• <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 ?

    Acellular allogeneic nerve grafting combined with bone marrow mesenchymal stem cell transplantation for the repair of long-segment sciatic nerve defects: biomechanics and validation of mathematical models

    2016-12-01 09:23:39YajunLiBaolinZhaoHaozeLvZhigangQinMinLuoSchoolofMathematicsJilinUniversityChangchunJilinProvinceChinaDepartmentofOrthopedicsChinaJapanUnionHospitalofJilinUniversityChangchunJilinProvinceChinaDepartmentofClinicalMedi
    關(guān)鍵詞:濾餅濾液巖心

    Ya-jun Li, Bao-lin Zhao, Hao-ze Lv, Zhi-gang Qin, Min Luo School of Mathematics, Jilin University, Changchun, Jilin Province, China Department of Orthopedics, China-Japan Union Hospital of Jilin University, Changchun, Jilin Province, China Department of Clinical Medicine, School of Clinical Medicine, Jilin University, Changchun, Jilin Province, China4 Department of Neurosurgery, China-Japan Union Hospital of Jilin University, Changchun, Jilin Province, China Department of Pain, China-Japan Union Hospital of Jilin University, Changchun, Jilin Province, China

    Acellular allogeneic nerve grafting combined with bone marrow mesenchymal stem cell transplantation for the repair of long-segment sciatic nerve defects: biomechanics and validation of mathematical models

    Ya-jun Li1, Bao-lin Zhao2, Hao-ze Lv3, Zhi-gang Qin4,*, Min Luo5
    1 School of Mathematics, Jilin University, Changchun, Jilin Province, China
    2 Department of Orthopedics, China-Japan Union Hospital of Jilin University, Changchun, Jilin Province, China
    3 Department of Clinical Medicine, School of Clinical Medicine, Jilin University, Changchun, Jilin Province, China
    4 Department of Neurosurgery, China-Japan Union Hospital of Jilin University, Changchun, Jilin Province, China
    5 Department of Pain, China-Japan Union Hospital of Jilin University, Changchun, Jilin Province, China

    How to cite this article: Li YJ, Zhao BL, Lv HZ, Qin ZG, Luo M (2016) Acellular allogeneic nerve grafting combined with bone marrow mesenchymal stem cell transplantation for the repair of long-segment sciatic nerve defects∶ biomechanics and validation of mathematical models. Neural Regen Res 11(8)∶1322-1326.

    Funding: This study was supported by the Science and Technology Development Plan Project Fund of Jilin Province in China, No. 20110492.

    Zhi-gang Qin, M.D.,

    lyj@jlu.edu.cn.

    orcid:

    0000-0002-8080-5241

    (Ya-jun Li)

    Accepted: 2016-05-27

    Graphical Abstract

    We hypothesized that a chemically extracted acellular allogeneic nerve graft used in combination with bone marrow mesenchymal stem cell transplantation would be an effective treatment for long-segment sciatic nerve defects. To test this, we established rabbit models of 30 mm sciatic nerve defects, and treated them using either an autograft or a chemically decellularized allogeneic nerve graft with or without simultaneous transplantation of bone marrow mesenchymal stem cells. We compared the tensile properties, electrophysiological function and morphology of the damaged nerve in each group. Sciatic nerves repaired by the allogeneic nerve graft combined with stem cell transplantation showed better recovery than those repaired by the acellular allogeneic nerve graft alone, and produced similar results to those observed with the autograft. These findings confirm that a chemically extracted acellular allogeneic nerve graft combined with transplantation of bone marrow mesenchymal stem cells is an effective method of repairing long-segment sciatic nerve defects.

    nerve regeneration; chemically extracted acellular allogeneic nerve graft; autograft; bone marrow mesenchymal stem cells; sciatic nerve defects; biomechanics; electrophysiology; morphology; neural regeneration

    Introduction

    Autologous nerve transplantation is considered the gold standard for repairing peripheral nerve damage; however, because of the inevitable damage to other (relatively minor) nerve branches that occurs using this method, allografts are the preferred method used in the clinic (Yu et al., 2014). Although allografts are readily obtainable, immunological rejection is common (Jensen et al., 2005; Zhang et al., 2014a, b). Several techniques have been used to minimize rejection of grafts, including the development of acellular nerves (Zhang et al., 2015a). Zhang et al. (2012) prepared acellular allogeneic nerves by chemical processing, effectively eliminating the immunogenic components of the allograft while retaining Schwann cells, the basement membrane and the integrityof the acellular nerve structure. Zhou et al. (2015) repaired sciatic nerve defects using bone marrow mesenchymal stem cells (BMSCs) combined with grafting of tissue-engineered artificial nerves. The recovery rates of sciatic functional index, nerve conduction and wet weight of triceps muscle were markedly improved, indicating that this technique effectively promotes nerve regeneration and functional recovery. Zhao et al. (2011) demonstrated that chemically extracted acellular nerve allografts (CEANAs) with BMSCs embedded in fibrin glue successfully repaired transected sciatic nerves.

    However, many studies have focused only on the mechanical properties of CEANAs, without evaluating the biomechanical properties after transplantation. We hypothesized that a CEANA would restore the mechanical properties of injured sciatic nerves and thus provide a biomechanical basis for the repair of a sciatic nerve defect.

    Materials and Methods

    Ethical approval

    The experiment was approved by the Animal Ethics Committee of the China-Japan Union Hospital of Jilin University, China. Precautions were taken to minimize suffering and the number of animals used in each experiment.

    Animals

    Seventy-one clean, healthy, female Japanese rabbits, aged 5 months and weighing 2.8-3.1 kg, were provided by the Changchun High-tech Medical Animal Experimental Center, China (licence No. SCXK (Ji) 2003-0004). Rabbits were housed in individual cages at 22-24°C and relative humidity of 56-69%, with air circulation and natural lighting. Rabbits were allowed free access to food (nutritionally complete pellet feeds) and water in their home cages.

    Harvesting sciatic nerves for allogeneic grafts

    Of the 71 rabbits, 20 were selected at random, anesthetized with 10% chloral hydrate (3 mL/kg intraperitoneally), and secured on a surgical table in the supine position. A median incision was made along the posterior part of the left femur. The skin and subcutaneous tissue were cut to dissociate the semimembranosus and semitendinosus muscles and expose the sciatic nerves. A 30-mm segment of sciatic nerve was collected from each rat, bilaterally, at the level of the lower edge of the piriformis (40 segments in total). Specimen dimensions were measured with a reading microscope (CGH-3; Changchun Third Optical Instrument Factory, Changchun, Jilin Province, China). All samples were 30 mm in length and 1.48-1.52 mm in diameter.

    Chemical decellularization of allogeneic nerve

    In accordance with a previous study (Dachtler et al., 2011), sciatic nerve samples were rinsed in distilled water for 1.5 hours, gently rocked in 0.3% Triton X-100 solution for 1.5 hours, and washed three times with distilled water. Sodium deoxycholate solution (0.4%; Shanghai Mingbo Biological Technology Co., Ltd., Shanghai, China) was then added and gentle rocking continued for 1.5 hours. After three further washes with distilled water, the samples were placed in sterile phosphate-buffered saline (PBS; pH 7.4), irradiated with60Co (25 kGy) for 12 hours, and stored at 4°C.

    BMSC culture

    Third and fourth passage mouse BMSCs (Shanghai Yiyan Biological Technology Co., Ltd., Shanghai, China) were placed in basic medium (Shanghai Yiyan Biological Technology Co., Ltd.) containing 20% fetal bovine serum and 50 mL double monoclonal antibodies (penicillin and streptomycin, each 1 × 104U/mL), and incubated at 5% CO2, 37°C, and saturated humidity.

    Preparation of animal models of sciatic nerve defect

    The remaining 51 rabbits were equally and randomly allocated to three groups: autograft, CEANA, and CEANA + BMSCs (n = 17 per group). Rabbits in each group were anesthetized with 6% chloral hydrate (6 mL/kg intraperitoneally) and secured on a surgical table. A median incision was made along the posterior part of the left femur. The skin and subcutaneous tissue were cut to dissociate the semimembranosus and semitendinosus muscles and expose the sciatic nerves bilaterally. A 30-mm segment was excised from each side, 3 mm from the lower edge of the piriformis. The right sciatic nerve from all animals comprised the normal control group.

    Nerve graft repair

    In the autograft group, under an operating microscope (Shanghai Anxin Optical Instrument Co., Ltd., Shanghai, China), the autologous sciatic nerve was turned over and inserted back into the defect, and the epineurium sutured using four 9-0 noninvasive sutures (Qingdao Nesco Medical Co., Ltd., Qingdao, Shandong Province, China) at each end of the graft. Muscle and skin were then sutured.

    The anastomotic stoma was washed with gentamicin and the incision was closed in each group. No external fixation was given after surgery. When the rabbits regained consciousness, they were placed back in individual cages with food and water freely available, and injected intraperitoneally with penicillin (1 × 104U/kg) twice a day for 7 consecutive days. The incision was disinfected with 75% ethanol once a day during this period.

    Electrophysiology

    從表6可以看出,HRD體系無論靜失水和動失水都不大,說明進(jìn)入儲層的濾液少。在巖心靜失水和動失水過程中,巖心可形成薄而密的濾餅,表明具有良好的造壁性。

    Twenty-four weeks after surgery, electrical activity was evaluated by electromyography with a NIM-Neuro 2.0 Nerve Monitor (Medtronic, Minneapolis, MN, USA). Seventeen rabbits from each group were anesthetized with 10% chloral hydrate (400 mg/kg intraperitoneally). In the prone position, the sciatic nerve trunk was exposed bilaterally. The soleus muscle belly was punctured with concentric needleelectrodes, used as recording electrodes. An alligator clip fastened to the skin at the edge of the wound served as the ground wire. Parallel stimulating electrodes were placed at the level of sciatic nodules proximal to the anastomotic stoma and at the sciatic nerve branch distal to the anastomotic stoma to evoke two motor potentials at 50 mA. Electromyography was used to display the amplitude and latency of the action potential. The distance between two stimulating electrodes was measured with a vernier caliper (Shanghai Measuring & Cutting Tool Works Co., Ltd., Shanghai, China). Motor nerve conduction velocity (MNCV) was calculated.

    Table 2 Effects of CEANA combined with BMSC transplantation on tensile properties of sciatic nerves after long-segment damage

    Figure 1 Effects of CEANA combined with BMSC transplantation on sciatic nerve morphology after long-segment damage (hematoxylin-eosin staining, × 400).

    Table 1 Effects of CEANA combined with BMSC transplantation on electrophysiological function of sciatic nerve after long-segment damage

    Sample collection

    After electromyography, a 20 mm length of sciatic nerve was collected from each group (using the anastomotic stoma as the midpoint), and placed in a glass trough containing physiological saline. Fifteen samples from each group were used for tensile testing, and two samples from each group for microstructural observation.

    Figure 2 Tensile stress-strain curves of sciatic nerves in each group. CEANA: Chemically extracted acellular nerve allografts; BMSCs: bone marrow mesenchymal stem cells.

    Hematoxylin-eosin staining

    The sciatic nerve of two rabbits from each group was frozen and cut into 0.8-mm-thick sections, which were fixed in paraformaldehyde for 5 minutes, stained with hematoxylin for 2-5 minutes, treated with HCl-ethanol and then with NaOH, and counterstained with eosin for 20 seconds to 3 minutes; the sections were washed under running tap water after each step. Sections were then dehydrated through a graded alcohol series, permeabilized with xylene, mounted with neutral resin, and observed under a light microscope (Olympus, Tokyo, Japan).

    Tensile testing

    In accordance with previous studies (Jin et al., 2015; Wang et al., 2015; Zhang et al., 2015b), after presetting (loading and unloading were repeated 20 times in each sample),tensile testing was performed in 15 rabbits from each group with an electronic universal testing machine (MODEL55100; Changchun Testing Machine Institute, Changchun, Jilin Province, China). Samples in each group were tested at 36.5 ± 1°C and 2 mm/min. To keep the samples wet, they were sprayed with physiological saline. Tensile stressstrain curves and tensile test data were output by the machine.

    Statistical analysis

    Data are expressed as the mean ± SD and were analyzed using SPSS 16.0 software (SPSS, Chicago, IL, USA). The differences in intergroup data were compared with one-way analysis of variance followed by Scheffe’s method. A value of P < 0.05 was considered statistically significant.

    Results

    Action potential amplitude and MNCV were not significantly different between the autograft and CEANA + BMSCs groups (P > 0.05), but were higher in both of these groups than in the CEANA group (P < 0.05; Table 1).

    CEANA combined with BMSC transplantation improved morphological recovery from long-segment sciatic nerve defect

    Hematoxylin-eosin staining revealed distinct axons surrounded by myelin sheath, and regularly distributed nerve fibers, in the normal control group (Figure 1A). The CEANA + BMSCs group showed good myelinization and regular nerve fibers (Figure 1B). In the CEANA group, most sciatic nerve fibers were regularly arranged, although a few were not (Figure 1C). In the autograft group, nerve fibers were also regularly distributed, and a large amount of myelin sheath was seen in the distal stump of the injured nerve (Figure 1D).

    CEANA combined with BMSC transplantation improved the tensile properties of sciatic nerve after injury

    Tensile testing demonstrated that the elastic limit load, stress and strain, and maximum load, stress and strain, in the CEANA + BMSCs group were higher than in the CEANA group (P < 0.05), but not significantly different from the autograft group (P > 0.05; Table 2).

    Stress-strain curves and functions for stress-strain relationship

    Stress-strain curves for sciatic nerve samples in each group were drawn (Figure 2), and curve fitting was conducted on tensile testing data. Stress-strain curves showed exponential changes when sciatic nerve strain increased from 0 to 7.06%, 6.32%, 5.11% and 6.41% in the normal control, CEANA, CEANA + BMSCs, and autograft groups, respectively. A linear stress-strain relationship was observed when sciatic nerve strain increased from 7.07% to 14.86%, from 6.33% to 13.01.%, from 5.12% to 11.31%, and from 6.42% to 13.32% in the normal control, CEANA, CEANA + BMSCs and autograft groups, respectively. When sciatic nerve strain increased from 14.87% to 23.23%, from 13.02% to 21.9%, from 11.32% to 19.20%, and from 13.33% to 21.86% in the normal control, CEANA, CEANA + BMSCs, and autograft groups, respectively, samples showed marked deformation, near loss of bearing capacity, and damage.

    Functions for sciatic nerve stress-strain relationships (σ(ε)) were constructed by linear regression analysis in each group, as follows: normal control group, σ(ε) = 0.0988e5+ 0.7307e4+ 2.6155e3- 0.1431e2; CEANA + BMSCs group, σ(ε) = 0.0745e5+ 0.5970e4+ 2.4617e3- 0.0907e2; autograft group, σ(ε) = 0.1739e5+ 1.344e4+ 0.7573e3+ 0.6122e2; CEANA group, σ(ε) = 0.09324e5+ 0.8136e4+ 0.5798e3+ 0.2637e2.

    Discussion

    The biomechanical properties of peripheral nerves are maintained by surrounding connective tissue, of which collagen fiber is the main component. Collagen is tough, with high tensile strength, and can withstand a certain amount of mechanical stimulation. Its quantity and distribution determine the biomechanical properties of peripheral nerves (Eather et al., 1986). Chemically extracted acellular allogeneic nerve is a new tissue-engineered material with low immunogenicity and a three-dimensional structure (Sondell et al., 1998; Hudson et al., 2004). CEANAs make use of this material to guide Schwann cell migration and promote axonal regeneration, and are a promising substitute for autologous nerve transplantation. Borschel et al. (2003) confirmed that nerve decellularization processes may remove one or more collagen components, leading to changes in the mechanical properties of the nerve. He et al. (2009) found that tissue engineered nerves constructed with BMSCs had better reparative effects in 10 mm sciatic nerve defects than did CEANAs. BMSCs can differentiate into neural cells, replace apoptotic nerve cells, secrete neurotrophic factors, and promote axonal regeneration. BMSCs can also regulate Schwann cells and promote peripheral nerve regeneration, and are ideal seed cells (Lin et al., 2008; Wang et al., 2009; Zheng et al., 2010).

    Greater CMAP and MNCV are associated with better recovery of tissue morphology and stronger nerves after repair. In addition, the injury mechanism and functional recovery of the sciatic nerve were strongly associated with its mechanical properties. The present findings indicate that CEANA combined with BMSC transplantation markedly improved sciatic nerve recovery compared with CEANA alone, to a degree similar to that after an autograft. CEANA combined with BMSC transplantation is a promising treatment for the repair of peripheral nerve damage in the clinic. Further research to improve the method and its introduction in the clinic will identify additional applications for this technique.

    The present results demonstrate that CEANA used in combination with BMSC transplantation for the repair of sciatic nerve defects restores damaged collagen and improves the biomechanical properties of the sciatic nerve. Furthermore, CEANA combined with BMSC transplantation enhanced the electrophysiological properties of the sciatic nerve after injury. We also calculated the stress-strain function in thedamaged nerves using regression analysis. Experimental data were evaluated using mathematical and statistical models, to better understand the mechanical properties of the repaired nerve.

    Because of individual differences and the limited number of experimental animals, there is a large amount of dispersion among the experimental data. However, the present data provide a valuable reference for further investigation into the treatment of sciatic nerve injury.

    Author contributions: YJL and ZGQ conceived and designed the study. HZL provided the data. BLZ and ZGQ ensured the integrity of the data and analyzed data. YJL wrote the paper and served as a principle investigator. ZGQ was in charge of manuscript authorization. BLZ undertook the statistical analysis. ML obtained funding. ML and BLZ provided technical or data support. All authors performed the experiments, and approved the final version of the paper.

    Conflicts of interest: None declared.

    Plagiarism check: This paper was screened twice using Cross-Check to verify originality before publication.

    Peer review: This paper was double-blinded and stringently reviewed by international expert reviewers.

    References

    Borschel GH, Kia KF, Kuzon WM Jr, Dennis RG (2003) Mechanical properties of acellular peripheral nerve. J Surg Res 114:133-139.

    Dachtler J, Hardingham NR, Glazewski S, Wright NF, Blain EJ, Fox K (2011) Experience-dependent plasticity acts via GluR1 and a novel αNOS1 dependent synaptic mechanism in adult cortex. J Neurosci 31:11220-11230.

    Eather TF, Pollock M, Myers DB (1986) Proximal and distal changes in collagen content of peripheral nerve that follow transection and crush lesions. Exp Neurol 92:299-310.

    He HY, Deng YH, Tong XJ, Cheng JM, Du ZK (2009) Repair of sciatic nerve defects with tissue engineered nerves constructed with marrow stromal cells. Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu 13:5662-5566.

    Hudson TW, Liu SY, Schmidt CE (2004) Engineering an improved acellular nerve graft via optimized chemical processing. Tissue Eng 10:1346-1358.

    Jensen JN, Brenner MJ, Tung TH, Hunter DA, Mackinnon SE (2005) Effect of FK506 on peripheral nerve regeneration through long grafts in inbred swine. Ann Plast Surg 54:420-427.

    Jin H, Yang Q, Ji F, Zhang YJ, Zhao Y, Luo M (2015) Human amniotic epithelial cell transplantation for the repair of injured brachial plexus nerve: evaluation of nerve viscoelastic properties. Neural Regen Res 10:260-265.

    Lin W, Chen X, Wang X, Liu J, Gu X (2008) Adult rat bone marrow stromal cells differentiate into Schwann cell-like cells in vitro. In Vitro Cell Dev Biol Anim 44:31-40.

    Ma XL, Yang ZB, Li XL, Ma JX, Zhang Y, Guo HG, Sun XL (2010) A study on biomechanical properties of chemically extracted acellular peripheral nerve. Zhongguo Xiufu Chongjian Waike Zazhi 24:1293-1297.

    Sondell M, Lundborg G, Kanje M (1998) Regeneration of the rat sciatic nerve into allografts made acellular through chemical extraction. Brain Res 795:44-54.

    Wang J, Ding F, Gu Y, Liu J, Gu X (2009) Bone marrow mesenchymal stem cells promote cell proliferation and neurotrophic function of Schwann cells in vitro and in vivo. Brain Res 1262:7-15.

    Wang Y, Li ZW, Luo M, Li YJ, Zhang KQ (2015) Biological conduits combining bone marrow mesenchymal stem cells and extracellular matrix to treat long-segment sciatic nerve defects. Neural Regen Res 10:965-971.

    Yang Z, Ma XL, Li XL, Ma JX, Zhang Y, Guo HG, Sun XL (2011) Effects of different acellular methods on biomechanical properties of peripheral nerve. Zhongguo Shengwu Yixue Gongcheng Xuebao 30:155-159.

    Yu GM, Wang W, Zhang L, Zhang DL (2014) Repairing sciatic nerve in rats by acellular allogeneic nerve transplantation treated with chondroitinase ABC-PLGA microspheres. Jiefangjun Yixueyuan Xuebao 35:858-862.

    Zhang Y, Zhang H, Katiella K, Huang W (2014a) Chemically extracted acellular allogeneic nerve graft combined with ciliary neurotrophic factor promotes sciatic nerve repair. Neural Regen Res 9:1358-1364. Zhang Y, Zhang H, Zhang G, Ka K, Huang W (2014b) Combining acellular nerve allografts with brain-derived neurotrophic factor transfected bone marrow mesenchymal stem cells restores sciatic nerve injury better than either intervention alone. Neural Regen Res 9:1814-1819.

    Zhang YR, Yao JN, Zhou MW, Li YB, Wang YS (2012) Human placenta amniotic membrane wrap chemically extracted acellular nerve allograft repair of canine nerves. Zhengzhou Daxue Xuebao: Yixue Ban 47:509-511.

    Zhang YR, Ka K, Zhang GC, Zhang H, Shang Y, Zhao GQ, Huang WH (2015a) Repair of peripheral nerve defects with chemically extracted acellular nerve allografts loaded with neurotrophic factors-transfected bone marrow mesenchymal stem cells. Neural Regen Res 10:1498-1506.

    Zhang ZJ, Li YJ, Liu XG, Huang FX, Liu TJ, Jiang DM, Lv XM, Luo M (2015b) Human umbilical cord blood stem cells and brain-derived neurotrophic factor for optic nerve injury: a biomechanical evaluation. Neural Regen Res 10:1134-1138.

    Zhao Z, Wang Y, Peng J, Zhao B, Zhao Q, Liu Y, Ren ZW, Zhan SF, Zhang L, Xu WJ, Lu SB (2011) Effect of chemical extracted acellular nerve allograft supplementing with bone marrow mesenchymal stem cells embedded in fibrin glue on functional recovery of transected sciatic nerves. Zhongguo Xiufu Chongjian Waike Zazhi 25:488-493.

    Zheng W, Honmou O, Miyata K, Harada K, Suzuki J, Liu H, Houkin K, Hamada H, Kocsis JD (2010) Therapeutic benefits of human mesenchymal stem cells derived from bone marrow after global cerebral ischemia. Brain Res 1310:8-16.

    Zhou LN, Cui XJ, Zu KX, Wang XH, Cai XY, Guo JH, Na QQ (2015) Repairment of 1 cm sciatic nerve defect by bone marrow mesenchymal stem cell of adult rat combined with tissue-engineered artificial nerve. Qiguan Yizhi 6:157-160.

    Copyedited by Slone-Murphy J, Haase R, Yu J, Qiu Y, Li CH, Song LP, Zhao M

    10.4103/1673-5374.189198

    The CEANA group an acellular graft, which was also turned over and sutured. In the CEANA + BMSCs group, fifth passage mouse BMSCs (1 mL; approximately 5 × 105) were infused into the CEANA conduit, and the sciatic nerve tissue was turned over and sutured.

    *Correspondence to:

    猜你喜歡
    濾餅濾液巖心
    長填齡滲濾液MBR+NF組合工藝各處理單元的DOM化學(xué)多樣性
    Osteotomized folded scapular tip free flap for complex midfacial reconstruction
    等壓濾餅過濾過程中五個基本理論問題之探討
    ——非均布濾餅的局部比阻與平均比阻的測定與計算方法
    某滲濾液收集池底部防滲層鼓包的分析與治理
    雙級推料離心機應(yīng)用在薄層濾餅上的結(jié)構(gòu)優(yōu)化
    流體機械(2020年5期)2020-06-24 05:39:08
    進(jìn)水pH對MBR處理垃圾滲濾液效果的影響
    南蘇丹Palouge油田濾餅清除技術(shù)
    一種頁巖巖心資料的保存方法
    化工管理(2017年23期)2017-09-11 14:14:22
    濃度和粒度對細(xì)粒煤濾餅結(jié)構(gòu)影響的研究
    DTRO工藝處理垃圾滲濾液的研究
    五月玫瑰六月丁香| 男女边吃奶边做爰视频| 最新中文字幕久久久久| 我要看日韩黄色一级片| 秋霞在线观看毛片| 国产探花在线观看一区二区| 国产精品久久久久久久电影| 免费av毛片视频| 69人妻影院| 久久久久性生活片| 在线国产一区二区在线| 此物有八面人人有两片| 久久久久久大精品| 国产日本99.免费观看| 美女被艹到高潮喷水动态| 亚洲第一区二区三区不卡| 欧美在线一区亚洲| 国产精品久久久久久精品电影小说 | 国产精品女同一区二区软件| 欧美丝袜亚洲另类| 免费av观看视频| 国产一级毛片七仙女欲春2| 久久久成人免费电影| 69av精品久久久久久| 国产精品1区2区在线观看.| 国产亚洲av片在线观看秒播厂 | 少妇被粗大猛烈的视频| 长腿黑丝高跟| 久久久国产成人免费| 天堂影院成人在线观看| 插逼视频在线观看| av免费在线看不卡| 一级毛片我不卡| 久久草成人影院| 亚洲精华国产精华液的使用体验 | 麻豆精品久久久久久蜜桃| 亚洲人成网站在线播| 亚洲无线观看免费| av在线观看视频网站免费| 亚洲av免费在线观看| 卡戴珊不雅视频在线播放| 国产精品伦人一区二区| 日韩,欧美,国产一区二区三区 | 青春草亚洲视频在线观看| 国产精品一区www在线观看| 日韩欧美国产在线观看| 夜夜看夜夜爽夜夜摸| 久久精品夜夜夜夜夜久久蜜豆| 我要搜黄色片| 欧美极品一区二区三区四区| 亚洲经典国产精华液单| 亚洲综合色惰| 禁无遮挡网站| 亚洲三级黄色毛片| 国产爱豆传媒在线观看| 国产精品爽爽va在线观看网站| 人妻少妇偷人精品九色| 午夜视频国产福利| 此物有八面人人有两片| 在线观看美女被高潮喷水网站| 狂野欧美白嫩少妇大欣赏| 99热精品在线国产| 中文字幕av成人在线电影| 麻豆国产97在线/欧美| 美女cb高潮喷水在线观看| 在线播放国产精品三级| 亚洲国产高清在线一区二区三| 99久久精品热视频| 国内精品宾馆在线| 欧美潮喷喷水| av天堂在线播放| 色综合站精品国产| 国产视频内射| 九草在线视频观看| 美女国产视频在线观看| 一本一本综合久久| 欧美变态另类bdsm刘玥| 亚洲乱码一区二区免费版| 久久久精品欧美日韩精品| 午夜激情福利司机影院| 久久久久久久久久成人| 日韩欧美精品v在线| 黄色配什么色好看| 久久99热这里只有精品18| 免费搜索国产男女视频| 亚洲天堂国产精品一区在线| 我的老师免费观看完整版| 波多野结衣高清作品| 久久午夜亚洲精品久久| 看黄色毛片网站| 22中文网久久字幕| 我要搜黄色片| 国内久久婷婷六月综合欲色啪| 看十八女毛片水多多多| 国产私拍福利视频在线观看| 国产片特级美女逼逼视频| 亚洲在线观看片| 亚洲欧美精品自产自拍| 久久精品综合一区二区三区| 亚洲一区高清亚洲精品| 久久久久国产网址| 色吧在线观看| 男人和女人高潮做爰伦理| 大又大粗又爽又黄少妇毛片口| 夜夜看夜夜爽夜夜摸| 黑人高潮一二区| 亚洲最大成人手机在线| 午夜免费男女啪啪视频观看| 亚洲精品成人久久久久久| 国产精品三级大全| 欧美性猛交╳xxx乱大交人| 久久精品夜夜夜夜夜久久蜜豆| 99热这里只有是精品50| 亚洲精品乱码久久久v下载方式| 直男gayav资源| 精品日产1卡2卡| 免费av观看视频| 欧美精品一区二区大全| 男人舔女人下体高潮全视频| 熟女电影av网| 又粗又爽又猛毛片免费看| av在线蜜桃| 久久久精品大字幕| 久久人人爽人人片av| 一本久久中文字幕| 亚洲七黄色美女视频| 国产一区亚洲一区在线观看| 大型黄色视频在线免费观看| 尾随美女入室| 成人亚洲精品av一区二区| 国产真实乱freesex| 午夜精品在线福利| 18禁在线无遮挡免费观看视频| 狠狠狠狠99中文字幕| 午夜亚洲福利在线播放| 三级毛片av免费| 精品久久久久久久久久免费视频| 人人妻人人澡人人爽人人夜夜 | 身体一侧抽搐| 国产黄片美女视频| 永久网站在线| 两性午夜刺激爽爽歪歪视频在线观看| 国产大屁股一区二区在线视频| 色5月婷婷丁香| 欧美日韩国产亚洲二区| 又爽又黄a免费视频| 国产中年淑女户外野战色| 久久精品国产99精品国产亚洲性色| 欧美色视频一区免费| 亚洲国产精品成人久久小说 | 亚洲成人av在线免费| 日本-黄色视频高清免费观看| 成人特级黄色片久久久久久久| 少妇熟女aⅴ在线视频| 啦啦啦韩国在线观看视频| 啦啦啦韩国在线观看视频| 久久这里有精品视频免费| 国产亚洲欧美98| 一级毛片我不卡| 99久久精品国产国产毛片| 国产淫片久久久久久久久| 亚洲国产精品成人综合色| 69人妻影院| 最近2019中文字幕mv第一页| 亚洲欧美中文字幕日韩二区| 青春草亚洲视频在线观看| 99久久中文字幕三级久久日本| 日韩制服骚丝袜av| 噜噜噜噜噜久久久久久91| 我要看日韩黄色一级片| 成人午夜高清在线视频| 99riav亚洲国产免费| 日韩av在线大香蕉| 日日干狠狠操夜夜爽| 欧美性感艳星| 99久久无色码亚洲精品果冻| 男人和女人高潮做爰伦理| a级毛色黄片| 精品人妻一区二区三区麻豆| 啦啦啦观看免费观看视频高清| 欧美日本视频| 在线观看66精品国产| av在线观看视频网站免费| 色吧在线观看| 亚洲av一区综合| 给我免费播放毛片高清在线观看| 晚上一个人看的免费电影| 99热精品在线国产| av在线亚洲专区| 草草在线视频免费看| 精品欧美国产一区二区三| 日韩成人伦理影院| 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 | 久久久久国产网址| 亚洲国产欧美人成| 天堂影院成人在线观看| 欧美日韩在线观看h| 亚洲五月天丁香| 男女那种视频在线观看| 麻豆成人av视频| h日本视频在线播放| 国产精品一区二区性色av| 插逼视频在线观看| 婷婷精品国产亚洲av| 午夜福利在线在线| 久久99热这里只有精品18| 综合色av麻豆| 久久国内精品自在自线图片| 干丝袜人妻中文字幕| 久久人妻av系列| 亚洲,欧美,日韩| 国产高潮美女av| 亚洲av.av天堂| 亚洲av一区综合| 国产精华一区二区三区| 久久久久性生活片| 久久精品国产亚洲网站| 亚洲久久久久久中文字幕| 久久久a久久爽久久v久久| 日韩一本色道免费dvd| 九九热线精品视视频播放| av卡一久久| 最近2019中文字幕mv第一页| 色5月婷婷丁香| 亚洲精品日韩av片在线观看| 男女边吃奶边做爰视频| 嫩草影院入口| 中文字幕人妻熟人妻熟丝袜美| 两性午夜刺激爽爽歪歪视频在线观看| 大型黄色视频在线免费观看| a级毛片免费高清观看在线播放| 成人性生交大片免费视频hd| 中文字幕久久专区| 我的老师免费观看完整版| 亚洲内射少妇av| 日韩 亚洲 欧美在线| 久久久久久久久中文| 久久久午夜欧美精品| 嫩草影院精品99| 久久精品国产99精品国产亚洲性色| 天天一区二区日本电影三级| 久久99热这里只有精品18| 毛片女人毛片| 国产在线男女| 欧美日本亚洲视频在线播放| 久久精品综合一区二区三区| 1024手机看黄色片| 国产高清三级在线| av在线蜜桃| 丰满乱子伦码专区| 中文字幕精品亚洲无线码一区| 亚洲欧美成人精品一区二区| 自拍偷自拍亚洲精品老妇| 69人妻影院| 亚洲不卡免费看| 黄色一级大片看看| 精品国产三级普通话版| 日韩欧美精品免费久久| 99热这里只有是精品50| 国产精品综合久久久久久久免费| 亚洲最大成人av| 免费看日本二区| 国产精品1区2区在线观看.| 九九久久精品国产亚洲av麻豆| 亚洲国产精品成人综合色| 亚洲精品乱码久久久v下载方式| 美女脱内裤让男人舔精品视频 | 亚洲精品国产av成人精品| 午夜精品一区二区三区免费看| 久久精品人妻少妇| 噜噜噜噜噜久久久久久91| 久久草成人影院| 黄色日韩在线| 国产精品人妻久久久影院| 乱系列少妇在线播放| 99热精品在线国产| 免费看a级黄色片| av卡一久久| 天堂影院成人在线观看| 国内揄拍国产精品人妻在线| 免费看日本二区| 亚洲精品乱码久久久久久按摩| 18禁裸乳无遮挡免费网站照片| 国产精品99久久久久久久久| 欧美性猛交╳xxx乱大交人| 国产探花在线观看一区二区| av又黄又爽大尺度在线免费看 | 国产探花在线观看一区二区| 亚洲自偷自拍三级| 国产精品永久免费网站| 男女下面进入的视频免费午夜| 久久久久性生活片| 2022亚洲国产成人精品| 麻豆国产av国片精品| 18+在线观看网站| 欧美一区二区精品小视频在线| 日韩中字成人| 国产成年人精品一区二区| 偷拍熟女少妇极品色| 天天躁日日操中文字幕| 国产精华一区二区三区| 能在线免费观看的黄片| 午夜精品国产一区二区电影 | 最近的中文字幕免费完整| 国产综合懂色| 亚洲在久久综合| 91av网一区二区| 亚洲国产色片| 国产女主播在线喷水免费视频网站 | 亚洲自拍偷在线| 中文字幕精品亚洲无线码一区| 精品一区二区三区人妻视频| 亚洲av免费高清在线观看| 人体艺术视频欧美日本| 美女内射精品一级片tv| 亚洲av二区三区四区| 最后的刺客免费高清国语| 两性午夜刺激爽爽歪歪视频在线观看| 日韩成人av中文字幕在线观看| 成人午夜精彩视频在线观看| 成年女人永久免费观看视频| 成人欧美大片| 久久久久久久午夜电影| 可以在线观看毛片的网站| 亚洲18禁久久av| 乱码一卡2卡4卡精品| 亚洲精品日韩av片在线观看| 久久久久九九精品影院| 中文资源天堂在线| 亚洲av男天堂| 国产精品久久久久久精品电影小说 | 在线观看66精品国产| 日日摸夜夜添夜夜爱| 天天一区二区日本电影三级| 成人午夜高清在线视频| 久久精品国产亚洲av涩爱 | .国产精品久久| 女人十人毛片免费观看3o分钟| 亚洲性久久影院| 国产精品爽爽va在线观看网站| 免费一级毛片在线播放高清视频| 国产精品三级大全| 天堂av国产一区二区熟女人妻| avwww免费| 老司机影院成人| 国产精品野战在线观看| 国产精品久久久久久亚洲av鲁大| av在线观看视频网站免费| 亚洲无线在线观看| 91久久精品国产一区二区三区| 亚洲18禁久久av| 国产老妇伦熟女老妇高清| 国产精品久久电影中文字幕| 亚洲精华国产精华液的使用体验 | 亚洲国产精品成人久久小说 | 久久精品影院6| 精品日产1卡2卡| 五月伊人婷婷丁香| 欧美高清性xxxxhd video| 男人舔女人下体高潮全视频| 亚洲精品乱码久久久v下载方式| 在线观看一区二区三区| 色尼玛亚洲综合影院| 日韩精品有码人妻一区| av黄色大香蕉| 亚洲18禁久久av| 精品久久久噜噜| 久久久久久久久大av| 亚洲精品456在线播放app| 边亲边吃奶的免费视频| 免费人成视频x8x8入口观看| 少妇的逼水好多| 九九久久精品国产亚洲av麻豆| 国产精品一区www在线观看| 天天一区二区日本电影三级| 国产av不卡久久| 一卡2卡三卡四卡精品乱码亚洲| 国产乱人视频| 一边摸一边抽搐一进一小说| 亚洲成av人片在线播放无| 久久久久久久久久黄片| 黄色欧美视频在线观看| 有码 亚洲区| 大又大粗又爽又黄少妇毛片口| 深夜精品福利| 看片在线看免费视频| 91久久精品国产一区二区三区| kizo精华| 亚洲av一区综合| 亚洲欧美成人精品一区二区| 久久久久久伊人网av| 久久精品久久久久久久性| 在线观看66精品国产| 精品久久国产蜜桃| 99热这里只有精品一区| 日韩精品青青久久久久久| 一区二区三区免费毛片| 在线播放无遮挡| 男女做爰动态图高潮gif福利片| 秋霞在线观看毛片| 神马国产精品三级电影在线观看| 国产单亲对白刺激| 国产高清激情床上av| av黄色大香蕉| 久久精品影院6| 国产精品野战在线观看| 国产精品女同一区二区软件| 国产成人freesex在线| 日韩成人av中文字幕在线观看| 人妻制服诱惑在线中文字幕| 欧美成人一区二区免费高清观看| 国内精品美女久久久久久| 国产人妻一区二区三区在| 亚洲精品亚洲一区二区| 干丝袜人妻中文字幕| eeuss影院久久| 国产精品久久电影中文字幕| 国产蜜桃级精品一区二区三区| 最近最新中文字幕大全电影3| 五月玫瑰六月丁香| 日韩欧美在线乱码| 一区二区三区四区激情视频 | 国产老妇伦熟女老妇高清| 国产乱人视频| 嘟嘟电影网在线观看| 热99在线观看视频| 精品久久久噜噜| 国国产精品蜜臀av免费| av在线亚洲专区| 岛国在线免费视频观看| 国产老妇女一区| 久久久久网色| 日韩一本色道免费dvd| 国产亚洲欧美98| 色噜噜av男人的天堂激情| 精品久久久久久久久久久久久| 久久久久久久久久黄片| 日韩欧美在线乱码| 亚洲aⅴ乱码一区二区在线播放| 一级黄色大片毛片| 国产人妻一区二区三区在| 亚洲真实伦在线观看| 亚洲av不卡在线观看| 日韩制服骚丝袜av| 欧美一级a爱片免费观看看| 亚洲国产欧美人成| 亚洲欧美日韩东京热| 国模一区二区三区四区视频| 我的老师免费观看完整版| 久久精品国产亚洲网站| 久久精品夜夜夜夜夜久久蜜豆| 观看免费一级毛片| a级毛片a级免费在线| 亚洲欧美日韩东京热| 亚洲精品久久国产高清桃花| 日韩欧美 国产精品| 日韩一区二区三区影片| 男女下面进入的视频免费午夜| 最近最新中文字幕大全电影3| 国产精品三级大全| 日韩,欧美,国产一区二区三区 | 国产综合懂色| 亚洲无线在线观看| 欧美不卡视频在线免费观看| 禁无遮挡网站| 可以在线观看毛片的网站| 久久精品国产清高在天天线| 看片在线看免费视频| 天美传媒精品一区二区| 在线天堂最新版资源| 久久九九热精品免费| 久久欧美精品欧美久久欧美| 精品久久久久久成人av| 哪里可以看免费的av片| 久久久久久久久久成人| 最后的刺客免费高清国语| 国产爱豆传媒在线观看| 精品日产1卡2卡| 在线免费观看的www视频| 国产精品野战在线观看| 亚洲欧美日韩卡通动漫| 色哟哟哟哟哟哟| 国产亚洲欧美98| 婷婷六月久久综合丁香| 久久这里有精品视频免费| 嫩草影院新地址| 国产老妇女一区| 亚洲人与动物交配视频| 一区福利在线观看| 99热精品在线国产| а√天堂www在线а√下载| 国产乱人偷精品视频| 天堂√8在线中文| 欧美日韩国产亚洲二区| 亚洲va在线va天堂va国产| 亚洲精品乱码久久久久久按摩| 国产精品伦人一区二区| 美女黄网站色视频| 久久亚洲国产成人精品v| 久久韩国三级中文字幕| 99久久久亚洲精品蜜臀av| 免费看av在线观看网站| 久久久久久久久大av| 欧美变态另类bdsm刘玥| 夜夜爽天天搞| 男女啪啪激烈高潮av片| 欧洲精品卡2卡3卡4卡5卡区| 亚洲18禁久久av| 麻豆成人av视频| 亚洲va在线va天堂va国产| 国产精品久久久久久亚洲av鲁大| 国产久久久一区二区三区| 国产亚洲91精品色在线| 日本熟妇午夜| 久久久久网色| 精品久久久久久久久亚洲| 十八禁国产超污无遮挡网站| 18禁在线播放成人免费| 国产精品久久久久久久电影| 成年免费大片在线观看| 国产精品久久久久久精品电影| 看片在线看免费视频| 色播亚洲综合网| 国产女主播在线喷水免费视频网站 | 国产高潮美女av| 国产亚洲av嫩草精品影院| 日本黄色视频三级网站网址| av卡一久久| 99久久九九国产精品国产免费| 国产精品女同一区二区软件| 麻豆成人av视频| 免费大片18禁| 白带黄色成豆腐渣| 最近的中文字幕免费完整| 男人舔女人下体高潮全视频| 99久久精品一区二区三区| 日韩av在线大香蕉| 中文字幕av成人在线电影| 内地一区二区视频在线| 丰满乱子伦码专区| 国产高清不卡午夜福利| 麻豆精品久久久久久蜜桃| 亚洲av.av天堂| 久久久久免费精品人妻一区二区| 能在线免费看毛片的网站| 蜜臀久久99精品久久宅男| 国产91av在线免费观看| 色哟哟·www| h日本视频在线播放| 最近的中文字幕免费完整| 日韩制服骚丝袜av| 在线播放无遮挡| 一本久久中文字幕| 深夜a级毛片| 成人综合一区亚洲| 日日干狠狠操夜夜爽| 成人毛片a级毛片在线播放| 成人亚洲精品av一区二区| 亚洲欧洲日产国产| av福利片在线观看| 久久99热6这里只有精品| 久久久久久久久久黄片| 亚洲最大成人中文| 亚洲成人久久爱视频| 久久婷婷人人爽人人干人人爱| 国产在线精品亚洲第一网站| 麻豆国产97在线/欧美| 午夜精品一区二区三区免费看| 丰满人妻一区二区三区视频av| 乱码一卡2卡4卡精品| 精品久久久久久久久av| 熟妇人妻久久中文字幕3abv| 午夜老司机福利剧场| 国内精品一区二区在线观看| 在线观看午夜福利视频| av.在线天堂| 1024手机看黄色片| 男女啪啪激烈高潮av片| 少妇熟女欧美另类| 少妇人妻一区二区三区视频| 午夜福利成人在线免费观看| 婷婷六月久久综合丁香| av在线老鸭窝| 亚洲欧洲日产国产| 国产伦在线观看视频一区| 免费搜索国产男女视频| 成人特级黄色片久久久久久久| 国产淫片久久久久久久久| 亚洲精品久久久久久婷婷小说 | 中文字幕制服av| a级毛片免费高清观看在线播放| 九九在线视频观看精品| 国产精品久久久久久久久免| 国产一区二区激情短视频| .国产精品久久| 干丝袜人妻中文字幕| 国产黄a三级三级三级人| 一级黄色大片毛片| 免费看日本二区| 国国产精品蜜臀av免费| 熟女电影av网| 成熟少妇高潮喷水视频| 噜噜噜噜噜久久久久久91| 亚洲欧美精品自产自拍| 内地一区二区视频在线| 深爱激情五月婷婷| 悠悠久久av| 国内久久婷婷六月综合欲色啪| 欧美性猛交╳xxx乱大交人| 1000部很黄的大片| 99久久成人亚洲精品观看| 日本av手机在线免费观看| 一进一出抽搐gif免费好疼| 国产乱人偷精品视频| 一夜夜www|