脑损伤后神经细胞凋亡变化及黄体酮保护作用的研究

目的：观察脑损伤后神经细胞凋亡变化及黄体酮的保护作用。方法雄性SD大鼠随机分为假手术组、脑损伤组和黄体酮治疗组,大鼠脑损伤模型制作是利用改良的Feeney自由落体损伤装置。各组于伤后24、48、72 h取材。用免疫组织化学法观察大鼠皮质和海马CA1区半胱氨酸天冬氨酸蛋白酶-3（Caspase-3）的表达水平变化。结果脑损伤组的Caspase-3阳性细胞数在皮质和海马CA1区较假手术组显著增加,黄体酮治疗组与脑损伤组比较,Caspase-3阳性细胞数明显减少,差异有统计学意义（P＜0.05）。结论黄体酮抑制脑损伤后Caspase-3的表达可能是黄体酮脑保护作用机制之一。     

大剂量维生素B6对大鼠重型脑外伤后神经细胞凋亡的影响

目的:观察和探讨大剂量维生素B6对大鼠外伤性脑损伤(TBI)后神经细胞凋亡的影响及意义。方法:54只SD雄性大鼠随机分为假手术组、脑损伤对照组,脑损伤大剂量维生素B6治疗组,12h、1d、3d、7d四个时间点,共9组,每组6只。采用Feeney,s自由落体打击法制作重型脑外伤模型,治疗组伤后维生素B6注射液450mg/KG腹腔注射,一天两次,对照组给予等量生理盐水。各时间点检测脑外伤后24h大脑伤灶周围皮层中B细胞淋巴瘤/白血病-2(bcl-2)的表达,原位缺口末端标记法(TUNEL)检测神经细胞的凋亡。结果:大剂量维生素B6治疗后,与脑外伤对照组相比较,1d、3d组可以上调bcl-2的表达,TUNEL阳性细胞减少,即凋亡细胞数减少,具有统计学意义(P0.05)。结论:TBI后应用大剂量维生素B6,能在部分时间段减少神经细胞凋亡,起到神经保护作用。     

反式白藜芦醇（TR）对痴呆大鼠海马旁回iNOS、caspase3表达的影响及机制

目的：观察反式白藜芦醇（TR）对痴呆大鼠海马旁回的作用。方法：大鼠随机分为给药组和对照组。在大鼠海马内注射Aβ25-35后5天,采用TUNEL法检测5组大鼠海马细胞及周围皮质凋亡情况,采用real time RT-PCR法检测两组大鼠海马及皮质iNOS、caspase-3表达。结果：对大鼠海马以及周边区域的iNOS、caspase-3的表达进行分析结果,给药组的iNOS、caspase-3表达水平下调2.59%,明显低于对照组的9.33%,差异有统计学意义。结论：TR对损伤的大鼠海马周围皮层神经元有保护作用,其机制可能部分与下调iNOS、caspase-3表达有关。     

辛伐他汀对创伤性脑损伤大鼠血清及海马NSE表达的影响

【摘要】目的通过观察辛伐他汀对创伤性脑损伤（TBI）大鼠神经元特异性烯醇化酶（NSE）在脑组织和血清中表达的影响,探讨辛伐他汀对大鼠TBI的治疗作用。方法8周龄的SD大鼠90只,随机分为假致伤组、对照组、治疗组,每组30只。对照组、治疗组参照改良的Feeney氏自由落体法制造TBI模型。治疗组大鼠于造模前晚及伤后每晚给予辛伐他汀10mg/kg灌胃;对照组及假致伤组同时给予等量淀粉灌胃。3组大鼠分别于伤后3h、12h、24h、3d、7d、14d取5只大鼠的颈总动脉血3mL,ELISA法测定NSE浓度;断头取脑,免疫组化法检测伤侧海马CA3区NSE表达。结果（1）ELISA法测定：对照组,伤后3h血清NSE即开始升高,24h至3d达高峰,14d仍高于假致伤和治疗组;治疗组,伤后3h血清NSE水平升高,24h达高峰,3d至7d下降,明显低于对照组,伤后14d接近假致伤组。（2）免疫组化检测：对照组伤后3h伤侧海马CA3区NSE光密度值下降,3d至7d达低谷,14d时仍显著低于假致伤组;治疗组的大鼠伤后3h伤侧海马CA3区NSE光密度值下降,12h至24h达低谷,但明显高于对照组,7d开始回升,14d时接近假致伤组水平。结论辛伐他汀可促进TBI后血清NSE水平的下降,提高损伤侧海马神经元的NSE表达,对TBI后大鼠损伤的神经元有保护作用。     

大鼠创伤性脑损伤后脑组织ICAM-1及血糖变化与细胞凋亡的实验研究

目的研究大鼠创伤性脑损伤后脑组织ICAM-1和血糖变化与神经细胞凋亡的关系。方法采用大鼠自由落体脑损伤模型（Feeney’s model），分别在伤后6、24、72、168h免疫组织化学检测ICAM-1、Hoechst染色定量凋亡细胞；测定伤前0．5h及伤后6、12、24、48、72、120、168h各组动物的血糖值。结果脑损伤程度越重，ICAM-1的IOD值、血糖浓度和神经细胞凋亡的个数升高的速度越快、峰值越高；神经细胞凋亡的个数从伤后6h即明显高于对照组。并持续增高。结论创伤性脑损伤后早期神经细胞凋亡可能与ICAM-1介导的细胞黏附及升高血糖的胰高血糖素、糖皮质激素等介导的细胞凋亡有关。     

亚低温促进大鼠颅脑创伤后海马新生 神经元长期存活及成熟

目的 探讨亚低温（MHT）对大鼠颅脑创伤（TBI）后海马齿状回区（DG）新生神经元长期存活及成熟的影 响。方法 59只健康成年SD大鼠随机分为假手术（sham）组、TBI组及TBI+MHT组,sham组15只,其余两组各22只。 TBI大鼠模型借助液压冲击脑损伤仪建立,打击压力设置为2.0 atm（1 atm=101.325 kPa）。TBI+MHT组大鼠在损伤后 立即接受MHT,降温后直肠目标温度为33.5 ℃,维持4 h,并于1.5 h内缓慢复温至37 ℃。在不同时间点应用Morris水 迷宫试验,mNSS评分比较大鼠神经功能恢复情况,免疫荧光染色等方法检测各组海马新生神经元长期存活及成熟 情况。结果 与TBI组比较,TBI+MHT组大鼠伤后4周的逃避潜伏期明显缩短,平台穿越次数及目标象限停留时间 均显著增加（P＜0.05）。损伤后1、2、4周TBI+MHT组大鼠较TBI组mNSS评分均降低（P＜0.01）。与sham组比较,损 伤后1、4、8周TBI组和TBI+MHT组大鼠损伤侧海马DG区BrdU阳性细胞数及BrdU/NeuN双阳性细胞数均增加（P＜ 0.05）,且TBI+MHT组较TBI组增加更明显（P＜0.05）。此外,与损伤后1周相比,损伤后4周及8周TBI组BrdU/NeuN 双阳性细胞数减少,而TBI+MHT组进一步增加（P＜0.05）。结论 MHT可促进TBI后新生神经元的长期存活及成 熟,促进TBI后神经功能恢复。     

雌激素对脑创伤大鼠海马神经细胞凋亡的干预

目的探讨雌激素在脑创伤后的作用。方法按照Marmarou建立大鼠创伤性损伤(TBI)模型,采用硫代巴比妥酸法测定氧自由基中间产物丙二醛(MDA),免疫组合法检测细胞凋亡相关基因Caspase-3的表达,原位末端标记(TUNEL)法检测细胞凋亡。结果雌激素治疗组海马CA1区MDA和Caspase-3及TUNEL阳性细胞均低于创伤组。结论大鼠创伤后雌激素通过抗氧化作用抑制神经细胞凋亡而起到脑保护作用。     

大鼠急性颅脑损伤后 IL-8在颅内表达的动态观察

目的：探讨大鼠急性颅脑外伤后颅内 IL-8的表达在继发性脑损伤中的作用及意义。方法将健康成年 SD 大鼠70只,随机分为对照组和损伤组后12、24、72、120 h,1周、2周组,每组10只。采用液压打击致伤法制作大鼠急性颅脑损伤模型。观察 IL-8在损伤灶周围的表达,并作脑组织含水量测量。结果损伤后12 h 时即可见坏组织死周围和皮层散在表达 IL-8的棕黄色阳性细胞,24 h 时阳性细胞明显增多达到高峰,不仅在神经元表达,也在胶质细胞、部分血管内皮细胞以及聚集成堆的中性粒细胞内表达;伤后72 h 阳性细胞开始减少,但经过光密度值检测仍高于对照组（P ＜0．05）。损伤后7 d 阳性细胞略有所回升,至损伤后14 d 阳性细胞明显减少,基本趋于平稳。损伤后病变侧各时间点阳性细胞数与对照组比较,差异均有统计学意义（P ＜0．05）。假手术组大鼠各时间点脑组织含水量差异均无统计学意义（P ＞0．05）。损伤组各时间点（除伤后14 d）脑含水量与假手术组相比均有增高（P ＜0．05）,在伤后24 h 达高峰,持续到72 h,伤后14 d 时脑含水量较对照组虽有所升高,但差异无统计学意义（P ＞0．05）。结论IL-8的表达与脑损伤后脑组织的病理生理学改变密切相关,在损伤早期 IL-8的高表达参与了脑水肿形成等继发性神经元损害过程,后期 IL-8的稍高表达可能与神经元修复有关。     

银杏叶提取物EGb761对大鼠脊髓损伤后的神经保护作用

目的探讨银杏叶提取物EGb761对实验性大鼠脊髓损伤后神经保护的作用及其机制。方法成年雄性SD大鼠132只,体重200～250 g,随机分为正常对照组（N组）、损伤组（T组）、甲基强的松龙治疗组（MP组）和EGb761治疗组（EGb761组）,每组33只。T组、MP组、EGb761组用改良Allen法以25GCF损伤力度致伤大鼠,建立T9脊髓中度损伤模型。术后4、8、24 h每组随机取3只动物切取损伤区1 cm脊髓节段,分别用黄嘌呤氧化酶法和硫代巴比妥酸（TBA）法测定脊髓组织中超氧化物歧化酶（SOD）活性和丙二醛（MDA）含量。分别于术后24 h、3、5、7、14 d处死动物（n=6）,快速取T9节段脊髓,TUNEL法标记细胞凋亡,免疫组化方法检测诱生型一氧化氮合酶（iNOS）的表达。结果术后4、8、24 h EGb761治疗组SOD活性及MDA含量与损伤对照组比较均差异有显著性（P〈0.01）。术后各时相点EGb761治疗组神经细胞凋亡指数和iNOS表达阳性细胞率均低于损伤对照组（P〈0.01或P〈0.05）。结论 EGb761能抑制脊髓损伤后的脂质过氧化反应,减轻神经细胞的凋亡,其机制可能与抑制iNOS表达有关。     

三七总皂甙对大鼠脊髓损伤后iNOS表达及细胞凋亡的影响

目的探讨大鼠脊髓损伤后应用三七总皂甙对诱导型一氧化氮合酶(iNOS)表达及细胞凋亡的影响。方法采用Allen’s法将64,只成年SD大鼠造成脊髓中度损伤模型,再随机分为损伤组和三七总皂甙组,分别给予生理盐水及三七总皂甙注射液治疗,损伤后不同时间点(1、3、7、14d)处死大鼠,用HE染色观察损伤脊髓组织病理变化,用免疫组化染色检测iNOS阳性细胞,原位末端标记法(TUNEL法)标记凋亡细胞。结果HE染色镜检发现脊髓组织病理学改变三七总皂甙组明显轻于损伤组;两组均发现iNOS表达及凋亡细胞,且损伤组iNOS表达及神经细胞凋亡指数均高于三七总皂甙组(P<0．05)。结论三七总皂甙注射液能抑制脊髓损伤后iNOS表达及神经细胞凋亡。     

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.     

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.     

Gender-related symptoms and correlates of alcohol dependence among men and women with a lifetime diagnosis of alcohol use disorders

This study explored gender-related symptoms and correlates of alcohol dependence in a crosssectional study of 150 men and 150 women with a lifetime diagnosis of alcohol use disorders (AUD). Participants were recruited in equal numbers from treatment settings, correctional centres and the general community. Standardized measures were used to determine participants' use of substances, history of psychiatric disorders and psychosocial stress, their sensation seeking and family history of substance use and mental health disorders. Multivariate analyses were used to detect patterns of variables associated with gender and the lifetime severity of AUD. Men had a longer history of severe AUD than women. Women had similar levels of alcohol dependence and medical and psychological sequelae as men, despite 6 fewer years of AUD. More women than men had a history of severe psychosocial stress, severe dependence on other substances and antecedent mental health problems, especially mood and anxiety disorders. There were differences in family history of alcohol-related problems approximating same-gender aggregation. The severity of a lifetime AUD was predicted by its earlier age at onset and the occurrence of other disorders, especially anxiety, among both men and women. The limitations in the generalizability of these findings due to sample idiosyncrasies are discussed.     

Biochemical and molecular characterisation of cubozoan protein toxins

Class Cubozoa includes several species of box jellyfish that are harmful to humans. The venoms of box jellyfish are stored and discharged by nematocysts and contain a variety of bioactive proteins that are cytolytic, cytotoxic, inflammatory or lethal. Although cubozoan venoms generally share similar biological activities, the diverse range and severity of effects caused by different species indicate that their venoms vary in protein composition, activity and potency. To date, few individual venom proteins have been thoroughly characterised, however, accumulating evidence suggests that cubozoan jellyfish produce at least one group of homologous bioactive proteins that are labile, basic, haemolytic and similar in molecular mass (42-46 kDa). The novel box jellyfish toxins are also potentially lethal and the cause of cutaneous pain, inflammation and necrosis, similar to that observed in envenomed humans. Secondary structure analysis and remote protein homology predictions suggest that the box jellyfish toxins may act as α-pore-forming toxins. However, more research is required to elucidate their structures and investigate their mechanism(s) of action. The biological, biochemical and molecular characteristics of cubozoan venoms and their bioactive protein components are reviewed, with particular focus on cubozoan cytolysins and the newly emerging family of box jellyfish toxins.     

Dose tolerability of chronically inhaled voriconazole solution in rodents

Invasive pulmonary aspergillosis (IPA) is a fungal disease of the lung associated with high mortality rates in immunosuppressed patients despite treatment. Targeted drug delivery of aqueous voriconazole solutions has been shown in previous studies to produce high tissue and plasma drug concentrations as well as improved survival in a murine model of IPA. In the present study, rats were exposed to 20 min nebulizations of normal saline (control group) or aerosolized aqueous solutions of voriconazole at 15.625 mg (low dose group) or 31.25 mg (high dose group). Peak voriconazole concentrations in rat lung tissue and plasma after 3 days of twice daily dosing in the high dose group were 0.85 ± 0.63 μg/g wet lung weight and 0.58 ± 0.30 μg/mL, with low dose group lung and plasma concentrations of 0.38 ± 0.01 μg/g wet lung weight and 0.09 ± 0.06 μg/mL, respectively. Trough plasma concentrations were low but demonstrated some drug accumulation over 21 days of inhaled voriconazole administered twice daily. Following multiple inhaled doses, statistically significant but clinically irrelevant abnormalities in laboratory values were observed. Histopathology also revealed an increase in the number of alveolar macrophages but without inflammation or ulceration of the airway, interstitial changes, or edema. Inhaled voriconazole was well tolerated in a rat model of drug inhalation.