层黏连蛋白与脑微血管内皮细胞对共同植入缺血模型大鼠脑内神经干细胞增殖与凋亡的调节

目的:将由神经干细胞、层黏连蛋白、脑微血管内皮细胞构建的共移植体植入缺血模型鼠大脑内,观察共移植体中神经干细胞的增殖和凋亡方法:实验于1996-01/1996-12在佳木斯大学神经科学所完成实验材料:同一基因背景Wistar大鼠,体质量300～350g,雄性,6月龄,购自哈尔滨医科大学实验动物学部实验方法:①取出生24h内Wistar新生鼠,培养神经干细胞②将传代脑微血管内皮细胞消化后,以3×107L-1密度接种于培养瓶中,12h贴壁后吸出脑微血管内皮细胞培养液,涂以一层细胞外基质后,接种经0.125%胰酶消化20min后吹打成单细胞的神经干细胞悬液,密度为6×108L-1,加入神经干细胞完全培养液培养3～6h,镜下观察神经干细胞完全贴壁后,吸出培养液,再涂以一薄层细胞外基质,接种以上相同密度脑微血管内皮细胞,加入神经干细胞完全培养液共培养12～24h,镜下观察脑微血管内皮细胞利用免疫荧光方法检测共移植体③制备大鼠脑缺血模型模型制备完成后大鼠虽然有脑缺血症状,但伴下列情形之一的不纳入:神经学症状评分低于2分;;取脑时发现蛛网膜下腔出血;;脑组织石蜡切片苏木精-伊红染色无缺血病理改变;;未到观察时相点便死亡的共48只大鼠模型制作成功将Wistar大鼠随机分为神经干细胞组、共移植体组,每组24只2组分4个时间点3,7,14,60d,每个时间点6只模型建立后3d,神经干细胞组移植神经干细胞,共移植体组移植共移植体,各5μL移植前神经干细胞经Brdu5mmol/L标记3d,再将其溶解于PBS中,细胞密度调整为2×1011L-1,移植部位为右侧纹状体区通过免疫组织化学和免疫荧光方法观察神经干细胞增殖和凋亡情况结果:48只大鼠均进入结果分析①神经干细胞增殖情况:细胞数3d开始增加,14d达高峰,60d减少两组3,7,14,60d细胞数相比,差异显著(神经干细胞组:19.57±4.27,22.25±4.53,39.13±4.52,7.13±2.75;共移植体组:39.88±4.26,40.50±4.03,56.88±9.33,17.13±4.09,P<0.05,P<0.01)。3,7d细胞增殖数较稳定,Brdu阳性细胞主要集中在针迹部位,14dBrdu阳性细胞最多,散在分布,部分与周围组织整合,60dBrdu阳性细胞细胞数下降,大部分已与周围组织整合。②神经干细胞凋亡率情况:随着时间延长,共移植体组细胞凋亡率降低,与神经干细胞组相比,差异显著[单纯神经干细胞组3,7,14,60d分别为:(35.57±1.85)%,(28.58±1.26)%,(17.86±1.32)%,(2.05±0.67)%;共移植体组分别为:(24.77±2.44)%,(22.75±3.40)%,(9.88±1.81)%,(0.75±0.32)%,P<0.05,P<0.01]。结论:层黏连蛋白和脑微血管内皮细胞与神经干细胞共移植体移植入缺血模型大鼠脑内可明显促进神经干细胞增殖,减少其凋亡。     

首乌丹参方预处理对大鼠心肌缺血再灌注损伤蛋白激酶C的影响

目的:分析首乌丹参方预处理对大鼠心肌缺血再灌注损伤的保护机制是否通过促进蛋白激酶CmRNA水平表达发挥作用。方法:实验于2003-07/2004-05在天津中医药大学中医药研究中心省部共建教育部方剂学重点实验室完成。SPF级雄性Wistar大鼠,体质量(336±20)g。随机数字表法分为7组:假手术组(取血清作正常对照)、模型组、首乌丹参方高剂量组、(9g/kg)、首乌丹参方中剂量组(4.5g/kg)、首乌丹参方低剂量组(2.25g/kg)、复方丹参滴丸阳性对照组(4.5g/kg)、消心痛对照组(1.67mg/kg),每组12只。首乌丹参方浸膏,由天士力药业集团提供,100g生药出膏15.26g(每克浸膏的生药量为6.55g),批号:20020318,给药剂量分别为9,4.5,2.25g/kg,实验前称取一定量的浸膏,用0.5%羧甲基纤维素钠分别配成0.9,0.45,0.225g/mL浓度的混悬液,每克生药加天然冰片5.19mg。阳性对照药复方丹参滴丸浸膏,黑褐色,由天士力药业集团提供,100g生药出膏21.47g(每克浸膏的生药量为4.66g,批号:20020423)。给药剂量以生药计算为4.5g/kg,每克生药加合成冰片6.15mg;阳性对照药消心痛(硝酸异山犁酯)片,每片含消心痛5mg,天津太平洋制药有限公司生产(批号:20040302),药液浓度0.167mg/mL,配制方法同上。采用结扎大鼠冠状动脉前降支30min/开放120min建立心肌缺血再灌注损伤模型。假手术组只穿线不结扎,30min时将所穿线迅速拎起后再放下,快速关胸。造模后假手术组、模型组给0.5%羧甲基纤维素钠液,余组将药物用0.5%羧甲基纤维素钠配制成混悬液,给药体积皆为1mL/100g。通过RT-PCR分子生物学方法观察蛋白激酶CmRNA表达情况以及首乌丹参方预处理对其的影响。结果:84只大鼠均进入结果分析。与假手术组相比,模型组和首乌丹参方各组蛋白激酶CmRNA表达均有所下降;与模型组相比,首乌丹参方中剂量组、复方丹参滴丸阳性对照组和消心痛对照组表达上调,差异均有显著性意义,首乌丹参方高剂量组及低剂量组也能促进蛋白激酶CmRNA的表达,但差异没有显著性意义。结论:首乌丹参方预处理可促进缺血再灌注损伤大鼠心肌蛋白激酶C表达。首乌丹参方可能通过激动蛋白激酶C对缺血再灌注损伤的心肌起保护效应。     

山东泰安地区两个自然村人群血脂状况调查

目的:调查了解山东泰安地区农村人群的血脂水平,为该地区农民心脑血管疾病的预防提供基础数据。方法:实验于2006-06随机选取泰安市两个自然村650人进行健康检查,男270人,女380人,年龄2～88岁。①650人均于早晨空腹采集静脉血3mL,静置凝固后,2h内分离血清,采用全自动生化分析仪常规测定患者血脂指标总胆固醇、三酰甘油水平。②总胆固醇>5.2mmol/L判定为高胆固醇血症,三酰甘油>1.7mmol/L判定为高三酰甘油血症。结果:650人全部完成血样采集与血脂指标检测。①不同性别血脂水平比较:被检人群男性三酰甘油水平显著高于女性[(1.16±1.47),(0.95±0.84)mmol/L,P<0.05];男女总胆固醇水平基本相似(P>0.05)。②不同年龄段血脂水平比较:8人因具体年龄段不祥被排除,剩余642人分为3个年龄段:2～30岁、31～50岁、51～88岁。结果31～50岁人群三酰甘油水平显著高于2～30岁人群[(1.14±1.47),(0.88±0.70)mmol/L,P<0.05],略高于51～88岁人群(P>0.05);51～88岁人群总胆固醇水平>31～50岁人群>2～30岁人群,相邻年龄段间比较差异均有显著性意义(P<0.001)。③各年龄段男女间血脂水平比较:2～30岁人群和51～88岁人群三酰甘油水平性别间差异均无显著性意义(P>0.05),但总胆固醇水平女性均显著高于男性[(2.38±1.47),(1.85±1.26)mmol/L,P<0.05;(4.48±0.92),(4.18±0.76)mmol/L,P0.01]。31～50岁人群男性三酰甘油水平显著高于女性[(1.51±2.06),(0.90±0.80)mmol/L,P<0.01],总胆固醇水平基本相似(P>0.05)。④各年龄段男女高血脂症构成比:31～50岁人群高三酰甘油血症构成比显著高于2～30岁、51～88岁人群(23.1%,5.5%,9.3%,P<0.01);随着年龄增高,各年龄段高胆固醇血症构成比逐渐升高,相邻年龄段间差异均有显著性意义(2.7%,8.6%,15.6%,P<0.01)。31～50岁人群男性高三酰甘油血症、高胆固醇血症的构成比显著高于女性(P<0.01);2～30岁、51～88岁人群男女间高血脂症的构成比基本相似(P>0.05)。结论:①山东泰安地区农村人群高三酰甘油血症,31～50岁年龄段患病率最高;高胆固醇血症患病率随年龄增长逐步上升,提示中年人健康体检尤为重要。②高脂血症应从青少年开始加大预防和保健力度,进行广泛和反复的健康教育,倡导健康的生活方式和生活行为。     

藏族糖尿病患者代谢功能改善与面对面教育的效应

目的:观察面对面随访方式的糖尿病教育对藏族患者代谢功能改善的作用。方法:①选择2005-08/2006-08在西藏军区总医院心内科住院和门诊就诊的糖尿病患者113例。男69例,女44例;年龄48～71岁。纳入糖尿病患者均符合1999年世界卫生组织糖尿病诊断标准,且对检测项目知情同意;均为离解放军西藏军区总医院最近的娘热乡藏族糖尿病患者。②测量身高、体质量、收缩压、舒张压,计算体质量指数=体质量/身高2。采用日立7600型全自动生化分析仪测定血清总胆固醇、三酰甘油、高密度脂蛋白胆固醇、低密度脂蛋白胆固醇水平。采用TosohHLC-723GHbV型高压液相色谱仪测定糖化血红蛋白。③所有参试者得到一份本院编写的藏汉对照糖尿病教育手册,并在分组前统一讲解,以上大课的形式进行普及教育。根据每个患者不同情况,由内分泌专业医师给予不同处方,包括饮食、用药、生活方式调整。随后将患者随机分为教育组56例(男35例,女21例)和对照组57例(男34例,女23例)。对照组主要采用患者主动咨询(电话咨询或医院复查)和每一两个月一次普及教育的方式,但医生不采用面对面主动干预;教育组的干预方式除上述普及教育以外,每2周进行一次面对面、一对一随访,并及时调整处方。半年后比较两组的代谢指标差异。④计量和计数资料的差异比较分别采用方差分析和χ2检验。结果:藏族糖尿病患者113例均完成随访并进入结果分析。①组间比较:干预前两组体质量指数、收缩压、舒张压、总胆固醇、三酰甘油、高密度脂蛋白胆固醇、低密度脂蛋白胆固醇、糖基化血红蛋白比较,差异不明显(P>0.05)。教育半年后教育组的体质量指数、收缩压、舒张压、总胆固醇、三酰甘油、低密度脂蛋白胆固醇和糖化血红蛋白均明显低于对照组(t=2.0156～4.7592,P<0.05～0.01),高密度脂蛋白胆固醇明显高于对照组(t=3.8031,P<0.01),其中以糖基化血红蛋白差异最明显。②组内比较:教育组糖尿病教育半年后血清总胆固醇、三酰甘油、低密度脂蛋白胆固醇和糖化血红蛋白均较干预前明显降低(t=1.9832～3.6446,P<0.05～0.01),高密度脂蛋白胆固醇较干预前升高(t=2.8219,P<0.01),教育干预前后体质量指数无明显变化(P>0.05)。对照组糖尿病教育半年后,体质量指数、总胆固醇、三酰甘油、低密度脂蛋白胆固醇和糖化血红蛋白、高密度脂蛋白胆固醇与教育前相近(P>0.05)。结论:藏族糖尿病患者采取面对面的随访方式进行糖尿病教育较一般形式的糖尿病教育更好的改善其代谢功能。     

Extract of the Chinese herbal formula Huo Luo Xiao LingDan inhibited adjuvant arthritis in rats

Ethnopharmacological relevance

The herbal formula Huo Luo Xiao Ling Dan (HLXL) and its modifications have been used in traditional Chinese medicine for about one hundred years to alleviate pain and inflammation.

Aim

To investigate the effects of HLXL on complete Freund's adjuvant (CFA)-induced multiple-joint arthritis in rats.

Materials and methods

Male Lewis rats, 190–210 g, were immunized subcutaneously at the base of the tail with 200 μl of heat-killed Mycobacterium tuberculosis in mineral oil (5 mg/ml). HLXL (2.30 and 4.60 g/kg) or vehicle control (n = 8 per group) was administered orally (i.g.) once a day between days 16 and 25 post-CFA injection. The rats were observed for signs of arthritis with arthritic changes (erythema, edema, induration) being scored on a scale of 0–4 of increasing severity using a standard scoring system. The maximum arthritis score per rat was 16. A plethysmometer was used to measure edema volume in each paw. Adverse effects of HLXL were monitored by closely observing the animals for unusual behavioral changes. Levels of tumor necrosis factor alpha (TNF-α) and interleukin-1 beta (IL-1β) in local tissue were measured by enzyme-linked immunosorbent assay on day 25 post-CFA.

Results

HLXL significantly decreased arthritis scores between days 23–25 in the 2.30 g/kg group and 21–25 in the 4.60 g/kg group (p < 0.05). It reduced paw edema on days 22 and 24 in the 2.30 g/kg group and on days 20, 22 and 24 in the 4.60 g/kg group compared to control (p < 0.05). Local tissue TNF-α and IL-1β levels on day 25 post-CFA injection were significantly (p < 0.05) lower in rats treated with HLXL than in control rats. No observable adverse effects were found.

Conclusion

The data suggest that HLXL produces significant anti-arthritic effects that may be mediated by suppressing pro-inflammatory cytokines, and it appears to be safe.     

Primary Breast Angiosarcoma: A Retrospective Study of 36 Cases from a Single Chinese Medical Institute with Clinicopathologic and Radiologic Correlations

The aim of this study was to present our experience with primary breast angiosarcoma (PBA) by describing a large series of cases with an emphasis on clinicopathologic and radiologic correlations. Thirty‐six cases of PBA diagnosed at our institution between 2006 and 2014 were retrospectively evaluated. All but one case occurred in women with a median age of 35.5 years. The majority of patients presented with a deeply located painless mass, whereas a minority manifested as diffuse enlargement or swelling of the breast. Magnetic resonance imaging showed poorly demarcated lesions with low signal intensity on T1‐weighted images, markedly high intensity on T2‐weighted images, and prolongation of enhancement upon dynamic study. Histologically, 19 cases (52.8%) were low grade, 12 cases (33.3%) were intermediate grade, and 5 cases (13.9%) were high grade. Follow‐up information was available for 27 patients and revealed local recurrence and/or metastasis in 16 patients (59.3%). Five patients (18.5%) died of the disease at a median interval of 20 months. Univariate analysis showed that tumor differentiation had effect on disease‐free survival (DFS) (p = 0.005) but failed to predict overall survival (OS) (p = 0.645). The treatment modality was related to OS (p = 0.042) but not DFS (p = 0.131). The Cox proportional hazards regression model suggested that tumor differentiation was an independent predictor of DFS (p = 0.015). We hypothesize that tumor differentiation may be used as a prognostic factor for this rare malignancy. Clinicopathologic and radiologic correlation may help pathologists to arrive at the correct diagnosis of PBA.     

Introducing seasonal influenza vaccine in low‐income countries: an adverse events following immunization survey in the Lao People's Democratic Republic

Objective

In 2012, Lao PDR introduced seasonal influenza vaccine in pregnant women, persons aged ≥50 years, persons with chronic diseases, and healthcare personnel. We assessed adverse events following immunization (AEFI).

Methods

We used a multistage randomized cluster sample design to interview vaccine recipients.

Findings

Between April and May 2012, 355 902 were vaccinated. Of 2089 persons interviewed, 261 (12·5%) reported one or more AEFI. The most commonly reported AEFIs were local reactions. No hospitalizations or deaths were reported; 16% sought medical care. Acceptance and awareness of vaccination were high.

Conclusions

Following the introduction of seasonal influenza vaccine in Lao PDR, self-reported adverse events were mild.     

Thrombotic occlusion of the common carotid artery (CCA) in acute ischemic stroke treated with intravenous tissue plasminogen activator (TPA)

Although common carotid artery (CCA) occlusions are rare, acute clinical presentations vary from mild to devastating strokes primarily due to tandem occlusions in the intracranial arteries. Three patients with acute CCA occlusions were treated with systemic tissue plasminogen activator (TPA). Blood pressures were kept at the upper limits allowed with TPA therapy with fluid balance and the 'head-down' position. Recanalization occurred in intracranial vessels only. Marked early neurological improvement occurred in two of three patients. CCA occlusions should not be considered contra-indication to systemic thrombolysis.     

Human brain imaging of nicotinic acetylcholine α4β2* receptors using [18F]Nifene: Selectivity,functional activity,toxicity, aging effects,gender effects,and extrathalamic pathways

Nicotinic acetylcholinergic receptors (nAChR's) have been implicated in several brain disorders, including addiction, Parkinson's disease, Alzheimer's disease and schizophrenia. Here we report in vitro selectivity and functional properties, toxicity in rats, in vivo evaluation in humans, and comparison across species of [¹⁸F]Nifene, a fast acting PET imaging agent for α4β2* nAChRs. Nifene had subnanomolar affinities for hα2β2 (0.34 nM), hα3β2 (0.80 nM) and hα4β2 (0.83 nM) nAChR but weaker (27–219 nM) for hβ4 nAChR subtypes and 169 nM for hα7 nAChR. In functional assays, Nifene (100 μM) exhibited 14% agonist and >50% antagonist characteristics. In 14‐day acute toxicity in rats, the maximum tolerated dose (MTD) and the no observed adverse effect level (NOAEL) were estimated to exceed 40 μg/kg/day (278 μg/m²/day). In human PET studies, [¹⁸F]Nifene (185 MBq; <0.10 μg) was well tolerated with no adverse effects. Distribution volume ratios (DVR) of [¹⁸F]Nifene in white matter thalamic radiations were ～1.6 (anterior) and ～1.5 (superior longitudinal fasciculus). Habenula known to contain α3β2 nAChR exhibited low levels of [¹⁸F]Nifene binding while the red nucleus with α2β2 nAChR had DVR ～1.6–1.7. Females had higher [¹⁸F]Nifene binding in all brain regions, with thalamus showing >15% than males. No significant aging effect was observed in [¹⁸F]Nifene binding over 5 decades. In all species (mice, rats, monkeys, and humans) thalamus showed highest [¹⁸F]Nifene binding with reference region ratios >2 compared to extrathalamic regions. Our findings suggest that [¹⁸F]Nifene PET may be used to study α4β2* nAChRs in various CNS disorders and for translational research.