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Antagonism of estrogen-mediated cell proliferation by raloxifene in prevention of ageing-related prostatic hyperplasia

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Asian Journal of Andrology 2010年第5期12卷 735-743页

作者： Rui yang yu-Xia Ma lin-feng Chen Ying Zhou Zhan-Po yang yan zhu Xiao-ling Du Jian-Dang Shi hong-Shun Ma Ju Zhang Bioactive Materials Key Lab of Ministry of Education Department of Biochemistry and Molecular Biology College of Life Sciences Nankai University Tianjin 300071 China Dana Farber Cancer Institute Harvard Medical School Boston MA 02445 USA Department of Urology Tianjin First Central Hospital Tianjin 300192 China Molecular Cardiology Research Institute Tufts Medical Center Tufts University School of Medicine Boston MA 02111 USA

Estrogen has important roles in the initiation and development of benign prostatic hyperplasia （BPH）. Regulators of the estrogen receptor （ER） are tissue- and cell-specific. We evaluated the effect of estrogen antagonist, raloxifene （Ral）, on the prevention and treatment of BPH by investigating its effect on the proliferation of two different prostate cell lines： a stromal cell line, WPMY-1, and a benign prostatic hyperplasia epithelial cell line, BPH-1. We additionally evaluated its effect on prostatic hyperplasia induced by estrogen and androgen in a rat model. The effect of Ral on the prevention of prostatic hyperplasia was analyzed by haematoxylin and eosin staining and quantitative immunohistochemistry （IHC） for proliferating cell nuclear antigen and α-smooth muscle actin. In vitro and in vivo, tamoxifen （Tam）, another anti-estrogen drug, and finasteride （Fin）, a drug for the clinical treatment of BPH, served as efficacy controls. The in vitro data showed that neither Ral nor Tam alone affected the proliferation of WPMY-1 and BPH-1, but both antagonized the effect of oestradiol in promoting the proliferation of the two cells. Results from the IHC staining of the rat prostates indicated that, similar to Tam and Fin, Ral inhibited the proliferation of stromal cells in vivo. Interestingly, in contrast to Tam, both Ral and Fin inhibited the proliferation of epithelial cells. Furthemore, Ral treatment much strongly decreased the number of prostatic acini and the surrounding layers of smooth muscle cells than Fin （P 〈 0.05）. Our data showed for the first time that Ral may have a role in the response of the rat prostate to selective ER modulators.

关键词： estrogen prostatic hyperplasia raloxifene rats

One-year status of hepatic venous pressure gradient measurement from 85 hospitals in China

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Portal Hypertension & Cirrhosis 2024年第2期3卷 116-123页

作者： Jitao Wang Ting Cui linpeng Zhang Zhiping yan Lei Li Jinjun Chen Junhui Sun Hua Xiang He zhu Hao Wu Jiancuo Gengzang feng Zhang Wei Wu Guohong Cao Wuhua Guo Haibin Shi Hui Xue Guangjun Huang Qiang yu Meng Niu Huiling Xiang Derun Kong Wei Gou Xuefeng Luo Fuliang He Min yuan Wei Liu Yong Wang hongjie Hu Xiaoli zhu Tao yu Qinxue Sun Wei Qu Ting Lu Deqiang Ma Li Zhang Jun Ma Jun yang Ketao Mu Xiaoliang zhu Kai Xiong Huiguo Ding Shengjuan Yao Mingsheng Huang Fang Wang Zexin Wang Yong Huang Jianan Li hongfeng Yi Birun Huang Zhongwei Zhao Duiping feng yanming Lei Changlong Hou Wenbo Guo Shirong Liu Gaojun Teng Kangshun zhu Fan Wu Xiaogang Hu Wenfeng Zhang Shaoqi yang Zhouchao Hu Pengfei Pang Shaowu zhuang Weidong Wang Qingliang zhu Kunpeng Zhang Chengyu Liu Dongliang Li Chao Ma hong Jiang Xingong Zhao Weixin Ren Zhiwei Wang Lei yu Fuhuang lin Chaoguang yan feng Wang Lei yu Hui Huan Wenyong Shen Shaofei He Chengda Zhang Jinlun Bao Jiangtao Liu Jun Wu Jianbo Zhao Jian Zhang Xiaolong Qi Hebei Provincial Key Laboratory of Cirrhosis&Portal Hypertension Xingtai People's HospitalXingtaiHebeiChina Center of Portal Hypertension Department of RadiologyZhongda HospitalSchool of MedicineSoutheast UniversityNanjingJiangsuChina Department of Interventional Therapy Beijing Shijitan Hospital Affiliated to Capital Medical UniversityBeijingChina Department of Intervention Taiyuan Third People's HospitalTaiyuanShanxiChina Department of Intervention Zhongshan Hospital Affiliated to Fudan UniversityShanghaiChina Department of Interventional Medicine The First Hospital of Lanzhou UniversityLanzhouGansuChina Department of Infectious Diseases Nanfang Hospital of Southern Medical UniversityGuangzhouGuangdongChina Hepatobiliary and Pancreatic Intervention Center The First Affiliated Hospital of Zhejiang University School of MedicineHangzhouZhejiangChina Interventional Vascular Surgery Hunan Provincial People's HospitalChangshaHunanChina Department of Intervention Sixth People's Hospital of ShenyangShenyangLiaoningChina Department of Gastroenterology West China Hospital of Sichuan UniversityChengduSichuanChina Department of Intervention The Fourth People's Hospital of Qinghai ProvinceXiningQinghaiChina Department of Gastroenterology Nanjing Drum Tower HospitalNanjingJiangsuChina Department of Gastroenterology The First Affiliated Hospital of Wenzhou Medical UniversityWenzhouZhejiangChina Department of Radiology Shulan(Hangzhou)HospitalHangzhouZhejiangChina Department of Intervention Mengchao Hepatobiliary Hospital of Fujian Medical UniversityFuzhouFujianChina Department of Interventional Radiology The First Affiliated Hospital of Nanjing Medical UniversityNanjingJiangsuChina Department of Gastroenterology The First Affiliated Hospital of Xi'an Jiaotong UniversityXi'anShaanxiChina Department of Gastroenterology Shandong Provincial Hospital Affiliated to Shandong First Medical UniversityJinanShandongChina Department of Interventional Radiology Fifth Medical Center

Aims:Surveys and research on the applications of the hepatic venous pressure gradient(HVPG)are important for understanding the current status and future development of this technology in *** article aimed to investigate the status of hepatic venous pressure gradient measurement in China in ***:We investigated the overall status of HVPG technology in China-including hospital distribution,hospital level,annual number of cases,catheters used,average cost,indications,and current challenges by using online *** counting the number and percentages of cases of these results,we hope to clarify the current status of HVPG measurements in ***:According to the survey,85 hospitals in China used HVPG technology in 2022 distributed across 29 provinces.A total of 4989 HVPG measurements were performed in all of the surveyed hospitals in 2022,of which 2813 cases(56.4%)were measured *** average cost of HVPG measurement was 5646.8±2327.9 *** the clinical teams who performed the measurements(sometimes multiple per hospital),94.3%(82/87)used the balloon method,and the majority of the teams(72.4%,63/87)used embolectomy ***:This survey clarified the clinical application status of HVPG in China and confirmed that some medical institutions in China have established a foundation for this *** is still necessary to continue promoting and popularizing this technology in the future.

关键词： application status hepatic vein pressure gradient questionnaire survey

Hypoxia-inducible factor-1α–mediated upregulation of CD99 promotes the proliferation of placental mesenchymal stem cells by regulating ERK1/2

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World Journal of Stem Cells 2021年第4期13卷 317-330页

作者： Xu-Dong feng Jia-Qi zhu Jia-Hang Zhou Fei-yan lin Bing feng Xiao-Wei Shi Qiao-ling Pan Jiong yu Lan-Juan Li hong-Cui Cao State Key Laboratory for the Diagnosis and Treatment of Infectious Diseases The First Affiliated HospitalZhejiang University School of MedicineHangzhou 310003Zhejiang ProvinceChina

BACKGROUND As human placenta-derived mesenchymal stem cells(hP-MSCs)exist in a physiologically hypoxic microenvironment,various studies have focused on the influence of ***,the underlying mechanisms remain to be further *** The aim was to reveal the possible mechanisms by which hypoxia enhances the proliferation of *** A hypoxic cell incubator(2.5%O2)was used to mimic a hypoxic *** counting kit-8 and 5-ethynyl-20-deoxyuridine incorporation assays were used to assay the proliferation of *** cell cycle was profiled by flow *** profiling of hP-MSCs under hypoxia was performed by RNA ***99 mRNA expression was assayed by reverse transcription-polymerase chain *** interfering RNA-mediated hypoxia-inducible factor 1α(HIF-1α)or CD99 knockdown of hP-MSCs,luciferase reporter assays,and the ERK1/2 signaling inhibitor PD98059 were used in the mechanistic *** expression was assayed by western blotting;immunofluorescence assays were conducted to evaluate changes in expression *** Hypoxia enhanced hP-MSC proliferation,increased the expression of cyclin E1,cyclin-dependent kinase 2,and cyclin A2,and decreased the expression of *** hypoxia,CD99 expression was increased by HIF-1α.CD99-specific small interfering RNA or the ERK1/2 signaling inhibitor PD98059 abrogated the hypoxia-induced increase in cell *** Hypoxia promoted hP-MSCs proliferation in a manner dependent on CD99 regulation of the MAPK/ERK signaling pathway in vitro.

关键词： Hypoxia-inducible factor 1α Hypoxia Mesenchymal stem cells Proliferation CD99 RNA sequencing assay MAPK/ERK signaling pathway

麝香追风止痛膏缓解膝骨关节炎(寒湿痹阻证)疼痛有效性与安全性的随机、双盲、阳性药平行对照、多中心临床试验

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中国中药杂志 2024年第9期49卷 2512-2521页

作者：丰芬方妤陈日高阎博华黄勇吴斌李成荫段华张林辉唐流刚尉迎丽汪国友王丽彬郭兵朱鸿飞姜维成武丽娜马林成都中医药大学附属医院四川成都610032 重庆市中医院重庆400011 四川省人民医院四川成都610072 四川省骨科医院四川成都610041 西南医科大学附属中医医院四川泸州646099 重庆市北碚区中医院重庆400799 重庆市永川区中医院重庆402105 泰州市姜堰中医院江苏泰州225599

麝香追风止痛膏具有祛风除湿,散寒止痛的功效,用于寒湿痹阻所致关节、肌肉疼痛,扭伤疼痛。为进一步评价麝香追风止痛膏缓解膝骨关节炎(寒湿痹阻证)疼痛的有效性与安全性,该研究开展了随机、双盲、阳性药物平行对照的多中心临床试验。对... 详细信息

麝香追风止痛膏具有祛风除湿,散寒止痛的功效,用于寒湿痹阻所致关节、肌肉疼痛,扭伤疼痛。为进一步评价麝香追风止痛膏缓解膝骨关节炎(寒湿痹阻证)疼痛的有效性与安全性,该研究开展了随机、双盲、阳性药物平行对照的多中心临床试验。对纳入240例受试者采用分层随机化方法,按1∶1随机分为试验组和对照组,2组中6 h贴敷和12 h贴敷各60例。试验组贴敷麝香追风止痛膏,对照组贴敷麝香追风膏,疗程均为21 d,在治疗后第7、14、21天进行随访。FAS分析显示,(1)主要疗效指标视觉模拟量表(VAS)评分方面,试验组12 h贴敷后与基线差值均数为3.02,对照组为2.31,试验组治疗后VAS评分下降程度优于对照组(P<0.05);试验组6 h贴敷后与基线差值均数为3.19,对照组为2.48,试验组治疗后VAS评分下降程度优于对照组(P<0.05)。(2)VAS评分疼痛缓解有效率方面,试验组12 h贴敷后有效率为93.22%,对照组为73.33%,试验组优于对照组(P<0.05);试验组6 h贴敷后有效率为88.33%,对照组为63.33%,试验组优于对照组(P<0.05)。结果表明,麝香追风止痛膏贴敷12、6 h均能有效缓解膝骨关节炎(寒湿痹阻证)患者疼痛,显著降低疼痛VAS评分,降低西蒙和马斯特大学关节炎指数(WOMAC)量表疼痛、僵硬、关节功能评分,临床主要疗效指标疼痛VAS评分的改善程度明显优于麝香追风膏,其缓解膝骨关节炎(寒湿痹阻证)疼痛疗效肯定。安全性方面,麝香追风止痛膏除导致个别患者用药部位皮肤局部出现中药贴膏剂连续使用常见的轻度过敏反应外,未见其他不良反应,也未见患者肝、肾功能与心电图等有异常影响,安全性较高,临床可继续推广运用。

关键词：麝香追风止痛膏膝骨关节炎临床试验安全性有效性

Microstructure and physical properties of Cu-2.3Ni-0.5Si alloy with thermo-mechanical treatment

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Rare Metals 2023年第1期42卷 64-69页

作者： Dong-Mei Liu Cong-Shuai Yin yan-hong Hua Dong-lin Zhang yu-Ao Li Bo-Jie Ma xiu-Chen Zhao Hao-feng Xie Li-Jun Peng Guo-Jie Huang Xu-Jun Mi State Key Laboratory of Nonferrous Metals and Processes GRINM Group Co.LtdBeijing100088China GRIMAT Engineering Institute Co. Ltd.Beijing101407China General Research Institute for Nonferrous Metals Beijing100088China School of Materials Science and Engineering Beijing Institute of TechnologyBeijing100081China

For Cu-Ni-Si alloys,cold rolling is usually performed after solution treatment to enhance physical properties such as strength and electrical conductivity during the aging *** paper reports the variation in microstructure and physical properties during the aging process of Cu-2.3Ni-0.5Si alloy cold-rolled at room temperature(RT) and cryogenic temperature(CT).When aged at 450℃ for 2 h,RT-rolled samples exhibited a maximum hardness of HV5234 with an electric conductivity of 39.9% *** CTrolled samples,the peak hardness was achieved when the samples were aged at 450% for 1 h.

关键词： alloy Microstructure microstructure

The molecular biology of pancreatic adenocarcinoma:translational challenges and clinical perspectives

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Signal Transduction and Targeted Therapy 2021年第8期6卷 2348-2370页

作者： Shun Wang yan Zheng feng yang Le zhu Xiao-Qiang zhu Zhe-Fang Wang Xiao-lin Wu Cheng-Hui Zhou Jia-yan yan Bei-yuan Hu Bo Kong De-Liang Fu Christiane Bruns yue Zhao Lun-xiu Qin Qiong-zhu Dong Department of General Surgery Huashan HospitalCancer Metastasis InstituteFudan UniversityShanghaiChina Department of Pancreatic Surgery Pancreatic Disease InstituteHuashan HospitalFudan UniversityShanghaiChina School of Biomedical Sciences Li Ka Shing Faculty of MedicineThe University of Hong KongHong KongChina General Visceral and Cancer SurgeryUniversity Hospital of CologneCologneGermany Department of Biliary-Pancreatic Surgery Renji HospitalSchool of MedicineShanghai Jiao Tong UniversityShanghaiChina Department of Surgery Klinikum rechts der IsarSchool of MedicineTechnical University of Munich(TUM)MunichGermany Key laboratory of whole-period monitoring and precise intervention of digestive cancer Shanghai Municipal Health Commission(SMHC)ShanghaiChina

Pancreatic cancer is an increasingly common cause of cancer mortality with a tight correspondence between disease mortality and ***,it is usually diagnosed at an advanced stage with a very dismal *** to the high heterogeneity,metabolic reprogramming,and dense stromal environment associated with pancreatic cancer,patients benefit little from current conventional *** insight into the biology and genetics of pancreatic cancer has supported its molecular classification,thus expanding clinical therapeutic *** this review,we summarize how the biological features of pancreatic cancer and its metabolic reprogramming as well as the tumor microenvironment regulate its development and *** further discuss potential biomarkers for pancreatic cancer diagnosis,prediction,and surveillance based on novel liquid *** also outline recent advances in defining pancreatic cancer subtypes and subtype-specific therapeutic responses and current preclinical therapeutic ***,we discuss prospects and challenges in the clinical development of pancreatic cancer therapeutics.

关键词： clinical mortality diagnosis

Titanium nitride nanorod array/carbon cloth as flexible integrated host for highly stable lithium-sulfur batteries

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Rare Metals 2023年第12期42卷 4115-4127页

作者： Rui-Qing Liu feng Jin Min Gu Dong-Wen Zhang Lu-Lu He Wen-xiu Liu Wen-feng zhu Kun Xie Jing-Yi Wu Yi-Ran Liu Wei-Wei yang xiu-Jing lin Li Shi Xiao-Miao feng Zhen Hou Ji-Gang Zhou yan-Wen Ma State Key Laboratory of Organic Electronics and Information Displays and Institute of Advanced Materials(IAM) Jiangsu National Synergetic Innovation Center for Advanced Materials(SICAM)Nanjing University of Posts and TelecommunicationsNanjing 210023China Department of Applied Physics The Hong Kong Polytechnic UniversityHung Hom 1000872Hong KongChina Canadian Light Source Inc. University of SaskatchewanSaskatoonSK S7N 2V3Canada Suzhou Vocational Institute of Industrial Technology Suzhou 215104China

Lithium-sulfur(Li-S)batteries have been regarded as promising energy-storage systems,due to their high theoretical capacity and energy ***,the carbonaceous sulfur hosts suffer from weak binding force between the hosts and polysulfides,restricting the cyclic stability of sulfur ***,the presence of binder and conductive agent in the traditional electrode reduces its energy *** study demonstrates that titanium nitride(TiN)nanorod array on carbon cloth(CC)is employed as a flexible host for highly stable Li-S batteries via solvothermal synthesis-nitridation *** the one hand,the flexible integrated network composed of three-dimensional TiN nanorod array and CC significantly improves the conductivity,increases the electron transport and electrolyte penetration of *** the other hand,the 3D structure of TiN/CC and the enhanced polarity of TiN effectively strengthen the physical and chemical double adsorption for *** a result,the combination of TiN nanorod array and CC synergistic ally promotes sulfur utilization and electrochemical performances of S@TiN/CC cathode.A discharge capacity of1015.2 mAh·g^(-1)at 0.5C after 250 cycles and 604.1mAh·g^(-1)at 3C after 250 cycles is *** a larger current density of 5C,the resulting S@TiN/CC cathode maintains a high discharge capacity of 666.6 mAh·g^(-1)and the Coulombic efficiency of about 100%.

关键词： Titanium nitride Nanorod array Integrated structure Chemical adsorption Lithium-sulfur batteries

巴戟天寡糖胶囊用于轻、中度抑郁症患者巩固期治疗的临床研究

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中国临床药理学杂志 2024年第6期40卷 815-819页

作者：周书喆韩祖成王秀珍陈燕清呼亚玲余雪芹王斌红樊国珍桑红海英贾智捷王占敏魏琰朱建国宋学勤刘志东况利王洪明田峰李玉欣张玲林海吴斌王超英刘畅孙佳凡闫少校刘君谢守付房茂胜宓为峰张鸿燕北京大学第六医院北京大学精神卫生研究所国家卫生健康委员会精神卫生学重点实验室(北京大学)国家精神心理疾病临床医学研究中心(北京大学第六医院)国家药物临床试验机构办公室北京100191 陕西省中医医院精神科陕西西安710003 沈阳市精神卫生中心精神科辽宁沈阳110168 山西中医学院附属医院精神科山西太原030024 山西大医院中医科山西大同037046 重庆市精神卫生中心精神科重庆400036 太原市精神病医院精神科山西太原030045 吉林省神经精神病医院精神科吉林松原138001 长春市第六医院精神科吉林长春130052 辽宁中医药大学附属医院精神科辽宁沈阳110033 河北省荣军医院精神科河北石家庄050899 衡水人民医院神经科河北衡水053099 郑州大学第一附属医院精神科河北郑州450052 天津市第四中心医院精神科天津300142 重庆医科大学附属第一医院精神科重庆400042 成都市第四人民医院精神科四川成都610036 山西医科大学第二医院精神科山西太原030001 河北省精神卫生中心精神科河北保定071030 首都医科大学附属北京安定医院精神科北京100120 西安市中医医院精神科陕西西安710021 西安市精神卫生中心精神科陕西西安710061 湖南省脑科医院精神科湖南长沙410021 吉林大学第一医院精神科吉林长春130061 山西省中西医结合医院精神科山西太原030013 北京回龙观医院精神科北京102208 辽宁中医药大学附属第二医院精神科辽宁沈阳110034 大连市第七人民医院精神科辽宁大连116086 武汉市精神卫生中心精神科湖北武汉430012

目的观察巴戟天寡糖胶囊用于治疗轻、中度抑郁症患者的24周巩固期的疗效和安全性。方法本研究采用开放、单臂、多中心设计,纳入已完成巴戟天寡糖急性期治疗的成人轻、中度抑郁症患者,继续给予巴戟天寡糖胶囊24周,巩固期维持治疗剂量不变... 详细信息

目的观察巴戟天寡糖胶囊用于治疗轻、中度抑郁症患者的24周巩固期的疗效和安全性。方法本研究采用开放、单臂、多中心设计,纳入已完成巴戟天寡糖急性期治疗的成人轻、中度抑郁症患者,继续给予巴戟天寡糖胶囊24周,巩固期维持治疗剂量不变,用汉密尔顿抑郁量表(HAMD-17)、汉密尔顿焦虑量表(HAMA)、临床总体印象量表-严重度(CGI-S)和亚利桑那性体验量表(ASEX)评估治疗前后的变化,评估患者的缓解率、复燃率及复燃时间,观察患者治疗相关不良事件的发生情况。结果治疗前后患者的HAMD-17评分分别为(6.60±1.87)和(5.85±4.18)分,HAMA评分分别为(6.36±3.02)和(4.93±3.09)分,CGI-S评分分别为(1.49±0.56)和(1.29±0.81)分,ASEX评分分别为(15.92±4.72)和(15.57±5.26)分,在统计学上差异均有统计学意义(均P<0.05)。用药24周末,患者的缓解率为54.59%(202例/370例),复燃率为6.49%(24例/370例),复燃时间为(64.67±42.47)d。巴戟天寡糖胶囊药物不良反应发生率为15.35%(64例/417例)。结论巴戟天寡糖胶囊能用于轻、中度抑郁症患者的巩固期治疗,且具有良好的耐受性和安全性。

关键词：巴戟天寡糖胶囊抑郁症疗效安全性

Clinical benefits of a modified Cryopiece system for cryopreservation of rare ejaculated and testicular spermatozoa for ICSI

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Asian Journal of Andrology 2022年第5期24卷 533-539页

作者： Wei Chen Chuan Huang Peng Li feng Liu Jian Sun Zi-Jue zhu Jing Zhai yuan Xu yan hong Jian-lin Hu yun-Peng Peng Zhen-Bo Zhang yu Wu Zheng Li Department of ART Department of AndrologyCenter for Men’s HealthInstitute of Urologic Medical CenterShanghai General HospitalShanghai Key Lab of Reproductive MedicineShanghai Jiao Tong UniversityShanghai 200080China Clinical Research Center for Reproduction and Genetics in Hunan Province Reproductive and Genetic Hospital of CITIC-XiangyaChangsha 410000China Department of Urology Affiliated Hospital of Xuzhou Medical UniversityXuzhou 221000China

Cryopreservation of rare testicular-retrieved spermatozoa for intracytoplasmic sperm injection(ICSI)in patients with severe oligozoospermia and azoospermia remains a major challenge in clinical *** study evaluated the Cryopiece system as a potential technique to cryopreserve rare human spermatozoa for *** numbers of ejaculated(24 patients)and testicular(13 patients)spermatozoa were cryopreserved using the Cryopiece *** total number of recovered spermatozoa and motility were assessed after ***-seven couples underwent ICSI using spermatozoa cryopreserved by the Cryopiece system,and ICSI outcomes(rates of fertilization,embryo cleavage,and clinical pregnancy)were *** average sperm post-thaw retrieval rate was 79.1%,and motility was 29.7%.Ejaculated spermatozoa had a higher post-thaw motility(32.5%)than testicular spermatozoa(21.8%;P=0.005).ICSI achieved a fertilization rate of 61.9%,embryo cleavage rate of 84.6%,and clinical pregnancy rate of 43.3%.The ICSI outcomes in the ejaculated and testicular frozen-thawed spermatozoa were *** oocyte activation(AOA)after ICSI with motile(72.1%)or immotile(71.9%)spermatozoa resulted in a significantly higher fertilization rate than that when using motile spermatozoa without AOA(52.0%;P=0.005).However,AOA did not enhance the clinical pregnancy rate(55.6%or 40.0%vs 35.3%;P=0.703).The Cryopiece system is simple and useful for the cryopreservation of small numbers of ejaculated or testicular spermatozoa for ICSI in patients with severe oligozoospermia or nonobstructive azoospermia.

关键词： Cryopiece system intracytoplasmic sperm injection outcome rare sperm cryopreservation