加味旋覆代赭汤对非酸反流所致慢性食管病变大鼠肠化及 CDX2、DLK1表达的影响

目的:采用十二指肠-食管吻合术(胃全切)建立大鼠十二指肠-食管非酸反流模型,观察加味旋覆代赭汤对模型食管肠化及CDX2、DLK1表达的影响。方法:成年雄性Wistar大鼠80只,随机分为假手术组、模型组、加味旋覆代赭汤组及铝碳酸镁片(达喜)组,每组20只。除假手术组外,其余三组均采用十二指肠-食管吻合术(胃全切)建立大鼠十二指肠-食管非酸反流模型,建模25周时,加味旋覆代赭汤组经口灌服加味旋覆代赭汤(相当于30 g生药/kg),每日1次,连续3周;达喜组经口灌服达喜药液(300 mg/kg),每日1次,连续3周;模型组给予同体积蒸馏水经口灌服,每日1次,连续3周。治疗3周后,连同假手术组,共4组动物,麻醉状态下打开胸腹腔,剪取食管下段及吻合段,纵行剖开,进行大体观察,对食管下端紧邻吻合口处组织行HE染色,行食管病理组织学检查并评分,另采用免疫组化法进行CDX2及Notch信号分子-DLK1染色,观察其表达情况。结果:大体观察发现模型组食管下段显著增粗,纵行剖开食管,可见下端食管显著增厚,食管下段黏膜呈广泛显著不规则隆起,外观颜色呈白斑样改变;加味旋覆代赭汤组食管大体病理较模型组显著减轻,达喜组食管大体改变较模型组无显著差别。模型组轻、中、重度食管炎发生率分别为2/15(13.33%)、3/15(20.00%)、10/15(66.67%);加味旋覆代赭汤组分别为11/17(64.70%)、3/17(17.65%)、3/17(17.65%),与模型组比较炎症程度显著降低(P<0.01),达喜组分别为3/16(18.75%)、4/16(25.00%)、9/16(56.25%),与模型组差异无显著性(P>0.05)。加味旋覆代赭汤组的Barrett食管发生率为2/17(11.8%),显著低于模型组的7/15(46.7%),差异有统计学意义(P<0.05),达喜组为7/16(43.8%),与模型组差异无统计学意义(P>0.05)。加味旋覆代赭汤组的CDX2、DLK1表达水平均显著低于模型组(P<0.01),而达喜组与模型组比较差异无统计学意义(P>0.05)。结论:加味旋覆代赭汤可通过抑制CDX2及Notch信号分子DLK1表达来抑制肠化,减少非酸反流所致Barrett食管的发生。     

大鼠椎间盘软骨细胞中Bax、Bcl-2及 Caspase-8的表达

目的:探讨椎间盘软骨细胞在正常与凋亡状态下,Bax、Bcl-2及Caspase-8的表达变化情况。方法:使用酶消化法获取雄性SD大鼠椎间盘软骨细胞,抗Fas抗体诱导凋亡,免疫组化法(SP)检测大鼠正常与凋亡椎间盘软骨细胞的Bax、Bcl-2及Caspase-8的表达,CMIAS-99B型医学图像系统进行表达强度的分析,进而比较分析大鼠正常与凋亡椎间盘软骨细胞的Bax、Bcl-2及Caspase-8的表达变化情况。结果:正常组的Bax和Caspase-8轻度表达,凋亡模型组的Bax和Caspase-8表达明显;而正常组的Bcl-2高度表达,凋亡模型组的Bcl-2轻度表达,正常和凋亡各组均有非常显著差异(P<0.01)。结论:凋亡的大鼠椎间盘软骨细胞内Bax、Bcl-2、Caspase-8的表达与正常状态相比有明显变化。     

活血消异方改善子宫内膜异位症大鼠卵泡发育的机制探讨

目的 从卵巢颗粒细胞凋亡与自噬相互作用的角度，探讨活血消异方对子宫内膜异位症大鼠卵泡发育的影响及作用机制。方法 将动情周期正常的48只SD大鼠分为空白组、假手术组各10只，剩余的分为供体组(n=5)和受体组(n=23)用于造模，造模成功后的20只大鼠又分为模型组、活血消异方组各10只。活血消异方组予活血消异方汤剂2 ml灌胃，空白组、假手术组、模型组均予蒸馏水2 ml灌胃，各组连续灌胃15 d后，检测大鼠血清活性氧(ROS)、总超氧化物歧化酶(T-SOD)、过氧化氢酶(CAT)水平。取各组大鼠的卵巢组织行HE染色，光镜下观察各级卵泡形态并计数；免疫组化染色定位颗粒细胞，并比较凋亡相关因子Bax、Bcl-2、Caspase-3的表达；TUNEL染色观察颗粒细胞凋亡情况并计算凋亡率；蛋白免疫印迹法(Western blot)检测卵巢组织中凋亡及自噬相关蛋白p-JNK、Bax、Bcl-2、Caspase-3、Beclin-1、LC3Ⅱ的表达；电镜观察各组大鼠卵巢颗粒细胞自噬体形态，并计算自噬体胞质面积比。结果 (1)与空白组、假手术组比较，模型组血清ROS含量显著升高，T-SOD、CAT水平...     

黄芪甲苷预处理对大鼠肠缺血再灌注损伤的影响

目的:探讨黄芪甲苷(AS-Ⅳ)预处理对大鼠肠缺血再灌注损伤的影响。方法:30只成年雄性Wistar大鼠随机分为假手术(Sham)组、模型(I/R)组、AS-Ⅳ预处理(AS-Ⅳ)组。I/R组和AS-Ⅳ组采用无创微动脉夹夹闭肠系膜上动脉60 min再灌注120 min的方法建立大鼠肠缺血再灌注损伤模型。AS-Ⅳ组于造模前60 min时采用60 mg/kg AS-Ⅳ对大鼠进行灌胃预处理。于再灌注120 min时取小肠组织,采用HE染色法测定小肠组织病理学变化,ELISA法测定大鼠血清中TNF-α、HMGB-1、SOD和CAT的表达情况,荧光定量PCR法测定小肠组织中Bcl-2、Bax和Caspase-9的mRNA情况,Western blotting法测定小肠组织中紧密连接蛋白occludin和ZO-1的表达情况。结果:与Sham组比较,I/R组小肠组织病理学Chiu评分升高,TNF-α和HMGB-1的表达水平升高,SOD和CAT的表达水平降低,Bcl-2、Bax和Caspase-9的mRNA水平升高,occludin和ZO-1的蛋白表达水平降低(P0.05);与I/R组比较,AS-Ⅳ组I/R组小肠组织病理学Chiu评分降低,TNF-α和HMGB-1的表达水平降低,SOD和CAT的表达水平升高,Bcl-2、Bax和Caspase-9的mRNA水平降低,occludin和ZO-1的蛋白表达水平升高(P0.05)。结论:AS-Ⅳ灌胃预处理能够显著改善大鼠肠I/R损伤,其机制可能与减轻机体炎症反应和氧化应激水平、抑制肠道细胞的凋亡发生以及调节紧密连接蛋白表达有关。     

猪苓汤对阿霉素肾病大鼠肾脏AQP2表达的影响

目的:探讨猪苓汤对阿霉素肾病大鼠水通道蛋白2表达的影响。方法:110只SPF级雄性SD大鼠随机分为空白组、假手术组、模型组,尾静脉注射阿霉素14 d后,24 h尿蛋白定量≥100 mg提示模型复制成功,将模型大鼠再次随机分为模型组、猪苓汤组、呋塞米组,分别于灌胃2、4、6、8周处死部分大鼠,留取血液、尿液、肾脏标本,半自动生化仪检测尿蛋白含量、肾功能等生化指标,HE染色观察肾脏病理改变,RT-PCR法检测AQP2 mRNA表达,Western Blot法检测AQP2蛋白表达。结果:(1)生化指标:灌胃8周,猪苓汤组平均尿量高于模型组(P<0.05),其他生化指标比较,各造模组之间差异无统计学意义;(2)AQP2 mRNA:注射后14 d,模型组平均表达量高于空白组,差异具有统计学意义(P<0.05),猪苓汤组在灌胃2、4、6、8周时AQP2 mRNA平均表达量均低于模型组(P<0.05);(3)AQP2蛋白:注射后14 d,模型组平均表达量高于空白组,差异具有统计学意义(P<0.05),猪苓汤组在灌胃4、6、8周时AQP2蛋白平均表达量均低于模型组(P<0.05)。结论:猪苓汤的利尿作用可能通过下调AQP2 mRNA和蛋白表达而实现。     

人参皂苷Rg1对梗阻性肾病大鼠肾脏Klotho表达及肾小管上皮细胞凋亡的影响

目的:观察人参皂苷Rg1(ginsenoside rg1,G-Rg1)对单侧输尿管梗阻(unilateral ureteral obstruction,UUO)大鼠肾脏Klotho表达及肾小管上皮细胞(renal tubular epithelial cell,RTC)凋亡的影响。方法:24只清洁级SD大鼠随机分为三组:对照组、模型组、治疗组;模型组大鼠采用UUO的方法建立肾间质纤维化(renal interstitial fibrosis,RIF)模型,治疗组在UUO的基础上,给予G-Rg1腹腔注射,其余大鼠给予腹腔注射等剂量生理盐水。2周后检测各组大鼠的肾功能,HE及Masson染色观察肾脏病理改变;TUNEL检测RTC凋亡情况,RT-PCR及Western blot检测肾组织Klotho表达变化,Western Blot检测凋亡相关蛋白Bcl-2、Bax的表达情况。结果:与对照组比较,模型组大鼠血清肌酐、尿素氮明显上升,肾脏病理损害加重,RTC凋亡增加;同时模型组肾组织Klotho mRNA及蛋白的表达均减少,伴随凋亡蛋白Bax表达增多及凋亡抑制蛋白Bcl-2表达减少;与模型组比较,治疗组大鼠肾功能好转,肾脏病理损害缓解,RTC凋亡减轻;同时肾组织Klotho mRNA及蛋白表达水平升高,伴随凋亡蛋白Bax表达减少及凋亡抑制蛋白Bcl-2表达增多。结论:G-Rg1可以上调UUO大鼠肾脏Klotho的表达,进而调节凋亡相关蛋白Bax及Bcl-2的表达水平,从而发挥抑制UUO模型中RTC凋亡的作用。     

L-精氨酸对局灶性脑缺血大鼠神经元凋亡的影响

目的 评价L-精氨酸(L-arg)对局灶性脑缺血大鼠神经元凋亡的影响.方法 健康雄性SD大鼠56只,体重250～300 g,随机分为7组(n=8):假手术组(SH组)、脑缺血2 h组(Is1组)、脑缺血2 h L-arg治疗组(L-arg1组)、脑缺血6 h组(IS2组)、脑缺血6 h L-arg治疗组(L-arg2组)、脑缺血12h组(IS3组)及脑缺血12 h L-arg治疗组(L-arg3组).采用线栓法制备大鼠大脑中动脉阻塞模型.各L-arg治疗组分别于脑缺血后腹腔注射L-精氨酸500 mg/kg,2次/d,治疗3 d;IS组给予等容量生理盐水.治疗3 d后取脑,测定缺血区域神经元凋亡率、Caspase-3蛋白、Bcl-2蛋白和Bax蛋白的表达水平.结果 与SH组比较,IS1组、IS2组和IS3组神经元凋亡率升高,Caspase-3蛋白和Bax蛋白表达上调,Bcl-2/Bax蛋白比值降低(P＜0.01);与IS1组和IS2组比较,L-arg1组和L-arg2组神经元凋亡率降低,Caspase-3蛋白和Bax蛋白表达下调,Bcl-2蛋白表达上调,Bcl-2/Bax蛋白比值升高(P＜0.01).IS3组与L-arg3组上述指标差异无统计学意义(P＞0.05).结论 L-arg可减少脑缺血早期大鼠神经元凋亡,具有一定的治疗作用,其机制可能与下调Caspase-3蛋白表达、调节Bcl-2/Bax蛋白平衡有关.     

上调NICD对人牙周膜干细胞增殖及迁移能力的影响

目的:研究上调Notch胞内区域(Notch Intracellular Domain,NICD)对人牙周膜干细胞增殖及迁移能力的影响。方法:将细胞分为上调组、对照组及空白组,使用MTT比色法检测细胞增殖能力,流式细胞仪检测细胞凋亡能力,采用细胞划痕实验检测细胞迁移情况,并使用Western blot检测人牙周膜干细胞中Wnt/β-catenin信号通路蛋白(TGF-β、BMP-2、β-caten、LEF-1、Cyclin D1、Caspase-3、Bax、Bcl-2)表达量。结果:对照组各时间点的人牙周膜干细胞增殖率、碱性磷酸酶活性、迁移、黏附个数、TGF-β、BMP-2、β-caten、LEF-1、Cyclin D1、Bcl-2蛋白表达量均低于空白组,且凋亡率、Caspase-3、Bax蛋白表达量高于空白组,差异具有统计学意义(P<0.05)。上调组各时间点人牙周膜干细胞增殖率、碱性磷酸酶活性、迁移、黏附个数、TGF-β、BMP-2、β-caten、LEF-1、Cyclin D1、Bcl-2蛋白表达量均高于空白组和对照组,且凋亡率、Caspase-3、Bax蛋白表达量均低于空...     

Ang-1在去卵巢骨质疏松症大鼠骨组织中表达变化以及中医不同治法对其影响的实验研究

目的观察去卵巢骨质疏松症模型大鼠的骨组织中血管生成素1(Ang-1)mRNA和蛋白含量变化,及中药不同方剂对其影响,探讨中药组方治疗骨质疏松的可行性。方法骨质疏松症模型的建立采用切除雌性大鼠双侧卵巢的方法。运用补肾填精中药复方、活血化瘀中药复方、补肾活血中药复方对模型大鼠灌胃12周,用骨疏康作为阳性药物对照组,还有正常组和模型空白组。RT-PCR检测各组骨组织Ang-1mRNA相对表达量;ELISA法检测各组骨组织Ang-1蛋白含量。结果去卵巢骨质疏松症模型大鼠的骨组织中Ang-1mRNA相对表达量与正常组比较,显著降低(P0.01);与模型空白组比较,各个用药组Ang-1mRNA相对表达量显著升高(P0.01);但各个用药组间比较无统计学意义。去卵巢骨质疏松症模型大鼠骨组织中Ang-1蛋白含量与正常组比较,显著降低(P0.01);与模型空白组比较,各个用药组Ang-1含量显著升高(P0.01);各个用药组间比较,补肾活血组Ang-1含量升高最为显著,与补肾填精组和活血化瘀组比较有显著性差异(P0.01)。结论补肾活血中药复方可提高Ang-1mRNA相对表达量及其蛋白含量,且优于单纯的补肾填精和活血化瘀中药复方,起到防止骨质疏松症的作用。     

补肾健脾活血方调控Apaf-1/CHOP-Caspase-9/Bcl-2通路抑制成骨细胞氧化应激

目的 探讨补肾健脾活血方对减轻大鼠成骨细胞氧化应激损伤和调控线粒体介导的细胞凋亡的机制。方法 提取大鼠原代成骨细胞,通过CCK8实验检测补肾健脾活血方干预下对成骨细胞增殖活性的影响、碱性磷酸酶(alkaline phosphatase, ALP)染色检测成骨细胞ALP活性、茜素红染色检测成骨细胞的矿化情况、实时定量PCR检测Apaf-1和CHOP基因的mRNA表达、蛋白质印迹检测Caspase-9和Bcl-2蛋白表达量。结果 CKK8实验和碱性磷酸酶染色表明氧化应激损伤的大鼠成骨细胞增殖活性显著降低,成骨分化受抑制,同时Apaf-1和CHOP的mRNA表达以及Caspase-9的蛋白表达明显增加(P≤0.01),Bcl-2蛋白表达则显著减少(P≤0.01);经补肾健脾活血方干预后,成骨细胞增殖活性得到明显的增强、成骨分化增加,Apaf-1和CHOP的mRNA及Caspase-9的蛋白表达均显著减少(P<0.05),而Bcl-2的蛋白表达则显著升高(P≤0.000 1)。结论 补肾健脾活血方可以促进成骨细胞增殖和成骨分化,显著降低Apaf-1和CHOP的mRNA及Caspase-9...     

The TEG® vs the ROTEM® thromboelastography/thromboelastometry systems

We have evaluated the TEG^® thromboelastograph and the ROTEM^® thromboelastometer, two point-of-care devices that measure blood coagulation. During a one-week period, seven consultant anaesthetists, one consultant haematologist, one associate specialist anaesthetist and two senior trainee anaesthetists were trained by the manufacturers and set up, calibrated and used both systems, after which their views were obtained and specific technical/support information was sought from the manufacturers using a questionnaire. Although the devices shared common features, they differed in complexity and aspects of ease of use, and in their purchase and running costs.     

Laser Resurfacing of the Neck with the Combined CO2/Er:YAG Laser

BACKGROUND: Several clinical studies on laser resurfacing on the neck have yielded variable results with adverse sequelae of hypopigmentation and scarring using the CO2 laser. The Er:YAG laser on the neck resulted in an improved appearance with no adverse sequelae. The combined CO2/Er:YAG laser is a laser that combines a low fluence CO2 laser with the Er:YAG laser in a near simultaneous beam. OBJECTIVE: To study the effects of the CO2/Er:YAG laser on the neck. The decreased nonspecific thermal damage of the CO2/Er:YAG laser should result in decreased postoperative erythema and improved wound healing. METHODS: Eleven patients were treated with the CO2/Er:YAG laser on the neck as well as the face at identical settings. The face was treated with four passes while the neck was treated with two passes. The patients were evaluated for the first 2 weeks then 3-6 months postoperatively. Patients were asked about treatment satisfaction and improvement in skin texture and color using a 25% scale. Skin texture and color, as well as improvement in wrinkling and adverse results were judged by a dermatologist using a 0-4 scale. RESULTS: Moderate improvement was seen in skin color while a higher degree of improvement was seen in skin texture and wrinkling in all 11 patients. No adverse sequelae, including hypopigmentation or scarring, were observed. The majority of patients had a 75-100% improvement in skin texture and color in addition to a 75-100% overall satisfaction rating. CONCLUSIONS: There is a higher degree of overall patient satisfaction, as well as improvement in skin texture and skin color, compared to patients treated with the Er:YAG laser alone. The CO2/Er:YAG laser is a highly effective laser for neck resurfacing with no adverse sequelae to date.     

Icariin improves the sexual function of male mice through the PI3K/AKT/eNOS/NO signalling pathway

We aimed to investigate the effect and mechanism of icariin on male sexual function. Forty‐eight Crl:CD1(ICR) male mice were randomly divided into control, low‐, medium‐ and high‐dose icariin group (intragastric administration of 50, 100 and 200 mg/kg/d for 21 days). Mating experiment was then performed at a ratio of 1: 3 (male: female). The mating behaviours of male mice were recorded. The genital indexes and serum testosterone, nitric oxide (NO), hypothalamic dopamine (DA) and 5‐ hydroxy tryptophan (5‐HT) concentrations were measured. The expression of endothelial nitric oxide (eNOS), phosphatidylinositol tallow alcohol 3‐kinase (PI3K) and phosphorylated protein kinase (p‐AKT) in penile tissue was detected by Western blot. All icariin groups exhibited shorter capture latency and ejaculation latency, increased number of capture and ejaculation, higher capture and ejaculation rate, and higher testicular and prostate indexes compared with controls (p < .001). These groups had higher serum testosterone and NO concentrations (p < .001), hypothalamic DA and 5‐HT levels, and eNOS, PI3K and phosphorylated AKT expressions in penile tissue (p < .05). The effect of icariin was dose‐dependently increased. Our study suggests that icariin improves the sexual function of male mice, which might be associated with the hypothalamic–pituitary–gonadal axis and the PI3K/AKT/eNOS/NO signalling pathway.     

Clinical relevance of the B12/N2O interaction A report of a seminar

The interaction of nitrous oxide and vitamin B₁₂ and its implications are not the exclusive territory of any one discipline. The initial discovery was by a chemist but it is of obvious relevance to anaesthetists and intensivists; some complications are neurological others haematological. The interaction provides an extremely important research tool as the first easily available B₁₂-deficient animal model. Finally there are implications for exposure to contaminated atmospheres in hospitals and in industry.