强脉冲光对长波紫外线Ⅰ诱导成纤维细胞光损伤的保护作用

目的 研究强脉冲光（intense pulse light,IPL）对长波紫外线（ultraviolet A,UVA Ⅰ）诱导的正常离体人皮肤成纤维细胞（fibroblast,FB）损伤的保护作用,探讨以IPL为手段的光子嫩肤技术的理论基础.方法 分离并培养人FB,用不同剂量UVA Ⅰ照射FB以诱导细胞光损伤,CCK-8检测其增殖能力的情况,根据预实验结果确定IPL剂量进行照射,流式细胞仪技术枪测细胞周期,Western印迹法检测CylinD1和CDK2蛋白的表达水平.结果 不同剂量的UVA Ⅰ可造成FB的损伤,随着UVA Ⅰ剂量的增加,细胞增殖能力下降,11 J/cm2的剂量可明显导致细胞大量死亡,而7 J/cm2的UVAⅠ对细胞的增殖能力没有明显的影响;经UVA Ⅰ照射2 d后再进行IPL连续照射2 d,细胞增殖活性高于单独UVA Ⅰ处理组,差异具有统计学意义.流式细胞仪检测结果表明该组细胞增殖指数升高.细胞周期蛋白CyclinD1和CDK2的表达水平升高.结论 IPL可通过调节周期蛋白的表达而促进正常FB增殖,从而保护UVAⅠ诱导的FB光损伤,为应用IPL面部美容除皱即光子嫩肤术提供理论依据.     

盐酸川芎嗪在长波紫外线诱导人皮肤成纤维细胞氧化应激中的作用

目的:探讨盐酸川芎嗪(2,3,5,6-tetramethylpyrazine,TMP)在长波紫外线(Utraviolet A,UVA)诱导人皮肤成纤维细胞(Human dermis fibroblasts,HDF)发生氧化应激反应中的保护作用。方法:原代培养HDF细胞,用不同浓度TMP分别作用未经UVA照射以及经UVA多次照射的HDF细胞24 h,48 h,72 h,CCK-8法检测TMP对HDF细胞体外增殖的作用;流式细胞术检测不同浓度TMP对多次UVA照射后HDF细胞内活性氧(reactive oxygen species,ROS)总量的影响。结果:TMP在一定浓度范围内对HDF细胞的体外增殖无明显促进或者毒性作用,但对多次UVA照射后HDF细胞的体外增殖有一定的促进作用,且与浓度有一定的相关性,与UVA对照组比较有统计学意义(P<0.05);TMP可清除UVA多次照射引起的HDF细胞内ROS累积,与UVA对照组比较有统计学差异(P<0.05),其作用效果能达未经UVA照射的水平。结论:盐酸川芎嗪(TMP)对长波紫外线诱导HDF细胞氧化应激有一定的保护作用。     

强脉冲光对成纤维细胞TGF-β1表达的影响

目的 研究强脉冲光(IPL)对皮肤成纤维细胞TGF-β1表达的影响,探索IPL影响皮肤胶原合成的调控机制.方法 将体外分离培养的人包皮原代成纤维细胞,分成照射组和对照组,照射组以波长590～1200 nm、能量密度为29 J/cm2、脉宽15 ms、延时5 ms的IPL对成纤维细胞进行照射.两组细胞均采用无血清DMEM分别培养1、12、24、48 h,然后荧光定量PCR法分别测定4个时间点细胞TGF-β1的表达情况.结果 培养12、24、48 h后,TGF-β1的表达在照射组成纤维细胞中明显增高,与对照组比较,差异有统计学意义(P<0.01).结论 IPL能促进成纤维细胞合成TGF-β1的活性.     

TGF-β1对UVA照射皮肤成纤维细胞HSP70表达的影响

目的:探讨转化生长因子-β1(tansforming gowth fetor-beta 1,TGF-β1)对长波紫外线(ultraviolet A,UVA)照射皮肤成纤维细胞热休克蛋白70(heat shock protein 70,HSP70)表达的影响.方法:通过酶联免疫法(ELISA)测定不同剂量UVA即对照组(UVA 0J/cm2)、UVA 5J/cm2、UVA 10 J/cm2、UVA 20J/cm2照射成纤维细胞上清液中HSP70的含量;选择UVA照射剂量为15J/cm2,不同剂量TGF-β1即小剂量组(UVA TGF-β1 0.1ng/m1)、中剂量组(UVA TGF-β1 1ng/m1)、大剂量组(UVA TGF-β1 10ng/m1)处理后成纤维细胞HSP70上清液中HSP70的含量.结果:UVA照射体外培养的皮肤成纤维细胞导致HSP70表达下降,UVA 20J/cm2照射组与对照组比较,HSPT0含量明显降低,有非常显著性差异(P＜0.01),OVA 10J/cm2照射组差异有统计学意义(P＜0.05);不同剂量TGF-β1处理后,大剂量TGF-β1,组,皮肤成纤维细胞上清液中HSP70含量明显升高,与照射组比较有非常显著性差异(P＜0.01),中剂量组差异有统计学意义(P＜0.05).结论:UVA照射抑制体外培养成纤维细胞HSPT0表达,TGF-β1可提高UVA照射体外培养的皮肤成纤维细胞HSP70表达水平,对皮肤成纤维细胞起保护作用.     

组织蛋白酶G在光老化皮肤成纤维细胞中的表达及意义

目的 探讨组织蛋白酶G（cathepsin G）在光老化皮肤中的表达变化及意义.方法 培养原代人皮肤成纤维细胞,8-甲氧补骨脂素＋长波紫外线（8-MOP＋UVA）法体外诱导培养细胞光老化.衰老相关-β半乳糖苷酶（SA-β-Gal）染色证明老化诱导成功.蛋白印迹技术及反转录-聚合酶链式反应（RT-PCR）对比检测光老化成纤维细胞及正常成纤维细胞组织蛋白酶G蛋白及基因表达.结果 Western印迹法结果 示光老化诱导组组织蛋白酶G表达上调为单UVA诱导组、单8-MOP孵育组及空白组的（1.70±0.028）倍.实时RT-PCR结果 示光老化细胞组织蛋白酶G mRNA表达上调为空白组的（1.42±0.09）倍（P〈0.01）.结论 组织蛋白酶G在光老化成纤维细胞中高表达,可能通过改变细胞外基质成分和激活基质金属蛋白酶参与皮肤光老化所致.     

低剂量长波紫外线诱导培养人皮肤成纤维细胞适应性反应观察

目的:探讨低剂量长波紫外线诱导培养人皮肤成纤维细胞适应性反应的程度及特点。方法:以具有致死作用的86.4J/cm2UVA照射经7.2J/cm2低剂量UVA单次或多次预照射的培养人皮肤成纤维细胞,光镜、电镜观察细胞形态学变化,流式细胞仪检测细胞凋亡的比例,单细胞凝胶电泳检测DNA损伤的程度。结果:单次或多次7.2J/cm2UVA预照射,随后86.4J/cm2UVA照射,观察其对培养皮肤成纤维细胞形态学上的毒性反应,并使细胞凋亡的比例下降、DNA链断裂减少及修复加快,和单纯86.4J/cm2UVA照射的相应细胞比较均有显著差异(P<0.01或P<0.05);单次7.2J/cm2的UVA预照射诱导培养皮肤成纤维细胞的适应性反应在预照射12h后消失,而当低剂量UVA预照射的累积剂量达到28.8J/cm2以上时,预照射的培养细胞即使是14天后对86.4J/cm2UVA照射仍有明显的防护作用。结论:低剂量UVA照射可诱导培养的皮肤成纤维细胞出现对随后高剂量UVA照射的适应性反应,其滞留期及强度与低剂量UVA的累积剂量有关。     

强脉冲光促进大鼠皮肤转化生长因子-β1表达的实验研究

目的观察强脉冲光照射对SD大鼠皮肤转化生长因子-β1(Transforming growth factor-β1,TGF-β1)表达的影响,以进一步阐明强脉冲光治疗皮肤光老化可能的机制。方法随机选取15只SD大鼠,两侧背部皮肤去毛,以一侧为强脉冲光照射实验组,对侧背部皮肤作对照组。采用波长560～1200 nm、能量27 J/cm~2、脉宽3.0 ms、双脉冲方式、脉冲延时10 ms的强脉冲光照射实验组皮肤,取术前及术后4周大鼠实验侧和对照侧皮肤标本,行大鼠皮肤TGF-β1蛋白免疫组化染色定位,观察大鼠皮肤TGF-β1的分布情况以及强脉冲光照射后TGF-β1的表达变化。结果TGF-β1分布于大鼠正常皮肤全层,主要分布在细胞外结缔组织基质胶原纤维中。强脉冲光照射刺激大鼠皮肤分泌TGF-β1,实验组强脉冲光照射4周后,大鼠皮肤TGF-β1表达较对照组显著增强(P<0.001),阳性产物主要分布在细胞外结缔组织中。真皮浅层较对照组有非常强烈的棕黄色着染,同时皮肤表皮细胞亦出现明显的阳性表达产物。阳性表达区成纤维细胞数量明显增多。结论强脉冲光照射后上调大鼠皮肤TGF-β1的表达,是强脉冲光治疗皮肤光老化的内在机制。     

沉默p53和p21基因延缓髓核细胞衰老退变实验研究

目的髓核细胞衰老凋亡是椎间盘退行性变的病理基础,探讨髓核细胞表型分子及延缓髓核细胞衰老退变的机制。方法原代培养8～10周龄雄性SD大鼠髓核细胞,免疫细胞化学染色鉴定髓核细胞表型分子低氧诱导因子1α(hypoxia inducible factor 1α,HIF-1α)、HIF-1β、基质金属蛋白酶2(matrix metalloproteinase 2,MMP-2)及Ⅱ型胶原表达。小干扰RNA(small interference RNA,siRNA)瞬时转染髓核细胞沉默p53、p21后行RT-PCR及Western blot检测沉默效率,siRNA转染前后细胞衰老相关β半乳糖苷酶(senescence associated-β-galactosidase,SA-β-gal)染色检测髓核细胞衰老变化,流式细胞仪检测髓核细胞周期变化,MTT法生长曲线分析髓核细胞增殖变化。结果免疫细胞化学染色显示髓核细胞表达HIF-1α、HIF-1β、MMP-2及Ⅱ型胶原。第35代髓核细胞转染p53、p21 siRNA后RT-PCR及Western blot检测示p53、p21表达明显受到抑制。第35代髓核细胞SA-β-gal染色阳性率明显高于第1代髓核细胞(P<0.001);正常第35代髓核细胞经p53 siRNA(p53转染组)和p21 siRNA(p21转染组)转染后,SA-β-gal阳性率明显低于正常第35代髓核细胞(正常组)和加入脂质体LipofectaminTM2000而无siRNA的第35代髓核细胞(阴性对照组),差异有统计学意义(P<0.001)。细胞周期分析显示,p53转染组及p21转染组G1期百分比均明显低于正常组和阴性对照组(P<0.05),S期百分比明显高于正常组和阴性对照组(P<0.05)。MTT生长曲线显示转染p53、p21 siRNA后可促进髓核细胞增殖。结论沉默p53、p21基因可通过调节细胞周期而抑制髓核细胞衰老退变,改善椎间盘退变过程;沉默p53、p21基因可能是潜在的治疗椎间盘退行性变的方法。     

雷帕霉素下调光老化成纤维细胞老化表型的初步研究

目的探讨雷帕霉素(RAPA)对紫外线照射导致的成纤维细胞(FBs)衰老和抗氧化应激的作用特点。方法采用CCK-8法检测不同浓度的RAPA对FBs活性的影响,甄选最佳应用浓度。利用紫外线体外反复照射构建成纤维细胞的老化模型。RAPA作用后,β-半乳糖苷酶染色法检测衰老细胞,利用DCFH-DA为荧光探针,用流式细胞仪检测细胞内活性氧簇(ROS)的水平。结果低浓度的RAPA对细胞的活性并无影响;与紫外线照射组组比较,RAPA+紫外线组的β-半乳糖苷酶染色阳性率下降了约40%,差异具有统计学意义(P0.05)。RAPA预处理可以明显降低紫外线照射细胞组ROS的产生。结论 RAPA可以降低光老化成纤维细胞ROS生成量,一定程度上缓解了紫外线照射导致的细胞光老化进程。     

年龄因素对大鼠髓核细胞生物学特性的影响

[目的]探讨年龄因素对大鼠髓核细胞(nucleus pulposus cells,NPCs)生物学特性的影响。[方法]分别从青年(3个月龄)和老年(14个月龄)雄性SD大鼠分离培养NPCs。镜下观察两组NPCs形态特征和生长状况;待细胞传至第3代时,绘制生长曲线评估其增殖能力;衰老相关β半乳糖苷酶(senescence associated-beta-galactosidase,SA-β-gal)染色法测定NPCs衰老情况;流式细胞仪检测细胞周期分布;RT-PCR检测Ⅱ型胶原、蛋白多糖及SOX-9的mRNA相对表达。[结果]两组细胞均呈短梭形或多角形,老年组NPCs胞体略偏大、扁平,胞质的折光性减弱,核变大;青年组NPCs较老年组贴壁、生长快,达到细胞融合状态所需时间短;生长曲线结果显示,进入对数生长期后,青年组NPCs增殖速率显著高于老年组;SA-β-gal染色结果显示,青年组蓝染的NPCs呈散在分布,而老年组多呈簇状分布,其SA-β-gal阳性率显著高于青年组(P0.05);细胞周期分析结果显示,老年组G1期细胞的百分比显著高于青年组(P0.05),S期细胞百分比显著低于青年组(P0.05);RT-PCR检测结果显示,青年组Ⅱ型胶原、蛋白多糖及SOX-9的mRNA相对表达量显著高于老年组(P0.05)。[结论]随年龄增长,NPCs衰老后其生物学活性及表型基因的表达降低。     

Hyperbaric Oxygen and N-Acetylcysteine Treatment in L-Arginine-Induced Acute Pancreatitis in Rats

ABSTRACT

Background: This study was designed to evaluate the combined effects of hyperbaric oxygen (HBO) and N-acetylcysteine (NAC) on acute necrotizing pancreatitis in rats. Methods: Experiments were performed in 50 male Wistar rats, which were divided into five groups (N = 10 for each group). The first group received normal saline (0.9% NaCl) intraperitoneal and served as the control group. In the second group, acute pancreatitis was induced by 3.2-g/kg body weight L-arginine intraperitoneal twice at an interval of 1 hr, which has been shown previously to produce severe necrotizing acute pancreatitis. In the third group, NAC treatment (1000 mg/kg) was given after 1 hr of the induction of acute pancreatitis twice 24 hr apart. In the fourth group, animals received HBO, 6 hr after the induction of pancreatitis twice 12 hr apart. In the fifth group, animals received together NAC as in Group 3 and HBO treatment as in Group 4. Groups 1, 2, and 3 were left under normal atmospheric pressures. Twelve hours after last treatment, the animals were killed by exsanguinations. Blood samples were studied for amylase, calcium, and lactate dehydrogenase (LDH), pancreatic histology, pancreatic tissue malondialdehyde, superoxide dismutase, and glutathione levels. Results: Acute pancreatitis is reduced by the treatment of NAC, HBO, NAC + HBO. HBO + NAC groups performed statistically the best in preventing L-arginine-induced acute necrotising pancreatitis. Conclusions: NAC especially combined with HBO, decreases oxidative stress parameters, serum amylase, calcium, and LDH levels, as well as histopathologic score.     

硝普钠阴茎海绵体内注射治疗阳萎的临床研究

本研究选择４２例阳萎患者，采用硝普钠进行阴茎海绵体注射（ＩＣＩ），并选择罂粟碱／酚妥拉明进行对照，结果表明，硝普钠ＩＣＩ后：（１）阴茎外形性状（长度、周径等）明显改变。（２）Ｖｉｒａｇ硬度计点表明硝普钠与罂粟碱／酚妥拉明效果之间无明显差别。（３）所有测试患者无一例出现低血压或局部不适等副反应，与罂粟碱／酚妥拉明相比各有优劣，但总体差异不大，这充分表明，硝普钠作为一种ＮＯ供体可导致阴茎平滑肌松弛，血窦充盈阴茎勃起，其副反应较小，有其临床应用之价值。     

糖皮质激素对肿瘤坏死因子引起的大鼠中性粒细胞粘附的影响

采用中性粒细胞（ＰＭＮ）与玻璃珠粘附和ＰＭＮ与血管内皮细胞（ＥＣ）粘附两种模型，以肿瘤坏死因子（ＴＮＦ），作为ＰＭＮ的刺激因子，研究糖皮质激素（ＧＣ）对ＴＮＦ引起的大鼠ＰＭＮ粘附的影响，同时给予糖皮质激素受体（ＧＲ）阻断剂ＲＵ３８４８６观察ＧＲ在粘附中的作用。结果发现，ＴＮＦ能明显增强大鼠ＰＭＮ的粘附（Ｐ＜０．０１）；Ｄｅｘ不能抑制经ＴＮＦ预处理的ＰＭＮ的粘附（Ｐ＞０．０５），但有一定的预防作用；经ＴＮＦ预处理再同时给予Ｄｅｘ和ＲＵ３８４８６的ＰＭＮ粘附同样明显增强（Ｐ＜０．０１）。     

Kelfiprim,a new sulpha-trimethoprim combination,versus cotrimoxazole,in the treatment of urinary tract infections: A multicentre,double-blind trial

Summary A new combination of trimethoprim with a sulphonamide, named Kelfiprim, differs from cotrimoxazole in that: a) the sulpha drug is sulphamethopyrazine instead of sulphamethoxazole; b) the trimethoprim to sulpha ratio is 5:4 instead of 1:5;c) the presence of a long-acting sulphonamide allows the administration of a daily dose of one capsule, following an initial loading dose of two capsules; d) a reduced amount of trimethoprim is given, as compared to cotrimoxazole, without any decrease of efficacy. Kelfiprim [KP] was compared to contrimoxazole [Co] in a multicentre double blind trial. Sixty four patients suffering from acute and chronic infections of the upper and lower urinary tract entered the study. Urine sterilisation and clinical improvement without relapses showed no differences from the two treatment groups. Tolerance was excellent except in two patients, one treated with KP and the other treated with Co, who showed a transient exanthema.     

Time-dependent hexaminolaevulinate induced protoporphyrin IX distribution after topical application in patients with cervical intraepithelial neoplasia: A fluorescence microscopy study

BACKGROUND AND OBJECTIVES: Compared to the conventional management of cervical intraepithelial neoplasia (CIN) the potential advantage of photodynamic therapy (PDT) for the treatment of cervical human papilloma virus (HPV)-related disease encompasses a minimal invasive procedure with reduced risk of profuse bleeding as a consequence of conization, and possibly more favorable long-term results avoiding cervical stenosis. At present little is known about the precise time-dependent distribution and histological localization of hexaminolaevulinate (HAL) induced protoporphyrin IX (PPIX) fluorescence in healthy tissue and in CIN. The aim of this study was to use ex vivo fluorescence microscopy to determine whether PPIX is selectively induced by neoplastic cells of the cervical epithelium at various times after topical application. STUDY DESIGN/MATERIALS AND METHODS: Cold cream containing 0.5% HAL was applied by means of cervical cap over various periods of time. We analyzed 52 healthy cervical mucosa and 84 CINs. RESULTS: At time delay 100 (+/-10) minutes, high epithelial fluorescence and a significant selectivity between epithelium and underlying lamina propria was found. By contrast, no significant difference between healthy and neoplastic tissues, or between low and high-grade epithelial dysplasia (P > or = 0.05), was observed at any time point. CONCLUSIONS: Application of HAL 0.5% cream to the cervix induced selective fluorescence in epithelial cells. The optimal ratio with a homogeneous PPIX distribution was obtained after 100 ( +/- 10) minutes cream application, which should be evaluated further for PDT.     

An updated review on the principles of intraoperative neurophysiological monitoring and the anaesthetic considerations

It is known that any surgery to the nervous system poses risks to neural structures and their surrounding structures. These mechanisms of injury are the result of mechanical manipulations, haemodynamic alterations, chemical or thermal injuries. Intraoperative neurophysiological monitoring (IONM), using various modalities, is employed to facilitate the assessment of the functional integrity of neural structures, and it is used to provide a real-time alerting system when changes caused by surgically induced insults are detected. The primary goal of IONM is reducing the risk of postoperative neurological deficits during these surgical procedures. It is used to provide information that allows the surgeon to correct any surgical interventions that may have compromised these systems and this also in turn provides guidance on what neurological deficits to anticipate postoperatively. Apart from being utilized as an alerting system to avoid catastrophic outcomes, IONM also assists as a guidance system using stimulation techniques to map out eloquent areas within the cortex, allowing identification of specific neuronal structures, particularly when landmarks cannot be easily recognized due to infiltration by pathological lesions.In this article, we focus on updating our previous paper published in 2019 and again, to provide attention to the various neurophysiological modalities that are employed in IONM. We will look at the basic underlying physiological principles and their individual indications for use clinically. We will explain the information that each modality provides. Importantly, and the primary reason for this article, we look at the various anaesthetic agents, their effects on each neurophysiological modality and other anaesthetic considerations such as haemodynamic and temperature effects. We will also recommend the use of an alert checklist for the multidisciplinary team should an intraoperative alert be issued during surgical procedures.