Ventral specification and perturbed boundary formation in the mouse midbrain in the absence of Hedgehog signaling.

Although Hedgehog (HH) signaling plays a critical role in patterning the ventral midbrain, its role in early midbrain specification is not known. We examined the midbrains of sonic hedgehog (Shh) and smoothened (Smo) mutant mice where HH signaling is respectively attenuated and eliminated. We show that some ventral (Evx1+) cell fates are specified in the Shh-/- mouse in a Ptc1- and Gli1-independent manner. HH-independent ventral midbrain induction was further confirmed by the presence of a Pax7-negative ventral midbrain territory in both Shh-/- and Smo-/- mice at and before embryonic day (E) 8.5. Midbrain signaling centers are severely disrupted in the Shh-/- mutant. Interestingly, dorsal markers are up-regulated (Wnt1, Gdf7, Pax7), down-regulated (Lfng), or otherwise altered (Zic1) in the Shh-/- midbrain. Together with the increased cell death seen specifically in Shh-/- dorsal midbrains (E8.5-E9), our results suggest specific regulation of dorsal patterning by SHH, rather than a simple deregulation due to its absence.     

Arthrobacter scleromae sp. nov. isolated from human clinical specimens

A gram-positive, coryneform bacterium was isolated from swollen scleromata of a dermatosis patient. An analysis of its phenotypic, chemotaxonomic, and genotypic characteristics showed that this bacterium is closely associated with Arthrobacter oxydans and Arthrobacter polychromogenes but that it belongs to a distinct species, for which the name Arthrobacter scleromae sp. nov. is proposed.     

Ultrastructure and chromogranin A immunogold labelling of ECL cell carcinoids

Poorly differentiated neuroendocrine cells can be difficult to recognise. Sensitive methods are needed to label cells that have lost their ultrastructural features and have reduced concentrations of neuroendocrine markers. In gastric neoplasms, enterochromaffin-like cells might dedifferentiate and lose their characteristic granules and secretory vesicles, making detection of such cells increasingly difficult. However, chromogranin A (CgA) immunogold labelling could provide sensitive and specific detection of gastric neuroendocrine cells. We present ultrastructural findings, CgA immunogold labelling as well as conventional immunohistochemical findings of two human enterochromaffin-like cell carcinoids. Electron-dense granules of poorly differentiated cells were less intensely labelled than granules in well-differentiated cells. Granules with atypical shape as well as punctuate granules previously found in neuroendocrine neoplasms were also CgA labelled. The CgA labelling efficacy after antigen retrieval in an alkaline solution was higher after heating in an autoclave at 135 degrees C compared to a microwave at 100 degrees C for both granules and secretory vesicles without significant deterioration of the ultrastructure. In conclusion, the use of CgA immunogold labelling could ensure a specific classification of cells with neuroendocrine granules and be a supplement to immunohistochemical examination of poorly differentiated tumours.     

Ultrastructural study of a muscle biopsy in a case of GM1 gangliosidosis type I.

The main ultrastructural findings in a muscle biopsy from a child aged 11 months with a GM1 gangliosidosis were cytoplasmic inclusions of two different types: (1) inclusions filled with a moderate electron dense and polymorphous material thought to correspond to ganglioside accumulation and lying only in the Schwann cells of intramuscular nerves. (2) Vacuolar inclusions regarded as containing polysaccharides and observed in perineurial cells, endothelium and pericytes of blood vessels, and also in muscle satellite cells. The muscle fibres only exhibited moderate and non-specific changes. The study shows that in a muscle biopsy of GM1 gangliosidosis the two characteristic types of storage deposits and their preferential localization in different cells may be demonstrated, providing that the intramuscular nerves and motor end plates are examined.     

磺化聚醚砜对光敏剂-亚甲蓝的吸附去除研究Ⅱ.磺化聚醚砜对血浆中亚甲蓝的吸附去除研究

研究了磺化聚醚砜吸附柱对血浆中亚甲蓝的吸附效果,并检测了流经吸附柱的牛血清白蛋白随时间变化的规律以及过柱前后血浆中主要生化指标的变化情况.结果:磺化聚醚砜对亚甲蓝的吸附效果明显优于聚醚砜对亚甲蓝的吸附效果;其对白蛋白的吸附较小;对血浆中主要生化指标的影响可以忽略不计.     

Activation of Classical Complement Pathway by Naturally Occurring Heteroantibodies in Normal Rabbit Serum. Effect on Subpopulations of Human Lymphoid Cells

Heteroantibodies present in normal rabbit serum (NRS) are toxic to human B lymphocytes, T lymphocytes, and monocytes. Even NRS, which exhibits little back ground cytotoxicity for human lymphoid cells in conventional HLA or B-cell lymphocytotoxic assays, can be shown to contain considerable activity by making two modifications in usual procedures: by washing cells in saline or balanced salt solutions devoid of protein or sugar substances, and by increasing incubation time for 1 h to 3--4 h. Using such modifications, the cytotoxic activity of NRS towards human lymphoid cells was investigated and was found to involve activation of the classical complement pathway rather than activation of the alternate complement pathway. Residual unwanted background cytotoxicity of NRS toward human lymphoid cells can be decreased without loss of desired complement activity either by heating NRS for 15 min at 50 degrees C or by mixing NRS with small amounts of normal human serum.     

Role of excitatory amino acid transporter 1 in neonatal rat neuronal damage induced by hypoxia-ischemia

The role of excitatory amino acid transporter 1 in neonatal rat neuronal damage was studied following hypoxia-ischemia. To induce hypoxia-ischemia injury, rats on postnatal day 7 were exposed to 8 % oxygen for 2 h following unilateral common carotid artery ligation. According to brain damage scoring based on Cresyl Violet staining, the neuronal damage time-dependently changed in the ischemic regions following hypoxia-ischemia. Immunohistochemical studies showed that excitatory amino acid transporter 1 expression was mainly observed in the cerebral cortex ipsilateral to common carotid artery ligation and markedly increased at 24 h and 48 h following hypoxia-ischemia. Combined with confocal laser scanning microscopic analysis, double staining showed that excitatory amino acid transporter 1 positive staining appeared in neurons as well as astrocytes after hypoxia-ischemia. Most excitatory amino acid transporter 1 positive staining cells exhibited regular morphological characteristics and only a few were double-stained by terminal deoxynucleotidyl transferase-mediated deoxyuridinetriphosphate nick-end labeling. Down-regulation of excitatory amino acid transporter 1 expression by intraventricular administration of specific antisense oligonucleotide exacerbated neuronal damage in hypoxia-ischemia brain. These results suggest that the increase of excitatory amino acid transporter 1 expression may be involved in a pathophysiological process of hypoxia-ischemia brain damage and may reflect a self-compensative mechanism for protecting neurons from further injury.     

Comparison of assays to detect cytomegalovirus shedding in the semen of HIV-infected men

We sought to determine the optimal assays for cytomegalovirus (CMV) shedding in semen. Over a 2-month period, 149 HIV-1-infected men who have sex with men each provided up to three semen specimens. Specimens were tested for CMV by culture, rapid assay (shell vial) and polymerase chain reaction (PCR). By culture, 30% of seminal plasma and 28% of seminal cell specimens grew CMV. By rapid assay, results were 38 and 33%, respectively. By PCR, 56% of seminal cell specimens demonstrated CMV: 20% in a single semen specimen; 33% in two specimens; and 34% in all three specimens. Overall, 69% of men had CMV detected by PCR in at least one seminal cell specimen. By quantitative PCR, 14% had ten, 14% had 100, 16% had 1000, and 12% had 10 000 copies in 6.25 μl of semen analyzed. Adjusting for initial CD4+ cell count, men with CMV shedding demonstrated by PCR at the first visit were approximately four times as likely to shed CMV at a subsequent visit (RR 4.28, CI: 2.30–7.95). CMV shedding was associated with decreased CD4+ cell counts in peripheral blood (P=0.05). It is concluded that the PCR assay provided the greatest sensitivity among the three detection methods.     

一步温育EIA法检测HBsAg

本文报道用尼龙网为载体的抗ＨＢｓＡｇ抗体膜作免疫吸附剂用于酶联免疫测定，对检测ＨＢｓＡｇ的免疫反应程序和反应如酶标／抗原溶液配比量和温育反时间等进行了研究。结果得到一步温充ＥＩＡ法的灵敏度与二步温育ＥＩＡ法相比没有下降，检测时间由原来的约３ｈ缩短至约５０ｍｉｎ。     

低频超声透皮给药的研究进展与应用

低频超声可以增强包括大分子药物在内的许多药物的透皮传输,其主要机制是超声的空化作用,大多数人认为是通过改变角质层角化细胞排列结构来提高皮肤渗透能力的.低频超声透皮给药已被人们用于离体实验和动物活体实验,到目前为止,无论是小分子透皮传输还是大分子透皮传输都有很多成功的例子.但是真正通过低频超声透皮导入药物进行治疗的临床应用报道很少,需要更进一步大量的临床试验以确定其安全性与实用价值.一旦其安全性得以证实,合适的低频超声透皮仪研制成功,低频超声快速透皮必将成为一种安全、有效、可控、经济的新型给药方式.