α-干扰素对人功能性垂体腺瘤细胞的作用

目的 采用体外细胞培养，免疫组织化学染色和放射免疫测定方法，研究α-干扰素（α—IFN）对体外培养人垂体功能性腺瘤细胞激素分泌的影响。方法 对11例垂体生长激素（GH）及GH／催乳素（PRL）腺瘤和15例垂体PRL腺瘤进行体外细胞培养，观察细胞形态的改变和生长特点。采用免疫组织化学染色技术研究α—IFN和αR—IFN在侵袭性和非侵袭性垂体腺瘤、功能性和非功能性垂体腺瘤之间阳性表达率差异。结果 大多数垂体腺瘤细胞有α-IFN和αR—IFN表达，尤其在侵袭性垂体腺瘤细胞中呈高表达；α—IFN对垂体腺瘤细胞激素分泌有较强的抑制作用；α—IFN对大多数溴隐亭（BC）或奥曲肽（SMS）耐药的腺瘤细胞仍有抑制激素分泌效应．α-IFN能增强BC对垂体PRL腺瘤分泌的抑制作用。结论 α—IFN具有用于临床治疗功能性垂体腺瘤的可能性。     

干细胞移植入鼠胶质瘤模型体内后的生物学观察

目的 体外培养扩增大鼠胚胎神经干细胞(NSCs)及骨髓基质细胞(BMSCs),并移植入大鼠胶质瘤模型体内,观察细胞生物学特性改变及其对胶质瘤的影响.方法 取孕15 d的大鼠海马区神经干细胞及大鼠股骨骨髓基质细胞,体外培养扩增;建立胶质瘤模型,待肿瘤生长至3周和4周时,分别随机抽取5只大鼠行MRI检测;分不同时间段大鼠灌注取材,分别做苏木素一伊红(HE)染色及BrdU免疫组织化学染色,观察细胞存活情况及对胶质瘤的影响.结果 两种细胞移植人胶质瘤模型体内均能够大量存活.BMSCs组、NSCs移植组、对照组模型平均生存时间分别为4.03、4.28、3.88周;3周及4周时肿瘤的平均直径分别为(6.4mm/7.6mm、5.4mm/6.0mm、7.2mm/8.0mm).BrdU染色示两种细胞均对胶质瘤细胞有定向靶向作用.结论 神经干细胞对胶质瘤细胞的生长有抑制作用,能够延长大鼠模型的生存期.骨髓基质细胞对胶质瘤生长无抑制作用.两种细胞均对胶质瘤细胞有定向靶向作用,能够作为生物载体转染目的 基因治疗胶质瘤.     

脑肿瘤

颅内高度恶性非生殖细胞瘤性生殖细胞肿瘤；应用替莫唑胺对照司莫司丁治疗恶性脑胶质瘤的疗效观察；胚胎发育不良性神经上皮肿瘤临床及病理分析；三脑室脑膜瘤；脑胶质瘤特异性6HRE-GFAP-Bax α基因治疗系统的构建；垂体泌乳素腺瘤术后复发相关因素分析；神经节细胞胶质瘤（附28例分析）；影响垂体生长激素腺瘤经蝶手术疗效的因素分析     

微囊包膜垂体的研制及移植研究

将ｈＧＨ腺瘤细胞微囊包膜包裹后移植至去垂体大鼠腹腔内，结果，大鼠血清ＧＨ，水平升高，生长加快。但随时间的延长，移植之微包囊垂体功能逐渐消失，在第４周微囊开始有破碎，第８周时完全碎解吸收。提示我们制备的微囊包膜在活体内具有免疫隔离作用，但其寿命不长，仅４周左右。功能性垂体腺瘤可作为垂体移植的供体来源。     

重组生长激素与大鼠模型肝癌增殖关系的研究

目的　研究重组生长激素 (rGH ,思增 )对大鼠移植型肝癌模型肿瘤细胞增殖的影响 ,以了解rGH的使用对治疗肝癌是否有不利影响。方法　用腹腔移植型雄性种鼠的含肿瘤细胞的腹腔液接种于幼鼠的皮下 ,制成皮下种植实体瘤鼠。接种 7d后 ,将实体瘤切成 2mm× 2mm× 3mm大小瘤组织块 ,再将瘤组织块植于 82只成年雄性大鼠的肝上而成移植型肝癌模型。随机均分成实验组和对照组。实验组术后第 3天从尾静脉分别给予rGH 0 .5U / (kg·d)和生理盐水 ,连续 5d。手术后 8,10 ,12d3个时段处死。大鼠、肝脏、肿瘤组织称重 ,全自动图像分析仪检测肝细胞及肿瘤细胞数目密度、核分裂数、核分裂指数 ;术后第 8天处死的大鼠同时行3H TdR活体掺入计数。结果　除术后第 8天对照组大鼠体重较术前明显减轻外 (P <0 .0 5 ) ,两组其他各时段术前术后差异均无显著性 (P >0 .0 5 ) ;肝重、肝细胞及肿瘤细胞数目密度、肿瘤重、核分裂数、核分裂指数 ,同一时段两组间差异均无显著性 (P >0 .0 5 ) ;术后第 8天 3H TdR活体掺入计数 ,两组差异无显著性 (P >0 .0 5 ) ;肿瘤重量在同一组不同时段差异有极显著性 (P <0 .0 1)。结论　静脉注射rGH与大鼠移植肝癌细胞的增殖无明显关系。     

人工毛乳头异种移植诱导大鼠足垫毛囊形成

目的观察人头皮毛乳头细胞微囊(人工毛乳头)异种移植诱导大鼠足垫毛囊形成的能力。方法以海藻酸钠-多聚赖氨酸-海藻酸钠(alginate-polylysine-alginate,APA)微囊包裹分离培养的毛乳头细胞;对体外培养1、4周的毛乳头细胞微囊及无APA的微囊对照组行组织学观察;取培养4周的毛乳头细胞微囊移植至大鼠足垫皮下,6周后取材行组织学检查。结果毛乳头细胞微囊体外培养1周后,毛乳头细胞周围出现细胞外基质;4周后,囊中形成“类毛乳头样结构”;人头皮毛乳头细胞微囊移植至大鼠足垫6周后,移植部位及其周围皮下有大量毛囊及皮脂腺结构形成。结论人工毛乳头诱导并参与了无毛区域新生毛囊及皮脂腺的组织构成。     

Angiostatin基因治疗人肝癌裸鼠移植瘤的实验研究

目的:研究Angiostatin基因治疗对人肝癌裸鼠皮下移植瘤的抑制作用及其相关机理。方法:使用人原发性肝癌细胞株SMMC-7721建立人肝癌裸鼠皮下移植瘤动物模型,质粒用脂质体DOTAP介导转染细胞。将荷瘤裸鼠随机分为两组,分别注射质粒PcDNA3、Angiostatin/PcDNA3,观察两组动物的肿瘤生长曲线,检测肿瘤的Angiostatin、VEGF、HIF-1α表达和微血管密度(MVD),利用TUNEL染色法行原位细胞凋亡分析。结果:Angiostatin基因治疗在早期具有抑制肿瘤生长的作用,大约1周后肿瘤以更快的速度生长并迅速赶上空质粒对照组肿瘤;Angiostatin基因治疗组的肿瘤组织中有An-giostatin的局部高表达,MVD(24.8±2.8)低于空质粒对照组(30.2±4.1)(P〈0.05)。肿瘤组织中HIF-1α蛋白局部高表达,VEGF表达高于空质粒对照组,细胞凋亡指数(2.87±0.48)高于空质粒对照组(1.55±0.43)(P〈0.01)。结论:Angiostatin基因治疗对人肝癌裸鼠皮下移植瘤的生长具有一定的抑制作用,肿瘤对Angiostatin基因治疗可以产生耐受性。     

嘌呤核苷酸对海洛因成瘾大鼠垂体FSH、LH基因表达及内分泌细胞超微结构的影响

目的:研究嘌呤核苷酸对海洛因依赖及戒断大鼠腺垂体卵泡刺激素(FSH)、黄体生成素(LH)/间质细胞刺激素(ICSH)基因表达情况及远侧部生长激素细胞和促性腺激素细胞超微结构的影响。方法:采用逐渐递增剂量法腹腔注射海洛因,建立海洛因依赖及戒断大鼠模型,92只Wistar大鼠随机分成对照组(生理盐水)、核苷酸组(不给海洛因)、海洛因组、海洛因+核苷酸组(同时给药)、戒断3 d组、戒断9 d组、核苷酸3 d组(海洛因停药后给核苷酸3 d)及核苷酸9 d组(海洛因停药后给核苷酸9 d),RT-PCR法半定量分析各组大鼠垂体FSH及LHmRNA表达的变化,电镜观察腺垂体远侧部生长激素细胞和促性腺激素细胞超微结构的变化。结果:与对照组大鼠垂体FSH mRNA的相对表达量(0.045±0.009)比较,核苷酸组(0.099±0.018)、海洛因+核苷酸组(0.177±0.046)、核苷酸3 d组(0.151±0.030)和核苷酸9 d组(0.184±0.028)明显升高(P<0.01);与对照组大鼠垂体LHmRNA的相对表达量(0.700±0.099)比较,海洛因+核苷酸组(0.950±0.169)、核苷酸3 d组(0.990±0.171)和核苷酸9 d组(0.960±0.147)明显升高(P<0.01)。与对照组相比,海洛因组腺垂体远侧部生长激素细胞和促性腺激素细胞核呈不规则形,核膜不清,核基质略致密,核内异染色质趋边、凝集。胞质内粗面内质网扩张,线粒体肿胀、嵴断裂且呈空泡化,分泌颗粒明显减少,部分细胞内可见髓样小体。海洛因+核苷酸组生长激素细胞和促性腺激素细胞与对照组相比无明显变化。结论:短期应用海洛因对大鼠垂体FSH和LH的基因表达影响不明显,与海洛因同时给予嘌呤核苷酸、或海洛因撤药后给予嘌呤核苷酸则明显促进了FSH、LH基因的表达。海洛因可造成大鼠腺垂体远侧部生长激素细胞和促性腺激素细胞超微结构的损伤;嘌呤核苷酸能一定程度地减轻或抑制海洛因对腺垂体远侧部生长激素细胞和促性腺激素细胞超微结构的损害。     

原代培养和纯化人垂体腺瘤细胞的研究

垂体腺瘤约占颅内肿瘤的10%～15%,而其中侵袭性垂体腺瘤又占垂体腺瘤的5%～85%.相对一般垂体腺瘤而言,侵袭性垂体瘤常侵犯海绵窦、蝶窦、鞍上、斜坡等周围结构,无法做到彻底切除,且易复发[1].我们对人垂体腺瘤细胞进行了体外原代培养,利用无血清培养技术得到了纯化的垂体腺瘤细胞.     

侵袭性垂体腺瘤成纤维细胞的体外培养及其生物学特性

目的建立人侵袭性垂体腺瘤成纤维细胞分离培养的方法，探讨其生物学特性及其在垂体瘤侵袭性生长中的作用。方法差速贴壁筛选法分离培养侵袭性垂体腺瘤成纤维细胞，进行原代及传代培养，倒置相差显微镜下观察原代及传代细胞形态及生长特性，免疫组化染色进行成纤维细胞鉴定，MTT法绘制传代细胞的生长曲线，电镜观察其超微结构。结果侵袭性垂体腺瘤成纤维细胞体外培养生长状况良好，具有活跃的增殖能力。细胞胞体较大，形态不规则，细胞质中可见丰富的粗面内质网和核糖体，高尔基复合体发达，Ⅰ型胶原及波形蛋白表达率在95％以上。结论应用差速贴壁筛选法成功培养出侵袭垂体腺瘤成纤维细胞，其具有旺盛的增殖能力，可能在垂体瘤侵袭性生长过程中起重要作用。     

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.