Quantitative evaluation of cardiopulmonary functional reserve in treated patients with pulmonary embolism

Background  There is no research, either at home or abroad, focusing on assessing the cardiopulmonary functional reserve and exercise tolerance in patients with pulmonary embolism (PE), but the benefits of early exercise are well recognized. The goals of this study were to assess cardiopulmonary functional reserve in treated PE patients using the inert gas rebreathing method of the cardiopulmonary exercise test (CPET), and to compare it with traditional methods.

Methods  CPET on the bicycle ergometer were performed in 40 patients with age, gender, body mass index, systolic blood pressure, and pulmonary function matched. The first group was the PE group composed of 16 PE patients (5 male, 11 female) who were given the standard antithrombotic therapy for two weeks. The second group was composed of 24 normal individuals (10 male, 14 female). Both groups were evaluated by cardiac ultrasound examination, 6-minute walking test (6MWT), and CPET.

Results  (1) Right ventricular systolic pressure (RVSP) in the PE group increased significantly compared to the control group, (34.81±8.15) mmHg to (19.75±3.47) mmHg (P <0.01). But neither right atrial end-systolic diameter (RASD) nor right ventricular end-diastolic diameter (RVDD) in the PE patients had changed when compared with the controls. The 6-minute walk distance was significantly reduced in the PE patients compared with normal subjects, (447.81±79.20) m vs. (513.75±31.45) m (P <0.01). Both anaerobic threshold oxygen consumption (VO₂AT) and peak oxygen consumption (VO₂peak) were significantly lower in patients with PE, while CO₂ equivalent ventilation (VE/VCO₂ slope) was higher; VO₂AT (9.44±3.82) ml∙kg^-1∙min^-1 vs. (14.62±2.93) ml∙kg^-1∙min^-1 (P <0.01) and VO₂peak (12.26±4.06) ml∙kg^-1∙min^-1 vs. (23.46±6.15) ml∙kg^-1∙min^-1 (P <0.01) and VE/VCO₂ slope 35.47±6.66 vs. 26.94±3.16 (P <0.01). There was no significant difference in resting cardiac output (CO) between the PE and normal groups, whereas peak cardiac output (peak CO) and the difference between exercise and resting cardiac output (ΔCO) were both significantly reduced in the PE group; peak CO (5.97±2.25) L/min to (8.50±3.13) L/min (P <0.01), ΔCO (1.29±1.59) L/min to (3.97±2.02) L/min (P <0.01). (2) The 6-minute walk distance did not correlated with CPET except for the VO₂ peak in patients with PE, r=0.675 (P <0.01).

Conclusions  The cardiopulmonary functional reserve was reduced in patients with PE. CPET is an accurate, quantitative evaluation of cardiopulmonary functional reserve for PE patients.

     

CD4+T淋巴细胞在小鼠病毒性心肌炎发生发展中的作用

目的探讨CD4⁺ T淋巴细胞（CD4⁺ T细胞）在小鼠柯萨奇病毒B₃(coxsackievirus, CVB₃)所致病毒性心肌炎发生发展中的作用。方法随机将70只BALB/c健康雄性小鼠分为正常对照组(n=15)和病毒感染组(n=55)，正常对照组小鼠腹腔接种不含病毒的eagle氏液0.15mL，病毒感染组小鼠腹腔接种0.15mL TCID50 为109/mL的CVB₃ Nancy株病毒液。分别于接种病毒后第7、10、14、21、28d随机取正常对照组小鼠3只、病毒感染组小鼠8只断髓处死，无菌条件下取心脏，光镜下观察心肌组织的病理变化并计算心肌病理组织学积分，应用免疫组化方法检测CD4⁺ T细胞在心肌中的表达，并与心肌病理组织学积分作相关性分析。结果病毒感染组小鼠心肌可见CD4⁺ T细胞的表达，且与心肌病理组织学积分成显著正相关（r=0.98，P＜0.01），不同时期CD4⁺ T细胞表达强弱不同，以第10～14d最强（P＜0.01)。结论CD4⁺ T细胞在小鼠病毒性心肌炎发病过程中，清除病毒的同时还引起了自身心肌损伤。     

1,25-Dihydroxyvitamin D3 pretreatment enhances the efficacy of allergen immunotherapy in a mouse allergic asthma model

Background  Allergen-specific immunotherapy can induce immune tolerance to specific allergens by regulating immune status of individuals. However, its clinical application is limited due to individual differences in efficacy among patients and un-confirmed safety. 1,25 Dihydroxyvitamin D₃ (1,25(OH)₂D₃) has been shown to be involved in a variety of physiological processes, including immune response regulation. In the present study we explored the role of 1,25(OH)₂D₃ pretreatment for immunotherapy.

Methods  Seventy-five BALB/c mice were randomly divided into five groups (15 mice per group). The mouse allergic asthma model was established by intra-peritoneal injection of ovalbumin (OVA, 10 μg) and aluminium hydroxide (2 mg) as an adjuvant. Intra-peritoneal injection of 50 ng of 1,25(OH)₂D₃ served as a pretreatment, subcutaneous injection of OVA (100 μg) as an immunotherapy, and 1% OVA inhalation as a challenge. Histopathological analysis was performed on four mice per group. The number of cells and their classification in bronchoalvolar lavage (BAL) fluid were assayed. Levels of serum OVA-specific immunoglobulin E (sIgE) and IFN-γ, IL-4, IL-5 and IL-10 in BAL fluid were measured by ELISA.

Results  After 1,25(OH)₂D₃ pretreatment, immunotherapy could significantly inhibit the infiltration of inflammatory cells into lung tissues and BAL fluid of mice with allergic asthma when compared with un-treated animals (eosinophils: (7.46±1.34)×10⁴/ml vs. (13.41±1.67)×10⁴/ml, P <0.05). In addition, levels of IL-4 ((36.91±7.87) pg/ml vs. (43.70±6.42) pg/ml, P >0.05) and IL-5 ((41.97±7.93) pg/ml vs. (60.14±8.35) pg/ml, P <0.05) in BAL fluid and serum sIgE ((0.42±0.05) vs. (0.75±0.06) OD units, P <0.05) were profoundly reduced. However, the IL-10 level in BAL fluid was significantly increased ((67.74±6.57) pg/ml vs. (44.62±8.81) pg/ml, P <0.05).

Conclusions  These results indicated that 1,25(OH)₂D₃ pretreatment enhanced the inhibitory effects of immunotherapy on allergic airway inflammation. In the treatment of allergic diseases, 1,25(OH)₂D₃ pretreatment may be beneficial for improving the efficacy of immunotherapy. 

     

HPLC-MS/MS测定人血浆阿奇霉素浓度及生物等效性研究

目的 建立人血浆中阿奇霉素的高效液相色谱/质谱/质谱(HPLC-MS/MS)测定法,测定受试者口服阿奇霉素颗粒后的血药浓度,并对受试制剂与参比制剂的生物等效性进行评价。方法 19名健康受试者采用随机双交叉试验设计,单剂量口服阿奇霉素颗粒受试制剂和参比药物各500 mg,用HPLC-MS/MS测定用药后不同时间血药浓度。血药浓度-时间数据经DAS 2.0统计软件处理,计算主要药动学参数,并进行两种制剂的生物等效性评价。结果 受试制剂和参比制剂的T_max分别为(2.5±1.1)h和(2.6±1.7)h,C_max分别为(574.6±209.2) ng·mL^-1和(594.5±229.9) ng·mL^-1,t_1/2分别为(44.7±15.2) h和(42.0±13.0) h,AUC_0-144分别为(5 319.6±2 507.8) h·ng·mL^-1和(5 710.7±2 710.1)h·ng·mL^-1,AUC^_0-∞分别为(5 704.2±2 858.7) h·ng·mL^-1和(6 010.0±2 808.1) h·ng·mL^-1, 以AUC_0-144计算,相对生物利用度为(94.2±15.7)%。两制剂的主要药动学参数无显著性差异。结论 受试制剂与参比药物生物等效。     

CD4+ CD25+ Foxp3+及CD4+ CD25- Foxp3+调节性T细胞在小鼠衰老过程中的变化规律

目的分析小鼠脾细胞中CD4⁺ CD25⁺ Foxp3⁺及CD4⁺ CD25^- Foxp3⁺调节性T细胞随小鼠年龄增长的变化, 探讨调节性T细胞与衰老的关系。方法2个月(青年)、6个月(中年)和15个月(老年)小鼠各5只，无菌取脾脏制备单个脾细胞悬液，首先用PECy5-CD4抗体和FITC-CD25抗体进行细胞表面染色，PBS洗涤后，经打孔液和固定液处理，最后用PE-Foxp3抗体进行核内染色，并用流式细胞仪检测CD4⁺调节性T细胞的变化。结果脾脏中CD4⁺ T细胞的数量在不同年龄小鼠间变化不大，CD4⁺ T细胞中CD4⁺ CD25⁺ T细胞的数量也无明显改变，但随小鼠年龄的增长CD4⁺ Foxp3⁺ T细胞的数量明显升高，15个月小鼠CD4⁺ Foxp3⁺ T数量高达（16.83±0.44）%，明显高于2个月和6个月小鼠（P＜0.01）。随着年龄的增长CD25⁺ Foxp3⁺调节性T细胞数量逐渐减低，CD25⁺ Foxp3⁺ T细胞在15个月和6个月小鼠脾细胞中的数量明显低于2个月小鼠（P＜0.05）；而CD25^- Foxp3⁺ 调节性T瞎胞的数量随着年龄的增长持续升高(P＜0.01)；随着年龄的增长小鼠脾细胞中CD4⁺ CD25⁺ Foxp3⁺ T细胞与CD4⁺ CD25^- Foxp3⁺ T细胞的比值呈下降趋势。结论小鼠衰老过程中CD4⁺ CD25⁺ Foxp3⁺ T细胞的数量明显下降，而CD4 ⁺CD25^- Foxp3⁺ T细胞的数量明显上升，提示这两群调节性T细胞亚群在小鼠衰老中可能发挥不同作用。     

羊心脏浦肯野纤维膜钠泵活动的测量

采用细胞内Na⁺选择电极方法,先以不同频率电刺激驱动羊心脏浦肯野纤维标本,使细胞内Na⁺活度(aⁱN+a)上升到不同高度,在停搏即刻测量Na⁺逐出速率(d(a_Naⁱ)/dt)。进而将所测得d(a_Naⁱ)/dt值相对于各时刻a_Naⁱ作相关曲线(SP-IS:Sodium Pump-Internal Sodium),比较其斜率可以了解膜钠泵活动的相对强弱。这种方法可以较迅速、简便地在生理状态下反复测算膜钠泵活动速率。进而用外差法找到相关曲线与纵坐标交点(Na_ia⁺),它反应了静息态时Na⁺内流速率。还观察了pH值变化、高或低K⁺,高或低Ca²⁺及Mn²⁺、异丙基肾上腺素等对膜钠泵活动速率的影响。     

复方苯磺酸氨氯地平/盐酸贝那普利片人体药动学研究

目的 研究健康受试者单剂量与多剂量口服复方苯磺酸氨氯地平/盐酸贝那普利片后的药动学。 方法 LC-MS/MS测定单剂量与多剂量给药后氨氯地平与贝那普利及贝那普利拉血药浓度并利用DAS软件计算药动学参数。结果 单剂量给药氨氯地平与贝那普利及贝那普利拉的主要药动学参数分别是：t_1/2为(47.3±10.6),(1.3±0.4)和(4.5±0.6) h,C_max为(6.4±1.5),(136.5±40.2)和(158.3±46.7)μg·L^-1,AUC_0-t为(267.7±88.4),(144.3±46.7)和(891.4±265.4)μg·h·L^-1;多剂量给药氨氯地平与贝那普利的主要药动学参数分别是：t_1/2为(45.1±8.7),(1.4±0.4)和(5.3±0.8)h,C_max为(8.2±1.8),(142.4±47.5)和(165.2±40.8)μg·L^-1,AUC_0-t为(413.5±102.4),(155.7±52.8)和(915.7±316.9)μg·h·L^-1,R为(1.6±0.6)、(1.0±0.1)和(1.2±0.1)。结论 复方苯磺酸氨氯地平/盐酸贝那普利片2组分及活性代谢产物在健康受试者体内的吸收速率和消除速度不随连续给药变化,但连续给药后苯磺酸氨氯地平在体内有轻微蓄积。     

曲马氨酚片的人体药物动力学研究

目的 测定曲马氨酚片中曲马多和对乙酰氨基酚的药动学参数。方法 采用高效液相色谱法分别测定20名健康志愿者口服曲马氨酚片（2个剂量组：1片和2片,每组10名）后血清中的药物浓度,进行药动学分析。结果 2组健康志愿者口服曲马氨酚片后的主要药动学参数：曲马多AUC_{0-24 h}（ng·h·mL^-1）分别为2 724.89 ± 1 016.54,1 361.61 ± 441.79;AUC_0-∞（ng·h·mL^-1）分别为3 065.49 ± 1 190.66,1 555.04 ± 582.51;t_max（h）分别为1.8 ± 0.75,1.9 ± 0.57;t_1/2（h）分别为7.34 ± 1.39,7.63 ± 2.02;Kel（h^-1） 分别为0.098 ± 0.019,0.097 ± 0.027; Clr（mL·min^-1）分别为31.84 ± 13.65,30.03 ± 9.20;MRT（h）分别为7.62 ± 1.07,7.77 ± 0.75。对乙酰氨基酚的AUC_{0-24 h}（μg·h·mL^-1）分别为 40.28 ± 10.36,18.37 ± 3.84;AUC_0-∞（μg·h·mL^-1）分别为41.63 ± 10.96,18.81 ± 4.06;t_max（h）分别为0.9 ± 0.46,0.9 ± 0.39;t_1/2（h）分别为5.39 ± 1.16,4.96 ± 1.03;Kel（h^-1）分别为0.13 ± 0.03,0.15 ± 0.03;Clr（mL·min^-1）分别为17.17 ± 4.57,18.42 ± 3.89;MRT（h）分别为4.86 ± 0.48,4.50 ± 0.53。结论 各剂量组之间的主要药动学参数（t_max,t_1/2,Kel,Clr,Vd,MRT)无显著性差异,各剂量组AUC_0-t/dose,AUC_0-∞/dose,C_max/dose比较无显著性差异;符合线性动力学特征。     

Correlation of eosinophil counts in induced sputum and fractional concentration of exhaled nitric oxide and lung functions in patients with mild to moderate asthma

Background  The airway inflammation could be assessed by some noninvasive approaches. To investigate the value of eosinophil counts in induced sputum and fractional concentration of exhaled nitric oxide (FENO) for the regimen adjustment in patients with asthma, the correlation was analyzed between the two parameters and lung function parameter (forced expiratory volume in one second (FEV₁)).

Methods  Sixty-five outpatients with mild to moderate non-exacerbation asthma from Beijing Chao-Yang Hospital were enrolled as treatment group. Combined medications of inhaled corticosteroids plus long-acting beta-2 agonist were administered for one year. Lung function parameters, eosinophil counts in induced sputum, concentration of exhaled nitric oxide and the Asthma Control Test scores were recorded, at regular intervals in the follow-up period. Twenty-one healthy volunteers were enrolled as control group and underwent examination of eosinophil counts in induced sputum, lung function and concentration of exhaled nitric oxide.

Results  Sixty-three subjects from treatment group completed follow-up period for one year or longer. Mean FEV₁ value of the 63 subjects was (2.75±0.54) L at baseline, (2.97±0.56) L and (3.07±0.52) L at month 3 and month 6, respectively, and maintained as (3.14±0.51) L in the following six months. Mean FENO decreased from (61±25) parts per billion (ppb) at baseline to (32±19) ppb at month 3 (P <0.05), and continued to decrease to (22±12) ppb at month 6, the difference being significant when compared to both baseline and control group ((13±8) ppb). Mean eosinophil counts decreased to (0.032±0.011) ×10⁶/ml at month 3, which was significantly different from baseline ((0.093±0.023) ×10⁶/ml) and the control group ((0.005±0.003) ×10⁶/ml (both P <0.05). The eosinophil counts in induced sputum correlated positively with concentration of FENO in the first six months (all P <0.05). The concentration of FENO had a significant negative correlation with FEV₁ value (all P <0.05) in any time point in the follow-up period. The Asthma Control Test scores were 18±5, 19±7, 23±2, 24±1 and 24±1 at months 1, 3, 6, 9 and 12, respectively, which were significantly different from the score at baseline (14±3) (P <0.05 ). The most rapid clinical effect was observed at the second month after treatment.

Conclusion  Eosinophil counts in induced sputum and FENO are sensitive parameters to detect airway inflammation and may be useful in evaluating the efficacy of treatment and adjusting medication regimens.

     

硝苯吡啶对无并发症的高血压病患者疗效观察

10例无并发症的高血压病患者于口服国产硝苯吡啶10mg 3次/d后第3d及1周检测血压及应用超声心动图检测左心功能。血压由21.5±3.4/12.4±1.4kPa(161.4±25.9/92.8±10.8mmHg)分别降至18.7±1.6/11.3±1.2kPa(139.8±11.8/84.6±9.4mmHg)、17.9±1.7/11.3±1.2kPa(134.2±12.7/84.6±8.7mmHg)(P均<0.01);平均动脉压由15.2±0.9kPa(114.2±7.7mmHg)分别降至13.9±0.9kPa(104.2±7.3mmHg)、13.6±0.8kPa(102.0±6.3mmHg)(P均<0.01),总外周阻力由1741.0±392.5dyn·s/cm⁵分别降至1514.0±175.5(P>0.05)及1263.1±237.6dyn·s/cm⁵(P<0.01)。舒张早期流速积分由7.2±1.3分别增至8.4±2.6及8.7±2.1((P均<0.05),舒张早期及心房收缩期峰值流速、心指数等指标亦有所改善。提示国产硝苯吡啶能有效持久降压并改善左心功能。     

80例正常人体表标测Q-T_d~p的观察

采用９１ 导联体表电位标测法（ Ｂ Ｓ Ｐ Ｍ） 观察８０ 例正常人 Ｑ Ｔｐｄ（ Ｑ 波起点到 Ｔ 波峰值间期的离散度） ,与同步１２ 导联心电图 Ｑ Ｔ 离散度（ Ｑ Ｔｄ） 对照,探讨 Ｂ Ｓ Ｐ Ｍ 条件下测定 Ｑ Ｔｄ 的可行性。结果：① Ｑ Ｔｐ 、 Ｑ Ｔｐｄ 、 Ｑ Ｔｐｃ１ｄ（ Ｂａｚｚｅｔｔ 公式校正） 、 Ｑ Ｔｐｃ２ｄ（ Ｆｒｉｄｅｒｃｉａ 公式校正） 分别为（２９２ ．６９ ±３７ ．７５） ｍｓ 、（３６ ．７７ ±７ ．４０） ｍｓ 、（４０ ．２３ ±９ ．０４） ｍｓ 和（４０ ．１１ ±７ ．７３） ｍｓ ;② Ｑ Ｔｐｄ 与 Ｑ Ｔｄ 、 Ｑ Ｔｐｃ１ｄ 与 Ｑ Ｔｃ１ｄ 、 Ｑ Ｔｐｃ２ｄ 与 Ｑ Ｔｃ２ｄ 有良好的相关性;③３ 个年龄组（１８ ～３５ 岁、３６ ～６０ 岁、６１ ～７０ 岁） 各参数间无显著性差异;④ Ｑ Ｔｐ 女性明显大于男性（ Ｐ＜ ０ ．０５） ,其余各值男女性之间均无显著性差异。提示： Ｂ Ｓ Ｐ Ｍ 测定 Ｑ Ｔｐｄ 简便可行、较为准确,正常值可暂定为５２ ｍｓ 。     

肌苷衍生物的碱基核糖化研究

目的研究N1-核糖肌苷衍生物的合成反应。方法用丙叉保护肌苷2′,3′-OH,单甲氧基三苯甲基保护5′-OH,在碳酸钾、18-冠醚-6作用下与2,3,5-O-三苄基-1-溴代核糖反应。结果反应得到N1-核糖肌苷衍生物。结论用酰基做保护基易发生酰基重排反应,改用丙叉保护肌苷2′,3′-OH,再用单甲氧基三苯甲基保护5′-OH与2,3,5-O-三苄基-1-溴代核糖在碳酸钾18-冠醚-6作用下得到目的物N1-(2,3,5-O-三苄基-D-核糖)-2′,3′-O-异亚丙基-5′-O-单甲氧基三苯甲基肌苷。     

阿尔茨海默病大鼠模型PET在体成像研究

目的研究18F-FDG及Aβ蛋白显像剂(4’-schiff-O[11CH3])PET成像技术在判别阿尔茨海默病大鼠模型中的作用。方法对10只注射Aβ1～40复制阿尔茨海默模型大鼠及10只注射生理盐水对照大鼠行HE染色及刚果红染色,检测大鼠脑内病理学改变。用18F-FDG及4’-schiff-O[11CH3]PET显像技术在体观察2组大鼠脑内Aβ蛋白分布及葡萄糖代谢情况。结果模型组大鼠海马出现神经元减少并形成淀粉样斑块。模型组PET显像可见注射侧海马葡萄糖代谢减低及4’-schiff-O[11CH3]摄取增加。对照组仅见轻微神经元损伤,PET显示葡萄糖代谢轻度减低。结论18F-FDG及4’-schiff-O[11CH3]PET显像结合行为学及病理学观察可作为检测模型成功与否的方法之一。     

小剂量多巴酚丁胺门控心肌显像预测左心室功能受损的冠心病患者CABG术后早期疗效

目的用小剂量多巴酚丁胺结合99mTc-甲氧基异丁基异腈(MIBI)门控心肌显像(SPECT)检测结果预测存活心肌在接受冠状动脉旁路移植术(CABG)的左心室功能受损的冠心病患者的疗效。方法对38例接受CABG的患者手术前行静息与小剂量多巴酚丁胺结合99mTc-MIBISPECT显像,术后3个月行静息SPECT随访。根据术后左心室射血分数(LVEF)与基线的变化,患者被分为2组:A组19人,术后LVEF值提高≥5%;B组19人,术后LVEF值提高<5%。结果A组术后LVEF值较基线提高(P<0.001),左心室舒张末期容积(EDV)、左心室收缩末期容积(ESV)明显缩小(P<0.05);B组术后LVEF、EDV、ESV值均无明显改善(P>0.05)。临床随访过程中B组有3位患者因心力衰竭再次住院治疗。以术后LVEF较基线提高≥5%作为标准,用ROC曲线得出多巴酚丁胺负荷试验过程中LVEF提高≥4.5%为预测值。多巴酚丁胺负荷试验过程中LVEF提高≥4.5%在A组中有10人,在B组中有3人。以多巴酚丁胺负荷试验过程中LVEF提高≥4.5%作为术后LVEF提高的标准,敏感度为76.9%,特异性为64%,准确性为68.4%。多巴酚丁胺负荷试验过程中LVEF与术后LVEF有明显相关性(r=0.83,P=0.000);多巴酚丁胺负荷试验过程中EDV、ESV与术后EDV、ESV有明显相关性(r=0.79,P=0.000,r=0.88,P=0.000)。结论小剂量多巴酚丁胺结合99mTc-MIBISPECT显像中LVEF提高≥4.5%可作为术后LVEF提高的预测指标。     

慢性肾小球肾炎患者血清Ⅳ型胶原水平的变化及意义

为探讨慢性肾小球肾炎患者血清Ⅳ型胶原 (CⅣ )质量浓度的变化及其与肾功能损害的关系 ,采用酶联免疫法对 34例慢性肾小球肾炎患者和 30例正常人血清CⅣ水平进行检测。结果 :慢性肾炎患者血清CⅣ质量浓度为( 4 0 1 .4 6± 1 1 8.5 3) μg/L ,明显高于对照组的 ( 6 4 .32± 4 2 .5 4 ) μg/L(P <0 .0 1 )。CⅣ与血 β2 MG、尿 β2 MG、血肌酐(SCr)均呈显著正相关 ,相关系数r分别为 0 .6 97、0 .6 4 4、0 .5 2 3,P均 <0 .0 1 ;与肾小球滤过率 (GFR)呈显著负相关(r=-0 .70 1 ,P <0 .0 1 )。提示 :监测慢性肾小球肾炎患者的血清CⅣ有助于监测、判断疾病的活动及病程进展     

Alu-LTR PCR方法检测HAART治疗系列各细胞亚群的整合型HIV-1 DNA

目的应用Alu-LTR PCR方法对高效抗反转录病毒治疗(highly active antiretroviral therapy,HAART)系列的CD4+T,CD8+T及B细胞进行检测,明确不同细胞亚群及HAART治疗的不同阶段是否存在整合的HIV-1 DNA。方法收集20例HIV-1感染患者HAART治疗过程中0、4、12周及10例健康对照的抗凝血标本,纯化CD4+T,CD8+T及B细胞并提取DNA,应用Alu-LTR PCR方法进行检测。结果 20例HIV-1感染者HAART治疗系列中CD4+T细胞及CD8+T细胞成功扩出目的片段,CD8+T细胞所扩条带亮度均低于CD4+T细胞,HAART治疗系列0、4、12周标本所扩条带亮度没有明显差异。B细胞及10例健康者均为阴性。结论 CD4+T及CD8+T细胞存在整合型HIV-1 DNA,HIV储藏库主要存在于CD4+T细胞内。B细胞内不存在整合型HIV-1 DNA。随着HAART治疗的进程,整合型HIV-1 DNA仍然存在,进一步证明HAART不能根除HIV储藏库。     

APP17肽对Aβ导致神经元毒性作用的影响

通过观察β 淀粉样肽 (Aβ)前体蛋白 (APP1 7)中 3 1 9 3 3 5肽段 (APP1 7肽 )对Aβ引起神经毒性作用的影响 ,进一步证实APP1 7肽的神经营养作用。用固相法合成APP1 7肽、Aβ2 5 3 5,高效液相纯化 ,以人神经母细胞瘤株SY5Y为细胞模型 ,分为正常对照组、Aβ2 5 3 51 0 μmol/L损伤组和Aβ2 5 3 51 0 μmol/L加APP1 7肽 1 0 μmol/L保护组。以细胞计数、噻唑蓝 (MTT)代谢率、LDH漏出率、细胞轴突长度、胞体面积、NT 3免疫细胞化学染色和细胞内游离钙离子(Ca2 + )浓度为观察指标。结果 :与正常对照组相比 ,Aβ2 5 3 5损伤组轴突长度和胞体面积均缩小 ,细胞计数减少 ,MTT代谢率降低 ,LDH漏出率升高 ,细胞内Ca2 + 浓度升高 ,而加入APP1 7肽保护后 ,可使上述指标恢复或接近正常。提示 :APP1 7肽具有神经营养和神经保护作用 ,可减轻Aβ引起的神经元损伤。     

分光光度法测定仙鹤草总酚含量的方法研究

目的建立测定仙鹤草中总酚含量的测定方法。方法通过可见分光光度法,以没食子酸为标准品,采用Folin-Ciocalteus试剂,在装有仙鹤草样品的10mL容量瓶中,依次加入Folin-Ciocalteus试剂0.5mL,20%Na2CO31.7mL,在室温条件下放置90min,然后在波长760nm和660nm测定吸光度。结果总多酚浓度在0～10.4 mg/L与吸光度有良好的线性关系。760nm时的回归方程为y=0.091.8x+0.012.9,R2=0.999 3,加标回收率为98.4%～99.8%;660nm时的回归方程为y=0.084 7x-0.002 6,R2=0.999 7,加标回收率为101.2%～101.9%。结论在760nm和660nm 2个波长处测定的结果,都具有良好的精密度、重复性和回收率,可以作为测定仙鹤草多酚定量分析的参考方法。     

食蟹猴上肢取食执行时间的随龄变化及其与~(99m)Tc-TRODAT-1摄取率的相关关系研究

目的研究食蟹猴随年龄增长伴随的用上肢取食执行时间的变化及其与纹状体99mTc-TRODAT-1〔锝99-2β-[N,N'-双(2-巯乙基)乙撑二胺基]甲基,3β-(4-氯苯基)托烷〕摄取率之间的相关关系。方法选取4、10和15岁3个年龄组的健康食蟹猴共30只,利用改良的运动活动平板定量测定上肢取食执行时间,各年龄组分别选取4只动物用多巴胺转运体(dopamine transporter,DAT)配体99mTc-TRODAT-1结合的单光子发射体层摄影术(single-photon emission computed tomography,SPECT)显像观察脑内纹状体多巴胺转运体放射性摄取率(99mTc-TRODAT-1摄取率)的变化。结果随着年龄的增长,食蟹猴用上肢取食执行时间逐渐延长,纹状体99mTc-TRODAT-1放射性摄取率呈逐渐减低的趋势,直线回归分析结果显示食蟹猴上肢取食执行时间与年龄呈统计学相关关系,且与纹状体99mTc-TRODAT-1放射性摄取率亦呈统计学的相关关系。进一步的独立于年龄的部分相关分析表明年龄增长驱动2者的相关。结论正常食蟹猴随年龄增长脑内99mTc-TRODAT-1摄取率的减少是造成食蟹猴上肢取食执行时间延长的关键因素之一。     

TPRC3通道对MPP~+所致MN9D细胞损伤的保护作用

目的探讨经典瞬间受体电位通道蛋白(transient receptor potential-canonical channel,TRPC)家族在1-甲基-4-苯基吡啶离子(1-Methyl-4-phenylpy ridinium ion,MPP+)导致的MN9D细胞毒性损伤中的作用。方法用不同浓度(62.5、125、250、500、1000μmol/L)的MPP+处理MN9D细胞24h,500μmol/L MPP+处理MN9D细胞不同时间(3、6、12、24、36h),建立细胞损伤模型。用RT-PCR的方法检测MN9D细胞中内源性TRPC的表达情况;500μmol/L MPP+作用于MN9D细胞,以Western blot法检测MN9D细胞内源性TRPC表达的变化;构建GFP与TRPC3-GFP腺病毒载体,转染到MN9D细胞中,以四甲基偶氮唑盐比色法(methods the tetrazolium,MTT)的方法检测过表达的TRPC3对MPP+引起的MN9D细胞损伤的保护作用。结果500μmol/LMPP+处理MN9D细胞6h就可以显著降低细胞内TRPC3蛋白水平的表达,但对TRPC6的表达无影响。过表达TRPC3能够保护MN9D细胞免受MPP+的毒性损伤。结论MPP+特异性地降低MN9D细胞TRPC3蛋白水平,过表达TRPC3可以抑制MPP+引起的MN9D细胞损伤。这种现象提示TRPC3通道可能参与了帕金森病发病的分子机制。