小鼠对HBV DNA疫苗的免疫应答及细胞因子的免疫佐剂效应

目的 :观察编码小鼠 IL- 2及 IL- 12的真核表达载体 ,对 HBV DNA疫苗诱导 Balb/ c小鼠 (H- 2 d )免疫应答的调节作用及其对稳定表达 HBs Ag的小鼠肥大细胞瘤 P815细胞 (P 815 - HBV- S)成瘤性的影响。　方法 :肌内注射HBV DNA疫苗及细胞因子真核表达载体 ;背部皮下接种 P 815 - HBV - S细胞 ,观察成瘤情况 ;EL ISA法测定血清抗HBs;4h5 1 Cr释放法检测小鼠脾细胞 CTL 活性。　结果 :免疫 8周后 ,以 p CR3.1- S、p CR3.1- S+IL- 2及 p CR3.1-S+IL- 12免疫的小鼠血清 ,45 0 nm A值分别为 0 .87± 0 .1、1.98± 0 .17及 1.6 7± 0 .15 ,均较 p CR3.1组显著提高(P<0 .0 5 )。CTL 细胞杀伤活性分别为 (5 0 .5± 6 .4) %、(6 1.9± 7.1) %及 (73.3± 8.8) % ,后两组与 p CR3.1- S组比较差异均显著 (P<0 .0 5 )。脾细胞悬液经抗 CD4+ 单克隆抗体处理后 ,CTL细胞杀伤活性分别为 (48.3± 5 .9) %、(5 6 .2± 7.5 ) %和 (75 .6± 9.1) % ,抗 CD8+ 单克隆抗体处理后分别为 (10 .6± 1.4) %、(13.6± 1.9) %和 (16 .9±2 .3) %。对照组成瘤率为 10 0 % ,p CR3.1- S组小鼠成瘤率为 12 .5 % ,p CR3.1- S+IL- 12真核表达载体组成瘤率为0 ,对照组平均存活期和 6周后存活率分别为 2 8.4天和 0天。后两组分别 >38.2天及 45     

IL-12对乙型肝炎DNA疫苗诱导小鼠免疫应答的影响

目的　观察编码鼠 IL - 12的真核表达载体对 HBVDNA疫苗诱导 BAL B/ c小鼠免疫应答的影响 .方法　肌肉注射 DNA疫苗 ,EL ISA法检测小鼠血清抗 HBs,4h5 1 Cr释放法检测小鼠脾细胞 CTL杀伤活性 .结果　免疫 8wk后 ,单纯注射 p CR3.1- S及共注射 IL - 12真核表达载体的小鼠血清 45 0 nm A值分别为 0 .87± 0 .1及 1.6 7± 0 .15 .CTL 细胞杀伤活性分别为 (5 0 .5± 6 .4) %及 (73.3± 8.8) % ,两组均有明显差异 (P<0 .0 1) ,脾细胞悬液经抗 CD4+ 单克隆抗体处理后 CTL 细胞杀伤活性分别为 (48.3± 5 .9) % ,(75 . 6±9.1) % ,抗 CD8+单克隆抗体处理后分别为 (10 .6± 1.4) % ,(16 .9± 2 .3) % .结论　 IL - 12的真核表达载体能够提高小鼠对 DNA疫苗的免疫应答 ,CTL细胞杀伤活性主要由 CD8+执行 .基因疫苗可能用于预防及治疗 HBV感染     

IL-12的真核表达载体对乙肝基因疫苗的免疫佐剂效应

观察编码鼠IL - 12的真核表达载体对HBVDNA疫苗诱导Balb/c小鼠免疫应答的影响。方法 :肌肉注射DNA疫苗 ,ELISA法检测小鼠血清抗HBs,4h51Cr释放法检测小鼠脾细胞CTL杀伤活性。结果 :免疫8周后 ,注射 pCR3.1组、注射 pCR3.1-S组及共注射IL - 12真核表达载体的血清OD值分别为 0 .0 4± 0 .0 1、0 .87± 0 .1及 1.6 7± 0 .15。CTL细胞杀伤活性分别为 (10 .1± 6 .4) %、(5 0 .5± 6 .4) %及 (73.3± 8.8) % ,后两组与pCR3.1组比较均有显著差异 (P <0 .0 0 1) ,后两组比较亦有显著差异 (P <0 .0 5 )。脾细胞悬液经抗CD4 单克隆抗体处理后分别为 (48.3± 5 .9) %、(75 .6± 9.1) % ,抗CD8 单克隆抗体处理后分别为 (10 .6±1.4) %、(16 .9± 2 .3) %。结论 :IL - 12的真核表达载体能够提高小鼠对DNA疫苗的免疫应答 ,CTL细胞杀伤活性主要由CD8 执行。     

IL-12及HBV基因疫苗共同免疫小鼠的效果

目的　观察编码鼠 IL- 12的真核表达载体对 HBV DNA疫苗诱导 BAL B/ c小鼠免疫应答的影响 .方法　肌肉注射 DNA疫苗 ,EL ISA法检测小鼠血清抗 HBs,4h5 1 Cr释放法检测小鼠脾细胞 CTL杀伤活性 .结果　免疫 8wk后 ,注射 p CR3.1组、注射 p CR3.1- S组及共注射 IL- 12真核表达载体的血清 A(OD值 )分别为 (0 .0 4± 0 .0 1) ,(0 .87± 0 .1)及 (1.6 7± 0 .15 ) .CTL 细胞杀伤活性分别为 (10 .1± 6 .4) % ,(5 0 .5± 6 .4) %及 (73.3± 8.8) % ,后两组与 p CR3.1组比较均有显著差异 (P<0 .0 1) ,后两组比较均有显著差异 (P<0…     

乙型肝炎病毒核心区DNA疫苗接种后H-2d小鼠的特异性免疫应答

目的　观察乙型肝炎病毒 (HBV)核心区DNA疫苗接种后BALB/c(H 2 d)小鼠的特异性免疫应答。方法　构建HBV核心区DNA疫苗 (PJW430 3/HBc) ;用基因枪法和肌内注射法将该DNA疫苗接种BALB/c小鼠 ;ELISA法检测小鼠血清抗 HBc(IgG)及IgG亚类 (IgG1,IgG2a) ;51铬释放法检测小鼠脾细胞HBcAg特异性CTL活性。结果　该DNA疫苗在体外转染细胞中可良好表达HBcAg。血清抗 HBc终点滴度在DNA疫苗基因枪组和肌肉注射接种组小鼠分别为 1∶32 80 5 0和 1∶10 935 0。两组小鼠的抗 HBcIgG亚型均以IgG2a略占优势。两组小鼠的HBbAg特异性CTL杀伤活性分别达到 5 1 1%和 5 5 2 %。结论　HBV核心区DNA疫苗在小鼠实验中具有良好的细胞免疫和体液免疫原性。     

MUC1/Y基因疫苗治疗膀胱肿瘤的实验研究

目的探讨人黏蛋白MUC1/Y基因DNA疫苗诱导小鼠特异性细胞毒性T细胞(cytotoxic Tlym-phocytes,CTL)及体液免疫应答的作用,为人膀胱癌疫苗的应用提供基础实验研究资料。方法通过pEGFP-C3-MUC1/Y质粒免疫雄性BALB/c小鼠,采用ELISA法检测小鼠抗体生成情况和CTL分泌IL-2和IFN-γ的量,并用LDH释放法测定CTL对膀胱癌细胞株BIU-87的特异性杀伤率。结果在接种疫苗的小鼠体内特异性抗体明显升高,CTL分泌的IL-2和IFN-γ较对照组高。在效靶比为100∶1、50∶1、25∶1、12.5∶1时,实验组特异性CTL对BIU-87杀伤率分别为50.9%、45.9%、47.5%和18.6%,而对照组(空载体pEGFP-C3和灭菌生理盐水)分别为18.3%、10.7%、13.8%、6.3%;16.5%、11.9%、8.6%和10.3%。实验组与对照组两者之间差异具有显著性(P<0.01)。结论MUC1/YDNA疫苗能够诱导小鼠体内产生抗人膀胱癌黏蛋白的特异性抗体,并能诱导产生特异杀伤表达MUC1/Y的靶细胞的CTL,为MUC1/Y基因疫苗用于膀胱肿瘤生物免疫提供一种新的治疗策略。     

双剂量乙肝免疫球蛋白联合乙肝疫苗阻断乙肝母婴传播的效果调查

目的回顾性调查分析乙肝表面抗原(HBsAg)阳性孕妇所生的婴儿经联合接种乙肝疫苗和双剂量200IU乙肝免疫球蛋白(HBIG)后产生的预防效果。方法研究对象为320例HBsAg阳性孕妇所生的婴儿,出生后联合接种10mg乙肝疫苗(0、1、6个月)和双剂量200IU HBIG(0、2周),出生后第7个月检测婴儿血液HBsAg和抗HBs浓度。结果 2.8%的婴儿(9/320)乙肝免疫预防失败(HBsAg阳性),2.5%的婴儿(8/320)呈免疫应答无反应(抗HBs浓度<10IU/L),94.7%的婴儿(303/320)免疫应答有反应(抗HBs≥10IU/L)。HBeAg阴性和HBeAg阳性孕妇所生的婴儿免疫接种失败率分别为0%和5.17%(P<0.05),HBV DNA阴性和HBV DNA阳性孕妇所生的婴儿免疫接种失败率分别为0%和4.41%(P<0.05),但母亲HBeAg和HBV DNA阳性与婴儿对乙肝疫苗无免疫应答不相关(P>0.05)。结论通过联合乙肝疫苗和双剂量200IUHBIG接种,绝大多数婴儿(94.7%)在出生后第7个月可产生保护性抗体,母亲HBeAg和HBV DNA可影响婴儿的乙肝免疫预防失败率,但与婴儿对乙肝疫苗免疫应答无反应不相关。     

HBsAg核酸疫苗诱导H-2b小鼠体液免疫应答的初步研究

目的：观察adw和adr亚型HBV PreS2 S核酸疫苗单一或联合HBcAg核酸疫苗诱导的小鼠体液免疫反应。方法：C57BL／6小鼠23只随机分为4组;pJW4303组（P组）,pJW4303/S2 S(adw亚型）组（W组）、pJW43/3S2 S(adr亚型）组（R组）,pJW4303/S2 S(adr) pJW4303/HBc组（R＋C组）。免疫方法为每只小鼠每次肌注相应质粒DNA100μg(100μl)。免疫程序为0、2、4周。于第3次免疫后4周W组、R组、R＋C组小鼠血清全部检出抗－HBs,抗体含量随免疫次数增多而增加,抗－HBs浓度最高达22329IU／L,与P组（对照质粒组）相比差异有显著性（P＜0．01）,但各核酸疫苗组之间差异无显著性（P＞0．5）。抗－HBs浓度最高达22329IU／L,与P组（对照质粒组）相比差异有显著性（P＜0．01）,但各核酸疫苗组之间差异无显著性（P＞0．5）。而抗－HBc在第一次免疫后2周即在R＋C组全部出现,抗－HBs和抗－HBc在P组始终未检出。结论：adw和adr亚型HBV PreS2 S核酸疫苗和HBc核酸疫苗能诱导H－2^b小鼠产生较强的体液免疫反应,联合HBc核酸疫苗有助于抗－HBs的早期产生。     

基因疫苗诱导小鼠抗HBV皮下移植瘤免疫研究

目的　观察 HBV DNA疫苗 (p CR3.1- S)诱导 BAL B/ c小鼠 (H- 2 d)的特异性细胞免疫应答及其对稳定表达 HBs Ag的小鼠肥大细胞瘤 P815细胞 (P815 - HBV- S) (H- 2 d)成瘤性的影响 .方法　肌肉注射 DNA疫苗 ,背部皮下接种 P815 - HBV- S细胞 ,观察成瘤情况 ,4h5 1 Cr释放法检测小鼠脾细胞 CTL 活性 .结果　接种 DNA疫苗后小鼠成瘤率为 12 .5 % ,对照组为 10 0 % .小鼠平均存活期大于 38.2 d,对照组为 2 8.4d,40 d后小鼠存活率为 87.5 % ,对照组为 0 % . CTL细胞杀伤活性明显增加 ,p CR3.1- S组5 1% ,对照组为 2 1% (P<0 .…     

乙型肝炎病毒c基因启动子变异与血清HBeAg及HBV DNA的关系

目的 :研究乙型肝炎病毒 (HBV ) c基因启动子 (BCP)变异与 e抗原及 HBV DNA含量的关系。方法 :采用错配引物聚合酶链反应 (PCR) -限制性片段长度多态性 (RFL P)分析及荧光定量 PCR(FQ- PCR)对 10 5例血清 HBV DNA阳性 HBV感染者进行 HBV BCP基因变异及 HBV DNA定量测定。结果 :BCP变异组 HBe Ag阴性率为 84 .4 % (5 4 / 6 4 ) ,非 BCP变异组为2 6 .8% (11/ 4 1)。 BCP变异组 HBV DNA含量在 HBe Ag+ 病例 (10 8.79± 0 .85vs 10 7.4 8± 0 .39copy/ ml,P <0 .0 1)及 HBe Ag- 病例(10 8.0 5± 1 .1 6 vs10 6 .55± 0 .91 copy/ ml,P<0 .0 1)中均较非 BCP变异组高。结论 :HBV BCP变异在下调前 c/ c基因表达的同时可能促进 HBV复制 ,对 HBs Ag+ / HBe Ag- 患者需要进一步检测 HBV DNA,以免由于基因变异导致将 HBe Ag阴性者误认为病毒的免疫清除或静息而延误抗病毒治疗时机     

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.