甲胎蛋白在不同发育时期大鼠胃组织中的表达及定位

目的：研究大鼠正常发育不同阶段胃组织中甲胎蛋白（AFP）的表达及定位.方法：取SD大鼠发育18.5,20.5d胚胎及出生0,7,14,21d和8wk大鼠的胃组织.采用RT-PCR,WesternBlot方法检测AFP的表达;采用免疫荧光染色技术检测AFP的表达及细胞定位.结果：在18.5d胚胎大鼠胃组织中,AFPmRNA表达量显著高于其它各时期,出生后AFP蛋白表达量稳步下降,20.5d胚胎大鼠及出生后0,7,14,21d大鼠AFPmRNA表达量分别是18.5d胚胎大鼠表达量的85%,58%,37%,24%,11%,差异均具有统计学意义（P〈0.05）,成年后消失.AFP蛋白表达量变化趋势与mRNA表达量变化趋势基本一致.免疫荧光染色的结果显示：AFP在大鼠18.5d胚胎至出生后21d期间各时期大鼠胃组织中与间充质细胞标志物波形蛋白（Vimentin）呈共表达.结论：AFP在胚胎发育晚期的高表达以及在发育各个时期均特异性表达于大鼠胃间充质细胞,提示AFP可能对胃结构的形成和功能的完善起重要作用.     

大鼠睾丸间质细胞发育的形态学研究

采用光、电镜技术对６３例胚胎及出生后不同发育时期大鼠睾丸间细胞的发育进行了研究。结果表明：（１）大鼠睾丸间质细胞自胚胎１５天出现，胚胎１７天已初具分泌类固醇激素的结构特点；（２）大鼠睾丸间持细胞幼稚型、成熟型和退化型３种形式，但其分化、成熟与退化是相伴存在的；（３）大鼠丸间质细胞在整个发育过程中存在２个细胞增殖峰；胚胎１７－１９天和青春期。二峰之间有一个低谷，在约在生后第２周；（４）生后第２周后，     

大鼠胰腺不同发育时期转录因子Nkx2.2和Nkx6.1 mRNA的表达调控研究

目的：研究大鼠胚胎胰腺发育后期、新生期及成年期对转录因子Nkx2．2和Nkx6．1的mRNA表达调控。方法：利用显微分离等方法分离胚胎18．5天、新生和成年大鼠胰腺,对胰腺标本提取总RNA．利用胰腺内外分泌细胞分子标志——胰岛素和淀粉酶来鉴定胰腺标小,再用RT-PCR分析Nkx2．2或Nkx6．1存大鼠胰腺几个不同发育时期的mRNA表达情况结果：Nkx6．1在胚胎18．5天胰腺中有表达,出生时表达量有增加,到了成年显著下降。Nkx2．2的mRNA存胚胎期、新生期和成年期均有表达,新生期表达量较其他两个时期有显著性增加。从以上两个转录因子在胰腺不同发育时期的表达情况来看．它们的其同点是胚胎期有表达,新生期表达量较胚胎后期和成年期高,成年期表达量呈下降趋势。结论：不同生长发育时期对转录因子Nkx2．2和Nkx6．1的mRNA具有调节作用,新生期细胞增生分化加速,这与Nkx2．2、Nkx6．1等转录因子表达量增高有关。     

大鼠气管发生的形态学观察

本实验对胚胎第15天至出生后30天的大鼠气管上皮的发生过程进行了观察。证实胚胎第15天的气管上皮为假复层上皮,细胞表面有少量微绒毛;胚胎第17天,气管上皮出现典型的杯状细胞;胚胎第19天出现纤毛细胞;出生5天以后,上皮细胞增殖、聚集并内陷,形成气管腺。杯状细胞的粘液颗粒主要是中性粘多糖,气管腺的粘液颗粒为酸性粘多糖和中性粘多糖。实验结果表明:在胚胎期和生后早期,粘液纤毛运送系主要由纤毛细胞和杯状细胞等组成,出生5天以后,粘液纤毛运送系统主要由纤毛细胞和气管腺组成。     

BMP4mRNA在Meckel''''s 软骨中的表达及意义

目的：观察和分析BMP4mRNA在Meckel’s软骨细胞中的表达，探讨其在下颌骨发育中的调控作用．方法：建立和制备sD大鼠胚胎时期13～21d等多时间点的下颌骨发育模型，采用原位杂交检测：BMP4mRNA在Meckel‘s软骨中的表达．结果：BMP4mRNA在胚胎SD大鼠下颌骨发育的各个时期中呈一定的时空表达，在E13～17d的Meckel‘s软骨细胞中呈阳性表达；E19d时呈强阳性表达；E21d时呈弱阳性表达．结论：BMP4mRNA在Meckel‘s软骨细胞中的表达特点，提示Meckel‘s软骨细胞中的BMP4mRNA与诱导间充质细胞分化及下颌骨形成有关．     

HNF4α和HNF6 mRNA在小鼠肝发育过程中的表达

目的 探讨HNF4α和HNF6在肝发育过程中的作用。方法 用RT-PCR和原位杂交检测HNF4α mRNA和HNF6 mRNA在胚胎第8、9、13、15、17d(E8、E9、E13、E15、E17)、出生后第1d(P1)以及成年小鼠肝中的表达。结果 RT-PCR结果显示，HNF4α在肝中的表达始于E9，并持续到新生，至成年时表达减弱。原位杂交结果显示．在胚胎发育的各个阶段多数肝索细胞呈HNF4α阳性反应，成年时仍有少数HNF4α阳性的肝索细胞，而胆管板及胆管上皮细胞、血管内皮细胞、造血细胞等均为阴性。HNF6 mRNA的表达在E9开始出现，E13消失，E15又重新出现，并一直维持到出生。E9、E15多数肝索细胞呈HNF6 mRNA阳性反应，E17及P1 HNF6 mRNA在肝索细胞中的反应减弱，胆管板细胞出现强阳性反应。结论 HNF4α mRNA表达与肝芽的形成有关，并参与肝干细胞向肝细胞的增殖、分化的过程，其在正常成年肝中的表达可能与肝细胞正常形态的维持有关。HNF6可能在肝干细胞的形成和肝干细胞向胆管细胞的分化，以及胆管细胞的增殖、分化过程中发挥作用，其在正常成年肝中可能介导维持胆管上皮细胞的正常形态。     

脱氢表雄酮在大鼠胃上皮细胞中的免疫定位

目的：研究脱氢表雄酮（DHEA）在大鼠胃上皮细胞中的定位。方法：采用免疫组织化学SABC法。结果：大鼠胃小凹上皮细胞、胃底腺壁细胞呈DHEA免疫反应阳性,阳性物质分布于胞质内,胞核阴性。胃底腺其他细胞呈免疫反应阴性。论：以上结果提示,胃小凹上皮细胞,胃底腺壁细胞含有DHEA,DHEA对消化道可能具有重要的调节功能。     

C57小鼠出生后视网膜神经上皮层细胞发育中的PCNA表达及其意义

目的　研究C5 7小鼠出生后视网膜的发育和增殖细胞核抗原 (PCNA)表达及两者间的关系。方法　采用HE染色及免疫组化标记PCNA的方法 ,对出生后第 1天到第 2 1天的C5 7小鼠视网膜神经层的细胞发育作连续性观察。结果　C5 7小鼠出生后 ,视网膜神经层仍然有一个发育过程。神经节细胞层在出生时已经形成 ,外丛状层在出生后第 6天才分化出来 ,而内、外核层细胞形态上接近成熟小鼠 ,分别在出生后第 10天、19天。PCNA显示在外成神经细胞层分化为内、外核层和外丛状层之前 ,和节细胞各有一次和增殖相关的阳性表达。 3个细胞层在形态学的分化基本完成后 ,又出现了一次结束时间是由外向内变化的阳性过程 (外核层 :第 11天至 15天 :内核层 :第 8天至 16天 ,神经节细胞层 :16天到 2 1天 ) ,这一次阳性表达过程可见到细胞形态向成年形态的转化。结论　C5 7小鼠出生后视网膜神经层的发育 ,分为细胞分化和细胞成熟两个阶段 ,分化阶段的细胞分化顺序可能是以细胞的空间位置决定的 ,成熟阶段的顺序可能是按神经传导的方向发展的 ,与细胞的功能相关 ,PCNA的表达和细胞的增殖、分化、成熟在时间、空间上具有相关性。     

pax2基因在肾小管上皮细胞转分化中的作用

目的 探讨pax2基因在大鼠慢性肾小管间质纤维化过程中的表达及其对肾小管上皮细胞转分化的可能作用.方法 大鼠5/6肾切除造模,分别于术后2、4、6、8、10、12周取左肾皮质,采用逆转录(RT)-PCR及免疫组织化学的方法检测模型组和正常对照组的pax2和α-SMA的表达.结果 病变组织见系膜细胞和基质增生,间质炎症细胞浸润,基底膜变厚.5/6肾切除模型组大鼠在术后第2周出现pax2 mRNA的强表达,第4周达高峰,与正常对照组比较P＜0.05,此后下降至痕量表达;α-SMA mRNA也在术后第2周表达逐渐增强,第8周达高峰,与正常对照组比较P＜0.05,此后逐渐下降.免疫组织化学染色显示,5/6肾切除模型组pax2蛋白的表达、肾小管上皮细胞中α-SMA的表达与pax2 mRNA、α-SMA mRNA的表达趋势相同.结论 在大鼠慢性肾间质纤维化的过程中,受损的肾小管上皮细胞可向肌成纤维细胞转分化,肾小管发育胚胎基因pax2在此过程中的一过性表达可能起到重要的作用.     

孕大鼠在胚泡着床前给予醋氨酚对器官形成期胚胎发育的影响

研究孕大鼠胚泡着床前药物损伤对胚胎器官形成期发育的影响，探索药源性胚泡异常与胎仔发育的关系。方法：母鼠孕第3天给予醋氨酚0．25～1．00g／kg．孕第4天处死母鼠．计数胚泡细胞数，胚泡微核数及微核率；孕第12．5天处死母鼠，评价胚胎组织器官形态分化程度及胚胎生长发育状况。结果：醋氨酚（0．25～1．00g／kg）抑制着床前胚泡发育,对胚泡细胞产生遗传毒作用，并呈明显剂量依赖关系。与对照组相比，孕第12．5天（器官形成期）胚胎生长发育状况无明显差异，胚胎组织器官形态分化无显著改变。醋氨酚0．5g，1．0g／kg对胚胎发育具毒性，导致胚胎发育迟缓、胚胎畸形、胚胎死亡数显著增加。结论：胚胎发育毒性与着床前胚泡遗传物质损伤相关。     

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.