• <tr id="yyy80"></tr>
  • <sup id="yyy80"></sup>
  • <tfoot id="yyy80"><noscript id="yyy80"></noscript></tfoot>
  • 99热精品在线国产_美女午夜性视频免费_国产精品国产高清国产av_av欧美777_自拍偷自拍亚洲精品老妇_亚洲熟女精品中文字幕_www日本黄色视频网_国产精品野战在线观看 ?

    A more consistent intraluminal rhesus monkey model of ischemic stroke

    2014-06-01 09:08:56BoZhaoGuoweiShangJianChenXiaokunGengXinYeGuoxunXuJuWangJiashengZhengHongjunLiFauziaAkbaryShengliLiJingLuFengLingXunmingJi

    Bo Zhao, Guowei Shang, Jian Chen, Xiaokun Geng, Xin Ye, Guoxun Xu, Ju Wang, Jiasheng Zheng, Hongjun Li, Fauzia Akbary, Shengli Li, Jing Lu, Feng Ling, Xunming Ji,

    1 Department of Neurosurgery, Xuanwu Hospital, Capital Medical University, Beijing, China

    2 Department of Anesthesiology, Xuanwu Hospital, Capital Medical University, Beijing, China

    3 Department of Laboratory Animal Science, Capital Medical University, Beijing, China

    4 Radiology Department, Beijing Youan Hospital, Capital Medical University, Beijing, China

    5 Wayne State University School of Medicine, Detroit, MI, USA

    6 Center of Stroke, Beijing Institute for Brain Disorders, Beijing, China

    A more consistent intraluminal rhesus monkey model of ischemic stroke

    Bo Zhao1, Guowei Shang1, Jian Chen1, Xiaokun Geng1, Xin Ye2, Guoxun Xu2, Ju Wang3, Jiasheng Zheng4, Hongjun Li4, Fauzia Akbary5, Shengli Li3, Jing Lu3, Feng Ling1, Xunming Ji1,6

    1 Department of Neurosurgery, Xuanwu Hospital, Capital Medical University, Beijing, China

    2 Department of Anesthesiology, Xuanwu Hospital, Capital Medical University, Beijing, China

    3 Department of Laboratory Animal Science, Capital Medical University, Beijing, China

    4 Radiology Department, Beijing Youan Hospital, Capital Medical University, Beijing, China

    5 Wayne State University School of Medicine, Detroit, MI, USA

    6 Center of Stroke, Beijing Institute for Brain Disorders, Beijing, China

    Endovascular surgery is advantageous in experimentally induced ischemic stroke because it causes fewer cranial traumatic lesions than invasive surgery and can closely mimic the pathophysiology in stroke patients. However, the outcomes are highly variable, which limits the accuracy of evaluations of ischemic stroke studies. In this study, eight healthy adult rhesus monkeys were randomized into two groups with four monkeys in each group: middle cerebral artery occlusion at origin segment (M1) and middle cerebral artery occlusion at M2 segment. The blood fl ow in the middle cerebral artery was blocked completely for 2 hours using the endovascular microcoil placement technique (1 mm × 10 cm) (undetachable), to establish a model of cerebral ischemia. The microcoil was withdrawn and the middle cerebral artery blood fl ow was restored. A reversible middle cerebral artery occlusion model was identi fi ed by hematoxylin-eosin staining, digital subtraction angiography, magnetic resonance angiography, magnetic resonance imaging, and neurological evaluation. The results showed that the middle cerebral artery occlusion model was successfully established in eight adult healthy rhesus monkeys, and ischemic lesions were apparent in the brain tissue of rhesus monkeys at 24 hours after occlusion. The rhesus monkeys had symptoms of neurological de fi cits. Compared with the M1 occlusion group, the M2 occlusion group had lower infarction volume and higher neurological scores. These experimental fi ndings indicate that reversible middle cerebral artery occlusion can be produced with the endovascular microcoil technique in rhesus monkeys. The M2 occluded model had less infarction and less neurological impairment, which offers the potential for application in the fi eld of brain injury research.

    nerve regeneration; brain injury; rhesus monkeys; model middle cerebral artery; microcoil; infarction; stroke; interventional therapy; digital subtraction angiography; magnetic resonance image; neuroimaging; neuroregeneration

    Funding: This study was financially supported by grants from the National Key Basic Research Program (973 Program) of China, No. 2011CB707804; Beijing Municipal Science and Technology Project, No. 2121100005312016.

    Zhao B, Shang GW, Chen J, Geng XK, Ye X, Xu GX, Wang J, Zheng JS, Li HJ, Akbary F, Li SL, Lu J, Ling F, Ji XM. A more consistent intraluminal rhesus monkey model of ischemic stroke. Neural Regen Res. 2014;9(23):2087-2094.

    Introduction

    Although ischemic stroke is one of the leading causes of death, disability, and massive socioeconomic loss worldwide (Roger et al., 2011; Cook et al., 2012), its pathophysiology and biomedicine remain unclear. Because the information obtained from human patients is limited, reproducible animal models of focal ischemic infarction are crucial for studying cerebral ischemia (Liu et al., 2012). In contrast to rodent and other non-primate stroke models that have inherent flaws, nonhuman primate models of focal ischemic stroke provide excellent opportunities for understanding the vascular and cellular pathophysiology of cerebral ischemic injury because they resemble what happens in humans (Fukuda and Zoppo, 2003). In addition to being used for neuroprotective assessment, these models can also facilitate efforts to develop diagnostic tools for identifying and treating stroke symptoms.

    Stroke in humans is highly heterogeneous both clinically and radiographically, and to date, no consensus has been reached regarding which nonhuman stroke models most closely mimics human pathology (Kaku et al., 1998; Liu et al., 2012). The use of a gyrencephalic species such as a rhesus monkey (West et al., 2009) is desirable for studies of stroke in nonhuman primates. Indeed, the rhesus monkey is ideal for stroke studies because its brain is structurally and functionally similar to that of humans (D’Arceuil et al., 2006; Hofer et al., 2008; Kumar et al., 2009; Rodriguez-Mercado et al., 2012). Additionally, the immunologic pro fi le of rhesus monkeys is similar to that of humans (Sariol et al., 2007; Valentine andWatkins, 2008). These characteristics make the rhesus monkey the preferred model for ischemic stroke studies.

    One dif fi culty with stroke models is interanimal variability. Until recently, this issue was incompletely addressed by nonhuman primate models of middle cerebral artery occlusion (MCAO), which were plagued by considerable interanimal variability in terms of infarct size. Several experiments were designed to reduce this variability using an open approach surgical technique (Liu et al., 1992; Huang et al., 2000). Endovascular surgery to induce ischemic stroke has been shown to produce fewer traumatic cranial incisions and to more closely model the pathophysiology in humans compared with invasive surgery. However, the variability in stroke volume was high (D’Arceuil et al, 2006). Using the intraluminal microcoil approach, Rink et al. (2008) reported a 15% standard deviation in infarct volume in canines subjected to transient MCAO, which is significantly better than other methods. However, this method has not been developed in nonhuman primates such as rhesus monkeys. Therefore, in an effort to address stroke variability in primate models, we used an intraluminal approach to modify a model of reperfused stroke, and create a unilateral MCAO. By taking advantage of the microcoil approach, we hypothesized that more-consistent infarctions would occur, thereby allowing experiments involving fewer animals to achieve statistically signi fi cant results. Among heterogeneous stroke subtypes, the proportions of large-arterial and cortical ischemic strokes were reported as 71% and 22% in humans, respectively (Feigin et al., 2006; Halkes et al., 2006). To simulate these, we established and evaluated two types of ischemic stroke models by obstructing different locations in the middle cerebral artery.

    Using an intraluminal technique, we introduced a microcoil into the middle cerebral artery and temporally occluded and then recanalized it. The reliability of infarcts produced by this procedure could be measured by taking magnetic resonance images (MRI) of the monkey brains during the acute phase. The procedure resulted in discrete and limited neurobehavioral deficits. This paper: (1) describes how we created the occluded-stroke model in rhesus monkeys using intraluminal microcoils to generate segmental MCAO; (2) identi fi es and compares the difference in stroke volume and neurological evaluation scores between ischemic lesions induced at different locations in the middle cerebral artery.

    Materials and Methods

    Animals

    A total of eight adult healthy male rhesus monkeys (Macaca mulatta), with an average age of 8.2 ± 1.2 years and an average body weight of 9.40 ± 0.99 kg, were selected for the study (Academy of Military Medical Sciences, China; animal license No. 2013-26). Animals were housed individually indoors under a 12-hour light/dark cycle (light on from 07:00 to 19:00), and at a constant room temperature of 22—24°C. Laboratory diet was provided twice daily, supplemented with fresh fruit and vegetables and drinking water ad libitum. At 1 month before surgery, animals were screened for general health, epidemic diseases, and neurological disorders. All animal experiments were approved and monitored by the Department of Laboratory Animal Science at the Capital Medical University in China. Every effort was made to ensure that the animals were free from pain and discomfort. The principal investigator and the primate handling staff were present for all procedures.

    Anesthesia and monitoring

    All animals were deprived of food for 12 hours before the experiment. Anesthesia was induced with intramuscularly administered ketamine (10 mg/kg; Fujian Gutian Pharmaceutical Co., Fujian Province, China) (Woods et al., 2013). Thereafter, monkeys were positioned in a supine position on a plastic stretcher. The trachea was intubated and ventilated. Both the inguinal regions and anterior pectoral region were shaved free of hair and cleaned with betadine. Animals were anaesthetized with propofol 0.5 mg/kg per hour (Astra Zeneca, Caponago, Italy) continuously throughout the operation and MRI scan. Physical parameters including blood pressure by arm cuff, respiratory frequency, O2saturation, electrocardiogram, and rectal temperature were continuously monitored. Temperature was maintained at 37.5 ± 0.5°C using a heating blanket. Atropine (0.6 mg/kg) was administered intramuscularly before operation. A femoral arterial line was used to monitor blood pressure and blood gases.

    MRI scan

    We monitored the early changes that followed the experimentally-induced stroke (see below) using MRI. Because infarction volume measured from T2/Flair images was shown to be highly correlated with histological estimates (Huang et al., 2000; Gauvrit et al., 2006; Rink et al., 2008; Neumann et al., 2009; West et al., 2009; Schwartz and Pile-Spellman, 2011), evaluation of the infarct lesion was accomplished based on T2 images. All images were performed on a Magnetom Trio MRI Scanner (3.0T; Siemens AG, Siemens Medical Solutions, Erlangen, Germany). Animals were scanned in the supine position 30 minutes before surgery to collect baseline data in the supine position. During the scans, animals were ventilated with an MRI compatible respirator (MRI2550, Surgivet Corp., USA) and monitored with a multi-parameter monitor device (0500-113, Surgivet Corp) under anesthesia. The MRI scans included T2-weighted (high resolution) and magnetic resonance angiography (MRA) scans. The imaging protocols consisted of an axial turbo spin echo T2-weighted scan (repetition time = 3,000, echo time = 300 ms, matrix = 384 × 384, voxel size = 0.4 mm × 0.4 mm × 1.0 mm, slice thickness = 1 mm), and a 3D time of fl ight (repetition time = 22, echo time = 4.92 ms, matrix = 256 × 256, voxel size = 0.5 mm × 0.5 mm × 0.5 mm, slice thickness = 0.5 mm). The MRA scans were repeated at the end of the operation to identify the MCAO. The T2 scans were repeated 24 hours after MCAO was induced and the ischemic volume of the stroke region was recorded using the same parameters. Monitoring and ventilation were continued throughout the procedure. USVIEW V1.1 (USCUBE, Beijing, China) was used to calculate infarct volume from axial T2-weighted MR images by two independent observers.

    Table 2 Life indicators of the rhesus monkeys subjected to middle cerebral artery occlusion

    Table 1 Scoring system for neurological evaluations

    Surgical procedure

    To minimize variations, all surgical procedures were conducted by a single surgeon. At the beginning of the operation, the animal was given a 1,000 U heparin intravenous bolus, followed by an intravenous infusion of 500 U/h. An 18 gage needle was used to puncture the femoral artery percutaneously using the Seldinger technique. A sheath was then placed in the femoral artery using a catheter-introducer kit (Radifocus Introducer, Terumo, Tokyo, Japan). Next, a guide wire (Radifocus Guide Wire M, Terumo) was inserted through the sheath and the tip of the guide wire was forced from the femoral artery into the abdominal aorta, until it reached the ascending aorta as viewed under X-ray fl uoroscopy (images were digitally acquired at a rate of six frames per second). Then it was further forced from the ascending aorta to the right internal carotid artery. While monitoring with X-ray fluoroscopy a 5F-Guiding Catheter (Cordis, Miami Lakes, FL, USA) was inserted along the guide wire and the tip of the catheter was placed at an inlet of the internal carotid artery. After con fi rming that the tip of the catheter was placed in the desired position, the guide wirewas withdrawn. Subsequently, a Prowler 10 microcatheter (Cordis) was inserted into the guiding catheter and the tip of the catheter was pushed forward through the internal carotid artery to an extent closest to the M1 segment. At this stage, a small amount of a contrast agent was slowly injected with a syringe and the fl ow of the contrast agent from the tip of the microcatheter into the middle cerebral artery was confi rmed. A custom microcoil (1 mm × 10 cm, platinum alloy; Achieva Medical Co., Shanghai, China) was then deployed in the M1 segment of the middle cerebral artery (n = 4), or the distal part of the upper trunk of the M2 segment (n = 4) to occlude the artery for 2 hours. The catheter system was secured in position and the animals were transported to the MR department. Physiological monitoring and support were maintained throughout the operation. After removal of the microcoil, a second angiogram con fi rmed that blood fl ow in the middle cerebral artery was restored.

    Figure 1 Images of the M1 segment in the rhesus monkey models of middle cerebral artery occlusion.

    Figure 4s Correlation analysis of infarct volume and neurological evaluation scores in the two groups.

    Figure 2 Images of the M2 segment in the rhesus monkey models of middle cerebral artery occlusion.

    Post-operative management

    After the operation, the interventional devices were withdrawn, the puncture points were compressed for 30 minutes, and then pressure dressed with sterile bandages. After the animals were occluded, postoperative veterinary care was provided to the monkeys for 24 hours. After MRA scanning, animals were placed in a supine position on a padded mattress with the head elevated 30°. In case of pain, an injection of buprenorphine (Buprecare, 20 μg/kg, intramuscular injection; Animalcare, Dunnington, York, UK) was given as deemed necessary (Barnes, 2012).

    Neurological assessment

    Before the T2-image scanning was performed and 24 hours after MCAO was induced, each monkey was assessed with a commonly used non-human primate neurological evaluation scale (Table 1) by two independent observers who were experienced in evaluating neurological de fi cits (Spetzler et al., 1980; Frazee et al., 1998; Mack et al., 2003; D’Arceuil et al., 2006; Rodriguez-Mercado et al., 2012). This scale is weighted heavily on motor function, but also takes into account behavior changes (mental status) and ocular and cranial nerve impairment. Lower scores represent worse functional outcomes.

    Slice preparation and histological detection

    Figure 3 Comparison of infarction volume (A) and neurological de fi cit scores (B) measured after ischemia in the M1 and M2 occlusion groups.

    After MRI scanning was performed, all animals were euthanized and the brain was harvested for histological study. Animals were deeply anaesthetized with 25 mg/kg pentobarbital and perfused transcardially with physiological saline (pH 6.4) containing heparin (50 U/mL). The brain tissue was harvested immediately post-mortem in all eight animals. Aside from cerebral ischemia, none of the animals had evidence of gross pathology (i.e., intraparenchymal hemorrhage, infection, subdural or epidural hematoma). Continuous 5-mm thick coronal sections were collected from each monkey using a brain matrix for rhesus macaques. These coronal sections were immersed in 10% neutral-buffered formalin/PBS for at least 2 days. Each section was embedded in paraf fi n, cut into adjacent slices (3—4 μm thickness), and stained with hematoxylin-eosin. Gross pathological study of the brain sections revealed well-de fi ned cerebral infarctions in all animals on the side of vessel occlusion.

    Statistical analysis

    All data are expressed as the mean ± SD. Statistical analyses were performed using SPSS 22.0 software (IBM Corp, Armonk, NY, USA). Infarction volume and neurological evaluation were analyzed by individual Student’s t-test. The correlation among ischemic damage measured by MRI, and the neurological evaluation (stroke scores) was analyzed using a Pearson parametric correlation. A P-value < 0.05 was considered signi fi cant.

    Results

    Changes of life indicators in the rhesus monkey model of ischemic stroke

    The anesthesia and surgical procedures were well tolerated throughout the procedure by all rhesus monkeys. No parameters were affected by the MCAO in this set of experiments. Physiological parameters remained within the normal range during the experiments (Table 2). All rhesus monkeys recovered from anesthesia and could be evaluated 24 hours after the occlusion was induced.

    Changes of neurobehavioral function in the rhesus monkey model of ischemic stroke

    Ischemia was induced without intraoperative complications in part of the middle cerebral artery in each rhesus monkey using an intraluminal approach for vessel occlusion. Obvious neurological deficits were observed in all rhesus monkeys. MCAO and reperfusion in rhesus monkeys produced a characteristic de fi cit similar to that in human beings with stroke: Awaking from anesthesia, all rhesus monkeys in the M1 occlusion group exhibited severe neurological impairments, such as depression, decreased activity, lethargy, marked hemiplegia of contralateral limbs, decreased muscle tone, and salivation and drooping of the affected corner of the mouth. Rhesus monkeys in the M2 occlusion group appeared to have contralateral upper limb weakness and apraxia, without severe changes in consciousness or lower limb hemiplegia or weakness. Neurological scores were evaluated by two independent observers, with outcomes in the M1 and M2 occlusion groups being 21.00 ± 7.19 and 60.00 ± 16.25, respectively (Table 2). Lower scores represent worse functional outcomes. There was a statistically signi fi cant difference between the two groups (P = 0.0046).

    Assessment of a rhesus monkey model of ischemic strokeviahistological staining, MRI, and DSA

    Anatomical MRI scans of the rhesus monkeys revealed normal brain anatomy before ischemia was induced. MCAO and reperfusion were confirmed by digital subtraction angiography (DSA) and MRA (Figure 1A, B;Figure 2A, B). After releasing the microcoil, DSA and MRA revealed that the distal part of the middle cerebral artery was absent, and all rhesus monkeys showed high signal intensity (ischemia) on T2-weighted images of the corresponding region. In the M1 occlusion group, high-intensity areas indicated that infarcted lesions were localized primarily in the basal ganglia, internal capsule, white matter, and cortex (the middle cerebral artery territory) (Figure 1C). In the M2 occlusion group, infarcts were smaller and localized in the subcortical white matter and in the cortex of the area around the Sylvian fi ssure (Figure 2C). The total infarct volume in the M1 and M2 occlusion groups were 16,456.58 ± 7,108.45 mm3(range: 9,823.39—24,505.08) and 2,285.34 ± 658.33 mm3(range: 1,579.40—3,101.76), respectively (Table 2). The M1 occlusion group showed signi fi cantly greater ischemic volume than the M2 group (P = 0.0074).

    Microscopic examination of brain pathology in the sec-tions revealed well-de fi ned cerebral infarctions in all rhesus monkeys on the side of vessel occlusion. Hematoxylin-eosin staining revealed clear delineations between infarcted and normal brain areas (Figure 1D;Figure 2D). These fi ndings were con fi rmed by the MRI results.

    Both the MRI images and histological examination confi rmed that the M1 occlusion group had a larger infarction volume than did the M2 occlusion group, and was associated with signi fi cantly lower neurological evaluation scores. Statistical analysis of infarction volume and neurological evaluation score showed a signi fi cant difference between the two groups (Figure 3A, B). The neurological evaluation scores after embolization were significantly negatively correlated with infarct volume (r = ?0.8649, P = 0.0056) (Figure 4).

    Discussion

    Based on the survival rate and lifespan studies of rhesus monkey colonies, 1 year for a rhesus monkey is equivalent to about 3 to 4 years for a human (Tigges et al., 1988; Bradley et al., 1999; Peters and Kemper, 2012). Adult rhesus monkeys (Macaca mulatta) live 7—14 years (Colman et al., 2009; Mattison et al., 2012). Thus, the rhesus monkeys introduced in our study were all adults. Peters and Kemper (2012) reported that brain structures in these monkeys do not change signi fi cantly with age. Consequently, the adult rhesus monkeys here met the standard requirements that have been accepted by other stroke studies, although age was not explicitly mentioned in these studies (West et al., 2009; Cook et al., 2012). There are three main ways to occlude vessels: surgical methods, endovascular methods, and photothrombotic methods, which are limited in frequency (Cook et al., 2012). With technological advancements, stroke models using non-human primates have been gradually developed from the early-ligated internal carotid-artery model to surgical clipping the middle cerebral artery in transorbital or craniotomy operations, and eventually to interventional access (O’Brien and Waltz, 1973; West et al., 2009; Cook et al., 2012; Gauberti et al., 2012). Throughout the development of these techniques, brain impairments have become smaller and more precise, and closer to the clinical pathophysiological processes of natural stroke. The early efforts that used clipping/ligation had major limitations (traumatic impairment) on pathophysiological and therapeutic manipulations. Because stroke in humans is not necessarily associated with head trauma, the traumatic aspect of invasive surgery might be viewed as a major confounding factor. In contrast, the most important advantage of the endovascular technique is that it is less invasive compared with an invasive surgical approach. Furthermore, the endovascular techniques can maintain cranial cavity integrity and the neurological function defects can be assessed more precisely and comprehensively. Earlier intravascular stroke models in non-human primates have used glue/thrombus injection or inflatable balloon occluders, which often resulted in variations in occlusion location and extent. Furthermore, thicker intraluminal devices such as balloons and microcatheters were unable to reach the upper trunk of the M2 segment. Even thinner intraluminal devices such as microwire or suture have significant limitations that have been extensively documented in the literature and contribute to the high degree of variability in lesion volume and location (Rink et al., 2008). Studies of the intraluminal thread model have reported that the leftover fi lament in the internal cerebral artery occluded the anterior choroid artery and the hypothalamic artery, thereby producing unintentional subcortical lesions (Gerriets et al., 2004). Consequently, to obtain a relatively consistent stroke model, we have developed an occlusion model using intraluminal techniques that introduce a microcoil into the middle cerebral artery. The microcoil can accurately embolize any segment of the middle cerebral artery trunk. We chose to occlude the middle cerebral artery because strokes are a common and devastating condition, and their treatment remains a major unsolved problem in neuro-critical care (Heinsius et al., 1998; Goldstein et al., 2001; Huttner and Schwab, 2009).

    Detailed studies have shown that high clinical and radiographic heterogeneity in stroke patients is primarily determined by the degree of collateral circulation, especially after MCAO (Min et al., 2000; Gerraty et al., 2002; Lee et al., 2009; Shuaib et al., 2011). Because the organization of the vascular systems in the brain is most similar among primates, the variance in collateral circulation between the non-human primates likely explains the differences in lesion severity (considerable variation in both lesion size and functional outcome). One problem with non-human primate stroke models is the occlusion of the collateral artery. Some researchers have made efforts to overcome the effects of bypassed blood supply by blocking the collateral artery (Huang et al., 2000; Cook et al., 2012). This variability is certainly a complicating factor for study design and needs serious consideration (Sasaki et al., 2011). To develop a consistent model of MCAO stroke in the rhesus monkey, we initially performed a pilot study with different positions of reversible middle cerebral artery (single vessel) trunk occlusion. In our study, we hypothesized that infarction can be induced by the proximal MCAO using a microcoil if the ori fi ces of perforating lenticulostriate arteries were involved, which have well-formed distal cerebral collaterals. As shown byFigure 1, occlusion of the M1 segment blocked the blood fl ow to the whole middle cerebral artery territory including the cortex, subcortical structures, and basal ganglia region. This was attributed to that the occlusion was not a point but a segment of vessel, blocked the opening of collateral arteries branched off to serve these areas. Thus creating a large and consistently sized cerebral infarction may simulate the major hemispheric infarction in humans. Freret et al. (Gao et al., 2006; Liu et al., 2007; Freret et al., 2008) reported a 58% standard deviation around the mean infarct volume from marmosets (n = 4) subjected to transient MCAO, other endovascular models in nonhuman primates have reported larger standard deviations in infarct volume across animals. In our study, when the microcoil approach was used, the standard deviation of the M1 occlusion group was 43% (n = 4), which was less than the aforementioned results. While the standard deviation of the M2 occlusion group was up to 29% (n = 4). The appreciably tighter standard deviation in stroke-induced lesion volumes with the microcoil method suggested that fewer experimental animals would be required to achieve statistical signi fi cance in stroke studies. Nevertheless,owing to the presence of leptomeningeal anastomoses and communication between the extracranial and intracranial circulations, the heterogeneous stroke variation could not be eliminated entirely. The results of the present investigation con fi rmed our hypothesis that the microcoil method could establish a more homogeneous ischemic stroke model in rhesus monkeys.

    Another novelty of this study was that introducing the microcoil into the middle cerebral artery at different positions could establish two types of reversible stroke models. Each model would mimic a subclass of thrombotic stroke in humans and offer a relatively homogenous therapeutic target for studies of neuroprotection. While the M1 occlusion group had more severe neurological outcomes than the M2 occlusion group, the M1 occlusion model successfully simulated malignant middle cerebral artery infarction, while the M2 occlusion model successfully simulated cortical infarction. To the best of our knowledge, our study was the first to establish a single forelimb apraxia model using an intraluminal method, which made it particularly suitable for studies of primate motor function. The occlusion at the upper trunk of the middle cerebral artery M2 segment yielded a ischemic infarct area around the Sylvian fi ssure, including the upper part of Brodmann 4 and 6 areas, which correspond to the distal forelimb in the cortex of primates, thus affecting the hand contralateral to the infarction (Paxinos et al., 2000; Plautz et al., 2003). These results show that the M2 occlusion model may provide a platform for single motor function research. They also confirm the anatomical and species similarity between humans and rhesus monkeys. Although the microcoil has been applied in other non-primate animal stroke models (Rink et al., 2008), they did not take advantage of its superior qualities. Non-human primates have apparent functional differences in fi ne dexterous ability of the forelimbs, which makes them more suitable for microcoil application. These differences enabled more accurate measurement by neurological assessment of impairments and therapeutic outcome of the Brodmann 4 and 6 in rhesus monkeys. Furthermore, because of the significant correlations between stroke volume and neurological de fi ciency, the M2 occlusion models had milder neurological deficits and better self-care abilities, which inevitably decreased the expense and effort needed in postoperative care. Because motor function could be assessed precisely while maintaining the aforementioned homogeneity in ischemic volume, the M2-segmental MCAO model provided a relative standard tool for ischemic stroke studies and therapeutic evaluation.

    Imaging fi ndings and functional assessment in this study demonstrated that an intraluminally-released microcoil could establish versatile reversible MCAO in rhesus monkey models. These modi fi ed intravascular stroke models in rhesus monkeys would mimic two types of ischemic stroke in humans and offer a relatively homogenous therapeutic target for the clinical and fundamental study of ischemic stroke. Comparing M1-occluded and M2-occluded models in rhesus monkeys, brain damage was signi fi cantly greater in the M1-occluded models than that in M2-occluded models, and the M2-occluded MCAO model was less variable. The resulting neurological de fi cits correlated well with ischemic stroke volume.

    Author contributions:Zhao B and Ji XM designed the study. Zhao B, Shang GW, Chen J, Geng XK, Ye X, Xu GX, Wang J, Zheng JS, Li HJ, Li SL, Lu J, Ling F and Ji XM performed the experiments. Zhao B analyzed data. Zhao B and Akbary F wrote the paper. All authors approved the final version of the manuscript.

    Con fl icts of interest:None declared.

    Barnes DC (2012) Subtotal colectomy by rectal pull-through for treatment of idiopathic megacolon in 2 cats. Can Vet J 53:780-782.

    Bradley DV, Fernandes A, Lynn M, Tigges M, Boothe RG (1999) Emmetropization in the rhesus monkey (Macaca mulatta): birth to young adulthood. Invest Ophthalmol Vis Sci 40:214-229.

    Colman RJ, Anderson RM, Johnson SC, Kastman EK, Kosmatka KJ, Beasley TM, Allison DB, Cruzen C, Simmons HA, Kemnitz JW, Weindruch R (2009) Caloric restriction delays disease onset and mortality in rhesus monkeys. Science 325:201-204.

    Cook DJ, Tymianski M (2012) Nonhuman primate models of stroke for translational neuroprotection research. Neurotherapeutics 9:371-379.

    Cook DJ, Teves L, Tymianski M (2012) Treatment of stroke with a PSD-95 inhibitor in the gyrencephalic primate brain. Nature 483:213-217.

    D’Arceuil HE, Duggan M, He J, Pryor J, de Crespigny A (2006) Middle cerebral artery occlusion in Macaca fascicularis: acute and chronic stroke evolution. J Med Primatol 35:78-86.

    Feigin V, Carter K, Hackett M, Barber PA, McNaughton H, Dyall L, Chen MH, Anderson C (2006) Ethnic disparities in incidence of stroke subtypes: auckland regional community stroke study, 2002-2003. Lancet Neurol 5:130-139.

    Frazee JG, Luo X, Luan G, Hinton DS, Hovda DA, Shiroishi MS, Barcliff LT, Kontos HA (1998) Retrograde transvenous neuroperfusion: a back door treatment for stroke editorial comment. Stroke 29:1912-1916.

    Freret T, Bouet V, Toutain J, Saulnier R, Pro-Sistiaga P, Bihel E, Mackenzie ET, Roussel S, Schumann-Bard P, Touzani O (2008) Intraluminal thread model of focal stroke in the non-human primate. J Cereb Blood Flow Metab 28:786-796.

    Fukuda S, Zoppo GJd (2003) Models of focal cerebral ischemia in the nonhuman primate. ILAR J 44:96-104.

    Gao H, Liu Y, Lu S, Xiang B, Wang C (2006) A reversible middle cerebral artery occlusion model using intraluminal balloon technique in monkeys. J Stroke Cerebrovasc Dis 15:202-208.

    Garcia JH (1984) Experimental ischemic stroke: a review. Stroke 15:5-14.

    Gauberti M, Obiang P, Guedin P, Balossier A, Gakuba C, Diependaele AS, Chazalviel L, Vivien D, Young AR, Agin V, Orset C (2012) Thrombotic stroke in the anesthetized monkey (Macaca mulatta): characterization by MRI-a pilot study. Cerebrovasc Dis 33:329-339.

    Gauvrit JY, Leclerc X, Girot M, Cordonnier C, Sotoares G, Henon H, Pertuzon B, Michelin E, Devos D, Pruvo JP, Leys D (2006) Fluid-attenuated inversion recovery (FLAIR) sequences for the assessment of acute stroke: inter observer and inter technique reproducibility. J Neurol 253:631-635.

    Gerraty RP, Parsons MW, Barber PA, Darby DG, Desmond PM, Tress BM, Davis SM (2002) Examining the lacunar hypothesis with diffusion and perfusion magnetic resonance imaging. Stroke 33:2019-2024.

    Gerriets T, Stolz E, Walberer M, Muller C, Rottger C, Kluge A, Kaps M, Fisher M, Bachmann G (2004) Complications and pitfalls in rat stroke models for middle cerebral artery occlusion: a comparison between the suture and the macrosphere model using magnetic resonance angiography. Stroke 35:2372-2377.

    Ginsberg MD (2002) Adventures in the pathophysiology of brain ischemia: penumbra, gene expression, neuroprotection: the 2002 thomas willis lecture. Stroke 34:214-223.

    Goldstein LB, Adams R, Becker K, Furberg CD, Gorelick PB, Hademenos G, Hill M, Howard G, Howard VJ, Jacobs B, Levine SR, Mosca L, Sacco RL, Sherman DG, Wolf PA, del Zoppo GJ (2001) Primary prevention of ischemic stroke : a statement for healthcare professionals from the stroke council of the american heart association. Stroke 32:280-299.

    Green AR (2002) Why do neuroprotective drugs that are so promising in animals fail in the clinic? An industry perspective. Clin Exp Pharmacol Physiol 29:1030-1034.

    Halkes PH, Kappelle LJ, van Gijn J, van Wijk I, Koudstaal PJ, Algra A (2006) Large subcortical infarcts: clinical features, risk factors, and long-term prognosis compared with cortical and small deep infarcts. Stroke 37:1828-1832.

    Heinsius T, Bogousslavsky J, Van Melle G (1998) Large infarcts in the middle cerebral artery territory Etiology and outcome patterns. Neurology 50:341-350.

    Hofer S, Merboldt KD, Tammer R, Frahm J (2008) Rhesus monkey and human share a similar topography of the corpus callosum as revealed by diffusion tensor MRI in vivo. Cerebral cortex 18:1079-1084.

    Huang J, Mocco J, Choudhri TF, Poisik A, Popilskis SJ, Emerson R, DelaPaz RL, Khandji AG, Pinsky DJ, Connolly ES, Zlokovic BV (2000) A modi fi ed transorbital baboon model of reperfused stroke editorial comment. Stroke 31:3054-3063.

    Huttner HB, Schwab S (2009) Malignant middle cerebral artery infarction: clinical characteristics, treatment strategies, and future perspectives. Lancet Neurol 8:949-958.

    Kaku S, Umemura K, Mizuno A, Yano S, Suzuki Ki, Kawasaki T, Nakashima M (1998) Evaluation of a GPIIb/IIIa antagonist YM337 in a primate model of middle cerebral artery thrombosis. Eur J Pharmacol 345:185-192.

    Kumar N, Lee JJ, Perlmutter JS, Derdeyn CP (2009) Cervical carotid and circle of willis arterial anatomy of macaque monkeys: a comparative anatomy study. Anat Rec (Hoboken) 292:976-984.

    Kwiecien TD, Sy C, Ding Y (2014) Rodent models of ischemic stroke lack translational relevance. are baboon models the answer? Neurol Res 36:417-422.

    Lee KY, Latour LL, Luby M, Hsia AW, Merino JG, Warach S (2009) Distal hyperintense vessels on FLAIR: an MRI marker for collateral circulation in acute stroke? Neurology 72:1134-1139.

    Liu S, Hu WX, Zu QQ, Lu SS, Xu XQ, Sun L, Zhou WZ, Shi HB (2012) A novel embolic stroke model resembling lacunar infarction following proximal middle cerebral artery occlusion in beagle dogs. J Neurosci Methods 209:90-96.

    Liu XG, Branston NM, Kawauchi M, Symon L (1992) A model of acute focal ischemia in the territory of the anterior cerebral artery in baboons. Stroke 23:40-44.

    Liu Y, D’Arceuil HE, Westmoreland S, He J, Duggan M, Gonzalez RG, Pryor J, de Crespigny AJ (2007) Serial diffusion tensor MRI after transient and permanent cerebral ischemia in nonhuman primates. Stroke 38:138-145.

    Mack WJ, Komotar RJ, Mocco J, Coon AL, Hoh DJ, King RG, Ducruet AF, Ransom ER, Oppermann M, DeLaPaz R, Connolly ES Jr. (2003) Serial magnetic resonance imaging in experimental primate stroke: validation of MRI for pre-clinical cerebroprotective trials. Neurol Res 25:846-852.

    Marshall JW, Duf fi n KJ, Green AR, Ridley RM, Finklestein SP (2001) NXY-059, a free radical-trapping agent, substantially lessens the functional disability resulting from cerebral ischemia in a primate species editorial comment. Stroke 32:190-198.

    Marshall JW, Cummings RM, Bowes LJ, Ridley RM, Green AR (2003a) Functional and histological evidence for the protective effect of NXY-059 in a primate model of stroke when given 4 hours after occlusion. Stroke 34:2228-2233.

    Marshall JW, Green AR, Ridley RM (2003b) Comparison of the neuroprotective effect of clomethiazole, AR-R15896AR and NXY-059 in a primate model of stroke using histological and behavioural measures. Brain Res 972:119-126.

    Mattison JA, Roth GS, Beasley TM, Tilmont EM, Handy AM, Herbert RL, Longo DL, Allison DB, Young JE, Bryant M, Barnard D, Ward WF, Qi W, Ingram DK, de Cabo R (2012) Impact of caloric restriction on health and survival in rhesus monkeys from the NIA study. Nature 489:318-321.

    Min WK, Park KK, Kim YS, Park HC, Kim JY, Park SP, Suh CK (2000) Atherothrombotic middle cerebral artery territory infarction: topographic diversity with common occurrence of concomitant small cortical and subcortical infarcts. Stroke 31:2055-2061.

    Neumann AB, Jonsdottir KY, Mouridsen K, Hjort N, Gyldensted C, Bizzi A, Fiehler J, Gasparotti R, Gillard JH, Hermier M, Kucinski T, Larsson EM, Sorensen L, Ostergaard L (2009) Interrater agreement for fi nal infarct MRI lesion delineation. Stroke 40:3768-3771.

    O’Brien MD, Waltz AG (1973) Transorbital approach for occluding the middle cerebral artery without craniectomy. Stroke 4:201-206.

    Paxinos G, Huang XF, Toga AW (2000) The Rhesus Monkey Brain in Stereotaxic Coordinates. Sat Lake City, Academic Press, USA.

    Peters A, Kemper T (2012) A review of the structural alterations in the cerebral hemispheres of the aging rhesus monkey. Neurobiol Aging 33:2357-2372.

    Plautz EJ, Barbay S, Frost SB, Friel KM, Dancause N, Zoubina EV, Stowe AM, Quaney BM, Nudo RJ (2003) Post-infarct cortical plasticity and behavioral recovery using concurrent cortical stimulation and rehabilitative training: a feasibility study in primates. Neurol Res 25:801-810.

    Rink C, Christoforidis G, Abduljalil A, Kontzialis M, Bergdall V, Roy S, Khanna S, Slivka A, Knopp M, Sen CK (2008) Minimally invasive neuroradiologic model of preclinical transient middle cerebral artery occlusion in canines. Proc Natl Acad Sci U S A 105:14100-14105.

    Rodriguez-Mercado R, Ford GD, Xu Z, Kraiselburd EN, Martinez MI, Eterovi? VA, Colon E, Rodriguez IV, Portilla P, Ferchmin PA, Gierbolini L, Rodriguez-Carrasquillo M, Powell MD, Pulliam JV, McCraw CO, Gates A, Ford BD (2012) Acute neuronal injury and blood genomic pro fi les in a nonhuman primate model for ischemic stroke. Comp Med 62:427-438.

    Roger VL, Go AS, Lloyd-Jones DM, Adams RJ, Berry JD, Brown TM, Carnethon MR, Dai S, de Simone G, Ford ES, Fox CS, Fullerton HJ, Gillespie C, Greenlund KJ, Hailpern SM, Heit JA, Ho PM, Howard VJ, Kissela BM, Kittner SJ, et al (2011) Heart disease and stroke statistics--2011 update: a report from the American Heart Association. Circulation 123:e18-e209.

    Sariol CA, Munoz-Jordan JL, Abel K, Rosado LC, Pantoja P, Giavedoni L, Rodriguez IV, White LJ, Martinez M, Arana T, Kraiselburd EN (2007) Transcriptional activation of interferon-stimulated genes but not of cytokine genes after primary infection of rhesus macaques with dengue virus type 1. Clin Vaccine Immunol 14:756-766.

    Sasaki M, Honmou O, Radtke C, Kocsis JD (2011) Development of a middle cerebral artery occlusion model in the nonhuman primate and a safety study of i.v. infusion of human mesenchymal stem cells. PLoS One 6:e26577.

    Saver JL, Albers GW, Dunn B, Johnston KC, Fisher M, Consortium SV (2009) stroke therapy academic industry roundtable (STAIR) recommendations for extended window acute stroke therapy trials. Stroke 40:2594-2600.

    Sawada K, Fukunishi K, Kashima M, Imai N, Saito S, Sakata-Haga H, Aoki I, Fukui Y (2012) Neuroanatomic and magnetic resonance imaging references for normal development of cerebral sulci of laboratory primate, cynomolgus monkeys (Macaca fascicularis). Congenit Anom (Kyoto) 52:16-27.

    Schwartz AE, Pile-Spellman J (2011) New model of reperfused stroke by occlusion of the anterior cerebral artery in baboons. Acta Neurochirurgica 153:327-331.

    Shuaib A, Butcher K, Mohammad AA, Saqqur M, Liebeskind DS (2011) Collateral blood vessels in acute ischaemic stroke: a potential therapeutic target. Lancet Neurol 10:909-921.

    Shuaib A, Lees KR, Lyden P, Grotta J, Davalos A, Davis SM, Diener HC, Ashwood T, Wasiewski WW, Emeribe U, Investigators SIT (2007) NXY-059 for the treatment of acute ischemic stroke. New Engl J Med 357:562-571.

    Spetzler R, Selman W, Weinstein P, Townsend J, Mehdorn M, Telles D, Crumrine R, Macko R (1980) Chronic reversible cerebral ischemia evaluation of a new baboon model. Neurosurgery 7:5.

    Tigges J, Gordon TP, McClure HM, Hall EC, Peters A (1988) Survival rate and life span of rhesus monkeys at the Yerkes regional primate research center. Am J Primatol 15:263-273.

    Valentine LE, Watkins DI (2008) Relevance of studying T cell responses in SIV-infected rhesus macaques. Trends Microbiol 16:605-611.

    West GA, Golshani KJ, Doyle KP, Lessov NS, Hobbs TR, Kohama SG, Pike MM, Kroenke CD, Grafe MR, Spector MD, Tobar ET, Simon RP, Stenzel-Poore MP (2009) A new model of cortical stroke in the rhesus macaque. J Cereb Blood Flow Metab 29:1175-1186.

    Woods SE, Marini RP, Patterson MM (2013) Noninvasive temporal artery thermometry as an alternative to rectal thermometry in research macaques (Macaca spp.). J Am Assoc Lab Anim Sci 52:295-300.

    Copyedited by Norman C, Yang Y, Li CH, Song LP, Wang J, Zhao M

    10.4103/1673-5374.147936

    Xunming Ji, Department of Neurology, Xuanwu Hospital, Capital Medical University, Center of Stroke, Beijing Institute for Brain Disorders, Beijing, China 100053, Beijing, China, jixm@ccmu.edu.cn.

    http://www.nrronline.org/

    Accepted: 2014-11-22

    色综合站精品国产| 日韩欧美一区视频在线观看 | 深爱激情五月婷婷| 国产爱豆传媒在线观看| 午夜视频国产福利| 国产单亲对白刺激| 午夜福利在线观看吧| 久久久精品94久久精品| 午夜福利高清视频| 免费观看在线日韩| 久久久久国产网址| 尤物成人国产欧美一区二区三区| 国产高清有码在线观看视频| 有码 亚洲区| av播播在线观看一区| 不卡视频在线观看欧美| 高清在线视频一区二区三区| 国产精品嫩草影院av在线观看| 欧美精品国产亚洲| 日韩精品青青久久久久久| 国产精品1区2区在线观看.| 十八禁国产超污无遮挡网站| 18禁动态无遮挡网站| 成人av在线播放网站| 嫩草影院入口| 插阴视频在线观看视频| 亚洲18禁久久av| 日本与韩国留学比较| 三级经典国产精品| 免费人成在线观看视频色| 亚洲精品一区蜜桃| 亚洲国产色片| av在线播放精品| 国产亚洲91精品色在线| 精品午夜福利在线看| 国产精品一区二区三区四区久久| 国产男人的电影天堂91| 在线观看一区二区三区| 日韩精品青青久久久久久| 亚洲色图av天堂| 日韩在线高清观看一区二区三区| 一级爰片在线观看| 啦啦啦啦在线视频资源| 午夜精品在线福利| 日本爱情动作片www.在线观看| 一级片'在线观看视频| 人妻夜夜爽99麻豆av| .国产精品久久| 九九在线视频观看精品| 插逼视频在线观看| av网站免费在线观看视频 | www.av在线官网国产| 男女那种视频在线观看| 久久精品久久久久久久性| 日韩亚洲欧美综合| 毛片一级片免费看久久久久| 一级片'在线观看视频| 热99在线观看视频| av播播在线观看一区| 一区二区三区高清视频在线| 国产成人精品福利久久| 免费看不卡的av| 又爽又黄a免费视频| 国产精品伦人一区二区| 日韩 亚洲 欧美在线| 欧美xxxx黑人xx丫x性爽| 噜噜噜噜噜久久久久久91| 少妇的逼好多水| 日本与韩国留学比较| 国产高清不卡午夜福利| 久久久久久久久中文| 久久精品熟女亚洲av麻豆精品 | 国产乱来视频区| 18禁裸乳无遮挡免费网站照片| 在线观看免费高清a一片| 日韩av在线大香蕉| 日本免费a在线| 中国国产av一级| 欧美精品国产亚洲| 亚洲av免费在线观看| 国产极品天堂在线| av在线蜜桃| 国产精品人妻久久久久久| 国产欧美日韩精品一区二区| 中文字幕免费在线视频6| 亚洲高清免费不卡视频| 嫩草影院入口| 搡老乐熟女国产| 97超视频在线观看视频| 蜜臀久久99精品久久宅男| 国产高清三级在线| 天堂影院成人在线观看| 欧美 日韩 精品 国产| 成人av在线播放网站| videossex国产| 午夜福利视频精品| 亚洲国产最新在线播放| 亚洲av免费高清在线观看| 一二三四中文在线观看免费高清| 综合色丁香网| 成人午夜高清在线视频| 97在线视频观看| 免费观看性生交大片5| 国产黄色小视频在线观看| 国产亚洲精品久久久com| 久久久久性生活片| 麻豆av噜噜一区二区三区| 麻豆精品久久久久久蜜桃| 男人狂女人下面高潮的视频| 晚上一个人看的免费电影| 国产欧美日韩精品一区二区| 熟女电影av网| 久久这里只有精品中国| 女人十人毛片免费观看3o分钟| 国产精品美女特级片免费视频播放器| 成人性生交大片免费视频hd| 国产成人aa在线观看| 婷婷色av中文字幕| 国产精品伦人一区二区| 免费看日本二区| 国产69精品久久久久777片| 三级毛片av免费| 国产黄色视频一区二区在线观看| 成人二区视频| 蜜桃久久精品国产亚洲av| 免费少妇av软件| 久久久久免费精品人妻一区二区| 久久亚洲国产成人精品v| 国产精品一区二区三区四区免费观看| 国产永久视频网站| 国产黄片美女视频| 亚洲电影在线观看av| 国产精品人妻久久久久久| 亚洲精品456在线播放app| 日本三级黄在线观看| 精品国内亚洲2022精品成人| 伊人久久精品亚洲午夜| 欧美区成人在线视频| 国产真实伦视频高清在线观看| 久久久久久国产a免费观看| 搡老妇女老女人老熟妇| 美女cb高潮喷水在线观看| 人妻系列 视频| 秋霞在线观看毛片| 91精品国产九色| 晚上一个人看的免费电影| 久久久久久久大尺度免费视频| 免费看日本二区| 狠狠精品人妻久久久久久综合| 久久久久久久国产电影| 乱码一卡2卡4卡精品| 日韩欧美国产在线观看| 老师上课跳d突然被开到最大视频| 老师上课跳d突然被开到最大视频| av在线老鸭窝| 成人美女网站在线观看视频| 观看免费一级毛片| 国产日韩欧美在线精品| 国产一区有黄有色的免费视频 | 欧美3d第一页| 精品久久久久久成人av| 亚洲国产精品国产精品| 日韩视频在线欧美| 中文字幕av在线有码专区| 高清视频免费观看一区二区 | 免费电影在线观看免费观看| 免费电影在线观看免费观看| 一边亲一边摸免费视频| 欧美另类一区| 床上黄色一级片| 一夜夜www| 丝袜美腿在线中文| 性插视频无遮挡在线免费观看| 免费观看的影片在线观看| 免费黄色在线免费观看| 日韩一区二区三区影片| 成人亚洲精品av一区二区| 亚洲国产成人一精品久久久| 别揉我奶头 嗯啊视频| 精品亚洲乱码少妇综合久久| 欧美一级a爱片免费观看看| 精品久久久噜噜| 亚洲av中文字字幕乱码综合| 看黄色毛片网站| 欧美激情国产日韩精品一区| 亚洲经典国产精华液单| 看十八女毛片水多多多| 人人妻人人澡人人爽人人夜夜 | 婷婷色综合大香蕉| 日本wwww免费看| 国产一区有黄有色的免费视频 | 秋霞伦理黄片| 欧美变态另类bdsm刘玥| 人妻制服诱惑在线中文字幕| 波野结衣二区三区在线| 大陆偷拍与自拍| 国产精品综合久久久久久久免费| 午夜日本视频在线| 91aial.com中文字幕在线观看| eeuss影院久久| 97超视频在线观看视频| 色哟哟·www| 日韩欧美精品免费久久| 成人鲁丝片一二三区免费| 国产欧美日韩精品一区二区| 久久久久久久久久成人| 三级经典国产精品| 日韩人妻高清精品专区| 亚洲国产精品sss在线观看| 日日撸夜夜添| 久久综合国产亚洲精品| 在线观看免费高清a一片| 精品一区二区三区视频在线| 久久精品久久久久久久性| 99热这里只有精品一区| 乱码一卡2卡4卡精品| a级一级毛片免费在线观看| 久久久久国产网址| 欧美三级亚洲精品| av在线观看视频网站免费| 看非洲黑人一级黄片| 床上黄色一级片| 校园人妻丝袜中文字幕| 国语对白做爰xxxⅹ性视频网站| 亚洲精品第二区| 欧美 日韩 精品 国产| 欧美一级a爱片免费观看看| 国产成人freesex在线| 亚洲欧美精品自产自拍| 久久国产乱子免费精品| 热99在线观看视频| 国产亚洲午夜精品一区二区久久 | 久久久久久久久久黄片| 亚洲欧洲日产国产| 成人午夜高清在线视频| 一级毛片我不卡| 精品欧美国产一区二区三| 欧美3d第一页| 国产欧美日韩精品一区二区| 成人高潮视频无遮挡免费网站| 看非洲黑人一级黄片| 欧美zozozo另类| 在线免费观看不下载黄p国产| 蜜桃久久精品国产亚洲av| 人人妻人人看人人澡| 欧美xxⅹ黑人| 婷婷六月久久综合丁香| 一级毛片久久久久久久久女| 国产亚洲精品久久久com| 啦啦啦啦在线视频资源| 赤兔流量卡办理| 九九在线视频观看精品| 亚洲av成人精品一二三区| 久久精品久久精品一区二区三区| 简卡轻食公司| 久久久a久久爽久久v久久| 一个人看的www免费观看视频| 亚洲av中文av极速乱| 日韩大片免费观看网站| 国产高清有码在线观看视频| 久久人人爽人人片av| 国产色爽女视频免费观看| 女人久久www免费人成看片| 婷婷色麻豆天堂久久| 日本wwww免费看| 一级二级三级毛片免费看| 男人和女人高潮做爰伦理| 干丝袜人妻中文字幕| 国产精品麻豆人妻色哟哟久久 | 久久久久久伊人网av| 亚洲国产欧美人成| 午夜福利视频精品| 99热全是精品| 日日啪夜夜爽| 欧美人与善性xxx| 纵有疾风起免费观看全集完整版 | 免费av毛片视频| or卡值多少钱| 卡戴珊不雅视频在线播放| 精品人妻熟女av久视频| 极品少妇高潮喷水抽搐| 国产伦一二天堂av在线观看| 精品熟女少妇av免费看| 日韩伦理黄色片| 爱豆传媒免费全集在线观看| 日本av手机在线免费观看| 婷婷色综合www| 特大巨黑吊av在线直播| 成人欧美大片| 国产精品1区2区在线观看.| 午夜福利在线在线| 亚洲av福利一区| 嫩草影院入口| 国产亚洲一区二区精品| 久久久a久久爽久久v久久| 久久久久久久久久成人| 22中文网久久字幕| 国产 亚洲一区二区三区 | 色哟哟·www| 波多野结衣巨乳人妻| 十八禁网站网址无遮挡 | 国产有黄有色有爽视频| 自拍偷自拍亚洲精品老妇| 五月玫瑰六月丁香| 国产精品久久久久久av不卡| 99热全是精品| 国产色婷婷99| 精品国产一区二区三区久久久樱花 | 国产亚洲精品久久久com| 欧美成人精品欧美一级黄| 91狼人影院| 亚洲av中文av极速乱| 日韩欧美三级三区| 一区二区三区四区激情视频| 秋霞在线观看毛片| 国产午夜精品一二区理论片| 国产免费福利视频在线观看| 国产精品国产三级国产av玫瑰| 欧美成人一区二区免费高清观看| 晚上一个人看的免费电影| kizo精华| 91久久精品电影网| 亚洲av福利一区| 亚洲欧美清纯卡通| 精品久久久久久久久久久久久| www.av在线官网国产| 久久99蜜桃精品久久| 国国产精品蜜臀av免费| 禁无遮挡网站| 精品人妻一区二区三区麻豆| 黄色配什么色好看| 精品人妻视频免费看| 中国国产av一级| 熟女人妻精品中文字幕| 亚洲人成网站高清观看| av天堂中文字幕网| 九九久久精品国产亚洲av麻豆| 久久99热这里只频精品6学生| 国产亚洲一区二区精品| 午夜免费男女啪啪视频观看| 一区二区三区高清视频在线| 国产精品一区二区三区四区久久| 好男人视频免费观看在线| 欧美日韩亚洲高清精品| 免费大片黄手机在线观看| 中文欧美无线码| 最近视频中文字幕2019在线8| 熟女人妻精品中文字幕| 免费看不卡的av| 精品一区二区免费观看| 精品一区二区三区视频在线| 亚洲人成网站高清观看| ponron亚洲| 日本猛色少妇xxxxx猛交久久| 午夜福利视频1000在线观看| 最近最新中文字幕大全电影3| av天堂中文字幕网| 菩萨蛮人人尽说江南好唐韦庄| 亚洲aⅴ乱码一区二区在线播放| 如何舔出高潮| 国产成人一区二区在线| 久久99蜜桃精品久久| 成人漫画全彩无遮挡| 人妻系列 视频| 免费看不卡的av| 神马国产精品三级电影在线观看| 国产精品日韩av在线免费观看| 国产精品一区www在线观看| av专区在线播放| 免费高清在线观看视频在线观看| 国产精品爽爽va在线观看网站| 久久精品夜色国产| 人人妻人人看人人澡| 男人舔奶头视频| 舔av片在线| 久久99热6这里只有精品| 天堂av国产一区二区熟女人妻| 在线观看av片永久免费下载| 精品久久国产蜜桃| 最近的中文字幕免费完整| 久久久久久久久中文| 久久久午夜欧美精品| 97在线视频观看| 日韩一区二区三区影片| 麻豆成人av视频| 97人妻精品一区二区三区麻豆| 免费观看无遮挡的男女| 国产久久久一区二区三区| 老司机影院毛片| 免费看a级黄色片| 18禁在线播放成人免费| 午夜福利成人在线免费观看| 国产精品.久久久| 97超视频在线观看视频| 国产成人aa在线观看| 内射极品少妇av片p| 久久精品国产自在天天线| 久久午夜福利片| 亚洲国产精品国产精品| 日韩一本色道免费dvd| 少妇猛男粗大的猛烈进出视频 | 国产一级毛片在线| 夫妻性生交免费视频一级片| 一级毛片电影观看| 午夜精品国产一区二区电影 | 国产精品蜜桃在线观看| 国产男女超爽视频在线观看| 久久久国产一区二区| 干丝袜人妻中文字幕| 三级男女做爰猛烈吃奶摸视频| 亚洲av福利一区| 三级国产精品片| 免费无遮挡裸体视频| 一本—道久久a久久精品蜜桃钙片 精品乱码久久久久久99久播 | 日产精品乱码卡一卡2卡三| 性插视频无遮挡在线免费观看| 日本一本二区三区精品| 亚洲精品乱码久久久久久按摩| 欧美性感艳星| 亚洲在线观看片| 亚洲熟女精品中文字幕| 夜夜看夜夜爽夜夜摸| 久久这里只有精品中国| 大香蕉97超碰在线| 日韩成人伦理影院| 国产精品国产三级国产av玫瑰| 国产中年淑女户外野战色| 少妇熟女aⅴ在线视频| 久久久久久久久大av| 日韩av在线大香蕉| 成人二区视频| 日本免费在线观看一区| 免费播放大片免费观看视频在线观看| 午夜福利在线观看吧| 国产精品99久久久久久久久| 精品午夜福利在线看| 干丝袜人妻中文字幕| 大话2 男鬼变身卡| 欧美日韩视频高清一区二区三区二| 国产大屁股一区二区在线视频| 亚洲精品乱码久久久久久按摩| 日韩av在线大香蕉| 中国美白少妇内射xxxbb| 精品久久久久久电影网| 青春草国产在线视频| 亚洲在久久综合| 国产精品人妻久久久影院| 麻豆乱淫一区二区| 国产精品1区2区在线观看.| 夫妻午夜视频| 中文字幕亚洲精品专区| 婷婷色麻豆天堂久久| 亚洲高清免费不卡视频| 精品久久久久久久人妻蜜臀av| 久久精品国产自在天天线| 午夜爱爱视频在线播放| 人妻制服诱惑在线中文字幕| 国产成人freesex在线| 免费观看在线日韩| 有码 亚洲区| 国产在线一区二区三区精| 亚洲av国产av综合av卡| 亚洲婷婷狠狠爱综合网| 小蜜桃在线观看免费完整版高清| 成人国产麻豆网| 亚洲人与动物交配视频| 色网站视频免费| 国产精品久久久久久精品电影| 亚洲人与动物交配视频| h日本视频在线播放| 激情 狠狠 欧美| 色尼玛亚洲综合影院| 免费高清在线观看视频在线观看| 欧美成人a在线观看| 免费av观看视频| 色吧在线观看| 超碰97精品在线观看| 亚洲电影在线观看av| 观看美女的网站| 大又大粗又爽又黄少妇毛片口| 久久精品夜夜夜夜夜久久蜜豆| 国产美女午夜福利| 久久久久国产网址| 国产成人福利小说| 精品亚洲乱码少妇综合久久| 国产三级在线视频| 国产淫片久久久久久久久| 免费无遮挡裸体视频| 舔av片在线| 亚洲在线自拍视频| 日韩不卡一区二区三区视频在线| 成年免费大片在线观看| 国产高潮美女av| videossex国产| 免费看不卡的av| 亚洲精品色激情综合| 久久久久久久国产电影| 人人妻人人澡欧美一区二区| 大香蕉97超碰在线| 亚洲成色77777| 亚洲av一区综合| 久久精品熟女亚洲av麻豆精品 | 亚洲精品色激情综合| 久久草成人影院| 免费看a级黄色片| 久久久久性生活片| 免费看a级黄色片| 国产精品一区www在线观看| 久久久久久久久久人人人人人人| 深爱激情五月婷婷| 大又大粗又爽又黄少妇毛片口| 久久这里只有精品中国| 中文字幕亚洲精品专区| 久久国产乱子免费精品| 水蜜桃什么品种好| 欧美最新免费一区二区三区| 观看美女的网站| 午夜免费观看性视频| 亚洲av.av天堂| 亚洲av电影不卡..在线观看| 街头女战士在线观看网站| 亚洲人与动物交配视频| 日本-黄色视频高清免费观看| 一区二区三区四区激情视频| 国产伦一二天堂av在线观看| 久久久精品94久久精品| 黄色欧美视频在线观看| 亚洲欧洲国产日韩| 男人狂女人下面高潮的视频| 国产黄频视频在线观看| 在现免费观看毛片| 三级经典国产精品| 国精品久久久久久国模美| 人人妻人人看人人澡| 国国产精品蜜臀av免费| a级一级毛片免费在线观看| www.色视频.com| 国产91av在线免费观看| 日韩欧美精品v在线| 国模一区二区三区四区视频| 欧美激情久久久久久爽电影| or卡值多少钱| 免费av观看视频| 国国产精品蜜臀av免费| 天堂网av新在线| 亚洲国产av新网站| 国产精品伦人一区二区| 亚洲成人精品中文字幕电影| 99热6这里只有精品| 亚洲欧美一区二区三区黑人 | 日日摸夜夜添夜夜添av毛片| 国产亚洲午夜精品一区二区久久 | 精品人妻一区二区三区麻豆| av播播在线观看一区| 成人性生交大片免费视频hd| 亚洲精品日韩在线中文字幕| 人妻少妇偷人精品九色| av网站免费在线观看视频 | 午夜免费观看性视频| 亚洲av电影在线观看一区二区三区 | 午夜精品一区二区三区免费看| 欧美精品国产亚洲| 免费人成在线观看视频色| 丰满乱子伦码专区| 精品少妇黑人巨大在线播放| 男女视频在线观看网站免费| 全区人妻精品视频| 亚洲最大成人手机在线| 国产精品一区二区性色av| 精品欧美国产一区二区三| 你懂的网址亚洲精品在线观看| 免费观看无遮挡的男女| 看免费成人av毛片| 亚洲aⅴ乱码一区二区在线播放| 日本熟妇午夜| 亚洲最大成人av| 国产激情偷乱视频一区二区| 亚洲av.av天堂| 国产 一区 欧美 日韩| 国产成人免费观看mmmm| 在线观看人妻少妇| 中文精品一卡2卡3卡4更新| 成人综合一区亚洲| 国产黄a三级三级三级人| 青春草视频在线免费观看| 国产av不卡久久| 80岁老熟妇乱子伦牲交| 一边亲一边摸免费视频| 联通29元200g的流量卡| or卡值多少钱| 国产精品99久久久久久久久| 美女xxoo啪啪120秒动态图| av在线亚洲专区| 超碰av人人做人人爽久久| 高清视频免费观看一区二区 | 亚洲丝袜综合中文字幕| 精品一区在线观看国产| 亚洲性久久影院| 久久6这里有精品| 女人久久www免费人成看片| 亚洲精品影视一区二区三区av| 白带黄色成豆腐渣| 午夜老司机福利剧场| 在线天堂最新版资源| 国内精品宾馆在线| 身体一侧抽搐| 久久人人爽人人片av| 国产精品无大码| 午夜福利成人在线免费观看| 色综合亚洲欧美另类图片| 国产综合懂色| 国产精品人妻久久久影院| 99久久精品一区二区三区| 久久久久久久午夜电影| 中文资源天堂在线|