氟中毒病区家猪骨胶原形态学观察

骨胶原是骨组织的基本支架,也是氟作用的靶组织,氟可减少胶原的合成,并产生异常胶原,使其完整性下降,胶原纤维是折光性物质,在正交偏振光(偏光)显微镜下可清晰地观察到成熟胶原纤维构成的组织形态,如胶原纤     

过量氟与软骨代谢

氟中毒与软骨组织代谢有着密切的联系，过量氟对软骨有明显的损害作用，可造成钙、磷等无机元素代谢异带．导致软骨蛋白多糖合成与分解失调，干扰软骨基质胶原代谢及改变软骨组织酶的活性。     

氟中毒动物模型胶原免疫组化及Ⅰ胶原MRNA的测定

目的通过研究氟中毒动物模型骨Ⅰ型胶原免疫组化及Ⅰ型胶原MRNA的影响,为预防和治疗氟骨症提供基础理论。方法对正常与模型组大鼠股骨以图像分析系统观察Ⅰ型胶原免疫组化及Ⅰ型胶原MRNA原位杂交检测。结果得出了各组试样的阳性染色分别面积、平均灰度值、阳性反应面积等指标。结论模型组各项指标显著低于正常对照组（P〈0.05）,氟中毒动物模型胶原免疫组化及Ⅰ型胶原MRNA指标发生改变。     

胶原酶抑制因子在氟中毒发病学中意义的研究

我们使用胶原酶抑制因子单克隆抗体和酶免疫夹心法测定了氟中毒病区儿童血清胶原酶抑制因子和胶原代谢产物羟脯氨酸的含量。发现氟中毒病区儿童血清胶原酶抑制因子和尿中羟脯氨酸含量明显高于正常对照组(P<0.01)。胶原酶抑制因子在调节胶原酶活性中起着重要作用,特别是控制结缔组织的降解。“氟中毒的主要靶组织是胶原蛋白”,测定胶原酶抑制因子和羟脯氨酸含量对氟中毒的早期生化诊断和认识其发病机理有着重要意义。     

氟砷联合中毒患者无机元素和胶原代谢状况观察

目的观察改水后地方性氟中毒、氟砷联合中毒患者无机元素和胶原代谢的改变.方法用氯胺T法测定尿羟脯氨酸含量;用氟离子选择电极法测定尿氟含量;用二乙氨基二硫代甲酸银法测定尿砷含量;用Baird PS-4型等离子体光谱仪测定尿中钾、钠、钙、磷、镁、锌含量;用美国BECKMAN公司LX 20型全自动生化分析仪检测血清中钙、磷、镁、锌、铁含量.结果各组之间尿氟、砷、HOP/Cr、钠、钾和血清钙、镁含量的差别均有统计学意义(P<0.05).结论单纯氟中毒和氟砷联合中毒所致人体的无机元素和胶原代谢紊乱在脱离高氟高砷暴露20年后不能完全纠正,氟砷联合中毒患者体内无机元素和胶原代谢紊乱的恢复比单纯氟中毒患者更缓慢.     

硼、硒、氟宁、维生素C对氟中毒动物生化指标的影响

通过给大白鼠饮用高氟水,导致骨软化型氟中毒动物模型,同时分别给予四硼酸钠、亚硒酸钠、蛇纹石、维生素C,观察动物血中钙、磷含量及碱性磷酸酶活性,骨中胶原、钙及氟水平和尿氟含量的变化。 实验结果显示:硼能降低血清ALP活性;氟宁能增加骨中胶原及钙含量;硒能提高血钙浓度加速尿氟排泄;维生素C亦能增加血钙、骨胶原和尿氟含量。这些药物对氟的生化改变的影响,对防治氟中毒将起重要作用。     

氟中毒大鼠血清皮质酮含量的变化

本文观察了氟中毒大鼠血清皮质酮含量变化。发现在不同的实验条件下,氟中毒大鼠血清皮质酮含量均明显降低。认为在氟,糖皮质激素以及骨和胶原代谢之间一定存在某种联系,而这种联系可能在氟中毒的发病机制中占重要地位。     

大鼠慢性氟中毒神经细胞病理改变的研究

目的 研究慢性氟中毒对神经细胞的作用机制及病理改变。方法 利用光电镜观察了慢性氟中毒大鼠脑组织的病理学改变。结果 慢性氟中毒大鼠组织光镜下的病理改变主要表现在海马CA4区,以神经细胞核浓度、固缩、顶树突明显拉长为特点;电镜下的病理改变与光镜的改变一致。结论 慢性氟中毒鼠脑组织的病理改变具有选择性易损的特点：最易受损的部位是海马CA4区,与大鼠前脑缺血再灌流模型脑细胞病理改变相似。     

氟中毒对大鼠肝肾组织磷脂脂肪酸组成的影响

目的　研究氟中毒对大鼠肝、肾组织磷脂脂肪酸组成的影响。方法　选用 Wistar纯系大鼠 ,饲以高浓度含氟水 1个月 ,来复制慢性氟中毒动物模型。用改良 Falch方法提取和分离动物肝、肾组织磷脂 ,用气相色谱法分离和测定磷脂各类脂肪酸构成。结果　所有饲以高浓度氟水的动物均有不同程度的氟斑牙形成 ,尿氟含量增高 ,肝和肾组织蛋白质含量降低等慢性氟中毒的表现 ;氟中毒动物肝、肾组织磷脂脂肪酸组成发生异常改变 ,表现为多不饱和脂肪酸减少 ,饱和脂肪酸组成增加 ,减少的多不饱和脂肪酸种类是花生四稀酸和二十二碳六烯酸。结论　表明长期摄入过量氟可导致肝、肾组织磷脂脂肪酸组成改变 ,磷脂多不饱和脂肪酸减少可能与自由基水平升高所造成的脂质过氧化增强有关     

强直性脊柱炎棘上韧带组织病理学和超微结构观察

目的 观察强直性脊柱炎(AS)棘上韧带在脊柱骨性强直后发生的组织病理学及超微结构改变.方法 通过光镜下观察苏木素-伊红(HE)、苦味酸-天狼星红染色,并行电镜观察微观结构改变,与对照组进行比较.图像处理软件分析血管密度、胶原原纤维分布密度.采用t检验进行统计学分析.结果 实验组光镜下,胶原纤维排列紊乱,可见玻璃样变及脂肪浸润,大量血管增生,管径不一,有Ⅲ型胶原表达,电镜下胶原纤维分布稀疏,周期性横纹模糊,基质降解,可见基质小泡.对照组胶原纤维排列规则,无上述改变.实验组血管密度(6.8±0.7)/HP,对照组(1.0±0.4)/HP;实验组胶原原纤维分布密度(122±22)/μm2,对照组(218±10)/μm2,2组差异有统计学意义(P＜0.05).结论 棘上韧带在AS疾病进展过程中出现一系列的组织病理学改变.     

Electron microscopic analysis of sequential liver biopsy samples from patients with rheumatoid arthritis. Correlation with light microscopic findings

We used electron microscopy (EM) to analyze 52 biopsy samples from 22 patients who were receiving long-term weekly oral doses of methotrexate (MTX) for the treatment of rheumatoid arthritis. Forty-eight biopsy samples were obtained after 2–6 years of continuous treatment, and 4 samples were obtained before treatment was begun. Specimens were graded for neutral fat, secondary and tertiary lysosomes, and smooth endoplasmic reticulum (SER) in hepatocytes, and for collagen in the perisinusoidal space (Disse's space). We examined the correlations between the EM findings and the light microscopic (LM) findings in the same biopsy specimens, and between the EM findings and the results of simultaneous monthly measures of aspartate transaminase, alkaline phosphatase, bilirubin, and albumin levels, as well as history of alcohol consumption before MTX treatment and monthly assessments of clinical status during the course of treatment. The presence of collagen was minimally increased in these sequential biopsy samples, whereas fat, lysosomes, and SER were decreased. The SER decrease was statistically significant. EM findings of collagen in the space of Disse did not correlate with early fibrotic changes observed with LM. Thus, after as long as 6 years of weekly oral treatment with MTX, hepatic ultrastructural changes are minimal and are not clinically significant. The use of EM for sequential biopsy studies allows the quantitation of long-term hepatic changes that may be more limited than the impression gained after LM analysis.     

Calcium pyrophosphate dihydrate crystal deposition in synovium. relationship to collagen fibers and chondrometaplasia

Objective. Reasons for apparent primary deposition of calcium pyrophosphate dihydrate (CPPD) crystals in some synovial membranes have not been systematically examined. We undertook the present study to investigate for and compare possible cellular and matrix factors related to the presence of these crystals in synovium and cartilage. Methods. Ten synovial membrane specimens and 6 cartilage specimens were obtained at the time of joint surgery from 10 patients with CPPD crystal deposition disease, for light microscopic (LM) and electron microscopic (EM) studies. Results. In all synovial and cartilage specimens, we found many of the small CPPD crystals aligned on or in parallel to collagen fibers, as seen by EM. In 9 of the 10 crystal-containing synovia, we found foci of chondrometaplasia adjacent to CPPD, by LM. In 7 of the synovia, including the one without LM evidence of chondrometaplasia, we observed the presence of chondrocyte-like cells by EM. We did not note any predictable relationship between the crystals and matrix vesicles, either in synovium or in cartilage. Conclusion. Our EM findings provide evidence of the relationship of small CPPD-like crystals, presumably early forms, to collagen fibers both in synovium and in cartilage. By LM and EM, we also demonstrate evidence of a close association between chondrometaplasia and CPPD deposits in synovium. We suggest that chondrometaplasia might be responsible for synovial CPPD formation in predisposed patients. Both the collagen fibers and chondrocyte-like cells seem to be involved in the primary formation of CPPD deposits in the synovium as well as in the cartilage.     

Atrial electrical and structural abnormalities in an ovine model of chronic blood pressure elevation after prenatal corticosteroid exposure: implications for development of atrial fibrillation.

AIMS: Elevated blood pressure (EBP) is the most prevalent and potentially modifiable risk factor for AF, yet little is known of its atrial effects. We aimed to characterize the atrial electrical and structural changes in a chronic ovine model of EBP after prenatal corticosteroid exposure. METHODS AND RESULTS: Twelve sheep with chronically EBP (mean arterial pressure 94+/-3 mmHg) and six controls (71+/-4 mmHg, P<0.01) underwent acute open chest electrophysiologic and pathologic studies. We measured refractoriness at the atrial appendages at 3 cycle lengths (CL); conduction velocities at Bachmann's bundle, both atrial appendages and free walls at 4 CLs; conduction heterogeneity; atrial wavelength and AF duration. We performed light microscopy (LM) and electron microscopy (EM) and collagen and apoptosis studies. EBP was associated with widespread conduction abnormalities, shortening of atrial wavelength, and increased AF. There was no significant change in refractoriness. LM demonstrated atrial myocyte hypertrophy and myolysis in all EBP sheep and focal scarring in six. EM demonstrated mitochondrial and nuclear enlargement and increased collagen fibrils in EBP sheep, findings not present in any controls. Atrial collagen and apoptosis were increased in EBP animals. CONCLUSION: This study demonstrates that chronically, EBP is associated with significant atrial electrical and structural remodelling. These changes may explain the increased propensity to atrial arrhythmias observed with long-standing EBP.     

Phosphorylcholine-coated stents in porcine coronary arteries: in vivo assessment of biocompatibility

AIMS: To examine the angiographic (quantitative coronary angiography), morphometric, light microscopic (LM) (i.e., histology and immunohistochemical staining) and electron microscopic (EM) findings after implantation of phosphorylcholine (PC)-coated compared to uncoated stents in porcine coronary arteries. METHODS: Forty (25 PC-coated, 15 uncoated) divYsio stents were implanted into the coronary arteries of 20 pigs. Quantitative coronary angiography (QCA) was performed pre-stent and post-implantation in fifteen pigs, at 28 days. Two pigs were killed at 5 days (LM and scanning EM), one pig at 14 days (scanning EM) and 17 pigs at 28 days (LM, scanning EM, transmission EM). At 28 days, thirty-two of 34 stented segments excised were formalin-fixed, of which 30 were embedded in resin and sectioned for morphometry and LM. Remaining stents were examined by TEM and SEM. RESULTS: No angiographically occlusive thrombosis occurred in any of the stents. LM at 5 days showed endothelialization of PC-coated and uncoated stents, which was also confirmed by scanning EM at 14 days. At 28 days, QCA and morphometry showed no significant differences between PC-coated and uncoated stents. A few inflammatory cells were seen in both stent types at 5 days but there was no inflammatory or additional tissue reaction to PC-coated compared to uncoated stents at 28 days. CONCLUSIONS: The divYsio stents, with or without PC coating, performed equally well in terms of acute patency, 28-day QCA and morphometry. The PC coating allows a stent to endothelialize normally and is not associated with specific histological changes. The PC coating on the divYsio stent appears biocompatible.     

Light and electron microscopic analysis of liver biopsy samples from rheumatoid arthritis patients receiving long-term methotrexate therapy

OBJECTIVE: We study liver damage in forty-two patients with rheumatoid arthritis (RA) using light (LM) and electron microscopy (EM) and assess histological changes after four years of treatment with methotrexate (MTX). PATIENTS AND METHODS: liver biopsies (LB) were taken before and after four years of treatment. Patients received weekly doses of between 7.5-15 mg of MTX. RESULTS: Fourteen per cent of the baseline LB presented mild perisinusoidal fibrosis (Roenigk IIIA) and the rest a lower Roenigk grade; EM identified an increase in collagen fibers in the Disse spaces in 50% of baseline LB. Neither microscopy technique revealed histological progression in any of the sequential LB. Variables that correlated with histological abnormalities were patient's age, length of evolution of the disease, alcohol consumption and biochemical data (gammaglutamate transferase and albumin); the cumulative dose of MTX was not correlated with worse histological findings. Correlation between the two microscopy techniques was good, though EM was more sensitive than LM for the detection of fibrosis. CONCLUSIONS: RA patients present with liver damage before treatment with MTX. The alterations are mild. At low doses MTX treatment is safe. In addition to the recommendations of the American College of Rheumatology, other factors associated with liver impairment are patient's age and length of evolution of the RA.     

丹参注射液及血栓通注射液对缺血再灌注大鼠肾脏的影响

本文研究了丹参注射腋（ＳＭ）及以三七皂甙为主要成分的血栓通注射液（ＡＲ）对缺血再灌注大鼠肾脏的影响。将Ｗｉｓｔａｒ大鼠肾蒂夹闭造成缺血１ｈ再灌注前１０ｍｉｎ分别给大鼠尾静脉注射ＳＭ及ＡＲ，于再灌注１５ｍｉｎ测定肾皮质ＳＯＤ，ＧＳＨｐｘ活力及ＧＳＨ，ＧＳＳＧ含量。于再灌注２４ｈ后检查大鼠血肌酐含量，并进行病理学检查。结果发现，ＳＭ和ＡＲ起到了相当于活性氧清除剂的作用，显著地改善了大鼠肾脏的再灌注性损伤，无论是肾脏的功能或形态均较未用药组有显著的改善；肾皮质ＳＯＤ及ＧＳＨｐｘ的活力亦受到有效的保护；肾皮质ＧＳＨ含量显著低于未用药组。     

Cell adhesion and migration of different human colon cell lines and primary tumors

Summary Several cell lines derived from colon tumors (HT-29, WiDr, PT4, PT5) and from human liver metastases (LM3) and primary human colon tumor cells (PTR) were compared with regard to their ability to migrate and to attach to different substrates (collagen G, laminin, fibronectin). Cells from the WiDr cell lines migrated actively, whereas the other cell lines, and LM3 and the PTs showed almost no migratory activity. The attachment efficiency was best in all cell lines assayed when tested on collagen followed by laminin and fibronectin as substrates. The HT-29 cells showed the strongest adhesion to all substrates, while the adhesiveness of PT4, PT5, and LM3 was reduced. The WiDr cells which migrated best showed the lowest adhesion to the substrates.     

LIGHT, IMMUNOFLUORESCENCE AND ELECTRON MICROSCOPY RENAL BIOPSY FINDINGS AS PREDICTORS OF MORTALITY IN EIGHTY-FIVE SPANISH PATIENTS WITH SYSTEMIC LUPUS ERYTHEMATOSUS

This study aimed to define prognostic indicators of death inrenal biopsies from Spanish patients with SLE. Renal biopsiesof eighty-five lupus patients with and without clinical nephritis,taken betweeen 1974 and 1987, were reviewed. Samples previouslyprocessed for light (LM), immunofluorescence (IM) and electron(EM) microscopy were analysed blind. Kaplan-Maier curves, log-ranktest, and multivariate Cox's regression statistical methodswere used for comparison of biopsy data in relation to patientsurvival. Univariate analysis showed that vascular hyalinosis,glomerular sclerosis, fibrous crescent and chronicity indexhigher than 3 by LM, and intramembranous dense-deposits by EMare predictors of poor survival. A multivariate approach confirmedthe independent influence of vascular hyalinosis, chronicityindex higher than 3 and intramembranous deposits. A predictivemodel can be constructed with three LM (hyalinosis, tubularatrophy and glomerular sclerosis) and three EM variables (subepithelial,mesangial and intramembranous deposits). Selected renal biopsychanges detected by LM and EM are therefore predictors of deathin patients with lupus. Chronicity markers, more than thoseof activity or severity, are the best prognostic indicators. KEY WORDS: Lupus nephritis, Predictors of mortality in SLE, Survival in lupus nephritis, Electron microscopy, Light microscopy     

Morphometric estimation of regional differences in the rat lung

To provide a safe basis for the sampling of tissue in future morphometric investigations of the rat lung, we searched for quantitative regional differences in pulmonary structure at light microscopic (LM) and electron microscopic (EM) levels. The lungs of 11 male rats about 6 weeks of age were fixed by standard intratracheal instillation of glutaraldehyde in the supine position and embedded either in paraffin for LM or in epoxy resin for EM investigation. Sampling of tissue was designed to test for differences between lobes and between central and peripheral lung parenchyma. LM morphometry was performed by manual point counting and by using a version of an improved automated image analyzer, Quantimet 720. EM morphometric results were obtained by manual point and intersection counting only. LM point counting showed that the proportion of parenchyma was highly constant in all lobes, varying only between 79.9% and 81.5%. In the left lung, which was partitioned into two equal halves, the amount of parenchyma was significantly lower in the apical region (mean values, 72.6% compared to 83.1%; p less than 0.002), which regularly contained the hilum. Quantimet analysis of central and subpleural lung portions revealed intralobar differences. The volume density of interalveolar septa and the air space surface density were significantly decreased in subpleural compared to central lung regions (by 7% and 4.6%, respectively). EM morphometry demonstrated that the interalveolar septa were evenly structured in all lobes except for the harmonic mean thickness of the air-blood barrier, which was lower in upper lobes. In addition, the volume density of interstitial cells was found to be significantly increased in central compared to peripheral parenchyma. The results indicate that for quantitative LM analysis the smallest possible sampling unit is an entire lobe. For EM morphometry, the often practiced approach to consider information drawn from one lobe representative for the whole lung seems to be appropriate for most parameters. In view of the structural differences between central and peripheral lung parenchyma, however, attention has to be paid to applying a properly weighted sampling procedure. Depending on the size of the lobe, the peripheral mantle (2 mm thick) can represent up to 75% of the lobar volume.     

Clinical significance of corticosteroid therapy for eosinophilic myocarditis

The recommended treatment for eosinophilic myocarditis (EM), pathologically defined as myocardial inflammation with eosinophil infiltration, is corticosteroids. Although EM has a wide variety of clinical features including the degree of eosinophilic infiltration, there have been no reports on how patients with EM should be treated with corticosteroids irrespective of their pathological findings.Thirty-seven consecutive patients with acute myocarditis hospitalized in our institute between 1996-2009 were enrolled. Excluding those with secondary EM such as Loeffler's endocarditis, hypereosinophilic syndrome, and Churg-Strauss Syndrome, together with drug-induced allergic myocarditis, the subjects were divided into 2 groups according to the existence of eosinophils in the myocardial interstitium observed in endomyocardial biopsy specimens. There were no differences in the clinical characteristics on admission between the 2 groups: with (group EM, n = 22) and without (group lymphocytic myocarditis (LM), n = 7) eosinophilic infiltrates irrespective of pathological differences. The treatment policy has been consistent in our institution: intensive hemodynamic observation and support without corticosteroid administration, not only in LM but also in idiopathic EM. There was no significant difference in clinical recovery in the acute phase as indicated by the hospitalization period, left ventricular ejection fraction, or long-term prognosis in EM compared to LM.A conventional management strategy for idiopathic EM without corticosteroid administration can improve the prognosis in the acute and chronic phases, similar to that of LM.