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实时荧光定量PCR检测巨细胞病毒两种白细胞提取方法对比分析 被引量:3
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作者 刘广印 周光庭 沈佐君 《淮海医药》 CAS 2007年第2期95-96,共2页
目的比较实时荧光定量PCR技术检测巨细胞病毒两种白细胞提取处理方法。方法对51例新生儿黄疸患者外周血分别进行淋巴细胞分离液法提取白细胞和0.8%氯化胺裂解红细胞法提取白细胞,然后同时进行实时荧光定量PCR检测。结果51例样本中两种... 目的比较实时荧光定量PCR技术检测巨细胞病毒两种白细胞提取处理方法。方法对51例新生儿黄疸患者外周血分别进行淋巴细胞分离液法提取白细胞和0.8%氯化胺裂解红细胞法提取白细胞,然后同时进行实时荧光定量PCR检测。结果51例样本中两种方法均阳性5例,白细胞抽提液法阳性、氯化胺裂解红细胞法阴性2例;白细胞抽提液法阴性、氯化胺裂解红细胞法阳性4例;两者均阴性40例;氯化胺裂解红细胞阳性率法17.64%,淋巴细胞分离液法阳性率13.72%。经统计学分析,两方法阳性率差异无显著性(χ2=0.5,P>0.05)。结论氯化胺裂解红细胞法提取白细胞方法简便、可靠,可用于荧光定量PCR检测巨细胞病毒。 展开更多
关键词 实时荧光定量PCR 细胞病毒 淋巴细胞分离液法 氯化胺裂解红细胞
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黄芪甲苷通过抑制氯化物细胞内通道蛋白4/ADP核糖基化因子6信号通路改善血管紧张素Ⅱ诱导的高血压大鼠模型的心肌损伤 被引量:3
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作者 巩莎 可海霞 《中华高血压杂志》 CAS CSCD 北大核心 2024年第1期43-50,共8页
目的探究黄芪甲苷在血管紧张素Ⅱ(AngⅡ)诱导的高血压大鼠模型中的心肌保护作用及其机制。方法将60只SD大鼠分随机分为6组,每组10只:对照组,模型组,黄芪甲苷低(10 mg/kg)、中(20 mg/kg)、高剂量(50 mg/kg)组和ADP核糖基化因子6(Arf6)特... 目的探究黄芪甲苷在血管紧张素Ⅱ(AngⅡ)诱导的高血压大鼠模型中的心肌保护作用及其机制。方法将60只SD大鼠分随机分为6组,每组10只:对照组,模型组,黄芪甲苷低(10 mg/kg)、中(20 mg/kg)、高剂量(50 mg/kg)组和ADP核糖基化因子6(Arf6)特异性抑制剂(NAV-2729)组。建立AngⅡ诱导高血压心肌损伤大鼠模型后给予不同药物或剂量处理2周后,观察各组大鼠左心室舒张末期内径、心脏射血分数及短轴缩短率、心肌病理损伤、心肌细胞凋亡以及相关凋亡蛋白B细胞淋巴瘤因子-2(Bcl-2)及Bcl-2关联X蛋白(Bax)表达情况。另选30只SD大鼠随机分为3组,每组10只:氯化物细胞内通道蛋白4(CLIC4)过表达联合黄芪甲苷组、腺病毒阴性对照联合黄芪甲苷组及腺病毒阴性对照组。先通过尾静脉注射相应腺病毒,再用AngⅡ诱导高血压模型。2周后观察大鼠左心室舒张末期内径、心脏射血分数及短轴缩短率、心肌病理损伤、心肌细胞凋亡情况。结果与模型组大鼠比较,中剂量和高剂量的黄芪甲苷能够改善AngⅡ诱导的高血压心肌损伤模型大鼠的心脏功能及心肌结构,促进抗凋亡蛋白Bcl-2表达(0.36±0.01比0.22±0.02,0.78±0.01比0.22±0.02;均P<0.05),抑制Bax表达(1.51±0.02比2.13±0.03,1.22±0.01比2.13±0.03;均P<0.05)。黄芪甲苷通过抑制CLIC4/Arf6蛋白表达发挥心肌保护作用,过表达CLIC4则减弱这种保护作用。结论黄芪甲苷能够改善AngⅡ诱导的高血压大鼠模型中的心肌损伤,可能通过抑制CLIC4/Arf6信号通路实现。 展开更多
关键词 黄芪甲苷 氯化细胞内通道蛋白4/ADP核糖基化因子6 血管紧张素Ⅱ 高血压 心肌损伤
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Involvement of regulatory volume decrease in the migration of nasopharyn-geal carcinoma cells 被引量:9
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作者 JianWenMAO LiWeiWANG +7 位作者 TimJACOB XueRongSUN HuiLI LinYanZHU PanLI PingZHONG SiHuaiNIE LiXinCHEN 《Cell Research》 SCIE CAS CSCD 2005年第5期371-378,共8页
The transwell chamber migration assay and CCD digital camera imaging techniques were used to investigate the relationship between regulatory volume decrease (RVD) and cell migration in nasopharyngeal carcinoma cells (... The transwell chamber migration assay and CCD digital camera imaging techniques were used to investigate the relationship between regulatory volume decrease (RVD) and cell migration in nasopharyngeal carcinoma cells (CNE-2Z cells). Both migrated and non-migrated CNE-2Z cells, when swollen by 47% hypotonic solution, exhibited RVD which was inhibited by extracellular application of chloride channel blockers adenosine 5’-triphosphate (ATP), 5-nitro-2-(3- phenylpropylamino) benzoic acid (NPPB) and tamoxifen. However, RVD rate in migrated CNE-2Z cells was bigger than that of non-migrated cells and the sensitivity of migrated cells to NPPB and tamoxifen was higher than that of non- migrated cells. ATP, NPPB and tamoxifen also inhibited migration of CNE-2Z cells. The inhibition of migration was positively correlated to the blockage of RVD, with a correlation coefficient (r) = 0.99, suggesting a functional relation- ship between RVD and cell migration. We conclude that RVD is involved in cell migration and RVD may play an important role in migratory process in CNE-2Z cells. 展开更多
关键词 tumor cells chloride channel blockers cell volume cell migration transwell chamber.
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Differential regulation of survivin by p53 contributes to cell cycle dependent apoptosis 被引量:21
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作者 YanJIN YongWEI LeiXIONG YingYANG JiaRuiWU 《Cell Research》 SCIE CAS CSCD 2005年第5期361-370,共10页
Recent studies indicate that cell-cycle checkpoints are tightly correlated with the regulation of apoptosis, in which p53 plays an important role. Our present works show that the expression of E6/E7 oncogenes of human... Recent studies indicate that cell-cycle checkpoints are tightly correlated with the regulation of apoptosis, in which p53 plays an important role. Our present works show that the expression of E6/E7 oncogenes of human papillomavirus in HeLa cells is inhibited in the presence of anti-tumor reagent tripchlorolide (TC), which results in the up-regulation of p53 in HeLa cells. Interestingly, under the same TC-treatment, the cells at the early S-phase are more susceptible to apoptosis than those at the middle S-phase although p53 protein is stabilized to the same level in both situations. Significant difference is exhibited between the two specified expression profiles. Further analysis demonstrates that anti-apoptotic gene survivin is up-regulated by p53 in the TC-treated middle-S cells, whereas it is down-regulated by p53 in the TC-treated early-S cells. Taken together, the present study indicates that the differential p53-regulated expres- sion of survivin at different stages of the cell cycle results in different cellular outputs under the same apoptosis-inducer. 展开更多
关键词 APOPTOSIS cell cycle P53 SURVIVIN HeLa cell.
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Negundoside,an irridiod glycoside from leaves of Vitex negundo,protects human liver cells against calcium-mediated toxicity induced by carbon tetrachloride 被引量:2
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作者 Sheikh A Tasduq Peerzada J Kaiser +2 位作者 Bishan D Gupta Vijay K Gupta Rakesh K Johri 《World Journal of Gastroenterology》 SCIE CAS CSCD 2008年第23期3693-3709,共17页
AIM: To evaluate the protective effect of 2′-p-hydroxy benzoylmussaenosidic acid [negundoside (NG), against carbon tetrachloride (CCl4)-induced toxicity in HUH-7 cel Is.METHODS: CCI4 is a well characterized hep... AIM: To evaluate the protective effect of 2′-p-hydroxy benzoylmussaenosidic acid [negundoside (NG), against carbon tetrachloride (CCl4)-induced toxicity in HUH-7 cel Is.METHODS: CCI4 is a well characterized hepatotoxin, and inducer of cytochrome P450 2E1 (CYP2E1)-mediated oxidative stress. In addition, lipid peroxidation and accumulation of intracellular calcium are important steps in the pathway involved in CCl4 toxicity. Liver cells (HUH-7) were treated with CCI4, and the mechanism of the cytoprotective effect of NG was assessed. Silymarin, a known hepatoprotective drug, was used as control. RESULTS: NG protected HUH-7 cells against CCl4 toxicity and loss of viability without modulating CYP2E1 activity. Prevention of CCl4 toxicity was associated with a reduction in oxidative damage as reflected by decreased generation of reactive oxygen species (ROS), a decrease in lipid peroxidation and accumulation of intracellular Ca^2+ levels and maintenance of intracellular glutathione homeostasis. Decreased mitochondrial membrane potential (MMP), induction of caspases mediated DNA fragmentation and cell cycle arrest, as a result of CCl4 treatment, were also blocked by NG. The protection afforded by NG seemed to be mediated by activation of cyclic adenosine monophosphate (cAMP) synthesis and inhibition of phospholipases (cPLA2). CONCLUSION: NG exerts a protective effect on CYP2E1-dependent CCl4 toxicity via inhibition of lipid peroxidation, followed by an improved intracellular calcium homeostasis and inhibition of Ca^2+-dependent proteases. 展开更多
关键词 Negundoside SILYMARIN HUH-7 Carbon tetrachloride CYP 2E1 Oxidative stress Calcium TOXICITY
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Photoprotective Effects of Hydroxychloroqine and TCMs on Human Keratinocytes Damaged from Ultraviolet Irradiation 被引量:10
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作者 LUODan MINWei LINXiang-fei WUDi MIAOXu 《Journal of Nanjing Medical University》 2004年第4期172-177,共6页
Objective: To investigate damage effects of ultraviolet irradiation on eternal keratinocyte-HaCaT cells and to evaluate photo-protective efficiency of hydroxychloroqine and Traditional Chinese Medicines(epigallocatech... Objective: To investigate damage effects of ultraviolet irradiation on eternal keratinocyte-HaCaT cells and to evaluate photo-protective efficiency of hydroxychloroqine and Traditional Chinese Medicines(epigallocatechingallate[EGCG], baikal skullcap root and szechwan lovge rhizome) on HaCaT cells damaged by middle wave ultraviolet(UVB) irradiation. Methods: Subconfluent HaCaT cells were sham or UVB irradiated and treated with above TCM agents. The damage degree of HaCaT cells was observed by a light microscop. Cell growth was recorded by cell count and cellular activity was detected by MTT method. The secretion amount of IL-6 and TNF-α was measured by ELISA. Results: The irradiation damage of HaCaT cells was depended on the irradiated dosages and cellular activity was reduced by 36%-80%, with a maximum decrease over 90% after 72 h. The intervention of the above drugs may increase the cellular activity by 10%-72%. The photo-protective efficiency was more apparent in EGCG (from 1.19±0.07 to 1.28±0.06, P<0.01) than that in hydroxychloroqine (from 0.43±0.04 to 0.96±0.04, P<0.05). The other two tested drugs also showed photo-protective effect(from 0.44±0.07 to 1.21±0.02, P<0.05). As to cytokine secretion, EGCG could decline the secretion amount of IL-6 and TNF-α apparently. Hydroxychloroqine and baikal skullcap root baikal skullcap root could only reduce the secretion of IL-6. The secretion of IL-6 and TNF-α could not be inhibited by szechwan lovge rhizome. Conclusion: The injury effect of UVB irradiation on cultured keratinocytes is dose-dependent and the tested drugs have photo-protective potency. Inhibition of cytokine secretion may be one of the mechanisms related to reducing the damage effect of UVB irradiation. 展开更多
关键词 ultraviolet irradiation photodamage and photoprotection KERATINOCYTES hydroxychloroqine Traditional Chinese Medicine baikal skullcap root szechwan lovge rhizome epigallocatechingallate
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Therapeutic effect of interleukin-10 on CCI_4-induced hepatic fibrosis in rats 被引量:27
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作者 Yue-Hong Huang Hei-Na Shi Wei-Da Zheng Li-Juan Zhang Zhi-Xin Chen Xiao-Zhong Wang 《World Journal of Gastroenterology》 SCIE CAS CSCD 2006年第9期1386-1391,共6页
AIM: To study the therapeutic effect of exogenous interleuldn-10 on CCl4-induced hepatic fibrosis in rats and its passible mechanisms. METHODS: Fourty-seven SD rats were randomly divided into control group (group N... AIM: To study the therapeutic effect of exogenous interleuldn-10 on CCl4-induced hepatic fibrosis in rats and its passible mechanisms. METHODS: Fourty-seven SD rats were randomly divided into control group (group N) and CCl4-induced hepatic fibrosis model group (group C). After CCl4 was given for 9 wk, the model group was divided into three groups. Rats in group H were put to death immediately, rats in group T were treated with IL-10 for another three wk and then put to death, rats in group R recovered after three weeks and were then killed. The degree of hepatic fibrosis was measured by HE staining and histological activity index (HAI). Histological activity index (HAI), change of collagen types Ⅰ and Ⅲ were measured by Picrosirius staining. The expression of TNF-α, HHP-2 and TIMP-1 in liver tissue was measured by S-P immunohis tochemistry.RESULTS: CCl4- induced experimental rat hepatic fibrosis model was established successfully. The degree of hepatic fibrosis was markedly lower in group T than in groups H and R, and there was no difference between the two groups. The expression of collagen types I and III was significantly suppressed in group T and was slightly suppressed in groups H and R. The positive levels of TNF-α, HHP-2 and TIHP-1 in group H increased significantly compared to those in group N (P〈0.01). The positive signals decreased significantly in groups T and R (P〈0.01), but positive score was significantly lower in group T than in group R (P〈 0.01). CONCLUS10N: Exogenous IL-10 can reverse CCl4-induced hepatic fibrosis in rats. IL-10 may exert its reversible effects on hepatic fibrosis by blocking CCl4-induced inflammation, inhibiting expression of HHP-2 and TIMP-1 and promoting resolution of collagen types Ⅰ and Ⅲ. 展开更多
关键词 Animals Carbon Tetrachloride Collagen Type I Collagen Type III Immunohistochemistry INTERLEUKIN-10 Liver Liver Cirrhosis Male Matrix Metalloproteinase 2 RATS Rats Sprague-Dawley Tissue Inhibitor of Metalloproteinase-1 Tumor Necrosis Factor-alpha
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Repeated-batch Cultivation of Encapsulated Monascus purpureus by Polyelectrolyte Complex for Natural Pigment Production 被引量:4
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作者 LIU Jiefeng REN Yiran YAO Shanjing 《Chinese Journal of Chemical Engineering》 SCIE EI CAS CSCD 2010年第6期1013-1017,共5页
In general,productions of natural pigment in submerged microorganism culture were much less than that in solid-state fermentation,because the solid-state culture can provide a support carrier for the mycelium. To impr... In general,productions of natural pigment in submerged microorganism culture were much less than that in solid-state fermentation,because the solid-state culture can provide a support carrier for the mycelium. To improve natural pigment production,the cultivation of Monascus purpureus in submerged encapsulated cell was investigated. Monascus purpureus immobilized in polyelectrolyte complex(PEC) microcapsules,which were pre-pared by sodium cellulose sulphate(NaCS) and poly-dimethyl-diallyl-ammonium chloride(PDMDAAC),was a good substitute for submerged cell culture because it mimicked the solid-state environment. The repeated-batch process with encapsulated cells was studied in flasks and a bubble column. The results indicated that the bubble column was more suitable for the encapsulation culture than the shaking flasks because of its good mass transfer performance and minor shear stress on cells. Owing to the protection of the microcapsule's membrane,Monascus purpureus in microcapsules increased approximately three times over that in free cell culture with negligible cell leakage to the medium. The pigment production in the bubble column finally reached 3.82(OD500) ,which was two times higher than in free cell culture. In addition,the duration of each batch was shortened to 15% of that in free cell culture. 展开更多
关键词 MICROENCAPSULATION Monascus purpureus natural pigment polyelectrolyte complex NaCS-PDMDAAC capsule cell immobilization
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Histological Study on the Skin of Japanese Flounder Paralichthys olivaceus
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作者 ZHUJie ZHANGXiumei GAOTianxiang 《Journal of Ocean University of China》 SCIE CAS 2005年第2期145-151,共7页
Histological development of Japanese flounder Paralichthys olivaceus larval skin and ultrastructural difference of skin between reared normal and malpigmented Japanese flounder were studied with light microscopy (LM) ... Histological development of Japanese flounder Paralichthys olivaceus larval skin and ultrastructural difference of skin between reared normal and malpigmented Japanese flounder were studied with light microscopy (LM) and transmission electron microscopy (TEM). The results show that the skin develops slowly before the metamorphosis, while at the onset of metamorphosis, the skin develops quickly and becomes complete in structure till about 50 d after being hatched. Ultrastructural observation on the normal and malpigmented skins shows that the iridophore and melanophore are adjacent to each other. Profile and structure of the two kinds of pigment cells are more complete in the skin of normal ocular side than in the skin of pigmented blind side. The ultrastructure of typical chloride cell was observed in the skin of Japanese flounder larvae for the first time. 展开更多
关键词 HISTOLOGY SKIN Paralichthys olivaceus melanophore IRIDOPHORE chloridecell
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CYTOTOXICITY AND GENOTOXICITY OF POLYETHYLENIMINE AND NICKEL CHLORIDE IN RED SEA BREAM (Pagrosomus major) FIN CELL LINE RSBF 被引量:2
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作者 郭华荣 张士璀 《Chinese Journal of Oceanology and Limnology》 SCIE CAS CSCD 2002年第4期323-331,共9页
A continuous marine fish cell line RSBF (i.e. Red Sea Bream Fin) was utilized to screen the cytotoxicity and genotoxicity of polyethylenimine (PEI) and nickel chloride (NiCl 2) in this study on the deleterious effects... A continuous marine fish cell line RSBF (i.e. Red Sea Bream Fin) was utilized to screen the cytotoxicity and genotoxicity of polyethylenimine (PEI) and nickel chloride (NiCl 2) in this study on the deleterious effects of aquatic genotoxins on fish. At the 0.01 to 1 μg/ml concentration tested, PEI had acute toxicity to the treated RSBF cells (IC 50 =1.12, 0.92, 0.88 and 0.64 μg/ml PEI for time 0 h, 24 h, 48 h and 72 h after treatment, respectively) and markedly inhibited their proliferation in a dose dependent manner. At the 0.001 to 5 μmol/L concentration tested, NiCl 2 posed no acute toxicity but significantly stimulated their growth (107%-214% of control). Random amplified polymorphic DNA (RAPD) technique was used to detect the genotoxic effects of PEI and NiCl 2 by comparing the RAPD banding patterns of the control and treated cells. RAPD analysis indicated that at the concentrations tested, PEI was more genotoxic than NiCl 2 to RSBF cells; that there was a slight dose dependent response in the genotoxic effect of PEI but not NiCl 2; and that RAPD technique might provide a sensitive, non specific genotoxic endpoint. And the potent cytotoxicity and genotoxicity of PEI on fish cells showed that we should be cautious in utilizing it as gene vector in fish gene transfer and human gene therapy. 展开更多
关键词 fish cell line CYTOTOXICITY GENOTOXICITY polyethylenimine (PEI) nickel chloride (NiCl 2) random amplified polymorphic DNA (RAPD)
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Contribution of mononuclear bone marrow cells to carbon tetrachloride-induced liver fibrosis in rats 被引量:4
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作者 Bao-Qiang Cao Ji-Zong Lin +6 位作者 Yue-Si Zhong Shao-Bin Huang Nan Lin Zhao-Feng Tang Rui Chen Peng Xiang Rui-Yun Xu 《World Journal of Gastroenterology》 SCIE CAS CSCD 2007年第12期1851-1856,共6页
AIM: To study the inhibitory effect of mononuclear bone marrow cell (BNC) transplantation on carbon tetrachloride (CCl4) -induced liver fibrosis in rats. METHODS: Rat liver fibrosis models were induced by CCl4 a... AIM: To study the inhibitory effect of mononuclear bone marrow cell (BNC) transplantation on carbon tetrachloride (CCl4) -induced liver fibrosis in rats. METHODS: Rat liver fibrosis models were induced by CCl4 and alcohol administration. After 8 wk, twenty rats were randomly allocated into treatment group (n = 10) and control group (n = 10). BMC were infused into the rats in treatment group via the portal vein, while heparinized saline was infused in control group. CCl4 was hypodermically injected into the rats twice a week for 4 wk. At the end of wk 12, all rats were humanely sacrificed. Uver samples were taken and stained with HE or Masson trichrome. The general conditions, liver fibrosis (hydroxyproline and collagen fibre) and liver pathological grades in rats were evaluated. RESULTS: The general conditions of the rats in treatment group improved markedly, but not in control group. Hydroxyproline was 504.6± 128.8 μg/g in treatment group, and 596.0 ± 341.8 μg/g in control group. The percentage of collagen fibre was 3.75% ± 0.98% in treatment group and 5.02% ± 0.44% in control group. There was a significant difference between the two groups (P 〈 0.05). Liver pathological grade decreased from grade N to grade 11 partially in treatment group (P 〈 0.05) with no obvious improvement in control group (P 〉 0.05). There was a significant difference between treatment group and control group (P 〈 0.05).CONCLUSION: Transplantation of BMC can improve liver fibrosis due to chronic liver injury in rats. 展开更多
关键词 Bone marrow cell TRANSPLANTATION Liver cirrhosis Rat model
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维生素E抗VD_3所致心肌损伤的实验研究 被引量:1
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作者 郭文萃 曾宪惠 +3 位作者 迟月明 万汇娟 王丽宏 姚东升 《哈尔滨医科大学学报》 CAS 1995年第6期455-457,共3页
本文用大剂量VD_3复制心肌损伤模型,通过光镜、常规电镜、细胞化学及硝酸镧电子示踪进一步观察VD_3致心肌损伤的形态及VE对其保护作用。实验结果表明,VD_3能增加心肌细胞膜通透性,降低呼吸酶(细胞色素氧化酶和琥珀酸脱... 本文用大剂量VD_3复制心肌损伤模型,通过光镜、常规电镜、细胞化学及硝酸镧电子示踪进一步观察VD_3致心肌损伤的形态及VE对其保护作用。实验结果表明,VD_3能增加心肌细胞膜通透性,降低呼吸酶(细胞色素氧化酶和琥珀酸脱氢酶)活性。VE能改善膜的通透性,保护呼吸酶活性,明显减少心肌坏死面积,使心肌超微结构和功能的损伤有所改善。 展开更多
关键词 维生素D3 维生素E 细胞色素氯化 心肌损伤
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Ultrastructural Observation of the Skin Chloride Celis of Japanese Flounder Paralichthys olivaceus and Turbot Scophthamus maximus Larvae
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作者 朱杰 Zhang +2 位作者 Xiumei Gao Tianxiang 《High Technology Letters》 EI CAS 2003年第1期31-34,共4页
The ultrastructures of skin chloride celis in cultured Japanese flounder and turbot larvae in metamorphosis, which grow in the same feeding conditions, are examined with a transmission electron microscope. These devel... The ultrastructures of skin chloride celis in cultured Japanese flounder and turbot larvae in metamorphosis, which grow in the same feeding conditions, are examined with a transmission electron microscope. These developed skin chloride celis were shaped like flattened ellipsoids and similar in morphology and ultrastructure to typical chloride celis of euryhaline fish gill. They lo-cate in the epidermis and contract with the extra and interior environment through the apical pit and narrow channels. The cytoplasm of cell is full of numerous mitochondria and a ramifying net-work of tubules. The degeneration of skin chloride celis is observed with development of Japanese flounder larvae. Skin chloride celis of turbot are less developmental than those of Japanese flounder in the same developmental stage. 展开更多
关键词 ULTRASTRUCTURE skin chloride cell Paralichthys olivaceus Scophthamus maximus LARVAE
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Genotypic Assessment by RAPD Markers and Ultrastructural Characteristics of a NaCI-Tolerant Potato Cell Line
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作者 Filipa Queiros Jose M. Almeida +1 位作者 Domingos P.F. Almeida Femanda Fidalgo 《Journal of Life Sciences》 2012年第1期1-8,共8页
Salinity is a serious threat to agricultural production. Potato (Solanum tuberosum) is an important food crop characterised for having low to moderate salinity tolerance. Tissue cultures may be relevant to improve s... Salinity is a serious threat to agricultural production. Potato (Solanum tuberosum) is an important food crop characterised for having low to moderate salinity tolerance. Tissue cultures may be relevant to improve salt tolerance in potato through selection of salt-tolerant cell lines and subsequent regeneration of plants. In this work, the authors used the random amplified polymorphic DNA (RAPD) markers to investigate the occurrence of genetic polymorphism in a potato calli line tolerant to 150 mM NaCI. Out of 40 primers screened, eight generated polymorphic patterns that distinguished salt-tolerant line from the control. Although the macroscopic appearance was similar in both lines, ultrastructural study revealed alterations in salt-grown cells. These showed that plastids less differentiated with a lower number of grana had more and larger starch grains than control cells. In conclusion, RAPD analysis revealed that NaCl-adapted line is a somaclonal variant and the ultrastructural study showed changes essentially at the plastids. 展开更多
关键词 Salt tolerance POTATO callus tissue polymorphism molecular marker RAPD markers ultrastructure.
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ISOLATION OF HEPATIC OVAL CELLS FROM DIFFERENT MODEL RATS INCLUDING DIABETIC RATS 被引量:1
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作者 陆颖理 叶婷婷 +3 位作者 夏芳珍 王宁荐 杨华 陈奕 《Journal of Shanghai Second Medical University(Foreign Language Edition)》 2009年第1期7-11,共5页
Objective To acquire oval cells (progenitor stem cells ) from adult rat liver of different models including diabetic rats. Methods Thirty Sprague-Dawley ( SD ) rats were divided into 5 groups randomly: control, 2... Objective To acquire oval cells (progenitor stem cells ) from adult rat liver of different models including diabetic rats. Methods Thirty Sprague-Dawley ( SD ) rats were divided into 5 groups randomly: control, 2-acetylaminofluorene ( 2-AAF ), 2-AAF + partial hepatectomy ( PH ), 2-AAF + carbon tetrachloride ( CCl4 ), and diabetic groups. As two-step collagenase perfusion protocol of Seglen, oval cells were isolated by Percoll density gradient centrifugation. Thy1. 1 positive cells were sorted by flow cytometry, and then cultured in Dulbecco's minimum Eagle's medium (DMEM). Immunofluorescence staining was applied to labelling Thyl. 1. Results Different rates of Thy1.1 positive oval cells were found in different rat model groups : 0. 5 % in 2-AAF, 0. 3% in 2-AAF + PH, 0. 2% in 2-AAF + CCl4, 0. 1% in diabetic, and 0. 0% in control. Isolated cells adhered to plate with fusiform or polygon as epithelial cells. Conclusion Progenitor stem cells exist in injured liver tissue including those from diabetic rats. 展开更多
关键词 diabetes mellitus hepatic oval cells stem cells cell sorting
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Driving Forces of AIDS Pathogenesis:Massive CD4^+ T Lymphocyte Depletion and Abnormal Immune Activation
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作者 Chang LI Qin-xue HU 《Virologica Sinica》 SCIE CAS CSCD 2009年第6期501-508,共8页
The occurrence of massive CD4+ T cell depletion is one of the most prominent characteristics of human immunodeficiency virus type 1 (HIV-1) infection during acute phase, resulting in unrestorable destruction to the im... The occurrence of massive CD4+ T cell depletion is one of the most prominent characteristics of human immunodeficiency virus type 1 (HIV-1) infection during acute phase, resulting in unrestorable destruction to the immune system. The infected host undergoes an asymptomatic period lasting several years with low viral load and ostensibly healthy status, which is presumably due to virus-specific adaptive immune responses. In the absence of therapy, an overwhelming majority of cases develop to AIDS within 8-10 years of latent infection. In this review, we discuss the roles in AIDS pathogenesis played by massive CD4+ T lymphocytes depletion in gut-associated lymphoid tissue (GALT) during acute infection and abnormal immune activation emerging in the later part of chronic phase. 展开更多
关键词 HIV/AIDS CD4^+ T cell depletion Gut-associated lymphoid tissue Immune activation PATHOGENESIS
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Effect of Abscisic Acid on NaCI Stressed Callus Proliferation and Plant Regeneration in Rice
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作者 ikram-ul-Haq Ghulam Yasin +1 位作者 Mumtaz Hussain Ali Mohammad Dahri 《Journal of Life Sciences》 2012年第1期48-54,共7页
In rice, 2,4-Dichlorphenoxyacetic acid (MS2) has been considered best for callus induction and combination ofNAA, IAA, ABA (MS3) for somatic embryogenesis. Efficient plant regeneration was observed when MS basal s... In rice, 2,4-Dichlorphenoxyacetic acid (MS2) has been considered best for callus induction and combination ofNAA, IAA, ABA (MS3) for somatic embryogenesis. Efficient plant regeneration was observed when MS basal salts supplied with 3% sorbitol and 3% maltose as carbon sources without hormones (MSac). During this experiment, effect of sodium chloride (NaCl) and abscisic acid (ABA) was assessed on callus growth, plant regeneration and root induction efficiency of rice (Oryza sativa L.) varieties IR-6 and Basmati-370. Callus proliferation rate was highly decreased in both varieties on MS2b (100 mol.m3 NaCl + 0.5 mg'L1 ABA) than MS2a (100 mol'm3 NaCl) cultures significantly. Proline, glycine-betaine and reducing sugars were increased significantly in MS2a and MS2b callusing cultures. However, total proteins were decreased in MS2a, while slightly increased in MS2b. Maximum plant regeneration (9.42 ±0.54 and 10.67 ±0.50 plantlets.callus1) from somatic embryos was observed on MS4c in IR-6 and Basmati-370, while 1.56 ± 0.06 (IR-6) and 0.95 ±0.05 (Basmati-370) plantlets callus1 were observed on MSab (100 mol·m3 NaCl). No plant regeneration was observed on MS4b (100 mol·m3 NaCl + 0.5 mg·L-1 ABA) medium in both varieties. Inhibition of root induction efficiency was high in MSsb (100 mol·m3 NaCl + 0.5 mg·L-1 ABA) than MS5a (100 mol·m3 NaCl) in the stressed cultures (P 〈 0.05). In this experiment, it was concluded that ABA involved in somatic embryogenesis and elevation of NaCl stress, while it causes inhibition of cell's growth as well as its regeneration into plantlets from somatic embryos. 展开更多
关键词 Oryza sativa L. NACL ABA embryogenic callus somatic embryogenesis bio-contents.
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Protective effects of edaravone against cobalt chloride-induced apoptosis in PC12 cells 被引量:5
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作者 陈吉相 赵婷 黄丹霞 《Neuroscience Bulletin》 SCIE CAS CSCD 2009年第2期67-74,共8页
Objective To investigate the neuroprotective effects of edaravone (Eda) on cobalt chloride (CoCl2)-induced oxidative stress and apoptosis in cultured PC12 cells as well as the underlying mechanisms. Methods PC12 c... Objective To investigate the neuroprotective effects of edaravone (Eda) on cobalt chloride (CoCl2)-induced oxidative stress and apoptosis in cultured PC12 cells as well as the underlying mechanisms. Methods PC12 ceils impaired by COCl2 were used as the cell model of hypoxia. MTT (methyl thiazolyl tetrazolium) was used to assay the viability of the PC12 ceils exposed to Eda with gradient concentrations; Hochest 33258 stain assay was used to analyze the apoptosis ratio of the PC12 cells; Bcl-2 and Bax protein levels in PC12 cells were examined by western blotting. ROS level, the mitochondrial transmembrane potential and caspase-3 activity in each group were detected by spectrofluorometer. Results COCl2 treatment caused the loss of cell viability in PC12 cells, which was associated with the elevation of apoptotic rate, the formation of ROS and the disruption of mitochondrial transmembrane potential. COCl2 also significantly induced the upregulation of Bax/Bcl-2 ratio and the activation of caspase-3. In contrast, Eda significantly reversed these phenotypes, with its maximum protective effect at 0.1 μmol/L. Conclusion These results indicated that Eda could protect PC12 cells from CoCl2-induced cytotoxicity, and this protection might be ascribed to its anti-oxidative and anti-apoptotic activities. 展开更多
关键词 COCl2 PC12 cells EDARAVONE oxidative stress APOPTOSIS
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The involvement of signaling activation of protein kinase C in gadolinium chloride-induced cell survival and cell cycle progression in NIH3T3 cells 被引量:1
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作者 冯敏 李金霞 +4 位作者 马孝杰 范云周 吴竞轩 曾群 杨晓改 《Journal of Chinese Pharmaceutical Sciences》 CAS CSCD 2014年第11期772-777,共6页
In the present study, we investigated the activation of protein kinase C (PKC) family in mouse embryonic fibroblast NIH3T3 cells using gadolinium chloride as a representative lanthanide ion. With live cell imaging s... In the present study, we investigated the activation of protein kinase C (PKC) family in mouse embryonic fibroblast NIH3T3 cells using gadolinium chloride as a representative lanthanide ion. With live cell imaging system and confocal laser scanning microscopy, we found that the treatment of 50 μM GdCI3 promoted cell survival under the condition of serum-starvation. Moreover, better cell attachment and cytoskeleton reorganization were also observed. Additionally, GdC13 treatment resulted in the phosphorylation of PKC family at different time points. Furthermore, bisindolylmaleimide (a PKCpan inhibitor) could efficiently reduce the level of phosphorylated PKCpan (βIISer660), alleviating ERK activation induced by GdC13. This finding indicated that the PKC activation was involved in GdC13-induced MAPK/ERK signaling and thus might contribute to GdClβ-indueed cell cycle progression and cell survival. 展开更多
关键词 Gadolinium chloride Cell cycle Protein kinase C CYTOSKELETON ERK NIH3T3 cells
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Ability of multicellular salt glands in Tamarix species to secrete Na^+ and K^+ selectively 被引量:8
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作者 MA HaiYan TIAN ChangYan +1 位作者 FENG Gu YUAN JunFeng 《Science China(Life Sciences)》 SCIE CAS 2011年第3期282-289,共8页
The present study aimed to determine the mechanism of cation-selective secretion by multicellular salt glands.Using a hydroponic culture system,the secretion and accumulation of Na+ and K+ in Tamarix ramosissima and T... The present study aimed to determine the mechanism of cation-selective secretion by multicellular salt glands.Using a hydroponic culture system,the secretion and accumulation of Na+ and K+ in Tamarix ramosissima and T.laxa under different salt stresses (NaCl,KCl and NaCl+KCl) were studied.Additionally,the effects of salt gland inhibitors (orthovanadate,Ba2+,ouabain,tetraethylammonium (TEA) and verapamil) on Na+ and K+ secretion and accumulation were examined.Treatment with NaCl (at 0-200 mmol L 1 levels) significantly increased Na+ secretion,whereas KCl treatment (at 0-200 mmol L 1 levels) significantly increased K+ secretion.The ratio of secretion to accumulation of Na+ was higher than that of K+.The changes in Na+ and K+ secretion differed after adding different ions into the single-salt solutions.Addition of NaCl to the KCl solution (at 100 mmol L 1 level,respectively) led to a significant decrease in K+ secretion rate,whereas addition of KCl to the NaCl solution (at 100 mmol L 1 level,respectively) had little impact on the Na+ secretion rate.These results indicated that Na+ secretion in Tamarix was highly selective.In addition,Na+ secretion was significantly inhibited by orthovanadate,ouabain,TEA and verapamil,and K+ secretion was significantly inhibited by ouabain,TEA and verapamil.The different impacts of orthovanadate on Na+ and K+ secretion might be the primary cause for the different Na+ and K+ secretion abilities of multicellular salt glands in Tamarix. 展开更多
关键词 TAMARIX salt gland INHIBITOR ion secretion salt stress
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