胰岛素样生长因子-Ⅰ对兔关节软骨细胞增殖及代谢的影响

目的　探讨胰岛素样生长因子 - (IGF- )对体外生长的关节软骨细胞分裂增殖及功能代谢的影响。方法　采用体外单层培养的方法 ,应用兔关节软骨细胞 ,对照组培养液为 10 %小牛血清 DMEM,实验组在培养液DMEM中加入 IGF- 使其终浓度分别为 3、10、30、10 0及 30 0 ng/ ml,作用细胞 2、4和 6天 ,检测细胞 DNA含量和基质中糖醛酸的含量。结果　 IGF- 在 3～ 30 0 ng/ ml浓度范围能明显增加培养软骨细胞的 DNA及糖醛酸的含量 ,且以30～ 10 0 ng/ ml作用 4天刺激效果最明显 ,与对照组相比 ,有统计学意义 (P<0 .0 1)。结论　 IGF- 对体外培养软骨细胞有刺激 ,并以剂量时间依赖性方式影响增殖及功能代谢。     

IGFl对兔关节软骨细胞增殖的影响及其形态学观察

目的 :了解胰岛素样生长因子1 (IGF1 )对体外生长的兔关节软骨细胞增殖以及细胞超微结构的影响。方法 :应用兔关节软骨细胞体外单层培养 ,对照组培养液为含 1 0 %小牛血清DMEM ,实验组在培养液DMEM中加入IGFl使其终浓度为 1 0 0 μg·L- 1 ,作用细胞 1周 ,得出细胞生长曲线 ,并分别测得两组DNA含量。同时观察IGFl 作用后 ,软骨细胞超微结构的变化。结果 :结果显示IGFl能明显促进培养软骨细胞增殖。实验组DNA含量 ,与对照组相比 ,有统计学意义 (P<0 .0 1 )。在电镜下 ,细胞主要表现为增殖性改变 ,说明细胞代谢活跃。结论 :IGFl 可刺激软骨细胞增殖 ,细胞超微结构表现代谢活跃状态     

EGF和IGF对体外培养兔关节软骨细胞的影响

目的　了解表皮生长因子 (EGF)和胰岛素样生长因子 (IGF)对体外培养兔关节软骨细胞存活数目和分裂增殖百分数的影响。方法　体外培养兔关节软骨细胞传至第 2代 ,分别以EGF、IGF ,以及二者不同的浓度组合作用于软骨细胞。通过酶联免疫检测仪 (MTT)测定软骨细胞存活数 ,流式细胞仪测定软骨细胞分裂增殖百分数。结果　不同浓度EGF对兔关节软骨细胞存活和分裂增殖均有促进作用 ,作用浓度强度次序为 :10ng/ml>0 1ng/ml>10 0ng/ml。不同浓度IGF对兔关节软骨细胞的存活和分裂增殖均有较强促进作用 ,浓度为 5 0ng/ml时 ,刺激效果最显著。EGF与IGF联合使用时 ,刺激效果优于任何一种因子单独使用 ,而以EGF 10ng/ml和IGF 5 0ng/ml为最佳浓度组合。 结论　EGF和IGF都可促进兔关节软骨细胞存活和分裂增殖 ,但以协同作用效果最佳。     

碱性成纤维细胞生长因子转染兔关节软骨细胞的研究

目的　探讨碱性成纤维细胞生长因子 (bFGF)基因转染兔关节软骨细胞后对培养的关节软骨细胞形态、分裂增殖及代谢等方面的影响。方法　将bFGF基因克隆于真核表达载体pHβ。AP 1中 ,构建重组真核表达载体 pHβ bFGF ,转染兔关节软骨细胞。G418筛选阳性克隆 ,检测阳性细胞bFGF基因的表达水平。测定培养软骨细胞的DNA含量、糖醛酸含量、软骨细胞增殖情况及进行细胞周期分析。结果　bFGF基因转染软骨细胞表型未见显著变化 ;bFGF基因转染组、载体对照组、空白对照组DNA含量分别为 ( 77.37± 6 .2 1)、( 40 .39± 4.33)、( 33 .77± 4.2 5 ) μg/瓶 (P <0 .0 1) ,糖醛酸含量分别为 ( 30 8.8± 10 .2 )、( 77.9± 8.7)、( 80 .2± 10 .5 ) μg/瓶 ( P <0 .0 1) ,软骨细胞G1期分别为 5 9.3± 2 .1、6 9.5± 4.0、73 .1± 3 .9(P <0 .0 5 )。结论　bFGF转染关节软骨细胞后 ,可显著促进细胞分裂增殖并缩短细胞周期 ,为软骨组织工程研究提供新的技术路线及理论基础。     

不同强度持续静态压力对兔腰椎小关节软骨细胞活性影响的实验研究

目的 探讨不同强度持续静态压力对体外培养的兔腰椎小关节软骨细胞活性的影响. 方法 分离新西兰白兔腰椎小关节软骨细胞,体外培养.将培养的第3代软骨细胞分为4组:对照组、30 kPa组、60 kPa组和90 kPa持续静态加压组.显微镜下观察其形态,分别用免疫组化、噻唑蓝(MTT)、酶联免疫吸附法(ELISA)法鉴定软骨细胞,检测细胞增殖及Ⅱ型胶原的合成情况. 结果 第3代软骨细胞Ⅱ型胶原免疫组织化学染色,显示为阳性.软骨细胞增殖OD值,1d除了对照组与30 kPa组相比差异无统计学意义外,其他各组两两比较差异均有统计学意义(P ＜0.05);2～4 d:除了 30 kPa组与60 kPa组相比差异无统计学意义外,其他各组两两比较差异均有统计学意义(P＜0.05);5～10 d:各组间两两比较差异均有统计学意义(P＜0.05).空白对照组的Ⅱ型胶原含量最高[(7.517±0.328)μg/L],其次是60 kPa组[(6.035±0.075) μg/L],90 kPa组含量最低[(2.873±0.127) μg/L],30 kPa组的Ⅱ型胶原含量为(4.846±0.093)μg/L,各组间两两比较差异均有统计学意义(P＜0.05).结论 兔腰椎小关节软骨细胞在持续静态压力下容易发生退变,细胞增殖率和Ⅱ型胶原含量均下降.     

IGF1对兔关节软骨细胞增殖的影响及其形态学观察

目的了解胰岛素样生长因子1(IGF1)对体外生长的兔关节软骨细胞增殖以及细胞超微结构的影响.方法应用兔关节软骨细胞体外单层培养,对照组培养液为含10%小牛血清DMEM,实验组在培养液DMEM中加入IGFl使其终浓度为100μg*L-1,作用细胞1周, 得出细胞生长曲线,并分别测得两组DNA含量.同时观察IGFl作用后,软骨细胞超微结构的变化.结果结果显示IGFl能明显促进培养软骨细胞增殖.实验组DNA含量,与对照组相比,有统计学意义 (P<0.01).在电镜下,细胞主要表现为增殖性改变,说明细胞代谢活跃.结论IGFl可刺激软骨细胞增殖,细胞超微结构表现代谢活跃状态.     

生长因子促成年兔关节软骨细胞增殖的研究

目的　研究促进成年兔关节软骨细胞迅速增殖的方法。方法　将体外培养 2 4～ 2 8月龄的新西兰大白兔的原代关节软骨细胞中加入不同浓度的胰岛素样生长因子 - (IGF- )、碱性成纤维细胞生长因子 (b FGF)及 IGF- b FGF联合应用。于 2 4、4 8及 72小时分别进行 MTT检测软骨细胞的成活数 ;检测传 3代后各组软骨细胞的总增殖倍数 ;第 14天后 ,用流式细胞仪检测各组软骨细胞的增殖倍数 ,观察 IGF- 、b FGF及 IGF- b FGF联合应用对成年兔关节软骨细胞各时间点的促增殖作用。结果　 1施加不同浓度的 IGF- 、b FGF或两者联合应用后各时间点软骨细胞活细胞数明显高于对照组 (P<0 .0 1) ;2传 3代后 ,三组总增殖倍数也都明显高于对照组 (P<0 .0 1) ,且两种因子联合应用 ,增殖倍数明 显比应用单个因子时高 ,有统计学意义 (P<0 .0 1) ;3培养 14天后 ,增殖指数明显高于对照组 (P<0 .0 1) ,两种因子联合应用时 ,增殖指数明显比应用单个因子时高 (P<0 .0 5 ) ;而施加两次因子时 ,增殖指数比施加一次因子时高 (P<0 .0 5 )。结论　 IGF- 、b FGF对成年兔关节软骨细胞有明显的促增殖作用 ,两者之间有协同效应。     

IGF-Ⅰ对培养兔关节软骨细胞作用的实验研究

目的 了解胰岛素样生长因子I（ICF－I）对培养关节软骨细胞增残及代谢的影响。方法 体外单层培养兔关节软骨细胞,实验组以10,100,200ng/mlICF-I作用细胞2,4,6天,对照组不加IGF－I作用培养细胞。检测细胞DNA及基质中糖醛酸含量,用流式细胞仪分析在100ng/mlIGF-I作用下细胞周期亚时相。结果 IGF－I在10－100mg/ml浓度范围内,对培养软骨细胞增残和代谢有促进作用,以100ng/ml浓度作用效果最佳,细胞周期分析,实验组DNA合成前期（G1期）较对照组缩短。结论 IGF－I促进软骨细胞的增殖与代谢,且通过缩短细胞G1期而缩短的细胞周期。     

胰岛素样生长因子对体外培养豚鼠肋软骨细胞的影响

目的：了解胰岛素样生长因子(insulin-1ike growthfactor-1，IGF-1)对体外培养豚鼠肋软骨细胞分裂增殖及功能代谢的影响。方法：体外单层培养豚鼠肋软骨细胞，对照组培养液为无血流清DMEM，实验组在培养液DMEM中加入IGF-1使其终浓度分别为10ng／ml、50ng／ml、100ng／ml，作用6天后，检测细胞DNA含量和培养液中糖醛酸的含量。结果：IGF-1在10～100ng／ml浓度范围能明显增加培养软骨细胞的DNA及糖醛酸的含量，且以50ng／ml作用效果最明显，与对照组相比，有统计擘意义(P＜0．01)。结论：IGF-1对体外培养肋软骨细胞有刺激，并以剂量依赖性方式影响细胞的增殖及功能代谢。     

肝细胞生长因子对大鼠颌下腺细胞增殖的影响

目的　研究肝细胞生长因子 (HGF)对体外培养大鼠颌下腺细胞增殖的剂量 效应及时间 效应。方法　采用体外颌下腺细胞培养在含 1 0 %胎牛血清的合成培养液 (DMEM) ,实验组分别加入不同浓度的HGF ,利用噻唑蓝 (MTT)比色法检测细胞的增殖能力 ,研究HGF对颌下腺细胞增殖的影响。结果　在 1 0 %胎牛血清配制的DMEM含HGF(1 .0～ 1 0 0 0 .0 μg/L)可促进培养细胞的增殖 (P <0 .0 1 ) ,且在一定浓度范围内HGF(1 .0、5 .0、1 0 .0、50 .0、1 0 0 .0 μg/L)呈剂量 效应关系 ,最大效应剂量为 1 0 0 .0 μg/L。加入HGF第 3天开始显示出明显的促增殖效应。 结论　HGF在 1 0 %胎牛血清DMEM培养条件下能够促进颌下腺细胞的增殖和分化 ,经体外条件培养的颌下腺细胞可以作为组织工程学研究中的细胞来源     

Vasopressor hormone response following mesenteric traction during major abdominal surgery

Background : We investigated the vasopressor hormone response following mesenteric traction (MT) with hypotension due to prostacyclin (PGI₂) release in patients undergoing abdominal surgery with a combined general and epidural anesthesia. Methods : In a prospective, randomized, placebo-controlled study we administered 400 mg ibuprofen (i.v.) in 42 patients scheduled for abdominal surgery. General anesthesia was combined with epidural anesthesia (T4-L1). Before as well as 5, 15, 30, 45, and 90 min after MT we recorded plasma osmolality, hemodynamics and measured 6-keto-PGFlα (stabile metabolite of PGI₂), TXB₂ (stabile metabolite of thromboxane A₂) active renin, and arginine vasopressin (AVP) plasma concentrations by radioimmunoassay. Catecholamine levels were assessed by high-pressure liquid chromatography (HPLC) with electrochemical detection. Results : Following MT, arterial hypotension occurred along with a substantial PGI₂ release. This was completely abolished by ibuprofen administration. Although plasma levels of 6-keto-PGF_1α (1133 (708) vs. 60 (3) ng/L, median (median absolute deviation), P=0.0001, placebo vs. ibuprofen) remained significantly elevated, blood pressure was restored within 30 min after MT in the placebo group. At the same point in time plasma concentrations of TXB² (164 (87) vs. 58 (1) ng/L, P=0.0001), epinephrine (46 (33) vs. 14 (6) ng/L, P=0.001), AVP (41 ± (18) vs. 12 (7) ng/L, P=0.0004), and active renin (27 (12) vs. 12 (4) ng/L, P = 0.001) were significantly higher in placebo-treated patients. Conclusion : Under combined general and epidural anesthesia arterial hypotension following MT due to endogenous PGI₂ release is associated with enhanced release of AVP, active renin, epinephrine and thromboxane A₂, presumably contributing to hemodynamic stability within 30 min after MT.     

A Teaspoon Pump for Pumping Blood with High Hydraulic Efficiency and Low Hemolysis Potential

Abstract: Virtually all blood pumps contain some kind of rubbing, sliding, closely moving machinery surfaces that are exposed to the blood being pumped. These valves, internal bearings, magnetic bearing position sensors, and shaft seals cause most of the problems with blood pumps. The original teaspoon pump design prevented the rubbing, sliding machinery surfaces from contacting the blood. However, the hydraulic efficiency was low because the blood was able to "slip around" the rotating impeller so that the blood itself never rotated fast enough to develop adequate pressure. An improved teaspoon blood pump has been designed and tested and has shown acceptable hydraulic performance and low hemolysis potential. The new pump uses a nonrotating "swinging" hose as the pump impeller. The fluid enters the pump through the center of the swinging hose; therefore, there can be no fluid slip between the revolving blood and the revolving impeller. The new pump uses an impeller that is comparable to a flexible garden hose. If the free end of the hose were swung around in a circle like half of a jump rope, the fluid inside the hose would rotate and develop pressure even though the hose impeller itself did not "rotate"; therefore, no rotating shaft seal or internal bearings are required.     

Microspheres Based Detoxification System: A New Method in Convective Blood Purification

Abstract: A variety of protein-bound or hydrophobic substances, accumulating as a result of pathologic conditions such as exogenous or endogenous intoxications, are removed poorly by conventional detoxification methods because of low accessibility (hemodialysis), insufficient adsorption capabilities (hemosorption), low efficiency (peritoneal dialysis), or economic limitations (high-volume plasmapheresis). Combining advantages of existing methods with microspheric technology, a module-based system was designed. Major operating parameters of the latter can be modified to allow for adjustment to individual clinical situations. An extracorporeal blood circuit including a plasmafilter is combined with a secondary high-velocity plasma circuit driven by a centrifugal pump. Different microspheric adsorbers can be combined in one circuit or applied in sequence. Thus, a prolonged treatment can be tailored using specially designed selective adsorber materials. Comparing this system with existing methods (high-flux hemodialysis, molecular adsorbent recycling system), results from our in vitro studies and animal experiments demonstrate the superior efficiency of substance removal.     

Tissue perfusion in relation to cardiac output during continuous positive-pressure ventilation and administration of propranolol or verapamil

Background : Our objective was to determine whether administration of propranolol or verapamil modifies the hemodynamic adaptation to continuous positive-pressure ventilation (CPPV), in particular the regional distribution of cardiac output (CO).
Methods : General hemodynamics and regional blood flows assessed by microsphere technique (15 (μm) were recorded in 16 anesthetized pigs during spontaneous breathing (SB) and CPPV with 8 cm H₂O end-expiratory pressure (CPPV8) before and after intravenous administration of propranolol (0.3 mg · kg⁻¹ followed by 0.15 mg · kg⁻¹ · h⁻¹, n=8) or verapamil (0.1 mg · kg⁻¹ followed by 0.3 mg · kg⁻¹ · h⁻¹, n=8).
Results : CPPV8 depressed CO by 25% without shifts in its relative distribution with the exception of a noteworthy increase in adrenal perfusion. Propranolol increased arterial blood pressure, and due to a fall in heart rate, CO dropped by 25%. The kidneys and, to a lesser extent, the splanchic region and central nervous system received increased fractions of the remaining CO at the expense of skeletal muscle flow. Similar patterns were seen during SB and CPPV8 such that the combination of propranolol and CPPV8 depressed CO by 50%. The circulatory effects of verapamil were less evident but myocardial perfusion tended to increase.
Conclusions : The combination of propranolol or verapamil with CPPV does not result in any specific hemodynamic interaction in anesthetized pigs, except that the combined effect of propranolol and CPPV may severely reduce CO.     

Bariatric Surgery in Some "Central and East-European (former Eastern bloc)"Countries - Current Status and Prediction for the Next Millennium

Background: Obesity is increasing globallly, including in the formerly "Eastern Bloc" countries. Methods: A survey was made of obesity and bariatric surgery. Results: In the 8 East and Central European countries studied, with total population 300 million, roughly 43% of the population was overweight (BMI 25-30), 23% obese (BMI > 30), with about 15 million people morbidly obese (BMI > 40). From 0-10 morbidly obese individuals/100,000/year undergo bariatric surgery. Conclusion: Most countries were found to provide inadequate treatment for obesity.The majority of the morbidly obese are not treated effectively. However, health-care awareness of obesity and bariatric surgeons are slowly increasing.     

Volatile anaesthetics reduce adhesion of blood platelets under low-flow conditions in the coronary system of isolated guinea pig hearts

Background : Inhibitory effects of volatile anaesthetics on platelet aggregation have been demonstrated in several studies. However, the influence of volatile anaesthetics on intracoronary platelet adhesion has not been elucidated so far.
Methods : Isolated hearts of guinea pigs were perfused with buffer in the absence or presence of volatile anaesthetics (0.5 and 1 MAC) at constant coronary flow rates of 5 ml/min for 25 min, then 1 ml/min for 30 min and again 5 ml/min for 10 min. Before, during and after low-flow perfusion, a bolus of human platelets was applied into the coronary system. To simulate thrombogenic conditions, 0.3 U/ml human thrombin was infused during low-flow perfusion and reperfusion. The number of platelets sequestered to the endothelium was calculated from the difference between coronary in- and output of platelets. The myocardial production of lactate and consumption of pyruvate and coronary perfusion pressure were also determined.
Results : At a flow rate of 5 ml/min only about 3% of the applied platelets did not emerge from the coronary system, in any group. In contrast, 13.1±1.2% (mean±SEM) of infused platelets became adherent in low-flow perfusion in the control group without anaesthetic. The adherence was reduced with each 1 MAC isoflurane (to 6.2±1.2%), sevoflurane (to 4.4±0.9%) or halothane (to 3.2±1.5%) (each P <0.05 vs. control). Volatile anaesthetic, 0.5 MAC, did not inhibit platelet adhesion to a statistically significant extent in any case. Perfusion pressure and metabolic parameters were not statistically different between the control and the hearts exposed to anaesthetics.
Conclusion : Volatile anaesthetics in a concentration of 1 MAC can reduce the adhesion of platelets in the coronary system under reduced flow conditions. This action does not arise from vasodilation or inhibition of ischaemic stress.     

Synergistic interaction between the effects of propofol and midazolam with fentanyl on phrenic nerve activity in rabbits

Background: It has been shown that the depressive effects of both propofol and midazolam on consciousness are synergistic with opioids, but the nature of their interactions on other physiological systems, e. g. respiration, has not been fully investigated. The present study examined the effect of propofol and midazolam alone and in combination with fentanyl on phrenic nerve activity (PNA) and whether such interactions are additive or synergistic. Methods: PNA was recorded in 27 anaesthetised and artificially ventilated rabbits. In three groups, propofol, fentanyl and midazolam were administered intravenously in incremental doses to construct dose-response curves for the depressant effects of each one on PNA. In another two groups, the effect of pretreatment with either fentanyl 1 μg · kg^?1 i. v. or midazolam 0.05 mg · kg^?1 i. v. on the effects of propofol and fentanyl respectively on PNA were studied. Results: Propofol and fentanyl caused a dose-dependent depression of PNA with complete abolition at the highest total doses of 16 mg · kg^?1 i. v. and 32 μg · kg^?1 i. v., respectively. In contrast, midazolam in incremental doses to a total of 0.8 mg · kg^?1 reduced mean PNA by 63%, but approximately 12% of PNA remained at a total dose as high as 6.4 mg · kg^?1. The mean ED₅₀s, calculated from dose-response curves, were 5.4 mg · kg^?1, 3.9 μg · kg^?1 and 0.4 mg · kg^?1 for propofol, fentanyl and midazolam, respectively. Initial doses of either fentanyl 1 μg · kg^?1 i. v. or midazolam 0.05 mg · kg^?1 i. v. acted synergistically with subsequent doses of either propofol or fentanyl to abolish PNA at total doses of 8 mg · kg^?1 and 8 μg · kg^?1, respectively. Conclusion: Fentanyl has a synergistic interaction with both propofol and midazolam on PNA and hence potentially on respiration.     

Influence of dopexamine hydrochloride on haemodynamics and regulators of circulation in patients undergoing major abdominal surgery

Background: Catecholaminergic support is often used to improve haemodynamics in patients undergoing major abdominal surgery. Dopexamine is a synthetic vasoactive catecholamine with beneficial microcirculatory properties. Methods: The influence of perioperative administration of dopexamine on cardiorespiratory data and important regulators of macro- and microcirculation were studied in 30 patients undergoing Whipple pancreaticduodenectomy. The patients received randomized and blinded either 2 μg · kg^?1 · min^?1 of dopexamine (n=15) or placebo (n=15, control group). The infusion was started after induction of anaesthesia and continued until the morning of the first postoperative day. Endothelin-1 (ET-1), vasopressin, atrial natriuretic peptide (ANP), and catecholamine plasma levels were measured from arterial blood samples. Measurements were carried out after induction of anaesthesia, 2 h after onset of surgery, at the end of surgery, 2 h after surgery, and on the morning of the first postoperative day. Results: Cardiac index (CI) increased significantly in the dopexamine group (from 2.61±0.41 to 4.57±0.78 1 · min^?1 · m^?2) and remained elevated until the morning of the first postoperative day. Oxygen delivery index (DO₂I) and oxygen consumption index (VO₂I) were also significantly increased in the dopexamine group (DO₂I: from 416±91 to 717±110 ml/m² · m²; VO₂I: from 98±25 to 157±22 ml/m² · m²), being significantly higher than in the control group. pHi remained stable only in the dopexamine patients, indicating adequate splanchnic perfusion. Vasopressive regulators of circulation increased significantly only in the untreated control patients (vasopressin: from 4.37±1.1 to 35.9±12.1 pg/ml; ET-1: from 2.88±0.91 to 6.91±1.20 pg/ml). Conclusion: Patients undergoing major abdominal surgery may profit from prophylactic perioperative administration of dopexamine hydrochloride in the form of improved haemodynamics and oxygenation as well as beneficial influence on important regulators of organ blood flow.     

Systemic analgesia adapted to the children's condition

A concept of balanced analgesia using nonsteroidal anti-inflammatory drugs (NSAIDs), paracetamol (acetaminophen), opioids, and corticosteroids can also be used in patients with pre-existing illnesses. NSAIDs are the most effective treatment for acute pain of moderate intensity in children; however, these drugs should be avoided in patients at increased risk for serious side effects, e.g. patients with renal impairment, bleeding tendency, or extreme prematurity. NSAIDs can be given with minimal risks to the younger child with mild to moderate asthma, and, in these patients, the use of steroids can be encouraged; in addition to their antiemetic and analgesic action, a beneficial effect on asthma symptoms can be expected. In the non-intubated child with cerebral trauma, exaggerated sedation caused by opioids and increased bleeding tendency caused by NSAIDs must be avoided. In neonates and small infants, the oral administration of sucrose or glucose is helpful to minimize pain reaction during short uncomfortable interventions.