金属硫蛋白、同型半胱氨酸对血管内皮细胞的作用

目的:观察金属硫蛋白(metallothionein MT)对同型半胱氨酸(homocysteine Hcy)损伤血管内皮细胞的保护作用及机制。方法:培养血管内皮细胞,培养液中分别加入Hcy及Hcy+MT孵育血管内皮细胞,自动生化分析仪测培养液乳酸脱氢酶(LDH)漏出量,硫代巴比妥酸法测细胞丙二醛(MDA)含量,化学比色方法测超氧化物歧化酶(SOD)活力。结果:与对照组相比,Hcy组细胞LDH及蛋白漏出增加,细胞MDA含量明显升高(P<0.01);外源性给予MT(10-9mol/L,5×10-9mol/L,10-8mol/L)呈浓度依赖性地抑制Hcy引起的血管内皮细胞损伤,与Hcy组比,LDH漏出分别降低16.3%(P<0.05)、23.5%(P<0.01)和37.2%(P<0.01)。MDA含量分别减少15.8%(P<0.05)、21.1%(P<0.01)和36.8%(P<0.01)。结论:MT拮抗Hcy对血管内皮细胞的脂质过氧化损伤。     

同型半胱氨酸对家兔组织及血浆金属硫蛋白含量的影响

目的 :观察高同型半胱氨酸 (Hcy)血症时家兔组织及血浆金属硫蛋白 (MT)的变化。 方法 :家兔皮下注射Hcy复制高Hcy血症模型 ,测定肝、心、肾组织及血浆MT和丙二醛 (MDA)含量。 结果 :Hcy组家兔肝、心、肾组织及血浆MT水平较对照组分别升高 2 30 % (P <0 .0 1 )、1 30 % (P <0 .0 1 )、5 4 .8% (P <0 .0 1 )和 85 % (P <0 .0 1 ) ,而丙二醛水平则分别高 1 0 0 % (P <0 .0 1 )、71 % (P <0 .0 1 )、5 0 % (P <0 .0 1 )和 1 1 0 % (P <0 .0 1 )。注射Hcy同时用叶酸处理动物 ,肝、心和肾组织MT含量较Hcy组分别降低 36 % (P <0 .0 1 )、33% (P <0 .0 1 )、2 9%和 36 % (P <0 .0 1 ) ,丙二醛含量分别降低 5 2 % (P <0 .0 1 )、4 1 % (P <0 .0 1 ) 4 5 % (P <0 .0 1 )和 2 9% (P <0 .0 1 )。结论 :Hcy诱导家兔组织MT生成与氧化应激有关     

高同型半胱氨酸血症-动脉硬化家兔金属硫蛋白的变化

目的 :在高同型半胱氨酸 (Hcy)血症家兔动脉粥样硬化 (AS)模型上观察肝、心组织及血浆金属硫蛋白(MT)含量变化 ,认识AS时机体氧化防御系统的变化。方法 :家兔高蛋氨酸饮食 8周 ,复制高Hcy -动脉粥样硬化模型 ,分别以 [10 9Cd2 + ]血红蛋白饱和法和硫代巴比妥酸法测定家兔肝、心组织及血浆MT及丙二醛 (MDA)含量。结果 :AS家兔肝、心组织及血浆MT水平分别较对照组高 2 6 9% (P <0 .0 1)、4 6 % (P <0 .0 1)和 79% (P <0 .0 1)。肝、心组织及血浆MDA含量分别较对照组高 87% (P <0 .0 1)、83% (P <0 .0 1)和 4 7% (P <0 .0 1)。结论 :Hcy -AS家兔肝、心组织及血浆MT产生增多 ,可能与氧化损伤有关     

金属硫蛋白对阿霉素致心肌细胞氧化损伤的保护作用

目的 观察金属硫蛋白(metallothionein,MT)对阿霉素(adriamycin,ADR)致心肌细胞氧化损伤的保护作用.方法 将培养的心肌细胞随机分5组进行实验:①对照组;②阿霉素(1 mg/L)组;③阿霉素(1 mg/L)+MT1(5×10-9mol/L)组;④阿霉素(1 mg/L)+MT2(10-8mol/L)组;⑤阿霉素(1 mg/L)+MT3(5×10-8mol/L)组.MTT法计算心肌细胞存活率;试剂盒检测细胞培养液乳酸脱氢酶(LDH)漏出量、细胞丙二醛(MDA)和一氧化氮(NO)含量及超氧化物歧化酶(SOD)活力.结果 与对照组相比,阿霉素组心肌细胞存活率下降(P＜0.01),细胞LDH漏出量增加(P＜0.01),细胞MDA及NO含量升高(P＜0.01),细胞SOD活力降低(P＜0.01).分别应用5×10-9,10-8,5×10-8 mol/L MT与阿霉素共同处理心肌细胞,细胞存活率分别较单用阿霉素增加31.8%,47.8%和51.4%(均P＜0.01);细胞LDH漏出量分别较单用阿霉素降低20.5%,23.3%和34.6%(均P＜0.01);细胞MDA含量分别较单用阿霉素降低16.3%(P＜0.05),21.1%(P＜0.01),36.5%(P＜0.01);NO含量分别较单用阿霉素降低17.7%(P＜0.05),19.8%(P＜0.05)和30.1%(P＜0.01);心肌细胞SOD活性分别较单用阿霉素增加31.2%,36.5%和39.1%(均P＜0.01).结论 阿霉素导致心肌细胞氧化损伤,外源性MT具有降低这种心肌细胞氧化损伤的作用.     

HSP27对Hcy诱导的血管内皮细胞损伤的保护作用及其机制研究

目的 研究热休克蛋白27 (HSP27)对同型半胱氨酸(Hcy)诱导的血管内皮细胞损伤的保护作用及其机制.方法 培养人脐静脉内皮细胞,采用MTT法检测Hcy对人脐静脉内皮细胞(HUVECs)存活率的影响;Annexin V-FITC细胞凋亡检测试剂盒及流式细胞术检测Hcy及HSP27对细胞凋亡的影响;设计实验,采用总一氧化氮检测试剂盒(硝酸还原酶法)检测细胞培养液中NO水平、2',7'-二氯荧光黄双乙酸盐(DCFH-DA)活性氧(ROS)检测试剂盒及流式细胞术检测细胞内ROS来研究可能的保护机制.结果 HUVECs与不同浓度的Hcy作用后,0 mmol/L组存活率均值为97.33％,Hcy 0.2 mmol/L组存活率均值为95.17％,Hcy 0.4 mmol/L组存活率均值为90.72％,Hcy 0.6 mmol/L组存活率均值为78.29％,Hcy 0.8 mmol/L组存活率均值为63.65％,Hcy 1.0 mmol/L组存活率均值为41.51％.0 mmol/L组与Hcy 0.2 mmol/L组比较,差异无统计学意义(P＞0.05);Hcy 0.4、0.6、0.8、1.0 mmol/L组存活率均低于0 mmol/L组,差异有统计学意义(均P＜0.05).HSP27浓度越高,细胞的存活率越高,呈浓度依赖性,HSP27对HUVECs损伤具有保护作用.Hcy能够诱发内皮细胞凋亡,而HSP27能显著抑制这种凋亡,表明HSP27能减少Hcy对内皮细胞的损害.HSP27能抑制Hcy引起NO的减少和抑制Hcy诱导ROS的产生.结论 HSP27通过影响NO表达和调节细胞内ROS水平保护Hcy诱导的受损伤内皮细胞.     

Apelin-13通过AMPK通路抑制同型半胱氨酸诱导的内皮细胞损伤

目的 观察Apelin-13对同型半胱氨酸(Hcy)诱导的血管内皮细胞损伤的保护作用并探讨其可能的作用机制。方法采用10 mmol/L Hcy处理人脐静脉内皮细胞(HUVECs)24 h,诱导内皮细胞的损伤。采用不同浓度(0.1 μmol/L、1.0 μmol/L和10.0 μmol/L)的Apelin-13预处理1 h,再加入含Hcy的培养基中继续孵育24 h,观察细胞的损伤情况。采用MTT法测定细胞活力,比色法测定细胞培养液中乳酸脱氢酶(LDH)的活性。采用Western Blot检测HUVECs中磷酸化的腺苷酸活化蛋白激酶(AMPK)蛋白表达。结果与对照组比较,Hcy组细胞的存活率显著降低,细胞培养液中LDH活力显著性升高(均P＜0.05)。与Hcy组组比较,Apelin-13(1.0和10.0 μmol/L)预处理组细胞的存活率均显著升高,细胞培养液中LDH活力显著性降低(均P＜0.05)。Hcy组HUVECs细胞中p-AMPK蛋白的表达与对照组比较没有显著性差异(均P＞0.05)。与Hcy组比较,Apelin-13(1.0和10.0 μmol/L)预处理组HUVECs细胞中p-AMPK蛋白的表达显著性下调(均P<0.05)。AMPK抑制剂Compound C部分取消了Apelin-13的内皮细胞保护作用。结论Apelin-13抑制了同型半胱氨酸诱导的内皮细胞损伤,其机制与可能与上调AMPK的磷酸化有关。     

金属硫蛋白对同型半胱氨酸致血管内皮细胞损伤及脂质过氧化的影响

目的 观察金属硫蛋白（metallothionein，MT）拮抗同型半胱氨酸（homocysteine，Hcy）对血管内皮细胞损伤及脂质过氧化作用。方法培养人血管内皮细胞，台盼蓝排斥法计算细胞存活率，NADH氧化法测定乳酸脱氢酶（LDH）漏出量，^109Cd血红素饱和法测细胞MT含量，硫代巴比妥酸法测定细胞丙二醛（MDA）含量，化学比色方法测超氧化物歧化酶（SOD）及谷胱甘肽过氧化物酶（GSH-Px）活力。结果同型半胱氨酸降低血管内皮细胞存活率，增加细胞LDH漏出量，增加细胞MDA含量，降低细胞SOD、GSH-Px活力。外源性MT呈浓度依赖性地抑制Hcy引起的血管内皮细胞损伤及抗氧化酶活力降低；凝血酶诱导MT生成亦明显抑制Hcy引起的内皮细胞损伤和抗氧化酶损伤。结论Hey损伤血管内皮细胞，降低细胞抗氧化酶活力，外源性给予或内源性诱导MT生成均可拈抗Hcy的内皮细胞损伤及脂质过氧化作用。     

杏仁对家兔动脉粥样硬化及金属硫蛋白含量的影响

目的 :研究杏仁对家兔动脉粥样硬化 (AS)及肝、心组织、血浆金属硫蛋白 (MT)的影响 ,探讨杏仁对AS的防治作用及可能机制。方法 :高脂组家兔高脂饮食 8周 ,复制高脂血症 -动脉粥样硬化模型。杏仁组动物高脂饮食 4周后 ,腹腔注射杏仁液 ,对照组家兔喂养普通饲料 8周。以 [10 9Cd \〗血红素饱和法和硫代巴比妥酸法测定动物肝、心组织及血浆MT和MDA含量。结果 :AS组家兔肝、心组织及血浆MT水平分别较对照组升高 84% (P <0 0 5 )、48% (P <0 0 5 )、44 % (P <0 0 5 )。腹腔注射杏仁液后家兔肝、心组织和血浆MT含量进一步增高 ,分别较AS组升高 12 8% (P <0 0 5 )、3 3 % (P <0 0 5 )、46% (P <0 0 5 ) ,而MDA含量较AS组明显下降。结论 :杏仁可促使家兔组织MT合成增多 ,减轻AS的发生。     

哮喘对豚鼠金属硫蛋白合成的影响

目的 :观察哮喘豚鼠组织及血浆金属硫蛋白 (MT)含量的变化 ,认识哮喘时机体氧化防御系统的改变。方法 :复制豚鼠哮喘模型 ,分别以 [10 9Cd]-血红蛋白饱和法和硫代巴比妥酸法测定豚鼠组织及血浆MT及丙二醛(MDA)含量。结果 :哮喘豚鼠肝、心、肾和肺组织及血浆MT水平分别较对照组高 2 0 .3% (P <0 .0 5 )、6 4.3%(P <0 .0 5 )、5 2 .5 % (P <0 .0 1)、5 0 .0 % (P <0 .0 5 )、2 8.5 % (P <0 .0 5 )。MDA含量分别较对照组高 16 0 .8(P <0 .0 1)、38.5 % (P <0 .0 5 )、40 .0 % (P <0 .0 5 )、71.4% (P <0 .0 1)、89.5 % (P <0 .0 1)。结论 :哮喘豚鼠组织及血浆MT产生增多 ,并与氧化损伤有关     

低氧预适应对阿霉素致心肌细胞损伤的保护作用

目的观察低氧预适应(HPC)对阿霉素致心肌细胞损伤的保护作用。方法将培养的心肌细胞随机分为3组:对照组、阿霉素组、HPC+阿霉素组。MTT法测定心肌细胞存活率;试剂盒检测细胞培养液乳酸脱氢酶(LDH)漏出量;SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)和蛋白印迹(Western blot)半定量检测核转录因子-κB(NF-κB)蛋白表达量的改变。结果与对照组相比,阿霉素组心肌细胞存活率显著下降(P0.01),细胞LDH漏出量显著增加(P0.01),NF-κB蛋白表达水平增高显著(P0.01);与单纯阿霉素组相比,低氧预适应组心肌细胞存活率显著升高(P0.01),细胞LDH漏出量显著降低(P0.01),NF-κB蛋白表达水平明显降低(P0.01)。结论阿霉素导致心肌细胞氧化损伤,低氧预适应可能通过抑制NF-κB蛋白表达水平对其起保护作用。     

Value of D-Dimers in patients with acute aortic dissection

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.     

Research of combined liver-kidney transplantation model in rats

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.     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

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.     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

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.     

The comparative studies of the influences of Urapidil and Nicardipine on sino-atrial node function, atrio-ventricular node function and hemodynamics

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.     

Expression of OPGmRNA and ODFmRNA in the patients of hip fracture due to osteoporosis

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.     

The clinicopathological and immuohistochemical analysis of solid-pseudopapillary tumor of the pancreas： report of 9 cases

Objective：To investigate the clinical features, pathological characteristics and immunophenotype of solid-pseudopapillary tumor of the pancreas（SPTP）. Methods：Nine surgically treated cases of SPTP were retrospectively reviewed. Hematoxylin and Eosin（HE） staining and immunohistochemical staining were used to analyze all cases, and the general clinical data was collected. Results：Six patients were asymptomatic except for a palpable mass. Two patients complained of vague-epigastric pain. One patient appeared jaundice. The tumor was encapsulated and solid tissues alternately with cystic tissues. Histologically, the histological structure of solid portion was pseudopapillary with a fibrovascular core. Tumor cells were uniform and medium-sized which were arranged in sheets ets or nests or pseudopapillary patterns. Immunohistochemical studies demonstrated that SPTP proved positive in vimentin（9/9 cases）, AAT（9/9 cases）, NSE（9/9 cases）, ACT（7/9 cases）, CK20（2/9 cases）, CgA（1/9 cases）, S-100（3/gcases）, PR（4/gcases）, Syn（3/9 cases） and CD56（5/9cases）, negative in CEA and ER. Conclusion：SPTP is a tumor predominantly occurring in young women frequently without special symptoms. This tumor has various characteristical histological patterns with different immunophenotype.     

Dynamic Changes in Serum Estradiol and Progesterone levels in Patients of Premenstrual Syndrome with Adverse Flow of Liver-qi

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.     

Real-time Three-dimensional Echocardiography in Assessment of Left Ventricular and Right Ventricular Volumes

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.     

SCREENING FOR A 21-CHROMOSOME ABNORMALITY IN PREIMPLANTED EMBRYOS OF ELDERLY WOMEN

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.