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1.
目的探索零下非结冰(-0.8℃)保存C3A肝细胞的效果以及同细胞凋亡的关系。方法UW液(University of Wisconsin Solution)保存的C3A细胞分为以下2组:-0.8℃组,4℃组。低温保存24h、48h及72h后分别测定细胞存活率及凋亡率、AST释放、尿素合成功能及白蛋白分泌功能。结果零下非结冰(-0.8℃)72hC3A细胞存活率(86.49±2.80)%高于4℃(77.83±3.40)%,P〈0.001;细胞凋亡率(1.26±0.84)%低于4℃(9.16±1.99%,P〈0.001;AST释放(5.30±0.42)U/L低于4℃(8.18±1.05)U/L;细胞尿素合成功能(0.68±0.06)mmol/L优于4℃(0.40±0.07)mmol/L,P〈0.001;白蛋白分泌功能(2.06±0.22)mg/ml优于4℃(1.68±0.18)mg/ml(P=0.002)。结论0.8℃的UW液可明显提高低温保存C3A细胞存活率,有效地保护细胞尿素合成和白蛋白分泌功能。  相似文献   

2.
目的 比较UW液、Celsior液和HTK液4℃常规低温保存生物人工肝用L-02细胞的效果。方法 制备好的L-02细胞悬液分以下3组:UW液保存组(UW液组);Celsior液保存组(CS液组);HTK液保存组(HTK液组)。各组细胞于4℃低温保存72h后,分别测定细胞存活率及死亡率(流式细胞术),ALT及LDH释放,尿素合成功能及白蛋白分泌功能。结果 UW液较Celsior液和HTK液显著提高了4℃常规低温保存保存72h的L-02细胞的存活率[(60.05 ± 4.23)% vs (50.12 ± 3.99)%、(44.20 ±4.67)%],均P<0.05;降低了细胞死亡率[(39.95±4.23)%vs (49.88 ±3.99)%、(55.80%±4.67)%], 均P<0.05;抑制了ALT、LDH释放(均P <0.05);更好地维持了L-02细胞尿素合成功能[ (1.03 ± 0.23)mmol/L vs (0.80± 0.14)mmol/L、(0.48± 0.05)mmol/L]和白蛋白分泌功能[(8.36 ±1.38 )mg/L vs (6.41±1.25)mg/L、(5.19±0.41)mg/L), 均P<0.05。结论 同Celsior液和HTK液相比,使用UW液4℃常规低温保存L-02肝细胞可以明显的提高复温后细胞存活率,降低低温损伤引起的ALT、LDH释放,有效的保护肝细胞尿素合成功能和白蛋白分泌功能,但保存时间不应超过72h。  相似文献   

3.
林虎  李庆勇  蒋知新  张鋆歆  彭凌  杜平  张清华  高毅 《重庆医学》2011,40(28):2848-2850,2853
目的比较UW液、Celsior液和HTK液4℃常规低温保存生物人工肝用C3A细胞的效果。方法制备好的C3A细胞悬液分以下3组:UW液保存组(UW液组);Celsior液保存组(CS液组);HTK液保存组(HTK液组)。各组细胞于4℃低温保存72 h后,分别测定细胞存活率及死亡率(流式细胞术),丙氨酸氨基转移酶(ALT)及乳酸释放,尿素合成功能及清蛋白分泌功能。结果 UW液及Celsior液比HTK液显著提高了4℃低温保存72 h的C3A细胞的存活率(78.56%±4.67%vs 76.03%±3.53%,60.54%±3.18%,P<0.05);抑制了C3A细胞低温保存时ALT释放及乳酸的释放(P<0.05);更好地维持C3A细胞尿素合成功能[(0.52±0.11)mmol/L vs(0.51±0.06)mmol/L,(0.32±0.05)mmol/L,P<0.05]和清蛋白分泌功能[(1.79±0.26)g/L vs(1.75±0.21)g/L,(1.20±0.17)g/L,P<0.05]。UW液同Celsior液4℃低温保存C3A细胞的效果无差异。结论同HTK液相比,使用UW液或者Celsior液4℃保存C3A细胞可以明显的提高复温后细胞存活率,降低低温损伤引起的ALT释放和乳酸释放,有效的保护肝细胞尿素合成功能和清蛋白分泌功能。  相似文献   

4.
目的 建立一种操作简单、病变典型、稳定性好的大鼠急性坏死性胰腺炎模型.方法 100只SD大鼠采用随机数字法分为急性坏死性胰腺炎(acute necrotizing pancreatitis,ANP)模型组(A组, n=50)和对照组(C组,n=50),C组接受生理盐水对照,A组大鼠分3次腹腔内注射8% L-精氨酸(3×1.5 g/kg,每次间隔1 h),第3次注射后选取6、12、24、48 h和72 h 5个时相点观测两组大鼠胰腺病理学和血清淀粉酶水平的变化情况.结果 L-精氨酸注射后6 h可见胰腺少量炎细胞浸润、腺泡水肿;12 h见胰腺较多的炎细胞浸润,小叶结构紊乱和小叶间水肿;24 h 见胰腺大量炎细胞浸润,腺小叶结构损坏和局灶性坏死;48 h见弥漫性腺泡坏死和脂肪组织坏死;72 h见胰腺大片凝固性坏死和胰腺结构模糊;同时胰腺组织病理评分(A组vs C组)呈现相同的时相性变化[6 h (5.26±1.30)vs (0.67±0.52),P<0.01;12 h (6.83±1.20)vs(0.57±0.43),P<0.01;24 h (7.43±1.40) vs (0.47±0.63),P<0.01;48 h (8.27±1.10) vs(0.43±0.65),P<0.01;72 h(7.63±1.20) vs(0.67±0.52),P<0.01].各组血清淀粉酶水平(A组 vs C组),A组呈时相性变化,C组各时相点无明显差异[6 h (6 090±475)U/L vs (1 465±106)U/L, P<0.01;12 h (19 975±962)U/L vs (1 535±157)U/L, P<0.01;24 h(20 970±1 027)U/L vs(1 620±174)U/L, P<0.01;48 h (2 390±233)U/L vs (1 450±97)U/L, P<0.01;72 h(1 680±205)U/L vs (1 560±162)U/L,P>0.05].结论 分3次腹腔内注射大剂量L-精氨酸可成功制备操作简单、病变典型、重复性好和非侵入性的大鼠急性坏死性胰腺炎模型.  相似文献   

5.
目的探讨三磷酸肌醇(IP3)和Caspase3蛋白表达变化在genistein诱导肝癌细胞凋亡中的作用。方法以肝癌HepG2细胞培养72h为对照组,实验各组以60μmol/L的genistein作用于HepG2细胞不同时间后,应用同位素试剂盒检测细胞IP3含量,Westernblotting分析细胞Caspase3蛋白表达,流式细胞仪检测细胞凋亡率。结果Genistein作用于肝癌HepG2细胞12、24、48、72h,各时相IP3含量显著低于对照组[(12.0±1.4)pmol/106cells、(7.5±0.8)pmol/106cells、(5.6±0.5)pmol/106 cells、(4.3±0.6)pmol/106 cellsvs(29.2±0.6)pmol/106 cells,P<0.01];24h后Caspase3蛋白的RI显著高于对照组(2.7±0.2,7.4±0.5,7.4±0.5,30.7±1.6vs0.24±0.06,P<0.05);24h后各时相细胞凋亡率为显著高于对照组[(2.7±0.2)%、(7.4±0.5)%、(20.5±2.0)%、(30.7±1.6)%vs(2.6±0.1)%,P<0.01]。结论Genistein能减少IP3生成,上调Caspase3蛋白表达,诱导肝癌细胞凋亡。  相似文献   

6.
目的探讨胆红素对正常人肝细胞系L-02生长及凋亡的影响。方法将不同浓度的胆红素(终浓度分别为0、100、150、200、250、300 mg/L)作用于体外培养的正常人肝细胞L-02,分别通过光镜、Hoechst 33258荧光染色、四甲基偶氮唑蓝(MTT)法、DNA琼脂糖凝胶电泳、流式细胞仪Annexin-V和PI双标等技术观察不同浓度胆红素对L-02细胞的相对存活率的影响、细胞的凋亡梯度和凋亡率。结果经不同浓度胆红素溶液作用后,L-02细胞的增殖能力明显下降,其作用呈剂量依赖性;光镜观察显示凋亡细胞变圆,体积缩小;Hoechst 33258荧光强染;MTT法发现细胞生长受到抑制;经琼脂糖凝胶电泳发现200、250、300 mg/L胆红素溶液组可见到DNA梯形条带;流式细胞仪检测示各实验组细胞凋亡率为(1.24±0.72)%、(5.88±1.66)%、(19.69±3.52)%、(31.11±6.94)%、(57.3±8.71)%;均明显高于对照组细胞凋亡率(1.16±0.58)%(P<0.01)。结论胆红素可抑制L-02细胞生长并诱导其凋亡。  相似文献   

7.
目的:探讨冬凌草甲素(oridonin)对棕榈酸诱导的人脐静脉内皮细胞(HUVECs)损伤的保护作用。方法:体外培养HUVECs,噻唑蓝法(MTT)确定冬凌草甲素作用的最佳浓度和时间;细胞分为空白对照组、棕榈酸组、棕榈酸+oridonin预处理组(以下简称"oridonin预处理组",棕榈酸浓度为500μmol/L),oridonin预处理24h,棕榈酸作用48h。流式细胞仪检测细胞凋亡率,硝酸还原酶法检测各组细胞上清液中一氧化氮(NO)的浓度。结果:冬凌草甲素作用于HUVECs细胞的最佳时间是24h,最佳浓度为5μmol/L。与空白对照组比较,棕榈酸组HUVECs细胞凋亡率[(21.23±4.11)%vs(1.63±1.23)%,t=7.90,P<0.01]明显增加,上清液中NO的浓度[(114.67±17.01)μmol/Lvs(242.33±20.11)μmol/L,t=-8.396,P<0.01]明显降低。相对于棕榈酸组,oridonin预处理组HUVECs凋亡率明显降低[(7.77±2.57)%vs(21.23±4.11)%,t=-4.81,P<0.01],上清液中NO的浓度[(191.33±13.01)μmol/L vs(114.67±17.01)μmol/L,t=6.20,P<0.01]明显升高。结论:冬凌草甲素能抑制棕榈酸诱导的HUVECs细胞凋亡,并增加NO的浓度。  相似文献   

8.
目的:研究普鲁泊福(propofol)对肿瘤坏死因子α(TNF-α)诱导的小鼠脊髓神经元凋亡及Bax表达的影响.方法:脊髓神经元取自小鼠胎鼠,置于含B27的神经细胞培养基中培养,在培养第7天随机分为6组:对照组,50 μmol/L普鲁泊福组,TNF-α组,25 μmol/L普鲁泊福 TNF-α组,50 μmol/L普鲁泊福 TNF-α组,100 μmol/L普鲁泊福 TNF-α组.在相应组中加入不同终浓度的普鲁泊福,孵育30 min,再加入TNF-α至终末浓度为2 000 U/ml,培养24 h后,采用碘化丙锭(PI)/Hoechst33342双染法检测细胞凋亡,于荧光显微镜下观察凋亡细胞.采用免疫细胞化学的方法检测Bax的表达.结果:与对照组相比,TNF-α组细胞凋亡百分率明显升高[(21.8±1.1)% vs (2.8±0.8)%,P<0.01],Bax表达明显增加[(0.251±0.016) vs (0.141±0.015),P<0.01];而经不同浓度普鲁泊福(25,50,100 μmol/L)处理后再加入TNF-α的各组与TNF-α组相比,细胞凋亡百分率下降[(16.2±1.2)%、(15.3±0.6)%、(12.2±0.8)% vs (21.8±1.1)%,P<0.05、P<0.05、P<0.01],Bax的表达降低[(0.198±0.011)、(0.188±0.012)、(0.167±0.014) vs (0.251±0.016),P均<0.05],且呈现一定的剂量效应关系.结论:临床浓度的普鲁泊福可能通过调节Bax的表达进而抑制由TNF-α诱导的脊髓神经元凋亡.  相似文献   

9.
吡格列酮对内皮祖细胞增殖、凋亡及功能的影响   总被引:1,自引:0,他引:1  
目的:观察不同浓度吡格列酮对大鼠骨髓源性内皮祖细胞(EPCs)增殖、凋亡和功能的影响,并探讨其可能的作用机制.方法:正常SD大鼠24只,随机分为A组(对照组)和B,C,D组[吡格列酮组剂量依次为10,20,40 mg/(kg·d)],灌胃处理10 d后断颈处死,密度梯度离心法获取骨髓单个核细胞,经鉴定后证实为EPCs. 每组细胞培养4 d后消化、计数并用完全培养液培养. 24 h后检测NO生成量,3 d后检测凋亡情况,7 d后检测增殖能力. 结果:与A组相比,B,C,D组EPCs增殖能力明显增高[B,C,D组A490 nm分别为(0.423±0.004), (0.680±0.008), (0.443±0.005) vs A组A490 nm (0.143±0.006), P<0.01];凋亡程度降低[B,C,D组分别为(32.8±0.8)%, (30.9±2.3)%,(33.0±3.9)% vs A组(40.1±1.1)%, P<0.01];培养上清中NO浓度显著提高[B,C,D组分别为(29.2±3.7), (41.0±7.7), (28.7±6.4) μmol/L vs A组(18.9±1.4), P<0.05];与C组相比,B组和D组促进EPCs增殖能力弱(P<0.01),培养上清中NO浓度相对低(P<0.05). 结论:不同浓度吡格列酮均能提高EPCs数量和功能;20 mg/(kg·d)吡格列酮对EPCs的作用较强.  相似文献   

10.
李豫  张建福  张咏梅  刘美静  石玥  马小波 《医学争鸣》2008,29(24):2229-2231
目的:研究染料木黄酮后处理对人胃黏膜上皮细胞缺氧/复氧损伤的影响.方法:采用人胃黏膜上皮细胞系建立缺氧/复氧损伤模型.实验分为常氧对照组、缺氧/复氧组、染料木黄酮后处理组、DMSO溶剂对照组.各组分别于复氧2,4,8,16 h结束后用MTT法检测细胞存活率;并采用缺氧2h/复氧4 h模型做Hoechst 33258染色观察细胞凋亡情况.结果:MTT检测显示,缺氧/复氧组细胞存活率分别为(64.1±1.7)%,(53.7±1.9)%,(40.8±2.7)%,(33.6±0.9)%,与常氧对照组(100.0±0)%比较,细胞存活率明显降低,差异具有统计学意义(P<0.01).染料木黄酮后处理组(100 umol/L)细胞存活率分别为(80.5±1.3)%,(72.7±1.5)%,(71.3±1.2)%,(55.1±1.0)%,与缺氧/复氧组比较,细胞存活率明显升高,差异具有统计学意义(P<0.01).荧光染色显示,缺氧/复氧组可见较多核致密浓染的凋亡细胞,而染料木黄酮后处理组凋亡细胞明显减少.结论:染料木黄酮后处理通过抑制细胞凋亡对人胃黏膜上皮细胞缺氧/复氧损伤产生一定的保护作用.  相似文献   

11.
Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P < 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (>0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P <0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P < 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.  相似文献   

12.
Objective: To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods: SD rats served as both donors and recipients. 4℃ sodium lactate Ringer's was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava (IVC) inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results: Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion: This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.  相似文献   

13.
Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P< 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P< 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.  相似文献   

14.
目的:评价使用安心颗粒对急诊经皮冠状动脉介入术(PPCI)术后生活质量的影响.方法:将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组(术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次)和对照组(仅接受基础药物治疗).所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表(MIDAS)、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果:入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异(P>0.05);出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低(P<0.05);组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低(P<0.05).两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血(P>0.05).两组发生轻度血小板减少症的患者数比较无差异(P>0.05).结论:PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.  相似文献   

15.
Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P<0.01);the MAP decreased(P<0.01)and the HR increased drastically(P<0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.  相似文献   

16.
Objective:To investigate the gene expression of osteoprotegerin(OPG) and osteoclast differentiation factor(ODF) in the bone tissue of patients with hip fracture due to osteoporosis. Methods:OPGmRNA and ODFmRNA in the bone tissue in 50 cases of osteoporosis sufferers(over 50 years old) with hip fracture(Observer Group) and 30 cases of hip facture sufferers with no osteoporosis(Control group) were analyzed with the Semi-Quantitative RT-PCR method. Results:The mRNA expressed of ODF, OPG were both high in the patients with hip fracture. In the control group, the expression of OPG mRNA was observed, while the expression of ODF mRNA was very slight. Conclusion:Aged patients contained all signals including OPG, ODF that are essential for inducing osteoclastogenesis and promoting bone resorption.  相似文献   

17.
Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.  相似文献   

18.
Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.  相似文献   

19.
Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.  相似文献   

20.
A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care(~'4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes.  相似文献   

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