佛波酯对体外培养大鼠前庭上皮PKC表达的影响

目的 观察体外培养条件下蛋白激酶C(protein kinase C,PKC)在佛波酯（phogrbol 12-myristate 13-acetate,PMA)诱导下大鼠前庭上皮的表达和分布特点。方法 取新生大鼠椭圆囊，建立体外前庭器官培养模型，采用免疫组织细胞化学的方法，以PKCβ，γ抗体为标记物，检测PMA诱导后前庭上皮PKCβ，γ的表达变化和分布特点。结果 PKCβ，γ在正常培养新生大鼠前庭上皮细胞的细胞膜及细胞质中有弱表达。PMA作用后，其表达逐渐增多，PKCβ1主要位于细胞膜，在30min时表达最强，其后逐渐减少，45min仍高于正常（P＜0．01）。PKCβ2不但位于细胞膜，而且在纤毛亦明显表达；PKCγ在PMA诱导后胞质表达增加。在毛细胞层和支持细胞层均有，主要位于毛细胞层。结论 体外前庭器官培养在PMA诱导后，PKC各亚型激活的形式及程度不同。     

庆大霉素对豚鼠前庭上皮IGF-1及其受体表达的影响

目的：研究庆大霉素对豚鼠前庭上皮胰岛素样生长因子-1（insulin-like growth factor-1，IGF-1）及其受体（insulin-like growth factor-1 receptor，EGF-1R）表达的影响，并探讨其生物学特性。方法：采用免疫组织细胞化学方法，以抗IGF-1及IGF-1R抗体为标记物，检测庆大霉素损伤后豚鼠前庭上皮自发修复期IGF-1及IGF-1R的表达变化和分布特点；采用Brdu体内标记和抗Brdu抗体免疫组化染色，检测庆大霉素损伤后豚鼠前庭上皮增殖的变化。结果：IGF-1及IGF-1R在正常成年豚鼠前庭上皮细胞中有低水平表达，庆大霉素损伤后，其表达增多。在1d组IGF-1及IGF-1R表达最强，其后逐渐减少。在正常对照组中无Brdu阳性细胞。各实验组均观察到Brdu阳性细胞，7d组最多。结论：庆大霉素损伤后，豚鼠前庭上皮IGF-1及IGF-1R表达增强，其时程变化与细胞增殖活动密切相关。IGF-1在豚鼠前庭上皮损伤后的细胞增殖中可能起重要信号转导作用。     

碱性成纤维细胞生长因子及成纤维细胞生长因子受体在豚鼠前庭中的定位和表达

目的 :观察碱性成纤维细胞生长因子 ( b FGF)及成纤维细胞生长因子受体 ( FGFR)在豚鼠前庭终器的定位和表达。方法 :免疫组织化学。结果 :b FGF位于支持细胞和毛细胞的胞浆及胞核 ,FGFR位于支持细胞表面。b FGF及 FGFR在成年豚鼠表达较弱 ,新生豚鼠及庆大霉素内耳损伤豚鼠表达较强。结论 :提示 b FGF在豚鼠前庭终器发育、结构功能维持及损害后修复中起重要作用     

胃癌耐药细胞系传统型蛋白激酶C的表达及活性

目的 观察传统型蛋白激酶C（cPKCs）在胃癌耐药细胞系SGC7901／VCR1.0、耐药亚系SGC7901／VCR0.3,7901/0.7及药敏细胞系SGC7901表达及活性的变化,以及对细胞内药物浓度的影响。方法 用免疫荧光化学及蛋白印迹方法观察传统型蛋白激酶C在胃癌耐药细胞系及药敏细胞系的表达;竞争蛋白结合法测定PKC的活性;应用流式细胞仪观察细胞内药物浓度的变化。结果 PKCα,PKCβⅠ,PKCβⅡ及PKCγ在胃癌耐药细胞系及药敏细胞系均有表达,PKCα在耐药细胞表达呈强阳性,在药敏细胞表达呈阳性;PCKβⅠ及PKCβⅡ在耐药细胞及药敏细胞表达均为阳性;PKCγ在耐药细胞及药敏细胞表达均为强阳性。免疫蛋白印迹实验证实,随耐药指数的增加,PKCα表达呈逐渐增高的趋势,薄层扫描蛋白印迹带的吸光度（A）分别为9584.17,9421.06,8534.64,8088.79,而PKCβⅠ、PKCβⅡ及PKCγ在耐药细胞系、耐药亚系及其药敏细胞系表达无明显变化。PKC活性检测结果提示,随耐药指数的增加,PKC活性呈逐渐增高的趋势,以细胞质及细胞核的PKC活性增高为主。结论 传统型蛋白激酶C在维持胃癌耐药细胞系SGC7901／VCR的多药耐药中起重要作用且具有同工酶特异性。     

碱性成纤维细胞生长因子及成纤维细胞生长因子受体在豚鼠前 …

目的：观察碱性成纤维细胞生长因子（ｂＦＧＦ）及成纤维细胞生长因子受体（ＦＧＦＲ）在豚鼠前庭终器的定位和表达。方法：免疫组织化学。结果：ｂＦＧＦ位于支持细胞和毛细胞的胞浆及胞核，ＦＧＦＲ位于支持细胞表面。ｂＦＧＦ及ＦＧＦＲ在成年豚鼠表达较弱，新生豚鼠及庆大霉素内耳损伤豚鼠表达较强。结论：提示ｂＦＧＦ在豚鼠前庭终器发育、结构功能维持及损害及修复中起重要作用。     

豚鼠前庭上皮扫描电镜观察

研究豚鼠前庭上皮的扫描电镜结构。方法：制作 豚鼠前庭扫描电镜标本,用Ｓ－５２０型扫描电镜观察。结果：前庭上皮由Ⅰ型、Ⅱ型毛细及支志细胞表有呈阶梯状排列的静纤毛及一根动纤毛,两种毛细胞大小,纤毛排列方式均不同相同。结论：正常豚鼠前庭毛细胞分为两型Ⅰ型毛细胞大,其静纤毛长,Ⅱ型毛细胞相对小,其静纤毛短。     

豚鼠发育过程前庭上皮ERK 1/2的表达

目的：检测胞外信号调控蛋白激酶1/2（extracellu-lar signal regulated kinase，ERK1/2）在豚鼠发育过程前庭上皮的表达和分布特点，并探讨其生物学特征及意义。方法：利用免疫组织化学的方法，以ERK1，ERK2抗体为标记物，检测发育期豚鼠前庭器中的表达情况及分布特点；利用West-ern bolt方法检测ERK1/2的活性变化。结果：ERK1/2可在豚鼠前庭上皮细胞中表达，随年龄增加而逐渐减少。ERK1/2的活性随年龄的增长而逐渐降低，且ERK2的活性高于ERK1。结论：正常豚鼠前庭上皮中有ERK1/2表达，其表达量及活性与细胞增殖、分化相一致，随发育过程而减少。     

肿胀激活状态下蛋白激酶C同工酶亚型在胃癌耐药细胞系的亚细胞分布变化及其意义

目的 探讨肿胀激活状态下，传统型蛋白激酶C（cPKC)同工酶亚型在胃癌SGC7901及其耐药细胞系SGC7901/VCR的表达，亚细胞分布变化及其意义。方法 采用免疫荧光及免疫印迹方法，观察正常状态及持续低渗灌流不同时间cPKC同工酶亚型的亚细胞分布变化，利用免疫荧光双标记及共聚焦显微镜技术观察PKCα与-P-糖蛋白（Pgp)的共表达。结果 在正常状态下，cPKC的4种同工酶亚型在胃癌SGC7901及其耐药细胞SCG7901/VCR细胞质及细胞膜均有表达：PKCα在耐药细胞表达呈强阳恬药敏细胞表达呈强阳性，PKCγ在耐药细胞及药敏细胞表达均呈强阳性。持续低渗灌流，在耐药系10min已发现有PKCα、PKCγ的移位及细胞体积的增大；30minPKCα几乎全部移位至细胞膜上，PKCγ部分移位至细胞核内，细胞体积较前增大1-2倍；60min时PKCα及PKCγ基本恢复至正常位置。细胞体积也恢复正常；在药敏细胞系10-20minPKCα及PKCγ几乎全部一笔勾销 位至细胞膜上，PKCγ部分移位至细胞核内，细胞体积也恢复正常；在药敏细胞系10-20minPKCα几乎全部移位至细胞膜上，PKCγ部分移位至细胞核内，细胞体积较前增大1-2倍；40minPKCα及PKCγ基本恢复至正常位置，细胞体积也恢复正常，而PKCβ1及PKCβα在耐药细胞及药敏细胞的亚细胞分布无明显变化。共聚焦显微镜提示PKCα及PKCγ同工酶亚型参与了Pgp与低渗刺激下细胞共表达，以耐药细胞表达明显。结论 只有PKCα及Pgp表达量有关，而PKCβ1及PKCβⅡ可能与这一过程无关。βββ     

解剖、生理和组胚学

021,27胚胎期豚鼠前庭终器细胞凋亡的检测/胡金玮…//第四军医大学学报一2002,23(16)一1514一1517 研究豚鼠胚胎正常发育期前庭终器形态变化及细胞凋亡,探讨细胞凋亡在前庭终器发育中的作用。 ·328.方法:选用胚胎40d和50d正常豚鼠,光镜观察其前庭终器发育形态变化,末端脱氧核昔酸转移酶介导的生物dUTP缺口末端标记技术(TUNEL)测定毛细胞和支持细胞凋亡,并以电镜观察凋亡细胞核变化。结果:胚胎40d,豚鼠前庭壶腹、椭圆囊斑及球囊斑已分化,前庭上皮出现一层毛细胞及多层支持细胞;胚胎50d,支持细胞层数减少,毛细胞数目增加,伴有神经纤维出…     

蛋白激酶CβⅡ负调控小鼠骨髓间充质干细胞的成骨分化

目的观察蛋白激酶C(PKC)各亚型在小鼠骨髓间充质干细胞(BMSCs)成骨分化中的作用。方法Western blot检测小鼠BMSCs成骨分化中PKC各亚型的活化情况;利用PKC阻断剂GFX分析PKC在BMSCs体外成骨分化中的作用;碱性磷酸酶(ALP)试剂盒检测成骨早期分化指标ALP的活性;茜素红染色检测BMSCs体外矿化能力;实时荧光定量聚合酶链反应(PCR)检测成骨分化相关基因Runx2、Ⅰ型骨胶原a1(ColⅠa1)和ALP的表达情况。结果小鼠BMSCs成骨分化过程中,仅检测到PKCβⅡ亚型活化,其他亚型均无明显活化。利用PKC阻断剂GFX抑制PKCβⅡ活化后,茜素红染色结果显示BMSCs的成骨分化过程受到促进;PCR结果显示成骨分化相关基因Runx2、ColⅠa1和ALP的mRNA水平均有不同程度增加;ALP活性检测结果也显示BMSCs的成骨分化过程受到促进。结论PKCβⅡ可负调控小鼠BMSCs的成骨分化。     

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.