构建肠系膜微淋巴管内皮细胞培养技术并观察休克淋巴液环境中内皮细胞的形态变化

目的:建立肠系膜微淋巴管内皮细胞的培养技术,观察不同体积分数的休克淋巴液对大鼠肠系膜微淋巴管内皮细胞形态的影响。方法:实验于2004-02/07在河北北方学院病理生理学教研室及实验中心细胞培养室完成。选择健康雄性Wistar大鼠,体质量分别为220～300g和50～80g。实验方法:①无菌条件下制备大鼠重症失血性休克模型,引流肠系膜淋巴液或收集门静脉血;同时另取大鼠,引流正常淋巴液或正常门静脉血。②从动物种类、培养基、植块方法、胎牛血清浓度等方面对植块培养法进行改良,进行肠系膜微淋巴管内皮细胞原代培养并传代。实验分组:分为6组:胎牛血清组:培养液为DMEM 体积分数为0.1的胎牛血清;正常淋巴液组:培养液为DMEM 正常淋巴液;休克淋巴液组:培养液为DMEM 体积分数为0.04,0.06,0.08,0.10的休克淋巴液;正常血浆组:培养液为DMEM 正常血浆;休克血浆组:培养液为DMEM 休克血浆;无血清对照组:培养液为DMEM。实验评估:①光镜下观察大鼠肠系膜微淋巴管内皮细胞原代培养及传代情况。②光镜、扫描电镜及透射电镜下观察不同体积分数的休克淋巴液及不同作用时间(4,8,12h)对肠系膜微淋巴管内皮细胞形态及超微结构的影响。结果:①光镜下肠系膜微淋巴管内皮细胞原代培养及传代情况:内皮细胞呈扁平的梭形或多边形,大小均匀,胞核清晰,呈卵圆形。细胞生长融合形成单层后,呈典型的鹅卵石或铺路石样镶嵌状排列生长。②光镜下在休克淋巴液中肠系膜微淋巴管内皮细胞形态:体积分数为0.04的休克淋巴液作用4h时细胞逐渐收缩、变圆直至脱落漂浮,随着休克淋巴液体积分数增加及作用时间延长,细胞损伤逐渐加重,体积分数为0.08的休克淋巴液作用4h时核旁出现空泡,体积分数为0.10的休克淋巴液作用12h时细胞裂解成碎片。其他各组作用12h肠系膜微淋巴管内皮细胞形态无显著改变。③扫描电镜下在休克淋巴液中肠系膜微淋巴管内皮细胞形态:体积分数为0.04的休克淋巴液作用4h部分细胞逐渐收缩离壁,细胞间隙增大、细胞边缘的突起拉长、缩短、断裂,作用8,12h可见凋亡小体。其他各组作用12h肠系膜微淋巴管内皮细胞形态无显著改变。④透射电镜下在休克淋巴液中肠系膜微淋巴管内皮细胞形态:体积分数为0.04的休克淋巴液作用4h细胞膜形态不规整,胞浆内质网等膜性细胞器扩张,核形态不规则,作用8h线粒体呈球形扩张,细胞核逐渐出现固缩、似细胞凋亡改变,核周腔变宽;体积分数为0.08的休克淋巴液作用时间8h细胞内出现囊泡,内质网不同程度扩张,线粒体主要表现为浓缩、深染等改变,细胞膜边界不清,出现起泡、破碎等现象。其他各组作用12h肠系膜微淋巴管内皮细胞形态无显著改变。结论:成功建立了肠系膜微淋巴管内皮细胞的培养技术;休克淋巴液可导致肠系膜微淋巴管内皮细胞形态及超微结构损伤。     

辽西地区农村汉族青少年体型发育规律的Heath-Carter体型法分析

目的:分析辽西地区农村汉族青少年的体型发育规律和特点,为体质人类学补充必要的数据。对象:按整群分层抽样法,抽取2001-07/2003-09辽西地区锦州、盘锦、葫芦岛、义县和绥中的农村13所中小学7～19岁经学校正常体质检查证明身体健康的汉族学生为调查对象,按性别分两大组,每大组按年龄分12小组,7～18岁每岁为1个年龄组,18～19岁为1个年龄组,每小组44～82人,共分24组,搜集完整资料1280名(男666名,女614名)。采用Heath-Carter体型法,每项指标测量2次,取平均值,10项指标由专人负责,测试数据按年龄和性别在微机中建立数据库,依次计算出各年龄组的内因子、中因子和外因子,体型图上的X,Y坐标值,身高/体质量1/3,样本中平均体型点到所有体型点空间距离的均数,三维空间中两个体型点间的差异,体脂百分比和各类体型分布频数。结果:参加调查1280名,均进入结果分析。①7～17岁青少年身高、体质量随年龄的增加而增长。平均身高、体质量男生大于女生。身高/体质量1/3指数除7、8、10、12和13岁外,各年龄组男生>女生,平均值男生>女生。体脂含量除8岁外,其他年龄组女生>男生,平均值女生>男生。②男生的体型均值3.6-2.5-3.7,属外胚层-内胚层均衡体型,女生平均体型值为4.3-2.2-3.3,属于偏外胚层的内胚层体型。体型频数的变化提示辽西农村汉族男生体型分布在内胚层和外胚层的体型较多,女生分布较集中,主要在内胚层体型。③内因子男生在3.0～4.0,女生在3.5～5.2,各年龄组女生>男生;中因子男生在1.6～3.9,女生在0.8～3.4,除7、11和17岁女生>男生外,其他年龄组男生>女生或男女相等,平均值男生>女生。外因子男生在3.0～5.0,女生在3.0～4.1,7、8、10、12、13岁女生>男生,11、14、15、16、17、18岁男生>女生。因此,女生的内因子占优势,皮下脂肪更发达,体态丰满,男生中因子占优势,骨骼粗壮,肌肉发达,随着年龄的增长,男生的身体相对瘦高程度增长,身材修长。④男女各年龄组间体型比较,7,8,9,10,11,12,13,14,15,16,17,18～19岁三维空间中两个体型点间的差异值分别为0.45,0.67,1.03,0.56,0.76,0.30,0.93,2.60,1.86,1.37,1.21,1.47,8～11岁年龄组男女间体型差异有显著性(t=2.66,2.75,2.95,3.58,P<0.05)。⑤与城市青少年相比,农村青少年身高低5～6cm(男女分别为u=6.24,5.90,P<0.01),体质量低3～6kg(男女分别为u=6.93,4.60,P<0.01)。结论:辽西地区农村男生较女生骨骼粗壮,肌肉发达,女生的皮下脂肪更发达,体态丰满,随着年龄的增长,男生的身体相对瘦高,身材修长。与城市青少年相比,农村青少年仍矮瘦,与国外资料和国内少数民族相比,辽西地区农村汉族青少年骨骼肌肉欠发达,身材修长。     

Prognostic discrimination in "good-risk" chronic granulocytic leukemia

The prognostic significance of disease features recorded at the time of diagnosis was examined among 813 patients with Philadelphia chromosome- positive, nonblastic chronic granulocytic leukemia (CGL) collected from six European and American series. The survival pattern for this population was typical of "good-risk" patients, and median survival was 47 mo. There were multiple interrelationships among different disease features, which led to highly significant correlations with survival for some that had no primary prognostic significance, such as hematocrit. Multivariable regression analysis indicated that spleen size and the percentage of circulating blasts were the most important prognostic indicators. These features, and age, behaved as continuous variables with progressively unfavorable import at higher values. The platelet count did not influence survival significantly at values below 700 X 10(9)/liter but was increasingly unfavorable above this level. Basophils plus eosinophils over 15%, more than 5% marrow blasts, and karyotypic abnormalities in addition to the Ph1 were also significant unfavorable signs. The Cox model, generated with four variables representing percent blasts, spleen size, platelet count, and age, provided a useful representation of risk status in this population, with good fit between predicted and observed survival over more than a twofold survival range. A hazard function derived from half of the patient population successfully segregated the remainder into three groups with significantly different survival patterns. We conclude that it should be possible to identify a lower risk group of patients with a 2-yr survival of 90%, subsequent risk averaging somewhat less than 20%/yr and median survival of 5 yr, an intermediate group, and a high- risk group with a 2-yr survival of 65%, followed by a death rate of about 35%/yr and median survival of 2.5 yr.     

Inhibiting glycogen synthase kinase-3 reduces endotoxaemic acute renal failure by down-regulating inflammation and renal cell apoptosis

Background and purpose:

Excessive inflammation and apoptosis are pathological features of endotoxaemic acute renal failure. Activation of glycogen synthase kinase-3 (GSK-3) is involved in inflammation and apoptosis. We investigated the effects of inhibiting GSK-3 on lipopolysaccharide (LPS)-induced acute renal failure, nuclear factor-κB (NF-κB), inflammation and apoptosis.

Experimental approach:

The effects of inhibiting GSK-3 with inhibitors, including lithium chloride (LiCl) and 6-bromo-indirubin-3′-oxime (BIO), on LPS-treated (15 mg·kg⁻¹) C3H/HeN mice (LiCl, 40 mg·kg⁻¹ and BIO, 2 mg·kg⁻¹) and LPS-treated (1 µg·mL⁻¹) renal epithelial cells (LiCl, 20 mM and BIO, 5 µM) were studied. Mouse survival was monitored and renal function was analysed by histological and serological examination. Cytokine and chemokine production, and cell apoptosis were measured by enzyme-linked immunosorbent assay and terminal deoxynucleotidyl transferase-mediated dUTP–biotin nick-end labelling staining, respectively. Activation of NF-κB and GSK-3 was determined by immunostaining and Western blotting, respectively.

Key results:

Mice treated with GSK-3 inhibitors showed decreased mortality, renal tubular dilatation, vacuolization and sloughing, blood urea nitrogen, creatinine and renal cell apoptosis in response to endotoxaemia. Inhibiting GSK-3 reduced LPS-induced tumour necrosis factor-α (TNF-α) and CCL5/RANTES (released upon activation of normal T-cells) in vivo in mice and in vitro in murine kidney cortical collecting duct epithelial M1 cells. Inhibiting GSK-3 did not block TNF-α-induced cytotoxicity in rat kidney proximal tubular epithelial NRK52E or in M1 cells.

Conclusions and implications:

These results suggest that GSK-3 inhibition protects against endotoxaemic acute renal failure mainly by down-regulating pro-inflammatory TNF-α and RANTES.     

Perioperative management and the associated rate of adverse events in dermatological procedures performed by dermatologists in New Zealand

Dermatological surgery performed in an outpatient setting is common and generally perceived as safe, but the potential for serious adverse events does exist. Furthermore, there is a current lack of guidelines regarding preoperative and intraoperative monitoring of such patients. This is a retrospective study that involved a written questionnaire sent to current practising New Zealand dermatologists. Aspects investigated include their practice relating to preoperative assessments and intraoperative monitoring during standard dermatological procedures, and the resulting rate of adverse events. We found that most respondents performed dermatological procedures in dedicated theatres in outpatient clinics. The majority of survey respondents would screen for and optimize risk factors prior to surgery. Most respondents would not record vital sign measurements either preoperatively or intraoperatively. Antibiotic prophylaxis was generally only prescribed if clinically indicated, and anticoagulation and antiplatelet therapies were in the majority of cases never withheld prior to surgery. Infection (<3.5%) and bleeding (<2%) were the most common postoperative complications, with other serious adverse events being extremely rare. Although dermatological surgery continues to be safely performed in the outpatient setting, attempts should still be made to identify patients who are at higher risk for surgery and extra precautions should be applied to these selected patients.     

Asynchronism of the recovery of baroreflex sensitivity, blood pressure, and consciousness from anesthesia in rats

Anesthesia inhibits arterial baroreflex functions such as baroreflex sensitivity （BRS）. The main objective of the present study was to determine the time course of BRS recovery from anesthesia and to determine whether BRS recovery is synchronous with the recovery of consciousness and blood pressure （BP）. Experiments were performed in male Sprague-Dawley rats using different commonly used anesthetics at routine doses through intraperitoneal administration： （1） diazepam/ketamine,     

Synergism of atenoloi and nitrendipine on hemodynamic amelioration and organ protection in hypertensive rats

OBJECTIVE： This study was designed to investigate the possible synergism of atenolol and nitrendipine on blood pressure （BP） and blood pressure variability （BPV） reductions, baroreflex sensitivity （BRS） amelioration, and organ protection in hypertensive rats. METHOD： The dose was 20 mg/kg for atenolol, 10 mg/kg for nitrendipine and 20 ＋ 10 mg/kg for the combination of these two drugs. In an acute study, a single dose was given via a catheter previously inserted into the stomach in spontaneously hypertensive rats （SHR）.     

Effect of respiratory virus infections including rhinovirus on clinical status in cystic fibrosis

One hundred and eight patients with cystic fibrosis were investigated over one year to determine whether an association existed between rhinovirus or other respiratory virus infection and clinical status. Forced expiratory volume in one second (FEV1), forced vital capacity (FVC), Shwachman score, Chrispin-Norman chest radiograph score, and percentage weight for height were recorded at the beginning and end of the study; days of intravenous antibiotics were noted. Nasopharyngeal aspirates were taken for viral studies during respiratory exacerbations. Serum was collected at enrollment and 2-6 weeks after each respiratory exacerbation. One hundred and fifty seven exacerbations occurred in 76 patients. Respiratory virus infection was detected in 44 exacerbations and rhinovirus was present in 16% (25/157) of exacerbations. Patients with one or more respiratory virus infections were compared with those who had none. When all respiratory virus infections were considered, patients had a significantly greater deterioration in Shwachman score and received significantly more days of intravenous antibiotics. When rhinovirus was considered separately, patients received significantly more days of intravenous antibiotics, but showed no deterioration in clinical status. However, patients infected with another respiratory virus had a significant decline in FEV1, with trends towards significance for decline in FVC and Shwachman score.     

Living high–training low altitude training: effects on mucosal immunity

Secretory immunoglobulin A (sIgA) is the major immunoglobulin of the mucosal immune system. Whereas the suppressive effect of heavy training on mucosal immunity is well documented, little is known regarding the influence of hypoxia exposure on sIgA during altitude training. This investigation examined the impact of an 18-day Living high–training low (LHTL) training camp on sIgA levels in 11 (six females and five males) elite cross-country skiers. Subjects from the control group (n=5) trained and lived at 1,200 m of altitude, whereas, subjects from the LHTL group (n=6) trained at 1,200 m, but lived at a simulated altitude of 2,500, 3,000 and 3,500 m (3×6-day, 11 h day^–1) in hypoxic rooms. Saliva samples were collected before, after each 6-day phases and 2 weeks thereafter (POST). Salivary sIgA, protein and cortisol were measured. There was a downward trend in sIgA concentrations over the study, which reached significance in LHTL (P<0.01), but not in control (P=0.08). Salivary IgA concentrations were still lower baseline at POST (P<0.05). Protein concentration increased in LHTL (P<0.05) and was negatively correlated with sIgA concentration after the 3,000 and 3,500 m-phase and at POST (P<0.05 all). Cortisol concentrations were unchanged over the study and no relationship was found between cortisol and sIgA. In summary, data were strongly suggestive of a cumulative negative effect of physical exercise and hypoxia on sIgA levels during LHTL training. Two weeks of active recovery did not allow for proper sIgA recovery. The mechanism underlying this depression of sIgA could be mediated by neural factors.