DSA技术在恒河猴脑脊液动态采集中应用

探索数字减影血管造影(digital subtraction angiography,DSA)插管技术在恒河猴(Macaca mulatta)脑脊液动态采集中的应用。将28只恒河猴作为实验动物,随机分为实验组和对照组,每组14只,经左前肢静脉推注3%戊巴比妥钠溶液全麻后,常规剔除背部脊柱周围体毛、消毒,实验组从穿刺开始就在DSA技术下行腰椎间隙穿刺,对照组直接行腰椎间隙穿刺,将硬膜外麻醉导管经腰椎蛛网膜下腔送至枕大池并埋置泵。对不同穿刺方法的结果数据进行χ2检验;神经功能参照改良Tarlov评分标准,切口采用改良的切口愈合等级评分标准。实验组经腰椎间隙穿刺蛛网膜下腔全部成功;对照组8只穿刺成功,6只穿刺失败;所有动物埋置泵均能从泵中采集到无色透明的脑脊液;术后恒河猴均于当天恢复正常。在DSA技术下经腰椎间隙穿刺蛛网膜下腔并埋置泵,成功率高,操作简便、安全性好,可行性强;通过埋置的泵采集脑脊液,快速省时,可实现在动物清醒时、全生理状态下对脑脊液进行动态多次采集,有助于提高非人灵长类实验动物的福利,由于采样导管置于枕大池,采集到的脑脊液样品质量有保证,可用于研究缺血性脑卒中发生机制,满足检验需要,具有一定的实用价值。     

家兔自发性蛛网膜下腔出血早期脑损伤模型的初步研究

目的:建立一种可重复的、有典型神经损伤症状的自发性蛛网膜下腔出血(SAH)动物模型,为研究蛛网膜下腔出血后早期脑损伤(EBI)的机制提供可靠的动物模型。方法:选用新西兰大耳白兔60只,随机分为实验组、对照组、空白组。采用枕大池穿刺一次注入自体动脉血法建立SAH模型。在4h、8h、12h、24h、48h、72 h时间点,观察比较行为学与脑组织形态学的变化。结果:(1)实验组枕大池和蛛网膜下腔内发现大量的血凝块和血液。(2)实验组光镜下可观察到蛛网膜下腔大量红细胞,神经元细胞水肿,电镜下可见胶质细胞空泡样改变,神经元细胞线粒体肿胀,髓鞘内存在空泡和板层分离现象。(3)实验组造模后均出现明显的神经系统损伤,对照组4h出现典型神经系统损伤表现,12h后恢复正常,空白组未见神经系统损伤表现。结论:枕大池一次注血法是一种简便、可重复的症状性自发性蛛网膜下腔出血早期脑损伤模型,适宜于研究早期脑损伤的致病机制。     

家兔自发性蛛网膜下腔出血早期脑损伤模型的初步研究

目的：建立一种可重复的、有典型神经损伤症状的自发性蛛网膜下腔出血（sAH）动物模型,为研究蛛网膜下腔出血后早期脑损伤（EBI）的机制提供可靠的动物模型。方法：选用新西兰大耳白兔60只,随机分为实验组、对照组、空白组。采用枕大池穿刺一次注入自体动脉血法建立SAH模型。在4h、8h、12h、24h、48h、72h时间点,观察比较行为学与脑组织形态学的变化。结果：（1）实验组枕大池和蛛网膜下腔内发现大量的血凝块和血液。（2）实验组光镜下可观察到蛛网膜下腔大量红细胞,神经元细胞水肿,电镜下可见胶质细胞空泡样改变,神经元细胞线粒体肿胀,髓鞘内存在空泡和板层分离现象。（3）实验组造模后均出现明显的神经系统损伤,对照组4h出现典型神经系统损伤表现,12h后恢复正常,空白组未见神经系统损伤表现。结论：枕大池一次注血法是一种简便、可重复的症状性自发性蛛网膜下腔出血早期脑损伤模型,适宜于研究早期脑损伤的致病机制。     

26例新西兰白兔正常骨髓像观察

检测了第四军医大学实验动物中心提供的 2 6只成年健康新西兰白兔的骨髓像 ,对国内不同品种兔骨髓像资料加以补充和丰富。1　材料和方法　按照施新猷所著医学动物实验方法中健康兔的判断标准 ,取 2 6只健康新西兰白兔 ,年龄 8～ 18个月 ,体重1 85～ 3 4 5ｋｇ。常规 3 %戊巴比妥钠 ( 3 0ｍｇ ｋｇ)腹腔内麻醉 ,剪去术区毛发 ,无菌条件下在每只兔左侧胫骨前内侧骨面距胫骨平台 (即胫骨近侧关节面 ) 0 8～ 1 5ｃｍ范围内用骨穿针穿刺抽取 0 2～ 0 4ｍｌ不抗凝骨髓 ,立即涂片 3张做瑞氏染色 ,油镜下计数 5 0 0个有核细胞 ,并鉴定细胞种类及…     

兔VX2肝癌模型制作改良及影像学评价

目的 对兔VX2 肝癌模型制作进行改良,以用于介入治疗学研究,同时探讨瘤灶的CT 表现及CT 在检测瘤灶中的作用.方法 将VX2瘤细胞接种于兔皮下使其成瘤并传代;新西兰兔24只,以改良嵌插法建立移植性肝癌模型,于建模后7、14、21 d分别行超声、CT及血管造影检查,用于检测兔肝VX2 瘤灶,评估瘤灶生长变化;随后处死动物,进行尸解,评估影像检查结果.结果 24只（100%） 动物以改良嵌插法建立移植性肝癌模型全部成功.瘤灶以种植后2周CT显示最清楚和典型,直径1 cm～2 cm 左右,平扫呈低密度或等密度,动脉早期明显强化,门脉期呈低密度,与周围肝组织分界较清楚.肝动脉造影显示肿瘤富血供.而种植后超过3 周的肿瘤大部分发生坏死.结论 嵌插改良法是一种值得推广的建立移植性肝癌模型的方法;在对瘤灶进行影像学评价上应尽量选择CT检查,接种后1 周左右的瘤灶较小而难以观察;2 周左右呈肝动脉源性血供丰富的约1 cm～2 cm的实体瘤,造影征像为肿瘤血管与肿瘤染色;3 周以上瘤灶大多出现明显示坏死;因此对1～2cm大小的兔VX2 肝癌瘤体,最适合行血管造影检查.     

兔有创动脉血压监测的新方法

目的建立一种经兔耳正中动脉穿刺置管进行动脉血压监测新方法。方法健康家兔16只,雌雄不分,体重2．0±0．3k。动物随机分成两组,颈内动脉穿刺置管组（A组）8只,耳正中动脉穿刺置管组（B组）8只。两组均用22G动静脉留置针行穿刺置管后接有创动脉换能器套件联接监护仪监测有创动脉血压的变化,实验后家兔正常饲养7d,观察其死亡率。结果有创动脉压：A组和B组两组差异无统计学意义（P〉0．05）;死亡率：A组明显高于B组（P〈0．05）。结论经兔耳正中动脉穿刺置管进行有创动脉血压监测比颈内动脉置管法要好,是一种简单、有效的方法,死亡率低,可以在兔的动物实验中推广。     

液流径向变化对尾巨桉单株日蒸腾量估算的影响

采用Granier热扩散方法(TDP)测定了尾巨桉2个径向深度(0～2和2～4 cm)的液流速率,探讨液流径向变化对树木蒸腾估算产生的影响、不同个体大小的估算误差以及估算误差与光合有效辐射(PAR)和水汽压亏缺(VPD)的关系.结果表明: 尾巨桉的日蒸腾量在单点径向深度0～2 cm条件下估算值最大,两点综合估算值次之,单点径向深度2～4 cm最小,单点径向0～2 cm和径向2～4 cm所得的日蒸腾量分别较两点综合估算值高估了32.9%和低估了58.7%;用单点径向0～2 cm估算蒸腾量时所产生的误差随胸径的递减而减少,胸径为17.7、12.9和9.8 cm的树木产生的估算误差分别为51.7%、33.0%、18.0%,而单点径向2～4 cm所产生的估算误差则没有明显的径级差异;PAR和VPD都与单点径向0～2 cm估算误差有较好的正相关性,相比较VPD而言,PAR与单点径向0～2 cm估算误差的相关性更好.相比传统的仅用单一深度液流速率估算单株蒸腾耗水量的方法,两点估算方法一定程度上提高了日蒸腾量计算的精度,为尾巨桉人工林森林耗水量估算提供了可靠依据.     

经皮穿刺抽取大鼠脑脊液方法的改良

目的: 改进传统经皮穿刺抽取脑脊液的方法,提高采集大鼠脑脊液动物存活率。方法: 90只SD大鼠随机分为对照组、传统穿刺组和改良穿刺组(每组30只)。对照组不进行穿刺。传统穿刺组按反握抓持法进行穿刺取样。改良穿刺组采用左手固定大鼠头部,将其抬高至约135°,右手正握改良1 ml注射器,以枕骨隆突和第一颈椎为标志定位枕骨大孔,在枕骨大孔中心偏下平行进针,抽取脑脊液。分别于第1、4、7日抽取大鼠脑脊液,每次抽取记录取样时间、采集量、脑脊液性状,抽取后3 d观察大鼠存活情况,选取存活大鼠进行重复采样。结果: 传统经皮穿刺取样法采样成功率为63.33%,三次重复采样动物存活率为10%;改良经皮穿刺取样法采样成功率为86.67%,三次重复采样动物存活率为50%,相比传统采样有显著升高(P＜0.01)。结论: 改良经皮穿刺取样法操作简单,耗时少,成功率高。操作无需定位设备,单人可以完成。动物反复取样存活率高,方便进行脑脊液重复采样。     

不同造影方法显示弹性酶诱导的兔囊状动脉瘤

目的评估经股动脉穿刺超选造影、静脉造影、心脏穿刺造影和左侧耳中央动脉穿刺造影方法显示弹性酶诱导的兔囊状动脉瘤的可行性。方法取10只新西兰兔,采用弹性酶诱导方法制作兔右侧颈总动脉起始部囊状动脉瘤模型。术后3周对所有动物分别进行经股动脉穿刺超选造影、静脉造影、心脏穿刺造影和左侧耳中央动脉穿刺造影,根据造影结果测量动脉瘤短径、长径和瘤颈宽。采用重复测量设计的方差分析方法比较不同造影方法显示的动脉瘤相关参数测量结果差异。结果采用经股动脉穿刺超选造影、静脉造影、心脏穿刺造影和左侧耳中央动脉穿刺造影方法均能较清楚的显示的动脉瘤,比较不同造影方法测量的动脉瘤短径、长径和瘤颈宽均无统计学差异（P值分别是0.646,0.427和0.625）。结论不同的造影方法均能较准确显示弹性酶诱导的兔囊状动脉瘤大小,根据不同的实验目的和条件可选用不同的造影方法。     

青海门源地区大(狂鸟)和雕鸮的食性比较

1999～2002年的6～8月份,在青海门源地区收集了大(狂鸟)和雕鸮的吐弃块(pellets)和残留食物(food remains),带回实验室进行分检鉴定、研究分析.大(狂鸟)食物中共有736个猎物,其中高原鼢鼠28只、高原鼠兔139只、甘肃鼠兔142只、田鼠科动物422只、雀形目鸟类4只、香鼬1只;各猎物对大(狂鸟)食物的生物量贡献率分别为14.26%、40.79%、17.39%、26.99%、0.22%、0.35%.雕鸮食物中共有330个猎物,其中高原鼢鼠17只、高原鼠兔77只、甘肃鼠兔44只、田鼠科动物183只、雀形目鸟类2只、红脚鹬2只、高原兔5只;各猎物对雕鸮食物的生物量贡献率分别为11.83%、30.87%、7.36%、16.00%、0.15%、0.62%、33.17%.雕鸮的食物生态位宽度与大(狂鸟)的食物生态位宽度相近,食物生态位高度重叠,但是它们捕食同种猎物的比例显著不同.     

Calculating lumbar puncture depth in children

Lumbar puncture was performed in 195 children and the depth of needle was recorded. Our results show that the depth of lumbar puncture necessary to obtain uncontaminated cerebrospinal fluid correlates best with the child's weight. The simple formula: mean depth of insertion (cm) = 1.3 + 0.07 x body weight (kg), can be used to estimate the depth of lumbar puncture of children older than 3 months. The depths of lumbar puncture of children younger than 3 months are mostly 1.0-1.5 cm.     

Cerebrospinal fluid sampling from guineapigs: sample volume--related changes in protein concentration in control animals and animals in the relapsing phase of chronic relapsing experimental allergic encephalomyelitis

Cerebrospinal fluid (CSF) was removed from guineapigs by puncture of the cisterna magna and the total sample volume of 200-360 microl divided into 40 microl aliquots. After determination of albumin and IgG in these CSF aliquots it was found that successive samples gave different results. In general, up to 100 microl CSF could be removed before the protein concentration began to increase. In animals with chronic relapsing experimental allergic encephalomyelitis (CR-EAE) the rise in albumin concentration was accompanied by a corresponding fall in the number of white cells in later samples.     

A technique for repeated sampling of pure cerebrospinal fluid from the conscious rabbit

For the study of drug pharmacokinetics' a reliable method for repeated sampling of uncontaminated cerebrospinal fluid (CSF) from a conscious rabbit was established. The subdural space was punctured between a spinous processes of the lumbar vertebrae. A catheter was inserted through a cannula and pushed up to the region of the cisterna magna. After removing the cannula, the distal end of the catheter was sealed and placed subcutaneously until the onset of the experiments. Ten days after the insertion, samples of clear and colorless CSF up to 1 ml could be withdrawn.     

Non‐invasive sentinel lymph node mapping and needle guidance using clinical handheld photoacoustic imaging system in small animal

Translating photoacoustic imaging (PAI) into clinical setup is a challenge. Handheld clinical real‐time PAI systems are not common. In this work, we report an integrated photoacoustic (PA) and clinical ultrasound imaging system by combining light delivery with the ultrasound probe for sentinel lymph node imaging and needle guidance in small animal. The open access clinical ultrasound platform allows seamless integration of PAI resulting in the development of handheld real‐time PAI probe. Both methylene blue and indocyanine green were used for mapping the sentinel lymph node using 675 and 690 nm wavelength illuminations, respectively. Additionally, needle guidance with combined ultrasound and PAI was demonstrated using this imaging system. Up to 1.5 cm imaging depth was observed with a 10 Hz laser at an imaging frame rate of 5 frames per second, which is sufficient for future translation into human sentinel lymph node imaging and needle guidance for fine needle aspiration biopsy.      

不同土层深度配施缓释/普通尿素对土壤氮素和酶活性及玉米产量的影响

通过2017—2018两年田间试验,研究了不同土层深度配施缓释(PCU)/普通尿素(PU)对0～30 cm土层土壤无机氮含量、酶活性和玉米产量的影响。试验设置不施氮肥(CK)、普通尿素一次施肥(PU₁,5～10 cm土层)、普通尿素传统两次施肥(PU₂,5～10 cm土层,60%种肥+40%追肥)、普通尿素一次分层施肥(PU₃,5～10 cm土层20%N+15～20 cm土层30%N+25～30 cm土层50%N)、不同土层深度缓释/普通尿素配施[PCU₁～PCU₄,均为5～10 cm土层20%N(普通尿素)+15～20 cm土层30%N(配施)+25～30 cm土层50%N(配施),其中PCU₁～PCU₄的15～20和25～30 cm土层PCU:PU分别为3:7、3:7,5:5、5:5, 3:7、5:5, 5:5、3:7]共8个处理。结果表明: 与CK相比,PU₁能够满足玉米生育前期对0～10 cm土层氮素的需求,PU₂和PU₃能够满足玉米发育前期对10～30 cm土层氮素的需求,不同土层深度配施缓释/普通尿素能够满足玉米整个生育时期对氮素的需求。与PU₁～PU₃相比,不同土层深度配施缓释/普通尿素可显著增加灌浆期和成熟期10～20和20～30 cm土层NO₃^--N、NH₄⁺-N、碱解氮含量和脲酶、蛋白酶活性。与PU₃相比,不同土层深度配施缓释/普通尿素处理2017和2018年玉米产量分别提高2.3%～24.6%和1.3%～16.5%,PCU₄产量最高,分别达13899和12439 kg·hm^-2。因此,不同土层深度配施缓释/普通尿素既能满足玉米生育前期对氮素的需求,也能提高生育后期10～30 cm土层土壤无机氮含量和酶活性,促进玉米生长,增加玉米产量,其中PCU₄处理施肥方式最佳。     

Effects of amiloride and diethyl pyrocarbonate on CSF HCO-3 and ventilation in hypercapnia

Amiloride (10(-3) M), a Na+-H+ countertransport inhibitor, infused into the cisterna magna (10 microliter/min for 40 min) of ketamine-xylazine-anesthetized rabbits decreased the cerebrospinal fluid (CSF) HCO3- response to 3 h of hypercapnia [arterial PCO2 (PaCO2) = 60 Torr] by 21.6% (mean delta CSF [HCO3-]/delta PaCO2 0.232 vs. 0.296 mmol.l-1.Torr-1, P less than 0.05). Diethyl pyrocarbonate (DEPC, 10(-3) M), a histidine-blocking agent, infused into the cisterna magna decreased the CSF HCO3- response to hypercapnia by 25.3% (mean delta CSF [HCO3-]/delta PaCO2, 0.230 vs. 0.308 mmol.l-1.Torr-1, P less than 0.02). DEPC is known to inhibit the ventilatory response to hypercapnia (E. Nattie. Respir. Physiol. 64: 161-176, 1986) by a direct effect at the ventrolateral medulla (E. Nattie. J. Appl. Physiol. 61: 843-850, 1986). In this study amiloride had no significant effect on the ventilatory response to hypercapnia. The interpretation is that a Na+-H+ countertransport protein, perhaps with a histidine at a key location, is involved in CSF acid-base regulation and that amiloride appears to have no effects on the chemoreception process. DEPC appears to have effects on chemoreception and on CSF acid-base regulation.     

Respiratory depression produced by centrally administered taurine in the cat

The effects of taurine (0.8-64.8 mumol) were studied on respiratory activity following intracisternal (cisterna magna) and intracerebroventricular (lateral ventricle) injections in cats anesthetized with alpha-chloralose. Respiratory activity was measured by using a Fleisch pneumotachograph and monitoring tracheal airflow. The flow signal was integrated to obtain tidal volume (VT) and respiratory rate (f) was obtained by counting the number of VT excursions over one minute. Inspiratory (TI), expiratory (TE) and total (TTOT) cycle durations were also determined during this time period. In addition, end-tidal CO2 was continuously monitored. Associated changes in arterial pressure (femoral artery cannula) and heart rate were also determined. After injections into the cisterna magna, taurine caused dose-related decreases in minute ventilation (VE). The maximal decrease in VE was from 495 +/- 59 to 64 +/- 14 ml/min (p less than 0.05), and was due to both decreases in VT (from 27 +/- 3 to 5 +/- 1 ml; p less than 0.05) and f (from 18 +/- 1 to 12 +/- 2 breaths/min; p less than 0.05). TE and TTOT were increased from 2.4 +/- 0.4 to 4.5 +/- 0.6 sec (p less than 0.05) and from 3.7 +/- 0.4 to 6.4 +/- 0.8 sec (p less than 0.05), respectively. Mean inspiratory flow (VT/TI), a measure of inspiratory drive, was decreased from 21 +/- 4 to 4 +/- 2 ml/sec (p less than 0.05). Apnea occurred in 5 of 6 animals after the 64.8 mumol dose. This respiratory depression occurred without any significant change in arterial pressure. After lateral ventricle injections, taurine also caused dose-related, but not as pronounced, decreases in respiratory activity. In addition, taurine caused significant decreases (p less than 0.05) in arterial pressure in doses that decreased VE. Taurine administered intravenously had no significant cardiorespiratory depressant effects. These data indicate that centrally administered taurine produces respiratory depression and, depending on the route of CNS administration, also produces hypotension.     

CT引导下经皮肺穿刺并发症的高危因素分析

目的:探讨CT引导下经皮肤肺穿刺并发症发生的高危因素.方法:回顾分析93例因肺部肿块行CT引导下经皮肤肺穿刺患者,对其性别、吸烟史、病灶大小、肿块类型、组织学分型、穿刺时体位、进针位置、进针角度和进针深度等因素与穿刺后并发症的相关性进行分析.结果:单因素分析显示吸烟史、病灶大小、肿块类型和进针深度在有并发症患者和无并发症的患者之间存在显著性差异(P＜0.05),多因素分析显示病灶大小、肿块类型和进针深度在两组患者间存在显著性差异(P＜0.05).结论:病灶大小、肿块类型和进针深度为CT引导下经皮肺穿刺并发症出现的高危因素.     

Behavioral activation by CRF: evidence for the involvement of the ventral forebrain

Rats injected intracerebroventricularly with corticotropin releasing factor (CRF) at the level of the lateral ventricle or cisterna magna showed a dose-dependent increase in locomotor activity. The increase in locomotor activity from injections of CRF into the cisterna magna was blocked by a cold cream plug in the cerebral aqueduct. An identical plug failed to block the increase in locomotor activity produced by CRF injected into the lateral ventricle. Intracerebral injections of CRF produced a site specific increase in locomotor activity with the largest increases observed from CRF injected into the substantia innominata/lateral preoptic area. Results suggest that the locomotor activating effects of CRF may be due to an activation of CRF receptors in the ventral forebrain, a region rich in CRF cell bodies and projections.