• <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
    久久精品影院6| 免费久久久久久久精品成人欧美视频| 亚洲精品美女久久av网站| 国产午夜精品久久久久久| 免费女性裸体啪啪无遮挡网站| 久久久久久久久久久久大奶| www.www免费av| 久久精品91无色码中文字幕| 最近最新中文字幕大全免费视频| 69av精品久久久久久| 亚洲国产精品成人综合色| 禁无遮挡网站| tocl精华| 国产亚洲av嫩草精品影院| av有码第一页| 久久九九热精品免费| 老汉色av国产亚洲站长工具| 巨乳人妻的诱惑在线观看| 身体一侧抽搐| 国产亚洲精品综合一区在线观看 | 国产成人精品无人区| 黄频高清免费视频| 精品久久久久久,| 最好的美女福利视频网| 18禁观看日本| 成人国产综合亚洲| 黄片播放在线免费| 日本在线视频免费播放| 日本一区二区免费在线视频| 狠狠狠狠99中文字幕| 人人妻人人爽人人添夜夜欢视频| 国产又色又爽无遮挡免费看| 男女午夜视频在线观看| 国产av在哪里看| 欧美亚洲日本最大视频资源| 亚洲三区欧美一区| 久久草成人影院| avwww免费| 久久久久国产精品人妻aⅴ院| 看免费av毛片| 国产91精品成人一区二区三区| 在线播放国产精品三级| 丝袜人妻中文字幕| 免费看a级黄色片| 18禁裸乳无遮挡免费网站照片 | 在线观看舔阴道视频| 成人手机av| 一级,二级,三级黄色视频| 亚洲国产毛片av蜜桃av| 不卡一级毛片| 最新在线观看一区二区三区| 曰老女人黄片| 国产日韩一区二区三区精品不卡| 99精品久久久久人妻精品| 欧美日本中文国产一区发布| 免费无遮挡裸体视频| 大香蕉久久成人网| 久久人妻福利社区极品人妻图片| 亚洲国产精品sss在线观看| 黄色片一级片一级黄色片| 亚洲无线在线观看| 国产视频一区二区在线看| 日本三级黄在线观看| а√天堂www在线а√下载| 51午夜福利影视在线观看| 嫩草影院精品99| 搡老妇女老女人老熟妇| 欧美日韩乱码在线| 国产精品精品国产色婷婷| 亚洲av熟女| 男人操女人黄网站| 国产成人欧美在线观看| 精品一区二区三区av网在线观看| 咕卡用的链子| 国产精品免费视频内射| 在线观看免费视频日本深夜| 天天一区二区日本电影三级 | 国产亚洲精品一区二区www| 在线观看舔阴道视频| 黄色 视频免费看| 国产xxxxx性猛交| 国产精品免费一区二区三区在线| 久久国产精品男人的天堂亚洲| 12—13女人毛片做爰片一| 亚洲五月色婷婷综合| 丝袜在线中文字幕| 亚洲色图综合在线观看| 国产人伦9x9x在线观看| 国产1区2区3区精品| 一区二区三区激情视频| 亚洲熟妇中文字幕五十中出| 亚洲成av人片免费观看| 精品久久久久久久久久免费视频| 九色亚洲精品在线播放| 伊人久久大香线蕉亚洲五| 欧美老熟妇乱子伦牲交| 一区二区三区精品91| 女人被狂操c到高潮| 国产亚洲精品一区二区www| 91字幕亚洲| 国产成年人精品一区二区| 久久天躁狠狠躁夜夜2o2o| 成年女人毛片免费观看观看9| 看片在线看免费视频| 夜夜看夜夜爽夜夜摸| 一区福利在线观看| 中亚洲国语对白在线视频| 欧美色视频一区免费| 中文字幕最新亚洲高清| 亚洲中文字幕日韩| 日本精品一区二区三区蜜桃| 精品欧美国产一区二区三| av片东京热男人的天堂| 一级毛片高清免费大全| 国产精品久久久av美女十八| 日韩 欧美 亚洲 中文字幕| 女人高潮潮喷娇喘18禁视频| 亚洲国产高清在线一区二区三 | 一区福利在线观看| 91成人精品电影| 99久久久亚洲精品蜜臀av| 看免费av毛片| 啦啦啦观看免费观看视频高清 | 久热爱精品视频在线9| 女性被躁到高潮视频| 不卡av一区二区三区| 很黄的视频免费| 久热爱精品视频在线9| 日韩中文字幕欧美一区二区| 国产乱人伦免费视频| 黄网站色视频无遮挡免费观看| 亚洲精品在线观看二区| 国产精品二区激情视频| cao死你这个sao货| 精品一区二区三区四区五区乱码| 中文字幕最新亚洲高清| 亚洲色图综合在线观看| 国产精品一区二区精品视频观看| 久久久久久免费高清国产稀缺| 国产精品影院久久| 精品人妻1区二区| 久久午夜综合久久蜜桃| 午夜免费鲁丝| 美国免费a级毛片| 日韩中文字幕欧美一区二区| 中文字幕人成人乱码亚洲影| 热re99久久国产66热| 亚洲av电影不卡..在线观看| 色尼玛亚洲综合影院| 高潮久久久久久久久久久不卡| 久久久久久久久中文| 亚洲国产欧美网| 亚洲欧美日韩另类电影网站| 精品人妻1区二区| 亚洲熟妇熟女久久| 欧美一级毛片孕妇| 国产熟女xx| 国产精品乱码一区二三区的特点 | 国产成人免费无遮挡视频| 亚洲va日本ⅴa欧美va伊人久久| 亚洲成人久久性| 国产一区二区三区综合在线观看| 桃色一区二区三区在线观看| 日韩精品中文字幕看吧| 午夜福利一区二区在线看| 啪啪无遮挡十八禁网站| 国产在线精品亚洲第一网站| 一区二区三区高清视频在线| 欧美成人性av电影在线观看| 日韩欧美在线二视频| 91在线观看av| 国产男靠女视频免费网站| 美女高潮到喷水免费观看| 99在线视频只有这里精品首页| 日日夜夜操网爽| 久久人人97超碰香蕉20202| 琪琪午夜伦伦电影理论片6080| 久99久视频精品免费| 久久 成人 亚洲| 又紧又爽又黄一区二区| 美女午夜性视频免费| 黄色丝袜av网址大全| 亚洲一区高清亚洲精品| 欧美一级a爱片免费观看看 | 亚洲国产精品成人综合色| 欧美日韩乱码在线| 如日韩欧美国产精品一区二区三区| 深夜精品福利| 亚洲精品美女久久av网站| videosex国产| 亚洲久久久国产精品| 波多野结衣高清无吗| 国产精品免费一区二区三区在线| cao死你这个sao货| 日本撒尿小便嘘嘘汇集6| 亚洲情色 制服丝袜| av电影中文网址| 亚洲 欧美 日韩 在线 免费| 侵犯人妻中文字幕一二三四区| 亚洲va日本ⅴa欧美va伊人久久| 国产熟女xx| 国产欧美日韩综合在线一区二区| 亚洲七黄色美女视频| 又大又爽又粗| 给我免费播放毛片高清在线观看| 久久久精品国产亚洲av高清涩受| 色尼玛亚洲综合影院| 国产aⅴ精品一区二区三区波| 搞女人的毛片| 色哟哟哟哟哟哟| 亚洲国产精品sss在线观看| 亚洲av熟女| 国产av一区在线观看免费| 亚洲 欧美 日韩 在线 免费| 国产精品,欧美在线| 午夜亚洲福利在线播放| 女警被强在线播放| 亚洲中文日韩欧美视频| 精品久久久久久,| 精品久久久久久久毛片微露脸| 精品国产一区二区久久| 色老头精品视频在线观看| 国产精品久久久人人做人人爽| 成年人黄色毛片网站| 中文字幕高清在线视频| 成人国产一区最新在线观看| 国产精品久久久人人做人人爽| 精品乱码久久久久久99久播| 性欧美人与动物交配| 如日韩欧美国产精品一区二区三区| 大码成人一级视频| 国产成人av激情在线播放| 最近最新中文字幕大全电影3 | 色哟哟哟哟哟哟| 宅男免费午夜| 国产精品美女特级片免费视频播放器 | 男女之事视频高清在线观看| 很黄的视频免费| 又黄又粗又硬又大视频| 国产精品,欧美在线| 99香蕉大伊视频| 女人爽到高潮嗷嗷叫在线视频| 亚洲欧美日韩无卡精品| 亚洲精华国产精华精| 最新美女视频免费是黄的| 大型黄色视频在线免费观看| 欧美成人性av电影在线观看| 在线观看午夜福利视频| 久久精品aⅴ一区二区三区四区| 不卡一级毛片| 午夜免费观看网址| 亚洲狠狠婷婷综合久久图片| 国产精品久久久久久人妻精品电影| 国产精品久久视频播放| 香蕉国产在线看| 欧美日韩福利视频一区二区| 无人区码免费观看不卡| 免费无遮挡裸体视频| 国产亚洲精品综合一区在线观看 | 国产激情久久老熟女| 欧美日韩精品网址| 免费少妇av软件| 日韩高清综合在线| 国产aⅴ精品一区二区三区波| 精品卡一卡二卡四卡免费| 亚洲精品一区av在线观看| 欧美激情高清一区二区三区| 免费不卡黄色视频| 激情在线观看视频在线高清| 国产三级在线视频| 国内久久婷婷六月综合欲色啪| 一级毛片精品| 美女免费视频网站| 免费人成视频x8x8入口观看| 久久久国产精品麻豆| 亚洲一卡2卡3卡4卡5卡精品中文| 日日爽夜夜爽网站| 午夜精品国产一区二区电影| 国产精品av久久久久免费| 成人三级做爰电影| 在线观看免费视频日本深夜| 深夜精品福利| av片东京热男人的天堂| 欧美不卡视频在线免费观看 | 国产成人一区二区三区免费视频网站| 成人18禁在线播放| 黄色毛片三级朝国网站| 欧美在线黄色| bbb黄色大片| 精品国产国语对白av| 久久精品国产亚洲av香蕉五月| 黄色片一级片一级黄色片| 亚洲一区中文字幕在线| 国产一卡二卡三卡精品| bbb黄色大片| 激情在线观看视频在线高清| 国产亚洲欧美在线一区二区| 日本三级黄在线观看| 12—13女人毛片做爰片一| 91成年电影在线观看| 欧美黄色淫秽网站| 中文字幕精品免费在线观看视频| 一级a爱视频在线免费观看| 日韩精品中文字幕看吧| 18禁国产床啪视频网站| 久久精品91无色码中文字幕| 亚洲天堂国产精品一区在线| 99久久久亚洲精品蜜臀av| 1024香蕉在线观看| 久久久久九九精品影院| 亚洲国产精品久久男人天堂| 亚洲国产日韩欧美精品在线观看 | 久9热在线精品视频| 久久青草综合色| 一进一出抽搐gif免费好疼| 欧美黄色片欧美黄色片| 亚洲人成电影观看| 91精品国产国语对白视频| 精品人妻1区二区| 亚洲一码二码三码区别大吗| 欧美成人免费av一区二区三区| 不卡av一区二区三区| 国产三级在线视频| www.www免费av| 搡老熟女国产l中国老女人| 国产91精品成人一区二区三区| 欧美成狂野欧美在线观看| 又黄又粗又硬又大视频| 天堂动漫精品| 日本一区二区免费在线视频| 国产精品一区二区精品视频观看| 美女高潮到喷水免费观看| 日本在线视频免费播放| 免费看a级黄色片| 亚洲五月婷婷丁香| 可以在线观看毛片的网站| www.自偷自拍.com| 亚洲av美国av| 亚洲国产精品999在线| 一本久久中文字幕| 亚洲一区二区三区不卡视频| 国产精品久久久久久精品电影 | 精品少妇一区二区三区视频日本电影| 嫩草影视91久久| 好男人电影高清在线观看| 精品一区二区三区视频在线观看免费| 天堂√8在线中文| 99久久国产精品久久久| 美女国产高潮福利片在线看| 亚洲,欧美精品.| 女人精品久久久久毛片| 亚洲成人免费电影在线观看| 久久中文字幕一级| 亚洲在线自拍视频| 欧美一区二区精品小视频在线| 大型av网站在线播放| 久久国产精品人妻蜜桃| 91老司机精品| 亚洲欧洲精品一区二区精品久久久| 村上凉子中文字幕在线| 操出白浆在线播放| 亚洲午夜精品一区,二区,三区| 久久久国产精品麻豆| 国产人伦9x9x在线观看| 亚洲伊人色综图| 这个男人来自地球电影免费观看| 好看av亚洲va欧美ⅴa在| 国产精品免费一区二区三区在线| 国产精品亚洲av一区麻豆| 成人av一区二区三区在线看| 两个人视频免费观看高清| 啦啦啦韩国在线观看视频| 男人舔女人下体高潮全视频| 亚洲一卡2卡3卡4卡5卡精品中文| 国产精品99久久99久久久不卡| 黄色视频不卡| 欧美成人免费av一区二区三区| 99久久久亚洲精品蜜臀av| 满18在线观看网站| 黄色a级毛片大全视频| 美女午夜性视频免费| 香蕉国产在线看| 亚洲va日本ⅴa欧美va伊人久久| 欧美日本中文国产一区发布| 午夜福利成人在线免费观看| 亚洲精品中文字幕在线视频| 亚洲av日韩精品久久久久久密| av有码第一页| 国产成人av教育| 岛国在线观看网站| 99re在线观看精品视频| 久久久国产成人免费| 动漫黄色视频在线观看| 精品国产乱码久久久久久男人| 国产成人啪精品午夜网站| 亚洲精品国产区一区二| 免费看美女性在线毛片视频| 精品欧美一区二区三区在线| av电影中文网址| 丰满的人妻完整版| 免费在线观看亚洲国产| 香蕉丝袜av| 最新美女视频免费是黄的| 亚洲少妇的诱惑av| 日本 欧美在线| 欧美成人一区二区免费高清观看 | 亚洲国产高清在线一区二区三 | 黄色视频不卡| xxx96com| 99re在线观看精品视频| 免费在线观看亚洲国产| 国产高清视频在线播放一区| 欧美乱妇无乱码| 久久久久国产精品人妻aⅴ院| 亚洲人成77777在线视频| 夜夜躁狠狠躁天天躁| 国产在线观看jvid| 热99re8久久精品国产| 亚洲三区欧美一区| 亚洲欧美一区二区三区黑人| 91精品国产国语对白视频| 日韩欧美三级三区| 亚洲国产中文字幕在线视频| 曰老女人黄片| 少妇 在线观看| 嫩草影院精品99| 国产aⅴ精品一区二区三区波| 嫩草影视91久久| 日韩高清综合在线| 亚洲国产日韩欧美精品在线观看 | 好看av亚洲va欧美ⅴa在| 国产主播在线观看一区二区| 亚洲人成电影免费在线| 97人妻天天添夜夜摸| 午夜亚洲福利在线播放| 亚洲国产欧美网| 免费观看人在逋| 国产一区二区三区综合在线观看| 欧美另类亚洲清纯唯美| 久久国产乱子伦精品免费另类| 女警被强在线播放| 亚洲专区中文字幕在线| 首页视频小说图片口味搜索| av欧美777| 国产精品一区二区在线不卡| 亚洲第一av免费看| 很黄的视频免费| 日本免费一区二区三区高清不卡 | 99久久国产精品久久久| 人妻丰满熟妇av一区二区三区| 好看av亚洲va欧美ⅴa在| 91成年电影在线观看| 国产激情欧美一区二区| 免费av毛片视频| 亚洲国产中文字幕在线视频| 亚洲片人在线观看| 法律面前人人平等表现在哪些方面| 97人妻精品一区二区三区麻豆 | 国产一区二区三区在线臀色熟女| 国产av又大| 少妇的丰满在线观看| 亚洲精品在线观看二区| 搡老妇女老女人老熟妇| 日韩欧美一区视频在线观看| 日韩三级视频一区二区三区| 午夜免费鲁丝| 亚洲熟妇中文字幕五十中出| 国产成人av激情在线播放| 日韩 欧美 亚洲 中文字幕| 精品无人区乱码1区二区| 亚洲精品中文字幕在线视频| 最近最新中文字幕大全免费视频| 亚洲专区字幕在线| 欧美日本中文国产一区发布| 久久久久精品国产欧美久久久| 一区福利在线观看| 可以免费在线观看a视频的电影网站| 日韩精品青青久久久久久| 一进一出抽搐动态| 老司机靠b影院| 18禁黄网站禁片午夜丰满| 亚洲伊人色综图| 国产麻豆69| 成人国产一区最新在线观看| 午夜免费激情av| 麻豆成人av在线观看| 亚洲黑人精品在线| 欧美国产精品va在线观看不卡| 国产成人精品久久二区二区91| 激情在线观看视频在线高清| 亚洲av第一区精品v没综合| 可以在线观看毛片的网站| 国产91精品成人一区二区三区| 首页视频小说图片口味搜索| 村上凉子中文字幕在线| 欧美成人免费av一区二区三区| 欧美av亚洲av综合av国产av| 夜夜夜夜夜久久久久| 大型黄色视频在线免费观看| 国产成人av教育| 亚洲成人久久性| 制服诱惑二区| 夜夜看夜夜爽夜夜摸| 免费女性裸体啪啪无遮挡网站| 在线观看舔阴道视频| 大陆偷拍与自拍| 免费在线观看视频国产中文字幕亚洲| 亚洲av第一区精品v没综合| 啦啦啦观看免费观看视频高清 | 国产精品av久久久久免费| 淫妇啪啪啪对白视频| 日本欧美视频一区| 久久精品成人免费网站| 一边摸一边做爽爽视频免费| 麻豆av在线久日| 老司机靠b影院| 精品一区二区三区四区五区乱码| 久久久久久人人人人人| 国产麻豆69| 91成人精品电影| 99久久综合精品五月天人人| 亚洲国产精品999在线| 欧洲精品卡2卡3卡4卡5卡区| 两个人免费观看高清视频| 国产精品一区二区免费欧美| 色综合亚洲欧美另类图片| 午夜免费鲁丝| 嫩草影院精品99| 国产熟女xx| av视频在线观看入口| 久久草成人影院| 在线观看免费视频网站a站| 多毛熟女@视频| 岛国视频午夜一区免费看| 91老司机精品| 老汉色av国产亚洲站长工具| 精品国内亚洲2022精品成人| 欧美日本中文国产一区发布| 看黄色毛片网站| 国产三级黄色录像| 一区二区三区国产精品乱码| 露出奶头的视频| 日韩精品中文字幕看吧| 午夜免费激情av| 国产乱人伦免费视频| or卡值多少钱| 国产亚洲精品一区二区www| 日本免费a在线| 色老头精品视频在线观看| 琪琪午夜伦伦电影理论片6080| 国产亚洲精品av在线| 两个人视频免费观看高清| 欧美精品啪啪一区二区三区| 免费搜索国产男女视频| 老司机在亚洲福利影院| 高清毛片免费观看视频网站| 999久久久精品免费观看国产| 久久午夜亚洲精品久久| 大型黄色视频在线免费观看| 中文字幕色久视频| 国产私拍福利视频在线观看| 国产99久久九九免费精品| 欧美激情极品国产一区二区三区| 免费在线观看日本一区| 91成年电影在线观看| 美国免费a级毛片| 亚洲av第一区精品v没综合| 精品福利观看| 欧美日韩福利视频一区二区| 村上凉子中文字幕在线| 大码成人一级视频| 免费av毛片视频| 亚洲欧洲精品一区二区精品久久久| 国产精品久久久久久精品电影 | 在线播放国产精品三级| 国产欧美日韩一区二区三| 久久久久久久久免费视频了| 美女高潮到喷水免费观看| 在线观看舔阴道视频| 欧美黑人欧美精品刺激| 久久久精品欧美日韩精品| 91九色精品人成在线观看| 黄色视频不卡| 免费观看精品视频网站| 国产精品一区二区三区四区久久 | 久久久精品欧美日韩精品| 亚洲最大成人中文| 一个人观看的视频www高清免费观看 | 亚洲av第一区精品v没综合| 亚洲精品一卡2卡三卡4卡5卡| 真人一进一出gif抽搐免费| 久久久久久人人人人人| 国产亚洲av嫩草精品影院| 12—13女人毛片做爰片一| 国产精品秋霞免费鲁丝片| av天堂在线播放| 一卡2卡三卡四卡精品乱码亚洲| 国产精品,欧美在线| 少妇熟女aⅴ在线视频| 午夜老司机福利片| 美女高潮喷水抽搐中文字幕| 韩国av一区二区三区四区| 夜夜爽天天搞| 亚洲国产高清在线一区二区三 | 淫秽高清视频在线观看| 韩国av一区二区三区四区| 国产一区二区三区在线臀色熟女| 午夜福利影视在线免费观看| 麻豆av在线久日| 国产高清视频在线播放一区| 中文字幕色久视频| 国产私拍福利视频在线观看| 一级毛片精品| 99国产综合亚洲精品| 亚洲av五月六月丁香网|