蓇密钙片与马鹿角多肽对成骨细胞增殖分化的研究

目的观察蓇密钙片及马鹿角多肽对鼠胚成骨细胞MC3T3-E1的增殖分化作用及细胞内OPG、RANKL mRNA的表达。方法采用MEM(10%FBS)培养液培养细胞,细胞计数法描绘成骨细胞生长曲线;MTT法检测蓇密钙片(0、50、100、500、1000μg/ml)、马鹿角多肽(0、5、50、500μg/ml)和钙尔奇D对照组对鼠胚成骨细胞MC3T3-E1增殖的作用;蓇密钙片(0、50、100、500、1000μg/ml)、马鹿角多肽(0、5、50、500μg/ml)和钙尔奇D对照组干预鼠胚成骨细胞MC3T3-E1 72 h后,比色法检测细胞内碱性磷酸酶(alkaline phosphatase,ALP)的含量,RT-PCR法检测骨保护蛋白(osteoprotegerin,OPG)、核因子КB激活受体配体(receptor activator of NK-КB ligand,RANKL)mRNA的表达。结果蓇密钙片及马鹿角多肽作用于MC3T3-E1成骨细胞72 h后,可促进其增殖,OD值增加(P0.05),增加ALP的含量,促进OPG mRNA表达,同时抑制RANKL mRNA表达。结论蓇密钙片及马鹿角多肽促进MC3T3-E1成骨细胞增殖分化,可能与促进成骨细胞OPG mRNA表达、抑制RANKL mRNA表达有关,从而促进骨形成,抑制骨吸收。     

铁饱和乳铁蛋白对大鼠成骨细胞增殖与分化的影响

目的 探讨铁饱和乳铁蛋白对体外培养的大鼠成骨细胞增殖和分化的影响.方法 用酶消化法分离大鼠颅盖骨成骨细胞进行原代培养;乳铁蛋白螯合Fe~(2+)获得铁饱和乳铁蛋白;将其稀释到不同浓度作用于成骨细胞,四唑盐比色法(MTT)测定细胞增殖;碱性磷酸酶法(ALP)测定细胞碱性磷酸酶活性.结果 随着时间的增加,各浓度组均可促进成骨细胞增殖及增强碱性磷酸酶活性.其中100 μg/mL浓度组在72 h时细胞增殖数目最大、碱性磷酸酶活性最高,较对照组有统计学意义(P＜0.01).结论 铁饱和乳铁蛋白能促进大鼠成骨细胞的增殖和分化.     

三七总甙对成骨细胞增殖、分化及0PG表达影响的研究

目的 观察不同剂量的三七总甙对体外培养大鼠成骨细胞增殖、分化及OPG表达的影响，探索体外作用的机制和最佳剂量。方法 第2代培养的成骨细胞分别加入终浓度为0μg／ml、10μg／ml、50μg／ml、100μg／ml的三七总甙，观察细胞生长、钙结节形成情况，碱性磷酸酶(ALP)染色、分泌量检测，骨钙素(OCN)检测，MTT法观察成骨细胞增殖，免疫组化SP法结合阳性细胞计数法检测成骨细胞OPG表达。结果 成骨细胞在7d可铺满瓶壁，30d可形成钙结节，三七总甙可促进成骨细胞的增殖ALP、OCN的分泌，以50μg／ml时作用明显。三七总甙可促进成骨细胞OPG的表达，在50μg／ml时作用明显。结论 三七总甙可促进大鼠成骨细胞的增殖、分化，促进成骨细胞OPG的表达，以50μg／ml时作用明显。     

橄榄苦苷对成骨细胞OPG/RANKL mRNA的影响

目的 通过测定橄榄苦苷对成骨细胞OPG/RANKL mRNA的表达,探讨其治疗骨质疏松症的机制。方法 运用荧光定量PCR技术（RT-PCR）检测不同浓度橄榄苦苷不同时间对成骨细胞OPG/RANKL mRNA 表达量。结果 与阴性对照组比较,RT-PCR 结果显示不同浓度的橄榄苦苷均能促进OPG mRNA的表达,并且抑制成骨细胞RANKL mRNA 的表达。结论 橄榄苦苷可能通过抑制RANKL分泌来降低破骨细胞活性,同时促进成骨细胞分泌OPG,使之与RANKL结合增多,间接作用于破骨细胞,使其活性降低,而达到治疗骨质疏松的目的。     

乳铁蛋白促进成骨细胞增殖的最佳浓度及促进分化的最佳时间

目的 探索乳铁蛋白促进大鼠成骨细胞增殖的最佳浓度及1000μg/mL乳铁蛋白对促进成骨细胞分化的最佳时间。方法 用混合酶消化法分离大鼠颅盖骨成骨细胞进行原代培养;不同浓度乳铁蛋白干预成骨细胞,浓度分别为0、0.1、1、10、100、200、400、600、800和1000μg/mL,在1、3、5、7d后分别采用CCK-8法测定细胞增殖;用1000μg/mL的乳铁蛋白干预成骨细胞7、14、21d后,采用碱性磷酸酶染色法检测细胞内碱性磷酸酶的活性。结果 1.CCK-8法测定细胞增殖结果显示,低于100μg/mL浓度乳铁蛋白组其OD值随着浓度的增高而增高,而高于100μg/mL浓度乳铁蛋白组其OD值随着乳铁蛋白浓度的增高而递减;2.碱性磷酸酶染色法结果显示,1000μg/mL的乳铁蛋白在干预第21天时碱性磷酸酶活性最高。结论 100μg/mL乳铁蛋白是促进大鼠成骨细胞增殖的最佳浓度;1000μg/mL的乳铁蛋白促进成骨细胞分化作用呈时间依赖性。     

牛源乳铁蛋白对体外培养大鼠成骨细胞RANKL和OPG mRNA表达的影响

目的观察牛源乳铁蛋白对成骨细胞核因子κβ受体活化受体（RANKL）／护骨素（OPG）mRNA表达的影响。方法体外分离培养新生大鼠成骨细胞，分别观察不同浓度乳铁蛋白处理不同时间对成骨细胞RANKL／OPGmRNA表达的影响。采用半定量RT-PCR方法检测RANKL／OPGmRNA。结果乳铁蛋白呈时间和剂量依赖性地刺激大鼠成骨细胞OPGmRNA的表达，而抑制RANKLmRNA的表达，使RANKL／OPG值降低。结论乳铁蛋白通过调节成骨细胞RANKL／OPG的基因表达，从而抑制破骨细胞介导的骨吸收。     

联合甲状旁腺激素和普萘洛尔对人成骨细胞增殖及OPG和RANKL mRNA表达的影响

目的 联合甲状旁腺激素(rhPTH 1-34)和普萘洛尔(PRO)处理体外培养的人松质骨源性成骨细胞(HOB),观察其对人成骨细胞增殖及骨保护素(OPO)和核因子KB受体活化因子配体(RANKL )基因表达的影响,探讨其对骨代谢影响的可能机制。方法 (1)以成人骼骨和股骨颈部松质骨为原料,分离培养原代人成骨细胞并对其鉴定。(2)以人成骨细胞为体外实验模型,固定浓度rhPTH1- 34 (50 ng/ml)和不同浓度PRO(0. 1μM,1μM,10μM)分别及联合刺激体外培养的人成骨细胞72 h后,采用CCK-8法检测细胞增殖能力,用实时荧光定量PCR法检测成骨细胞OPG和RANKL基因的表达。结果 rhPTH1-34和PRO单独给药均可促进成骨细胞增殖(P <0.05),在PRO 10μM时成骨细胞OPG基因的表达量增加(P<0.05),且大于RANKL基因的表达量;相反,联合rhPTH 1-34和PRO给药抑制成骨细胞增殖(P<0.05),并抑制成骨细胞OPG和RANKL基因的表达(P<0.05),且随着PRO浓度的增加,OPG和RANKL基因表达均呈下降趋势。结论 rhPTH1- 34和PRO单独给药均可促进成骨细胞增殖,而两者联合给药后却抑制了成骨细胞的增殖,可能是通过调控OPG和RANKL基因的表达来实现的。     

阿仑膦酸钠、氟化钠联合作用对体外培养大鼠成骨细胞的影响

目的探讨阿仑膦酸钠联合氟化钠对体外培养大鼠成骨细胞的作用。方法采用酶消化法分离SD大鼠颅盖骨中成骨细胞,原代培养。分别给予不同浓度的阿仑膦酸钠和氟化钠分组刺激,观察药物对成骨细胞增殖和碱性磷酸酶活性的影响,并用Western Blot法测定骨保护素(osteoprotegerin,OPG)/核因子-B受体活化因子配体(receptor activator for nuclear factor-B ligand,RANKL)蛋白表达。结果阿仑膦酸钠最佳单药浓度为10~(-6) mmol/L,氟化钠的最佳单药浓度为10~(-8) mmol/L,两种药物最佳浓度联合应用更能促进成骨细胞增殖,碱性磷酸酶(alkaline phosphatase,ALP)活性更强,OPG表达升高、RANKL表达下调,与对照组相比,差异有统计学意义。高浓度氟化钠(10~(-3)mmol/L、10~(-4)mmol/L)对成骨细胞有抑制作用。结论阿仑膦酸钠、氟化钠低浓度时均促进成骨细胞增殖和分化,二者联合有协同作用。高浓度氟化钠对成骨细胞有抑制作用。     

右归饮对激素性股骨头坏死大鼠体外诱导培养成骨细胞作用的研究

目的:在右归饮对体外正常大鼠成骨细胞的增殖和分化作用研究基础上,进一步观察右归饮对激素性股骨头坏死大鼠体外诱导培育成骨细胞的增殖和分化以及基因表达的影响。方法:取体重为100～120g雄性SD大鼠20只,臀肌注射醋酸强的松龙49mg/kg·d,连用6d(造模),7d后处死,无菌条件下取造模大鼠骨髓基质细胞,诱导培养成骨细胞,然后随机分为4组:高、中、低浓度右归饮含药血清组(R1、R2、R3)和对照组(R4),分别添加高、中、低浓度右归饮含药血清及对照血清,继续培养72h后,进行成骨细胞增殖、碱性磷酸酶活性、骨保护素(OPG)和破骨细胞分化因子(RANKL)表达水平检测(实时荧光定量PCR检测),并进行统计学分析。结果:高、中浓度右归饮含药血清组(R1、R2)与对照组(R4)相比,成骨细胞增殖率显著升高(P0.01),碱性磷酸酶活性显著升高(P0.01),成骨细胞OPGmRNA的相对表达量显著升高(P0.01),且与浓度剂量呈一定的正相关;成骨细胞RANKLmRNA显著降低(P0.01)。低浓度右归饮含药血清组(R3)与对照组(R4)相比,成骨细胞增殖率、碱性磷酸酶活性以及OPG、RANKLmRNA相对表达量无统计学差异。结论:高、中浓度的右归饮含药血清对体外培养的SINFH大鼠成骨细胞的增殖和分化具有明显的促进作用,并可提高成骨细胞OPGmRNA的相对表达量而对其RANKLmRNA有抑制作用。右归饮含药血清对体外诱导培养成骨细胞的作用与剂量有一定相关性,在一定的浓度范围内促进成骨细胞的分化、增殖。     

黄瓜籽总皂苷对MC3T3-E1细胞分化及SPARC、OPG/RANKL表达的影响

目的观察黄瓜籽总皂苷提取物(cucumber seed saponins,CSS)对小鼠成骨细胞MC3T3-E1增殖、分化和矿化的影响,以及与骨质疏松相关的SPARC、OPG/RANKL/RANK信号通路的作用。方法通过MTT实验、碱性磷酸酶(alkaline phosphatase,ALP)活性检测、茜素红染色,考察不同浓度CSS对MC3T3-E1细胞增殖、分化及矿化的影响;采用RT-PCR方法检测SPARC、OPG/RANKL mRNA表达水平; Western blot检测SPARC、OPG/RANKL的蛋白表达量。结果与阳性对照组相比,CSS能明显促进MC3T3-E1细胞增殖(P0.05),CSS高、中剂量组能明显提高MC3T3-E1细胞ALP活性及钙化结节数量(P0.05);与空白组相比,CSS不同剂量组均明显上调SPARC、OPG/RANKL mRNA及蛋白表达水平(P0.05)。结论黄瓜籽总皂苷能够促进成骨细胞MC3T3-E1的增殖、分化及矿化能力,并通过上调SPARC、OPG/RANKL的表达水平提高MC3T3-E1细胞的成骨能力。     

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.     

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.     

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.     

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.     

Immunomodulation in Transplantation with Photopheresis

Abstract: Photopheresis is a technique in which peripheral blood mononuclear cells, in the presence of a photoacti-vatable compound, are exposed extracorporeally to ultraviolet A light and reinfused, inducing a host autoregula-tory immune response. Experimental work and ongoing clinical studies are helping to define the role of this novel, safe, and non-toxic immunomodulating technology in the field of transplantation.