氯胺酮对氯化钾诱发豚鼠耳蜗外毛细胞钙离子内流的影响

目的 评价氯胺酮对氯化钾诱发豚鼠耳蜗外毛细胞钙离子内流的影响,探讨其对听觉外周感受器(耳蜗)的作用.方法 成年豚鼠12只,雌雄不拘,迅速断头处死,暴露耳蜗,取出双侧听泡,采用酶孵育后,机械分离法分离外毛细胞.选择30个活力较高的外毛细胞,随机分为3组(n=10):对照组(C组)、100 μmol/L氯胺酮组(K1组)和200 μmol/L氯胺酮组(K2组).用Fluo-3AM钙荧光指示剂染色后,K1组和K2组分别给予100、200 μmol/L氯胺酮处理5 min,然后加入40 mmol/L氯化钾,采用激光共聚焦显微镜测定细胞内荧光强度,反映细胞内游离钙离子浓度([Ca2+]i).结果 与基础值比较,各组加入氯胺酮后[Ca2+]i差异无统计学意义(P＞0.05),加入氯化钾后[Ca2+]i升高(P＜0.01);K1组和K2组加入氯化钾后[Ca2+]i低于C组,且K2组低于K1组(P＜0.05或0.01).结论 氯胺酮可抑制豚鼠耳蜗外毛细胞Ca2+跨膜内流,从而降低[Ca2+]i.     

异丙酚对大鼠心肌细胞钙离子移动及钙通道电流的影响

目的 观察异丙酚对心肌细胞钙离子移动和Ｌ－型钙通道电流（Ｉｃａｌ）的影响。方法 急性分离的大鼠单个心室肌细胞，采用Ｆｌｕｏ－３ＡＭ钙荧光指示剂和膜片钳技术，观察５０μｍｏｌ．Ｌ＾－１或２５０μｍｏｌ．Ｌ＾－１异丙酚对高浓度ＫＣ１或咖啡因所诱发细胞内钙离子浓度的变化，以及对Ｌ－型钙通道电流的影响。结果 ５０μｍｏｌ．Ｌ＾－１或２５０μｍｏｌ．Ｌ＾－１异丙酚明显减弱ＫＣｌ所诱发细胞内钙离子的增加，浓度     

P物质对体外培养的胃癌细胞内游离钙浓度的影响

目的研究P物质（substance P，SP）对体外培养的人低分化胃癌细胞MKN45内游离钙浓度的影响。方法在RPMI1640培养液中培养人胃癌细胞系MKN45，应用钙特异性荧光指示剂Furu-3／AM负载细胞，用ASN-1377642（NK-1受体拮抗剂）、尼卡地平和不同浓度的SP作用于人胃癌细胞MKN45，采用激光扫描共聚焦显微镜观测胃癌细胞内游离钙浓度的变化。结果在Hanks液（含钙离子）中加入10、50及100nmol／LSP可以显著升高胃癌细胞内钙离子浓度（P〈0．05），并且呈剂量依赖性（P〈0．05）；在无外钙时，SP引起细胞内钙离子浓度升高的幅度低于有外钙时（P〈0．05）；在Hanks液中加入NK-1受体拮抗剂ASN-1377642和钙通道阻滞剂尼卡地平孵育后，SP引起胃癌细胞内钙离子浓度升高的幅度显著降低，与Hanks液组比较差异有统计学意义（P〈0．05）。结论SP可以显著升高胃癌细胞内钙离子浓度，细胞内钙离子浓度的升高源于内钙释放和外钙内流。     

七氟醚对豚鼠耳蜗外毛细胞Ca2+跨膜内流和内质网钙释放功能的影响

目的 观察七氟醚对豚鼠耳蜗外毛细胞Ca2+跨膜内流和内质网钙释放功能的影响,探讨其对听觉外周感受器(耳蜗)作用的可能机制.方法 第一部分成年豚鼠,雌雄不拘,迅速断头,取耳蜗,酶孵育后,机械分离法分离外毛细胞,30个活力良好的外毛细胞随机分为3组(n=10):对照组(C组),低浓度七氟醚组(S1组)和高浓度七氟醚组(S2组),用Fluo-3AM钙荧光指示剂染色后,分别给予纯氧、1.7%七氟醚、3.4%七氟醚处理20min,然后加入40mmol/L氯化钾,测定细胞内游离钙离子浓度([Ca2+];).第二部分以20 mmol/L 咖啡因代替氯化钾其余处理及分组同第一部分.结果 第一部分与基础值比较,各组给予七氟醚后[Ca2+];差异无统计学意义(P＞0.05),加入氯化钾后[Ca2+];升高(P＜0.01);S1组和S2组加入氯化钾后[Ca2+];低于C组,且S2组低于S1组(P＜0.05).第二部分与基础值比较,各组给予七氟醚后[Ca2+];差异无统计学意义(P＞0.05),加入咖啡因后[Ca2+];升高(P＜0.0);加入咖啡因后各组[Ca2+];差异无统计学意义(P＞0.05).结论 七氟醚可浓度依赖性地抑制豚鼠耳蜗外毛细胞电压依赖型Ca2+通道开放,而对内质网Ryanodine敏感性钙释放功能无影响.     

草氨酸盐对人胰腺癌细胞内钙含量影响的研究

目的:探讨草氨酸盐(oxamate)对人胰腺癌细胞内钙含量的影响。方法:体外培养人胰腺癌细胞panc-1，用oxamate干预癌细胞后48h，行钙离子荧光指示剂Fluo-3/AM染色，激光共聚焦下观察不同浓度oxamate作用下细胞内钙含量的变化。结果:oxamate干预48h后，细胞内钙含量随着oxamate浓度的升高而致荧光强度逐渐增强，呈剂量-效应关系。结论:oxamate能增加panc-1细胞内钙离子含量，影响细胞内钙离子平衡，进而干涉细胞病理生理过程。     

三氧化二砷对小鼠膀胱癌细胞系细胞内游离钙浓度影响的实验研究

目的：研究As2O3对小鼠膀胱癌细胞系WYH929肿瘤细胞内游离钙离子浓度的影响。方法：用钙离子荧光探剂Fura2-AM,以荧光分光光度计检测As2O3对膀胱癌细胞内游离钙离子浓度的变化。结果：As2O3可以使肿瘤细胞[Ca^2 ]i水平明显升高，由88nmol/L逐渐升高至230nmol/L，存在明显的量效关系（P＜0．05），而与细胞外钙离子浓度变化无关（P＞0．05）。结论：As2O3影响小鼠膀胱癌细胞株WYH929肿瘤细胞内钙稳态，诱导肿瘤细胞内钙离子浓度持续升高；这可能是其抗肿瘤，诱导肿瘤细胞凋亡作用的机制之一。     

加味小承气颗粒剂对小鼠腹腔巨噬细胞合成白三烯B_4及对细胞内游离钙浓度的影响

目的：研究加味小承气（ＸｉａｏＣｈｅｎｇＱｉ,ＸＣＱ）颗粒剂治疗腹腔炎症机理与小（大）鼠腹腔巨噬细胞（ＭΦ）合成释放白三烯Ｂ４以及对细胞内游离钙浓度的影响。方法：应用高效液相色谱分析ＬＴＢ４,应用Ｆｕｒａ－２／ＡＭ及荧光分光光度计测定细胞内钙离子浓度。结果：不同浓度的ＸＣＱ颗粒剂（０１５～１０ｇ／Ｌ）明显抑制大鼠（ＭΦ）合成释放白三烯Ｂ４并抑制小鼠细胞内钙离子释放（静止期细胞内钙离子浓度为６３４５±１２４４ｎｍｏｌ／Ｌ,与对照组（１４８３９±１６６２ｎｍｏｌ／Ｌ）比较有显著性差别,在细胞外钙浓度增高条件下,ＸＣＱ尚促使外钙内流。结论：小承气颗粒剂抗炎机理与抑制小（大）鼠（ＭΦ）合成释放白三烯Ｂ４及调节细胞内外钙离子浓度有关。     

几种静脉麻醉药对大鼠心室肌细胞游离钙离子移动的影响

目的　观察临床常用静脉麻醉药咪唑安定、异丙酚和依托咪酯对心肌细胞内钙离子移动的作用 ,探讨其对心肌收缩性的影响。方法　急性分离的大鼠单个心室肌细胞 ,采用 Fluo- 3 / AM为钙荧光指示剂 ,观察临床诱导峰浓度及 5倍峰浓度时咪唑安定 ( 3μm ol· L- 1 或 15μmol· L- 1 )、异丙酚( 5 0μm ol· L- 1 或 2 5 0μmol· L- 1 )或依托咪酯 ( 3μmol· L- 1 或 15μmol· L- 1 )对高浓度 KCl或咖啡因所诱发细胞内钙离子浓度变化的影响。结果　临床诱导峰浓度异丙酚明显减弱 KCl所诱发的钙离子跨膜内流 ( P<0 .0 5 ) ,而咪唑安定和依托咪酯 ( 3 μm ol· L- 1 )有减弱钙离子跨膜内流趋势 ;而超临床浓度的咪唑安定、异丙酚和依托咪酯明显减弱 KCl所诱发钙离子跨膜内流 ( P<0 .0 5 )。上述两种浓度咪唑安定、异丙酚或依托咪酯对咖啡因所致胞浆内钙离子的增加均无明显影响 ( P>0 .0 5 )。结论　上述几种静脉麻醉药具有剂量依赖性地减少心肌细胞钙离子跨膜内流作用 ,以异丙酚影响最显著 ,咪唑安定次之 ,依托咪酯较弱 ,但对心肌细胞肌浆网功能尚无明显影响     

异丙酚对中性粒细胞活性氧生成及细胞内游离Ca~(2 )浓度的影响

目的 :测定异丙酚对中性粒细胞 (PMN)激活后活性氧生成以及细胞内游离Ca2 浓度 ([Ca2 ]i)的影响 ,探讨异丙酚影响PMN呼吸爆发的内在机制。方法 :利用电子自旋共振 (ESR)技术和自旋捕捉法 ,观察异丙酚对PMN呼吸爆发产生氧自由基 (ROS)的抑制作用 ;同时采用钙荧光指示剂 (Fura 2 /AM)负载离体PMN并行双波长模式测定不同浓度异丙酚对静息和激活状态下PMN细胞内 [Ca2 ]i 的影响。结果 :高浓度 (75 μM)异丙酚可使静息状态下PMN细胞内 [Ca2 ]i升高 ;在有钙基质中 ,异丙酚对趋化三肽 (fMLP)诱发的PMN胞内 [Ca2 ]i 的升高有明显的降调作用 ,而在无钙基质中则不明显。在无钙基质中 ,高浓度 (≥ 5 0 μM )的异丙酚对fMLP诱发PMN呼吸爆发产生ROS有明显的抑制作用 ;而存在胞外钙浓度时 ,2 5 μM异丙酚的抑制程度更为明显。结论 :异丙酚可通过作用于胞外钙内流和胞内储存钙释放影响静息及激活状态下PMN胞内 [Ca2 ]i;异丙酚在有钙基质中对PMN呼吸爆发产生ROS的抑制作用更为明显 ,提示异丙酚影响PMN胞内 [Ca2 ]i,特别是对胞外钙内流的抑制作用 ,可能是其抑制PMN呼吸爆发的内在机制之一。     

NO与cGMP协同降低细胞内钙离子的释放从而使阴茎勃起

阴茎海绵体平滑肌细胞（SMCs）强直性收缩使阴茎处于疲软状态，NO可使SMCs舒张，从而使阴茎勃起。Williams等研究了SMCs内NO受肾上腺素能刺激后的作用。从小鼠和人类分离新鲜的SMCs，利用Fura-2荧光标记法，测量细胞内钙离子浓度（[Ca^2＋]（i））。结果显示，不论细胞外是否存在钙离子，苯肾上腺素（PE）均能短暂性提高细胞内钙离子浓度，表明细胞内钙离子库有释放活动。     

2015年乳腺癌辅助化疗进展

【摘要】〓乳腺癌是危害我国女性健康的头号杀手,尽管近年来辅助化疗的研究进展突飞猛进,但临床中仍有不少问题未能明确,如辅助化疗的合适人群、化疗的开始时间、蒽环及紫杉类的地位和用法、强化维持治疗的作用、疗效及预后的生物标志物等。本文结合乳腺癌辅助化疗在临床上的常见问题和2015年各大乳腺癌会议阐述乳腺癌辅助化疗的最新进展。     

Alterations in T-Cell Subset Frequency in Peripheral Blood in Obesity

Background: Obesity affects the regulation of immune and inflammatory responses. This study characterizes differences in peripheral blood lymphocyte phenotype in obese humans. Methods: Frequencies of lymphocyte subsets among peripheral blood mononuclear cells were compared between 10 obese (BMI ≥35) and 10 lean subjects, as determined by antibodies directed against cluster differentiation (CD) markers. Results: Obese patients demonstrated an increased frequency of CD3+CD4+ T-cells (mean difference 12%, P=0.004), a decreased frequency of CD3+CD8+ T-cells (mean difference 9.4%, P=0.016) and an increased frequency of CD3+CD8+CD95+ T-cells (mean difference 13.3%, P=0.032). No other differences among T-cell or monocyte subsets were noted. Conclusions: Obesity is associated with alterations in frequencies of peripheral CD4+ and CD8+ T-cells and aberrations in the expression of CD95 among CD8+ T-cells. These data suggest both CD4+ and CD8+ T-cell compartments, as well as the regulation of CD95 expression on CD8+ T-cells, as targets for further study into obesity's effects on the immune system.     

西宁地区烧伤病人动脉血气分析观察

对高海拔地区的27例烧伤病人动脉血气变化进行了分析和观察。结果证明:无论是存活病人还是死亡病人伤后均存在有低氧血症问题。并且在死亡病人和烧伤合并吸入性损伤病人其低氧血症的发生早于单纯烧伤病人。提示:吸入性损伤病人应立即行气管切开术以保障氧气供给,单纯烧伤病人可常规吸氧以维持正常血 PaO_2,ARDS 均发生在合并吸入性损伤的病人,高频喷射通气技术对纠正低氧血症有一定效果。     

Controversies in Fistula in Ano

Managing a complex fistula in ano can be a daunting task for most surgeons; largely due to the two major dreaded complications—recurrence & fecal incontinence. It is important to understand the anatomy of the anal sphincters & the aetiopathological process of the disease to provide better patient care. There are quite a few controversies associated with fistula in ano & its management, which compound the difficulty in treating fistula in ano. This article attempts to clear some of those major controversies.     

β-半乳糖苷酶在体内外成骨细胞中的表达

目的 研究β—半乳糖苷酶(β—gal)在成骨细胞中的表达状况，为阐明MorquioB综合征的发病机制提供依据。方法 裸鼠各器官和骨组织标本行X-gal染色检测。抽取羊和人骨髓行骨髓基质细胞(BMSCs)培养，分为4组：I：Adv-hBMP-2转染组；Ⅱ：Adv—β—gal转染组；Ⅲ：未转染组；Ⅳ：地塞米松诱导组。分别行X-gal染色和RT-PCR检测β—gal的表达。结果 裸鼠骺板两侧、骨膜内面及松质骨的成骨细胞和破骨细胞可见多量β—gal的表达。未转染BMSCs组有少量β—gal的表达，其他3组细胞的β—gal表达增高。结论成骨细胞和破骨细胞可表达多量β—gal，该两种细胞的β—gal缺乏可能是MorquioB综合征骨骼异常的直接原因。     

Fluid-phase transcytosis in the primate epididymis in vitro and in vivo

Ligated tubules from the corpus epididymidis of men and monkeys were incubated in medium containing horseradish peroxidase (HRP) as a marker for fluid-phase endocytosis. HRP was localized by light and electron microscopy after 0, 15, 30 and 60 min of incubation. Movement between the cells was prevented by tight junctions, but bypass of this barrier was apparently achieved by an intracellular vesicular mechanism leading to a time-dependent appearance of HRP in the lumen. Uptake of HRP into basal cells and capture by the lysosomal apparatus of principal cells were also observed. HRP-filled vesicles also appeared in the basal, mid and apical cytoplasm of epithelial cells in the caput 1 h after injection of the tracer into the epididymal circulation of the monkey, suggesting that this pathway also operates in vivo.     

Surgical management of ductal carcinoma in situ in Australia in 1995

Background: In the present paper we describe the presentation and management of ductal carcinoma in situ (DCIS) of the breast in women in Australia in 1995. This representative, national data set provides a historical comparator for studies examining DCIS management that follow. Methods: Surgeons identified by population‐based cancer registries as having treated a new diagnosis of DCIS between 1 April and 30 September 1995 completed a questionnaire on the presentation and management of each case. Results: Two hundred and five surgeons supplied treatment details on 418 DCIS tumours in 415 women . Half of all tumours were detected at BreastScreen clinics and a further 25% were detected at other mammography centres. Twenty‐six percent of tumours were palpable at presentation, 33% were multifocal and 55% were high grade (including comedocarcinoma). Breast conserving therapy (BCT) rather than mastectomy was utilized in 260 (62%) of cases. Tumours that were of low grade, small in size and not multifocal were more likely to be treated by BCT. Surgeons seeing six or more DCIS cases in the 6‐month period were more likely to utilize BCT. Of the conservatively treated cases, 22% were referred for a radiation oncology consultation. The most common reasons for treating DCIS with mastectomy were that the tumour was too extensive or multifocal (63%), it extended to margins of the specimen (42%), or patient concerns about recurrence (34%). Conclusions: In 1995 the majority of DCIS was treated with breast conserving surgery alone. Surgeons treating more DCIS cases were more likely to perform conservative surgery than surgeons treating only one DCIS case in the study period.     

Smoking-Attributable mortality in Spain in 2016

IntroductionSmoking-attributable mortality (SAM) is a valuable indicator that can be used to characterize the course and health burden of the smoking epidemic. The aim of this paper was to estimate SAM in Spain in 2016 in the population aged 35 and over, using the best available evidence.MethodsA smoking prevalence-dependent analysis based on the estimation of population-attributable fractions was performed. Smoking prevalence (never, former, and current smokers) was calculated from a combination of the Spanish Health Survey (2016) and the European Health Survey (2014); the relative risk of death among current and former smokers was taken from the follow-up of various cohorts; and mortality rates were obtained from National Center for Statistics data. SAM estimates are presented globally, and by sex, age groups, and major disease categories: cancer, cardiometabolic diseases and respiratory diseases.ResultsIn 2016, 56,124 deaths were attributed to tobacco consumption, 84% in men (47,000), and 50% in the population aged over 74 (27,795). Overall, 50% of SAM was due to cancer (28,281), 65% of which was lung cancer. One in 4 attributable deaths (13,849) occurred before the age of 65.ConclusionsOne in 7 deaths in Spain in 2016 were attributable to smoking. This estimation of SAM clearly highlights the great impact of smoking on mortality in Spain, mainly due to lung cancer and chronic obstructive pulmonary disease.