• <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 ?

    lncreased macrophage inflammation response in pancreatic cancer patients with a diagnosis of Shi-Re

    2021-11-04 02:53:16YawenGengShulinYuLingQianKunChenYaleiZhangYeLiPengWang
    TMR Modern Herbal Medicine 2021年4期

    Yawen Geng , Shulin Yu , Ling Qian, Kun Chen , Yalei Zhang, Ye Li*, Peng Wang*

    1 Department of Integrative Oncology, Fudan University Shanghai Cancer Center, Shanghai 200032, China.

    2 Department of Oncology, Shanghai Medical College, Fudan University, Shanghai 2000332, China.

    Abstract Objective:Traditional Chinese medicine (TCM) is a comprehensive system of medical practice.ZHENG (also known as syndrome differentiation) is the essence of TCM; however, its molecular basis remains unknown.This study evaluated the molecular basis of Shi-Re ZHENG in patients with pancreatic cancer.Methods: A total of 144 patients with pathologically-confirmed locally advanced or metastatic pancreatic ductal adenocarcinoma (PDAC) were retrospectively recruited between June 2015 and February 2016.Two cohorts, including the discovery cohort (n = 60) and validation cohort (n = 84), were included in this study.QPlex multiplex array and an enzyme-linked immunosorbent assay were used to evaluate serum inflammatory cytokine levels of pancreatic cancer patients with different TCM ZHENG.Receiver operating characteristic (ROC) curve analysis was used to evaluate the importance of the systemic inflammation response index (SIRI) in the detection of Shi-Re ZHENG.Results:Shi-Re ZHENG patients exhibited a different expression pattern of cytokines, including interleukin (IL)-6, IL-8, chemokine (C-C motif) ligand 18 (CCL18), and C-reactive protein, than patients with other ZHENG diagnoses.M2-like macrophage-related inflammatory cytokines, such as IL-10, CCL17, and CCL22, were increased in Shi-Re ZHENG compared with non-Shi-Re ZHENGpatients.The SIRI score was significantly increased in Shi-Re ZHENG compared with non-Shi-Re ZHENG patients (P < 0.001).ROC analysis revealed that SIRI had a diagnostic value for Shi-Re ZHENG both in the discovery cohort (area under the curve [AUC] = 0.833, 95% confidence interval [CI] = 0.709–0.957) and validation cohort (AUC = 0.810, 95% CI = 0.716–0.905).Additionally, pancreatic cancer patients with Shi-Re ZHENG had a significantly shorter survival time than non-Shi-Re ZHENG (P = 0.002).Conclusion:Shi-Re pancreatic cancer patients are characterized by an increased macrophage inflammation response, which may contribute to a poorer prognosis when compared to those with other ZHENG diagnoses.

    Keywords:Pancreatic cancer, Shi-Re ZHENG, Inflammation

    Background

    Pancreatic ductal adenocarcinoma (PDAC) is the fourth leading cause of cancer-related deaths globally, with a five-year survival rate of only 5% [1].Despite extensive research efforts, it is still associated with low survival rates, with more than 80% of patients presenting with end-stage disease or diagnosed with metastases [2].Many studies investigating new drugs have failed to demonstrate benefits.Moreover, there is also a lack of effective treatment for advanced pancreatic cancer, and the median survival time of patients with advanced disease is around 5–6 months [3].

    Traditional Chinese medicine (TCM) is a comprehensive system of medical practice that has contributed to disease management in Chinese populations and throughout Asia for thousands of years [4].The concept of ZHENG (also known as “syndrome”) remains the essence of the TCM system.ZHENG reflects pathological changes of the microenvironment inside the human body and reveals the intrinsic character of disease more accurately, completely, and profoundly than symptoms alone [5].It is collected through four diagnostic methods, inspection, listening and smelling, questioning, and palpation, to recognize the condition of patients and develop effective, individualized treatment strategies [6].Under the conceptual frame of TCM ZHENG diagnosis, patients with the same disease might exhibit different ZHENGs and therefore should receive different treatment strategies.Conversely, patients with different diseases might exhibit the same ZHENG and therefore should receive the same treatment strategy [7].

    A ZHENG diagnosis depends on the intuition and experience of the physician trained in TCM theory, which makes it difficult to have a standard for ZHENG differentiation and further complicates the perception of the utility and validity of the method.Therefore, an understanding of the biological process underlying TCM ZHENG is important in human disease, including pancreatic cancer.In this study, we analyzed the association between different ZHENG types and the inflammatory microenvironment of patients with pancreatic cancer.Furthermore, we also attempted to better understand how ZHENG influences the clinical outcome of PDAC with changes in the microenvironment.

    Materials and methods

    Patients

    Ethical approval for this study was obtained from the Ethics Committee of the Fudan University Shanghai Cancer Center, Shanghai, China (Ethical approval no.050432-4-1212B).All patients provided written informed consent for study participation according to institutional guidelines.A total of 144 patients diagnosed with locally advanced or metastatic pancreatic cancer were retrospectively recruited from June 2015 to February 2016.The criteria of advanced pancreatic cancer were based on the 6th edition of the International Union Against Cancer, and included tumors invading the superior mesenteric artery and/or celiac axis, similar to stage III pancreatic cancer.

    Patients underwent radiological examinations such as contrast-enhanced abdominal computerized tomography scanning, magnetic resonance imaging, and/or magnetic resonance cholangiopancreatography.Patients with acute infections such as acute pancreatitis and cholangitis were excluded from this study because acute infectious disease has been shown to affect blood cell counts.A total of 144 patients were grouped into two cohorts according to their time of enrollment, with 60 recruited into the discovery cohort and the remaining 84 enrolled in the validation cohort.

    Classification of TCM ZHENG

    TCM diagnostic information was collected from a uniform questionnaire completed by the participants, including demographic factors such as age and sex, and typical symptoms for different TCM ZHENGs.Standard symptom criteria used for TCM ZHENG classification were as described previously [8].Photographs of participant tongues were also taken to enable a more accurate classification.To ensure a standard and uniform ZHENG, the classification of every patient was performed concurrently by three TCM clinical experts.

    Blood sample collection

    Fasting venous blood samples were collected in heparinized tubes prior to conducting the cancer diagnostic interventions or treatments.Samples of serum were purified by centrifuging blood at 3000 rpm for 15 min, then the serum was collected as supernatant and cryopreserved at –80°C for further analysis.

    Detection of cytokines using Q-Plex multiplex array

    Cytokine detection was performed using the Q-Plex multiplex array (Quansys Biosciences, Logan, UT, USA) on a Q-View Imager (Quansys Biosciences), in accordance with the manufacturer’s protocol, as we reported previously [9].Briefly, a series of 1:2 dilutions with known concentrations of various cytokines were performed to generate calibration curves.Fifty microliters of calibrator and samples from each well were added to the Q-Plex? Array 96-well plates and incubated at room temperature for 1 hour; the plateswere then washed and incubated with the test mixture at room temperature for 1 hour.Next, the plate was incubated with streptavidin-HRP and substrate mix at room temperature for 15 minutes.Finally, an exposure time of 270 seconds was used to image on the QView? Imager LS.The concentration of cytokines was analyzed by Q-View? software.

    Enzyme-linked immunosorbent assay (ELISA)

    The levels of interleukin (IL)-10, C-C motif chemokine ligand 17 (CCL17), and CCL22 were measured using a sandwich ELISA kit (DuoSet; R&D Systems, Minneapolis, MN) according to the manufacturer’s instructions.

    Laboratory measurements

    Routine laboratory measurements, including counts of neutrophils (N), lymphocytes (L), and monocytes (M) were carried out prior to conducting cancer diagnostic interventions or treatments.Subsequently, the systemic inflammation response index (SIRI) was calculated as follows: SIRI = N*M/L.The SIRI cutoff value for predicting Shi-Re ZHENG in the discovery cohort was calculated via receiver operating characteristic (ROC) analysis.

    Statistical analyses

    Statistical analyses were conducted using the Statistical Package for the Social Sciences (SPSS) version 15.0 (SPSS, Inc.).All data are presented as means ± standard deviation.Differences between two groups were compared using the Student’s t-test.Overall survival (OS) was determined as the time interval from initial diagnosis until death.The OS of patients in different groups was analyzed by the Kaplan–Meier curve, and survival differences were estimated using the log-rank test.P< 0.05 were regarded as statistically significant.

    Results

    Clinical–pathological features

    A total of 144 patients with pancreatic cancer were included in the present study.Of these, 96 (66.7%) were aged ≥60 years old, and 93 (64.6%) were male.Most of the tumors were located in the body-tail of the pancreas (n = 83, 57.6%), with the remainder located in the head of the pancreas (n = 61, 42.4%).

    According to the American Joint Committee on Cancer staging system, 34.7% (n = 50) of patients had stage III cancer and 65.3% (n = 94) had stage IV.CA 19-9 levels were ≥ 1,000 IU/ml in 59.0% (n = 85) of patients and < 1,000 IU/ml in 41% (n = 59) of patients.We divided all patients into four TCM ZHENGs: 47 (32.6%) patients in Shi-Re ZHENG, 39 (27.1%) in Pi-Xu ZHENG, 30 (20.8%) in Xue-Yu ZHENG, and 28 (19.4%) patients who could not be grouped into the other three ZHENG categories as Other ZHENG.Detailed clinicopathological characteristics are summarized in Table 1.

    Table 1.Clinicopathological characteristics of pancreatic cancer patients.

    Inflammatory cytokine levels

    Inflammatory cytokine levels were evaluated in 60 pancreatic cancer patients in the discovery cohort, including 15 patients of each TCM ZHENG group.Cytokines CXCL13, CCL5, and EGF showed increased expression in the Pi-Xu group and GM-CSF was increased in the Xue-Yu group (Figure 1A).The Shi-Re group showed the greatest difference in the expression of cytokines, including IL-6, IL-8, CCL18, and CRP (Figure 1A, B).These findings indicated that TCM ZHENG was associated with an altered tumor inflammatory microenvironment in pancreatic cancer patients.

    Macrophage-derived inflammatory cytokine levels

    Cytokines showing increased expression in Shi-Re ZHENG patients were mainly M2-like macrophagederived.Indeed, we observed a significant increase in the expression of M2 macrophage-derived cytokines IL-10, CCL17, and CCL22 in Shi-Re ZHENG patients compared with non-Shi-Re ZHENG patients (P< 0.001,P= 0.003, andP= 0.040, respectively; Figure 2A–C).

    We previously developed a systemic inflammation response index, SIRI, to reflect the status of the local immune response and systemic inflammation.[10]Here, we evaluated the correlation between SIRI and Shi-Re ZHENG, and found that SIRI significantly increased in Shi-Re ZHENG patients compared with non-Shi-Re ZHENG patients (P< 0.001; Figure 2D).ROC analysis showed that SIRI had a diagnostic value for Shi-Re ZHENG in advanced pancreatic cancer patients at a cutoff of 3.25 (AUC = 0.833, 95% CI = 0.709–0.957; Figure 2E).

    Figure 1.The expression of inflammatory cytokine levels in advanced pancreatic cancer patients with different ZHENGs.

    Figure 2.The macrophage inflammation response in pancreatic cancer patients with Shi-Re ZHENG and non-Shi-Re ZHENG.

    Figure 3.The clinical value of SIRI in the detection of Shi-Re ZHENG.

    SIRI clinical value

    We further evaluated the value of SIRI in the detection of Shi-Re ZHENG in the validation cohort, in which Shi-Re ZHENG accounted for 38.1%, Pi-Xu ZHENG for 28.6%, Xue-Yu ZHENG for 17.9%, and Other for 15.5% of patients (Figure 3A).SIRI was found to be significantly higher in Shi-Re ZHENG compared with other ZHENG patients (P< 0.001; Figure 3B).Comparing the sensitivity and specificity of ROC analysis in the discovery cohort, SIRI was shown to predict Shi-Re ZHENG at a cutoff of 3.25.Using ROC analysis at a cutoff of 3.25 revealed that SIRI also had a diagnostic value for Shi-Re ZHENG in the validation cohort (AUC = 0.810, 95% CI = 0.716–0.905; Figure 3C).

    Shi-Re ZHENG and prognosis

    We next examined the correlation between SIRI and OS in the validation cohort of 84 patients.The median OS for patients in this cohort was 3.5 months for those with SIRI scores ≥ 3.5 and 7.6 months for those with SIRI scores <3.5 (P< 0.001; Figure 4A), indicating a significantly shorter survival time in the former group.Kaplan–Meier analyses further demonstrated the association between Shi-Re ZHENG and OS, with the median OS being significantly shorter (4.5 months) in Shi-Re ZHENG pancreatic cancer patients compared with those in other ZHENG groups (8.8 months for Pi-Xu ZHENG, 6.7 months for Xue-Yu ZHENG, and 7.2 months for Other) (P= 0.002; Figure 4B).

    Figure 4.Shi-Re ZHENG was associated with poor prognosis.

    Discussion

    ZHENG, also known as the TCM pattern or TCM syndrome, is an essential and integral component of the TCM theory.A recent study demonstrated that Shi-Re ZHENG patients with colorectal cancer expressed higher levels of inflammatory cytokines (IL-1a, cyclooxygenase-2, and CCL-2) and proliferationassociated transcription factors (Ki-67 and proliferating cell nuclear antigen) than Pi-Xu ZHENG patients and had a worse survival [11].Moreover, Yinet alestablished a pancreatic cancer xenograft mouse model and demonstrated altered tumor-associated macrophage infiltration and cancer-associated myofibroblast proliferative activities in Shi-Re ZHENG mice.[12]The application of ZHENG in cancer patients could not only enhance the efficacy of treatment but also affect the clinical outcome.[13]In our study, we found that Shi-Re is the most common ZHENG in pancreatic cancer patients, and that these patients have a poor prognosis compared with those with other ZHENG types.

    Cancer-related inflammation has been proposed as the seventh hallmark of cancer.[14]Several pieces of evidence have shown that the tumor microenvironment (TME) engages in cancer initiation and the promotion of tumor growth.The PDAC microenvironment is characterized by a dense desmoplastic stroma [15] which accounts for up to 50%–80% of the tumor volume [16].Tumor-associated macrophages (TAMs) are the most dominant stromal cell population in the TME of many tumors, including PDAC.TAMs were first described in the early 1980s, and their importance in promoting tumor initiation and development has been increasingly recognized, with their infiltration into tumors recently being correlated with poor prognosis [17-19].TAM-expressed CCL18 induces cancer cell epithelial–mesenchymal transition (EMT) via a feedback loop in breast cancer [20],while TAMs also promote EMT in pancreatic cancer by activating the Toll-like receptor 4/IL-10 signaling pathway [21, 22].Several TAM-secreted cytokines (including IL-4, IL-10, hepatocyte growth factor, vascular endothelial growth factor, and transforming growth factor-β) and chemokines (such as CCL17, CCL18, CCL22, CCL2, and CCL5) function in immunosuppression, proinflammation, angiogenesis, and metastasis [23, 24].In our research, we demonstrated that Shi-Re ZHENG patients exhibited higher serum levels of TAM-derived cytokines including IL-10, CCL17, CCL18, and CCL22.We also observed differences in cytokine expression among pancreatic cancer patients with different TCM ZHENG.In particular, those in the Shi-Re ZHENG group showed differences in the expression of cytokines associated with an M2-like macrophage inflammation response.

    We previously developed a novel systemic inflammation response index, SIRI, based on the counts of monocytes, lymphocytes, and peripheral neutrophils, which can be applied to predict the survival of pancreatic cancer patients treated with chemotherapy, and improve the efficacy of individualized treatments [10].Its prediction ability is stronger than that of other inflammatory indexes, including the neutrophil: lymphocyte ratio and platelet: lymphocyte ratio, and is related to higher serum levels of inflammatory cytokines/chemokines IL-10, CCL17, CCL18, and CCL22.High SIRI scores are indicative of poor outcomes, and their value in assessing the prognosis of patients was also demonstrated in other carcinomas such as nasopharyngeal carcinoma and esophageal squamous cell carcinoma [25, 26].SIRI could be used to predict treatment responses to chemotherapy as an “immunologic signature” in different types of cancers, which would help clinicians identify those patients who would most benefit most therapy [27].Here, we demonstrated a correlation between SIRI and Shi-Re ZHENG, with Shi-Re ZHENG patients having higher SIRI values associated with increased serum levels of inflammatory cytokines/chemokines, and significantly shorter outcomes than other patients with advanced pancreatic cancer.

    A main strength of this study lies in our attempt to elucidate TCM ZHENG within the perspective of an inflammatory microenvironment.We demonstrated that different types of cytokines are expressed in different TCM ZHENGs, suggesting a molecular basis for ZHENG.TAMs have been confirmed to suppress immune responses and believed to contribute to tumor invasion and progression [28].TAM-related inflammatory cytokines, like IL-6, IL-10, IL-17 and IL-22 are also involved in process [29, 30].An imbalanced adjustment in TAM-related inflammatory cytokines has been regarded as an important pro-inflammatory regulator in the tumor microenvironment.We previously showed that the infiltration of TAMs and the activity of cancer-associated fibroblasts (CAFs) were altered under different ZHENGs condition.Accordingly, the expression levels of TAM-derived cytokines and CAF-derived chemokines were altered, such as TAM-related CCL5/CCR5 and CAF-related SDF-1/CXCR4 [31].Based on the roles of TAMs and related inflammatory factors in cancer, we proposed that the TAM infiltration and TAM-related inflammatory cytokines might be the key to influencing ZHENGs.

    We further explained the relationship between Shi-Re ZHENG and the inflammatory microenvironment using the previously established inflammatory index, SIRI, to show that SIRI could be used for the diagnosis of Shi-Re ZHENG.Most importantly, ZHENG was found not only to exist as a concept, but also to have potential clinical value for the outcome of pancreatic cancer.A study limitation is that the research is a preliminary exploratory study.However, the future application of multi-omics technology will help explain the biological basis of ZHENG more systematically and comprehensively.

    Conclusion

    Shi-Re pancreatic cancer patients are characterized by an increased macrophage inflammation response, which may contribute to a poorer prognosis when compared to those with other ZHENG diagnoses.

    天天影视国产精品| 日韩免费av在线播放| 黑人猛操日本美女一级片| 成年女人毛片免费观看观看9 | 亚洲成国产人片在线观看| 两人在一起打扑克的视频| 欧美成狂野欧美在线观看| 欧美亚洲 丝袜 人妻 在线| 18禁裸乳无遮挡免费网站照片 | 怎么达到女性高潮| 国精品久久久久久国模美| 中文字幕av电影在线播放| 中文字幕最新亚洲高清| 人人澡人人妻人| 国产成人精品久久二区二区免费| 交换朋友夫妻互换小说| 久久精品人人爽人人爽视色| 国产亚洲精品久久久久5区| 亚洲欧美一区二区三区黑人| 亚洲av电影在线进入| 91精品三级在线观看| 国产一区二区三区综合在线观看| 国产真人三级小视频在线观看| 一a级毛片在线观看| 三上悠亚av全集在线观看| 久久人人97超碰香蕉20202| 国产野战对白在线观看| av中文乱码字幕在线| 嫩草影视91久久| 天天躁夜夜躁狠狠躁躁| 纯流量卡能插随身wifi吗| 丝袜在线中文字幕| 午夜日韩欧美国产| 少妇猛男粗大的猛烈进出视频| www.熟女人妻精品国产| 国产激情久久老熟女| 亚洲avbb在线观看| 亚洲精品国产一区二区精华液| 国产精品偷伦视频观看了| 国产精品一区二区在线观看99| 精品人妻在线不人妻| 午夜免费成人在线视频| 国产精品影院久久| 女人高潮潮喷娇喘18禁视频| av线在线观看网站| 丝瓜视频免费看黄片| a级毛片黄视频| 一本综合久久免费| 91精品国产国语对白视频| 国产aⅴ精品一区二区三区波| av天堂久久9| 国产人伦9x9x在线观看| 黑人猛操日本美女一级片| 久久久久久亚洲精品国产蜜桃av| 亚洲第一欧美日韩一区二区三区| 国产精品永久免费网站| 精品久久久精品久久久| 亚洲专区中文字幕在线| 99精品欧美一区二区三区四区| 香蕉丝袜av| 国产av又大| 国产男女内射视频| 午夜91福利影院| 老司机亚洲免费影院| 亚洲专区中文字幕在线| 在线天堂中文资源库| 啪啪无遮挡十八禁网站| 首页视频小说图片口味搜索| 日日爽夜夜爽网站| 老司机在亚洲福利影院| av天堂在线播放| 亚洲,欧美精品.| 69av精品久久久久久| 在线观看免费午夜福利视频| 高清在线国产一区| 亚洲专区国产一区二区| 搡老岳熟女国产| 国产精品一区二区免费欧美| 9热在线视频观看99| 久久久久久久国产电影| 亚洲精品美女久久av网站| 99国产精品一区二区蜜桃av | 免费日韩欧美在线观看| 精品亚洲成a人片在线观看| 精品亚洲成a人片在线观看| 成人国语在线视频| 一级片免费观看大全| 岛国毛片在线播放| 国产不卡av网站在线观看| 午夜福利,免费看| 91麻豆精品激情在线观看国产 | 国产aⅴ精品一区二区三区波| 又黄又粗又硬又大视频| 亚洲成人免费av在线播放| 日韩精品免费视频一区二区三区| av天堂在线播放| 一区二区三区激情视频| 十分钟在线观看高清视频www| 777米奇影视久久| 国产精品免费大片| 欧美国产精品一级二级三级| 国产精品一区二区免费欧美| 91九色精品人成在线观看| 天天躁日日躁夜夜躁夜夜| 天天躁日日躁夜夜躁夜夜| 久久精品亚洲熟妇少妇任你| 麻豆成人av在线观看| 黄色 视频免费看| 国产一区在线观看成人免费| 啦啦啦 在线观看视频| 丝袜人妻中文字幕| 色综合欧美亚洲国产小说| 亚洲色图综合在线观看| 久热爱精品视频在线9| 欧美日韩亚洲高清精品| 丝瓜视频免费看黄片| 一级a爱片免费观看的视频| 久久中文看片网| 女性生殖器流出的白浆| 久久99一区二区三区| 午夜福利在线观看吧| 久久午夜亚洲精品久久| 国产97色在线日韩免费| svipshipincom国产片| 国产男女内射视频| 久久影院123| 成人精品一区二区免费| 女性被躁到高潮视频| 身体一侧抽搐| 免费黄频网站在线观看国产| 波多野结衣av一区二区av| 美女视频免费永久观看网站| 久热爱精品视频在线9| 亚洲欧美日韩另类电影网站| 在线观看66精品国产| 国产高清视频在线播放一区| 国产午夜精品久久久久久| a在线观看视频网站| 国产片内射在线| 亚洲午夜精品一区,二区,三区| 精品亚洲成a人片在线观看| 国产一区二区激情短视频| 久9热在线精品视频| 欧美久久黑人一区二区| 不卡一级毛片| 亚洲精品美女久久av网站| 最新的欧美精品一区二区| 久久久久精品国产欧美久久久| 欧美不卡视频在线免费观看 | 成年人免费黄色播放视频| 欧美成狂野欧美在线观看| 亚洲欧洲精品一区二区精品久久久| 久久精品亚洲熟妇少妇任你| 国产99白浆流出| 亚洲男人天堂网一区| 一级,二级,三级黄色视频| 熟女少妇亚洲综合色aaa.| 欧美性长视频在线观看| 国产成人系列免费观看| 人成视频在线观看免费观看| 国产高清国产精品国产三级| 午夜激情av网站| 精品人妻在线不人妻| 老鸭窝网址在线观看| 人妻久久中文字幕网| 免费在线观看视频国产中文字幕亚洲| 天堂俺去俺来也www色官网| 另类亚洲欧美激情| 欧美成狂野欧美在线观看| 男女高潮啪啪啪动态图| 国产视频一区二区在线看| 国产精品国产av在线观看| 老鸭窝网址在线观看| 伦理电影免费视频| 在线观看免费视频日本深夜| 国产成人系列免费观看| 亚洲七黄色美女视频| 精品欧美一区二区三区在线| 欧美 亚洲 国产 日韩一| 日韩成人在线观看一区二区三区| 黄频高清免费视频| 成熟少妇高潮喷水视频| 一边摸一边做爽爽视频免费| 亚洲av美国av| 久久精品熟女亚洲av麻豆精品| 大香蕉久久网| 久久香蕉激情| av不卡在线播放| 亚洲美女黄片视频| 国产伦人伦偷精品视频| www.熟女人妻精品国产| 女人被躁到高潮嗷嗷叫费观| 人人妻人人爽人人添夜夜欢视频| 欧美精品一区二区免费开放| 免费在线观看日本一区| 99久久人妻综合| 亚洲久久久国产精品| 最新的欧美精品一区二区| 人人澡人人妻人| 美女视频免费永久观看网站| 国产三级黄色录像| 日本vs欧美在线观看视频| 午夜福利影视在线免费观看| 啦啦啦免费观看视频1| 啪啪无遮挡十八禁网站| 免费在线观看日本一区| 在线视频色国产色| 免费看十八禁软件| 看免费av毛片| 自线自在国产av| 露出奶头的视频| 男人操女人黄网站| 久久青草综合色| 91字幕亚洲| 美女高潮到喷水免费观看| 丝袜人妻中文字幕| 亚洲国产中文字幕在线视频| 好看av亚洲va欧美ⅴa在| av中文乱码字幕在线| 欧美精品人与动牲交sv欧美| 91精品国产国语对白视频| 精品视频人人做人人爽| av片东京热男人的天堂| 亚洲va日本ⅴa欧美va伊人久久| 伊人久久大香线蕉亚洲五| 大型av网站在线播放| 国产精品一区二区免费欧美| 巨乳人妻的诱惑在线观看| 国产精品久久视频播放| av一本久久久久| 建设人人有责人人尽责人人享有的| 精品一区二区三卡| 日韩欧美免费精品| tocl精华| 大陆偷拍与自拍| a级毛片在线看网站| 中文字幕最新亚洲高清| 97人妻天天添夜夜摸| 精品一区二区三卡| 免费看十八禁软件| 亚洲色图综合在线观看| 天天躁狠狠躁夜夜躁狠狠躁| 交换朋友夫妻互换小说| 最新美女视频免费是黄的| 亚洲三区欧美一区| 女性生殖器流出的白浆| 日本vs欧美在线观看视频| 手机成人av网站| 在线观看免费视频网站a站| 超色免费av| 最近最新中文字幕大全电影3 | 高潮久久久久久久久久久不卡| 热re99久久国产66热| 国产精品影院久久| 国产欧美亚洲国产| x7x7x7水蜜桃| 欧美日韩瑟瑟在线播放| av视频免费观看在线观看| 欧美日韩亚洲综合一区二区三区_| 少妇的丰满在线观看| 成年版毛片免费区| 午夜福利一区二区在线看| 国产黄色免费在线视频| 最新美女视频免费是黄的| 久久婷婷成人综合色麻豆| 欧美日本中文国产一区发布| 青草久久国产| 亚洲中文日韩欧美视频| 亚洲伊人色综图| 1024香蕉在线观看| 黄频高清免费视频| 日韩一卡2卡3卡4卡2021年| 美女福利国产在线| 韩国精品一区二区三区| www.999成人在线观看| 国产精品偷伦视频观看了| 免费观看精品视频网站| 国产亚洲精品久久久久久毛片 | x7x7x7水蜜桃| 欧美国产精品va在线观看不卡| av欧美777| 欧美亚洲 丝袜 人妻 在线| 高潮久久久久久久久久久不卡| 日韩制服丝袜自拍偷拍| 国产精品av久久久久免费| 少妇的丰满在线观看| 又黄又爽又免费观看的视频| 老司机影院毛片| 精品第一国产精品| 另类亚洲欧美激情| 国内久久婷婷六月综合欲色啪| 亚洲少妇的诱惑av| 国产成人av教育| 日本黄色视频三级网站网址 | 久久久久久免费高清国产稀缺| 亚洲欧美一区二区三区黑人| 亚洲专区字幕在线| 日韩欧美三级三区| 国产精品美女特级片免费视频播放器 | 成在线人永久免费视频| 少妇被粗大的猛进出69影院| 国产有黄有色有爽视频| 国产精品亚洲av一区麻豆| 岛国在线观看网站| 欧美黑人欧美精品刺激| 99热网站在线观看| 香蕉久久夜色| 精品人妻1区二区| 久久精品人人爽人人爽视色| 亚洲色图 男人天堂 中文字幕| 高潮久久久久久久久久久不卡| 久久久久久久国产电影| 免费观看人在逋| 欧美国产精品一级二级三级| 亚洲熟女精品中文字幕| 精品少妇一区二区三区视频日本电影| 夜夜爽天天搞| www日本在线高清视频| 90打野战视频偷拍视频| 男男h啪啪无遮挡| 亚洲久久久国产精品| 欧美精品av麻豆av| 一区二区日韩欧美中文字幕| 人妻久久中文字幕网| 新久久久久国产一级毛片| 亚洲人成电影免费在线| 午夜视频精品福利| 日本一区二区免费在线视频| 丰满饥渴人妻一区二区三| 久久精品国产亚洲av高清一级| 黄色丝袜av网址大全| 精品高清国产在线一区| 熟女少妇亚洲综合色aaa.| 99国产精品一区二区三区| 欧美激情久久久久久爽电影 | 人人妻人人澡人人看| 亚洲一区二区三区不卡视频| 搡老岳熟女国产| 啪啪无遮挡十八禁网站| 淫妇啪啪啪对白视频| 涩涩av久久男人的天堂| 1024视频免费在线观看| 高清在线国产一区| 老司机靠b影院| а√天堂www在线а√下载 | 国产精品二区激情视频| 欧美人与性动交α欧美精品济南到| 亚洲国产精品一区二区三区在线| 男女之事视频高清在线观看| 国产单亲对白刺激| 午夜激情av网站| 99在线人妻在线中文字幕 | 成人精品一区二区免费| 99久久人妻综合| 成人免费观看视频高清| 桃红色精品国产亚洲av| 女人爽到高潮嗷嗷叫在线视频| 国产又色又爽无遮挡免费看| 成年人黄色毛片网站| 亚洲国产精品合色在线| 成年人免费黄色播放视频| 欧美日韩亚洲国产一区二区在线观看 | 国产精品国产高清国产av | 丰满人妻熟妇乱又伦精品不卡| 男人舔女人的私密视频| 99国产精品一区二区蜜桃av | 久久久国产精品麻豆| 丰满人妻熟妇乱又伦精品不卡| av天堂在线播放| 最近最新免费中文字幕在线| 50天的宝宝边吃奶边哭怎么回事| 国产深夜福利视频在线观看| 1024视频免费在线观看| 亚洲av片天天在线观看| 午夜免费观看网址| 夜夜夜夜夜久久久久| 国产亚洲av高清不卡| 中文字幕人妻丝袜制服| 亚洲自偷自拍图片 自拍| 久久国产精品大桥未久av| 亚洲午夜理论影院| 欧美老熟妇乱子伦牲交| 精品久久蜜臀av无| 999精品在线视频| 黄色视频不卡| 亚洲国产中文字幕在线视频| 日韩免费av在线播放| 亚洲国产欧美一区二区综合| 国产成人免费无遮挡视频| 国产淫语在线视频| 国内久久婷婷六月综合欲色啪| 一区二区日韩欧美中文字幕| 久久午夜综合久久蜜桃| 99re6热这里在线精品视频| 高潮久久久久久久久久久不卡| 国产精品二区激情视频| 亚洲色图综合在线观看| 午夜福利在线免费观看网站| 香蕉久久夜色| 交换朋友夫妻互换小说| 国产亚洲精品久久久久久毛片 | 黑人巨大精品欧美一区二区蜜桃| 免费不卡黄色视频| 丁香欧美五月| www.精华液| 午夜免费观看网址| 亚洲欧美日韩高清在线视频| 成人免费观看视频高清| 搡老熟女国产l中国老女人| 叶爱在线成人免费视频播放| 热99国产精品久久久久久7| 亚洲av日韩在线播放| 美女国产高潮福利片在线看| 国产成人影院久久av| 国产视频一区二区在线看| 超色免费av| 超碰97精品在线观看| 午夜精品久久久久久毛片777| 大型黄色视频在线免费观看| 十分钟在线观看高清视频www| tube8黄色片| 熟女少妇亚洲综合色aaa.| 精品人妻在线不人妻| 国产精品一区二区在线不卡| 亚洲久久久国产精品| 久久 成人 亚洲| 一个人免费在线观看的高清视频| av电影中文网址| 亚洲七黄色美女视频| 成年版毛片免费区| 成年人黄色毛片网站| 三上悠亚av全集在线观看| 国产精品秋霞免费鲁丝片| 在线视频色国产色| 国产精品 欧美亚洲| 欧美激情 高清一区二区三区| 人人妻人人添人人爽欧美一区卜| 亚洲人成77777在线视频| 国产在视频线精品| 亚洲国产欧美一区二区综合| 精品国产超薄肉色丝袜足j| 一级作爱视频免费观看| 色94色欧美一区二区| 成熟少妇高潮喷水视频| 亚洲国产欧美日韩在线播放| 亚洲av日韩在线播放| 美女福利国产在线| 国产不卡av网站在线观看| 女人被狂操c到高潮| 国产欧美日韩一区二区三区在线| 亚洲五月天丁香| 欧美乱码精品一区二区三区| 亚洲av成人av| 在线播放国产精品三级| 91麻豆精品激情在线观看国产 | 精品一区二区三区视频在线观看免费 | 51午夜福利影视在线观看| 欧美国产精品va在线观看不卡| 男女下面插进去视频免费观看| 国产亚洲精品久久久久久毛片 | 国产日韩欧美亚洲二区| 精品欧美一区二区三区在线| 美女午夜性视频免费| 91大片在线观看| 人人妻人人澡人人看| 午夜精品国产一区二区电影| 精品久久蜜臀av无| 一区福利在线观看| 精品国内亚洲2022精品成人 | 自拍欧美九色日韩亚洲蝌蚪91| 欧美精品高潮呻吟av久久| 国产精品久久电影中文字幕 | 久久草成人影院| 国产xxxxx性猛交| 国产片内射在线| 国产欧美亚洲国产| 亚洲专区字幕在线| 久久精品91无色码中文字幕| 欧美国产精品va在线观看不卡| 日韩大码丰满熟妇| 丝瓜视频免费看黄片| 成年女人毛片免费观看观看9 | 99久久综合精品五月天人人| 国产免费av片在线观看野外av| 高潮久久久久久久久久久不卡| 久久香蕉国产精品| 国产免费男女视频| 在线观看免费日韩欧美大片| 黑人巨大精品欧美一区二区蜜桃| 大片电影免费在线观看免费| 久久中文看片网| 真人做人爱边吃奶动态| 51午夜福利影视在线观看| 国产精品一区二区精品视频观看| 欧美色视频一区免费| 精品亚洲成国产av| 欧美大码av| 国产一区二区三区在线臀色熟女 | 人妻 亚洲 视频| 黄色怎么调成土黄色| 一个人免费在线观看的高清视频| 欧美精品啪啪一区二区三区| 十八禁人妻一区二区| 乱人伦中国视频| 熟女少妇亚洲综合色aaa.| 男女床上黄色一级片免费看| 亚洲av成人一区二区三| 天天操日日干夜夜撸| 精品国产超薄肉色丝袜足j| 女警被强在线播放| 免费黄频网站在线观看国产| 久久热在线av| 国产三级黄色录像| 激情在线观看视频在线高清 | 婷婷精品国产亚洲av在线 | 久久香蕉国产精品| www日本在线高清视频| 高清毛片免费观看视频网站 | 美女午夜性视频免费| 亚洲情色 制服丝袜| 精品国产一区二区三区久久久樱花| 一边摸一边抽搐一进一出视频| 久久九九热精品免费| 免费看a级黄色片| 色尼玛亚洲综合影院| 欧美激情极品国产一区二区三区| 欧美精品啪啪一区二区三区| 两性夫妻黄色片| 亚洲精品在线观看二区| 欧美国产精品一级二级三级| videosex国产| 国产乱人伦免费视频| 两性夫妻黄色片| 亚洲av成人不卡在线观看播放网| 高清av免费在线| 亚洲精品国产精品久久久不卡| 免费日韩欧美在线观看| 波多野结衣一区麻豆| 在线视频色国产色| 人妻丰满熟妇av一区二区三区 | 啦啦啦视频在线资源免费观看| 在线看a的网站| 亚洲三区欧美一区| 两人在一起打扑克的视频| 国产欧美日韩一区二区三| 在线观看免费高清a一片| 欧美在线一区亚洲| 国产精品综合久久久久久久免费 | 国产精品久久久久久人妻精品电影| 精品国产超薄肉色丝袜足j| 国产97色在线日韩免费| 欧美激情 高清一区二区三区| 男男h啪啪无遮挡| 老鸭窝网址在线观看| 嫩草影视91久久| 欧美中文综合在线视频| 露出奶头的视频| 精品少妇一区二区三区视频日本电影| 日本wwww免费看| 欧美午夜高清在线| 精品国产国语对白av| 亚洲五月婷婷丁香| tube8黄色片| 免费观看精品视频网站| av网站免费在线观看视频| 12—13女人毛片做爰片一| 国产麻豆69| av在线播放免费不卡| 中文字幕av电影在线播放| 国产精品久久视频播放| 好男人电影高清在线观看| 国产精品欧美亚洲77777| 色94色欧美一区二区| 国产亚洲精品久久久久久毛片 | 一区福利在线观看| 亚洲精品av麻豆狂野| 国产成人精品久久二区二区91| 女人被躁到高潮嗷嗷叫费观| 欧洲精品卡2卡3卡4卡5卡区| 久久国产精品大桥未久av| 午夜两性在线视频| 久久久久国产一级毛片高清牌| 在线免费观看的www视频| 免费观看精品视频网站| 高清黄色对白视频在线免费看| 伊人久久大香线蕉亚洲五| 制服人妻中文乱码| 国产一区二区三区在线臀色熟女 | 精品一品国产午夜福利视频| 一级毛片高清免费大全| 18禁裸乳无遮挡免费网站照片 | 日韩大码丰满熟妇| 午夜福利视频在线观看免费| 亚洲在线自拍视频| 男女下面插进去视频免费观看| 亚洲av成人不卡在线观看播放网| 中文字幕最新亚洲高清| 久久精品国产亚洲av高清一级| 久久人人97超碰香蕉20202| 国产精品一区二区在线不卡| 亚洲av第一区精品v没综合| 一进一出抽搐gif免费好疼 | 麻豆成人av在线观看| av片东京热男人的天堂| 欧美日本中文国产一区发布| 变态另类成人亚洲欧美熟女 | 精品一区二区三区四区五区乱码| 91麻豆精品激情在线观看国产 | av在线播放免费不卡| 中文字幕高清在线视频| 国产精华一区二区三区| 亚洲国产毛片av蜜桃av| 美女福利国产在线| 久久性视频一级片| 欧美黑人精品巨大|