Retroviral vector containing human p16 gene and its inhibitory effect on Bcap -37 breast cancer cells

目的　研究p16基因对肿瘤细胞生长抑制及细胞周期阻滞作用。方法　将p16cDNA插入逆转录病毒载体pLXSN构建成p16基因逆转录病毒重组体pLp16SN。利用基因转染方法 ,将pLp16SN及pLXSN导入逆转录病毒包装细胞系PA317细胞 ,获得逆转录病毒。用逆转录病毒感染Bcap 37乳腺癌细胞 ,经G4 18筛选获阳性克隆。利用Northern和Westernblotting方法检测p16基因的表达。检测转基因细胞的生长速度 ,细胞周期及裸鼠成瘤等细胞生物学行为的改变。结果　Northern及Westernblotting显示转染p16基因的Bcap 37细胞p16基因mRNA及蛋白质表达明显增高。此细胞较未转染基因的Bcap 37细胞和转染空载体的Bcap 37细胞生长速度慢 ,G1期细胞比率增高 ,裸鼠接种成瘤体积小。结论　p16基因高表达能够抑制乳腺癌细胞Bcap 37的生长 ,并阻滞细胞从G1期进入S期     

沙利霉素对口腔鳞癌细胞增殖和凋亡的影响

目的:探讨沙利霉素(salinomycin)对口腔鳞癌细胞增殖和凋亡的影响,并初步探讨沙利霉素对信号通路的影响。方法:培养口腔鳞状细胞癌细胞系CAL-27,将1、2、4、8、16、32 μmol/L沙利霉素和1.25、2.5、5、10、20、40、80 μmol/L顺铂与CAL-27细胞共同培养,24 h和48 h后用细胞计数试剂盒-8(cell counting kit-8,CCK-8)法检测沙利霉素和顺铂对CAL-27细胞增殖的影响;0、2、4、8 μmol/L沙利霉素和0、5、10、20 μmol/L顺铂与CAL-27细胞共培养48 h后,通过流式细胞术检测沙利霉素和顺铂对CAL-27细胞周期的影响,蛋白免疫印迹法(Western blot)检测CAL-27细胞中天冬氨酸特异性半胱氨酸蛋白酶-3(cysteine-containing aspartate-specific proteases-3,Caspase-3)、天冬氨酸特异性半胱氨酸蛋白酶-9(cysteine-containing aspartate-specific proteases-9,Caspase-9)、脱氧核糖核酸(deoxyribonucleic acid,DNA)修复酶(poly ADP-ribose polymerase,PARP)、蛋白激酶B(protein kinase B, Akt)和磷酸化蛋白激酶B (phosphorylated protein kinase B,p-Akt)的表达。结果:CCK-8实验表明沙利霉素和顺铂均能显著抑制口腔鳞状细胞癌CAL-27细胞增殖,且抑制作用呈时间依赖性和药物浓度依赖性,但是相对于临床一线化疗药物顺铂而言,沙利霉素对CAL-27细胞增殖的抑制效果更加显著(P<0.001)。细胞周期检测表明,与加入二甲基亚砜(dimethyl sulfoxide,DMSO)的对照组相比,8 μmol/L沙利霉素与CAL-27细胞共同培养48 h后,细胞休眠期/DNA合成前期的CAL-27细胞比例明显升高(40.40%±1.99% vs.64.46%±0.90%,P<0.05), DNA合成期和DNA合成后期/有丝分裂期的CAL-27细胞比例出现降低(24.32%±2.30% vs.18.73%±0.61%,P<0.05,35.01%±1.24% vs.16.54%±1.31%,P<0.05);顺铂对CAL-27细胞周期没有特异性改变。蛋白免疫印迹法结果显示,沙利霉素在上调CAL-27细胞中Caspase-3和 Caspase-9蛋白表达(P<0.05)的同时下调PARP、Akt和p-Akt蛋白的表达(P<0.05)。结论:相对于顺铂而言,沙利霉素对CAL-27细胞增殖有更强的抑制作用,并且能将口腔鳞状细胞癌CAL-27细胞周期阻滞在细胞休眠期/DNA合成前期,同时能够诱导CAL-27细胞发生凋亡,这一机制可能和Akt/p-Akt 信号通路相关。     

足背皮神经的显微断层解剖及其临床意义

目的 为不同部位断足再植提供理论基础和有关数据。方法 用体视学方法对６６例成人足背皮神经进行了观测。结果 腓深神经的平均横径和面积分别为：第１断层３．０５ｍｍ,３．２８ｍｍ＾２;腓浅神经的平均横径和面积分别为：第１断层３．１４ｍｍ,２．３８ｍｍ＾２;足背内侧皮神经的平均横径和面积分别为：第１断层２．７５ｍｍ,２．１４ｍｍ＾２,第５断层２．６８ｍｍ,２．０９ｍｍ＾２;足背中间皮神经的平均横径和面积分     

Hyposensitivity of peripheral α-adrenoceptors in respiratordependent amyotrophic lateral sclerosis assessed by intravenous norepinephrine infusion

Intravenous norepinephrine infusion test was performed in eight patients with amyotrophic lateral sclerosis (ALS) supported by respirators and nine control subjects, to examine -adrenoceptor function of peripheral resistant blood vessels. Baseline plasma norepinephrine concentrations in ALS patients were significantly higher than those in control subjects, indicating basal sympathetic hyperactivity (normal 218.2 ± 59.7 pg/ml; ALS 450.0 ± 288.4 pg/ml). The stimulus-response curves in the patients were similar to those in control subjects, and there were no significant differences between mean gains of the stimulus—response curves in both groups (normal 18.7 ± 5.5; ALS 15.2 ± 11.2). However, three ALS patients, two of whom had circulatory fluctuation and sympathetic hyperactivity, revealed lower gain levels than the mean minus 2 SD in control subjects (4.7, 1.1 and 3.7). This indicates hyposensitivity or down-regulation of the -adrenoceptor function of peripheral blood vessels in these ALS patients. For early detection of sympathetic hyperactivity and prediction of circulatory collapse, it would be useful to measure the plasma norepinephrine concentration and the gain of the norepinephrine infusion curve in respirator-dependent ALS patients.     

尼可地尔超极化心脏停搏心肌保护的实验研究

目的：评估尼可地尔（Ｎｉｃｏｒａｄｉｌ）超极化心脏停搏心肌保护的有效性,并与高钾心肌麻痹液去极化心脏停搏的心肌保护效果进行比较。方法：实验分３组,即对照组、去极化心脏停搏组、超极化心脏停搏组。１０ｍｌ４℃心肌麻痹液诱停离体鼠心,每３０ｍｉｎ重复灌注心肌麻痹液５ｍｌ,在（１５±１）℃下心脏停搏１２０ｍｉｎ,再灌注后离体心工作３０ｍｉｎ。记录心脏机械停搏时间、再灌注后心脏复跳情况、心功能指标恢复（左室发展压、主动脉流量及左室压力微分）、冠状循环灌注平均流量、心肌丙二醛（ＭＤＡ）含量、电镜下心肌超微结构的改变。结果：超极化心脏停搏组对心功能恢复、冠状循环灌注平均流量及心肌组织超微结构的保护效果优于去极化心脏停搏组（Ｐ＜０．０５）,心肌ＭＤＡ含量低于去极化心脏停搏组（Ｐ＜０．０１）。结论：尼可地尔超极化心肌麻痹液（１００μｍｏｌ／Ｌ）心肌保护效果优于高钾去极化心肌麻痹液     

Cerebral vasoconstriction in comatose patients resuscitated from a cardiac arrest?

Objective To determine the role of cerebral vasoconstriction in the delayed hypoperfusion phase in comatose patients after cardiac arrest.Design Prospective study.Setting Medical intensive care unit in a university hospital.Patients 10 comatose patients (Glasgow Coma Score 6) successfully resuscitated from a cardiac arrest occurring outside the hospital.Measurements We measured the pulsatility index (PI) and mean blood flow velocity (MFV) of the middle cerebral artery, the cerebral oxygen extraction ratio and jugular bulb levels of endothelin, nitrate, and cGMP during the first 24 h after cardiac arrest.Results The PI decreased significantly from 1.86±1.02 to 1.05±0.22 (p=0.03). The MFV increased significantly from 29±10 to 62±25 cm/s (p=0.003). Cerebral oxygen extraction ratio decreased also from 0.39±0.13 to 0.24±0.11 (p=0.015). Endothelin levels were high but did not change during the study period. Nitrate levels varied widely and showed a slight but significant decrease from 37.1 mol/l (median; 25th–75th percentiles: 26.8–61.6) to 31.3 mol/l (22.1–39.6) (p=0.04). Cyclic guanosine monophosphate levels increased significantly from 2.95 nmol/l (median; 25th–75th percentiles: 2.48–5.43) to 7.5 nmol/l (6.2–14.0) (p=0.02).Conclusions We found evidence of increased cerebrovascular resistance during the first 24 h after cardiac arrest with persistent high endothelin levels, gradually decreasing nitrate levels, and gradually increasing cGMP levels. This suggests that active cerebral vasoconstriction due to an imbalance between local vasodilators and vasoconstrictors plays a role in the delayed hypoperfusion phase.     

Development of totally implantable electromechanical artificial heart systems: Baylor ventricular assist system

An implantable electromechanical ventricular assist system (VAS) intended for permanent use has been developed. It consists of a conically shaped pumping chamber, a polyolefin (Hexsyn) rubber diaphragm attached to a conically shaped pusher-plate, and a compact roller-screw actuator. Design stroke volume is 63 ml. The device weighs 620 g, and has a total volume of 348 ml. The pump can provide 8 L/min flow against 120 mm Hg afterload with a preload of 10 mm Hg. The inner surfaces are biolized by dry gelatin coating, with inflow and outflow ports accommodating tissue valves. Three subacute in vivo validation studies have been conducted in calves up to two weeks. The entire system functioned satisfactorily in both the fill/empty and the fixed-rate modes. There was no thromboembolic complication without anticoagulation. The pump showed reasonable anatomical fit inside the left thorax. This VAS is compact, efficient, quiet, and easy to control.     

Metabolic aspects of maximal exercise performance after slow release metoprolol and after atenolol

Summary Maximal exercise performance by eight healthy male subjects was tested after one week of medication with slow-release metoprolol 200 mg/d (metoprolol-SR), atenolol 200 mg/d or placebo, in a double blind crossover trial. The maximal working capacity was significantly decreased after atenolol and metoprolol-SR. Plasma glucose and FFA concentrations during the exercise test did not change: either after placebo therapy or after beta-blockade. The anaerobic threshold did not change after beta-blockade, but the changes in lactate due to the exercise were less after beta-blockade. Neither beta-blocker affected the exercise-induced alteration in airway resistance. Both drugs caused a small but significant ventilatory depression at rest and at 75% of maximal exercise. It is concluded that the limiting factor in maximal exercise performance after ₁-adrenergic blockade does not lie in oxygen transport to the working muscles via ventilation and the circulation, but is most probably due to anaerobic metabolism.     

急性颅高压心脏停搏类型的临床分析

目的　探讨急性颅高压患者心脏停搏类型。方法　对 2 5例急性颅高压患者进行 2 4 h连续心电监护 ,选择心肺复苏术前 5～ 30 s心电描记波形为心脏停搏类型。结果　 13例急性颅脑损伤患者有 12例表现为缓慢性心律失常—心室停顿。1例为心室颤动 ,12例急性脑中风患者有 11例表现为缓慢性心律失常—心室停顿。1例为电机械分离。结论　缓慢性心律失常—心室停顿是急性颅高压患者心脏停搏的最主要类型     

Design and test of a vascular access device

Transarterial left ventricular assist devices (LVADs), such as the Hemopump, IABP, and PUCA-pump, are meant to be introduced into the body via the femoral or axillary artery without major surgery. For certain applications, introduction is performed directly into the aorta via an open thorax procedure. A prototype of a vascular access device has been realized that allows direct access into the aorta as an alternative for the common surgical graft anastomosis suturing technique. The device consists of a metal tube acting as a circular knife to cut a hole in the aortic wall, a screw to store the removed part of the aortic wall, and a plastic tube that is introduced through the hole and tightly connected to the aortic wall. The device could be placed without aortic clamping. The device has been tested on a slaughterhouse porcine aorta. A low-pressurized aorta appeared to be the worst case; thus, two animal experiments in the low-pressurized pulmonary artery were performed. No leakage occurred for pressures between 40 and 300 mm Hg.