不同直径脑窥镜行微创颅内血肿碎吸术的效果比较

目的观察改良微创直视颅内血肿碎吸术与未改良者救治高血压性脑出血脑疝(HIHE)的死亡率、致残率及对脑温的影响比较。方法61例患者,随机分为两组:治疗组31例,改良NK-1纤维脑窥镜(窥镜及螺旋棒直径=4mm)行微创直视颅内血肿碎吸术,抽吸血肿70%～80%,生理盐水冲洗,腔内注入尿激酶150U,置入引流管,术后检测脑温,点式温度计的放置时间为4～5d;对照组30例,仍用NK-1纤维脑窥镜(窥镜及螺旋棒直径=6mm)抽吸,治疗方法及点式温度计放置同治疗组。两组术后均以脱水、支持疗法为主。以两组治疗结束为统计界限。以两组病死率、再出血、生存者神经功能缺损(NDS)与日常生活能力评定(ADL)、脑温为观察指标。结果治疗组病死率显著低于对照组(P<0.05);两组生存者NDS与ADL比较治疗组降低更明显(P<0.01);两组比较治疗组脑温明显降低(P<0.05);两组再出血率比较差异显著(P<0.05)。结论改良微创直视颅内血肿碎吸术能降低HIHE病死率、再出血率,提高生存者NDS和ADL,是救治HIHE较理想的方法。     

还原型谷胱甘肽稳态在腭裂发生中的作用

目的： 观察还原型谷胱甘肽稳态的改变对腭中嵴上皮细胞形态及腭裂发生率的影响,探讨氧化自由基在四氯二苯二噁英（TCDD）致畸过程中的作用。方法： 将GD10的SPF级C57BL/6J孕鼠随机分为4组,TCDD组：腹腔注射生理盐水和TCDD灌胃;TCDD+丁硫氨酸-亚砜胺(BSO)组：腹腔注射BSO 4 h后给予TCDD灌胃;BSO组：腹腔注射BSO和玉米油灌胃;对照组：腹腔注射生理盐水和玉米油灌胃。光学显微镜和电子扫描电镜下观察胎鼠腭胚突的发育及细胞表面形态。统计各组小鼠在GD17的腭裂发生率。免疫组织化学方法检测各组腭中嵴上皮各时期CYP1A1蛋白的表达。采用SPSS13.0软件包对数据进行统计分析。结果： TCDD成功构建高致畸率的腭裂模型。BSO增加了5%的腭裂发生率,但是相对于TCDD组无显著差异(P>0.05)。TCDD组腭胚突上抬时间较对照组推迟1 d,添加BSO的TCDD组腭胚突上抬过程较正常组减慢。电镜下胚胎发育的各发育时期,腭中嵴上皮表面TCDD组与对照组有显著差异。免疫组织化学检测发现,TCDD组腭中嵴上皮可见CYP1A1高表达。结论： 扰乱体内还原型谷胱甘肽稳态,可能会减少对小鼠体内氧化自由基的消耗,从而影响腭胚突的融合,继而加速了TCDD的毒性,影响腭裂发生。     

CYP2E1 induction leads to oxidative stress and cytotoxicity in glutathione-depleted cerebellar granule neurons

Increasing evidence suggests that brain cytochrome P450 (CYP) can contribute to the in situ metabolism of xenobiotics. In the liver, some xenobiotics can be metabolized by CYPs into more reactive products that can damage hepatocytes and induce cell death. In addition, normal CYP activity may produce reactive oxygen species (ROS) that contribute to cell damage through oxidative mechanisms. CYP2E1 is a CYP isoform that can generate ROS leading to cytotoxicity in multiple tissue types. The aim of this study was to determine whether CYP2E1 induction may lead to significant brain cell impairment. Immunological analysis revealed that exposure of primary cerebellar granule neuronal cultures to the CYP inducer isoniazid, increased CYP2E1 expression. In the presence of buthionine sulfoximine, an agent that reduces glutathione levels, isoniazid treatment also resulted in reactive oxygen species (ROS) production, DNA oxidation and cell death. These effects were attenuated by simultaneous exposure to diallyl sulfide, a CYP2E1 inhibitor, or to a mimetic of superoxide dismutase/catalase, (Euka). These results suggest that in cases of reduced antioxidant levels, the induction of brain CYP2E1 could represent a risk of in situ neuronal damage.     

Pro-oxidative toxicity of cells in suspension does not point to a cell cultural artefact as an explanation of the increased susceptibility of alveolar epithelial-like cells after glucocorticoid pretreatment

The influence of cell numbers on peroxide-(tertiary butylhydroperoxide (tBHP) or hydrogen peroxide-(HP)) or zinc-(zinc chloride) induced oxidative stress was assessed in alveolar epithelial-like cell lines in this work. Differences in cell numbers change the cellular glutathione and glutathione reductase activity as well as the amount of exported glutathione and therefore might influence susceptibility against oxidative stress.Toxicity due to zinc decreased, toxicity due to HP increased, while tBHP-mediated toxicity was unchanged in our experiments when cells were exposed in suspension as compared to monolayers. Toxicity of HP correlated to the glutathione content in monolayers and in cell suspensions, while zinc- or tBHP-mediated toxicity did not correlate towards glutathione.Decreasing cellular glutathione and the activity of some antioxidative enzymes by glucocorticoid pretreatment had no effect on toxicity of zinc or tBHP in L2 cells in suspensions, while toxicity in monolayers was increased. Glucocorticoid pretreatment seems to increase toxicity of HP in A549 monolayers according to the lowered protein content, while toxicity might be changed by a different way when cells are incubated as cell suspensions. No explanation as a cell culture artificial effect was observed, therefore we assume the increased toxicity after glucocorticoid pretreatment occurs in vivo as well.     

Arsenic induces telomerase expression and maintains telomere length in human cord blood cells

Inorganic arsenic (iAs) is a human carcinogen, well known as a clastogenic compound. To evaluate the molecular mechanism of arsenite (iAs^III) toxicity, we investigated the effects on cell growth and apoptosis, telomere length, telomerase expression, as well as the formation of reactive oxygen species (ROS) in male and female human cord blood cells in vitro. Incubation with iAs^III at the concentration of 0.0001 μM increased telomerase mRNA and protein expression maintained both telomere length and cellular growth, and induced mRNA over-expression of the two oncogenes ras and myc. Our results suggest that female cord blood cells are more sensitive than male ones to iAs^III induced telomerase stimulation at low concentrations, possibly related to the increased expression of ras and myc oncogenes. On the contrary, at the concentration of 1 μM, iAs^III decreased telomerase expression and telomere length, and induced apoptosis, necrosis and production of reactive oxygen species. Buthionine sulfoximine (BSO), an inhibitor of glutathione (GSH) synthesis, markedly increased the percentage of apoptotic cells, suggesting that GSH is fundamental for detoxification of iAs^III in cord blood cells. The reactive oxygen species (ROS) scavenger, 5,5-dimethyl-1-pyrroline-N-oxide (DMPO), protected cord blood cells from iAs^III toxicity, and prevented telomere shortening and telomerase down-modulation. It can be concluded that telomerase expression and telomere length are associated with iAs^III induced cell death, via production of reactive oxygen species, as well as with iAs^III induced effects on cell differentiation processes and rate of cell growth.     

Transvaginal Natural Orifice Transluminal Endoscopic Surgery (vNOTES) for prophylactic bilateral salpingo-oophorectomy

BackgroundProphylactic bilateral salpingo-oophorectomy (BSO) is an important option for reducing the risk of developing ovarian and fallopian tube cancers in women with a hereditary ovarian cancer syndrome. Conventional laparoscopy is generally preferable since it is associated with less morbidity compared to laparotomy. Transvaginal Natural Orifice Transluminal Endoscopic Surgery (vNOTES) is an emerging surgical approach that offers several advantages over conventional laparoscopy including reduced postoperative pain, low rate of surgical site infections, fast patient recuperation and better cosmetic outcome [[1], [2], [3], [4], [5], [6]]. The objective of this video is to demonstrate a surgical technique for vNOTES BSO.MethodsThis is a Stepwise demonstration of the vNOTES for prophylactic BSO with narrated video footage. The diameter of Alexis is 7 cm and we used the GelPOINT V-path transvaginal access platform (Applied Medical, Rancho Santa Margarita, CA).ResultsA 52-year-old carrier of BRCA1 mutation. The patient was selected to be operated on via a vaginal port. The video presents some tips and tricks to aid the surgeon to perform this surgery in a safe and timely manner, using the vaginal GEL POINT system and vNOTES technique.ConclusionsvNOTES for prophylactic BSO via a vaginal port is a feasible technique with promising cosmetic results. This technique allows surgeon to expose the ureter well and lower the risk of ureteric injury. Additionally, this approach avoids abdominal wall vessels injury associated with the trocar insertion. Currently, as a result of certain technical limitations, such as when performing BSO without a hysterectomy, there has been a tendency to employ the vaginal access less frequently. In this video presentation, we demonstrate the feasibility of laparoscopic BSO via vNOTES whilst leaving the uterus intact.     

Hypoxia modulates the effect of dihydroartemisinin on endothelial cells

Artemisinin derivatives, the current cornerstone of malaria treatment, possess also anti-angiogenic and anti-tumor activity. Hypoxia plays a crucial role both in severe malaria (as a consequence of the cytoadherence of infected erythrocytes to the microvasculature) and in cancer (due to the restricted blood supply in the growing tumor mass). However, the consequences of hypoxia onto the effects of artemisinins is under-researched.This study aimed at assessing how the inhibition of microvascular endothelial cell (HMEC-1) growth induced by dihydroartemisinin (DHA, an antimalarial drug and the active metabolite of currently in-use artemisinins) is affected by oxygen tension.Low doses of DHA (achieved in the patients’ plasma when treating malaria) were more inhibitory in hypoxia, whereas high doses (required for anti-angiogenic or anti-tumor activity) were more effective in normoxia. The peroxide bridge is essential for cellular toxicity (deoxyDHA was inactive). High doses of DHA caused HMEC-1 apoptosis and G2 cell cycle arrest. Effects were mediated by the generation of oxidative stress as demonstrated by DCF-DA fluorescence and membrane lipid peroxidation analysis.Overall, these results suggest that DHA inhibition of endothelial cell growth is related to the level of tissue oxygenation and drug concentration. This should be considered when studying both the effects of artemisinin derivatives as antimalarials and the potential therapeutic applications of these drugs as anti-tumor agents.     

Role of protein haptenation in triggering maturation events in the dendritic cell surrogate cell line THP-1

Dendritic cell (DC) maturation in response to contact sensitizers is a crucial step in the induction of sensitization reactions; however the underlying mechanism of activation remains unknown. To test whether the extent of protein haptenation is a determinant in DC maturation, we tested the effect of five dinitrophenyl (DNP) analogues of different reactivity, on maturation markers in the cell line, THP-1. The potencies of the test compounds in upregulating CD54 levels, inducing IL-8 release and triggering p38 MAPK phosphorylation did not correlate with their ability to deplete intracellular glutathione (GSH) levels or cause cell toxicity. However, the compounds' potency at inducing p38 phosphorylation was significantly associated with the amount of intracellular protein adducts formed (p < 0.05). Inhibition experiments show that, at least for DNFB, p38 MAP kinase signalling controls compound-specific changes in CD54 expression and IL-8 release. 2D-PAGE analysis revealed that all the DNP analogues appeared to bind similar proteins. The analogues failed to activate NFkB, however, they activated Nrf2, which was used as a marker of oxidative stress. Neither GSH depletion, by use of buthionine sulfoximine, nor treatment with the strongly lysine-reactive hapten penicillin elicited maturation. We conclude that protein haptenation, probably through reactive cysteine residues may be a trigger for maturation events in this in vitro model and that p38 activation may be a discriminatory marker for the classification of potency of chemical sensitizers.