JC virus genotyping using formalin-fixed, paraffin-embedded renal tissues

Recently genotyping of JC virus (JCV) DNA in renal tissue was reported to be useful to identify the geographic origin of unidentified cadavers. In the above study, autopsied tissue samples without storage or stored in a frozen state were used. This study examined JCV DNA sequence modifications caused by formalin-fixation, in an attempt to elucidate whether formalin-fixed, paraffin-embedded tissue samples can also be used to determine the genotypes of JCV DNA in the kidney. In four cases, a 610 bp typing region of the JCV genome was PCR-amplified from renal tissues stored for 1 year in three different states: frozen at -80 degrees C [Amaker, B.H., Chandler, F.W., Huey, L.O., Colwell, R.M., 1997. Molecular detection of JC virus in embalmed, formalin-fixed, paraffin-embedded brain tissue. J. Forensic Sci., 1157-1159], formalin-fixed, paraffin-embedded [Ault, G.S., Stoner, G.L., 1992. Two major types of JC virus defined in progressive multifocal leukoencephalopathy brain by early and late coding region DNA sequences. J. Gen. Virol. 73, 2669-2678], and soaked in 5% formalin [Baksh, F.K., Finkelstein, S.D., Swalskey, P.A., Stoner, G.L., Ryschkewitsch, C.F., Randhawa, P.R., 2001. Molecular genotyping of BK and JC virus in human polyomavirus-associated interstitial nephritis after renal transplantation. Am. J. Kidney Dis. 38 (2), 354-365]. The amplified fragments were cloned, and the resultant clones were sequenced. In frozen samples, single sequences ('original' sequences) were detected in all cases. In formalin-fixed, paraffin-embedded samples, not only the original sequences but also those with 1-6 base substitutions were detected. From formalin-soaked samples, the original sequences and those with 1-5 and 10-13 substitutions were detected. The genotyping of JCV DNA was not hampered by the presence of 1-6 substitutions, but a shift in JCV genotypes was observed in sequences with 10-13 substitutions. Thus, it was concluded that the genotypes of JCV DNA in the kidney can be determined only with specimens stored in a frozen state or formalin-fixed for a short time.     

IL-1β诱导体外培养的大鼠中脑黑质神经元c-jun表达

应用细胞原位杂交技术，观察经重组小鼠白细胞介素-19（IL-1β）处理后的体外培养的新生1d大鼠中脑黑质神经元c－jun基因的表达．结果显示，培养的黑质细胞多为酪氨酸羟化酶阳性神经元，IL-1β可诱导体外培养的黑质神经元c－junmRNA表达，高水平的表达出现在IL-1β处理后2～4h。说明IL－1β有兴奋黑质神经元的作用，并提示黑质神经元上可能存在IL－1β受体．     

胎肝细胞质液对重症病毒性肝炎患者外周血TL－CFU及mIL－2R影响的实验观察

采用半固体一步单层琼脂培养法和单克隆荧光抗体技术分别观察重症肝炎外周血ＴＬ－ＣＦＵ和ｍＩＬ－２Ｒ，发现重症肝炎患者ＴＬ－ＣＦＵ（１０４．４±３２．６）及ｍＩＬ－２Ｒ（３５．６±８．６）较正常人明显降低。在培养体系中加胎肝细胞质液后，无论在重症肝炎组还是在正常组均不能明显地提高ＴＬ－ＣＦＵ，说明胎肝细胞质液不含具生物佐的促ＴＬ－ＣＦＵ因子。但对ｍＩＬ－２Ｒ表达的影响，在重症肝炎组病人，只有在ＰＨＡ存在条件下才能促进ｍＩＬ－２Ｒ的表达，说明胎肝细胞质波含有某种（些）物质能协同ＰＨＡ促进重症肝炎患者外周血淋巴细胞ｍＩＬ－２Ｒ表达。     

黄皮酰胺促钾通道开放

一种新发现的具有促智作用的药物——黄皮酰胺能抑制去甲肾上腺素(NE)或KCl引起的血管平滑肌收缩。本研究旨在应用膜片钳(patch clamp)技术探讨黄皮酰胺对Wistar大鼠尾动脉平滑肌细胞膜钾离子通道的作用。单个平滑肌细胞用酶法分离,以细胞封接方式记录离子通道活动。在细胞池内注入2μM黄皮酰胺后,钾离子通道活动明显增强。用本实验室开发的计算机软件(patch clamp analysis system,Version 1.0)计算分析通道活动的特征参数。     

低强度脉冲电磁场对大鼠骨质疏松的影响

目的：观察低强度脉冲电磁场（pulsed electromagnetic fields，PEMFs）对去卵巢诱导骨质疏松症的大鼠生化指标和骨应力的影响。方法：雌性SD大鼠30只，随机等分为3组（n=10），分别为假手术组（Sham）、骨质疏松模型组（Model）、脉冲电磁场照射组（PEMFs）。经适应4wk后，在25mg·Kg^-1戊巴比妥钠腹腔麻醉下，Model组和PEMFs组摘除双侧卵巢．Sham组找到但不切除卵巢。各实验组均在相同环境下饲养，模型制备4wk后开始治疗，由GZY型低强度低频率脉冲电磁场发生仪产生低频脉冲磁场，根据实验要求，我们使用亥姆霍兹线圈形成均匀磁场，PEMFs组经照射刺激治疗，频率14．3Hz，场强2Gs．日照8h。Model组和Sham组正常饲养。治疗8wk后，对各组大鼠血清、尿液中ALP和Ca以及骨应力进行检测。结果：（1）ALP、Ca检测结果：与Model组相比，PEMFs组ALP值、Ca值差异均有统计学意义（P〈0．05）。其中，血清中ALP值，Model组为（275．16±228，57），PEMFs组为（179．30±87．68）；Ca值，Model组为（2，66±0，13），PEMFs组为（2．52±0．05）。（2）骨应力检测结果：Model组为（923．60±34．15Pa），PEMFs组为（1152．85±118，20Pa），组间差异有统计学意义（P〈0．05）。结论：研究发现，PEMFs对于促进骨重建、提高Ca吸收和骨应力恢复具有积极作用。     

Miscibility and Intermolecular Specific Interactions in Blends of Poly(hydroxyether sulfone) and Poly(N‐vinylpyrrolidone)

Summary: The blends of poly(hydroxyether sulfone) (PHES) with poly(N‐vinylpyrrolidone) (PVPy) were investigated by means of differential scanning calorimetry (DSC) and FTIR spectroscopy. The miscibility of the blend system was established on the basis of the thermal analysis results. DSC showed that the PHES/PVPy blends prepared by casting from N,N‐dimethylformamide (DMF) possessed single, composition‐dependent glass transition temperatures, indicating that the blends are miscible in the entire composition. The experimental glass transition temperatures have higher values than those calculated on the basis of additive behavior; the variation of the glass transition temperatures of the blends was accounted for by the Kwei equation. FTIR studies indicate that competitive hydrogen bonding interactions exist upon addition of PVPy to the system, which were involved in the self‐ and cross‐association, i.e., ? OH···O?S, ? OH···OH of PHES and ? OH···O?C< of PVPy. The FTIR spectra in the range of the sulfonyl stretching vibrations showed that the hydroxyl‐associated sulfonyl groups are partially “set free” upon addition of PVPy to the system. The IR spectroscopic investigation of both the model compounds and the PHES/PVPy blends suggests that the strength of the hydrogen bonding interactions in the blend system increases in the following order: ? OH···O?S, ? OH···OH and ? OH···O?C<.

Plot of glass transition temperature for PHES/PVPy blends as a function of weight fraction of PVPy. The prediction of the Kwei equation yields the values of k = 1 and q = 122.     

Functionally graded calcium phosphate coatings produced by ion beam sputtering/mixing deposition

Ar+ ion beam sputtering/mixing deposition was used to produce thin calcium phosphate coatings on titanium substrate from hydroxyl-poly-calcium sodium phosphate (HPPA) and HPPA + Ti targets. Three types of coatings (one type of monolayer coating and two types of functionally graded coating) were manufactured. It was found that as-sputtered coatings were amorphous. No distinct hydroxyl band was observed in the FTIR spectra, but new absorption bands were determined for CO3(2-), which resulted during the deposition process. Compositional gradients from the surface to the interior (i.e. adjacent to the substrate) were achieved for the functionally graded coatings. Post-deposition heat treatment indicated that the bonding strength between the coating and the substrate was improved by the use of functionally graded structures.     

Vagal-enteric interface: vagal activation-induced expression of c-Fos and p-CREB in neurons of the upper gastrointestinal tract and pancreas

Many gastrointestinal and pancreatic functions are under strong modulatory control by the brain via the vagus nerve. To start identifying location and neurochemical phenotype of the enteric neurons receiving functional vagal efferent input, we activated vagal preganglionic neurons either by electrical or chemical stimulation and examined the expression of phosphorylated CREB (c-AMP response element binding protein) and the immediate early gene c-Fos. There was no spontaneous expression of both markers in the pancreas and considerable spontaneous expression of p-CREB but not Fos in the upper GI-tract. Unilateral electrical vagal stimulation-induced p-CREB was found in 40% of neurons in the head of the pancreas. Fos expression was found in 70-90% of neurons in the esophagus and stomach, in 20-30% of myenteric plexus neurons and 5-15% in submucosal neurons of the proximal duodenum. Double-labeling experiments showed that a majority of pancreatic neurons and about 25-35% of neurons in the stomach and duodenum contain NADPH-diaphorase and that many of these receive functional vagal input. Other neurons that can be vagally activated contain gastrin-releasing peptide or calretinin. Chemical stimulation of the dorsal surface of the caudal brainstem with the stable TRH analog RX77368 resulted in selective activation of vagal efferents with expression of Fos in a small number of gastric myenteric plexus neurons. The results demonstrate the suitability of this method to investigate magnitude and local distribution of vagal input to the enteric nervous system as well as specificity of its neurochemically coded pathways. They represent the first step in the identification of function-specific units of parasympathetic vagal outflow.     

下颌骨的应力迹线与骨小梁构筑的关系

根据下頜光弹的等倾线图,描绘出主应力迹线规迹。在下颌侧位X线照片和去除唇侧及颊侧密质骨板的标本上,观察骨密质和松质的配布,以及骨小梁的排列和方向,试行探讨骨小梁方向与主应力迹线的关系。用光弹法求得的主应力迹线有两个系列:S_1系在下颌体近水平方向,在下颌支近垂直方向分布;S_2系力线与S_1系诸力线呈正交。在标本和X线照片上,主应力迹线规迹不同程度地在骨小梁的排列上有所反映。