自然流产患者滋养细胞肝素表皮生长因子的表达及意义

目的　探讨自然流产患者滋养细胞肝素表皮生长因子 (HB EGF)的表达及其与滋养细胞增殖细胞核抗原 (PCNA)的关系。方法　选择自然流产患者和正常妊娠者各 2 0例 ,采用免疫组织化学法分析滋养细胞HB EGF和PCNA的表达。结果　HB EGF主要存在于细胞质 ,合体滋养层和细胞滋养层均有表达 ,部分流产滋养细胞不表达。自然流产患者滋养细胞HB EGF和PCNA的表达显著低于正常妊娠者 (P <0 .0 1) ,成正相关 (r =0 .4 5 4 ,P <0 .0 1)。结论　HB EGF的表达和滋养细胞增殖相关 ,HB EGF下降可能与自然流产的发生相关     

在体诱导表皮细胞去分化的初步实验研究

目的 在体诱导表皮细胞去分化,探索与表皮细胞去分化有关的信号通路.方法 包皮皮片去除皮下组织,用中性蛋白酶分离表皮,Ⅳ型胶原反复粘贴并牵拉表皮去除基底细胞层.处理后表皮片移植于裸鼠全层皮肤下,分别于3、5、7 d取出移植皮片,采用免疫组织化学染色、流式细胞仪检测、Western blot和RT-PCR等方法检测皮片移植前后表型的改变和ERK MAPK通路各信号分子表达的变化情况.结果 去基底细胞层处理的表皮片干细胞标志物CK19和β1整合素染色阴性;移植后5 d,存活皮片CK19和β1整合素染色呈多层分布,而不是正常表皮中的单层分布.流式细胞仪检测结果显示,表皮片移植后CK19阳性细胞和β1整合素阳性细胞由移植前的0.00%分别增加到7.54%和5.24%,而CK10阳性细胞由99.62%下降为86.56%.Western blot和PCR检测也证实CK19和β1整合素蛋白水平和mRNA水平表达在移植后明显增强.ERK及上下游信号分子(p-Raf、p-MEK1/2、p-ERK1/2和c-myc)表达增强.t-MEK1/2和t-ERK1/2移植前后表达无显著差异(P＞0.05).结论 处于终末分化阶段的表皮细胞可以向干细胞状态逆转,这个过程可能与ERK MAPK信号通路有关.     

转染hEGF基因的表皮细胞G418长期筛选后的表达

目的:探讨外源性基因在转染表皮细胞后稳定表达情况.方法:选用构建的pBK-Signal-EGF质粒,在脂质体LipofectAMINE的介导下,转染培养成60%～70%连片的人表皮细胞,然后用G418长时间筛选出稳定表达的细胞克隆.大量培养传代后,通过ELISA试剂盒检测培养上清中表皮细胞生长因子(EGF)外源性基因稳定表达水平.结果:(1)构建的人表皮细胞生长因子(hEGF)质粒可成功转染到人表皮细胞中;(2)转基因表达在传4代后仍可存在;(3)hEGF表达水平6周内可在培养基中测出.结论:外源性hEGF基因可以在表皮细胞中稳定表达.     

整合素β1在人表皮细胞株HaCaT增殖中影响的研究

目的探讨β1整合素对表皮细胞增殖的影响.方法将重组整合素β1小干扰RNA表达载体(siIntegrin β1)转染到HaCaT细胞株中,应用免疫印迹(Western blotting)方法检测靶基因的抑制情况,应用细胞计数法检测表皮细胞增殖情况.结果重组siIntegrinβ1 转染HaCaT细胞株能够抑制靶基因整合素β1 的表达,抑制细胞增殖.结论在人表皮细胞株HaCaT中抑制整合素β1表达可抑制HaCaT增殖,为进一步研究整合素在表皮细胞中的基因表达调控奠定了基础.     

一种稳定、有效的表皮细胞基因导入方法的确立

目的：探索一种表皮细胞基因转染的稳定有效方法。方法：我们用五种方法将报告基因ｐＳＶ－β转染培养的表皮细胞,用原位染色方法检测并结合图像分析比较这五种方法的转染后表达情况。结果 转染后２４ｈ表皮细胞开始表达报告基因,７２ｈ达到高峰,脂质体结合１０％甘油休克法的转染效率达到４．２％。结论 表皮细胞难于转染,脂质体结合１０％甘油休克法是一种稳定、有效的转染方法。     

Mfn2基因过表达对人乳腺癌MCF-7细胞增殖及EGFR、EGF蛋白表达的影响

目的 通过使人乳腺癌MCF-7细胞过表达线粒体融合素基因-2(Mfn2),研究外源性Mfn2基因对MCF-7细胞增殖及对表皮生长因子受体(EGFR)、表皮生长因子(EGF)蛋白表达的影响.方法 实验分为对照组、转染空质粒pEGFP-N1组、转染Mfn2质粒的pEGFP-Mfn2组.转染48 h后,检测各组细胞的转染效率、Mfn2mRNA及Mfn2蛋白表达.检测各组MCF-7细胞增殖情况、各组MCF-7细胞周期分布情况.同时检测各组EGFR及EGF蛋白表达情况.结果 转染48 h后pEGFP-N1组及pEGFP-Mfn2组转染效率分别为(49.15±2.04)％及(51.10±2.18)％,高于对照组[(0.58±0.21)％,P＜0.05];pEGFP-Mfn2组的Mfn2 mRNA及Mfn2蛋白表达均显著高于对照组及pEGFP-N1组(P＜0.05);pEGFP-Mfn2组MCF-7细胞增殖抑制率显著高于其余两组(P＜0.05),细胞周期主要阻滞于G0/G1期;pEGFP-Mfn2组EGFR和EGF蛋白表达水平显著低于其余两组(P＜0.05).结论 Mfn2基因过表达可显著抑制MCF-7细胞增殖,其机制可能与Mfn2基因过表达导致了EGFR及EGF表达下调有关.     

~(60)Co照射后小鼠表皮Langerhans细胞的变化及对异体植皮的影响

本实验用~(60)Co照射Blab/c小鼠,照射后1天与3天取背部皮肤。(1)经ATP酶染色方法,显示表皮Langerhans细胞(LC),观察其ATP酶含量的变化、形态变化及其在表皮的密度改变;(2)免疫组分染色方法,观察Ia阳性LC的密度改变及形态变化;(3)电镜观察其超微结构的变化;(4)同时进行异体皮肤移植,观察皮片存活期的变化。结果:经~(60)Co照射后,小鼠表皮LC形态功能受到不同程度的损害,其密度减少,表达Ia抗原功能下降。认为这是异体植皮后移植物存活期延长的主要原因。     

α干扰素对尖锐湿疣患者皮损表皮细胞凋亡和增殖的影响

目的　研究α -干扰素对尖锐湿疣患者皮损表皮细胞凋亡和增殖的影响。方法　采用免疫组织化学方法和末端脱氧核苷酸转移酶介导的末端标记技术 (TUNEL法 ) ,检测 40例患者皮损表皮细胞凋亡和增殖状态 ,并对细胞凋亡和PCNA阳性细胞进行计数。结果　α 干扰素治疗后PCNA阳性细胞数明显减少而凋亡细胞数增加 ,与治疗前比较差异均有显著性 (P <0 .0 5)。结论　INF α能促进细胞凋亡及抑制细胞增殖     

庆大霉素中毒后雏鸡耳蜗毛细胞再生和表皮生长因子的促再生作用

目的:观察雏鸡庆大霉素(GM)中毒后耳蜗毛细胞的再生及表皮生长因子(EGF)对其再生的影响.方法:给雏鸡注射GM10d,其后再注射EGF5d,分别用光镜、扫描电镜及透射电镜观察耳蜗基底乳头(BP)形态学变化;用增殖细胞核抗原(PCNA)免疫组织化学染色观察BP细胞增殖.结果:停用GM后1d,BP毛细胞损失平均约51%,毛细胞形态不规则,线粒体和表皮板肿胀,细胞自溶破坏.停用GM后5d,支持细胞表面增宽,中间出现小的表皮板和短的听毛,此为新的毛细胞.14d新生毛细胞形态规则,21d发育成熟,BP形态接近正常.PCNA阳性细胞主要是支持细胞和透明细胞.EGF组毛细胞再生活跃,PCNA阳性细胞核明显增多.停用GM后10d,毛细胞数明显比GM组多,14dBP形态接近正常.结论:GM耳中毒的雏鸡耳蜗毛细胞可以再生;再生的毛细胞可能来源于支持细胞和透明细胞;EGF可促进这一过程.     

人表皮细胞在牛胶原膜粘附、生长及分化的研究

目的在牛胶原膜中植入胎儿表皮细胞和成纤维细胞，构建人工复合皮替代物，以研究细胞的粘附、生长、分化及真表皮间的相互作用。方法(1)酶消化法培养胎儿表皮细胞和成纤维细胞；(2)制备人工复合皮，接种表皮细胞后于不同时间通过HE染色和扫描电镜观察细胞的形态和数量变化；(3)透射电镜观察该复合皮中细胞的结构；(4)裸鼠移植试验验证该复合皮的作用。结果(1)该人工复合皮具有一定的弹性，可出现较明显的细胞增殖，未见人工复合皮收缩现象。(2)HE染色和扫描电镜下可见随培养时间的延长，成纤维细胞和表皮细胞逐渐增多，表皮细胞可发生分化。并逐渐分层。(3)透射电镜下见成纤维细胞和表皮细胞功能活跃。(4)裸鼠移植试验证实该人工皮可起到覆盖创面防止感染的作用。结论该人工复合皮基本具备了正常皮肤的结构和功能，可作为移植物覆盖全层皮肤损伤。     

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.