结缔组织生长因子在血吸虫病肝纤维化鼠肝窦内皮细胞中的表达及意义

目的:动态观察结缔组织生长因子(CTGF)蛋白在血吸虫病肝纤维化鼠肝窦内皮细胞表达水平的时相变化,探讨其与肝窦内皮细胞下基底膜形成的关系。方法:采用腹部敷贴法建立血吸虫病肝纤维化模型,HE、Masson染色和透射电镜观察其病理变化;免疫组化技术检测CTGF、Ⅳ型胶原(ColⅣ)和层粘连蛋白(LN)在小鼠肝脏组织中的表达和分布;并应用彩色图像分析仪进行定量分析。结果:与正常对照组比较,模型组鼠肝窦内皮细胞表达CTGF蛋白阳性或强阳性,肝窦壁LN、ColⅣ表达水平增高,且随着肝纤维化的发展,CTGF和LN、ColⅣ表达逐渐增强,肝窦内皮下基底膜逐渐增厚。图像定量分析两组平均吸光度值、平均灰度值和阳性面积比具有统计学差异;CTGF蛋白与LN、ColⅣ水平呈正相关。结论:血吸虫病肝纤维化时小鼠肝窦内皮细胞通过CTGF蛋白表达上调,调控细胞外基质产生,导致ColⅣ、LN分泌增加,参与肝窦内皮下基底膜形成,从而引起肝内微循环障碍。     

Molecular analysis of C3 allotypes in patients with nephritic factor.

The autoantibody nephritic factor (NeF) leads to complement consumption in vivo and is associated with type II mesangiocapillary glomerulonephritis (MCGN II) and partial lipodystrophy (PLD). The third component of complement (C3) exists in two common allotypic forms, C3S and C3F, distinguished at the protein level by electrophoresis. An increased frequency of the rarer C3F allele has been reported in several autoimmune conditions, including one small series of patients with NeF. However, patients with NeF have low levels of circulating C3 so that allotyping at the protein level is difficult. The molecular basis of the S/F polymorphism has recently been established: a single base change at the DNA level encodes a single amino acid substitution at the protein level. A second polymorphism, closely linked to the first, is defined by the MoAb HAV 4-1, and is also due to a single base change. These polymorphisms can therefore be analysed at the DNA level. We have used the amplification refractory mutation system (ARMS), a modification of the polymerase chain reaction (PCR), to analyse these two C3 polymorphisms at the DNA level in 26 patients with NeF. The allele frequencies of C3S and C3F were 0.673 and 0.327 (predicted values 0.79 and 0.2, chi 2 = 4.813, P < 0.05), giving a relative risk of 2.1 for the development of NeF conferred by the presence of a C3F allele. The HAV 4-1 allele frequencies were (-) 0.71 and (+) 0.29, i.e. not significantly different than predicted from the linked C3S/F allele frequencies. This is the largest series of patients with NeF yet published, and our data confirm an association between C3F and NeF. Possible mechanisms for for this link are discussed.     

HIV/AIDS人员心理卫生状况与心理社会影响因素

目的 :研究HIV/AIDS (艾滋病毒感染者 /艾滋病患者 )人群的心理卫生状态与心理社会因素的影响。方法 :采用病例对照研究方法。病例组与对照组两组对象均接受一般情况问卷、症状自评量表(Symptomchecklist 90 ,SCL -90 )、生活事件量表 (LifeEventScale ,LES)、生活质量综合评定问卷 (GenericQualityofLifeInventory -74,GQOLI -74)、自我接纳问卷 (Self -AcceptanceQuestionnaire ,SAQ )五个量表的测查。结果 :(1)病例组的SCL -90各个因子分高于对照组 (P <0 0 0 1) ;(2 )病例组总生活事件和负性生活事件得分高于对照组 (P =0 0 0 1) ;(3 )病例组自我接纳得分低于对照组 (P =0 0 0 1) ;(4 )病例组生活质量综合评定得分低于对照组 (P <0 0 0 1) ;(5 )多因素逐步回归分析 ,SCL -90总因子分相关因素主要为躯体症状严重程度 (负向评分 )、负性生活事件和自我接纳因子 ,躯体症状严重程度与SCL -90总因子分呈负相关 ,负性生活事件得分与SCL -90总因子分呈正相关 ,自我接纳得分与SCL -90总因子分呈负相关。结论 :HIV/AIDS组较对照组心理健康状态有明显差异 ,有较多的负性生活事件 ;自我接纳得分低于对照组 ;生活质量低于对照组。     

Factors related to postmenopausal muscle performance: a cross-sectional population-based study

The aim of the present study was to investigate cross-sectionally the association of postmenopausal muscle strength with simple performance tests. A random sample of 1,166 naturally postmenopausal women (born 1932–1941) was selected from the Kuopio Osteoporosis Risk Factor and Prevention (OSTPRE) study cohort. Grip and quadriceps strength were measured with strain gauge dynamometers and reported in both absolute values (KPa and kg) and per kilogram of body mass (N cm^–2 kg^–1 and N kg^–1). In addition, two performance tests, ability to stand on one foot and ability to squat down to touch the floor were carried out. A five-category self-assessment of overall health (very good, good, moderate, bad, and very bad) was obtained by postal questionnaire. The women that were able to stand on one foot and able to squat down to touch the floor had greater grip and quadriceps strength than their counterparts (P<0.001 and P<0.03 in ANOVA, respectively). In addition, self-assessed health had a strong positive association with muscle strength in the grip and leg extensor muscles in ANOVA (P<0.001 between very good and moderate or poorer state of health) and regression model (P<0.001). Adjustment for age, duration of menopause, body mass, height, physical activity level, use of HRT, and number of diseases and medications did not change any of the main effects. Also, there were no differences in results between absolute measurement values and values reported per kilogram of body mass. According to the present study, a simple performance test may be useful in the prediction of postmenopausal muscle strength. Furthermore, self-assessed state of health is strongly associated with muscle strength in postmenopausal women.     

Endothelial nitric oxide synthase gene polymorphism in women with idiopathic recurrent miscarriage

BACKGROUND: Lack of endothelium-derived nitric oxide is associated with vasospasm and vascular infarction. We investigated the relationship between idiopathic recurrent miscarriage and a polymorphism of the gene encoding endothelial nitric oxide synthase (NOS3). METHOD: In a prospective case-control study, 105 women with idiopathic recurrent miscarriage and 91 healthy controls were investigated. We used the polymerase chain reaction to identify the different alleles of a 27 base pair tandem repeat polymorphism in intron 4 of the NOS3 gene. RESULTS: The wild type B allele was identified on 329 out of 392 chromosomes (frequency 0.84). The polymorphic A allele was present on 63 chromosomes (frequency 0.16). The genotype frequencies were as follows: 68% (B/B), 31% (A/B) and.5% (A/A). The distribution of genotype frequencies was significantly different between the study and control groups for allele A/B heterozygotes (NOS3(A/B)) (36.7 versus 23.8%, P = 0.03, OR 1.6, 95% CI 1.1--3.8). Only one individual was homozygous for the A allele (NOS3(A/A)). She was in the study group. Between women with primary and secondary recurrent miscarriages, no statistically significant difference between the distribution of NOS3(A/B) genotypes (28 versus 34%) was observed. CONCLUSIONS: These data support a role for the NOS3 gene as a genetic determinant of the risk of idiopathic recurrent miscarriage.     

Molecular basis of congenital insensitivity to pain with anhidrosis (CIPA): Mutations and polymorphisms in TRKA (NTRK1) gene encoding the receptor tyrosine kinase for nerve growth factor

Congenital insensitivity to pain with anhidrosis (CIPA), also referred to as hereditary sensory and autonomic neuropathy type IV (HSAN‐IV), is an autosomal recessive hereditary disorder characterized by recurrent episodic fever, anhidrosis (inability to sweat), absence of reaction to noxious stimuli, self‐mutilating behavior, and mental retardation. The TRKA (NTRK1) gene located on chromosome 1 (1q21‐q22), consists of 17 exons and spans at least 23 kb. TRKA encodes the receptor tyrosine kinase (RTK) for nerve growth factor (NGF) and is the gene responsible for CIPA. Defects in NGF signal transduction at the TRKA receptor lead to failure to support survival of sympathetic ganglion neurons and nociceptive sensory neurons derived from the neural crest. Thirty‐seven different TRKA mutations, identified in patients in various countries, including nine frameshift, seven nonsense, seven splice, and 14 missense mutations, are distributed in an extracellular domain involved in NGF binding, as well as in the intracellular signal‐transduction domain. Extensive analysis of CIPA mutations and associated intragenic polymorphisms should facilitate detection of CIPA mutations and aid in the diagnosis and genetic counseling of this painless but severe genetic disorder with devastating complications. In addition, naturally occurring TRKA missense mutations with loss of function provide considerable insight into the structure–function relationship in the RTK family. Further, molecular pathology of CIPA would provide unique opportunities to explore critical roles of the autonomic sympathetic nervous system as well as peripheral sensory nervous system that transmit noxious stimuli in humans. Hum Mutat 18:462–471, 2001. © 2001 Wiley‐Liss, Inc.     

人血浆高密度脂蛋白对大鼠内毒素血症的治疗及防护作用研究

目的:观察人血浆高密度脂蛋白(HDL)对大鼠内毒素血症的治疗和防护效果。方法:采用鲎试剂法和放射免疫分析法于3个时点动态测定对照组、治疗组和防护组大鼠血浆内毒素(ET)水平和肿瘤坏死因子(TNF)浓度并观察血压及存活时间。结果:①输注ET后对照组大鼠血压进行性下降(P<0.01);治疗组大鼠输注HDL后其血压虽也降低但下降程度明显弱于对照组(P<0.01);防护组大鼠在输注ET后血压无明显下降(P>0.05)。治疗组及防护组大鼠存活时间均明显长于对照组(P<0.01)。②3组大鼠血浆ET水平在各时点均无明显变化(P>0.05)。③治疗组及防护组大鼠血浆TNFα水平于第3时点均明显降低(P>0.05)。结论:人血浆HDL能减轻或抑制内毒素血症大鼠血压下降并明显延长其存活时间,对内毒素血症具有良好的治疗和防护作用,此作用可能与其抑制TNF释放有关。     

Effects of nerve growth factor and glucocorticoid on cultured human pheochromocytoma cells

Primary cell cultures of two human pheochromocytomas (PC) that were associated with high serum levels of adrenaline and noradrenaline were developed to study the effects of nerve growth factor (NGF) and dexamethasone on the morphology and function of PC cells in vitro. By phase-contrast microscopy, cultured cells were small and hyperchromatic on the first day of culture; neurite-like processes that extended to other cells developed several days later and were maintained for more than 3 months. NGF (100ng/ml), dexamethasone (10^–5M), or NGF + dexamethasone were added to the culture media 2 weeks after the cultured cells had stabilized. Catecholamine concentrations in the medium were maintained at higher levels after addition of NGF, dexamethasone, or NGF + dexamethasone as compared to control cells. In the presence of NGF, extension of neurite-like processes was clearly accelerated, while high levels of dexamethasone inhibited growth of processes. These in vitro studies showed that the addition of NGF or the removal of dexamethasone induces differentiation of adrenal neurons present in pheochromocytomas, suggesting that adrenocortical steroid hormones influence the morphological control of adrenal medullary cells.     

机械牵张对大鼠心室肌蛋白激酶B磷酸化及心钠素分泌的影响

目的:研究机械牵张对大鼠心肌蛋白激酶B(Akt)活化和心钠素(ANF)分泌的影响。方法:采用Langendorff方法灌流大鼠心脏,膨胀球囊持续牵张左心室,从左心室游离心肌,提取胞浆蛋白,用Western blot检测磷酸化Akt、总Akt水平;收集冠脉流出液,用放射免疫分析法检测冠脉流出液中ANF含量。结果:持续牵张不影响灌流心脏的心率和冠脉流出量。但经过20min持续牵张,牵张组心脏灌流液中ANF含量(209.89±65.45pg/ml)较对照组(108.84±25.18pg/ml)明显增高(P<0.01);牵张组大鼠左心室心肌组织磷酸化Akt水平(0.76±0.03)明显高于对照组(0.32±0.02),而总Akt水平与对照组相比无显著性差异。结论:机械牵张可引起心脏心钠素的分泌增加,其机制可能与胞内Akt信号通路的激活有关。     

HOXB7 siRNA表达载体在裸鼠体内对人恶性黑色素瘤细胞株的影响

目的 探讨转染同源盒第7基因(HOXB7)siRNA质粒表达载体对人恶性黑色素瘤细胞株A375在裸鼠体内生长的影响.方法 裸鼠皮下接种人恶性黑色素瘤A375细胞,对2周后形成的瘤块进行分组干预.随机分为生理盐水对照组、阴性质粒组、HOXB7质粒组,观察转染后各组裸鼠移植瘤的生长情况;免疫组化法比较瘤体内微血管密度(MVD).结果 HOXB7质粒组的裸鼠移植瘤块生长慢、体积明显小于生理盐水对照组[(0.134±0.039)cm3比(1.006±0.235)cm3,P＜0.05],而阴性质粒组瘤块体积大小与生理盐水对照组无统计学差异[(0.929±0.157)cm3比(1.006±0.235)cm3,P＞0.05].HOXB7质粒组裸鼠体内肿瘤MVD低于生理盐水对照组[(2.8±1.9)比(19.9±5.6),P＜0.05],阴性质粒组与生理盐水对照组无统计学差异[(18.1±5.5)比(19.9±5.6),P＞0.05].结论 针对HOXB7 siRNA质粒表达载体可有效抑制A375细胞裸鼠体内肿瘤生长和瘤内血管生成.