LC-MS/MS法同时测定Beagle犬血浆中柚皮苷和新橙皮苷的含量

目的:建立同时测定Beagle犬血浆中柚皮苷和新橙皮苷的LC-MS/MS分析方法。方法:以芦丁为内标,血浆样品经乙腈沉淀蛋白处理,采用Diamonsil C18(150 mm×4.6 mm,5μm)色谱柱,以乙腈-0.1%甲酸水(25:75)为流动相,流速为0.4 ml/min,柱温20℃;采用ESI(+)离子源,MRM扫描检测离子对为柚皮苷m/z 581→273、新橙皮苷m/z 611→303、芦丁(内标)m/z611→303。结果:血浆中柚皮苷在0.025～2.5μg/ml、新橙皮苷在0.015～1.5μg/ml浓度范围内线性关系良好,二者的方法回收率均近100%,分析方法的日内日间精密度RSD值均小于10%,内源性物质不干扰样品测定。结论:本方法简便、准确、灵敏度高、专属性好,适用于Beagle犬血浆中柚皮苷和新橙皮苷测定及其体内药代动力学的研究。     

LC-MS/MS法测定大鼠血浆中芍药内酯苷的浓度及其在毒代动力学研究中的应用

目的 建立测定大鼠血浆中芍药内酯苷的液相色谱-串联质谱(LC-MS/MS)法.方法 血浆样品经乙酸乙酯-异丙醇(95:5,V/V)提取后,以Poroshell 120 EC-C18柱(50 mm×2.1 mm,2.7μm)为分析柱,乙腈-水为流动相梯度洗脱,采用ESI源在多反应监测(MRM)方式下进行正离子检测.用于定量分析的离子反应为m/z 498.5→m/z 197.1(芍药内酯苷)和m/z 251.3→m/z 108.2(拉科酰胺,内标).结果 芍药内酯苷血浆浓度测定方法的线性范围为20~2000 ng/ml,日内、日间精密度RSD%均<15%,准确度(RE)在±8.06%之间.结论 该法适用于芍药内酯苷在大鼠体内的毒代动力学研究.     

HPLC-MS/MS法测定大鼠血浆中麻黄碱、伪麻黄碱浓度及药动学研究

[目的]建立同时检测大鼠血浆中麻黄碱、伪麻黄碱的高效液相色谱-质谱联用法(HPLC-MS/MS),并对大鼠口服三拗汤后的麻黄碱、伪麻黄碱进行药代动力学研究。[方法]血浆样品经碱化和乙酸乙酯液-液萃取,以盐酸苯丙醇胺为内标,乙腈-0.1%甲酸水(4∶96)为流动相,经Agilent Zorbax SB-C18柱(4.6 mm×150 mm,5μm)分离,采用电喷雾离子源(ESI),以多反应监测(MRM)模式进行正离子检测,定量分析的离子反应分别为:m/z 166.2→m/z 148.2(麻黄碱),m/z 166.2→m/z 148.2(伪麻黄碱)和m/z 152.1→m/z 134.1(盐酸苯丙醇胺)。[结果]麻黄碱和伪麻黄碱血药浓度在20~10 000μg/L范围内线性关系良好,批内、批间精密度RSD均小于5.3%,高、中、低3种浓度平均方法回收率大于64.7%。[结论]该方法专属、快速、灵敏,可用于麻黄碱、伪麻黄碱的药代动力学研究。     

LC-MS/MS测定大鼠血浆中的野漆树苷及其药代动力学研究

目的：建立大鼠血浆中野漆树苷的LC-MS/MS测定方法,并研究静脉注射后野漆树苷在大鼠体内的药代动力学特征。方法大鼠血浆样品经乙酸乙酯萃取处理,用色谱柱Phenomenex C8（30 mm×2.00 mm,3μm）进行分离,以甲醇-水为流动相梯度洗脱,采用电喷雾离子源（ESI）,负离子模式,多反应监测（MRM）,选择离子分别为野漆树苷（m/z 577.6→269.1）,内标柚皮素（m/z 271.0→151.0）,将建立的方法应用于测定大鼠静脉注射野漆树苷后血浆的药物浓度,并采用DAS 2.0计算得主要药代动力学参数。结果大鼠血浆中野漆树苷在1～2000μg/L浓度范围内线性关系良好,日内、日间精密度（RSD）均＜10%,准确度（RE）在±7%之间,低、中、高3个浓度下提取回收率为86.8%～91.0%,无明显基质效应且稳定性良好。大鼠静注该药（5 mg/kg）后AUC0-t为（72627.8±18067.9）μg·min/L,t1/2为（52.1±14.3）min,CLz为（0.07±0.02）L/（min·kg）。结论该检测方法简单、快速、灵敏、准确,适用于野漆树苷血药浓度监测及其药动学研究。药代动力学参数表明大鼠静注该药后体内消除较快。     

液相色谱-质谱法测定孕妇尿液中3种残留的氯霉素类抗生素

目的 建立液相色谱-质谱法测定孕妇尿液中3种残留的氯霉素类抗生素,并进行方法学验证。方法 尿液利用Oasis HLB固相萃取小柱进行净化和富集,液相分离采用SHISEIDO MG-C18(100 mm×3.0 mm,3.0μm)色谱柱,流动相为乙腈-水（含0.1%甲酸）,梯度洗脱。质谱采用电喷雾离子源,选择离子监测模式检测[M-H]-：氯霉素m/z 320.8、氟苯尼考m/z 355.8、甲砜霉素m/z 353.8、甲苯磺丁脲（内标）m/z 268.9。结果 尿液中3种残留的氯霉素类抗生素在5～500 ng/mL范围内呈良好的线性关系（均r>0.999）;批内和批间的精密度及准确度良好（RSD<8.9%,∣相对误差（RE）∣<9.3%）;提取回收率为71.4%～108.5%,稳定性良好（∣RE∣<11.8%）;几乎没有残留;稀释不影响测定的精密度和准确度。共检测了18份不同孕期（孕早期、孕中期和孕晚期）孕妇的尿样,大部分样品能检测到氯霉素（但浓度低于最低定量限5 ng/mL）,其中少量样品能测出氟苯尼考,完全没有检测到甲砜霉素残留。结论 该方法操作简便、重现性好...     

固相萃取LC-MS/MS法测定疏血通注射液中次黄嘌呤浓度

[目的]建立固相萃取液相色谱串联质谱法(LC-MS/MS)测定疏血通注射液中次黄嘌呤浓度。[方法]疏血通注射液经固相萃取柱处理后,采用LC-MS/MS进行含量测定。色谱柱为Acquity BEH C18分析柱(100 mm×2.1 mm,1.7μm),以甲醇-水(含0.1%甲酸)为流动相进行梯度洗脱,流速0.3 mL/min。于ESI负离子模式下,以多反应离子监测(MRM)扫描方式进行监测,用于定量分析的离子为m/z 134.9→92.0(次黄嘌呤)和m/z 151.0→92.1(内标6-巯基嘌呤)。[结果]次黄嘌呤在质量浓度0.1～0.5μg/mL范围内与待测物和内标的峰面积比值呈良好线性关系(r=0.999 8),平均加样回收率为98.7%,RSD为0.31%。[结论]该方法简便、准确、灵敏,可用于疏血通注射液中次黄嘌呤的浓度测定。     

LC-MS/MS法测定人血浆中洛伐他汀及其活性代谢物的含量

目的 建立高效液相色谱-串联质谱法(LC-MS/MS)测定人血浆中洛伐他汀及其活性代谢物洛伐他汀酸的浓度.方法 样品用乙酸乙酯∶二氯甲烷(体积比为1∶1)提取浓集后进样,色谱柱为Phenomenex Gemini C18(50×3 mm, 3 μm),流动相为乙腈-水(85∶15).质谱采用ESI离子源MRM模式检测,以辛伐他汀为内标,按内标法定量. 洛伐他汀、洛伐他汀酸和内标的检测离子对分别为质荷比(m/z)427.4→325.4、445.4→343.4和436.4→325.4.结果 洛伐他汀在0.031～64 μg/L范围内线性关系良好(r=0.9987),最低定量限为0.031 μg/L,方法 回收率在96%～102%之间,日内、日间精密度相对标准偏差(RSD)分别小于5.3%及9.3%.结论 本文建立的方法 准确、灵敏,可用于洛伐他汀血药浓度测定及药代动力学研究.     

荭草花提取物的药代动力学研究

【目的】 建立血浆中原儿茶酸、山奈素-葡萄糖苷、槲皮苷和山奈素-鼠李糖苷4个指标成分的超高效液相色谱-串联质谱(UPLC-ESI-MS/MS)分析方法,采用DAS 2.0软件计算药动学参数,研究大鼠口服及静注给予荭草花提取物后的药代动力学特征。 【方法】 利用大鼠口服及静注给予荭草花提取物后,于不同时间点取血测定,血浆样品选择酸化后甲醇沉淀蛋白,UPLC-ESI-MS/MS检测。采用Waters BEH C18(2.1 mm×50 mm,1.7 μm)柱,流速: 0.35 mL/min,流动相: 0.1%甲酸乙腈-0.1%甲酸水梯度洗脱,口服给药的扫描方式为选择离子监测(SIR),用于定量分析的原儿茶酸、山柰素-葡萄糖苷、槲皮苷、山柰素-鼠李糖苷、葛根素(内标)监测离子分别为m/z 153.0;m/z 449.3;m/z 449.2;m/z 433.2;m/z 417.2。 静脉注射给药的扫描方式为为多反应离子监测(MRM),用于定量分析的原儿茶酸、山柰素-葡萄糖苷、槲皮苷、山柰素-鼠李糖苷、葛根素(内标)监测离子分别为m/z 153.0→109.0;m/z 449.3→287.1;m/z 449.2→303.1;m/z 433.2→287.1;m/z 417.2→267.1。 【结果】 建立的生物样品中各指标成分的分析方法符合样品测定要求。大鼠口服荭草花提取物后,原儿茶酸等4个指标成分能够较快吸收进入体内,T_max为0.5~0.7 h,MRT_0-t为3.3~13.5 h、AUC(0-t)为105.2~1327.3 μg/(L·h)、t_1/2z为8.3~15.9 h;大鼠静注荭草花提取物后,原儿茶酸等4个指标成分在大鼠体内消除较快,MRT_0-t为43.0~60.0 min、AUC(0-t)为56.7~379.8 mg/(L·min)、t_1/2z为41.9~108.2 min;原儿茶酸、山奈素-葡萄糖苷、槲皮苷和山奈素-鼠李糖苷在大鼠体内的绝对生物利用度分别为2.5%、0.17%、0.42%、0.30%。 【结论】 建立了荭草花提取物中原儿茶酸等主要成分在生物样品中超高效液相-质谱分析方法,方法学考察结果表明所建立的方法特异、快速、准确、灵敏。实验获得了原儿茶酸等4个指标成分在大鼠体内的药动学参数及口服生物利用度,为以荭草花为原料的药物制剂和药用资源深度开发奠定了科学依据。     

液相色谱-质谱联用法测定孕妇尿液中三种残留的氯霉素类抗生素

目的：建立液相色谱-质谱联用法测定孕妇尿液中3种残留的氯霉素类抗生素,并进行方法学验证。方法：尿液利用Oasis HLB固相萃取小柱进行净化和富集,液相分离采用SHISEIDO MG-C18(100mm×3.0mm,3.0μm)色谱柱,流动相为乙腈-水（含0.1%甲酸）,梯度洗脱。质谱采用ESI源,选择离子监测模式检测[M-H]-：氯霉素m/z 320.8、氟苯尼考m/z 355.8、甲砜霉素m/z 353.8、甲苯磺丁脲(内标) m/z 268.9。结果：尿液中3种残留的氯霉素类抗生素在5～500ng/mL范围内呈良好的线性关系（r均＞0.999）;批内和批间的精密度和准确度良好（RSD＜8.9% ,∣RE∣＜9.3%） ;提取回收率为71.4%～108.5%,稳定性良好（∣RE∣＜11.8%）;几乎没有残留;稀释不影响测定的精密度和准确度;共检测了18份来源不同的(孕早期、孕中期和孕晚期)孕妇尿样,大部分样品能检测到氯霉素（但低于最低定量限5ng/mL）,其中少量样品能测出氟苯尼考,完全没有检测到甲砜霉素残留。结论：该方法操作简便、重现性好,适用于尿液中3种氯霉素类抗生素的测定,可为氯霉素类抗生素的残留与孕妇不良妊娠结局的相关性研究提供方法学基础。     

基于尤斯灌流室技术的泽泻汤大鼠肠吸收研究

[目的]研究泽泻汤中成分白术内酯Ⅱ和Ⅲ在大鼠不同肠段的跨膜吸收特征。[方法]采用Agilent Zorbax Plus C18色谱柱,流动相为乙腈-0.1%甲酸水(42∶58),流速:0.3 mL/min。质谱电喷雾离子源,阳离子MRM模式,白术内酯Ⅱ和Ⅲ的定量离子分别为m/z233.2→m/z187,m/z249.15→m/z231。尤斯灌流室法考察泽泻汤在大鼠离体十二指肠、空肠、回肠、结肠的吸收,接收侧样品10 000 r/min离心15 min后用液相色谱-质谱色谱法(LC-MS/MS)分析。[结果]白术内酯Ⅱ在大鼠不同肠黏膜的表观渗透系数范围在6.10~25.22×10~(-6) cm/s之间,白术内酯Ⅲ在1.88~5.41×10~(-6)cm/s之间,两者最佳吸收部位是结肠。[结论]该方法专属性好,准确度高,可用于泽泻汤中白术内酯Ⅱ和Ⅲ的大鼠肠吸收研究。     

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.