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目的:表达与纯化果蝇中SR蛋白家族新成员Dxl6 N端变体.方法:用PCR方法扩增得到Dxl6 N端基因片段,经酶切亚克隆至pGEX-4T-1原核表达载体,在大肠杆菌BL21(DE3)中与GST融合表达,并用谷胱甘肽-sepharose4B亲和层析柱纯化融合蛋白.结果:表达产物以可溶形式经亲和层析柱纯化后获得相对分子质量约为37 000的融合蛋白.结论:成功克隆、表达并纯化了果蝇神经特异性拼接因子Dxl6N端变体与GST的融合蛋白. 相似文献
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目的:表达与纯化果蝇中SR蛋白家族新成员Dxl6N端变体。方法:用PCR方法扩增得到Dxl6N端基因片段,经酶切亚克隆至pGEX-4T-1原核表达栽体。在大肠杆菌BL21(DE3)中与GST融合表达。并用谷胱甘肽-sepharose4B亲和层析柱纯化融合蛋白。结果:表达产物以可溶形式经亲和层析柱纯化后获得相对分子质量约为37000的融合蛋白。结论:成功克隆、表达并纯化了果蝇神经特异性拼接因子Dxl6N端变体与GST的融合蛋白。 相似文献
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目的:探讨培哚普利对链脲佐菌素(STZ)诱导的大鼠糖尿病模型肾小管间质中清道夫受体A(SR‐A)表达的影响。方法将大鼠随机分为健康对照组、糖尿病组和培哚普利干预组。检测各组大鼠的血糖和尿清蛋白,行M asson染色观察肾脏形态学变化,应用逆转录‐聚合酶链反应(RT‐PCR)和免疫组织化学检测SR‐A mRNA及蛋白的表达。结果与健康对照组比较,糖尿病组大鼠血糖和24 h尿清蛋白均显著上升;肾组织SR‐A mRNA和蛋白表达均显著上调,培哚普利干预后,上述指标除血糖外均明显被抑制(P<0.05)。结论糖尿病大鼠肾小管间质SR‐A表达增加,培哚普利对糖尿病大鼠肾脏有保护作用,其机制可能部分与下调肾小管间质SR‐A表达有关。 相似文献
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一个新的人信号识别微粒受体基因APMCF1的克隆、表达和抗体制备 总被引:1,自引:1,他引:0
目的:克隆人类新基因APMCF1,原核表达并制备其特异性多克隆抗体.方法:运用5’末端快速扩增方法,结合Genbank数据库进行电子拼接,完善APMCF1的5’端序列;利用RT-PCR方法,扩增出APMCF1的蛋白编码cDNA序列,克隆入pGEX-KG中,构建APMCF1的原核表达载体,在大肠杆菌中诱导表达后获得了GST-APMCF1融合蛋白,以此融合蛋白为免疫原制备兔抗血清,用Western blot鉴定抗体特异性.结果:通过RACE及电子拼接,完善了APMCF1的5’端序列,其cDNA长度为1745bp,与鼠信号识别微粒受体β亚基序列同源性较高,染色体定位于3q22.2;克隆了APMC2F1 816bp的开放读框序列并表达了GST-APMCF1融合蛋白.制备了特异性兔抗GST-APMCF1多克隆抗体.结论:APMCF1可能是编码人类信号识别微粒受体β亚基的基因.该分子多克隆抗体的初步制备,为进一步研究APMCF1的功能提供了必要工具。 相似文献
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目的 观察大麻素1型受体拮抗剂SR141716A(SR1)对BV2小胶质细胞免疫调节功能的影响.方法 使用致炎因子干扰素-γ(IFN-γ)刺激激活BV2小胶质细胞,建立模拟EAE炎性环境的细胞模型,比较静息态BV2细胞组、激活态BV2细胞组和SR1干预组CB1R mRNA和蛋白的表达情况,用ELISA方法检测细胞因子和趋化因子的浓度,Griess试剂法检测一氧化氮(NO)浓度,MTT法检测细胞增殖率.结果 激活态的BV2小胶质细胞CB1R mRNA和蛋白的表达均高于静息态BV2细胞组(P<0.05);大麻素1型受体拮抗剂SR1可降低激活态的BV2细胞CB1R mRNA和蛋白的表达(P <0.05);SR1可显著上调IFN-γ、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)的水平,增加BV2小胶质细胞NO的释放(P<0.05),而显著下调IL-4、IL-17和MCP-1的浓度,对IL-10、IL-1β和CX3CL1无调节作用.SR1对IFN-γ活化的BV2小胶质细胞增殖无影响(P>0.05).结论 CB1R参与了小胶质细胞介导的炎症反应,CB1R在调节细胞因子网络平衡和NO分泌中发挥了一定的作用. 相似文献
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甲状旁腺激素相关蛋白对发育中的Meckel’s软骨作用的研究 总被引:1,自引:1,他引:0
目的观察SD大鼠下颌骨发育过程中甲状旁腺激素相关蛋白(PTHrP)在Meckel’s软骨上时空的变化,初步探讨PTHrP在Meckel’s软骨发育中的意义.方法用免疫组化染色法及原位杂交染色法检测E13-19dSD胎鼠的下颌骨发育过程中,PTHrP及PTHrPmRNA在Meckel’s软骨上的表达.结果 E13-15d在Meckel’s软骨前段PTHrP蛋白强阳性表达,在细胞核、胞浆均表达,在肥大软骨细胞核上表达更明显,而且PTHrPmRNA强阳性表达的位置与PTHrP蛋白一致.E17-19d Meckel’s软骨前段骨化,E17d在临近成骨处的Meckel’s软骨软骨细胞PTHrP蛋白弱阳性表达,且位于肥大软骨细胞核内,PTHrPmRNA阴性表达.E19dPTHrP临近成骨处蛋白染色阴性表达,PTHrPmRNA阴性表达;在前段远离骨化处PTHrPmRNA,PTHrP蛋白强阳性表达,细胞核上表达明显.结论 PTHrP在Meckel’s软骨前段软骨细胞中的表达存在时空特异变化,提示PTHrP蛋白可能由SD大鼠Meckel's软骨软骨细胞合成,并作为一种自分泌因子调节Meckel’s软骨细胞的增殖和分化. 相似文献
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目的 明确在慢性心房颤动(AF)中Junctophilin-2 (JPH2)表达变化及意义.方法 从心脏外科体外循环手术中获取21例窦性心律(SR)患者和30例慢性AF患者右心房肌组织,采用免疫组织化学方法测定心房肌组织JPH2蛋白的分布;实时荧光定量PCR法检测心房肌JPH2 mRNA水平;Western blot测定人心房肌JPH2蛋白水平.结果 免疫组织化学实验发现SR组与AF组心房肌组织上均存在JPH2蛋白表达,SR组着色更明显;AF组心房肌JPH2 mRNA水平和蛋白水平均明显低于SR组(P<0.01).结论 慢性AF患者心房肌JPH2的表达下调可能与心房肌细胞内钙紊乱密切相关. 相似文献
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Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P < 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (>0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P <0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P < 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD. 相似文献
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Jiageng Zhu Jun Li Ruipeng Ji Jianghao Su Mingshun Shen Zhigang Cao 《南京医科大学学报(自然科学版)》2007,21(2):125-128
Objective: To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods: SD rats served as both donors and recipients. 4℃ sodium lactate Ringer's was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava (IVC) inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results: Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion: This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment. 相似文献
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Tao Ye * Zhi-quan Liu Jian-jun Mu Xi-han Fu Jun Yang Bao-lin Gao and Xiao-hong Zhang Department of Cardiology First Hospital of Xi'an Jiaotong University Xi'an Hanzhong Cardiovascular Institute Hanzhong 《中国医学科学杂志(英文版)》2004,19(4):248-251
Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P< 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P< 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure. 相似文献
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目的:评价使用安心颗粒对急诊经皮冠状动脉介入术(PPCI)术后生活质量的影响.方法:将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组(术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次)和对照组(仅接受基础药物治疗).所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表(MIDAS)、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果:入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异(P>0.05);出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低(P<0.05);组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低(P<0.05).两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血(P>0.05).两组发生轻度血小板减少症的患者数比较无差异(P>0.05).结论:PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险. 相似文献
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Jingming Zhu Ningning Hui Xiuhong Jiang Huaxing Zhang Chengzhang Fu 《南京医科大学学报(自然科学版)》2007,21(6):386-389
Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P<0.01);the MAP decreased(P<0.01)and the HR increased drastically(P<0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients. 相似文献
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Objective:To investigate the gene expression of osteoprotegerin(OPG) and osteoclast differentiation factor(ODF) in the bone tissue of patients with hip fracture due to osteoporosis. Methods:OPGmRNA and ODFmRNA in the bone tissue in 50 cases of osteoporosis sufferers(over 50 years old) with hip fracture(Observer Group) and 30 cases of hip facture sufferers with no osteoporosis(Control group) were analyzed with the Semi-Quantitative RT-PCR method. Results:The mRNA expressed of ODF, OPG were both high in the patients with hip fracture. In the control group, the expression of OPG mRNA was observed, while the expression of ODF mRNA was very slight. Conclusion:Aged patients contained all signals including OPG, ODF that are essential for inducing osteoclastogenesis and promoting bone resorption. 相似文献
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Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P. 相似文献
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Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine. 相似文献
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Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center. 相似文献
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Scientific principles and rigorous processes should be followed in developing clinical guidelines for therapeutic interventions of integrative medicine 下载免费PDF全文
A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care(~'4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes. 相似文献