抗纤维化短肽对血清诱导的心脏成纤维细胞增殖和胶原合成与Ⅰ、Ⅲ型胶原表达抑制作用的实验研究

①目的 探讨N-乙酰基-丝氨酰-天门冬酰-赖氨酰-脯氨酸(AcSDKP)对不同浓度血清(FBS)诱导的大鼠心脏成纤维细胞增殖和胶原合成的调节作用.②方法 选用新生大鼠心脏成纤维细胞;采用3H-TdR掺入法检测心脏成纤维细胞的增殖;采用3H-脯氨酸掺入法检测心脏成纤维细胞胶原的合成,采用westernblot法检测心脏成纤维细胞Ⅰ型与Ⅲ型胶原的表达.③结果 在0.4%、2%和10%几种血清浓度的条件下,随着血清农度的增加,反映心脏成纤维细胞增殖3H-TdR掺入量和胶原合成的3H-脯氨酸掺入量增加.表明一定浓度的血清能诱导心脏成纤维细胞增殖和胶原合成.与0.4%血清的基础培养条件相比较,10%高浓度血清能刺激心脏成纤维细胞Ⅰ、Ⅲ型胶原蛋白的表达.在10-10～10-8mol/L浓度范围内.AcSDKP对10%诱导的心脏成纤维细胞增殖和胶原合成有抑制作用.并且在10-9mol/L时,AcSDKP对心脏成纤维细胞增殖、胶原合成抑制作用最强.同时10-9mol/L的AcSDKP对心脏成纤维细胞Ⅰ、Ⅲ型胶原蛋白的表达也有明显的抑制作用.④结论 AcSDKP对血清介导的心脏成纤维细胞增殖、胶原合成以及Ⅰ、Ⅲ型胶原蛋白的表达有明显抑制作用.     

干扰素α对肝纤维化鼠星状细胞Ⅰ和Ⅲ型前胶原mRNA表达及胶原沉积的影响

目的　观察干扰素α对大鼠肝纤维化时星状细胞增殖、Ⅰ、Ⅲ型前胶原mRNA表达和肝脏胶原沉积的影响。方法　以CCl4制造肝纤维化模型 ,培养肝星状细胞 ,抽提RNA ,用地高辛标记Ⅰ、Ⅲ型前胶原和胶原酶cDNA探针 ,Northern杂交分析Ⅰ、Ⅲ型前胶原和胶原酶mRNA表达 ,Dotblot测定大鼠肝Ⅰ、Ⅲ型胶原沉积。分别用3H TdR和3H 脯氨酸掺入观察干扰素α对星状细胞增殖和胶原合成的影响。结果　干扰素α对3H TdR和3H 脯氨酸掺入的星状细胞 ,在 4、2 4、4 8小时与空白对照比较 ,呈明显抑制作用 ,其抑制率分别为 16%、19%、2 7%。干扰素α浓度≥ 12 5U/ml时呈现明显抑制作用 ,且随浓度增加抑制作用加强。在鼠肝纤维化早、中、晚期 ,干扰素α组星状细胞Ⅰ、Ⅲ型前胶原mRNA表达与模型对照组比较均降低。星状细胞间质胶原酶mRNA表达 :在模型对照组早、中期均高于正常对照组 ,而晚期与正常对照组差异无显著意义 ;在干扰素α组早、中、晚各期 ,星状细胞间质胶原酶mRNA表达与模型对照组差异无显著意义。肝内Ⅰ、Ⅲ型胶原沉积 :干扰素α组早、中期均低于模型对照组 ,而晚期与模型对照组差异无显著意义。结论　干扰素α可抑制星状细胞的增殖和胶原合成 ,下调肝纤维时Ⅰ、Ⅲ型前胶原mRNA表达 ,减少肝脏Ⅰ、Ⅲ型胶原沉积。     

高糖状态下心肌成纤维细胞Ⅰ、Ⅲ型胶原表达变化的观察

目的探讨高糖对大鼠心肌成纤维细胞Ⅰ、Ⅲ型胶原表达的影响。方法采用胰酶消化法和差速贴壁分离法获取心肌成纤维细胞,采用正常糖(5.5mmol/L,对照组)和高糖(25mmol/L,高糖组)干预。用实时荧光定量PCR(Real-time PCR)方法检测Ⅰ、Ⅲ型前胶原mRNA表达,酶联免疫吸附测定法(ELISA)观察Ⅰ、Ⅲ型胶原蛋白水平的变化。结果与对照组比较,高糖刺激12h可使心肌成纤维细胞Ⅰ、Ⅲ型前胶原mRNA表达增加,Ⅰ、Ⅲ前胶原mRNA表达在12h升高,在24h达高峰,Ⅰ型前胶原mRNA升高幅度高于Ⅲ型前胶原mRNA;高糖刺激48h可使Ⅰ、Ⅲ型胶原蛋白表达增加,Ⅰ型胶原蛋白升高幅度高于Ⅲ型胶原。结论高糖可促进大鼠心肌成纤维细胞合成Ⅰ、Ⅲ型胶原。     

抗纤维化短肽对血清诱导的心脏成纤维细胞增殖和胶原合成与I、III型胶原表达抑制作用的实验研究

①目的探讨N-乙酰基-丝氨酰-天门冬酰-赖氨酰-脯氨酸(AcSDKP)对不同浓度血清(FBS)诱导的大鼠心脏成纤维细胞增殖和胶原合成的调节作用。②方法选用新生大鼠心脏成纤维细胞;采用3H-TdR掺入法检测心脏成纤维细胞的增殖;采用3H-脯氨酸掺入法检测心脏成纤维细胞胶原的合成,采用westernblot法检测心脏成纤维细胞I型与III型胶原的表达。③结果在0.4%、2%和10%几种血清浓度的条件下,随着血清浓度的增加,反映心脏成纤维细胞增殖3H-TdR掺入量和胶原合成的3H-脯氨酸掺入量增加。表明一定浓度的血清能诱导心脏成纤维细胞增殖和胶原合成。与0.4%血清的基础培养条件相比较,10%高浓度血清能刺激心脏成纤维细胞I、III型胶原蛋白的表达。在10-10~10-8mol/L浓度范围内,AcSDKP对10%诱导的心脏成纤维细胞增殖和胶原合成有抑制作用,并且在10-9mol/L时,AcSDKP对心脏成纤维细胞增殖、胶原合成抑制作用最强。同时10-9mol/L的AcSDKP对心脏成纤维细胞I、III型胶原蛋白的表达也有明显的抑制作用。④结论AcSDKP对血清介导的心脏成纤维细胞增殖、胶原合成以及I、III型胶原蛋...     

YC-1对低氧诱导的大鼠肺动脉外膜成纤维细胞增殖及胶原合成的影响

目的初步探讨低氧诱导因子抑制剂3-(5'-羟基甲基-2'-呋喃基)-1-苯基吲哚(YC-1)对低氧诱导的大鼠肺动脉外膜成纤维细胞增殖和胶原生成的影响,以及可能的分子机制。方法在低氧条件下培养的大鼠肺动脉成纤维细胞,将细胞随机分为常氧组、低氧组、低氧+YC-1(0.01、0.05和0.1 mmol/L)组。分别采用MTT法检测细胞增殖情况,3~H-脯氨酸掺入法测定胶原合成,蛋白免疫印迹法检测低氧诱导因子-1α(HIF-1α)蛋白的表达,逆转录-聚合酶链反应检测转化生长因子-β1(TGF-β1)mRNA表达情况。结果低氧组的成纤维细胞增殖及3~H-脯氨酸掺入量显著高于常氧组(均P0.01)。YC-1组的成纤维细胞增殖及3~H-脯氨酸掺入量显著低于低氧组,但仍高于常氧组;低氧组HIF-1α蛋白、TGF-β1 mRNA的表达明显高于常氧组(P0.01),YC-1呈剂量依赖性抑制缺氧刺激的HIF-1α蛋白、TGF-β1 mRNA表达,其中YC-1 0.1 mmol/L组中HIF-1α蛋白及TGF-β1 mRNA的表达分别抑制了65%和61%(均P0.01)。结论 YC-1能抑制低氧诱导的大鼠肺动脉外膜成纤维细胞增殖及胶原合成,并呈剂量依赖关系,其作用可能与下调HIF-1α、TGF-β1 mRNA表达有关。     

甲状腺素对血管紧张素Ⅱ所致心肌成纤维细胞胶原合成改变的影响

目的 :以AngⅡ处理培养的心肌成纤维细胞 ,促使细胞中胶原的合成发生改变 ,观测T3对AngⅡ所致细胞胶原合成改变的影响。方法 :以AngⅡ及T3分别或同时作用于培养的大鼠心肌成纤维细胞 ,通过测定3H TdR和3H Leu掺入来了解成纤维细胞增殖率和蛋白质合成率 ,同时用免疫细胞化学方法检测成纤维细胞中Ⅰ、Ⅲ型胶原的表达。结果 :AngⅡ使成纤维细胞3H TdR和3H Leu掺入增加 ,同时细胞中Ⅰ、Ⅲ型胶原的表达也显著增加。T3组成纤维细胞3H TdR和3H Leu掺入虽然也增加 ,但T3组细胞中Ⅰ、Ⅲ型胶原的表达却呈显著性降低。当T3与AngⅡ共同作用于成纤维细胞时 ,与T3组和AngⅡ组比较 ,3H TdR和3H Leu的掺入率没有发生显著性的变化 ;与AngⅡ组比较 ,细胞中Ⅰ、Ⅲ型胶原的表达显著性降低 ,与对照组的表达一致。结论 :T3能降低AngⅡ所致的心肌成纤维细胞中Ⅰ、Ⅲ型胶原合成的增加     

干扰素α对肝纤维化鼠星状细胞I和Ⅲ型前胶原mRNA表达及胶原？…

目的 观察干扰素α对大鼠纤维化时星状细胞增殖,Ｉ,Ⅲ型前胶原ｍＲＮＡ表达和肝脏胶原沉积的影响。方法 以ＣＣｌ４制造肝纤维化模型,培养肝星状细胞,抽提ＲＮＡ,用地高辛标记Ｉ,Ⅲ型前胶原和胶原酶ｃＤｎＡ探针,Ｎｏｒｔｈｅｒｎ杂交分析Ｉ,Ⅲ型前胶原和胶原酶ｍＲＮＡ表达,Ｄｏｔｂｌｏｔ测定大鼠肝Ｉ,Ⅲ型胶原沉积,分别用＾３Ｈ－ＴｄＲ和＾３Ｈ－脯氨酸掺入观察干扰素α对星状细胞增殖和胶原合成的影响。结果 干扰     

脂多糖对正常人皮肤成纤维细胞Ⅰ、Ⅲ型前胶原及胶原酶mRNA表达的调控及意义

目的研究细菌脂多糖（LPS）对增生性瘢痕患者正常皮肤成纤维细胞Ⅰ、Ⅲ型前胶原及胶原酶mRNA表达的影响,以了解LPS在增生性瘢痕形成中的生物学作用。方法体外培养增生性瘢痕患者瘢痕组织及正常皮肤成纤维细胞,应用不同浓度（0．005—1．0μg／m1）的大肠杆菌LPS（E．coli055：B5）对正常皮肤成纤维细胞进行刺激,并对刺激后细胞传代至表型稳定（第8代）,采用逆转录-聚合酶链反应（RT—PCR）法检测LPS对正常皮肤成纤维细胞Ⅰ、Ⅲ型前胶原及胶原酶mRNA的表达的调控作用,观察剂量一效应关系。分别以同一个体相同代数的瘢痕组织成纤维细胞和未经LPS刺激的正常皮肤成纤维细胞做阳性对照和阴性对照。结果LPS刺激浓度在0．005．0．5μg／ml范围内促进正常皮肤成纤维细胞Ⅰ、Ⅲ型前胶原mRNA表达,抑制胶原酶mRNA表达（均P〈0．01）,均在0．1μg／ml浓度点作用达高峰;当LPS刺激浓度到达1．0μg／ml时,抑制正常皮肤成纤维细胞Ⅰ、Ⅲ型前胶原mRNA表达,促进胶原酶mRNA表达,且均显著低于阴性对照组（均P〈0．01）。当LPS刺激浓度为0．1μg／ml时,正常皮肤成纤维细胞Ⅰ、Ⅲ型前胶原mRNA和胶原酶mRNA表达量与阳性对照组近似（均P〉0．05）。结论在一定浓度范围内LPS促进成纤维细胞Ⅰ、Ⅲ型前胶原mRNA表达、抑制胶原酶mRNA表达。LPS可能是增生性瘢痕形成的原始诱导因素之一。     

高氧对新生CLD鼠肺成纤维细胞增殖和Ⅰ型胶原合成的影响

目的 探讨持续吸入高氧后新生大鼠肺成纤维细胞的增殖情况及Ⅰ型胶原表达的动态变化.方法 足月新生鼠生后12h内分别持续吸入90%的高氧和空气,于3 d,7 d和14 d随机处死动物后,进行肺成纤维细胞的原代培养,应用流式细胞仪测细胞周期;免疫组化检测PCNA抗原;ELISA方法检测Ⅰ型胶原蛋白含量变化.结果 高氧组7 d和14 d肺组织成纤维细胞增生明显,S期比率显著高于空气组细胞,而G0/G1期比率显著低于空气组细胞(P<0.05),伴随Ⅰ型胶原蛋白表达的增加(P<0.05).结论 高氧可促进新生大鼠肺成纤维细胞过度增殖,Ⅰ型胶原合成增加,最终导致肺纤维化的发生.     

Value of D-Dimers in patients with acute aortic dissection

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.     

Research of combined liver-kidney transplantation model in rats

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.     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

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.     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

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.     

The comparative studies of the influences of Urapidil and Nicardipine on sino-atrial node function, atrio-ventricular node function and hemodynamics

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.     

Expression of OPGmRNA and ODFmRNA in the patients of hip fracture due to osteoporosis

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.     

The clinicopathological and immuohistochemical analysis of solid-pseudopapillary tumor of the pancreas： report of 9 cases

Objective：To investigate the clinical features, pathological characteristics and immunophenotype of solid-pseudopapillary tumor of the pancreas（SPTP）. Methods：Nine surgically treated cases of SPTP were retrospectively reviewed. Hematoxylin and Eosin（HE） staining and immunohistochemical staining were used to analyze all cases, and the general clinical data was collected. Results：Six patients were asymptomatic except for a palpable mass. Two patients complained of vague-epigastric pain. One patient appeared jaundice. The tumor was encapsulated and solid tissues alternately with cystic tissues. Histologically, the histological structure of solid portion was pseudopapillary with a fibrovascular core. Tumor cells were uniform and medium-sized which were arranged in sheets ets or nests or pseudopapillary patterns. Immunohistochemical studies demonstrated that SPTP proved positive in vimentin（9/9 cases）, AAT（9/9 cases）, NSE（9/9 cases）, ACT（7/9 cases）, CK20（2/9 cases）, CgA（1/9 cases）, S-100（3/gcases）, PR（4/gcases）, Syn（3/9 cases） and CD56（5/9cases）, negative in CEA and ER. Conclusion：SPTP is a tumor predominantly occurring in young women frequently without special symptoms. This tumor has various characteristical histological patterns with different immunophenotype.     

Dynamic Changes in Serum Estradiol and Progesterone levels in Patients of Premenstrual Syndrome with Adverse Flow of Liver-qi

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.     

Real-time Three-dimensional Echocardiography in Assessment of Left Ventricular and Right Ventricular Volumes

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.     

SCREENING FOR A 21-CHROMOSOME ABNORMALITY IN PREIMPLANTED EMBRYOS OF ELDERLY WOMEN

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.