 1.5 and 3.0 g/(kg.d) ADL groups than in microwave group. So we have a conclusion that the expression of I^z-AR and Mz-AchR is down-regulated in myocardial cells of rats exposed to microwave radiation. ADL can protect rats against microwave-induced heart tissue injury.
基金supported by the Applied Basic Research Program at Hebei Province in China(Grant No.11966120D)
文摘Silicosis is one of the most serious occupational diseases in China and dates back to centuries ago. In this study, we successfully established a rat model of silicosis by intratracheal silica injection for 28 days and determined hydroxyproline levels to evaluate collagen metabolism in lung homogenates. Oxidative stress status was evaluated by detecting catalase and glutathione peroxidase activities.
文摘To investigate the mechanisms of microwave induced pacemaker cell injuries, Wistar rats and the primary pacemaker cells of newborn Wistar rats were exposed to microwave at average power density of 50 mW/cm2. Slower spontaneous beating rate, intercellular Ca2+ aggregation and cell membrane perforation were detected immediately after the exposure. Moreover, hyperpolarizationactivated cyclic nucleotide-gated cation channel 4 (HCN4) was down-regulated immediately after the exposure and up-regulated at 12 h after the exposure. In the sinoatrial node (SAN) of the rats,
文摘The hedgehog (Hh) signaling pathway plays an essential role in the embryonic development and homeostasis of diverse adult tissues, and its deregulation has been implicated in the tumorigenesis and metastasis of various malignancies including breast cancer. Aberrant activation of the Hh pathway includes the following mechanisms: (I) Hh ligand-independent mechanism - Loss of function mutations in the Hh receptor Patched 1 (PTCH1) or gain of function mutations in the Smoothened (SMO) lead to constitutive activation of this pathway; (II) Autocrine signaling- Ith ligand produced by tumor cells stimulates the Hh signaling in tumor cells; (III) Paracrine signaling - tumor cell produced-Hh ligand activates stromal and endothelial cells that produce growth factors in microenvironment for supporting tumor growth and survival; and (IV) Reverse paracrine signaling - Hh ligand produced by stromal cells support tumor growth and survival. Upon the pathway activation, the Gli transcription factors, effectors of the Hh signaling, activate or inhibit transcription by binding to their responsive genes and interacting with the transcriptional complex. The Gli transcription factor family includes Glil, Gli2, and Gli3 (1). Glil is a transcriptional activator whose expression has been recognized as an activation state of the Hh signaling pathway, Gli2 is either an activator or repressor, and Gli3 is a strong repressor of transcriptional activities. To date, a ligand-dependent autocrine model of activating the Hh signaling has been described in breast cancer, and both an autocrine and paracrine mechanisms in colorectal cancer, pancreatic cancer and prostate cancer (2,3). Notably, a ligand-independent mechanism (mutationsin PTCHI and SMO) of the signaling has been well demonstrated in basal cell carcinoma and medulloblastoma (4,5).
文摘Background The cellular repressor of ElA-stimulated genes(CREG) is a secreted glycoprotein that inhibits cell proliferation and/or enhances differentiation.CREG is widely expressed in adult tissues such as the brain,heart, lungs,liver,intestines and kidneys in mice.We investigated the level of CREG expression during mouse embryogenesis and its distribution at 18.5 days post coitus(dpc).Methods Immunohistochemical staining with diaminobenzidine,western blotting and reverse transcription-polymerase chain reaction were used.Results CREG expression was rst detected in mouse embryos at 4.5 dpc.It was expressed at almost all stages up to 18.5 dpc.The level of CREG was found to increase gradually and was highest at 18.5 dpc.Western blotting showed that the CREG protein was expressed at higher levels in the brain,heart,intestines and kidneys than in the lungs and liver at 18.5 dpc.In 9.5 dpc embryos,CREG was expressed only in the endothelial cells of blood vessels,after the vascular lumen had formed.With advanced differentiation, vascular smooth muscle cells developed in the embryonic vascular structures;the expression of smooth muscle a-actin protein and CREG were positive and increased gradually in 10.5 dpc embryonic vessels.CREG expression in the embryonic blood vessels peaked at 15.5 dpc and was reduced slightly at 18.5 dpc.Conclusions These results indicate that CREG is expressed during mouse embryogenesis and might participate in the differentiation of these organs during embryogenesis.
基金the Scientific andTechnological DevelopmentProgram of Qingdao City, No.No.05-1-NS-73
文摘BACKGROUND: Hypoxia inducible factor-1 alpha (HIF-1 (x) and erythropoietin(EPO), possessing neuroprotective effect in the cerebral ischemia, might play an important role in the formation of cerebral ischemic tolerance (IT). OBJECTIVE:To observe the neuroprotective effect of cerebral ischemic preconditioning(IPC) of rats, and the expression and mechanism of HIF-1α and target gene erythropoietin in the brain tissue following the formation of cerebral IT. DESIGN : A randomized and controlled observation SETTING: Department of Neurology, the Affiliated Hospital of Medical College, Qingdao University MATERIALS: Totally 84 enrolled adult healthy male Wistar rats of clean grade, weighing 250 to 300 g, were provided by the Animal Experimental Department, Tongji Medical College of Huazhong University of Science and Technology. Ready-to-use SABC reagent kit and rabbit anti-rat HIF-1α monoclonal antibody were purchased from Boshide Bioengineering Co.Ltd (Wuhan); Rabbit anti-rat EPO monoclonal antibody was purchased from Santa Cruz Company (USA). METHODS: This experiment was carried out in the Department of Anatomy, Medical College, Qingdao University during March 2005 to March 2006. ① The 84 rats were divided into 3 groups by a lot: IPC group (n=40), sham-operation group (n=40) and control group (n=4). In the IPC group, middle cerebral artery was occluded for 2 hours respectively on the 1^st, 3^rd, 7^th, 14^th and 21^st days of the reperfusion following 10-minute preischemia was made using a modified middle cerebral artery second suture method from Zea-Longa. The rats were sacrificed 22 hours after reperfusion in the end of middle cerebral artery occlusion (MCAO). That was to say, after 10-minute preischemia, suture was exited to the extemal carotid artery and embedded subcutaneously. Middle cerebral artery was occluded again to form the second reperfusion at the set time point after reperfusion. Twenty-two hours later, rats were sacrificed; In the sham-operation group,the preischemia was substituted by sham-operation(only common carotid artery and crotch were exposed, and MCAO by suture was omitted), and the other procedures were the same as those in the IPC group. In the control group, rats were given sham-operation twice at an interval of one day, and they were sacrificed 24 hours after the second sham-operation. ② Brain tissue was taken from the rats in each group. Cerebral infarction area of each layer was measured with TTC staining, and total cerebral infarction volume (The total cerebral infarction area of each layerxinterspace ) was calculated. After brain tissue was stained by haematoxylin-esoin (HE), the form of nerve cells was observed under an optical microscope, and the expressions of HIF-1α(and EPO protein in the brain tissue were detected with immunohistochemical method. MAIN OUTCOME MEASURES: ①Cerebral infarction volume;②form of nerve cell; ③ the expression of HIF-1α and EPO protein in the brain tissue. RESULTS:Totally 84 rats were enrolled in the experiment. The dead rats were randomly supplied during the experiment, and finally 84 rats entered the stage of result analysis. ① Detection of cerebral infarction volume of rats in each group: Cerebral infarction volume in the IPC group was significantly smaller than that in the sham-operation group on the 1^st, 3^rd and 7^th days after reperfusion respectively [(161.2±6.9) mm^3 vs (219.9±11.2) mm^3, (134.9±9.0) mm^3 vs (218.6±13.0) mm^3, (142.9±13.7) mm^3 vs (221.3±14.2) mm^3, t=-8.924, 10.587,7.947, P〈 0.01]. ② Observation of nerve cell form of brain tissue: HE staining showed that the ischemic degree, range and cerebral edema degree of IPC group were significantly milder than those of sham-operation group. ③ The expressions of HIF-1α and EPO protein in cerebral cortex and hippocampus : The expression of HIF-1αof IPC group was significantly higher than that of sham-operation group on the 1^st, 3^rd and 7^th days after reperfusion respectively (125.93±3.79 vs 117.65±5.60, 140.63±4.64 vs 119.33±4.26, 131.15±2.74 vs 107.60±3.89, t=2.449, 6.763,9.899,P 〈 0.05-0.01). The expression of EPO of IPC group was significantly higher than that of sham-operation group on the 3^rd and 7^th days after perfusion respectively (141.68±3.29 vs 126.33±4.51, 138.88±2.59 vs 125.58±6.18,t=5.499,3.970, P〈 0.05). CONCLUSION : ①IPC can protect the never cells in rat brain and the best time to onset of cerebral IT induced by IPC is 1 to 7 days after reperfusion. ② Neuroprotective effect of cerebral IT might be related to the expression of HIF-1α and its target gene EPO.