DMBA/巴豆油二阶法联合NB-UVB照射构建小鼠皮肤鳞癌模型

目的用二甲基苯并蒽(DMBA)/巴豆油二阶法联合NB-UVB照射,构建皮肤鳞状细胞癌小鼠模型。方法将50只BALB/c小鼠,随机分为3组:A组外涂DMBA/巴豆油;B组照射NB-UVB;C组外涂DMBA/巴豆油,联合NB-UVB照射。实验过程中定期观察小鼠的一般情况和实验部位皮肤的变化,于第5、10、15、20周统计各组小鼠的存活率及成瘤率。第20周时取小鼠皮损组织进行病理检查。结果第5周末,部分C组小鼠出现直径≥1 mm的丘疹,第20周后,三组成瘤率分别为86.67%,7.14%,94.12%;SCC发生率分别为:13.34%,0%,70.59%。结论 DMBA/巴豆油二阶法联合NB-UVB构建小鼠皮肤鳞癌模型较单独应用DMBA/巴豆油二阶法或NB-UVB照射成瘤时间短,成瘤率和SCC发生率高。     

UVC因素对DMBA诱导的小鼠皮肤肿瘤建模的影响

目的观察短波紫外线(UVC)照射对7,12-二甲基苯蒽(DMBA)诱导的皮肤肿瘤建模的效应。方法采用完全随机的两因素析因设计。选择对光线敏感的BALB/c小鼠,背部皮肤脱毛后UVC照射(56 mJ/cm2),隔天1次,8周结束;DMBA/丙酮液(100μg/200μL)外涂,每周1次,共7次,12周后处死。连续测量并记录小鼠背部肿瘤数量和直径,描绘时间-荷瘤数动态变化图;皮肤组织病理学观察肿瘤形成及光损伤后的皮肤改变,并用计算机图像分析系统定量分析。结果单纯UVC照射组无肿瘤出现;UVC联合DMBA模型组小鼠6周后背部开始出现典型的上皮乳头状瘤,第9周荷瘤率达到100%,至11、12周荷瘤数渐趋稳定,平均荷瘤数为(4.57±3.0)个,肿瘤平均体积为(44.91±4.6)mm3。而单纯DMBA组荷瘤数第8.5周达高峰,后呈下降趋势,肿瘤自然消退率高,数量不稳定。结论 DMBA是皮肤肿瘤建模的主要作用因素,UVC作为一个独立因素,在12周内未能诱导出皮肤肿瘤,但UVC与DMBA联合应用具有交互协同作用,可较快速地建立荷瘤率高、瘤体数量稳定的皮肤肿瘤模型,较单纯DMBA建模效果好。     

皮肤光损伤肿瘤模型的建立

目的模拟人皮肤肿瘤形成的自然环境因素,建立光损伤小鼠皮肤肿瘤模型。方法采用完全随机的两因素析因设计。选择对光线敏感的BALB/c小鼠,背部皮肤脱毛后UVC照射(56 mJ/cm2),隔天1次,8周结束;二甲基苯蒽/丙酮液(100μg/200μL)外涂,每周1次,共7周,12周后处死。连续测量并记录小鼠背部肿瘤数量和直径,描绘时间-荷瘤数动态变化图;皮肤组织病理学观察肿瘤形成的组织细胞形态学改变。结果单纯UVC照射组无肿瘤出现;UVC+DMBA模型组小鼠6周后背部开始出现肉眼可见的瘤体,第9周荷瘤率达到100%,11、12周荷瘤数渐趋稳定,肿瘤体积有增大趋势,平均荷瘤数为(4.57±3.0)个,肿瘤平均体积为(44.91±4.6)mm3。HE染色、瑞氏染色、基底膜带染色均显示为早期皮肤鳞状细胞癌。而单纯DMBA组荷瘤数第8.5周达高峰,后呈下降趋势,肿瘤自然消退率高,数量不稳定。结论 DMBA是皮肤肿瘤建模的主要因素,但停止该处理因素后荷瘤数量呈下降趋势;UVC作为一个独立因素,在12周内未能诱导出皮肤肿瘤,但UVC与DMBA联合应用具有交互协同作用,可较快速地建立皮肤肿瘤模型。该模型具有出瘤率高,出瘤整齐,荷瘤量多,除去处理因素后瘤体数量依旧稳定,且体积有增大趋势等特点,为皮肤肿瘤的防治研究提供有价值的实验工具。     

高剂量7,12-二甲基苯并[a]蒽诱发的小鼠皮肤鳞状细胞癌模型建立及评价

目的:建立使用高剂量7,12-二甲基苯并[a]蒽诱发的小鼠皮肤鳞状细胞癌(SCC)模型,为皮肤鳞状细胞癌的预防与治疗提供实验基础。方法:将36只BALB/c小鼠随机分为3组,分别为:高剂量组(A组)、低剂量组(B组)、空白对照组(C组),造模过程中记录小鼠体重及背部皮损变化,造模后取局部皮肤组织进行HE染色及PCNA免疫组织化学染色,以PCNA阳性染色的表达水平作为反映小鼠皮肤鳞状细胞癌的恶性指标,使用Image-Pro Plus计算PCNA染色的积分光密度(IOD)和阳性面积,并对结果进行统计学分析。结果:12周时A、B两组的荷瘤数目、肿瘤体积比较有统计学差异(P<0.05),且荷瘤率分别可达83.3%、66.7%,SCC发生率分别可达75%、33.3%;免疫组织化学分析结果显示3组的IOD及阳性面积比较,差异有统计学意义(P<0.01)。结论:相较于传统二阶法建立的小鼠皮肤鳞状细胞癌模型,本模型具有实验周期更短、成瘤率更高、操作更为简便的特点,为皮肤鳞状细胞癌的研究提供了可靠的实验基础。     

二甲基苯蒽/佛波酯两步化学法诱导小鼠皮肤癌的模型优化

目的 对两步化学诱导法建立BALB/c小鼠皮肤癌模型进行方法优化。方法 将BALB/c小鼠背部剃毛暴露出约2 cm×2 cm的皮肤,小鼠随机分为空白对照组和4个模型组,4个模型组第1周分别涂1、2、4、7次100 μg/100 μL DMBA/丙酮液,后续均1周涂两次4 μg/100 μL TPA/丙酮液,记录小鼠体重、成瘤时间、成瘤数量,并在12周后处死小鼠取肿瘤组织及瘤旁组织做HE染色分析。结果 1周涂1次DMBA成瘤周期长;1周两次成瘤周期短,质量高;1周4次会损伤皮肤但不影响肿瘤的形成;1周7次对皮肤伤害较大并导致小鼠死亡。结论 采用第1周涂两次DMBA启动,后续用TPA连续诱导建立BALB/c小鼠皮肤癌模型操作简便,成瘤期短,癌变率高,与人类皮肤癌发生发展相似,可用于建立研究皮肤癌较为理想的实验动物模型。     

人结肠癌动物皮下实体瘤模型的建立

 摘 要：【目的】 建立临床背景清楚的结肠癌动物皮下实体瘤模型,为结肠癌发病机制及治疗策略的研究提供可靠的实验动物模型。【方法】 采用组织块直接移植法将收集到的12例结肠癌组织临床标本移植到免疫缺陷小鼠右侧肩胛皮下,第1-2代用NOD-Scid小鼠;将每个病例的第二代皮下肿瘤,分别连续皮下接种至NOD-Scid小鼠和BALB/C裸小鼠两组动物中,传代至第5代。观察每个病例在两种品系动物的成瘤时间、标本的成瘤率;进行HE染色和CDX-2及 CK20免疫组化检测。【结果】 本实验NOD-Scid小鼠组的第3代和第5代皮下移植瘤的病例成瘤率分别为92%、58%,BALB/C裸小鼠组的分别为75%、50%。病理切片显示肿瘤组织细胞排列成不规则腺体状,有病理性核分裂相。CDX-2、CK20染色呈阳性。【结论】初步建立了临床背景清楚的人结肠癌的NOD-Scid小鼠和BALB/C裸小鼠皮下移植瘤模型,为进一步研究结肠癌发病机理及治疗策略奠定基础。     

紫外线诱导SKH-1无毛小鼠皮肤鳞癌模型的构建及DNA-PKcs在小鼠及成人皮肤组织的表达

目的 紫外线照射SKH-1小鼠构建皮肤鳞状细胞癌动物模型, 探讨紫外线诱发皮肤SCC发生及AK向SCC转化过程中DNA-PKcs-m TORC2/Akt信号转导通路的DNA-PKcs活化水平。方法 将43只SKH-l无毛小鼠随机分成实验组 (33只) 和对照组 (10只) 。实验组采用SUV-1000日光模拟器模拟日光照射SKH-1无毛鼠, 对照组不做任何处理。在第12、18、24、28周分别处死小鼠行皮肤组织病理检查。28周取同一只小鼠背部不同皮损行组织病理检查和免疫组化检测。收集成人日光性角化病 (AK) 、皮肤鳞状细胞癌 (SCC) 、正常避光部位 (NNS) 、正常曝光部位 (NES) 皮肤组织各5例, 进行后续Western Blot验证。结果 12周后实验组部分小鼠背部皮肤增厚, 皮棘隆起;17周开始实验组小鼠背部皮肤陆续出现直径≥l mm的丘疹;照射28周后成瘤率达100%;对照组未见肿瘤形成。DNA-PKcs和p-DNA-PKcs (T2609) 均细胞定位于细胞质和细胞核, 小鼠表皮组织中DNA-PKcs表达含量增加, 统计学分析有差异 (P<0.05) , 而p-DNA-PKcs (T2609) 无差异 (P>0.05) 。人表皮组织中DNA-PKcs和p-DNA-PKcs (T2609) 表达含量均增加且有统计学分析差异 (P<0.05) 。结论 成功制备小鼠AK模型和SCC模型;明确了紫外线诱导皮肤鳞状细胞癌的过程中, 可诱发DNA损伤及修复等过程。     

维甲类化合物R8605的抗化学致癌和抗促癌作用

本实验根据癌发生的多阶段理论,利用化学性诱发的小鼠二阶段皮肤乳头瘤模型。研究了维甲类化合物R8605的抗化学致癌和抗促癌作用。结果表明,R8605(50mg/kg和25mg/kg,口服)能明显抑制由二甲基苯蒽(DMBA)和巴豆油诱发的小鼠皮肤乳头瘤生成,并表现出较强的抗促癌作用;另外,R8605还能明显抑制巴豆油对小鼠皮肤表皮鸟氨酸脱羧酶(ODC)活性的诱导。R8605较大剂量(100mg/kg,口服)时,也能抑制巴豆油诱发的小鼠耳部肿胀。R8605的抗促癌作用可能与其抑制ODC活性及抗炎作用有一定的关系。     

小鼠早期肺腺癌模型的建立

目的：构建小鼠早期肺腺癌模型,为影像学研究提供工具。方法：210只昆明小鼠随机5个实验组（每组各40只）和1个对照组（10只）。实验组每周皮下注射亚硝基胍（MNNG）溶液0.2 ml（浓度2.0 mg/ml）,按总剂量分为A、B、C、D、和E组,分别注射1、2、4、8和12周。于首次注射后的第60、80、100、120天在每组中随机抽取10只处死,解剖肺脏并观察肿瘤鼠发生率、肿瘤数量和大小等。对照组每周皮下注射0.2 ml的生理盐水共12周。结果：1、成瘤率和成瘤总数与时间、剂量呈正相关,但首次注射第60天时,实验组均无肿瘤生成。2、第80天时,A、B、C、D、E组的成瘤率分别为0%、10%、30%、40%和50%,成瘤总数分别为0、4、 42、60和81个,直径＜0.5 mm的肿瘤数分别为0、4、25、31和40个。3、第100天时,每组的成瘤率分别为20%、40%、100%、100%和100%,成瘤总数分别为6、19、187、223和301个,直径＜0.5 mm的肿瘤数分别为5、16、132、124和123个。4、第120天时,每组的成瘤率分别为30%、50%、100%、100%、100%,成瘤总数分别为30、124、302、351和362个,直径＜0.5 mm的肿瘤数分别为21、98、123、140和108个。经病理证实这些肺肿瘤均为肺腺癌。结论：浓度为2.0 mg/ml的亚硝基胍溶液0.2 ml每周皮下注射连续四周,100天时能获得100%的肺腺癌成瘤率和最多的＜0.5 mm肿瘤数,为最佳的小鼠早期肺腺癌模型。     

复方卡力孜然酊联合NB-UVB治疗儿童白癜风临床疗效观察

目的:观察外涂复方卡力孜然酊联合NB-UVB照射治疗儿童白癜风的疗效和安全性。方法:治疗组26例,于患处外涂复方卡力孜然酊,每天2次,行NB-UVB局部照射,隔日1次。对照组26例,于患处外涂复方卡力孜然酊,每天2次。结果:治疗组有效率100%,对照组有效率92.31%。结论:复方卡力孜然酊联合NB-UVB局部照射治疗儿童白癜风安全、有效,值得临床推广。     

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.