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1.
背景:胶原特殊的分子结构和生物活性有利于多种细胞黏附、增殖和分化,并可降解为新生组织提供足够空间。目的:制备一种复合负载碱性成纤维细胞生长因子的壳聚糖-肝素纳米粒子双层胶原基复合材料,并评价其生物安全性。方法:制备交联风干胶原膜和交联冻干胶原膜。将壳聚糖-肝素纳米粒滴于交联冻干胶原膜上,再将湿态交联风干胶原膜置于复合纳米粒子的交联冻干胶原膜上风干,即碱性成纤维细胞生长因子/双层胶原基复合材料。采用急性全身毒性试验、溶血试验、热原试验和细胞毒性试验评价其生物安全性。结果与结论:碱性成纤维细胞生长因子/双层胶原基复合材料为双层结构,一侧表面致密,另一侧疏松多孔。在其中间负载碱性成纤维细胞生长因子的壳聚糖-肝素纳米粒子呈不规则球形分布于胶原膜内侧面;急性全身毒性试验、热原试验、溶血试验均为阴性,细胞毒性为0级。说明碱性成纤维细胞生长因子/双层胶原基复合材料具有良好的生物安全性,对机体无毒,符合ISO10993-1评价标准。  相似文献   

2.
胶原-壳聚糖复合材料的制备及生物安全性检测   总被引:4,自引:2,他引:4  
目的:制备胶原-壳聚糖复合细胞载体,并对该载体进行系统的生物学性能检测。方法:实验于2003-07/2006-06在天津市医药科学研究所完成。将胶原溶液和壳聚糖溶液按照一定比例(9∶1)混匀,交联后真空冷冻干燥,制成胶原-壳聚糖复合载体,对该载体进行复合材料性状及理化性能检测。并采用急性全身毒性试验、皮内刺激试验、皮肤致敏试验、溶血试验、细胞毒性试验和致突变试验进行生物学性能检测。结果:①胶原-壳聚糖复合材料具有三维立体多孔结构,适合细胞的三维生长,能为新生组织提供良好的支架。②急性全身毒性试验、皮内刺激试验均阴性,致敏率仅为6.25%,细胞毒性为0级,无诱变能力。结论:从仿生学角度出发,制备出可生物降解的胶原-壳聚糖复合材料,经过系统的生物学评价发现,胶原-壳聚糖复合材料具有良好的生物安全性,对机体无毒,不致突变,对细胞有较强的亲和作用,利于细胞生长和分化。  相似文献   

3.
背景:通过各种微球负载骨生长因子使骨形态发生蛋白达到缓释效果逐渐成为研究热点,但关于载药壳聚糖纳米微球的生物相容性特别是细胞毒性的报道较少。目的:对重组人骨形态发生蛋白2壳聚糖纳米微球进行细胞毒性检测,评估应用壳聚糖纳米微球作为重组人骨形态发生蛋白2缓释载体的生物安全性。方法:通过离子交联法制备空白壳聚糖纳米微球,应用透视电镜观察微球的形态,激光粒径分析其粒径分布;通过重组人骨形态发生蛋白2壳聚糖纳米微球体外细胞毒性试验评估微球的生物安全性。结果与结论:离子交联法制备的壳聚糖微球,球形规整,分散均匀,微球平均粒径为230nm,分布较集中。载药及空白微球的反应分级为0或1级,均为合格。提示,离子交联法制备可成功制备出负载重组人骨形态发生蛋2的纳米微球,且微球细胞毒性检测合格,为进一步的骨组织工程研究提供理论实验基础。  相似文献   

4.
王玮  尹庆水  张余 《中国临床康复》2011,(25):4611-4614
背景:通过各种微球负载骨生长因子使骨形态发生蛋白达到缓释效果逐渐成为研究热点,但关于载药壳聚糖纳米微球的生物相容性特别是细胞毒性的报道较少。目的:对重组人骨形态发生蛋白2壳聚糖纳米微球进行细胞毒性检测,评估应用壳聚糖纳米微球作为重组人骨形态发生蛋白2缓释载体的生物安全性。方法:通过离子交联法制备空白壳聚糖纳米微球,应用透视电镜观察微球的形态,激光粒径分析其粒径分布;通过重组人骨形态发生蛋白2壳聚糖纳米微球体外细胞毒性试验评估微球的生物安全性。结果与结论:离子交联法制备的壳聚糖微球,球形规整,分散均匀,微球平均粒径为230nm,分布较集中。载药及空白微球的反应分级为0或1级,均为合格。提示,离子交联法制备可成功制备出负载重组人骨形态发生蛋2的纳米微球,且微球细胞毒性检测合格,为进一步的骨组织工程研究提供理论实验基础。  相似文献   

5.
背景:目前人工皮肤替代品的种类较多,各有优缺点,仍然没有一种理想的产品应用于临床。目的:探讨构建一种可以缓慢释放碱性成纤维细胞生长因子的新型人工活性真皮的可行性。方法:组织块法培养幼儿包皮成纤维细胞;采用酶-去垢剂法制备人脱细胞羊膜;双相法制备碱性成纤维细胞生长因子-明胶-壳聚糖缓释微球;缓释微球黏附于脱细胞羊膜上;三四代成纤维细胞培养于负载缓释微球的脱细胞羊膜上。结果与结论:制备的脱细胞羊膜为白色半透明状薄膜有较高的孔隙率,空隙不规则,孔径大小为10-100nm,无细胞毒性;碱性成纤维细胞生长因子-明胶-壳聚糖缓释微球分散较均匀,呈球形,粒径均匀,球体表面比较光滑,载药率为20ng/g,包封率为80.5%,体外药物缓释曲线显示药物控释效果良好;成纤维细胞在支架表面爬行生长良好,层粘连蛋白表达较对照组高。表明将成纤维细胞种植于负载碱性成纤维细胞生长因子-明胶-壳聚糖缓释微球的脱细胞羊膜上,缓释微球能较好地黏附于脱细胞羊膜表面。  相似文献   

6.
背景:目前胶原作为牙周组织工程支架材料仍具有机械强度差、降解速度快等缺点,将其与壳聚糖复合可改善上述问题。 目的:评估新型壳聚糖-胶原支架材料的体外生物相容性。 方法:通过 MTT 法评估100%,75%,50%,25%壳聚糖-胶原支架材料浸提液对人牙周膜细胞的毒性。选择第4-6代生长状态良好的人牙周膜细胞与壳聚糖-胶原支架共培养,观察细胞在支架上的生长情况,并检测与壳聚糖-胶原支架复合培养前后人牙周膜细胞碱性磷酸酶活性的变化。 结果与结论:新型壳聚糖-胶原支架具有双层结构,一侧表面致密,一侧表面疏松多孔。MTT 法检测不同浓度材料浸提液毒性评级为0或1级。扫描电子显微镜及组织学观察可见细胞在壳聚糖-胶原支架上增殖良好,且致密层可起屏障膜作用,阻挡细胞进入支架内部;复合培养24 h后,人牙周膜细胞的碱性磷酸酶活性与复合培养前无明显差异(P 〉0.05),复合培养48,72 h后人牙周膜细胞的碱性磷酸酶活性高于复合培养前(P 〈0.05)。以上结果提示新型壳聚糖-胶原支架具有良好的生物相容性及屏障功能,可进一步应用于牙周组织工程的研究。  相似文献   

7.
蒋斌  杨锋  刘志坤 《中国临床康复》2012,(34):6343-6348
背景:纳米羟基磷灰石/壳聚糖复合物具有良好的力学性能和生物相容性,可用于骨损伤的修复,是一种有应用前景的骨替代品。碱性成纤维细胞生长因子可促进组织修复,目的:观察碱性成纤维细胞生长因子复合纳米羟基磷灰石/壳聚糖复合物修复兔骨缺损的效果。方法:构建桡骨缺损兔模型,按植入材料的不同共分为3组,实验组植入纳米羟基磷灰石/壳聚糖复合物+碱性成纤维细胞生长因子,对照植入组纳米羟基磷灰石/壳聚糖复合物,空白组无任何材料植入。结果与结论:干预12周时,X射线片检查显示,实验组骨植入区新骨发生骨性融合,髓腔再通骨缺损已基本消失;苏木精-伊红染色结果显示,实验组出现成熟的板层骨、成熟的哈弗氏系统以及破骨细胞增生引起的骨质吸收区;以上结果均为实验组的修复效果优于对照组。证实,碱性成纤维细胞生长因子复合纳米羟基磷灰石/壳聚糖复合物可促进骨缺损修复,效果优于单独应用纳米羟基磷灰石/壳聚糖复合物。  相似文献   

8.
背景:纳米羟基磷灰石/壳聚糖复合物具有良好的力学性能和生物相容性,可用于骨损伤的修复,是一种有应用前景的骨替代品。碱性成纤维细胞生长因子可促进组织修复,目的:观察碱性成纤维细胞生长因子复合纳米羟基磷灰石/壳聚糖复合物修复兔骨缺损的效果。方法:构建桡骨缺损兔模型,按植入材料的不同共分为3组,实验组植入纳米羟基磷灰石/壳聚糖复合物+碱性成纤维细胞生长因子,对照植入组纳米羟基磷灰石/壳聚糖复合物,空白组无任何材料植入。结果与结论:干预12周时,X射线片检查显示,实验组骨植入区新骨发生骨性融合,髓腔再通骨缺损已基本消失;苏木精-伊红染色结果显示,实验组出现成熟的板层骨、成熟的哈弗氏系统以及破骨细胞增生引起的骨质吸收区;以上结果均为实验组的修复效果优于对照组。证实,碱性成纤维细胞生长因子复合纳米羟基磷灰石/壳聚糖复合物可促进骨缺损修复,效果优于单独应用纳米羟基磷灰石/壳聚糖复合物。  相似文献   

9.
背景:胰岛素样生长因子1具有促进成纤维细胞有丝分裂的作用,同时具有促进牙周细胞生长、分化及合成细胞外基质的作用。 目的:观察负载胰岛素样生长因子1的壳聚糖胶原支架对于人牙周膜细胞增殖的作用。 方法:将人牙周膜细胞分别接种于负载胰岛素样生长因子1的壳聚糖胶原支架与普通胶原支架上,于接种的1 h、24 h及1周检测重组人转化生长因子β1的释放,于第1,7,28天检测两组细胞的黏附和增殖情况。结果与结论:负载胰岛素样生长因子1的壳聚糖胶原支架组第1,24小时和第1周的重组人转化生长因子β1释放率明显低于普通胶原支架组(P 0.05),负载胰岛素样生长因子1的壳聚糖胶原支架组接种第7,28天的细胞黏附和增殖情况优于普通胶原支架组(P 〈0.01)。表明负载胰岛素样生长因子1的壳聚糖胶原支架可显著促进人牙周膜细胞的增殖。  相似文献   

10.
背景:利用载体或缓释系统负载生长因子,既能保护生长因子的生物活性,又可以使生长因子缓慢释放,从而持续促进细胞生长及组织修复再生,是目前控制释放载体材料应用研究的方向之一.目的:观察血管内皮生长因子/纳米晶胶原基骨缓释系统的体外缓释效果及与种子细胞的生物相容性.方法:人骨髓间充质干细胞经体外诱导培养、扩增后种植于血管内皮生长因子+肝素/纤维连接蛋白+纳米晶胶原基骨支架(实验组)和单纯的纳米晶胶原基骨支架(对照组)行体外培养.测定缓释支架材料上血管内皮生长因子的释放量和持续时间;种植细胞后细胞的黏附率;培养3,7,10,14 d时支架材料中细胞数、碱性磷酸酶活性以及细胞在材料上的生长状况.结果与结论:①该缓释支架具有一定的血管内皮生长因子缓释效果,持续时间可达14 d.②第3代人骨髓间充质干细胞经成骨诱导培养,可表达成骨细胞表型:碱性磷酸酶细胞化学染色、Ⅰ型胶原免疫荧光染色均为阳性.③体外实验显示该缓释支架与入骨髓间充质干细胞具有优良的生物相容性:相同时间点实验组的细胞黏附率、支架上细胞数量及细胞碱性磷酸酶活性均明显高于对照组;扫描电镜发现两组材料上均有细胞生长,但实验组的细胞生长状况明显好于对照组.  相似文献   

11.
BACKGROUND: Lipoprotein(a) [Lp(a)] is a recognized pathogenic particle in human plasma, but its presence in the cerebrospinal fluid and its possible role in the central nervous system have not been documented. We tested the hypothesis that apolipoprotein(a) [apo(a)], free or as a component of the Lp(a) particle, can cross the blood-cerebrospinal fluid barrier and be found in the cerebrospinal fluid of patients affected by neurologic pathologies. METHODS: We studied paired cerebrospinal fluid/serum samples from 77 patients with inflammatory (n=20) or noninflammatory (n=34) blood-cerebrospinal fluid barrier dysfunction and without blood-cerebrospinal fluid barrier dysfunction (n=23). We used ELISA to measure Lp(a) concentrations and Western blot and immunodetection to analyze apo(a) isoforms in native and reducing conditions. RESULTS: Entire Lp(a) with either small or large apo(a) isoforms was present in the cerebrospinal fluid of patients with blood-cerebrospinal fluid barrier dysfunction, regardless of its pathogenesis. Multiple linear regression analysis showed that both serum Lp(a) concentration (P=0.003) and cerebrospinal fluid/serum albumin ratio (P<0.001) were predictors of the Lp(a) concentration in cerebrospinal fluid. CONCLUSIONS: Our results demonstrate that Lp(a) can cross a dysfunctional blood-cerebrospinal fluid barrier. The unusual presence of Lp(a) in the cerebrospinal fluid could extend some of its known pathogenic effects to the central nervous system.  相似文献   

12.
AIM: To study the therapeutical efficiency of plasmapheresis (PA) in patients with drug-resistant cardiac arrhythmias (CA) and its mechanisms. MATERIALS AND METHODS: Discrete PA sessions were carried out in 56 patients with drug-resistant CA: paroxysmal atrial arrhythmia (AA), ventricular and supraventricular premature contractions, supraventricular tachycardia of various etiology. Biochemical blood values, coagulographic parameters, lipid peroxidation (LPO), the spectrum of nonesterified fatty acids (NEFA), the level of medium-sized molecules were determined, ECG monitoring, EchoCG, and left ventricular radioisotope computed tomography were performed before and after a PA session. RESULTS: PA was effective in 50% of cases. The duration of its effect averaged 3.0 (1.25-5.0) months. PA was more beneficial for patients with IHD, AA with normal left atrial dimensions, and hyperlipidemia. The duration of the effect was significantly higher when antiarrhythmic drug therapy was continued after PA. Due to PA, there were significant decreases in the blood concentrations of cholesterol, medium-sized molecules, malonic dialdehyde (MDA) and in the proportion of polyunsaturated NEFA. The antiarrhythmic effect was associated with the decreases in MDA and NEFA, with a tendency for a reduction in the rate of chemiluminescence. CONCLUSION: PA may be used in the treatment of drug-resistant CA. The most significant mechanism of its antiarrhythmic activity is to recover sensitivity to antiarrhythmics. The intrinsic antiarrhythmic activity may be associated with its effect on NEFA metabolism and LPO; however, its mechanisms await further studies.  相似文献   

13.
背景:有研究表明线粒体基因变异与长寿相关,但在中国南方群体中线粒体单倍组D及其亚组与长寿的相关性研究至今少有报道,并且线粒体单倍组D及其亚组与载脂蛋白E基因相互作用对长寿的影响未见报道.目的:探讨在中国南方群体中线粒体单倍组D及其亚组与长寿的相关性,以及线粒体单倍组D及其亚组与载脂蛋白E基因的相互作用.方法:于2007/2008在广西壮族自治区巴马县选取对象1 038人,包括长寿组367人(年龄>90岁)和与长寿组无亲缘关系的当地健康对照组671人(年龄40~60岁).采用多聚酶链反应-限制性片段长度多态性技术检测线粒体单倍组D及其亚组(D4和D4a)的分布情况,并通过分层分析调整性别和载脂蛋白E基因混杂因素的作用.结果与结论:线粒体单倍组D及其亚组与长寿无明显相关性(P>0.05).按照性别进行分层分析后,在女性和男性群体中仍未发现线粒体单倍组D及其亚组与长寿的关联(P>0.05).然而,对载脂蛋白E基因分层分析后,在载脂蛋白E 携带者中长寿组和对照组线粒体单倍组D的分布频率差异有显著性意义(P<0.05).结果提示线粒体单倍组D可能是与载脂蛋白E基因的相互作用对长寿产生影响.  相似文献   

14.
目的:研究非选择性环氧化酶抑制剂舒林酸对结肠癌细胞株HT-29生长的影响及其参与引起细胞凋亡的可能机制。方法:采用四甲基偶氮唑蓝比色法检测舒林酸对结肠癌细胞增殖的抑制作用,采用激光共聚焦显微镜(LSM)观察细胞凋亡情况,采用流式细胞仪(FCM)分析其对细胞凋亡及细胞周期的影响。结果:舒林酸能呈剂量和浓度依赖性押制HT-29的增殖.在4.8mmol·L^-1时其抑制率可迭91.8%。AnnexinV/H染色后LSM观察到凋亡细胞胞膜绿染,胞核红染或呈桔黄色。FCM显示此药能促进细胞的凋亡,使处于G0/G1期的细胞比例显著降低。结论:舒林酸可抑制结肠癌细胞株HT-29生长,其机制可能与其阻止细胞周期的进展,诱导细胞凋亡有关。  相似文献   

15.
目的制备特发性血小板减少性紫癜(ITP)患者血浆IgG及其酶切片段,探讨其与血小板GPⅡb/Ⅲa和(或)GPⅠb/Ⅸ结合的免疫活性及其对正常人血小板聚集功能的影响。方法用改良MAIPA法和比浊法血小板聚集试验筛选出自身抗体阳性并且能抑制血小板聚集的患者,用蛋白A柱纯化其血浆IgG抗体并用胃蛋白酶制备F(ab’)2片段,改良单克隆抗体俘获血小板抗原技术(MAIPA)检测完整抗体及其酶切片段与血小板膜糖蛋白的结合活性;比浊法血小板聚集试验对比观察ITP患者血浆、纯化的IgG抗体及其酶切片段对正常人血小板聚集功能的影响。结果①68例慢性ITP患者中,34例(53.6%)血浆中抗GPⅡb/Ⅲa和(或)GPⅠb/Ⅸ自身抗体阳性,其中5例(14.7%)明显抑制了二磷酸腺苷(ADP)或瑞斯托霉素对血小板聚集的诱导作用;②用蛋白A柱结合蛋白酶酶切成功获得了纯化的IgG及F(ab’)2片段;③患者纯化的IgG及F(ab’)2片段均具有抗GPⅡb/Ⅲg或GPⅠb/Ⅸ活性,但去除IgG的血浆丧失了与GPⅡb/Ⅲa或GPⅠb/Ⅸ的结合活性;④2例患者纯化的IgG及其F(ab')2片段抑制ADP诱导的血小板聚集。结论F(ab’)2片段是IgG自身抗体的功能片段,它不但保留了良好的抗原结合活性并可抑制血小板聚集功能,其抑制聚集作用呈剂量依赖性。  相似文献   

16.
OBJECTIVE: To develop a simplified Tai Chi exercise program for frail older adults. DESIGN: For phase I, using a focus group, 40 frail Taiwanese older adults were interviewed to explore their viewpoints on Tai Chi and have been reported elsewhere. This paper emphasized on the phase II of the study in which the older adults' perspectives were validated by 10 experts using an evaluation survey. SETTING: Long-term care facilities. RESULTS: The newly developed simplified Tai Chi exercise program (STEP) included three stages-(1) warm-up: comprised nine exercises specifically designed to loosen up the body from head to toe; (2) Tai Chi movements: encompassed 12 easy-to-learn and easy-to-perform movements; (3) cool-down: included three activities to cease the chi and rest the body. CONCLUSIONS: The STEP should be further evaluated for its effectiveness in enhancing the relative well being and quality of life of frail older adults and its applicability as a floor activity in long-term care facilities.  相似文献   

17.
BACKGROUND: Holter monitoring is routinely used in patients referred for the evaluation of syncope, but its diagnostic value in different patient groups is unclear, as is its impact on device implantation (pacemaker or cardioverter-defibrillator). AIM: To determine the diagnostic yield of Holter monitoring in the routine evaluation of syncope, and its impact on subsequent device implantation. DESIGN: Retrospective record review. METHODS: We reviewed all Holter studies in patients referred with syncope between 2000 and 2005. Strict criteria were applied to determine whether a study was diagnostic. The diagnostic value of Holter monitoring (overall and in five subgroups: age, gender, structural heart disease, ejection fraction, medication) and its impact on the implantation of devices, were determined. RESULTS: Of 4877 Holter studies, 826 were performed in patients with syncope (age 72 +/- 15 years): 71 (8.6%) were considered to explain the syncope. Structural heart disease, ejection fraction and age were significant predictors of a diagnostic study (all p < 0.01), whereas gender and cardiac medication were not. A device was implanted in 33 patients (4.4%) whose initial Holter did not explain their syncope, after mean 7 months, whereas 45 patients (5.4%) received a pacemaker based on the Holter results (p = 0.32). DISCUSSION: The overall diagnostic yield of Holter monitoring in the evaluation of syncope was 8.6%, with dramatic differences between subgroups. Our data suggest that the impact of Holter monitoring on device implantation is generally overestimated.  相似文献   

18.
Fatigue after stroke   总被引:9,自引:0,他引:9  
OBJECTIVE: To determine the frequency and outcome of fatigue, its impact on functioning, and its relationship with depression in patients 3 to 13 months poststroke. DESIGN: Survey. SETTING: Community. PARTICIPANTS: Eighty-eight individuals from a pool of 181 consecutive patients previously admitted to an acute stroke service who were willing and able to complete the self-report questionnaires, and 56 elderly controls living independently in the community. MAIN OUTCOME MEASURES: Fatigue Impact Scale (a self-report measure of the presence and severity of fatigue and its impact on cognitive, physical, and psychosocial functions) and the Geriatric Depression Scale. RESULTS: The frequency of self-reported fatigue problems was greater in the stroke group (68%) than in the control group (36%, p < .001) and was not related to time poststroke, stroke severity, or lesion location. Forty percent of the stroke group reported that fatigue was either their worst or one of their worst symptoms. Patients attributed more functional limitations to their fatigue than did control subjects with fatigue. Although the presence of fatigue was independent of depression, the impact of fatigue on functional abilities was strongly influenced by depression. CONCLUSION: Fatigue can contribute to functional impairment up to 13 months after stroke, and its recognition and treatment are important for maximizing recovery.  相似文献   

19.
What is known and background: Although paracetamol (acetaminophen), N‐(4‐Hydroxyphenyl)acetamide, is one of the world’s most widely used analgesics, the mechanism by which it produces its analgesic effect is largely unknown. This lack is relevant because: (i) optimal pain treatment matches the analgesic mechanism to the (patho)physiology of the pain and (ii) modern drug discovery relies on an appropriate screening assay. Objective: To review the clinical profile and preclinical studies of paracetamol as means of gaining insight into its mechanism of analgesic action. Methods: A literature search was conducted of clinical and preclinical literature and the information obtained was organized and reviewed from the perspective of its contribution to an understanding of the mechanism of analgesic action of paracetamol. Results: Paracetamol’s broad spectrum of analgesic and other pharmacological actions is presented, along with its multiple postulated mechanism(s) of action. No one mechanism has been definitively shown to account for its analgesic activity. What is new and conclusion: Further research is needed to uncover the mechanism of analgesic action of paracetamol. The lack of this knowledge affects optimal clinical use and impedes drug discovery efforts.  相似文献   

20.
OBJECTIVE: Intratesticular varicocele (ITV) is an uncommon sonographic finding with controversial data concerning its prevalence and physiopathologic characteristics. The goal of this study was to determine the prevalence of ITV in a urogenital imaging department and to describe its sonographic features. METHODS: All identified cases of ITV were prospectively collected in the same imaging department. RESULTS: Intratesticular dilated veins (>2 mm) with a positive response to the Valsalva maneuver were referred to as ITV. Nine cases of ITV were detected in 8 patients (mean age, 60 years; range, 30-85 years) in a series of 1832 scrotal sonographic examinations performed over 5 years (0.4%). A history of homolateral scrotal surgery was found in 5 cases. In most cases, ITV was left sided (6/9) and located in the mediastinum testis (6/9) with associated extratesticular varicocele (8/9) and testicular hypotrophy (7/9). Five of the 7 hypotrophic testes had other causes of hypotrophy. CONCLUSIONS: Although variations do exist in the sonographic appearance of ITV, its specific sonographic and Doppler appearance should enable the radiologist to obviate further study. Intratesticular varicocele is often associated with ipsilateral testicular atrophy, but whether it is a cause or a consequence of testicular atrophy remains unclear.  相似文献   

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