Brain protection at the blood-cerebrospinal fluid interface involves a glutathione-dependent metabolic barrier mechanism.

The choroid plexuses (CPs) form a protective interface between the blood and the ventricular cerebrospinal fluid (CSF). To probe into the pathways by which CPs provide brain protection, we sought to evaluate the efficiency of glutathione conjugation in this barrier as a mechanism to prevent the entry of blood-borne electrophilic, potentially toxic compounds into the CSF, and we investigated the fate of the resulting metabolites. Rat CPs, as well as human CPs from both fetal and adult brains, displayed high glutathione-S-transferase activities. Using an in vitro model of the blood-CSF barrier consisting of choroidal epithelial cells cultured in a two-chambered device, we showed that glutathione conjugation can efficiently prevent the entry of 1-chloro-2,4-dinitrobenzene (CDNB) into the CSF, a model for electrophilic compounds. The duration of this enzymatic protection was set by the concentration of CDNB to which the epithelium was exposed, and this barrier effect was impaired only on severe epithelial intracellular glutathione and cysteine depletion. The conjugate was excreted from the choroidal cells in a polarized manner, mostly at the blood-facing membrane, via a high-capacity transport process, which is not a rate-limiting step in this detoxification pathway, and which may involve transporters of the ATP-binding cassette c(Abcc) and/or solute carrier 21 (Slc21) families. Supplying the choroidal epithelium at the blood-facing membrane with a therapeutically relevant concentration of N-acetylcysteine sustained this neuroprotective effect. Thus, glutathione conjugation at the CP epithelium coupled with the basolateral efflux of the resulting metabolites form an efficient blood-CSF enzymatic barrier, which can be enhanced by pharmacologically increasing glutathione synthesis within the epithelial cells.     

犬海水中火器伤后早期血流动力学变化

OBJECTIVE: To investigate the hemodynamic changes in the early stages of gunshot wound of dogs in seawater for exploring early treatment protocol. METHOD: Fourteen conventional Beagles models undergoing gunshot wound in seawater were used along with another 2 dogs receiving the wound on land to serve as control. After the dogs were rescued from the seawater, the respiration (R), heart rate (HR), systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial pressure (MAP), central venous pressure (CVP), pulmonary arterial wedge pressure (PAWP), and cardiac output (CO) were measured continuously in the early stages of the wound (53.62+/-12.19 min following injury), followed by statistical analysis of the results. RESULTS: Compared with the control group, the hemodynamic disturbance of the dogs receiving the wound in seawater was relatively severe during the first 15 min of the wound. The mortality tended to descend relevant to the position of the wounds, in the order of the head, chest, abdomen and limbs. CONCLUSIONS: Gunshot wound in seawater may cause severe hemodynamic changes, resulting in progressive dysfunction of circulation and high mortality rate. Early treatment should be targeted at hemodynamic stabilization in accordance to the characteristic changes during the early stages of the wound.     

恶性胸膜间皮瘤：3例尸解病例的组织化学和免疫组织化学研究

本文报告8例恶性胸膜间皮瘤尸解病例,上皮型2例,1例纤维型大体改变为巨大结节,并推挤膈肌至脐平面。对此3例进行了PAS、D-PAS,CM、AB、H-AB、CI、H-CI和CEA、Keratin、Vimentin等组化和免疫组化研究,并结合文献复习对人体胸膜间皮的组化和免疫组化特征进行了讨论,提出了人体胸膜间皮瘤的临床病理诊断与鉴别诊断意见。本组3例均合并DIC,推测本病有产生DIC的倾向性,临床对此病应警惕DIC发生的危险。     

放疗同期卡莫氟化疗治疗局部中晚期鼻咽癌的Ⅲ期临床观察

目的:探讨患者对联合应用诱导化疗和放疗同期口服卡莫氟治疗局部中晚期鼻咽癌疗效和不良反应.方法:收治66例局部中晚期鼻咽癌患者,随机分为两组,诱导化疗随后放疗组简称为对照纽,诱导化疗随后放疗同期卡莫氟治疗组简称为治疗组.两组患者数均为33例.进行根治性放疗前两组患者均接受两个疗程诱导化疗.治疗组在放疗期间同时口服卡莫氟.结果:诱导化疗有效率高,平均有效率高达87%,其中平均完全缓解率(CR)为8.7%、不完全缓解率(PR)为78.4%.对照组鼻咽肿瘤、颈部转移淋巴结全消平均剂量均高于治疗组,统计学有差异.放疗后3个月治疗组鼻咽、颈部转移淋巴结CR均高于对照组(p=0.016,P=0.042).随访期内治疗组患者局部复发率和远处转移率有降低趋势.临床治疗中出现的主要毒副反应为白细胞下降、口腔粘膜炎、急性放射性皮炎及由卡莫氟引起的热感、尿频、头昏等.结论:放疗同期口服卡莫氟治疗局部中晚期鼻咽癌,降低了鼻咽和颈部肿瘤完全消退的照射剂量,提高了鼻咽肿瘤和颈部转移灶的完全缓解率,局部复发率和远处转移率有下降趋势.尽管加卡莫氟患者毒的副作用加重,但可以耐受,经对症处理绝大部分患者能按计划完成治疗.在鼻咽癌同步化放疗方案中卡莫氟的推荐剂量为600mg/d,分3次口服.放疗同期口服卡莫氟治疗的远期疗效有待于进一步观察.     

结核性淋巴结炎的“结节病”样变型

对１２例经病理学专家会诊、从病变上认定的淋巴结“结节病”石蜡包埋组织，应用结核杆菌ＤＮＡ特异性序列片段的聚合酶链反应（Ｍ．ＴＢ－ＰＣＲ）技术、ＢＣＧ免疫组化（ＢＣＧ－ＩＨＣ）技术和抗酸染色（ＡＦ）进行了分支杆菌／结核杆菌检测。在这１２例考虑为“结节病”的病例中：有１例呈ＢＣＧ－ＩＨＣ和Ｍ。ＴＢ－ＰＣＲ两项阳性；另１例呈ＡＦ、ＢＣＧ－ＩＨＣ和Ｍ．ＴＢ－ＰＣＲ三项阳性。研究结果提示：（１）某些结核性淋巴结炎可呈结节病样病变；（２）淋巴结结节病很可能与分支杆菌／结核杆菌感染有关。     

A-raf oncogene localizes on mouse X chromosome to region some 10–17 centimorgans proximal to hypoxanthine phosphoribosyltransferase gene

The localization of the A-rafcellular oncogene on the mouse X chromosome has been determined using Xbal-restricted DNAs prepared from progeny of an interspecies backcross between the B6.CBA.R1 and the Spe/Pas mouse strains. This localization to the proximal part of the mouse X chromosome has been confirmed by the use of somatic cell hybrids, carrying partially deleted X chromosomes and suggests that the A-raf oncogene localizes to a region lying some 10–17 centimorgans proximal to the hypoxanthine phosphoribosyltransferase (Hprt) gene between the locus DXPas4and the locus DXPas7defined by the cross-reacting human X chromosome-specific probe DXS32 (M2C). This localization on the mouse X chromosome is compatible with the presence of the A-rafoncogene on the short arm of the human X chromosome between the centromere and Xp21.     

High-throughput cytochrome P450 (CYP) inhibition screening via cassette probe-dosing strategy. II. Validation of a direct injection/on-line guard cartridge extraction-tandem mass spectrometry method for CYP2D6 inhibition assessment

A highly efficient direct injection/on-line guard cartridge extraction-tandem mass spectrometry (DI/GCE-MS-MS) method has been validated for high-throughput evaluation of cytochrome P450 (CYP) 2D6 inhibition potential using human hepatic microsomes and 96-well microtiter plates. Microsomal incubations were terminated with formic acid, centrifuged, and the resulting supernatants were injected for DI/GCE-MS-MS analysis. Due to the novel use of an extremely short C18 guard cartridge, this method exhibits several advantages, such as no sample preparation, excellent on-line extraction, short run time (2.5 min), and minimized source contamination and performance deterioration. The DI/GCE-MS-MS method demonstrates acceptable accuracy and precision for the quantification of dextrorphan, a marker metabolite of dextromethorphan mediated by CYP2D6, in microsomal incubations. The CYP2D6 inhibition assay has been validated using quinidine as a known selective inhibitor of the isoform. The IC50 value (0.20 microM) measured by the new method is in good agreement with the literature value (0.22 microM).     

Fast,persistent, Ca2+-dependent K+ current controls graded electrical activity in crayfish muscle

The early outward current in opener muscle fibres of crayfish (Procambarus clarkii) was studied using the two-electrode voltage-clamp technique. This current was abolished in Ca²⁺-free and 5 mM Cd²⁺ solutions, and was blocked by extra- or intracellular tetraethylammonium, indicating that it was a Ca²⁺-dependent K⁺ current [I _K(Ca)]. I _K(Ca) was voltage dependent, apamin insensitive and sensitive to charybdotoxin (CTX), which, in addition to its tetraethylammonium sensitivity, suggests that the channels mediating I _K(Ca) behave in a BK type manner. I _K(Ca) activation was extremely fast, reaching a maximum within 5 ms, and the inactivation was incomplete, stabilizing at a persistent steady-state. I _K(Ca) was insensitive to intracellular ethylenebis(oxonitrilo)tetraacetate (EGTA), but was abolished by injection of the faster Ca²⁺ chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N,N'-tetraacetic acid (BAPTA), suggesting that voltage-dependent Ca²⁺ channels and those mediating I _K(Ca) should be clustered closely on the membrane. Under two-electrode current-clamp recording mode, low amplitude, graded responses were evoked under control conditions, whereas repetitive all-or-none spikes were elicited by application of CTX or after loading the cells with BAPTA. We conclude that I _K(Ca) activates extremely quickly, is persistent and is responsible for the generation and control of the low amplitude, graded, active responses of opener muscle fibres.