BACKGROUND: L-3-n-butylphthalide (L-NBP) can inhibit phosphorylation of tau protein and reduce the neurotoxicity of beta-amyloid peptide 1-42 (Aβ1-42). OBJECTIVE: To observe the neuroprotective effects of L-NBP...BACKGROUND: L-3-n-butylphthalide (L-NBP) can inhibit phosphorylation of tau protein and reduce the neurotoxicity of beta-amyloid peptide 1-42 (Aβ1-42). OBJECTIVE: To observe the neuroprotective effects of L-NBP on caspase-3 and nuclear factor kappa-B (NF- K B) expression in a rat model of Alzheimer's disease. DESIGN, TIME AND SETTING: A cell experiment was performed at the Central Laboratory of Provincial Hospital affiliated to Shandong University between January 2008 and August 2008. MATERIALS: L-NBP (purity 〉 98%) was provided by Shijiazhuang Pharma Group NBP Pharmaceutical Company Limited. Aβ1-42, 3-[4,5-dimethylthiazolo-2]-2,5 iphenyltetrazolium bromide (MTT), and rabbit anti-Caspase-3 polyclonal antibody were provided by Cell Signaling, USA; goat anti-choactase and rabbit anti-NF- kB antibodies were provided by Santa Cruz, USA. METHODS: Primary cultures were generated from rat basal forebrain and hippocampal neurons at 17 or 19 days of gestation. The cells were assigned into five groups: the control group, the Aβ1-42 group (2 μmol/L), the Aβ1-42 + 0.1 μmol/L L-NBP group, the Aβ1-42 + 1 μ mol/L L-NBP group, and the Aβ1-42 + 10μmol/L L-NBP group. The neurons were treated with Aβ1-42 (2 μmol/L) alone or in combination with L-NBP (0.1, 1, 10 μmol/L) for 48 hours. Cells in the control group were incubated in PBS. MAIN OUTCOME MEASURES: Morphologic changes were evaluated using inverted microscopy, viability using the M-I-I- method, and the changes in caspase-3 and NF- k B expression using Western blot. RESULTS: Induction with Aβ1-42 for 48 hours caused cell death and soma atrophy, and increased caspase-3 and NF- K B expression (P 〈 0.05). L-NBP blocked these changes in cell morphology, decreased caspase-3 and NF- k B expression (P 〈 0.05), and improved cell viability, especially at the high dose (P 〈 0.05). CONCLUSION: AI3^-42 is toxic to basal forebrain and hippocampal primary neurons; L-NBP protects against this toxicity and inhibits the induction of caspase-3 and NF- K B expression.展开更多
A pot experiment was conducted during winter growing season of 2014 at Homs Agriculture Research Center, General Commission for Scientific Researches (GCSAR), Syria. A factorial experiment arranged according to comple...A pot experiment was conducted during winter growing season of 2014 at Homs Agriculture Research Center, General Commission for Scientific Researches (GCSAR), Syria. A factorial experiment arranged according to complete randomized block design with six replications was used. A combination of four levels of saline irrigation water (tap water, 2,000, 4,000 and 6,000 ppm), with three K levels (180, 360 and 540 ppm), was used to evaluate the effects of saline irrigation water and K enrichment on some growth attributes of two sugar beet varieties (Semper and Alligator). Results showed that all studied growth attributes, i.e., leaf area (LA), leaf number (LN), total dry matter (TDM) and net assimilation rate (NAR) were decreased under salinity stress conditions compared to the control, while K enrichment significantly increased some of the studied characters such as LA, TDM and NAR, but the differences in LN were apparent according to increase in K levels. The variety Semper surpassed significantly the variety Alligator in LA, TDM and NAR. Results also indicated a significant interaction between salinity×potassium enrichment, varieties×potassium enrichment and salinity ? varieties.展开更多
Background: The carcinogenesis of hepatocellular carcinoma (HCC) is a multi-factorial, multistep and complex process. Its prognosis is poor and early detection is of the utmost importance. Transforming growth factor ...Background: The carcinogenesis of hepatocellular carcinoma (HCC) is a multi-factorial, multistep and complex process. Its prognosis is poor and early detection is of the utmost importance. Transforming growth factor β1 (TGF-β1) message RNA (mRNA) has been reported to be elevated in HCC patients using Northern blotting. However, little work has been done about the detection of TGF-β1 mRNA levels in peripheral blood of patients with HCC using the real-time polymerase chain reactions (PCR) method. Objective: To assess the prognostic value of quantitative levels of TGF-β1 mRNA in peripheral blood of patients with HCC, and to investigate the relationship between the expression of TGF-β1 mRNA in peripheral blood and many diagnostic and pathological factors. Methods: We developed an optimized Taqman real-time PCR to quantify TGF-β1 mRNA in peripheral blood of 53 patients with HCC and 44 healthy volunteers. In addition, blood was collected from patients with HCC for measuring levels of total bilirubin (TBil), prealbumin, albumin, alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma glutamyltranspeptidase (GGT), alpha-L-fucosidase (AFU), alpha fetoprotein (AFP), carcino-embryonic antigen (CEA), carbohydrate antigen 19-9 (CA19-9), viral load and platelet counts. Statistical analysis was performed using the SPSS software system (SPSS 10.0). Results: In real-time PCR, fluorescence was detectable in all blood specimens from patients with HCC and healthy volunteers. The levels of TGF-β1 mRNA expression in patients with HCC were significantly higher compared to that in healthy volunteers (P<0.000 1), suggesting an association of the activated TGF-β1 gene transcription with hepato- carcinogenesis. Patients with HCC were divided into 2 groups according to their TGF-β1 mRNA above (group A, n=28)or below (group B, n=25) the mean level. Statistical results demonstrated that TGF-β1 mRNA expression level was correlated with patients age, serum levels of CEA, CA19-9 and viral copy number (P<0.05). Conclusion: Although this is a small sample size pilot study these findings imply that quantitative measurement of TGF-β1 mRNA level in peripheral blood may be a complementary serologic marker of HCC.展开更多
The aim of the paper was to study the metabolite profile and morphological characteristics of sugar beet regenerants exposed to aluminium ions (Al^3+). The regenerants were selected basing on selective media with s...The aim of the paper was to study the metabolite profile and morphological characteristics of sugar beet regenerants exposed to aluminium ions (Al^3+). The regenerants were selected basing on selective media with sublethal acidity (pH 3.5). The thrice-repeated passaging of sugar beet microclones of two genotypes in low pH medium causes certain alterations in the cellular metabolism. The paper demonstrated that peroxidase (POD) and isocitrate lyase (ICL) activity increased in both varieties. At the same time, NADH-dehydrogenase (NADH-DH) activity decreased in hybrid plants. Glucose-6-phosphate-dehydrogenase (gl-6-ph-dh) activity increased in mail sterile (MS) hybrid plants, but reduced in Ramonskaya fertile (RF) hybrid plants. Adaptation to reduced pH was accompanied by alterations in the isozyme spectra of POD, 1- and 2-esterase, cytochrome c oxidase and malic enzyme (ME). The adaptation process of sugar beet regenerants was also accompanied by an increase in protein synthesis. The level of metabolic response to stress very much depended on the initial genotype of the hybrid. In this experiment, aluminium resistant plants were growing rapidly in selective media. They developed leaves with healthy petioles and blades and had strong root systems.展开更多
Recent studies have demonstrated that Notch-1 expression is increased in the hippocampus of Alzheimer's disease patients. We speculate that Notch-1 signaling may be involved in PC12 cell apoptosis induced by amyloid ...Recent studies have demonstrated that Notch-1 expression is increased in the hippocampus of Alzheimer's disease patients. We speculate that Notch-1 signaling may be involved in PC12 cell apoptosis induced by amyloid beta-peptide (25-35) (Aβ25-35). In the present study, PC12 cells were cultured with different doses (0, 0.1, 1.0, 10 and 100 nmol/L) of N-[N-(3,5-Difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester, a Notch-1 signaling pathway inhibitor, for 30 minutes. Then cultured cells were induced with Aβ25-3s for 48 hours. Pretreatment of PC12 cells with high doses of N-[N-(3,5-Difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester (〉 10 nmol/L) prolonged the survival of PC12 cells after Aβ25-35 induction, decreased the expression of apoptosis-related proteins caspase-3, -8, -9, increased the activity of oxidative stress-related superoxide dismutase and catalase, inhibited the production of active oxygen, and reduced nuclear factor kappa B expression. This study indicates that the Notch-1 signaling pathway plays a pivotal role in Aβ25-35-induced PC12 apoptosis.展开更多
目的:研究幽门螺杆菌(Helicobacter pylori,H.pylori)对人肝正常细胞L-02(human normal liver cell line-02)的作用及转化生长因子β受体Ⅰ(transforming growth factor-beta typeⅠ,TβRⅠ)基因表达的影响.方法:体外培养L-02细胞、H.py...目的:研究幽门螺杆菌(Helicobacter pylori,H.pylori)对人肝正常细胞L-02(human normal liver cell line-02)的作用及转化生长因子β受体Ⅰ(transforming growth factor-beta typeⅠ,TβRⅠ)基因表达的影响.方法:体外培养L-02细胞、H.pylori;PCR法鉴定CagA+H.pylori和CagA-H.pylori,采用不同浓度的CagA+H.pylori和CagA-H.pylori作用于L-02细胞24 h,并设不加H.pylori的L-02细胞为阴性对照组;MTT法检测L-02细胞抑制率,通过Real-time PCR法检测各组TβRⅠ基因的表达.结果:MTT法结果显示,随着CagA+H.pylori、CagA-H.pylori浓度升高,对L-02细胞的抑制作用增强,与阴性对照组相比,差别均有统计学意义(P<0.05);在同一浓度,CagA+H.pylori对细胞的抑制作用较CagA-H.pylori明显,两组抑制率(%)分别为101 CFU/mL组10.960±0.231 vs 4.470±0.289;102 CFU/mL组25.310±0.398 vs 5.510±0.168;103 CFU/mL组33.130±0.312 vs 10.330±0.213;104 CFU/mL组54.570±0.245 vs 17.120±0.309;105 CFU/mL组79.450±0.402 vs 25.830±0.337;106 CFU/mL组90.210±0.271 vs 32.350±0.178,各组比较均t<0.05;Real-time PCR法检测中,随着CagA+H.pylori、CagA-H.pylori浓度升高,TβRⅠ表达逐渐升高,在同一浓度,CagA+H.pylori促TβRⅠ表达作用较CagA-H.pylori明显,两组TβRⅠ相对表达量分别为101 CFU/mL组1.65±0.101 vs 1.110±0.110;102 CFU/mL组2.770±0.198 vs 1.200±0.203;103 CFU/mL组4.590±0.112 vs 1.590±0.134;104 CFU/mL组5.470±0.145 vs 1.990±0.331;105 CFU/mL组7.450±0.102 vs 2.650±0.268;106 CFU/mL组8.570±0.221 vs 4.570±0.161,各组比较均t<0.05.结论:H.pylori对人肝正常细胞L-02细胞具有抑制作用并与浓度相关,菌液浓度越高,抑制细胞增殖作用明显,CagA+H.pylori的抑制作用比CagA-H.pylori强.H.pylori对L-02细胞作用后TβRⅠ基因表达增高并与浓度相关,CagA+H.pylori的影响比CagA-H.pylori大.H.pylori对L-02细胞抑制作用的可能机制是通过TβRⅠ基因表达改变干扰转化生长因子-β1/Smads信号通路传导而影响其生长.展开更多
基金Supported by:the Medicine and Health Scientific Research Projects of Shandong Province,No. 2007HZ065
文摘BACKGROUND: L-3-n-butylphthalide (L-NBP) can inhibit phosphorylation of tau protein and reduce the neurotoxicity of beta-amyloid peptide 1-42 (Aβ1-42). OBJECTIVE: To observe the neuroprotective effects of L-NBP on caspase-3 and nuclear factor kappa-B (NF- K B) expression in a rat model of Alzheimer's disease. DESIGN, TIME AND SETTING: A cell experiment was performed at the Central Laboratory of Provincial Hospital affiliated to Shandong University between January 2008 and August 2008. MATERIALS: L-NBP (purity 〉 98%) was provided by Shijiazhuang Pharma Group NBP Pharmaceutical Company Limited. Aβ1-42, 3-[4,5-dimethylthiazolo-2]-2,5 iphenyltetrazolium bromide (MTT), and rabbit anti-Caspase-3 polyclonal antibody were provided by Cell Signaling, USA; goat anti-choactase and rabbit anti-NF- kB antibodies were provided by Santa Cruz, USA. METHODS: Primary cultures were generated from rat basal forebrain and hippocampal neurons at 17 or 19 days of gestation. The cells were assigned into five groups: the control group, the Aβ1-42 group (2 μmol/L), the Aβ1-42 + 0.1 μmol/L L-NBP group, the Aβ1-42 + 1 μ mol/L L-NBP group, and the Aβ1-42 + 10μmol/L L-NBP group. The neurons were treated with Aβ1-42 (2 μmol/L) alone or in combination with L-NBP (0.1, 1, 10 μmol/L) for 48 hours. Cells in the control group were incubated in PBS. MAIN OUTCOME MEASURES: Morphologic changes were evaluated using inverted microscopy, viability using the M-I-I- method, and the changes in caspase-3 and NF- k B expression using Western blot. RESULTS: Induction with Aβ1-42 for 48 hours caused cell death and soma atrophy, and increased caspase-3 and NF- K B expression (P 〈 0.05). L-NBP blocked these changes in cell morphology, decreased caspase-3 and NF- k B expression (P 〈 0.05), and improved cell viability, especially at the high dose (P 〈 0.05). CONCLUSION: AI3^-42 is toxic to basal forebrain and hippocampal primary neurons; L-NBP protects against this toxicity and inhibits the induction of caspase-3 and NF- K B expression.
文摘A pot experiment was conducted during winter growing season of 2014 at Homs Agriculture Research Center, General Commission for Scientific Researches (GCSAR), Syria. A factorial experiment arranged according to complete randomized block design with six replications was used. A combination of four levels of saline irrigation water (tap water, 2,000, 4,000 and 6,000 ppm), with three K levels (180, 360 and 540 ppm), was used to evaluate the effects of saline irrigation water and K enrichment on some growth attributes of two sugar beet varieties (Semper and Alligator). Results showed that all studied growth attributes, i.e., leaf area (LA), leaf number (LN), total dry matter (TDM) and net assimilation rate (NAR) were decreased under salinity stress conditions compared to the control, while K enrichment significantly increased some of the studied characters such as LA, TDM and NAR, but the differences in LN were apparent according to increase in K levels. The variety Semper surpassed significantly the variety Alligator in LA, TDM and NAR. Results also indicated a significant interaction between salinity×potassium enrichment, varieties×potassium enrichment and salinity ? varieties.
基金the National Natural Science Foundation of China (30770994)
文摘Background: The carcinogenesis of hepatocellular carcinoma (HCC) is a multi-factorial, multistep and complex process. Its prognosis is poor and early detection is of the utmost importance. Transforming growth factor β1 (TGF-β1) message RNA (mRNA) has been reported to be elevated in HCC patients using Northern blotting. However, little work has been done about the detection of TGF-β1 mRNA levels in peripheral blood of patients with HCC using the real-time polymerase chain reactions (PCR) method. Objective: To assess the prognostic value of quantitative levels of TGF-β1 mRNA in peripheral blood of patients with HCC, and to investigate the relationship between the expression of TGF-β1 mRNA in peripheral blood and many diagnostic and pathological factors. Methods: We developed an optimized Taqman real-time PCR to quantify TGF-β1 mRNA in peripheral blood of 53 patients with HCC and 44 healthy volunteers. In addition, blood was collected from patients with HCC for measuring levels of total bilirubin (TBil), prealbumin, albumin, alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma glutamyltranspeptidase (GGT), alpha-L-fucosidase (AFU), alpha fetoprotein (AFP), carcino-embryonic antigen (CEA), carbohydrate antigen 19-9 (CA19-9), viral load and platelet counts. Statistical analysis was performed using the SPSS software system (SPSS 10.0). Results: In real-time PCR, fluorescence was detectable in all blood specimens from patients with HCC and healthy volunteers. The levels of TGF-β1 mRNA expression in patients with HCC were significantly higher compared to that in healthy volunteers (P<0.000 1), suggesting an association of the activated TGF-β1 gene transcription with hepato- carcinogenesis. Patients with HCC were divided into 2 groups according to their TGF-β1 mRNA above (group A, n=28)or below (group B, n=25) the mean level. Statistical results demonstrated that TGF-β1 mRNA expression level was correlated with patients age, serum levels of CEA, CA19-9 and viral copy number (P<0.05). Conclusion: Although this is a small sample size pilot study these findings imply that quantitative measurement of TGF-β1 mRNA level in peripheral blood may be a complementary serologic marker of HCC.
文摘The aim of the paper was to study the metabolite profile and morphological characteristics of sugar beet regenerants exposed to aluminium ions (Al^3+). The regenerants were selected basing on selective media with sublethal acidity (pH 3.5). The thrice-repeated passaging of sugar beet microclones of two genotypes in low pH medium causes certain alterations in the cellular metabolism. The paper demonstrated that peroxidase (POD) and isocitrate lyase (ICL) activity increased in both varieties. At the same time, NADH-dehydrogenase (NADH-DH) activity decreased in hybrid plants. Glucose-6-phosphate-dehydrogenase (gl-6-ph-dh) activity increased in mail sterile (MS) hybrid plants, but reduced in Ramonskaya fertile (RF) hybrid plants. Adaptation to reduced pH was accompanied by alterations in the isozyme spectra of POD, 1- and 2-esterase, cytochrome c oxidase and malic enzyme (ME). The adaptation process of sugar beet regenerants was also accompanied by an increase in protein synthesis. The level of metabolic response to stress very much depended on the initial genotype of the hybrid. In this experiment, aluminium resistant plants were growing rapidly in selective media. They developed leaves with healthy petioles and blades and had strong root systems.
文摘Recent studies have demonstrated that Notch-1 expression is increased in the hippocampus of Alzheimer's disease patients. We speculate that Notch-1 signaling may be involved in PC12 cell apoptosis induced by amyloid beta-peptide (25-35) (Aβ25-35). In the present study, PC12 cells were cultured with different doses (0, 0.1, 1.0, 10 and 100 nmol/L) of N-[N-(3,5-Difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester, a Notch-1 signaling pathway inhibitor, for 30 minutes. Then cultured cells were induced with Aβ25-3s for 48 hours. Pretreatment of PC12 cells with high doses of N-[N-(3,5-Difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester (〉 10 nmol/L) prolonged the survival of PC12 cells after Aβ25-35 induction, decreased the expression of apoptosis-related proteins caspase-3, -8, -9, increased the activity of oxidative stress-related superoxide dismutase and catalase, inhibited the production of active oxygen, and reduced nuclear factor kappa B expression. This study indicates that the Notch-1 signaling pathway plays a pivotal role in Aβ25-35-induced PC12 apoptosis.
文摘目的:研究幽门螺杆菌(Helicobacter pylori,H.pylori)对人肝正常细胞L-02(human normal liver cell line-02)的作用及转化生长因子β受体Ⅰ(transforming growth factor-beta typeⅠ,TβRⅠ)基因表达的影响.方法:体外培养L-02细胞、H.pylori;PCR法鉴定CagA+H.pylori和CagA-H.pylori,采用不同浓度的CagA+H.pylori和CagA-H.pylori作用于L-02细胞24 h,并设不加H.pylori的L-02细胞为阴性对照组;MTT法检测L-02细胞抑制率,通过Real-time PCR法检测各组TβRⅠ基因的表达.结果:MTT法结果显示,随着CagA+H.pylori、CagA-H.pylori浓度升高,对L-02细胞的抑制作用增强,与阴性对照组相比,差别均有统计学意义(P<0.05);在同一浓度,CagA+H.pylori对细胞的抑制作用较CagA-H.pylori明显,两组抑制率(%)分别为101 CFU/mL组10.960±0.231 vs 4.470±0.289;102 CFU/mL组25.310±0.398 vs 5.510±0.168;103 CFU/mL组33.130±0.312 vs 10.330±0.213;104 CFU/mL组54.570±0.245 vs 17.120±0.309;105 CFU/mL组79.450±0.402 vs 25.830±0.337;106 CFU/mL组90.210±0.271 vs 32.350±0.178,各组比较均t<0.05;Real-time PCR法检测中,随着CagA+H.pylori、CagA-H.pylori浓度升高,TβRⅠ表达逐渐升高,在同一浓度,CagA+H.pylori促TβRⅠ表达作用较CagA-H.pylori明显,两组TβRⅠ相对表达量分别为101 CFU/mL组1.65±0.101 vs 1.110±0.110;102 CFU/mL组2.770±0.198 vs 1.200±0.203;103 CFU/mL组4.590±0.112 vs 1.590±0.134;104 CFU/mL组5.470±0.145 vs 1.990±0.331;105 CFU/mL组7.450±0.102 vs 2.650±0.268;106 CFU/mL组8.570±0.221 vs 4.570±0.161,各组比较均t<0.05.结论:H.pylori对人肝正常细胞L-02细胞具有抑制作用并与浓度相关,菌液浓度越高,抑制细胞增殖作用明显,CagA+H.pylori的抑制作用比CagA-H.pylori强.H.pylori对L-02细胞作用后TβRⅠ基因表达增高并与浓度相关,CagA+H.pylori的影响比CagA-H.pylori大.H.pylori对L-02细胞抑制作用的可能机制是通过TβRⅠ基因表达改变干扰转化生长因子-β1/Smads信号通路传导而影响其生长.