• <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 ?

    Efficacy on rabbits with arrhythmia:needling acupoint of Neiguan(PC6)at shallow or deep depth,and retaining needles for 10,20,or 30 min

    2021-08-09 12:36:44GUChunlingCHENGKaiYANGShouqinRANShaofengTAIHengyapGAOXiaofengPANGDandan

    GU Chunling,CHENG Kai,YANG Shouqin,RAN Shaofeng,TAI Hengyap,GAO Xiaofeng,PANG Dandan

    GU Chunling,YANG Shouqin,RAN Shaofeng,TAI Hengyap,School of Acupuncture and Tuina,Beijing University of Chinese Medicine,Beijing 100029,China

    CHENG Kai,School of Acupuncture and Tuina,Beijing University of Chinese Medicine,Beijing 100029,China

    GAO Xiaofeng,Beijing MedEx Technology Co.,Ltd.,Beijing 100029,China

    PANG Dandan,China Academy of Chinese Medical Sciences,Beijing 100700,China

    Abstract OBJECTIVE:To compare the effects of Neiguan(PC6) acupuncture at different depths and retention time on arrhythmia duration,myocardial tissue morphology,mRNA expression level of L-type calcium channel α1C subunit and Ca2+-Mg2+-AtPase activity in tachirrhythmia model of rabbits.METHODS:The tachyarrhythmia model was made by intravenous injection of barium chloride into the ears of rabbits.A total of 56 healthy adult male New Zealand big-eared white rabbits,apply the random number table method,divided into normal control group (group A),model group (group B),shallow needling Neiguan (PC6) 10 min group(group C),shallow needling Neiguan (PC6) 20 min group (group D),shallow needling Neiguan (PC6)30 min group (group E),deep needling Neiguan(PC6) 10 min group (group F),deep needling Neiguan(PC6)20 min group(group G),deep needling Neiguan(PC6)30 min group(group H),7 animals in each group.Electrocardiograms were used to collect the duration of arrhythmia;hematoxylin-eosin staining method was performed on myocardial tissue,RT-PCR tested the expression of α1C subunit mRNA,and the activity of Ca2+-Mg2+-ATPase were quantified by phosphorus determination method.RESULTS:The duration of arrhythmia in each acupuncture treatment group was shortened to varying degrees.Compare to the model group,the tissue damage from barium chloride inducing was improved in the acupuncture group.Compared to the model group,except for group E,most treatment groups had varying degrees of improvement with significantly down-regulated L-type calcium channel α1C subunit mRNA expressions level and increased Ca2+-Mg2+-ATPase activity.CONCLUSIONS:The effect of acupuncture at Neiguan (PC6) with different depths and retention time can reduce the duration of arrhythmia induced by barium chloride relatively,improve the induced pathological changes,down regulate L-type calcium channel α1C subunit mRNA expressions level and increase Ca2+-Mg2+-ATPase activity.Both the shallow and deep tissues of Neiguan(PC6)may be involved in transmitting acupuncture information.There is an optimal induction period for shallow needling at Neiguan (PC6) to reach the best therapeutic effect.

    Keywords:Point PC6(Neiguan);arrhythmias,cardiac;calcium channels,L-type;Ca2+-Mg2+-ATPase;needle retention time

    INTRODUCTION

    Neiguan (PC6) is the first choice for acupuncture and moxibustion regarding arrhythmia treatment.1There are kinds of acupuncture levels methods,such as 1/5 inch,half-inch,one inch,and penetration acupuncture through with Waiguan (SJ5).Studies have confirmed that both shallow needling (3 mm) and deep needling(5-8 mm) at Neiguan (PC6) can treat arrhythmias.2The specificity of acupoints reflects in different regulating effects of acupuncture at different levels and retention times to reach its best effect.3Most clinical studies on the relationship of the acupuncture's effect between different depths have shown that deep needling was better than shallow needling.4Previous studies proved that deep needling at Neiguan (PC6) was better than shallow needling in shortening the duration of atrial fibrillation.5Does the poor efficacy of shallow needling have something to do with the needle retention time?The needle retention time is too short to achieve the best therapeutic effect?Or is it too long that may cause tolerance and fatigue,resulting in ineffective or even inferior stimulation?6

    The purpose of this experiment is to explore the acupuncture level and needle retention time to obtain the best therapeutic effect of tachyarrhythmia,in order to provide scientific basis for clinical rational use of acupoints.

    MATERIALS AND METHODS

    Experimental animals

    A total of 56 healthy and clean adult male New Zealand white rabbits,weighing 2.0-2.5 kg.Provided by Beijing Fulong Tengfei Breeding Center,license number:SCXK (Peking) 2018-0009.They are raised in the animal room on the Liangxiang campus of Beijing University of TCM.The breeding conditions are:indoor temperature (18-22) ℃,humidity 35%-45%,natural light,food and water are provided.Rabbits were fasting 12 h before the experiment operation,shaver were used to remove the rabbit hair on the inner side of the both forearm in order to accurately locate the acupuncture points.The study was approved by the experimental animal ethics committee of Beijing University of TCM.

    Experiment grouping

    A total of 56 rabbits applies the random number table method dividing into eight groups,seven animals in each one.There were named with the control group(group A)and the model group(group B),shallow needling Neiguan (PC6) 10 min group (Group C),shallow needling Neiguan (PC6) 20 min group (group D),shallow needling Neiguan (PC6) 30 min group (group E),deep needling Neiguan (PC6) 10 min group(group F),deep needling Neiguan (PC6) 20 min group (group G),deep needling Neiguan (PC6)30 min group(group H).

    Experimental reagents and instruments

    Chloral hydrate was purchased from Honghu United Chemical Industry (Beijing,China);Anhydrous Barium Chloride was purchased from Yinuokai Biology Company (Beijing,China);HiFiScript cDNA first-strand synthesis kit was purchased from Cwbiotech (Taizhou,China);SYBR FAST qPCR Kit Master Mix (2×) Universal was purchased from American KAPA Biosystems (Woburn,MA,USA);Trizol was purchased from Invitrogen (Carlsbad,California,USA);Chloroform,isopropanol was purchased from Beijing Chemical Reagent Factory (Beijing,China);Ultramicro ATP enzyme(Ca2+-Mg2+-ATP) assay kit and Total Protein assay were purchased from Beijing Rigor Bioscience(Beijing,China).

    BL-420N biological signal acquisition and analysis system was purchased from Chengdutaimeng Biology Company (Chengdu,Sichuan,China);disposable sterile acupuncture needles was purchased from Zhongyantaihe Company (Beijing,China);spectrophotometer:UV-2000 was purchased from Jinghua Technology Instrument Company (Shanghai,China);real-time quantitative PCR instrument:ABI7500 was purchased from Applied Biosystems (Foster,CA,USA);fully automatic biochemical analyzer:UniCel DxC 600 Synchron was purchased from Beckman Coulter (Brea,CA,USA);Gel imager:BioSens SC 810B was purchased from Shanghai Shanfu Biotech Company(Shanghai,China).

    Modeling method

    After weighing,the rabbits were anesthetized by intraperitoneal injection of chloral hydrate (10%,5 mL/kg).7After completing anesthesia,the rabbit was fixed on the rabbit table in the supine position.Disposable sterile needles were used as electrodes and inserted under rabbits' skin at the right forelimb and the ends of both lower limbs.After connecting the BL-420N bio-signal acquisition and analysis system ECG electrodes to the rabbit,run the standard limb II electrocardiogram (ECG) and monitor the ECG for 5 min.In the given min,a bolus injection through the ear vein with sodium chloride solution (0.9%,1 mL/kg) was given to the control group;the other groups were injected with barium chloride solution (0.4%,1mL/kg)through the ear vein.8A successful model will be verified with the occurrence of 7 or more continuous premature ventricular contraction beats (PVC),9the PVC goes by the definition of continuous,vast,deformed,or torsion QRS waveform;with not preceded P waves and unequal R waves gap.The rate of change in heart rate after barium chloride injection versus before was greater than 0.39.

    Acupuncture method

    Refer to 10to locate the Neiguan (PC6) in rabbits:The lower 1/6 point of the inner forearm (approximately 1.2-1.3 cm above the wrist joint),between the flexor carpi radialis and the flexor digitorum superficialis.After the model is successfully build,prepare a disposable sterile acupuncture needle of 0.25 mm × 25 mm.Process the shallow acupuncture group needling straightly with a depth of 3 mm2the tip of the needle stops at the surface of the muscle,retain the needle for 10 min(group C),20 min (group D),and 30 min (group E).In like manner,process the deep acupuncture group needling straightly with a depth of 5-8 mm,11,12properly adjust the direction of the needle tip to touch the endings of median nerve to make the rabbit limbs twitch,retain the needles for 10 min (group F),20 min(group G),and 30 min(group H).Each group were performed with the tremorQistimulation method (that is,hold the needle handle and do a small and rapid tremor without relative displacement between the needle body and the surrounding tissues) once every 3 min with a 3 min length,this was repeated until the needle retention time was over.Group A and group B were not performed with acupuncture.

    Observation indicators and detection methods

    (a) Arrhythmia duration:the ECG was recorded continuously with limb lead-II standard until the sinus rhythm was restored during the experiment.The arrhythmia duration is counted from the first ventricular premature contraction beat on ECG to the beginning of the recovery of sinus rhythm.13Ventricular tachycardia disappears for more than 300 s can be diagnosed as the restoration of the rather normal heart rhythm.14

    (b) Observation of myocardial morphologyviahematoxylin-eosin staining method:rabbit's left ventricular anterior wall myocardial tissue specimen was fixed in 4% paraformaldehyde,then dehydrated by gradient ethanol,transparent xylene,immersed in wax,embedded,and sectioned.The thickness of the section was about 5 μm,hematoxylin-eosin staining was performed,and the sections were sealed and observed under a microscope.

    (c) RT-PCR technique was used to observe the mRNA expression level of LTCC α1C subunit:extract total RNA was extracted from the tissue with TRIzol reagent,reverse transcription kit was used to synthesize cDNA by reverse transcription.The primers and cDNA were added to the SYBR FAST qPCR Kit Master Mix (2×) Universal PCR system,respectively,then the experiment was performed in the real-time fluorescence quantitative PCR (plus stepone,ABI),with a final volume of 20 μL.GAPDH was used for references,and the 2-△△Ctmethod was used for relative quantitative analysis of the data.All experiments were repeated at least three times(Tables 1,2).

    Table 1 Primer sequence of detection gene

    Table 2 Reaction conditions

    (d) Determination of Ca2+-Mg2+-ATPase activity via phosphorus determination method:After collecting rabbit's cardiac muscle tissue,prepare normal saline tissue homogenate,centrifuge at 3000-4000 r/min for 10 min,and then extract 100 μL of supernatant.After enzymatic reaction,comparison tube and measuring tube,extracted the supernatant 300 μL phosphonium,Ca2+-Mg2+-ATPase activity value detection,according to the kit instructions,enzyme activity was determined by the enzymatic reaction and phospho-assay method.

    Statistical analysis

    All data were analyzed using SAS (vers 9.4,SAS Institute,Chicago,IL,USA) software.Measurement data were expressed as mean ± standard deviation (±s).When comparing between the groups,the data of each group was conformed to normality and homogeneity of variance using two independent sample t-tests or analysis of variance.There were statistical differences in the analysis of variance,and the least significant difference test was used for pairwise comparison between groups.The interaction was analyzed by factorial (2*3)analysis designed of variance.P <0.05 was used as the standard for statistically significant differences.

    RESULTS

    Comparison of intervention effects on the duration of tachyarrhythmia in rabbits of each group

    Compared with the model group [(4387 ± 728) s],the duration of arrhythmia in each acupuncture treatment group was shortened to varying degrees (P <0.05).In terms of the duration of arrhythmia,shallow acupuncture groups showed that group D [(1636 ± 196) s]

    Comparison of the morphology of rabbit ventricular myocytes in each group

    In Figure 1,the rabbit's left ventricle anterior wall of the cardiac muscle was observed.The cardiac muscle tissue structure of the control group was typical;the cell nucleus was intact and elliptical with narrow blankspace surrounding it;both longitudinal and transverse sections showing that the cytoplasm was relatively uniform and had no abnormal changes.The ventricular wall fibers were densely arranged near the center in sheets-like formations;the tissue near the heart cavity(papillary muscles)was loose and in fascicle order.

    Figure 1 Hematoxylin-eosin staining results of rabbit myocardium(cross section,×200 times)

    In the model group,there were noticeable structural changes:pronounced edema of the cardiomyocytes forming it into granular shape,vacuoles changes in myocardial cells;some of the nuclei have a sign of lysis,shrinkage and wrinkled around the edge forming into irregular shapes;widened perinuclear space;pale stained cytoplasm;disordered fiber arrangement and distinctive wave-like degeneration.

    In the control group and model groups,capillaries could be seen between the myocardial cells.There were different degrees of myocardial cell edema,cell vacuolation,myocardial fiber arrangement disorder,wrinkled nucleus,widened perinuclear space,pale stained cytoplasm,and wave-like degeneration in each acupuncture treatment group.Capillary dilation with new red liquid in between was seen in all acupuncture treatment groups.

    Comparison mRNA expression of L-type calcium channel α1C subunit in myocardial tissue of rabbits in different groups

    Compared with the control group (0.25 ± 0.04),the L-type calcium channel α1C subunit mRNA expression of the model group (1.49 ± 0.29) is significantly higher (P <0.01).The expression of majority treatment group noticeably decreased(P<0.05),except for group E (P >0.05).Comparing the expression level of each group showing a result of — shallow acupuncture groups,group D (0.65 ± 0.25)

    Comparison Ca2+-Mg2+-ATPase activity in myocardial tissue of rabbits in various groups

    Compared with the control group [(8.4 ± 0.6) μmol·mg-1·h-1],the activity of Ca2+-Mg2+-ATPase was decreased in the model group[(7.3±0.5)μmol·mg-1·h-1]and the difference was statistically significant (P <0.01).Except for the group E (P >0.05),compared with the model group,the activities of Ca2+-Mg2+-ATPase in other acupuncture treatment groups were increased in varying degrees,and the difference was statistically significant(P<0.05).In the activity of Ca2+-Mg2+-ATPase,shallow acupuncture groups showed that group E [(7.3 ± 0.4) μmol·mg-1·h-1]

    DISCUSSION

    The modeling method in this experiment is to inject barium chloride solutionviathe ear vein to induce tachyarrhythmia in rabbits.After the induction,the incidence rate of ventricular tachycardia is 100%,as well as the survival rate,proving that the induction method is successful.The sinus rhythm of the model group's rabbit usually needs 60-70 min to recover after the induction of barium chloride,but in comparison to the model group,each acupuncture treatment group it shortened the duration of arrhythmia to varying degrees.

    Compared to the control group,the tissue from the model group,which was damaged by the injection of barium chloride,shows noticeable structural changes.Damage such as nucleus shrinkage,lysis,disordered fiber arrangement,and substantial wave-like degeneration were seen widely.On the other hand,the pathological changes in the treatment group were lessen to varying degrees,dilated capillaries filled with red liquid were also seen.We speculate that acupuncture treatments may increase the myocardial blood flow,which triggers this phenomenon.For groups G and H,the myocardial morphology tends to be normal,reflecting that acupuncture Neiguan (PC6) has a certain degree of therapeutic protection against myocardial damage.

    Mechanism of arrhythmia induced by barium chloride may be related to delayed depolarization and triggering activity induced by increased Na+/Ca2+influx in myocardial working cells.15,16In the plateau phase of the myocardial action potential,extracellular Ca2+enters into the cell through the L-type calcium ion channel,which increased the intracytoplasmic Ca2+concentration and triggered the release of large amounts of Ca2+from the sarcoplasmic reticulum.17-19Ca2+-Mg2+-ATPase can pump a small amount of Ca2+out of the cells or into the sarcoplasmic reticulum when the intracellular Ca2+was increased to a certain extent,thereby reducing the intracellular Ca2+concentration.20

    The density and distribution of LTCC joined a crucial role in the occurrence and maintenance of arrhythmia,21,22α1C is the main subunit that constitutes in ventricular L-type calcium channels,abnormal changes of α1C protein expression at right ventricular outflow tract trigger arrhythmia;23,24the inactivation of α1C and down-regulated mRNA expression promoted electrical remodeling of cardiomyocytes which induct and maintain the event of arrhythmia.25-27Comparing with the control group,the expression of LTCC was up-regulated in the inducted groups,resonating with the cited article above.LTCC expression in other acupuncture treatment groups excluding group E decreased to varying degrees compared with the model group.It shows that acupuncture of different depth at Neiguan (PC6) had a certain degree of effectiveness of down regulating the expression of α1C mRNA.This result is substantiated with a previous study — Neiguan(PC6)regulatory effect on the expression of calcium ion transmembrane channels in cardiomyocytes.28Decreased Ca2+-Mg2+-ATPase activity will cause intracellular calcium overload,damage mitochondria's function,increase the concentration of oxygen radicals,and therefore causes arrhythmia.29-31Compared to the model group with the control group,Ca2+-Mg2+-ATPase activity decreased after the successful modeling in this experiment,which substantiates the theory of the cited article above.Furthermore,compared to the model group,except for group E;other treatment groups had various increases in Ca2+-Mg2+-ATPase activity,indicating that acupuncture at Neiguan (PC6) can increase the activity of Ca2+-Mg2+-ATPase,consequently reduces the overload of intracellular calcium and the degree of cardiomyocytes' damage,showing a corresponding result with the cited article.32

    In this experiment,the statistical results of each index proved that the curative effects of acupuncture comparison for both the deep needling and shallow needling at Neiguan (PC6) were statistically significant (P <0.05).Thus,it suggested that both the shallow and deep tissues of Neiguan (PC6) may participate in the transmission of acupuncture information.

    The effect trend of each index showed that the effect of superficial level's curative from large to small was acupunctured 20,10,30 min.The therapeutic effect of shallow needling for 20 min was better than that of shallow needling for 30 min (P <0.05),which suggest that there are an optimal induction period for shallow needling at Neiguan(PC6).

    The degree of deep needling curative effect ranges from large to small was acupunctured 30,20,and 10 min.With the prolongation of the needle retaining time,the acupuncture effect of deep needling at Neiguan (PC6) was gradually enhanced,but whether the acupuncture effect will continue strengthening or gradually decreased or produces negative feedback with the further prolongation needle retaining time remains to be proved by further research.

    The stimulation effect of shallow needling was more sensitive to the response of needle retention time,the optimal induction period was shorter,and it was easier to produce invalid stimulation due to long needle retention time,suggesting that shallow needling should be paying more attention to the relationship of time and effect.

    日韩一本色道免费dvd| 亚洲精品aⅴ在线观看| 国产伦精品一区二区三区四那| 国产 一区 欧美 日韩| 亚洲av不卡在线观看| 超碰av人人做人人爽久久| 日韩大片免费观看网站| 亚洲人成网站在线观看播放| 自拍欧美九色日韩亚洲蝌蚪91 | 精品久久久久久久久av| av免费观看日本| 亚洲精品自拍成人| 国产老妇女一区| 女人久久www免费人成看片| 国产大屁股一区二区在线视频| 国产精品国产三级国产专区5o| 欧美成人a在线观看| 97人妻精品一区二区三区麻豆| 成人亚洲欧美一区二区av| 国产毛片a区久久久久| 日日摸夜夜添夜夜添av毛片| 一级爰片在线观看| 韩国av在线不卡| 国产黄色视频一区二区在线观看| 狂野欧美激情性xxxx在线观看| 亚洲国产欧美在线一区| 国产成人精品婷婷| 欧美日韩精品成人综合77777| 人妻一区二区av| 亚洲国产成人一精品久久久| 天堂俺去俺来也www色官网| 深爱激情五月婷婷| 久久ye,这里只有精品| 久久久国产一区二区| 午夜日本视频在线| 日本-黄色视频高清免费观看| 在线天堂最新版资源| 成人亚洲精品一区在线观看 | 日韩av免费高清视频| 在线看a的网站| 岛国毛片在线播放| 久久亚洲国产成人精品v| 国产午夜精品一二区理论片| 新久久久久国产一级毛片| 国产精品伦人一区二区| 在线亚洲精品国产二区图片欧美 | 狂野欧美白嫩少妇大欣赏| av国产久精品久网站免费入址| 亚洲内射少妇av| 国产成人免费观看mmmm| 国产精品一及| 欧美日韩视频精品一区| 2021天堂中文幕一二区在线观| 亚洲精品久久久久久婷婷小说| 人妻制服诱惑在线中文字幕| 人人妻人人看人人澡| 成人美女网站在线观看视频| av在线老鸭窝| 好男人在线观看高清免费视频| 99久久精品国产国产毛片| 国产片特级美女逼逼视频| 久久精品综合一区二区三区| 国产一区二区在线观看日韩| 欧美日韩在线观看h| 亚洲自拍偷在线| 国产亚洲5aaaaa淫片| 成人毛片60女人毛片免费| av在线蜜桃| 午夜福利网站1000一区二区三区| 人人妻人人澡人人爽人人夜夜| 国产成人一区二区在线| 女人被狂操c到高潮| 中文乱码字字幕精品一区二区三区| 国产免费视频播放在线视频| 午夜亚洲福利在线播放| 91在线精品国自产拍蜜月| 少妇丰满av| 国产精品久久久久久精品电影| 身体一侧抽搐| 看十八女毛片水多多多| 三级男女做爰猛烈吃奶摸视频| 久久精品国产亚洲av涩爱| 国产精品99久久久久久久久| 欧美人与善性xxx| 内射极品少妇av片p| 熟女av电影| 国产午夜精品一二区理论片| 国产精品人妻久久久久久| 亚洲三级黄色毛片| 人妻 亚洲 视频| 午夜视频国产福利| 看免费成人av毛片| 亚洲精品,欧美精品| 日韩强制内射视频| 日韩亚洲欧美综合| 亚洲美女搞黄在线观看| 国产精品99久久久久久久久| 国产高清不卡午夜福利| 伊人久久精品亚洲午夜| 我的老师免费观看完整版| 免费av毛片视频| 午夜亚洲福利在线播放| 国产精品99久久99久久久不卡 | 久久午夜福利片| 久久久久九九精品影院| 中文字幕亚洲精品专区| 秋霞伦理黄片| 欧美三级亚洲精品| 在线观看一区二区三区| 日韩强制内射视频| 亚洲人与动物交配视频| 成人美女网站在线观看视频| 免费av不卡在线播放| 婷婷色麻豆天堂久久| 免费不卡的大黄色大毛片视频在线观看| 日韩成人av中文字幕在线观看| 日韩亚洲欧美综合| 日本三级黄在线观看| av在线亚洲专区| 在线看a的网站| 欧美bdsm另类| 性色avwww在线观看| 国产精品麻豆人妻色哟哟久久| 亚洲av欧美aⅴ国产| 嫩草影院精品99| 美女视频免费永久观看网站| 九九在线视频观看精品| 亚洲av免费在线观看| 在线免费十八禁| 日韩在线高清观看一区二区三区| 搡女人真爽免费视频火全软件| 日本一二三区视频观看| 尾随美女入室| 亚洲综合色惰| 亚洲精品成人av观看孕妇| 久久精品国产亚洲av涩爱| 日本wwww免费看| 毛片女人毛片| 欧美老熟妇乱子伦牲交| 搡女人真爽免费视频火全软件| 毛片女人毛片| 久久久欧美国产精品| 哪个播放器可以免费观看大片| 一区二区三区免费毛片| 看黄色毛片网站| 熟女av电影| 免费看光身美女| 日本wwww免费看| 亚洲精品日本国产第一区| 国产精品国产av在线观看| 日本午夜av视频| 18禁在线无遮挡免费观看视频| 亚洲三级黄色毛片| 哪个播放器可以免费观看大片| 精品久久久精品久久久| 亚洲精品一二三| 精品国产乱码久久久久久小说| 最近手机中文字幕大全| 小蜜桃在线观看免费完整版高清| 又大又黄又爽视频免费| 色婷婷久久久亚洲欧美| 欧美人与善性xxx| 亚洲av男天堂| 麻豆乱淫一区二区| 99精国产麻豆久久婷婷| 国产精品精品国产色婷婷| 亚洲不卡免费看| 两个人的视频大全免费| 久久6这里有精品| 老女人水多毛片| 日韩一本色道免费dvd| 亚洲色图综合在线观看| 黄色欧美视频在线观看| 精华霜和精华液先用哪个| 最后的刺客免费高清国语| 成人午夜精彩视频在线观看| 久久99热这里只有精品18| 国产精品一区www在线观看| 在线精品无人区一区二区三 | 18禁在线播放成人免费| 国产在线一区二区三区精| 国产高潮美女av| 免费观看性生交大片5| tube8黄色片| 婷婷色麻豆天堂久久| 性插视频无遮挡在线免费观看| 精品一区二区三卡| 精品久久久久久久久亚洲| 精品久久久精品久久久| 亚洲欧美成人精品一区二区| 99久久中文字幕三级久久日本| 欧美xxxx性猛交bbbb| 五月开心婷婷网| 一级片'在线观看视频| 亚洲精品456在线播放app| 欧美日韩综合久久久久久| a级毛片免费高清观看在线播放| 成年av动漫网址| 1000部很黄的大片| 久久97久久精品| 国产一区有黄有色的免费视频| 欧美亚洲 丝袜 人妻 在线| 国模一区二区三区四区视频| 80岁老熟妇乱子伦牲交| 九九久久精品国产亚洲av麻豆| 国产 精品1| 九草在线视频观看| 久久精品综合一区二区三区| 在线观看人妻少妇| eeuss影院久久| 久久久色成人| 国产欧美亚洲国产| 久久ye,这里只有精品| 尤物成人国产欧美一区二区三区| 国产极品天堂在线| 男插女下体视频免费在线播放| 人人妻人人爽人人添夜夜欢视频 | 日本免费在线观看一区| 亚洲av电影在线观看一区二区三区 | 亚洲色图av天堂| 国产黄a三级三级三级人| 欧美日韩亚洲高清精品| 国产黄色免费在线视频| 干丝袜人妻中文字幕| 人人妻人人澡人人爽人人夜夜| 亚洲内射少妇av| 中文字幕免费在线视频6| 国产爱豆传媒在线观看| 日本一二三区视频观看| 三级经典国产精品| 欧美日韩视频高清一区二区三区二| videos熟女内射| 一区二区三区精品91| 高清欧美精品videossex| 搞女人的毛片| 日韩强制内射视频| 精品视频人人做人人爽| 亚洲国产精品专区欧美| 中文字幕亚洲精品专区| av播播在线观看一区| 日韩亚洲欧美综合| 最近中文字幕2019免费版| 欧美一区二区亚洲| 青春草视频在线免费观看| 久久精品久久久久久噜噜老黄| 久久精品夜色国产| av在线蜜桃| 国产精品久久久久久av不卡| 日韩国内少妇激情av| 亚洲欧洲日产国产| 国产色爽女视频免费观看| 欧美日韩视频精品一区| 99热全是精品| 午夜福利高清视频| 好男人在线观看高清免费视频| 看十八女毛片水多多多| 一级二级三级毛片免费看| 国产精品麻豆人妻色哟哟久久| 黄片wwwwww| 国产成人aa在线观看| 国产精品一区二区在线观看99| 久久久精品欧美日韩精品| 亚洲天堂国产精品一区在线| 又大又黄又爽视频免费| 国产精品久久久久久久久免| 亚洲一区二区三区欧美精品 | 国内少妇人妻偷人精品xxx网站| 天天躁日日操中文字幕| 人妻一区二区av| 国产黄a三级三级三级人| 又爽又黄a免费视频| 国产一区二区三区av在线| 丝袜脚勾引网站| 久久久久久久久久人人人人人人| 亚洲,一卡二卡三卡| 亚洲人成网站在线观看播放| 亚洲欧洲日产国产| 日韩电影二区| 午夜激情久久久久久久| 亚洲美女搞黄在线观看| 精品国产乱码久久久久久小说| 少妇熟女欧美另类| 日韩av在线免费看完整版不卡| 18禁裸乳无遮挡免费网站照片| 日韩,欧美,国产一区二区三区| 国产一区二区亚洲精品在线观看| 免费少妇av软件| 精品一区二区免费观看| 别揉我奶头 嗯啊视频| 国产黄频视频在线观看| 日韩一区二区三区影片| 国产极品天堂在线| 秋霞在线观看毛片| 亚洲av中文av极速乱| 国产亚洲精品久久久com| 一个人看的www免费观看视频| 成人漫画全彩无遮挡| 日韩一区二区视频免费看| 丰满人妻一区二区三区视频av| 久久久久久久大尺度免费视频| 亚洲精品,欧美精品| 人妻少妇偷人精品九色| 男的添女的下面高潮视频| 高清欧美精品videossex| 久久久久久久久久久免费av| 丝袜美腿在线中文| 99re6热这里在线精品视频| 激情五月婷婷亚洲| 赤兔流量卡办理| 国产av不卡久久| 国产 一区 欧美 日韩| 亚洲av成人精品一二三区| 亚洲精品中文字幕在线视频 | 97超碰精品成人国产| 欧美精品国产亚洲| 一级爰片在线观看| 亚洲av福利一区| 中文字幕人妻熟人妻熟丝袜美| 26uuu在线亚洲综合色| 成人鲁丝片一二三区免费| 久久精品人妻少妇| 国产精品久久久久久精品电影小说 | 中文字幕av成人在线电影| 最近手机中文字幕大全| 精品熟女少妇av免费看| 高清欧美精品videossex| 老司机影院毛片| 卡戴珊不雅视频在线播放| 亚洲最大成人中文| 亚洲国产精品成人综合色| 天天一区二区日本电影三级| 一区二区三区四区激情视频| 色综合色国产| 卡戴珊不雅视频在线播放| 精品少妇黑人巨大在线播放| 最近中文字幕高清免费大全6| 国产成人一区二区在线| 久久久久久久精品精品| 日韩国内少妇激情av| 免费大片18禁| 国产伦在线观看视频一区| 青春草亚洲视频在线观看| 99久久精品一区二区三区| 一边亲一边摸免费视频| 成年女人在线观看亚洲视频 | 久久久久久久久久成人| 中国国产av一级| 97超碰精品成人国产| 亚洲精品国产成人久久av| 免费人成在线观看视频色| 久久人人爽人人爽人人片va| 日韩欧美一区视频在线观看 | 高清日韩中文字幕在线| 国产综合懂色| 777米奇影视久久| 一级爰片在线观看| 国产黄a三级三级三级人| 精品视频人人做人人爽| 丰满乱子伦码专区| 日韩成人伦理影院| 一级毛片久久久久久久久女| 亚洲最大成人中文| 欧美成人a在线观看| 人人妻人人澡人人爽人人夜夜| 欧美成人a在线观看| 国产黄片美女视频| 久久久久九九精品影院| 大话2 男鬼变身卡| 日韩 亚洲 欧美在线| 日韩一本色道免费dvd| 亚洲人与动物交配视频| 免费看a级黄色片| 三级国产精品片| 国产精品偷伦视频观看了| 韩国高清视频一区二区三区| 国产一区二区三区av在线| 免费看日本二区| 精华霜和精华液先用哪个| 女的被弄到高潮叫床怎么办| 国产精品秋霞免费鲁丝片| 国产在线一区二区三区精| 免费av毛片视频| 在线观看人妻少妇| 九色成人免费人妻av| 精品视频人人做人人爽| 国产高清国产精品国产三级 | 黄片无遮挡物在线观看| 极品教师在线视频| 日本wwww免费看| 极品少妇高潮喷水抽搐| 黑人高潮一二区| 中文字幕久久专区| 噜噜噜噜噜久久久久久91| 熟女av电影| 午夜亚洲福利在线播放| 三级经典国产精品| 国产亚洲av嫩草精品影院| 天堂中文最新版在线下载 | 精品国产一区二区三区久久久樱花 | 在线观看美女被高潮喷水网站| av在线亚洲专区| 日韩一本色道免费dvd| 少妇 在线观看| 只有这里有精品99| 国产欧美日韩一区二区三区在线 | 久久久精品免费免费高清| 91久久精品国产一区二区三区| 亚洲人成网站在线观看播放| 天美传媒精品一区二区| 波野结衣二区三区在线| 男人和女人高潮做爰伦理| 国内少妇人妻偷人精品xxx网站| 在线免费观看不下载黄p国产| 2022亚洲国产成人精品| 国产精品久久久久久久久免| 美女国产视频在线观看| 欧美一级a爱片免费观看看| 国产一区亚洲一区在线观看| 男人添女人高潮全过程视频| 久久ye,这里只有精品| 亚洲,欧美,日韩| 久久97久久精品| av天堂中文字幕网| 亚洲av免费高清在线观看| 亚洲av欧美aⅴ国产| 99热全是精品| 少妇丰满av| 国产日韩欧美亚洲二区| 亚洲内射少妇av| 亚洲精品一二三| 最近中文字幕高清免费大全6| 网址你懂的国产日韩在线| 久久久久九九精品影院| 日韩av不卡免费在线播放| 人妻系列 视频| 久久99精品国语久久久| 久久久久久久大尺度免费视频| 99久久精品热视频| 国产淫语在线视频| 超碰av人人做人人爽久久| 国产成人精品福利久久| 亚洲综合精品二区| 亚洲精品亚洲一区二区| 亚洲国产成人一精品久久久| 国产精品国产三级国产av玫瑰| 亚洲丝袜综合中文字幕| 亚洲av一区综合| 少妇高潮的动态图| 男人和女人高潮做爰伦理| 午夜日本视频在线| 国产欧美日韩一区二区三区在线 | 尾随美女入室| 大片免费播放器 马上看| 国产免费福利视频在线观看| 三级国产精品片| 伦理电影大哥的女人| 大片免费播放器 马上看| 欧美xxⅹ黑人| a级毛色黄片| 久久精品久久久久久噜噜老黄| 中文字幕人妻熟人妻熟丝袜美| 国产av不卡久久| 亚洲无线观看免费| 精品久久久久久久末码| 两个人的视频大全免费| 超碰97精品在线观看| 少妇人妻 视频| 一级爰片在线观看| 18禁在线播放成人免费| 色网站视频免费| 人妻少妇偷人精品九色| 精华霜和精华液先用哪个| videos熟女内射| 97在线视频观看| 在线精品无人区一区二区三 | av在线老鸭窝| 舔av片在线| 久久久久久久久久久免费av| 亚洲国产精品成人久久小说| 少妇人妻一区二区三区视频| 国产色爽女视频免费观看| 亚洲精品国产色婷婷电影| 大香蕉久久网| 国产精品嫩草影院av在线观看| 久久99精品国语久久久| 日日啪夜夜爽| 亚洲国产欧美人成| 99久久九九国产精品国产免费| 国内精品宾馆在线| 91aial.com中文字幕在线观看| 亚洲av在线观看美女高潮| 一边亲一边摸免费视频| 亚洲欧美日韩东京热| 国语对白做爰xxxⅹ性视频网站| 成人毛片60女人毛片免费| 超碰97精品在线观看| 一级毛片 在线播放| 国产精品国产av在线观看| 色播亚洲综合网| 亚洲精品aⅴ在线观看| 亚洲精品中文字幕在线视频 | 寂寞人妻少妇视频99o| 国产视频内射| 亚洲国产精品专区欧美| 午夜免费男女啪啪视频观看| 亚洲欧美日韩卡通动漫| 午夜福利在线观看免费完整高清在| 少妇人妻精品综合一区二区| 国产精品av视频在线免费观看| 一级毛片aaaaaa免费看小| 久久精品久久精品一区二区三区| 寂寞人妻少妇视频99o| 干丝袜人妻中文字幕| 成人国产av品久久久| 美女xxoo啪啪120秒动态图| 黄色日韩在线| 亚洲成人久久爱视频| 久久精品久久久久久噜噜老黄| 青春草视频在线免费观看| 久久99热这里只频精品6学生| 国产午夜福利久久久久久| 热re99久久精品国产66热6| 我的女老师完整版在线观看| 欧美高清成人免费视频www| 少妇猛男粗大的猛烈进出视频 | 18禁动态无遮挡网站| xxx大片免费视频| 十八禁网站网址无遮挡 | 久久韩国三级中文字幕| 久久久亚洲精品成人影院| 在线亚洲精品国产二区图片欧美 | www.av在线官网国产| 99久国产av精品国产电影| 亚洲熟女精品中文字幕| 亚洲av成人精品一区久久| 深爱激情五月婷婷| 天堂中文最新版在线下载 | 午夜日本视频在线| 高清在线视频一区二区三区| 舔av片在线| 国产有黄有色有爽视频| 免费看av在线观看网站| 在线观看三级黄色| 欧美激情久久久久久爽电影| 亚洲国产精品成人久久小说| 最新中文字幕久久久久| 赤兔流量卡办理| 国模一区二区三区四区视频| 免费看a级黄色片| 免费黄频网站在线观看国产| 午夜亚洲福利在线播放| 永久免费av网站大全| 成人亚洲精品av一区二区| 国产成人福利小说| 人人妻人人澡人人爽人人夜夜| 51国产日韩欧美| www.色视频.com| 熟女av电影| 国内精品宾馆在线| 看非洲黑人一级黄片| 成人午夜精彩视频在线观看| 欧美成人一区二区免费高清观看| 久久久午夜欧美精品| 国产 一区 欧美 日韩| 午夜亚洲福利在线播放| 边亲边吃奶的免费视频| 亚洲不卡免费看| 舔av片在线| 国产午夜福利久久久久久| 女的被弄到高潮叫床怎么办| 欧美另类一区| 久久久久久久久久人人人人人人| 韩国高清视频一区二区三区| 在线观看三级黄色| 亚洲色图av天堂| 久久人人爽人人片av| 爱豆传媒免费全集在线观看| 黄色配什么色好看| 亚洲精品第二区| 18禁裸乳无遮挡动漫免费视频 | 大香蕉97超碰在线| 免费观看a级毛片全部| a级一级毛片免费在线观看| 亚洲精品影视一区二区三区av| 国产亚洲5aaaaa淫片| 成人美女网站在线观看视频| 在线观看免费高清a一片| 国产极品天堂在线| 免费少妇av软件| 毛片一级片免费看久久久久| 免费看a级黄色片| 精品一区二区三卡| 亚洲精品影视一区二区三区av| 自拍偷自拍亚洲精品老妇| 青春草视频在线免费观看| 亚洲婷婷狠狠爱综合网| 日韩欧美精品v在线| 美女被艹到高潮喷水动态| 最后的刺客免费高清国语| 日本色播在线视频| av专区在线播放| 丰满少妇做爰视频| 免费观看a级毛片全部| 赤兔流量卡办理| 久久国内精品自在自线图片| 又大又黄又爽视频免费| 免费av观看视频| 国产老妇伦熟女老妇高清| 久久久成人免费电影| 十八禁网站网址无遮挡 | 亚洲av免费高清在线观看| 精品国产三级普通话版| 人妻一区二区av| 亚洲成人久久爱视频| 美女脱内裤让男人舔精品视频|