Histological and biological developmental characterization of the human cerebellar cortex

The aim of this study was to investigate the histological and biological features of the human cerebellar cortex development and differentiation. We analyzed 52 brains of fetal and infant death victims, aged from 17 gestational weeks to 12th postnatal month. In particular, in the cerebellar cortex at different ages we evaluated, besides the structural aspects, the expression of several biomarkers implicated in proliferative processes (c-fos, PCNA and apoptosis). We observed morphological patterns progressively evolving every month, from the indefinite structure of the second gestational trimester to the four-layered structure (external granular layer, molecular layer, Purkinje cell layer, internal granular layer) of the late fetal cortex and subsequently to the three-layered postnatal definitive morphology, due to involution of the external granular layer. The evaluation of the biological features of the cerebellar cortex showed high proliferative activity mainly confined to the transient external granular layer in prenatal life, and high apoptotic index after birth. Thus, the histological examination, better with the support of biomarker investigations, allows with accuracy to describe the dynamic sequence of steps that occur in human cerebellar cortex development and to establish in each case the age, namely the pre- or postnatal month of life. Consequently, we can diagnose delayed or altered processes of differentiation during the development of the human cerebellar cortex.     

Monolayer cultures of normal human bone cells contain multiple subpopulations of alkaline phosphatase positive cells

Summary Cytochemical staining of normal human bone cells in monolayer cultures for alkaline phosphatase (ALP) indicated that the cultures contained mixed-cell populations. Time course evaluations of the cytochemical staining revealed, in addition to the ALP-negative cell population, at least two subpopulations of ALP-positive human bone cells with different levels of ALP. A cytochemical method has been developed which separates the ALP-positive cells into high and intermediate ALP subpopulations. In this method, human bone cells were stained for ALP using an azo-dye method and incubating at 4°C for 10 and 30 minutes, respectively. We defined the cell population that stained positively for ALP at 10 minutes as strong ALP-positive cells, and both strong and intermediate cells were stained at 30 minutes. The intermediate cells were determined from the difference between the values at the two time points. The intra- and interassay variations of the assay, with the same investigator in blinded investigations, were both less than 10% and the interobserver variation was approximately 25%. Analysis of the distribution of ALP levels in cells with a laser densitometer confirmed the presence of at least three cell subpopulations. 1,25(OH)₂D₃ treatment increased the proportions of both ALP-positive cell populations, whereas TGF-beta treatment increased only the intermediate ALP-positive cell population. On the contrary, fluoride increased the proportion of the strong ALP cells, and IGF-1 had no effect on the proportions of either ALP-positive subpopulation. When the ALP-specific activity was compared with the percentage of each ALP-positive subpopulations for the cells treated with effectors, the ALP-specific activity correlated with the total ALP-positive and with the strong ALP-positive populations but not with the intermediate ALP-positive subpopulation. In summary, this study represents the first evidence that normal human bone cells in monolayer cultures contained at least two subpopulations of ALP-positive cells, and that bone cell effectors could have differential effects on each cell population.     

Active production of anti-human T-lymphotropic virus type I (HTLV-I) IgM antibody in HTLV-I-associated myelopathy

We investigated the presence of anti-human T-lymphotropic virus type I (HTLV-I) IgM in sera and cerebrospinal fluid from patients with HTLV-I-associated myelopathy (HAM) by Western blot analysis. Analyses of 36 serum samples revealed that most patients (31/36; 86.1%) had anti-HTLV-I IgM, whereas only four of 23 (17.4%) HTLV-I carriers had it. In studies of cerebrospinal fluid, anti-HTLV-I IgM was detected in 24 of 36 (66.7%) HAM patients, whereas none was detected in nine HTLV-I carriers. The differences were statistically significant (p less than 0.01). These results suggest that persistent active replication of HTLV-I occurs in the central nervous system as well as in the peripheral blood of HAM patients, and may contribute to the development of HAM.     

Multivariate comparison of elemental concentrations in human teeth

R-mode factor analysis was applied to characterize the chemical composition of human teeth investigated by particle induced X-ray emission (PIXE), Rutherford backscattering spectrometry (RBS) and X-ray fluorescence (XRF) techniques. The approach developed in this study enabled the separation between essential mineral teeth components and the pollutants deposited in teeth tissues during the human life. The three independent sources of metals incorporated in human teeth were found. The first source, representing about 43% of the variance of the concentration data, was characterized by pollutant elements of power industry emissions. The second factor was loaded with toxic elements of general urban pollution. The third factor represented the tooth source as it contained mainly large fractions of the mineral components of the tooth tissue as Ca and K.     

酶联免疫吸附试验、间接荧光抗体试验、间接免疫过氧化物酶染色试验诊断人体旋毛虫病的初步研究

用ELISA、IFA和IIP试验检测11例旋毛虫病人血清特异性抗体,阳性率分别为72.72％、81.82％和81.82%。它们之间无统计学差异,3项试验结果间存在良好一致性。 同时检测了30份健康献血员血清和20例其他寄生虫病人血清(包括华支睾吸虫病、四川肺吸虫病、日本血吸虫病、包虫病和阿米巴肝脓肿),旋毛虫病人组的ELISA、IFA和IIP阳性率明显为高。 由于3项免疫学试验均具有较好的特异性和灵敏性,故可以单独或联合用于人体旋毛虫病的诊断和流行病学调查。     

一种较好的RNA分离制备方法及其在人胚RNA提取中的应用

以钒基核苷酸复合物为核酸酶抑制剂,从人胚组织中制备总RNA,所得产品几乎不含DNA和蛋白质,经过分离Poly(A)~ mRNA,证明制品有合成cDNA的完整功能,方法比较简捷,适于大量制备以满足分离mRNA 之需,并讨论了此法的优缺点。     

hTERT启动子调控hNIS基因介导肺癌细胞碘摄取和131I治疗的实验研究

目的 构建含人端粒酶反转录酶(hTERT)核心启动子调控的人钠/碘同向转运体(hNIS)基因重组腺病毒,并靶向转染至肺癌A549细胞中特异性表达.探讨hTERT启动子调控的hNIS基因介导放射性碘治疗肿瘤的可能性.方法 应用AdEasy系统构建重组腺病毒Ad-hTERT-hNIS,同时构建巨细胞病毒(CMV)启动子调控的hNIS重组腺病毒Ad-CMV.hNIS作为阳性对照,不含hNIS的重组腺病毒Ad-CMV作为阴性对照.应用反转录.聚合酶链反应(RT-PCR)方法验证hTERT在转染肿瘤细胞中的转录活性,摄碘实验检测表达的hNIS蛋白功能,细胞克隆形成实验评价131I对转染肿瘤细胞的毒性作用.结果 成功构建重组腺病毒Ad-hTERT-hNIS、Ad-CMV-hNIS及Ad-CMV,并经PCR验证正确.RT-PCR证实hNIS cDNA能从Ad-hTERT-hNIS转染的细胞中扩增出来.Ad-hTERT-hNIS和Ad-CMV-hNIS转染的肺癌A549细胞摄碘能力比阴性对照组Ad-CMV转染的细胞分别提高了23和31倍,且摄碘能力可以被NaClO4抑制.Ad-hTERT-hNIS和Ad-CMV-hNIS转染的肺癌A549细胞均可被131I杀死,2组细胞成活率分别为(31.2±1.45)%和(23.6±4.08)%,而阴性对照组和未转染病毒组分别为(89.0 ±2.99)%和(91.2 ±4.63)%.结论 hTERT启动子调控的hNIS重组腺病毒转染肿瘤细胞后,应用131I治疗有望成为一种新的基因靶向治疗手段.     

Human khat (Catha edulis) chewers have elevated plasma leptin and nonesterified fatty acids

In this study, the effect of regular khat (Catha edulis Forsk) chewing (200 and 400 g) in humans on plasma leptin, nonesterified fatty acid, triacyglycerol, and total cholesterol levels was investigated. The results presented show that khat chewing increases plasma leptin concentration particularly in individuals who chew 400 g of khat leaves. The significance of increased plasma leptin is in explaining the underlying mechanism of the observed effects associated with khat chewing such as loss of appetite, decreased body weight, and hyperthermia. The decreased body weight was evident from the significantly lower body mass index of the khat leaves chewers group as compared to the non–khat leaves chewers group (control). Moreover, like leptin, the plasma levels of nonesterified fatty acids were significantly higher in those chewing 400 g of khat leaves. On the other hand, the plasma levels of triacylglycerol were significantly lower in the 2 khat-chewers groups (200 and 400 g of khat leaves), whereas plasma cholesterol levels were not affected by the 2 levels of khat leaves used in this study. The significance of these results may suggest that khat leaves may contain a component(s) that has the ability to reduce body weight via decreasing appetite.     

Mesh fixation with human fibrin glue (Tissucol) in open tension-free inguinal hernia repair: a preliminary report

Background: The Lichtenstein technique for inguinal hernia repair is easy to learn and associated with few complications. However, recent studies have suggested that this technique is inferior to some ‘sutureless’ repair systems in terms of perceived difficulty, operating time, surgeon satisfaction, etc. Methods: We employed a sutureless Lichtenstein technique in 80 consecutive patients with primary unilateral inguinal hernia, to assess patient and trainee surgeon outcomes. Human fibrin glue was used in place of conventional sutures. Results: The mean operating time was 36 min and all patients were discharged 5–6 h after the operation. On a 100-point visual analogue scale, the surgeons rated the difficulty of the operation as low (mean score, 31), and perceived satisfaction as high (mean score, 84). No complications were observed at 12-month follow-up. Conclusion: This study confirms the efficacy of mesh fixation with human fibrin glue, and supports the viability of a sutureless Lichtenstein procedure.