CD8+ T cells undergo activation and programmed death-1 repression in the liver of aged Ae2a,b?/? mice favoring autoimmune cholangitis

Primary biliary cirrhosis (PBC) is a chronic cholestatic disease of unknown etiopathogenesis showing progressive autoimmune-mediated cholangitis. In PBC patients, the liver and lymphocytes exhibit diminished expression of AE2/SLC4A2, a Cl⁻/HCO₃⁻ anion exchanger involved in biliary bicarbonate secretion and intracellular pH regulation. Decreased AE2 expression may be pathogenic as Ae2_a,b^−/− mice reproduce hepatobiliary and immunological features resembling PBC. To understand the role of AE2 deficiency for autoimmunity predisposition we focused on the phenotypic changes of T cells that occur over the life-span of Ae2_a,b^−/− mice. At early ages (1-9 months), knockout mice had reduced numbers of intrahepatic T cells, which exhibited increased activation, programmed-cell-death (PD)-1 expression, and apoptosis. Moreover, young knockouts had upregulated PD-1 ligand (PD-L1) on bile-duct cells, and administration of neutralizing anti-PD-L1 antibodies prevented their intrahepatic T-cell deletion. Older (≥10 months) knockouts, however, showed intrahepatic accumulation of cytotoxic CD8⁺ T cells with downregulated PD-1 and diminished apoptosis. In-vitro DNA demethylation with 5-aza-2′-deoxycytidine partially reverted PD-1 downregulation of intrahepatic CD8⁺ T cells from aged knockouts. Conclusion: Early in life, AE2 deficiency results in intrahepatic T-cell activation and PD-1/PD-L1 mediated deletion. With aging, intrahepatic CD8⁺ T cells epigenetically suppress PD-1, and their consequential expansion and further activation favor autoimmune cholangitis.     

MuRF-1 and Atrogin-1 Protein Expression and Quadriceps Fiber Size and Muscle Mass in Stable Patients with COPD

Abstract

Introduction: Animal studies demonstrate the importance of the E3 ubiquitin ligases, Muscle RING-Finger Protein 1 (MuRF-1) and atrogin-1, in muscle protein degradation during acute muscle atrophy. Small clinical studies suggest MuRF-1 and atrogin-1 expression in the quadriceps muscle is also increased in stable patients with Chronic Obstructive Pulmonary Disease compared to controls. However, it remains unclear whether these ligases have a role in maintaining a muscle-wasted state in COPD patients. Methods: 32 stable COPD patients (16 with a low fat-free mass index (FFMI), 16 with a normal FFMI) and 15 controls underwent lung function and quadriceps strength tests and a percutaneous quadriceps biopsy. Quadriceps MuRF-1 and atrogin-1 protein were quantified with western blotting. Quadriceps fiber cross-sectional area (CSA) and fiber proportions were determined by immunohistochemistry on muscle sections. MuRF-1 and atrogin-1 levels were compared between COPD patients with and without a low FFMI, and between patients and controls, and correlations between MuRF-1 and atrogin-1 levels and quadriceps fiber CSA in the patients were investigated. Results: Atrogin-1 protein levels were lower in patients than controls, but similar in patients with a low and normal FFMI. MuRF-1 levels did not differ between any groups. MuRF-1 and atrogin-1 levels were not associated with quadriceps fiber CSA or quadriceps strength in patients. Conclusions: Chronic upregulation of ubiquitin ligases was not evident in the quadriceps muscle of stable COPD patients with a low muscle mass. This does not exclude the possibility of transient increases in ubiquitin ligases during acute catabolic episodes.     

激光诱导破碎细菌的研究

本文报道了激光诱导破碎细菌的研究。破碎细菌在分子生物工程及病毒学等方面都具有极其重要的意义。用激光手段进行破碎细菌,方法简便,破碎率高。本文详述应用四种不同激光器,配合某种装置,使破碎率最高可达90%。文中对激光破碎细菌的机理作了扼要的论述。     

拉斐尔呼吸机的保养维护和常见故障排除

拉斐尔呼吸机是一台适用于儿童、成人兼用型呼吸机，该机功能齐、操作方便，满足了临床的广泛需求。本文简单介绍了该呼吸机的特点．以及日常保养维护方法和常见故障的分析、排除。     

Experimental fluid percussion brain injury: vascular disruption and neuronal and glial alterations

Because of the potential relationship between vascular disturbances and secondary tissue damage, we identified areas of brain which exhibited hemorrhage and leakage of protein during the acute stage after experimental brain injury and subsequently studied the development of pathologic changes, including cavity formation, neuronal necrosis, and gliosis within these regions. The development of pathologic changes was evaluated at 1, 6, and 24 h and 1, 2, and 4 weeks after lateral, fluid percussion (FP) brain injury of moderate severity in the rat. Vascular disruption in the acute stages, as evidenced by hemorrhage and leakage of Evans blue albumin, was most prominent 6 h postinjury and was maximal in the parieto-occipital cortex. From 1 to 24 h after injury, regions of the injured hemisphere, including the cortex and hippocampus, exhibited abnormal neurons which stained with acid fuchsin and Alizarin red, histochemical markers for injured neurons and calcium, respectively. These same regions suffered significant neuronal cell loss from 1 to 4 weeks after injury. The distribution of reactive astrocytes was also evaluated by immunocytochemical localization of glial fibrillary acidic protein (GFAP). By 2 weeks postinjury, a prominent cavity was present in the frontopariental and occipital cortices. Although astrogliosis was most pronounced in the cortex surrounding the cavity, prominent reactive astrocytes were widely distributed throughout the injured hemisphere. This study characterized the pathological changes which occur after experimental traumatic brain injury. In particular, we propose that neuronal cell injury in the hippocampus serves as a useful ‘window’ to assess beneficial efficacy of pharmacological intervention in the treatment of brain injury.     

PREDICTION AND PREVENTION OF FISTULAE AFTER MAJOR HEAD AND NECK SURGERY A PRELIMINARY REPORT

A method of checking the integrity of mucosal repair after head and neck surgery using wound amylase levels is described. A low wound amylase concentration allows early resumption of oral intake; an increasing amylase concentration is a strong predictor of subsequent wound breakdown and fistula formation. A regimen has been developed which has prevented fistulae from developing in the small number of patients studied thus far.     

Picosecond optical breakdown: tissue effects and reduction of collateral damage

The effects of picosecond laser-induced optical breakdown on tissue were investigated using high-intensity 40 ps Nd:YAG laser pulses at 1.06 microns. Tissue damage was evaluated using the corneal endothelium in vitro as a model system. Systematic studies were performed to determine the scaling of the tissue damage and damage range with pulse energy. For suprathreshold lesions, the radius of the damage zone varies as the cube root of the pulse energy, in agreement with simple physical scaling laws. A minimum damage range of less than 100 microns was observed for pulse energies of 8 muJ. Damage morphology was investigated by scanning electron microscopy. Three different damage patterns were observed; cell damage, cell removal, and rupture of Descemet's membrane. Different irradiation geometries were used to study damage mediated by either the shock wave or the cavitation bubble. Comparative studies using 10 ns pulses demonstrated that picosecond pulses yielded a significant reduction in collateral tissue damage.     

Complement Activation During Low-Density Lipoprotein Apheresis

Complement system activation was investigated in two girls with familial homozygous hypercholesterolemia undergoing two monthly sessions on LA15 or LA40 (Kaneka liposorber). We determined blood levels of C3c and C3a, leukocyte counts, and plasma levels of C3c and C3a in the extracorporeal circulation device at the start of the sessions and 15 and either 60 or 120 min into them. Sequential eluates were collected from LA40 at the end of the sessions (0.5M NaCl, 1M hydroxylamine). Anaphylatoxin C3a increased throughout, especially with LA40. As previously reported, C3a was trapped in the dextran column but was noticeably present in efferent plasma. Besides many proteins, nonnative complement fragments bearing C3a and C3d antigens were detected in almost all the eluates, suggesting possible in situ complement activation. Practically, complement activation induced by the first filter is a risk; long-term side effects may arise from this extracorporeal circulation device.     

Laser-induced fluorescence used in localizing atherosclerotic lesions

We have investigated laser-induced fluorescence frompost mortem human arteries in order to find spectroscopic properties allowing discrimination between normal and atherosclerotic vessel wall. A pulsed nitrogen laser emitting light at a wavelength of 337.1 nm was used as an excitation source. The fluorescence spectrum from 370 to 700 nm was captured and analysed by an optical multichannel analyser. Dimensionless contrast functions were formed by using characteristic spectral features at 390, 415, 480, 580 and 600 nm. All samples were investigated in scans across a region where normal as well as diseased vessel wall appeared. The types of plaque were histopathologically divided into four groups, of which three could be singled out using one or more of our spectroscopic criteria. We also investigated the different layers of the normal and diseased vessel wall in order to determine the various contributions to the fluorescence signal. Furthermore, plasma emission spectra were recorded while ablating the normal as well as the diseased vessel wall with an excimer laser, emitting radiation at 308 nm, thus detecting the change in spectral characteristics during the ablation process down into deeper layers.