Trastuzumab resistance is one of the causes of poor prognosis in patients with human epidermal growth factor receptor 2(HER2)-positive(HER2+)breast cancer(BC).The truncated isoform of dopamine-and cAMPregulated phosph...Trastuzumab resistance is one of the causes of poor prognosis in patients with human epidermal growth factor receptor 2(HER2)-positive(HER2+)breast cancer(BC).The truncated isoform of dopamine-and cAMPregulated phosphoprotein(t-DARPP)has been reported to be involved in trastuzumab therapy resistance and promoting tumor progression.To evaluate the t-DARPP expression in BC,paired tumors and surrounding normal tissues were analyzed by real-time polymerase chain reaction and confirmed higher DARPP-32 kDa family mRNA expression in HER2+BC tumor tissues.We established 2 patient-derived xenografts(PDX)mice models to test the efficacy of trastuzumab,named model 1(non-responder)and model 2(responder).t-DARPP and p95-HER2 protein-protein interactions were detected in PDX tumor tissue from non-responders using Förster resonance energy transfer assays.Instead,there is no response from the responder.Furthermore,mechanistic studies using transwell and western blot assays demonstrated that t-DARPP could upregulate epithelial-mesenchymal transition signaling proteins,enhance p95-HER2 expression and promote cell migration.We found that quercetin effectively reduced t-DARPP expression in HER2+BC cells.In t-DARPP ShRNA-suppressed cells,quercetin synergistically enhanced trastuzumab-induced apoptotic cell death and G2/M phase arrest.In conclusion,the combination of quercetin and trastuzumab treatment by targeting t-DARPP in HER2+BC patients has the potential as a biomarker for mitigating drug resistance.展开更多
Type I interferon(IFN-I)provides an important first line to protect avian species against pathogens invasion.IFN regulatory factor 7(IRF7)has been identified as the most important regulator for both DNA and RNA virus-...Type I interferon(IFN-I)provides an important first line to protect avian species against pathogens invasion.IFN regulatory factor 7(IRF7)has been identified as the most important regulator for both DNA and RNA virus-induced IFN-I production in chickens.Although four splicing variants of IRF7 have been identified in mammals,it is still unclear whether alternative splicing patterns and the function of IRF7 isoform(s)exist in chickens.In this study,we reported a novel short transcript isoform of chicken IRF7(chIRF7),termed chIRF7-iso,which contained an intact N-terminal DNA-binding domain(DBD)and 14 amino acids different from chIRF7 in the C-terminal.Overexpression of chIRF7 in chicken leghorn male hepatocellular(LMH)cells activated the IFN-βpromoter and significantly inhibited Newcastle disease virus(NDV)and fowl adenovirus serotype 4(FAdV-4)replication.Conversely,overexpression of chIRF7-iso blocked the IFN-βpromoter activity and was favorable for NDV and FAdV-4 replication in vitro.Collectively,our results confirm that a novel chIRF7 isoform-mediated negative regulates IFN-βproduction,which will contribute to understanding the role of chIRF7 in innate antiviral response in chicken.展开更多
Leaf acid phosphatase (APase) activities of 274 soybean genotypes were surveyed under field conditions with two levels of P supplies, and a nutrient solution culture experiment with eight selected genotypes was subseq...Leaf acid phosphatase (APase) activities of 274 soybean genotypes were surveyed under field conditions with two levels of P supplies, and a nutrient solution culture experiment with eight selected genotypes was subsequently conducted under greenhouse conditions to further characterize APase activity and its isoform expression induced by P starvation. Results from the field experiment showed that there was a great genotypic variation for leaf APase activity among the tested soybean genotypes from different origins, and APase activity in many of the tested genotypes (about 60%) was generally increased in the treatment without P fertilizer addition. Results from the nutrient solution culture experiment showed that APase activity in all the eight tested genotypes was generally enhanced by P starvation. Six isoforms of APases were detected in isoelectric focusing gels with samples from both young and old leaves. The activity of all the six isoforms was increased by P starvation, but no new APase isoform was induced. Our results suggest that leaf APase activity could serve as an enzymatic indicator of P starvation for soybean; the increase in leaf APase activity under low P stress was mainly caused by the increase in the activity of existing isoforms but not by the induction of new isoforms.展开更多
Objective: To investigate the role of PKC isoforms in the regulation of LPS-triggered tumoricidal activity in macrophages and further elucidate its signal mechanisms. Methods: Two macrophage cell lines (P388D1 and RAW...Objective: To investigate the role of PKC isoforms in the regulation of LPS-triggered tumoricidal activity in macrophages and further elucidate its signal mechanisms. Methods: Two macrophage cell lines (P388D1 and RAW264.7) were stimulated by LPS alone, or with long-term of PMA pretreatment. Then cytotoxicities to P815 cells (by MTT assay) and IL-1, TNF- (by ELISA) and nitric oxide (NO) production (by Griess reagent) in supernatants were measured. Western blot for PKC isoforms after long-term PMA pretreatment was analyzed. Results: RAW264.7 cells were stimulated with LPS to kill target tumor cells P815, whereas P388D1 cells failed to develop such an ability. Down-regulation of PKC isoforms by chronic treatment with PMA significantly inhibited the LPS-induced cytotoxicity in RAW264.7 cells. In unstimulated state, Western blotting with rabbit antiserum specific for the PKC, 1, 2, or showed all 5 isoforms were detected in P388D1 cells, while only PKC, PKC1 and PKC were detected in RAW264.7 cells. Exposure of the cells to long-term of PMA treatment significantly down-regulated the expression of PKC, PKC1 and PKC in RAW264.7 cells. But in P388D1 cells, although PKC, PKC and PKC were down-regulated, the expression of PKC1 and PKC2 could not be regulated. Comparing with LPS-induced IL-1, TNF- and NO production by the two macrophage cell lines, P388D1 failed to produce NO. In RAW264.7 cells, LPS-induced NO production and antitumor activity was attenuated by the addition of L-NAME, an iNOS inhibitor. Conclusion: The results indicated a critical role of PKC in LPS-induced antitumor activity and this cytotoxicity is mainly due to PKC- mediated NO production by RAW264.7 cells, but not a direct cytotoxic activity.展开更多
[Objective] This study aimed to analyze the effects of temperature on the expression of AGPase isoform genes in rice endosperm during milk stage. [Method] Different temperature treatments (33 and 25 ℃ of daily mean ...[Objective] This study aimed to analyze the effects of temperature on the expression of AGPase isoform genes in rice endosperm during milk stage. [Method] Different temperature treatments (33 and 25 ℃ of daily mean temperature for high and normal temperature treatments, respectively) and the real-time fluorescence quantitative PCR ( FQPCR) were used to analyze the expression patterns of seven isoforms (AGPS1, AGPS2a, AGPS2b, AGPL1, AGPL2, AGPL3 and AGPL4) of ADPglucose pyrophosphorylase (AGPase) which was the key enzyme in starch synthesis and metabolism in rice endosperm of two rice varieties Teqing and Thai Fragrant Rice. [Result] The AGPase isoforms AGPS2b, AGPL2 and AGPL3 had much higher expression than the other four isoforms, thus they were thought to be the main expression patterns of AGPase in rice endosperm. The relative expressions of AGPL2 was the highest among all the isoforms. The relative expressions of AGPS2b, AGPL2 and AGPL3 were higher in the normal temperature treatment than in the high temperature treatment in both rice varieties. The relative expression of the three enzyme genes in milk stages in Teqing was higher than those in Thai Fragrant Rice under different temperature treatments. [Conclusion] This study provides a theoretical basis for further use of molecular biology techniques to cultivate stable high-quality rice varieties.展开更多
Cardiac arrest can lead to severe neurological impairment as a result of inflammation,mitochondrial dysfunction,and post-cardiopulmonary resuscitation neurological damage.Hypoxic preconditioning has been shown to impr...Cardiac arrest can lead to severe neurological impairment as a result of inflammation,mitochondrial dysfunction,and post-cardiopulmonary resuscitation neurological damage.Hypoxic preconditioning has been shown to improve migration and survival of bone marrow–derived mesenchymal stem cells and reduce pyroptosis after cardiac arrest,but the specific mechanisms by which hypoxia-preconditioned bone marrow–derived mesenchymal stem cells protect against brain injury after cardiac arrest are unknown.To this end,we established an in vitro co-culture model of bone marrow–derived mesenchymal stem cells and oxygen–glucose deprived primary neurons and found that hypoxic preconditioning enhanced the protective effect of bone marrow stromal stem cells against neuronal pyroptosis,possibly through inhibition of the MAPK and nuclear factor κB pathways.Subsequently,we transplanted hypoxia-preconditioned bone marrow–derived mesenchymal stem cells into the lateral ventricle after the return of spontaneous circulation in an 8-minute cardiac arrest rat model induced by asphyxia.The results showed that hypoxia-preconditioned bone marrow–derived mesenchymal stem cells significantly reduced cardiac arrest–induced neuronal pyroptosis,oxidative stress,and mitochondrial damage,whereas knockdown of the liver isoform of phosphofructokinase in bone marrow–derived mesenchymal stem cells inhibited these effects.To conclude,hypoxia-preconditioned bone marrow–derived mesenchymal stem cells offer a promising therapeutic approach for neuronal injury following cardiac arrest,and their beneficial effects are potentially associated with increased expression of the liver isoform of phosphofructokinase following hypoxic preconditioning.展开更多
Aim: To investigate the androgen receptor (AR) isoforms and its variability of expression in human and rat prostatictissues. Methods: Human benign prostatic hyperplasia (BPH) and prostatic cancer tissues were obtained...Aim: To investigate the androgen receptor (AR) isoforms and its variability of expression in human and rat prostatictissues. Methods: Human benign prostatic hyperplasia (BPH) and prostatic cancer tissues were obtained from pa-tients undergoing prostatectomy, and rat ventral prostate was incised 3 days after castration. Forty-one AR-positive BPHspecimens, 3 prostatic cancer specimens, and 6 rat prostates were used. After processing at 4℃, the tissues were ex-amined by means of high resolution isoelectric focusing (IEF) technique to determine their AR isoforms. Results:From the prostatic specimens, 3 types of AR isoforms were detected with pI values at 6.5, 6.0, and 5.3. In humanBPH tissues, 15/41 (36.6%) specimens showed all the three types of isoforms, while 19/41 (46.3%) showed 2 iso-fora at various combinations and 7/41 (17.1%), 1 isoform. For the 3 prostatic cancer specimens, one showed 3 iso-forms, one, 2 isoforms, and the other failed to show any isoform. All rat prostatic tissues showed 2 isoforms at differ-ent combinations. Binding of ~3H-dihydrotestosterone (DHT) to the isoforms was inhibited by the addition of 100-foldexcess of DHT or testosterone, but not progesterone, oestradiol or diethylstilboestrol. Conclusion: AR isoforms aredifferent in different patients. Although their genesis is not clear, the therapeutic implication of the present observationappears to be interesting, that may help clarifying the individual differences in the response to hormonal therapy.(Asian J Androl 2000 Dec; 2: 307-310)展开更多
Protein arginine methyltransferases(PRMTs) catalyze the methylation of a variety of protein substrates, many of which have been linked to the development, progression and aggressiveness of different types of cancer. M...Protein arginine methyltransferases(PRMTs) catalyze the methylation of a variety of protein substrates, many of which have been linked to the development, progression and aggressiveness of different types of cancer. Moreover, aberrant expression of PRMTs has been observed in several cancer types. While the link between PRMTs and cancer is a relatively new area of interest, the functional implications documented thus far warrant further investigations into its therapeutic potential. However, the expression of these enzymes and the regulation of their activity in cancer are still significantly understudied. Currently there are nine main members of the PRMT family. Further, the existence of alternatively spliced isoforms for several of these family members provides an additional layer of complexity. Specifically, PRMT1, PRMT2, CARM1 and PRMT7 have been shown to have alternative isoforms and others may be currently unrealized. Our knowledge with respect to the relative expression and the specific functions of these isoforms is largely lacking and needs attention. Here we present a review of the current knowledge of theknown alternative PRMT isoforms and provide a rationale for how they may impact on cancer and represent potentially useful targets for the development of novel therapeutic strategies.展开更多
In this study, the gene encoding hemocyanin subunit L, Lv Hc L, was cloned from Litopenaeus vannamei and the genomic organization was characterized. This gene was diverse with many SNPs and also had at least four isof...In this study, the gene encoding hemocyanin subunit L, Lv Hc L, was cloned from Litopenaeus vannamei and the genomic organization was characterized. This gene was diverse with many SNPs and also had at least four isoforms, while one of them(Lv Hc L4) only had two exons and the exon2 was missed. Transcription analysis showed that these isoforms of Lv Hc L were up-regulated after WSSV challenge in WSSV-resistant shrimp, while the transcriptions were decreased constantly in WSSV-susceptible shrimp. It is suggested that the hemocyanin had rich polymorphism and was involved in the antiviral response. These results could extend our previous findings and provide insights into the immune feature of hemocyanin, which would be helpful for further studies aimed at antiviral mechanism in invertebrate.展开更多
AIM:To investigate three isoforms of survivin in colorectal adenocarcinomas.METHODS:We used the LightCycler Technology(Roche),along with a common forward primer and reverse primers specific for the splice variants and...AIM:To investigate three isoforms of survivin in colorectal adenocarcinomas.METHODS:We used the LightCycler Technology(Roche),along with a common forward primer and reverse primers specific for the splice variants and two common hybridization probes labeled with fluorescein and LightCycler-Red fluorophore(LC-Red 640).Real time quantitative polymerase chain reaction(PCR) was performed on cDNAs from 52 tumor specimens from colorectal cancer patients and 10 unrelated normal colorectal tissues.In the patients group,carcinoembryonic antigen(CEA) and CA19-9 tumor markers were also measured immunochemically.RESULTS:Wild type survivin mRNA isoform was expressed in 48%of the 52 tumor samples,survivin-2b in 38%and survivin-ΔΕx3 in 29%,while no expression was found in normal tissues.The mRNA expression of wild type survivin presented a significant correlation with the expression of the ratio of survivin-2b,survivin-ΔΕx3,survivin-2b/wild type survivin and survivin-ΔΕx3/wild type survivin(P<0.001).The mRNA expression of wildsurvivin and survivin-ΔΕx3 was related with tumor size and invasion(P=0.006 and P<0.005,respectively).A significant difference was found between survivin-2b and morphologic cancer type.Also,the ratio of survivin-ΔEx3/ wild-survivin was significantly associated with prognosis.No association was observed between the three isoforms and grade,metastasis,Dukes stage and gender.The three isoforms were not correlated with CEA and CA19-9.CONCLUSION:Survivin isoforms may play a role in cell apoptosis and their quantification could provide information about clinical management of patients suffering from colorectal cancer.展开更多
Nitrogen is a key element to control the growth and yield of crops. Fertilizer urea nitrogen (N) 60,45, and 30 kg/hm2 was applied at three different stages, midtillering, panicle initiation, and flowering, of the grow...Nitrogen is a key element to control the growth and yield of crops. Fertilizer urea nitrogen (N) 60,45, and 30 kg/hm2 was applied at three different stages, midtillering, panicle initiation, and flowering, of the growth and development of rice plants, respectively. At both midtillering and panicle initiation, the total activity of glutamine synthetase (GS) in rice roots and leaves was incrased remarkably as a result of a large amount of ammonia absorbed by roots. Native-PAGE and activity staining showed that the increase of total activity in rice roots and leaves was due to the synthesis of GSrb in roots and GS2 in leaves and that the activity of GSra in roots and GS1 in leaves remained constant. The results showed that the assimilation of external nitrogen was carried out by GSrb but not GSra in rice roots and that the activitry of GS2 was induced also by the external nitrogen, and that GSrb played main role in meeting the needs of the rapid tillering for nitrogen. At flowering, the activity of GS in rice roots and leaves did not change almost after topdressing. These rssults suggest that the change of GS activity in rice roots may use as a measure of the utilization efficiency of the fertilizer.展开更多
AIM To investigate the role of Δ133p53 isoform in nuclear factor-κB(NF-κB) inhibitor pyrrolidine dithiocarbamate(PDTC)-mediated growth inhibition of MKN45 gastric cancer cells.METHODS The growth rate of MKN45 cells...AIM To investigate the role of Δ133p53 isoform in nuclear factor-κB(NF-κB) inhibitor pyrrolidine dithiocarbamate(PDTC)-mediated growth inhibition of MKN45 gastric cancer cells.METHODS The growth rate of MKN45 cells after treatment with different concentrations of only PDTC or PTDC in combination with cisplatin was detected by the CCK-8 assay. m RNA expression levels of Δ133p53, p53β, and the NF-κB p65 subunit and p65 protein levels were detected by reverse transcription-polymerase chain reaction(RT-PCR) and immunofluorescence, respectively. Growth of MKN45 cells was significantly inhibited by PDTC alone in a dose-dependent manner(P < 0.01). Moreover, the inhibitory effect of cisplatin was remarkably enhanced in a dose-dependent manner by co-treatment with PDTC(P < 0.01).RESULTS RT-PCR analysis revealed that m RNA expression of p65 was curbed significantly in a dose-dependent manner by treatment with only PDTC(P < 0.01), and this suppressive effect was further enhanced when co-treated with cisplatin(P < 0.01). With respect to the other p53 isoforms, m RNA level of Δ133p53 was significantly reduced in a dose-dependent manner by treatment with only PDTC or PTDC in combination with cisplatin(P < 0.01), whereas p53β m RNA expression was not altered by PDTC treatment(P > 0.05). A similar tendency of change in p65 protein expression, as observed for the corresponding m RNA, was detected by immunofluorescence analysis(P < 0.01). Pearson correlation analysis demonstrated that Δ133p53 and p65 m RNA expression levels were positively related, while no significant relationship was observed between those of p65 and p53β(r = 0.076, P > 0.01).CONCLUSIONΔ133p53 isoform(not p53β) is required in PDTCinduced inhibition of MKN45 gastric cancer cells, indicating that disturbance in the cross-talk between p53 and NF-κB pathways is a promising target in pharmaceutical research for the development of treatment strategies for gastric cancer.展开更多
AIM:To analyze the risk of coronary heart disease in patients with type 2 diabetes mellitus(T2DM)receiving standard medical treatment.METHODS:We performed a retrospective chart analysis of 269 middle-aged patients(age...AIM:To analyze the risk of coronary heart disease in patients with type 2 diabetes mellitus(T2DM)receiving standard medical treatment.METHODS:We performed a retrospective chart analysis of 269 middle-aged patients(age 45-64 years,mean age,53.9±5.5 years)with T2DM and without atherosclerotic cardiovascular events who underwent typing to determine their apolipoprotein E(apoE)isoforms.The apoE isoforms were determined using isoelectric focusing,followed by immunoblotting.We retrospectively evaluated the charts of the 269 patients,recorded between their first visit to the hospital(the study's start point,between 1987 and 1992)and the occurrence of an atherosclerotic cardiovascular event(the study's endpoint)or January 2004,whichever came first.The age-adjusted mean values and the prevalences of covariates were calculated to compare the laboratory data among the apoE phenotypes.To investigate the association of risk factors with the incidence of coronary heart disease during the follow-up period,monovariate and multivariate Cox regression models were used.RESULTS:At enrollment,the mean serum low density lipoprotein(LDL)cholesterol levels were lowest(2.92± 0.89 mmol/L)among the subjects with apoE2(apoE2/2 or apoE2/3)and highest(3.52±0.77 mmol/L)among the subjects with apoE4(apoE3/4 or apoE4/4).No significant differences in mean age or the percentage of smokers were observed among the three groups.Furthermore,no significant differences were observed in the systolic and diastolic blood pressures,body mass index,HbA1c level or serum triglyceride levels among the three groups.There were 47 cases of coronary heart disease over 3285 person-years of follow-up.An age-adjusted multivariate Cox proportional model identified diabetic retinopathy(hazard ratio,2.38,95% CI:1.28-4.43,P=0.006),a high systolic blood pressure(hazard ratio,1.04,95%CI:1.02-1.06,P<0.001) and high HbA1c values(hazard ratio,1.19,95%CI:1.02-1.38,P=0.0029),but not the LDL cholesterol value at enrollment(hazard ratio,1.01,95%CI:0.97-1.05,P=0.77)nor the specific apoE isoform,as significant predictors of coronary heart disease.CONCLUSION:Under standard medical treatment of diabetes,including the control of LDL cholesterol levels,the apoE4 isoform was not associated with coronary heart disease among T2DM patients.展开更多
基金The National Science and Technology Council of Taiwan funded this study.
文摘Trastuzumab resistance is one of the causes of poor prognosis in patients with human epidermal growth factor receptor 2(HER2)-positive(HER2+)breast cancer(BC).The truncated isoform of dopamine-and cAMPregulated phosphoprotein(t-DARPP)has been reported to be involved in trastuzumab therapy resistance and promoting tumor progression.To evaluate the t-DARPP expression in BC,paired tumors and surrounding normal tissues were analyzed by real-time polymerase chain reaction and confirmed higher DARPP-32 kDa family mRNA expression in HER2+BC tumor tissues.We established 2 patient-derived xenografts(PDX)mice models to test the efficacy of trastuzumab,named model 1(non-responder)and model 2(responder).t-DARPP and p95-HER2 protein-protein interactions were detected in PDX tumor tissue from non-responders using Förster resonance energy transfer assays.Instead,there is no response from the responder.Furthermore,mechanistic studies using transwell and western blot assays demonstrated that t-DARPP could upregulate epithelial-mesenchymal transition signaling proteins,enhance p95-HER2 expression and promote cell migration.We found that quercetin effectively reduced t-DARPP expression in HER2+BC cells.In t-DARPP ShRNA-suppressed cells,quercetin synergistically enhanced trastuzumab-induced apoptotic cell death and G2/M phase arrest.In conclusion,the combination of quercetin and trastuzumab treatment by targeting t-DARPP in HER2+BC patients has the potential as a biomarker for mitigating drug resistance.
基金supported by the grants from the National Natural Science Foundation of China(32002259)the Natural Science Foundation of Henan Province,China(202300410198).
文摘Type I interferon(IFN-I)provides an important first line to protect avian species against pathogens invasion.IFN regulatory factor 7(IRF7)has been identified as the most important regulator for both DNA and RNA virus-induced IFN-I production in chickens.Although four splicing variants of IRF7 have been identified in mammals,it is still unclear whether alternative splicing patterns and the function of IRF7 isoform(s)exist in chickens.In this study,we reported a novel short transcript isoform of chicken IRF7(chIRF7),termed chIRF7-iso,which contained an intact N-terminal DNA-binding domain(DBD)and 14 amino acids different from chIRF7 in the C-terminal.Overexpression of chIRF7 in chicken leghorn male hepatocellular(LMH)cells activated the IFN-βpromoter and significantly inhibited Newcastle disease virus(NDV)and fowl adenovirus serotype 4(FAdV-4)replication.Conversely,overexpression of chIRF7-iso blocked the IFN-βpromoter activity and was favorable for NDV and FAdV-4 replication in vitro.Collectively,our results confirm that a novel chIRF7 isoform-mediated negative regulates IFN-βproduction,which will contribute to understanding the role of chIRF7 in innate antiviral response in chicken.
文摘Leaf acid phosphatase (APase) activities of 274 soybean genotypes were surveyed under field conditions with two levels of P supplies, and a nutrient solution culture experiment with eight selected genotypes was subsequently conducted under greenhouse conditions to further characterize APase activity and its isoform expression induced by P starvation. Results from the field experiment showed that there was a great genotypic variation for leaf APase activity among the tested soybean genotypes from different origins, and APase activity in many of the tested genotypes (about 60%) was generally increased in the treatment without P fertilizer addition. Results from the nutrient solution culture experiment showed that APase activity in all the eight tested genotypes was generally enhanced by P starvation. Six isoforms of APases were detected in isoelectric focusing gels with samples from both young and old leaves. The activity of all the six isoforms was increased by P starvation, but no new APase isoform was induced. Our results suggest that leaf APase activity could serve as an enzymatic indicator of P starvation for soybean; the increase in leaf APase activity under low P stress was mainly caused by the increase in the activity of existing isoforms but not by the induction of new isoforms.
文摘Objective: To investigate the role of PKC isoforms in the regulation of LPS-triggered tumoricidal activity in macrophages and further elucidate its signal mechanisms. Methods: Two macrophage cell lines (P388D1 and RAW264.7) were stimulated by LPS alone, or with long-term of PMA pretreatment. Then cytotoxicities to P815 cells (by MTT assay) and IL-1, TNF- (by ELISA) and nitric oxide (NO) production (by Griess reagent) in supernatants were measured. Western blot for PKC isoforms after long-term PMA pretreatment was analyzed. Results: RAW264.7 cells were stimulated with LPS to kill target tumor cells P815, whereas P388D1 cells failed to develop such an ability. Down-regulation of PKC isoforms by chronic treatment with PMA significantly inhibited the LPS-induced cytotoxicity in RAW264.7 cells. In unstimulated state, Western blotting with rabbit antiserum specific for the PKC, 1, 2, or showed all 5 isoforms were detected in P388D1 cells, while only PKC, PKC1 and PKC were detected in RAW264.7 cells. Exposure of the cells to long-term of PMA treatment significantly down-regulated the expression of PKC, PKC1 and PKC in RAW264.7 cells. But in P388D1 cells, although PKC, PKC and PKC were down-regulated, the expression of PKC1 and PKC2 could not be regulated. Comparing with LPS-induced IL-1, TNF- and NO production by the two macrophage cell lines, P388D1 failed to produce NO. In RAW264.7 cells, LPS-induced NO production and antitumor activity was attenuated by the addition of L-NAME, an iNOS inhibitor. Conclusion: The results indicated a critical role of PKC in LPS-induced antitumor activity and this cytotoxicity is mainly due to PKC- mediated NO production by RAW264.7 cells, but not a direct cytotoxic activity.
基金Supported by Special Project for Breeding and Cultivation of GMO Varieties of Ministry of Agriculture (2011ZX08001-001, 2011ZX08001-004)Major Science and Technology Program of Hunan, China (2011FJ1002-2)+1 种基金Natural Science Foundation of Hunan, China (09JJ3046 )Science and Technology Innovation Program of Hunan Academy of Agricultural Sciences (2009hnnkycx17)~~
文摘[Objective] This study aimed to analyze the effects of temperature on the expression of AGPase isoform genes in rice endosperm during milk stage. [Method] Different temperature treatments (33 and 25 ℃ of daily mean temperature for high and normal temperature treatments, respectively) and the real-time fluorescence quantitative PCR ( FQPCR) were used to analyze the expression patterns of seven isoforms (AGPS1, AGPS2a, AGPS2b, AGPL1, AGPL2, AGPL3 and AGPL4) of ADPglucose pyrophosphorylase (AGPase) which was the key enzyme in starch synthesis and metabolism in rice endosperm of two rice varieties Teqing and Thai Fragrant Rice. [Result] The AGPase isoforms AGPS2b, AGPL2 and AGPL3 had much higher expression than the other four isoforms, thus they were thought to be the main expression patterns of AGPase in rice endosperm. The relative expressions of AGPL2 was the highest among all the isoforms. The relative expressions of AGPS2b, AGPL2 and AGPL3 were higher in the normal temperature treatment than in the high temperature treatment in both rice varieties. The relative expression of the three enzyme genes in milk stages in Teqing was higher than those in Thai Fragrant Rice under different temperature treatments. [Conclusion] This study provides a theoretical basis for further use of molecular biology techniques to cultivate stable high-quality rice varieties.
基金supported by the Natural Science Fund of Fujian Province,No.2020J011058(to JK)the Project of Fujian Provincial Hospital for High-level Hospital Construction,No.2020HSJJ12(to JK)+1 种基金the Fujian Provincial Finance Department Special Fund,No.(2021)848(to FC)the Fujian Provincial Major Scientific and Technological Special Projects on Health,No.2022ZD01008(to FC).
文摘Cardiac arrest can lead to severe neurological impairment as a result of inflammation,mitochondrial dysfunction,and post-cardiopulmonary resuscitation neurological damage.Hypoxic preconditioning has been shown to improve migration and survival of bone marrow–derived mesenchymal stem cells and reduce pyroptosis after cardiac arrest,but the specific mechanisms by which hypoxia-preconditioned bone marrow–derived mesenchymal stem cells protect against brain injury after cardiac arrest are unknown.To this end,we established an in vitro co-culture model of bone marrow–derived mesenchymal stem cells and oxygen–glucose deprived primary neurons and found that hypoxic preconditioning enhanced the protective effect of bone marrow stromal stem cells against neuronal pyroptosis,possibly through inhibition of the MAPK and nuclear factor κB pathways.Subsequently,we transplanted hypoxia-preconditioned bone marrow–derived mesenchymal stem cells into the lateral ventricle after the return of spontaneous circulation in an 8-minute cardiac arrest rat model induced by asphyxia.The results showed that hypoxia-preconditioned bone marrow–derived mesenchymal stem cells significantly reduced cardiac arrest–induced neuronal pyroptosis,oxidative stress,and mitochondrial damage,whereas knockdown of the liver isoform of phosphofructokinase in bone marrow–derived mesenchymal stem cells inhibited these effects.To conclude,hypoxia-preconditioned bone marrow–derived mesenchymal stem cells offer a promising therapeutic approach for neuronal injury following cardiac arrest,and their beneficial effects are potentially associated with increased expression of the liver isoform of phosphofructokinase following hypoxic preconditioning.
文摘Aim: To investigate the androgen receptor (AR) isoforms and its variability of expression in human and rat prostatictissues. Methods: Human benign prostatic hyperplasia (BPH) and prostatic cancer tissues were obtained from pa-tients undergoing prostatectomy, and rat ventral prostate was incised 3 days after castration. Forty-one AR-positive BPHspecimens, 3 prostatic cancer specimens, and 6 rat prostates were used. After processing at 4℃, the tissues were ex-amined by means of high resolution isoelectric focusing (IEF) technique to determine their AR isoforms. Results:From the prostatic specimens, 3 types of AR isoforms were detected with pI values at 6.5, 6.0, and 5.3. In humanBPH tissues, 15/41 (36.6%) specimens showed all the three types of isoforms, while 19/41 (46.3%) showed 2 iso-fora at various combinations and 7/41 (17.1%), 1 isoform. For the 3 prostatic cancer specimens, one showed 3 iso-forms, one, 2 isoforms, and the other failed to show any isoform. All rat prostatic tissues showed 2 isoforms at differ-ent combinations. Binding of ~3H-dihydrotestosterone (DHT) to the isoforms was inhibited by the addition of 100-foldexcess of DHT or testosterone, but not progesterone, oestradiol or diethylstilboestrol. Conclusion: AR isoforms aredifferent in different patients. Although their genesis is not clear, the therapeutic implication of the present observationappears to be interesting, that may help clarifying the individual differences in the response to hormonal therapy.(Asian J Androl 2000 Dec; 2: 307-310)
基金Supported by Cancer projects in the C télab are funded through the Cancer Research Society,Canadian Research Institutes of Health Research and Canadian Breast Cancer Foundation
文摘Protein arginine methyltransferases(PRMTs) catalyze the methylation of a variety of protein substrates, many of which have been linked to the development, progression and aggressiveness of different types of cancer. Moreover, aberrant expression of PRMTs has been observed in several cancer types. While the link between PRMTs and cancer is a relatively new area of interest, the functional implications documented thus far warrant further investigations into its therapeutic potential. However, the expression of these enzymes and the regulation of their activity in cancer are still significantly understudied. Currently there are nine main members of the PRMT family. Further, the existence of alternatively spliced isoforms for several of these family members provides an additional layer of complexity. Specifically, PRMT1, PRMT2, CARM1 and PRMT7 have been shown to have alternative isoforms and others may be currently unrealized. Our knowledge with respect to the relative expression and the specific functions of these isoforms is largely lacking and needs attention. Here we present a review of the current knowledge of theknown alternative PRMT isoforms and provide a rationale for how they may impact on cancer and represent potentially useful targets for the development of novel therapeutic strategies.
基金The National High Technology Research and Development Program(863 Program)of China under contract No.2012AA092205the Major State Basic Research Development Program(973 Program)of China under contract No.2012CB114403+2 种基金the China Agriculture Research System-47the National Natural Science Foundation of China under contract No.31472297 and 31302209the Scientific Research Foundation of Third Institute of Oceanography,SOA under contract No.2011018
文摘In this study, the gene encoding hemocyanin subunit L, Lv Hc L, was cloned from Litopenaeus vannamei and the genomic organization was characterized. This gene was diverse with many SNPs and also had at least four isoforms, while one of them(Lv Hc L4) only had two exons and the exon2 was missed. Transcription analysis showed that these isoforms of Lv Hc L were up-regulated after WSSV challenge in WSSV-resistant shrimp, while the transcriptions were decreased constantly in WSSV-susceptible shrimp. It is suggested that the hemocyanin had rich polymorphism and was involved in the antiviral response. These results could extend our previous findings and provide insights into the immune feature of hemocyanin, which would be helpful for further studies aimed at antiviral mechanism in invertebrate.
文摘AIM:To investigate three isoforms of survivin in colorectal adenocarcinomas.METHODS:We used the LightCycler Technology(Roche),along with a common forward primer and reverse primers specific for the splice variants and two common hybridization probes labeled with fluorescein and LightCycler-Red fluorophore(LC-Red 640).Real time quantitative polymerase chain reaction(PCR) was performed on cDNAs from 52 tumor specimens from colorectal cancer patients and 10 unrelated normal colorectal tissues.In the patients group,carcinoembryonic antigen(CEA) and CA19-9 tumor markers were also measured immunochemically.RESULTS:Wild type survivin mRNA isoform was expressed in 48%of the 52 tumor samples,survivin-2b in 38%and survivin-ΔΕx3 in 29%,while no expression was found in normal tissues.The mRNA expression of wild type survivin presented a significant correlation with the expression of the ratio of survivin-2b,survivin-ΔΕx3,survivin-2b/wild type survivin and survivin-ΔΕx3/wild type survivin(P<0.001).The mRNA expression of wildsurvivin and survivin-ΔΕx3 was related with tumor size and invasion(P=0.006 and P<0.005,respectively).A significant difference was found between survivin-2b and morphologic cancer type.Also,the ratio of survivin-ΔEx3/ wild-survivin was significantly associated with prognosis.No association was observed between the three isoforms and grade,metastasis,Dukes stage and gender.The three isoforms were not correlated with CEA and CA19-9.CONCLUSION:Survivin isoforms may play a role in cell apoptosis and their quantification could provide information about clinical management of patients suffering from colorectal cancer.
文摘Nitrogen is a key element to control the growth and yield of crops. Fertilizer urea nitrogen (N) 60,45, and 30 kg/hm2 was applied at three different stages, midtillering, panicle initiation, and flowering, of the growth and development of rice plants, respectively. At both midtillering and panicle initiation, the total activity of glutamine synthetase (GS) in rice roots and leaves was incrased remarkably as a result of a large amount of ammonia absorbed by roots. Native-PAGE and activity staining showed that the increase of total activity in rice roots and leaves was due to the synthesis of GSrb in roots and GS2 in leaves and that the activity of GSra in roots and GS1 in leaves remained constant. The results showed that the assimilation of external nitrogen was carried out by GSrb but not GSra in rice roots and that the activitry of GS2 was induced also by the external nitrogen, and that GSrb played main role in meeting the needs of the rapid tillering for nitrogen. At flowering, the activity of GS in rice roots and leaves did not change almost after topdressing. These rssults suggest that the change of GS activity in rice roots may use as a measure of the utilization efficiency of the fertilizer.
基金Supported by Shandong Provincial Award Foundation for Youth and Middle-aged Scientist,No.BS2010SW034
文摘AIM To investigate the role of Δ133p53 isoform in nuclear factor-κB(NF-κB) inhibitor pyrrolidine dithiocarbamate(PDTC)-mediated growth inhibition of MKN45 gastric cancer cells.METHODS The growth rate of MKN45 cells after treatment with different concentrations of only PDTC or PTDC in combination with cisplatin was detected by the CCK-8 assay. m RNA expression levels of Δ133p53, p53β, and the NF-κB p65 subunit and p65 protein levels were detected by reverse transcription-polymerase chain reaction(RT-PCR) and immunofluorescence, respectively. Growth of MKN45 cells was significantly inhibited by PDTC alone in a dose-dependent manner(P < 0.01). Moreover, the inhibitory effect of cisplatin was remarkably enhanced in a dose-dependent manner by co-treatment with PDTC(P < 0.01).RESULTS RT-PCR analysis revealed that m RNA expression of p65 was curbed significantly in a dose-dependent manner by treatment with only PDTC(P < 0.01), and this suppressive effect was further enhanced when co-treated with cisplatin(P < 0.01). With respect to the other p53 isoforms, m RNA level of Δ133p53 was significantly reduced in a dose-dependent manner by treatment with only PDTC or PTDC in combination with cisplatin(P < 0.01), whereas p53β m RNA expression was not altered by PDTC treatment(P > 0.05). A similar tendency of change in p65 protein expression, as observed for the corresponding m RNA, was detected by immunofluorescence analysis(P < 0.01). Pearson correlation analysis demonstrated that Δ133p53 and p65 m RNA expression levels were positively related, while no significant relationship was observed between those of p65 and p53β(r = 0.076, P > 0.01).CONCLUSIONΔ133p53 isoform(not p53β) is required in PDTCinduced inhibition of MKN45 gastric cancer cells, indicating that disturbance in the cross-talk between p53 and NF-κB pathways is a promising target in pharmaceutical research for the development of treatment strategies for gastric cancer.
文摘AIM:To analyze the risk of coronary heart disease in patients with type 2 diabetes mellitus(T2DM)receiving standard medical treatment.METHODS:We performed a retrospective chart analysis of 269 middle-aged patients(age 45-64 years,mean age,53.9±5.5 years)with T2DM and without atherosclerotic cardiovascular events who underwent typing to determine their apolipoprotein E(apoE)isoforms.The apoE isoforms were determined using isoelectric focusing,followed by immunoblotting.We retrospectively evaluated the charts of the 269 patients,recorded between their first visit to the hospital(the study's start point,between 1987 and 1992)and the occurrence of an atherosclerotic cardiovascular event(the study's endpoint)or January 2004,whichever came first.The age-adjusted mean values and the prevalences of covariates were calculated to compare the laboratory data among the apoE phenotypes.To investigate the association of risk factors with the incidence of coronary heart disease during the follow-up period,monovariate and multivariate Cox regression models were used.RESULTS:At enrollment,the mean serum low density lipoprotein(LDL)cholesterol levels were lowest(2.92± 0.89 mmol/L)among the subjects with apoE2(apoE2/2 or apoE2/3)and highest(3.52±0.77 mmol/L)among the subjects with apoE4(apoE3/4 or apoE4/4).No significant differences in mean age or the percentage of smokers were observed among the three groups.Furthermore,no significant differences were observed in the systolic and diastolic blood pressures,body mass index,HbA1c level or serum triglyceride levels among the three groups.There were 47 cases of coronary heart disease over 3285 person-years of follow-up.An age-adjusted multivariate Cox proportional model identified diabetic retinopathy(hazard ratio,2.38,95% CI:1.28-4.43,P=0.006),a high systolic blood pressure(hazard ratio,1.04,95%CI:1.02-1.06,P<0.001) and high HbA1c values(hazard ratio,1.19,95%CI:1.02-1.38,P=0.0029),but not the LDL cholesterol value at enrollment(hazard ratio,1.01,95%CI:0.97-1.05,P=0.77)nor the specific apoE isoform,as significant predictors of coronary heart disease.CONCLUSION:Under standard medical treatment of diabetes,including the control of LDL cholesterol levels,the apoE4 isoform was not associated with coronary heart disease among T2DM patients.