Age-dependent DNA methylation changes in the ITGAL (CD11a) promoter

DNA methylation patterns change with age in a complex fashion, typically with an overall decrease in genomic deoxymethylcytosine (d(m)C) content, but with local increases in some promoters that contain GC-rich sequences known as CpG islands. While the consequences of age-dependent CpG island methylation have recently been studied in organs such as the colon, less is known about the functional significance of the progressive hypomethylation of promoters lacking CpG islands, and the significance of age-dependent changes in T cell DNA methylation is completely unexplored. We asked if age-dependent DNA hypomethylation might contribute to overexpression of the T cell ITGAL gene, which encodes CD11a, a subunit of LFA-1. CD11a mRNA increased with age as well as with experimentally induced DNA hypomethylation. This increase correlated with hypomethylation of sequences flanking the ITGAL promoter in vitro and in aging. 'Patch' methylation of the region suppressed promoter function. DNA methyltransferases 1 and 3a also decreased with aging. These results indicate that hypomethylation of regions flanking the ITGAL promoter may increase CD11a expression, and suggest that age-dependent hypomethylation of promoters lacking CpG islands, perhaps due to decreased DNA methyltransferase expression, may be one mechanism contributing to increased T cell gene expression with aging.     

远端蒂腓肠神经营养血管皮瓣与肌皮瓣的临床应用与改进

目的:报道应用远端蒂腓肠神经营养血管皮瓣,肌皮瓣修复小腿下段及足踝部软组织缺损的可行性安全性和临床效果。方法:对42例以远端蒂腓肠神经营养血管(肌)皮瓣修复小腿下段及足踝部不同原因所致软组织缺损病例进行总结分析。本组男36例,女6例;年龄最大75岁、最小6岁;皮瓣最大面积17.0cm×15.0cm,最小6.0cm×5.0cm,其中12例皮瓣面积在10.0cm×10.0cm以上;6例设计为肌皮瓣(腓肠肌外侧头),肌瓣最大为10.0cm×7.0cm×2.0cm,最小为6.0cm×5.0cm×1.0cm。结果:所有病例术后皆出现不同程度的皮瓣肿胀,暗道较明道者明显。2例大皮瓣经行小隐静脉远端结扎仍出现肿胀、色暗,皮瓣近侧1/3坏死。皮瓣边缘坏死3例,换药治愈。部分坏死需行植皮者3例。36例术后伤口I期愈合,骨外露软件组织缺损覆盖修复满意,6例II期愈合,其中糖尿病,地中海贫血各一例。结论:(1)远端蒂腓肠神经营养血管皮瓣转位修复小腿下1/3及足踝部缺损创面,极有临床实用价值;(2)设计切取腓肠神经营养血管肌皮瓣修复小腿及足踝填充感染创腔是可行的;(3)但对其皮瓣及所携带的肌瓣究竟切取多大面积是安全的、肌瓣的血运机理以及远端蒂筋膜皮瓣中小隐静脉干是否结扎,何处结扎等问题仍有待进一步研究。     

A rat model for radiation-induced proctitis

Radiation proctitis is a frequent acute complication encountered with pelvic irradiation. This study was aimed at establishing the optimal radiation dose for radiation-induced proctitis in rats. Female Wistar rats were used. The rectal specimens were examined morphologically at 5th and 10th day following 10-30 Gy irradiation in single fraction. With increasing dose, mucosal damage became worse, and there was a prominent reaction after > or =15 Gy. We selected 17.5 Gy as an optimal dose for radiation proctitis and examined specimens at day 1-14 and at week 4, 6, 8, and 12 after 17.5 Gy. The rectal mucosa revealed characteristic histological changes with time. An edema in lamina propria started as early as 1-2 days after irradiation and progressed into acute inflammation. On day 7 and 8, regeneration was observed with or without ulcer. Four weeks later, all regeneration processes have been completed with end result of either fibrosis or normal appearing mucosa. This study showed that the radiation injury of the rectum in rat develops in dose-dependent manner as it has reported in previous studies and suggested that 17.5 Gy in single fraction is the optimum dose to evaluate the protective effect of various medications for radiation proctitis in face of the clinical situation.     

慢性乙肝患者杀伤性免疫细胞功能的研究

通过对４４例病毒性肝炎患者Ｔ细胞亚群，ＮＫ细胞活性与ＬＡＫ细胞活性的观察，探讨了在慢性乙型肝炎病毒复制与非复制状态下的杀伤性细胞活性。结果表明：在乙肝病毒的高复制状态下，ＣＤ８＾＋细胞数增加，ＣＤ４＾＋／ＣＤ８＾＋比例显著下降；ＮＫ细胞活性与ＬＡＫ细胞活性也明显低下，且在ＨＢｅＡｇ与ＨＢＶＤＮＡ阳性组中，ＮＫ活性与ＬＡＫ活性的改变与ＨＢｅＡｇ的Ｐ／Ｎ值变化呈显著负相关，而ＮＫ活性与ＬＡＫ活性变化则     

Negative pressure effects during the isothermal crystallization of isotactic poly(propylene) studied by light and atomic force microscopy

The formation of holes during the late stage of the isothermal crystallization in thin films of isotactic poly(propylene) between two cover glasses was observed by light microscopy and atomic force microscopy. This behavior can be described consistently by the well-known negative pressure effect. Light microscopy reveals the simultaneous and sudden occurrence of a large number of small holes at the liquid-solid interface after the liquid in front of the spherulites is completely confined by other spherulites for a certain time interval. In exceptional cases only a few holes appear and finally large cavities are formed. Atomic force microscopy measurements carried out in the height mode are able to prove the hole formation in front of the spherulites. Furthermore, a substantial thinning of the two-dimensional spherulites in thin films can be observed prior to the hole formation.     

Emphysematous lung destruction by cigarette smoke. The effects of latent adenoviral infection on the lung inflammatory response.

This study was designed to test the hypothesis that cigarette smoke-induced inflammation and emphysema are amplified by the presence of latent adenoviral (Ad) infection, and to determine whether this emphysematous process can be reversed by all-trans-retinoic acid (RA) treatment. The results confirm that in guinea pigs, chronic cigarette-smoke exposure caused lesions similar to human centrilobular emphysema. They also show that latent Ad infection combined with cigarette-smoke exposure caused an excess increase in lung volume (P < 0.001), air-space volume (P < 0.001), and lung weight (P < 0.01), and further decrease in surface-to-volume ratio (P < 0.001) compared with smoke exposure alone. RA treatment failed to reverse these emphysematous changes. Analysis of inflammatory response in parenchymal and airway tissue showed that smoking caused an increase of polymorphonuclear leukocytes (PMNs) (P < 0.0002), macrophages (P < 0.001), and CD4 cells (P < 0.0009), and that latent Ad infection independently increased PMNs (P < 0.001), macrophages (P = 0.003), and CD8 cells (P < 0.001). We conclude that latent Ad infection amplifies the emphysematous lung destruction and increases the inflammatory response produced by cigarette-smoke exposure. In this study, the increase in CD4 was associated with cigarette smoke and the increase in CD8 cells with latent Ad infection.     

Isometric and isokinetic torque curves at the knee joint

Isometric and isokinetic torques of bilateral quadriceps and hamstrings were measured with Isokinetic Rehabilitation and Testing System (Model No. Cybex 340) on 40 normal untrained subjects, 20 males and 20 females, ranging between the ages of 23 and 35 years. The mean peak isometric and isokinetic torque values of both muscle groups showed no significant differences between dominant (right) and nondominant (left) limbs in both sexes; however there were significant differences between the male and the female. As the angular velocity increased, the peak torque significantly decreased, and the point of peak torque output occurred significantly later in the range of motion for quadriceps and hamstrings (p less than 0.01). There were no significant changes in the hamstrings to quadriceps (H/Q) ratios as the angular velocity increased. However, there were significant differences of mean H/Q ratio between male and female (p less than 0.01). Height had significant positive correlation with peak isometric and isokinetic torques for both quadriceps and hamstrings (p less than 0.01). Weight was found to correlate significantly with peak isometric and isokinetic torques (p less than 0.01). The mean isometric torques were significantly higher than the mean isokinetic torques for any joint angles in both sexes (p less than .01).     

Significance of cytogenetic and fluorescence in situ hybridization analysis in evaluating antichronic myeloid leukemia efficacy of different immune effector cells

We report on the antileukemia effect of interleukin 2 (IL2) on different immune cells from 22 patients with chronic myeloid leukemia (CML). Bone marrow cells from these patients were first cultured in modified long-term bone marrow culture medium for several days, then separately cultured with lymphokine activated killer cells (LAK), cytokine-induced killer cells (CIK), and dendritic cell cocultured CIK (DC-CIK) for another 1-2 days. They were then detected for presence of the Philadelphia chromosome (Ph) by cytogenetic analysis and fluorescence in situ hybridization (FISH). The percentage of Ph-chromosome-positive cells in the bone marrow mononuclear cells after culturing with CIK and DC-CIK was significantly lower than that after culturing with IL2 or LAK. Our results demonstrate that cytogenetics and FISH are useful techniques for the evaluation of the anti-CML effect of immune cells and that CIK or DC-CIK can be appropriate candidates for adoptive immune cell therapy in vivo or for leukemia cell purging ex vivo.     

Membrane ionic currents and properties of freshly dissociated rat brainstem neurons

It is well known that neuronal firing properties are determined by synaptic inputs and inherent membrane functions such as specific ionic currents. To characterize the ionic currents of brainstem cardio-respiratory neurons, cells from the hypoglossal (XII) nucleus and the dorsal motor nucleus of the vagus (DMX) were freshly dissociated and membrane ionic currents were studied under whole-cell voltage and current clamp. Both of these neurons showed a TTX-sensitive Na⁺ current with a much larger current density in XII than DMX neurons. This Na⁺ current had two (fast and slow) distinct inactivation decay components. The ratio of the magnitudes of the fast to slow component was roughly two-fold greater in DMX than in XII cells. Both DMX and XII neurons also showed a high voltage-activated Ca²⁺ current, but this current density was significantly greater (three-fold) in DMX than XII neurons. A relatively small amount of low-voltage activated Ca²⁺ current was also observed in DMX neurons, but not in the majority of XII cells. A transient and a sustained outward current components were observed in DMX cells, but only sustained currents were present in XII neurons. These outward currents had a reversal potential of about − 70 mV with 3 mM external K⁺ and −30 mV with 25 mM K⁺, and substitution of K⁺ with cesium and tetraethylammonium suppressed more than 90% the outward currents, indicating that most outward currents were carried by K⁺. The transient outward current consisted of two components with onesensitive to 4-aminopyridine and the other to intracellular Ca²⁺. In XII neurons, BRL 38227 (lemakalim), an ATP-sensitive K⁺ (K_ATP) channel activator, increased the sustained K⁺ currents by 10% of control, and glibenclamide, a K_ATP channel blocker, decreased the sustained K⁺ currents by 20%. Evidence for the presence of an inward rectifier K⁺ current was also obtained from both XII and DMX neurons. These results on XII and DMX neurons indicate that (1) the methods used to dissociate neurons provide a useful means to overcome voltage clamp technical difficulties; (2) ion channel characteristics such as density and biophysical properties of DMX neurons are very different from those of XII neurons; and (3) several newly discovered membrane ionic currents are present in these cells.