右美托咪定对子宫内膜癌细胞增殖、凋亡及Wnt/β-catenin通路的影响

目的 探究右美托咪定对子宫内膜癌细胞增殖、凋亡及Wnt/β-连环蛋白（β-catenin）通路的影响。方法 将Ishikawa、RL95-2细胞分为对照组,右美托咪定1、10、100 nmol/L组,右美托咪定（100 nmol/L）＋LiCl组。CCK-8法和EdU检测细胞增殖,流式细胞术检测细胞凋亡,Western blotting检测增殖细胞核抗原（PCNA）、B淋巴细胞瘤-2蛋白（Bcl-2）、Bcl-2相关X蛋白（Bax）、β-连环蛋白（β-catenin）、c-Myc基因（c-Myc）、细胞周期蛋白D1（Cyclin D1）表达水平。构建子宫内膜癌裸鼠模型,分为对照组、右美托咪定组、右美托咪定＋LiCl组,测量肿瘤质量与体积,苏木精–伊红（HE）染色观察移植瘤组织形态,免疫组化法检测移植瘤组织Ki-67、β-catenin蛋白表达。结果 与对照组相比,不同浓度右美托咪定处理后Ishikawa、RL95-2细胞吸光度（A）值、EdU阳性细胞率、迁移细胞数、侵袭细胞数、PCNA、Bcl-2、β-catenin、c-Myc、Cyclin D1蛋白表达显著降低,凋亡率、Bax表达上升（P＜0.05）;与右美托咪定100 nmol/L组相比,右美托咪定＋LiCl组A值、EdU阳性细胞率、迁移细胞数、侵袭细胞数、PCNA、Bcl-2、β-catenin、c-Myc、Cyclin D1蛋白表达显著升高,凋亡率、Bax表达下降（P＜0.05）。右美托咪定能抑制移植瘤质量和体积,促进肿瘤凋亡,降低β-catenin、Ki-67蛋白表达（P＜0.05）;LiCl能逆转右美托咪定对移植瘤的抑制作用（P＜0.05）。结论 右美托咪定能抑制子宫内膜癌细胞增殖,促进凋亡,其作用机制可能与抑制Wnt/β-catenin信号通路有关。     

益智康脑丸对老年性痴呆大鼠行为学及脑组织Bcl-2和Bax蛋白表达的影响

目的:探讨益智康脑丸对老年性痴呆(AD)大鼠行为学及脑组织凋亡蛋白的影响。方法:复制D-半乳糖AD大鼠模型。实验分为4组,即空白、模型、益智康脑丸和脑复康组,检测大鼠行为学及脑组织抑制细胞凋亡基因Bcl-2和促凋亡基因Bax表达指标的改变。结果:模型组与空白组比较,学习记忆能力显著下降,Bcl-2/Bax比值下降;治疗组与模型组比较,学习记忆能力显著改善,Bcl-2/Bax比值增加。结论:益智康脑丸具有较好的治疗AD作用,其机制可能与促进AD大鼠脑组织Bcl-2的表达、降低Bax的表达有关。     

PI3K-Akt-HIF-α信号通路在右美托咪定减轻大鼠肝缺血/再灌注致肺损伤中的作用

目的:通过研究右美托咪定对大鼠肝缺血/再灌注致急性肺损伤中的作用及其可能作用机制.方法:建立大鼠肝缺血/再灌注肺损伤模型,以随机数字表法分为假手术组、肝缺血/再灌注组(I/R组)、I/R+右美托咪定组(D组)和I/R+右美托咪定组+wortmannin(DW组),通过ELISA和Western blot检测细胞炎症因子及PI3K-Akt-HIF-α信号传导通路蛋白的表达.结果:I/R组中MDA和MPO含量、 细胞炎症因子表达及p-Akt和HIF-α蛋白含量较假手术组均明显升高;D组中上述肺损伤指标较I/R组均显著下调;与D组相比,DW组中MDA和MPO含量、细胞炎症因子表达及p-Akt和HIF-α蛋白含量均上调.     

PI3K/Akt信号传导通路在右美托咪定抗大鼠肢端缺血/再灌注致急性肺损伤中的作用

目的:探讨PI3K/Akt信号传导通路在右美托咪定对大鼠肢体缺血/再灌注致急性肺损伤中的作用.方法:通过止血带捆扎大鼠双后肢根部从而建立大鼠缺血/再灌注肺损伤模型,通过随机数字表法分为对照组、缺血/再灌注组(I/R组)、I/R+右美托咪定组(D组)和I/R+右美托咪定组+LY294002(DL组),通过ELISA和Western blot检测细胞炎症因子及PI3K/Akt信号传导通路蛋白的表达.结果:I/R组中MDA和MPO含量、细胞炎症因子表达及p-Akt蛋白含量较对照组均明显升高;D组中上述肺损伤指标较I/R组均显著下调;与D组相比,DL组中MDA和MPO含量、细胞炎症因子表达及p-Akt蛋白含量均上调.     

右美托咪定对全脑缺氧缺血损伤大鼠海马内星形胶质细胞VEGF表达的影响

目的探讨右美托咪定对全脑缺氧缺血损伤大鼠海马内星形胶质细胞中血管内皮细胞生长因子(Vascular endothelial growth factor,VEGF)表达的影响。方法取SD大鼠24只,体重250~280 g,采用随机数字表法将其分为3组(n=8):假手术组(S组)、常规复苏组(C组)和右美托咪定组(D组)。C组和D组采用窒息后心跳骤停法建立大鼠全脑缺氧缺血损伤模型,S组大鼠仅行气管插管不夹闭窒息,D组于气管夹闭前尾静脉注射负荷剂量右美托咪定4μg/kg,S组和C组分别给予等容量生理盐水。各组大鼠于自主循环恢复后12、24、48及72 h进行神经功能缺失评分(Neuropathy disability score,NDS),于72 h测定NDS后处死大鼠,取海马组织,采用TUNEL法标记检测海马神经元凋亡情况,采用免疫荧光双标法测定各组大鼠海马内星形胶质细胞VEGF的表达情况。结果与S组比较,C组各时点NDS评分均显著升高(P<0.05);D组大鼠在12 h时NDS评分与C组比较差异无统计学意义(P>0.05),D组大鼠在24、48和72 h三个时点NDS评分显著低于C组(P<0.05)。与S组比较,C组大鼠海马内神经元凋亡率和星形胶质细胞VEGF的表达均升高;与C组比较,D组大鼠海马内神经元凋亡率降低,星形胶质细胞VEGF的表达升高(P<0.05)。结论右美托咪定可减轻全脑缺氧缺血损伤大鼠脑损伤,降低海马内神经元的凋亡率,其机制可能与促进海马内星形胶质细胞上VEGF的表达有关。     

阿魏酸钠对脓毒症大鼠肾组织Bcl-2和Bax表达的影响

目的观察阿魏酸钠(活血化瘀中成药)对脓毒症大鼠肾脏细胞凋亡及Bcl-2、Bax的影响。方法用盲肠穿孔结扎术制作脓毒症模型;24只SD大鼠随机分为模型组、实验组、假手术组和正常组(均n=6);术后6 h处死大鼠;用光镜观察肾脏病理改变,用免疫组化法检测Bcl-2及Bax的表达。结果模型组Bax表达明显升高(P<0.01),见细胞凋亡;实验组Bax表达明显降低而Bcl-2表达明显升高(P<0.01),病理损伤有改善。结论阿魏酸钠通过下调Bax、上调Bcl-2的表达,提高Bcl-2/Bax比值,抑制肾小管上皮细胞凋亡。     

右美托咪定对CCI大鼠脊髓GFAP表达的影响

目的 观察右美托咪定对神经病理性疼痛大鼠脊髓GFAP蛋白表达的影响;方法 雄性SD大鼠32只,随机分为4组:假手术组(Sham组)、模型组(Model组)、右美托咪定50μg·kg-1(D1组)和右美托咪定100μg·kg-1(D2组),各组于手术后即刻至术后7天,每天腹腔注射1次药物,假手术组和模型组给予等体积的生理盐水.于7d,造模前和造模后1、3、5、7d观察各组大鼠的机械痛阈(PWPT)变化,于造模后7d免疫荧光组织化学法观测各组脊髓胶质纤维酸性蛋白(Glial Fibrillary Acidic Protein,GFAP)表达;结果 大鼠建模1天后,PWPT显示CCI大鼠的机械痛阈均出现明显下降,右美托咪定干预的D1组在7d对CCI大鼠的痛阈有所改善(P<0.05),D2组对大鼠的痛阈有明显改善,在3d及以后提高明显(P<0.01),同时,7d时模型组GFAP的表达明显增强,D1组较模型组表达减弱(P<0.05),D2组较模型组差异有显著的统计学意义(P<0.01).结论 右美托咪定对神经病理性疼痛的抑制作用可能与抑制脊髓GFAP的表达有关.     

依达拉奉右莰醇通过铁死亡-脂质过氧化通路对脑出血大鼠神经保护的作用机制

目的 探讨依达拉奉右莰醇对脑出血大鼠的神经保护作用及血肿周围脑组织脂质过氧化的影响。方法将128只SD大鼠随机分为假手术组、脑出血组、依达拉奉组和依达拉奉右莰醇组，每组32只。除假手术组外，其余组大鼠构建急性脑出血模型，依达拉奉组、依达拉奉右莰醇组于造模后分别腹腔注射依达拉奉6 mg/kg、依达拉奉右莰醇7.5 mg/kg，每12 h注射1次，假手术组和脑出血组腹腔注射等量生理盐水。术后1 d、3 d、7 d和14 d按Garcia评分标准进行神经功能评分，HE染色观察血肿周围脑组织病理变化，化学荧光法检测血肿周围脑组织活性氧（ROS）含量，微量酶标法检测血肿周围脑组织还原型谷胱甘肽（GSH）含量，蛋白免疫印迹法检测血肿周围脑组织谷胱甘肽过氧化物酶4(GPX4)、长链脂酰辅酶A合成酶4(ACSL4)和磷脂胆碱酰基转移酶3(LPCAT3)表达。结果 与假手术组比较，脑出血组大鼠神经功能评分降低，血肿周围脑组织出现大量炎性细胞浸润及神经细胞变性，ROS含量、ACSL4和LPCAT3蛋白表达水平升高，GSH含量、GPX4蛋白表达水平降低（P<0.05）；与脑出血组比较，依达拉奉组和依达...     

依达拉奉对颅脑损伤大鼠脑组织Bcl-2和Bax表达的影响

目的:观察依达拉奉对大鼠颅脑损伤后脑组织中Bcl-2和Bax表达的影响和对脑组织的保护作用。方法:将30只SD大鼠随机分为假手术组、模型组和依达拉奉治疗组。采用自由落体法建立大鼠颅脑损伤模型,免疫组织化学法和RT-PCR法观察脑组织Bcl-2和Bax的表达。结果:大鼠颅脑损伤后,模型组和治疗组Bcl-2和Bax的表达较假手术组显著增多(P<0.05);予依达拉奉治疗后,治疗组Bcl-2的表达较模型组显著上调(P<0.05),Bax的表达较模型组显著下调(P<0.05)。结论:依达拉奉可上调Bcl-2的表达,抑制Bax的表达,减少颅脑损伤后神经细胞凋亡,这可能是其发挥脑保护作用的机制之一。     

血糖水平对大鼠脑挫裂伤后伤灶周围神经细胞中Bcl-2和Bax mRNA表达水平的影响

目的 研究大鼠脑挫裂伤后不同血糖水平对伤灶周围神经细胞中凋亡相关基因Bcl- 2和Bax mRNA表达水平的影响.方法 将60只健康雄性SD大鼠随机分成对照组和实验组,以自由落体法制造大鼠脑挫裂伤模型后,颈静脉置管输入50%葡萄糖溶液建立高血糖动物模型,对照组以相同方法输入等量生理盐水溶液.在伤后即刻、6、12、24、48h 5个时间点,抽提大鼠伤灶周围组织总RNA,RT-PCR方法分析两组大鼠伤灶周围的脑组织中Bcl-2和Bax mRNA表达水平.结果 正常脑组织中Bcl-2 mRNA表达几乎为阴性;脑挫裂伤后早期伤灶周围神经细胞Bcl-2 mRNA表达明显增加,在伤后24h达到高峰,之后渐下降.而正常脑组织中有Bax mRNA表达,脑挫裂伤后早期伤灶周围脑细胞Bax mRNA表达亦有明显增加,伤后12h达到高峰,之后渐下降.伤后各时间点实验组大鼠Bcl-2 mRNA的表达水平较对照组大鼠明显减低,而Bax mRNA表达则明显升高.结论 脑挫裂伤后高血糖水平对挫裂伤灶周围神经细胞中Bcl- 2基因的表达有抑制作用,而对Bax基因的表达则有促进作用.脑挫裂伤后高血糖水平会加重伤灶周围神经细胞的凋亡.     

Synthesis,Cytotoxicity and Antileishmanial Activity of Some N-(2-(indol-3-yl)ethyl)-7-chloroquinolin-4-amines

We report herein the condensation of 4,7-dichloroquinoline (1) with tryptamine (2) and D-tryptophan methyl ester (3) . Hydrolysis of the methyl ester adduct (5) yielded the free acid (6) . The compounds were evaluated in vitro for activity against four different species of Leishmania promastigote forms and for cytotoxic activity against Kb and Vero cells. Compound (5) showed good activity against the Leishmania species tested, while all three compounds displayed moderate activity in both Kb and Vero cells.     

Efficacy of gut lavage,hemodialysis, and hemoperfusion in the therapy of paraquat or diquat intoxication

Clinical and in vitro investigations were carried out to test the efficacy of gut lavage, hemodialysis, and hemoperfusion in the treatment of poisoning with paraquat or diquat. In a patient suffering from diquat intoxication 130 times more diquat was removed by gut lavage 30 h after ingestion than was removed by complete aspiration of the gastric contents.Determination of in vitro clearances for paraquat and diquat by hemodialysis showed that, at serum concentrations of 1–2 ppm, such as are frequently encountered in poisoning in man, toxicologically relevant quantities of herbicide cannot be removed from the body. At a concentration of 20 ppm, on the other hand, hemodialysis proved to be effective, the clearance being 70 ml/min at a blood flow rate of 100 ml/min. The efficacy of hemoperfusion with coated activated charcoal was on the whole better. Especially at concentrations around 1–2 ppm, the clearance values for hemoperfusion were some 5–7 times higher than those for hemodialysis.In a patient suffering from paraquat poisoning, both hemodialysis as well as hemoperfusion were carried out. The in vitro results could be confirmed: At serum concentrations of paraquat less than 1 ppm no clearance could be obtained by hemodialysis while by hemoperfusion with activated charcoal quite high clearance values were measured and the serum level dropped down to zero.

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Zusammenfassung Klinische Untersuchungen und Laboratoriumsversuche wurden durchgeführt, um die Wirksamkeit von Darmspülung, Hämodialyse und Hämoperfusion bei Paraquat- und Deiquat-Vergiftungen zu prüfen.Bei einem Patienten wurde 30 Std nach Deiquat-Aufnahme durch Darmspülung 130mal mehr Deiquat entfernt als durch vollständige Aspiration des Mageninhaltes. In vitro-Versuche ergaben, daß bei Blutserumkonzentrationen von 1–2 ppm, die bei Vergiftungen oft gemessen werden, durch Hämodialyse keine toxikologisch relevanten Paraquat- oder Deiquat-Mengen entfernt werden können. Dagegen erwies sich die Hämodialyse bei 20 ppm und einer Blutumlaufgeschwindigkeit von 100 ml/min mit einer Clearance von 70 ml/min als wirksam. Die Hämoperfusion mit beschicheter Aktivkohle war in diesen Versuchen aber eindeutig überlegen, denn insbesondere bei Konzentrationen um 1–2 ppm waren die Clearance-Werte 5–7mal höher als bei der Hämodialyse.Die in vitro-Ergebnisse wurden bei einem Patienten mit einer Paraquat-Vergiftung bestätigt: Bei Konzentrationen unter 1 ppm war die Hämodialyse wirkungslos, während durch Hämoperfusion relativ hohe Clearance-Werte erreicht wurden, so daß der Serumspiegel rasch unter die Nachweisgrenze abfiel.

     

In vitro studies on sterically stabilized liposomes (SL) as enzyme carriers in organophosphorus (OP) antagonism

This study describes a new approach for organophosphorous (OP) antidotal treatment by encapsulating an OP hydrolyzing enzyme, OPA anhydrolase (OPAA), within sterically stabilized liposomes. The recombinant OPAA enzyme was derived from Alteromonas strain JD6. It has broad substrate specificity to a wide range of OP compounds: DFP and the nerve agents, soman and sarin. Liposomes encapsulating OPAA (SL)* were made by mechanical dispersion method. Hydrolysis of DFP by (SL)* was measured by following an increase of fluoride ion concentration using a fluoride ion selective electrode. OPAA entrapped in the carrier liposomes rapidly hydrolyze DFP, with the rate of DFP hydrolysis directly proportional to the amount of (SL)* added to the solution. Liposomal carriers containing no enzyme did not hydrolyze DFP. The reaction was linear and the rate of hydrolysis was first order in the substrate. This enzyme carrier system serves as a biodegradable protective environment for the recombinant OP-metabolizing enzyme, OPAA, resulting in prolongation of enzymatic concentration in the body. These studies suggest that the protection of OP intoxication can be strikingly enhanced by adding OPAA encapsulated within (SL)* to pralidoxime and atropine.     

Lung disease and PKCs

The lung offers a rich opportunity for development of therapeutic strategies focused on isozymes of protein kinase C (PKCs). PKCs are important in many cellular responses in the lung, and existing therapies for pulmonary disorders are inadequate. The lung poses unique challenges as it interfaces with air and blood, contains a pulmonary and systemic circulation, and consists of many cell types. Key structures are bronchial and pulmonary vessels, branching airways, and distal air sacs defined by alveolar walls containing capillaries and interstitial space. The cellular composition of each vessel, airway, and alveolar wall is heterogeneous. Injurious environmental stimuli signal through PKCs and cause a variety of disorders. Edema formation and pulmonary hypertension (PHTN) result from derangements in endothelial, smooth muscle (SM), and/or adventitial fibroblast cell phenotype. Asthma, chronic obstructive pulmonary disease (COPD), and lung cancer are characterized by distinctive pathological changes in airway epithelial, SM, and mucous-generating cells. Acute and chronic pneumonitis and fibrosis occur in the alveolar space and interstitium with type 2 pneumocytes and interstitial fibroblasts/myofibroblasts playing a prominent role. At each site, inflammatory, immune, and vascular progenitor cells contribute to the injury and repair process. Many strategies have been used to investigate PKCs in lung injury. Isolated organ preparations and whole animal studies are powerful approaches especially when genetically engineered mice are used. More analysis of PKC isozymes in normal and diseased human lung tissue and cells is needed to complement this work. Since opposing or counter-regulatory effects of selected PKCs in the same cell or tissue have been found, it may be desirable to target more than one PKC isozyme and potentially in different directions. Because multiple signaling pathways contribute to the key cellular responses important in lung biology, therapeutic strategies targeting PKCs may be more effective if combined with inhibitors of other pathways for additive or synergistic effect. Mechanisms that regulate PKC activity, including phosphorylation and interaction with isozyme-specific binding proteins, are also potential therapeutic targets. Key isotypes of PKC involved in lung pathophysiology are summarized and current and evolving therapeutic approaches to target them are identified.     

Steroidal sapogenin contents in some domestic plants

In order to find out the values of the steroid resources for the future use. the compositions and contents of steroidal sapogenins from 13 domestic plants have been investigated. As a result,Dioscorea nipponica, D. quinqueloba andSmilax china were found to have large amount of diosgenin. And pennogenin inTrillium kamtschaticum andParis verticillata, yuccagenin inAllium fistulosum, hecogenin inAgave americana and neochlorogenin inSolanum nigum were appeared to be major steroidal sapogenins.