骨髓移植修复放疗损伤小鼠卵巢的实验研究
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摘要
随着对肿瘤放疗领域研究的不断深入,肿瘤患者预后明显改善,患者生存率明显提高。据美国1995年～2005年间数据统计,所有诊断浸润癌患者(无论年龄大小)5年生存率接近60%。其中年轻女性肿瘤患者5年生存率远高于60%,治疗所致的医源性损害已逐渐得到重视。如放疗常见的一个并发症卵巢毒性引起的早绝经和不育严重影响患者的生活质量和自信。近年有关放疗对卵巢损害的机制以及预防卵巢受损的研究受到了肿瘤学、生殖医学、放射防护学等学科的关注,本文就利用骨髓细胞的发育潜能来修复放疗损伤卵巢组织的可能性及其机制进行了初步实验研究。
本研究的目的是通过观察骨髓移植对放疗引起的卵巢受损小鼠的修复效果,比较同性/异性骨髓移植对放射引起的卵巢损伤修复的差异,以及推测其损伤修复机制。观察的指标包括:血清中FSH、E_2含量,卵巢系数(OI),卵巢组织的组织形态及是否出现动情周期。
本论文分为三个部分。第一部分为研究骨髓移植是否对放疗引起的小鼠卵巢损伤有恢复作用;第二部分为比较同性或异性骨髓移植对放疗引起的小鼠卵巢损伤的修复是否有差异;第三部分为关于放疗损伤卵巢组织功能恢复机制的研究。各部分的实验方法如下:
1.骨髓移植对放疗引起的小鼠卵巢损伤修复作用的实验中,我们采用140只SPF级8～10周龄雌性昆明小鼠,除20只作为骨髓移植供体,其余120只随机分为平均三组:正常对照组,单次放疗组,骨髓移植组。正常对照组,不采取任何措施;单次放疗组,单次放疗后不采取任何措施;骨髓移植组,单次放疗后24h进行骨髓移植。骨髓移植后于第1,5,15,30天四个时间点分别从各组中取10只小鼠检测血清中E_2,FSH水平并测定OI,取卵巢组织做病理切变观察;采用析因方差方法进行统计。
2.同性或异性骨髓移植对放疗引起的小鼠卵巢损伤的修复实验中,我们采用175只SPF级8～10周龄雌性昆明小鼠,15只SPF级8～10周龄雄性昆明小鼠。其中15只雄性小鼠作为雄性骨髓移植供体,15只雌性小鼠作为雌性骨髓移植供体。余下160只雌性小鼠随机分为四组:正常对照组,单次放疗组,同性骨髓移植组,异性骨髓移植组。正常对照组,不采取任何措施;单次放疗组,单次放疗后不采取任何措施;异性骨髓移植组,单次放疗后24h内移植雄性小鼠骨髓细胞;同性骨髓移植组,单次放疗后24h内移植雌性小鼠骨髓细胞。于骨髓移植后第1,5,15,30天每个时间点分别从各组中取10只小鼠检测血清中E_2,FSH水平,并测定OI,并取卵巢组织做病理切变观察;采用析因方差方法进行统计。
3.骨髓细胞移植到放疗后小鼠的体内实验中,我们采用了22只雌性ICR小鼠,其中10只作为荧光骨髓移植受体,6只作为单次放疗组,6只作为空白对照组。另有5只带有绿色荧光标记的转基因ICR小鼠作为骨髓移植供体。单次放疗组单次放疗后不采取任何措施;空白对照组,不采取任何措施;骨髓移植组,单次放疗后24h移植荧光小鼠骨髓细胞。骨髓移植后的第15,30天分别取对照组3只,单次放疗组3只,荧光骨髓移植组5只小鼠的卵巢组织做冰冻切变观察是否出现带有荧光标记的卵细胞。
实验结果如下:
1.放射后放疗后骨髓移植组(E_2:10.83ng/L,FSH:2.94IU/L,OI:2.16),单次放疗组(E_210.30ng/L,FSH2.84IU/L,OI:2.24)的激素水平、卵巢系数与正常组(E_210.83ng/L,FSH2.42IU/L,OI:2.70)差异显著,在放疗后第15、30天,单次放疗组成熟卵泡数量减少,卵巢血管增生,有纤维化;骨髓移植组可见原始卵泡,初级卵泡,次级卵泡,成熟卵泡,血管较丰富。血清中E_2、FSH水平及OI在放疗后第15、30天骨髓移植组(E_223.02ng/L,FSH2.59IU/L,OI:2.60;E_223.92ng/L,FSH2.24IU/L,OI:2.76)与单次放疗组(E_218.17ng/L,FSH2.98IU/L,OI:2.34;E_219.62ng/L,FSH3.05IU/L,OI:2.47)有显著性差异。(血清中E_2、FSH水平及卵巢系数在第15天时的F值分别为15.676,2.854,27.215;P值分别为0.000,0.000,0.045;第30天时的F值分别为12.610,15.657,19.709;P值分别为0.000,0.000,0.000)。
2.放疗后初期同性骨髓移植组、异性骨髓移植组、单次放疗组的E_2,FSH,OI分别为(12.12ng/L,3.59IU/L,2.23;12.10ng/L,3.56IU/L,2.27;10.38ng/L,3.29IU/L,2.22),与正常对照组(25.35ng/L,2.36IU/L,2.83)差异显著;在骨髓移植第15、30天,单次放疗组、异性骨髓移植组成熟卵泡数量持续减少,卵巢血管增生,有纤维化,较实验初期未见明显好转;同性骨髓移植组可见原始卵泡,初级卵泡,次级卵泡,成熟卵泡,血管较丰富。在第15天同性骨髓移植组(E_218.38ng/L,FSH3.10IU/L,OI:2.59)血清中E_2、FSH水平及OI与单次放疗组(E_210.83ng/L,FSH2.42IU/L,OI:2.70)、异性移植组(E_212.54ng/L,FSH3.88IU/L,OI:2.46)有显著性差异,与正常对照组(E_225.39ng/L,FSH2.39IU/L,OI:2.82)接近,并可以继续好转至30日。(血清中E_2、FSH水平及OI在第15天时的F值分别为124.038、95.431、15.346;P值分别为0.000、0.000、0.000;在第30天时的F值分别为180.967、190.495、10.968;P值分别为0.000、0.000、0.000)
3.骨髓移植后第15,30天,冰冻切片观察各组卵巢内卵细胞生长情况时发现,移植荧光小鼠骨髓的雌性小鼠卵巢中均发现不同程度的荧光细胞,而对照组别未发现荧光细胞。
通过以上研究,我们可以判断出骨髓移植对由放疗引起的小鼠卵巢损伤具有一定恢复作用,但是只有同性之间的骨髓移植才对卵巢恢复起作用,同时证明了外源性的骨髓干细胞不仅可以促进小鼠本身卵巢功能的恢复,而且可以在放疗后小鼠卵巢中继续发育成为卵细胞。
With rapid development in the field of tumor radiotherapy,the fidelity of prognosis and surviving rate of patients with cancer improved obviously.According to the statistical results from 1995～2005 in U.S.A,the survive rate of the infiltrating carcinoma patients reached as high as 60%.With the improvement of long term survive rate of young female patients,more and more attention was paid to iatrogenic harm.For example,long term physiological and psychological hurt.One of long term complication in radiotherapy is ovary toxicity.This is due to the finity of ovary germ cells,which can't be regenerated.Therefore,to many young female patients who expect normal pregnant,the sterility and early menopause caused by radiotherapy affect their life quality and self-confidence seriously.Recently,the study on the mechanisms and prevention of ovary toxicity caused by radiotherapy aroused the interest of researchers in the field of phymatology,reproductive medicine,radiative prevention.This paper initially studied the feasibility and mechanisms of the recovery of radiotherapic ovary damage by marrow transplantation.
The aim of this paper is to determine the recovery level of radiotherapic ovaries after marrow transplantation by observing pathological morphous,testing FSH,E_2 level in blood serum of mouse and checking whether estrous cycle was reoccurred. Then,after observing the differences between homogeneity marrow transplantation group(HOMT) and heterosexual(HEMT) marrow transplantation groupto,a new mechanism for ovary recovery can be provided.
The study was divided into three parts.The first part mainly studied the fuction of ovary recovery by transplanting marrow into radiotherapic mouse;the second part discussed different ovary recovery levels between HOMT and HEMT;the third part initially explained the possible mechanism of recovery in this way.
1.In the first experiment,we used 140 8～10 weeks(w) Kunming female mouse(SPF,same in the following experiments),20 as transplant donor,40 as control group which didn't receive any treatment;40 as radiotherapy group which were radiated once;40 as transplant group which were performed marrow transplant within 24h after radiation.On the day 1~(st),5~(th),15~(th),30~(th) we selected 10 mouse from each groups respectively to detected the E_2,FSH in blood serum,weighted their ovary index and made pathological section of ovary.
2.In the second experiment,we used 175 SPF 8～10w Kunming female mouse and 15 SPF 8～10w male Kunming mouse.15 male mouse as transplant donor and 15 female mouse as transplant donor,40 as control group which didn't adopt any treatment;40 as radiotherapy group which radiated once only;40 as transplant group which performed female marrow transplant within 24h after radiation,40 as transplant group which performed male marrow transplant within 24h after radiation. On the day 1~(st),5~(th),15~(th),30~(th) we selected 10 mouse from each groups respectively to detected the E_2,FSH in blood serum and weighted ovary index and made pathological section of ovary.
3.In the third experiment,we used 22 female ICR mouse,5 female ICR fluorescent mouse.6 ICR mouse as radiotherapy group which radiated only once,6 ICR mouse as control group which didn't have any treatment,5 fluoresent mouse as marrow transplant donor,10 ICR as marrow transplant group which transplant fluorescent marrow within 24h after radiation.On day 15~(th),30~(th),we select half of the group perpform ovary frozen section respectively.
In the experiment on whether marrow transplant is good for ovary recovery harmed by radiotherapy,on day 15~(th) and 30~(th),we observed marrow transplant group reappeared more primordial follicle,primary follicle,secondary follicle and mature follicle compared with radiotherapy groupand,and the vessels in ovary was comparatively abundant.The number of ovum in radio therapy group was dropped dramatically.The E_2 and FSH level in blood serum and ovary index between radiotherapy group and marrow transplant group has significant differences on day 15~(th) and 30~(th).(The F value of E_2,FSH,ovary index on 15~(th) day was 15.676,2.854, 27.215 respectively,the P value of E_2,FSH,ovary index was 0.000,0.000,0.045 respectively;The F value of E_2,FSH,ovary index on 30~(th) day was 12.610,15.657, 19.709 respectively,the P value of E_2,FSH,ovary index was 0.000,0.000,0.000).
In the experiment on whether marrow from different sex is different for ovary recovery harmed by radiotherapy,on day 15~(th) and 30~(th),compared with radiotherapy group,we observed female marrow transplant group reappeared primordial follicle, primary follicle,secondary follicle and mature follicle,the vessels in ovary was comparatively was abundant.The number of ovum in radiotherapy group and male marrow transplant group was dropped dramatically.The E_2 and FSH level in blood serum and OI between radiotherapy group,HEMT and HOMT had significant differences on day 15~(th) and 30~(th).(The F value of E_2,FSH,OI on 15~(th) day was 124.038,95.431,15.346 respectively,the P value of E_2,FSH,ovary index was0.000, 0.000,0.000 respectively;The F value of E_2,FSH,OI on 30~(th) day was 180967,190.495,10.968 respectively,the P value of E_2,FSH,ovary index was0.000, 0.000,0.000)
In the third part,we found GFP cells in the GFP marrow transplantation group on d15 and d30,while in other groups we didn't found any GFP cells.
According to the results above,a conclustion can be drawed that marrow transplantion of same sex contributes to the recovery of ovary harmed by radiation. The marrow cells from extrinsic sourc have effect on both ovary itself and can be further developed into ovums.
本研究的目的是通过观察骨髓移植对放疗引起的卵巢受损小鼠的修复效果,比较同性/异性骨髓移植对放射引起的卵巢损伤修复的差异,以及推测其损伤修复机制。观察的指标包括:血清中FSH、E_2含量,卵巢系数(OI),卵巢组织的组织形态及是否出现动情周期。
本论文分为三个部分。第一部分为研究骨髓移植是否对放疗引起的小鼠卵巢损伤有恢复作用;第二部分为比较同性或异性骨髓移植对放疗引起的小鼠卵巢损伤的修复是否有差异;第三部分为关于放疗损伤卵巢组织功能恢复机制的研究。各部分的实验方法如下:
1.骨髓移植对放疗引起的小鼠卵巢损伤修复作用的实验中,我们采用140只SPF级8～10周龄雌性昆明小鼠,除20只作为骨髓移植供体,其余120只随机分为平均三组:正常对照组,单次放疗组,骨髓移植组。正常对照组,不采取任何措施;单次放疗组,单次放疗后不采取任何措施;骨髓移植组,单次放疗后24h进行骨髓移植。骨髓移植后于第1,5,15,30天四个时间点分别从各组中取10只小鼠检测血清中E_2,FSH水平并测定OI,取卵巢组织做病理切变观察;采用析因方差方法进行统计。
2.同性或异性骨髓移植对放疗引起的小鼠卵巢损伤的修复实验中,我们采用175只SPF级8～10周龄雌性昆明小鼠,15只SPF级8～10周龄雄性昆明小鼠。其中15只雄性小鼠作为雄性骨髓移植供体,15只雌性小鼠作为雌性骨髓移植供体。余下160只雌性小鼠随机分为四组:正常对照组,单次放疗组,同性骨髓移植组,异性骨髓移植组。正常对照组,不采取任何措施;单次放疗组,单次放疗后不采取任何措施;异性骨髓移植组,单次放疗后24h内移植雄性小鼠骨髓细胞;同性骨髓移植组,单次放疗后24h内移植雌性小鼠骨髓细胞。于骨髓移植后第1,5,15,30天每个时间点分别从各组中取10只小鼠检测血清中E_2,FSH水平,并测定OI,并取卵巢组织做病理切变观察;采用析因方差方法进行统计。
3.骨髓细胞移植到放疗后小鼠的体内实验中,我们采用了22只雌性ICR小鼠,其中10只作为荧光骨髓移植受体,6只作为单次放疗组,6只作为空白对照组。另有5只带有绿色荧光标记的转基因ICR小鼠作为骨髓移植供体。单次放疗组单次放疗后不采取任何措施;空白对照组,不采取任何措施;骨髓移植组,单次放疗后24h移植荧光小鼠骨髓细胞。骨髓移植后的第15,30天分别取对照组3只,单次放疗组3只,荧光骨髓移植组5只小鼠的卵巢组织做冰冻切变观察是否出现带有荧光标记的卵细胞。
实验结果如下:
1.放射后放疗后骨髓移植组(E_2:10.83ng/L,FSH:2.94IU/L,OI:2.16),单次放疗组(E_210.30ng/L,FSH2.84IU/L,OI:2.24)的激素水平、卵巢系数与正常组(E_210.83ng/L,FSH2.42IU/L,OI:2.70)差异显著,在放疗后第15、30天,单次放疗组成熟卵泡数量减少,卵巢血管增生,有纤维化;骨髓移植组可见原始卵泡,初级卵泡,次级卵泡,成熟卵泡,血管较丰富。血清中E_2、FSH水平及OI在放疗后第15、30天骨髓移植组(E_223.02ng/L,FSH2.59IU/L,OI:2.60;E_223.92ng/L,FSH2.24IU/L,OI:2.76)与单次放疗组(E_218.17ng/L,FSH2.98IU/L,OI:2.34;E_219.62ng/L,FSH3.05IU/L,OI:2.47)有显著性差异。(血清中E_2、FSH水平及卵巢系数在第15天时的F值分别为15.676,2.854,27.215;P值分别为0.000,0.000,0.045;第30天时的F值分别为12.610,15.657,19.709;P值分别为0.000,0.000,0.000)。
2.放疗后初期同性骨髓移植组、异性骨髓移植组、单次放疗组的E_2,FSH,OI分别为(12.12ng/L,3.59IU/L,2.23;12.10ng/L,3.56IU/L,2.27;10.38ng/L,3.29IU/L,2.22),与正常对照组(25.35ng/L,2.36IU/L,2.83)差异显著;在骨髓移植第15、30天,单次放疗组、异性骨髓移植组成熟卵泡数量持续减少,卵巢血管增生,有纤维化,较实验初期未见明显好转;同性骨髓移植组可见原始卵泡,初级卵泡,次级卵泡,成熟卵泡,血管较丰富。在第15天同性骨髓移植组(E_218.38ng/L,FSH3.10IU/L,OI:2.59)血清中E_2、FSH水平及OI与单次放疗组(E_210.83ng/L,FSH2.42IU/L,OI:2.70)、异性移植组(E_212.54ng/L,FSH3.88IU/L,OI:2.46)有显著性差异,与正常对照组(E_225.39ng/L,FSH2.39IU/L,OI:2.82)接近,并可以继续好转至30日。(血清中E_2、FSH水平及OI在第15天时的F值分别为124.038、95.431、15.346;P值分别为0.000、0.000、0.000;在第30天时的F值分别为180.967、190.495、10.968;P值分别为0.000、0.000、0.000)
3.骨髓移植后第15,30天,冰冻切片观察各组卵巢内卵细胞生长情况时发现,移植荧光小鼠骨髓的雌性小鼠卵巢中均发现不同程度的荧光细胞,而对照组别未发现荧光细胞。
通过以上研究,我们可以判断出骨髓移植对由放疗引起的小鼠卵巢损伤具有一定恢复作用,但是只有同性之间的骨髓移植才对卵巢恢复起作用,同时证明了外源性的骨髓干细胞不仅可以促进小鼠本身卵巢功能的恢复,而且可以在放疗后小鼠卵巢中继续发育成为卵细胞。
With rapid development in the field of tumor radiotherapy,the fidelity of prognosis and surviving rate of patients with cancer improved obviously.According to the statistical results from 1995～2005 in U.S.A,the survive rate of the infiltrating carcinoma patients reached as high as 60%.With the improvement of long term survive rate of young female patients,more and more attention was paid to iatrogenic harm.For example,long term physiological and psychological hurt.One of long term complication in radiotherapy is ovary toxicity.This is due to the finity of ovary germ cells,which can't be regenerated.Therefore,to many young female patients who expect normal pregnant,the sterility and early menopause caused by radiotherapy affect their life quality and self-confidence seriously.Recently,the study on the mechanisms and prevention of ovary toxicity caused by radiotherapy aroused the interest of researchers in the field of phymatology,reproductive medicine,radiative prevention.This paper initially studied the feasibility and mechanisms of the recovery of radiotherapic ovary damage by marrow transplantation.
The aim of this paper is to determine the recovery level of radiotherapic ovaries after marrow transplantation by observing pathological morphous,testing FSH,E_2 level in blood serum of mouse and checking whether estrous cycle was reoccurred. Then,after observing the differences between homogeneity marrow transplantation group(HOMT) and heterosexual(HEMT) marrow transplantation groupto,a new mechanism for ovary recovery can be provided.
The study was divided into three parts.The first part mainly studied the fuction of ovary recovery by transplanting marrow into radiotherapic mouse;the second part discussed different ovary recovery levels between HOMT and HEMT;the third part initially explained the possible mechanism of recovery in this way.
1.In the first experiment,we used 140 8～10 weeks(w) Kunming female mouse(SPF,same in the following experiments),20 as transplant donor,40 as control group which didn't receive any treatment;40 as radiotherapy group which were radiated once;40 as transplant group which were performed marrow transplant within 24h after radiation.On the day 1~(st),5~(th),15~(th),30~(th) we selected 10 mouse from each groups respectively to detected the E_2,FSH in blood serum,weighted their ovary index and made pathological section of ovary.
2.In the second experiment,we used 175 SPF 8～10w Kunming female mouse and 15 SPF 8～10w male Kunming mouse.15 male mouse as transplant donor and 15 female mouse as transplant donor,40 as control group which didn't adopt any treatment;40 as radiotherapy group which radiated once only;40 as transplant group which performed female marrow transplant within 24h after radiation,40 as transplant group which performed male marrow transplant within 24h after radiation. On the day 1~(st),5~(th),15~(th),30~(th) we selected 10 mouse from each groups respectively to detected the E_2,FSH in blood serum and weighted ovary index and made pathological section of ovary.
3.In the third experiment,we used 22 female ICR mouse,5 female ICR fluorescent mouse.6 ICR mouse as radiotherapy group which radiated only once,6 ICR mouse as control group which didn't have any treatment,5 fluoresent mouse as marrow transplant donor,10 ICR as marrow transplant group which transplant fluorescent marrow within 24h after radiation.On day 15~(th),30~(th),we select half of the group perpform ovary frozen section respectively.
In the experiment on whether marrow transplant is good for ovary recovery harmed by radiotherapy,on day 15~(th) and 30~(th),we observed marrow transplant group reappeared more primordial follicle,primary follicle,secondary follicle and mature follicle compared with radiotherapy groupand,and the vessels in ovary was comparatively abundant.The number of ovum in radio therapy group was dropped dramatically.The E_2 and FSH level in blood serum and ovary index between radiotherapy group and marrow transplant group has significant differences on day 15~(th) and 30~(th).(The F value of E_2,FSH,ovary index on 15~(th) day was 15.676,2.854, 27.215 respectively,the P value of E_2,FSH,ovary index was 0.000,0.000,0.045 respectively;The F value of E_2,FSH,ovary index on 30~(th) day was 12.610,15.657, 19.709 respectively,the P value of E_2,FSH,ovary index was 0.000,0.000,0.000).
In the experiment on whether marrow from different sex is different for ovary recovery harmed by radiotherapy,on day 15~(th) and 30~(th),compared with radiotherapy group,we observed female marrow transplant group reappeared primordial follicle, primary follicle,secondary follicle and mature follicle,the vessels in ovary was comparatively was abundant.The number of ovum in radiotherapy group and male marrow transplant group was dropped dramatically.The E_2 and FSH level in blood serum and OI between radiotherapy group,HEMT and HOMT had significant differences on day 15~(th) and 30~(th).(The F value of E_2,FSH,OI on 15~(th) day was 124.038,95.431,15.346 respectively,the P value of E_2,FSH,ovary index was0.000, 0.000,0.000 respectively;The F value of E_2,FSH,OI on 30~(th) day was 180967,190.495,10.968 respectively,the P value of E_2,FSH,ovary index was0.000, 0.000,0.000)
In the third part,we found GFP cells in the GFP marrow transplantation group on d15 and d30,while in other groups we didn't found any GFP cells.
According to the results above,a conclustion can be drawed that marrow transplantion of same sex contributes to the recovery of ovary harmed by radiation. The marrow cells from extrinsic sourc have effect on both ovary itself and can be further developed into ovums.
引文
[1]姚琴,李彦。放疗的副作用及其护理[J],国外医学护理学分,2001年第20卷第3期:134-136
[2]Tilly JL.Kolesnick RN.Sphingolipids apoptosis cancer treatments and the ovary:investigating a crime against female fertility[J].Biochem Biophys Acta.2002,30(23):135-138
[3]Morice P,Juncker I,Rey A,et al.Ovarian transposition for patients with cervical carcinoma treated by radiosurgical combination[J].Fertil Steril,2000,74:743-748
[4]Revel A,Laufer N.Protecting female fertility from cancer therapy[J].Mol Cell Endocrinol,2002,187:83-91
[5]Kim SS,Battaglia DE,Soules MR.The future of human ovarian cryopreservation and transplantation:fertility and beyond[J].Fertil Steril,2001,75:1049-56
[6]Trew GH,Filippis DS,et al.Optimizing gonadotrophin-releasing hormone antagonist protocol[J].Hum Fertil,2002,5(1):41-48
[7]徐安然 邓晓惠 放化疗致卵巢损害的机制及其组织学改变[J],国外医学计划生育/生殖健康分册,2006年25卷第6期:333-336
[8]Lutchman SK,Davies M,Chatterjee R.Fertility in female cancer survivor:pathophysiology,preservation and the role of ovarian reserve testing[J].Hum Reprod Update,2005,11(1):69-89
[9]Wallace WH,Thomson AB,Kelsey TW.The radiosensitivity of the human oocyte[J].Hum Reprod.2003,18(1):117-121
[10]Samuel K,Buyuk E Fertility preservation in female cancer patients:current developments and future directions[J].Fertil Steril,2006,85(1):1-11
[11].Feniche IP.Medical preservation of ovarian function before and during radiochemotherapy[J].Gynecol Obstet Fertil,2005,33(9):610-614。
[12]王皓,邹正辉,汪涛。云芝糖肽对受照小鼠骨髓移植的影响[J],Acta acadiciac medicine 1997,17(5):828-830
[13]刘怡,王松灵。干细胞的研究进展[J],北京口腔医学,2006,14(4):289-294
[14]Blau HM,Brazelton TR,Weimann JM.The evolving concept of a stem cell entity of function[J].Cell,2001,105(7):829-841
[15]丁强,贾心善。成体干细胞分化调控研究的新进展[J],日本医学介绍,2004,25(1):39-40
[16]Lakshmipathy U,Verfaillic C.Stem cell plasticity[J].Blood Rev,2005,19(1):29-38
[17]黄海霞,汤学明。成体干细胞多能性研究进展[J],生命科学,2002,14(3):129-135
[18]李建生,刘敬霞。成体干细胞在中枢神经系统疾病修复的效应[J],中国组织工程研究与临床康复2007,Ⅱ(11):2144-2149
[19]Ferrari G,Cusella G,Angelis D,et al.Muscle regeneration by bone marrow derived myogenic progenitors[J].Science,1998,279:1528-30
[20]Theise ND,Nimmakayaly M,Gardner R,et al.Liver from bone marrow stem cells can differentiate into hepatocytes in vivo[J].Nat Med,2000,32(1):11-16
[21]Orlic D,Kajstura J,Chimenti S,et al.Bone marrow cells regenerate infarcted myocarduim[J].Nature,2001,410:701-705
[22]Luo LL.Infertility and sterility[M].Beijing:People Healthy Press.1998:5-6
[23]Smikel CB,Dandekar PV,Schrioch ED,et al.Elevated ovarian follicular fluid stem cell factor concentrations are associated with improved pregnancy rates in vitro fertilization cycles[J].Fertil Steril,1998,69(1):70-72
[24]Smikel CB,Dandekar PV,Schrioch ED,et al.Elevated ovarian follicular fluid stem cell factor concentrations are associated with improved pregnancy rates in vitro fertilization cycles[J].Fertil Steril,1998,69(1):70-72
[25]Tanikawa M,Harada T,Zwabe T,Presence of stem cell factor in follicular fluild and its expression in the human ovary[J].Fertil Steril,2000,73(6):1259-60
[26]Parrott JA,Skinner MK,Kit-ligand and stem cell factor induces primordial follicle development and initiates folliculogenesis[J].Endocrinology 1999,140(9):4262-71
[27]Parrott JA,Skinner MK,kit-ligand action on ovarian stromal cells:effects on the cell recruitment and steroid production[J].Mol Reprod Dev 2000,55(1):55-64
[28]Castrillon DH,Miao L,Kollipara R,et al.Suppression of ovarian follicle activation in mice by the transcription factor FOXO3a[J].Science 2003,301(5630):215-218
[29]Reddy P,Shen L,Ren C,et al.Activation of Akt and suppression of FKHRL1 in mouse and rat oocyte by stem cell factor during follicular activation and development[J].Dev Biol,2005,281(2):160-170
[30]余荣娇,徐斯凡。干细胞因子在精子发生中的作用[J],江西医学院学报2004,44(4):114-116
[31]陈振文,白照岱。胚胎干细胞向生殖细胞的分化研究进展[J],中国男科学杂志2007,21(2):1-5
[32]Young LE.Focus on stem cells in reproduction[J]. Reproduction,2006,132(5):671-672
[33]Zuckerman S,The number of oocytes in the mature ovary[J].Recent Prog.Hom.Res.6:63-108
[34]杨晓菁,姚红军,王玉凤。哺乳动物卵母细胞凋亡的研究进展[J],动物学杂志 2004,39(6):101-106
[35]Hirshfield AN.Development of follicles in the mammalian ovary[J].Int Rev Cytol.1991,124:43-101
[36]罗丽莉,黄菊,傅玉才。内分泌因子,生长因子、细胞因子与卵巢卵泡的发育[J],中国组织工程研究与临床康复 2007,11(19):3821-30。
[37]Joshua Johnson,Jacqueline Canning,Tomoko Kaneko,Germline stem cells and follicular renewal in the postnatal mammalian ovary[J],Nature,2004(428):145-153。
[38]Roger G.Gosden Germline stem cells in the postnatal ovary:is the ovary more like a testis?[J]Human reproduction update,2004(10) 193-195
[39]Bukovsky A,Svetlikova M,Caudle MR.Oogenesis in cultures derived from adult human ovaries[J].Reprod Biol Endocrinol,2005,3(1):17-30
[40]Hubner K,Fumann G,Christenson LK,et al.Derivation of oocytes from mouse embrynic stem cells[J].Science 2003,300(23):1251-56
[41]Jonoathan L,Tilly Commuting the death sentence:how oocytes strive to survive[J],nature,2001(2):838-840
[42]Joshua Johnson,Jessamyn Bagley,Malgorzata Skaznik-Wikiel Oocyte generation in adult mammalian ovaries by putative germ cells in bone marrow and peripheral blood[J],Cell,Vol122:303-315
[43]侯友贤。恶性肿瘤放疗与康复[M],人民军医出版社,2005年06月:20-29
[44]于金明。2005年肿瘤精确放疗进展(修订版)[M],第三届济南国际肿瘤放射治疗新进展暨第二届全国中青年放射肿瘤学术研讨会论文汇编:31-36
[45]王奇璐。肿瘤化疗、放疗201个怎么办(第二版)[M],中国协和医科大学出版社,2004,08:9-11
[46]姚琴,李彦.放疗的副作用及护理[J],国外医学护理学分册,2001,20(3):134-137
[47]孙建衡。妇科恶性肿瘤放疗的几个问题[J],肿瘤学杂志,2006,12(5):357-360
[48]Seymour JF.Ovarian tissue cryoreservation for cancer patients:who is appropreiate?[J].Reprod Fertil,2001(13)81-89
[49]Blumenfeld Z.Preservation of fertily and ovarian fuction and minimalization of chemotherapy associated gonadotoxity and premature ovarian failure:the role of inhibin-A and-B as marks[J].Mol Cell Endorinol,2002,187:93-105
[50]Richardson SJ,Senikas V,Nelson JF.Follicular depletion during the menoarusal transition:evidence for accelerated loss and ultimate exhaustion[J].J.Clin.Endocrinal.Metab.1997(65):1231-37
[51]Feniche IP.Medical preservation of ovarian function before and during radiochemotherapy[J].Gynecol Obstet Fertil,2005,33(9):610-614
[52]Louise D,Bath LE,Anderson RA,et al.Hypothalarmic pituitary ovarian dysfuction after radiotherapic prepubertal ad cranial irradiation for acute leukaemia[J].Hum Reprod,2001,16(9):1838-44
[53]Meirow D,Nugent D.The effects of radiotherapy and chemotherapy on female reproduction[J].Hum Reprod Update,2001,7(6):535-543
[54]Thibaud E,Rodriguez Macias K,Irivin C,et al.Ovarian functino after bone marrow transplantation during childhood[J].Bone Marrow Transplant,1998,21:287-290
[55]Chiarelli AM,Marrett LD,Darlington G,et al.Early menopause and infertility in female after treatment for chindhood cancer diagnosed in 1964-1988 in Ontario,Canada[J].Am JE Epidemiol,1999,150:245-254
[56]Gosden RG,Wade JC,Fraser HM,et al.Impact of congenital hypogonadotropism on the radiation sensitivity of the mouse ovary[J].Hum Reprod,1997,12:2483-2488
[57]Bertram JS.The molecular biology of cancer[J].Mol Aspects Med,2000,21(6):167-233
[58]吴郁,向阳。化疗及放疗对卵巢的影响及预防[J],国外医学妇产科分册,2003,30(3):166-170
[59]Almodin CG,Minguett VC,Meister H,et al.Recovery of natural fertility after grafting of ciyopreserved germinative tissue ewes subjected to radiotherapy[J].Fertil Steril,2004,81(1):160-164
[60]Blumenenfeld Z,Shapiro D,Shteinberg M,et al.Preservation of fertility and ovarian function and minimizing gonado-toxicity in young women with systemic lupus erythematosus treated bu radiotherapy[J].Lupus,2000,9(6):401-405
[61]Pereyra-Pacheco B,Mendez Ribas.Use of GnRH analogs for functional protection of the ovary and preservation of fertility during cancer treatment in adolescent a preliminary report[J],Gynecol Oncol,2001,81(3):391-397
[62]Bath LE,Wallace WH,Shaw MP,et al.Depletion of ovarian reserve in young women after treatment for cancer in childhood:detection by anti-mullertian hormone,inhibin B and ovarian ultrasound[J],Hum Reprod,2003,18(11):2368-74
[63]Domans MM,Demylle D,Martine B,et al.Efficacy of in vitro fertilization after radiotherapy[J],Fertil Steril,2005,83(4):897-901
[64]Berek JS,Hacker NF.Practical Gynecologic oncology[J],Lippincott Willians Wilkins,2004,23(4):119-162
[65]Oktay K,Buyuk E,Veeck L,et al.Embryo development after heterotopic transplantation of cryopreserved ovarian tissue[J],Lancet,2004,363(9412:837-840)
[66]Meirow D.Ovarian injury and mordern options to preserve fertility in female concer patient treated with hign dose radio-chemotherapy for hemato-oncological meoplasias and other cancers[J].Leukema Lymphoma,1999,33(12).65-76
[67]Morice P,Juncker L,Ray A,et al.Ovarian transposition for patients with cervical carcinoma treated by radiosurgical combination[J],Fertil Steril,2000,74:743-748
[68]Blumenfeld Z,Dann E,Avivi I,et al.Fertility after treatment for Hodgkin's disease[J].Ann Oncol,2002,13[suppl 11]:138-147
[69]Morita Y,Perez GI,Paris EOocyte apoptosis is suppressed by disruption of the acid sphingomylinase gene or by sphingosine-1-phosphate therapy[J],Nat Med,2000,6:1109-1114
[70]潘兴华,段连宁,韩毅冰。成体干细胞的可塑性及临床意义[J],Journal of tropical Medicine,2001,1(2):158-163
[71]张静。成体干细胞的应用[J],生物学杂志,2005,22(4):64。
[72]Theise ND,Nimmakayalu M,Gardner T,et al.Liver from bone marrow in human[J].Hepatology,2000,32(11):11-16
[73]刘芊,高敏。骨髓间充质干细胞向肝细胞分化的研究[J].国际消化杂志,2007,27(1):7-10
[74]Clarke DL,Johansson CB,Frisen J,et al.Generalized potential of adult stem cells[J].Science,2002,286:1660-1661
[75]Pittenger MF,Mackay AM,Beck SC,et al.Multilineage potential of adult human mesenchymal stem cells[J],Science,1999,284:143-147
[76]Filip S,Mokr J,Hruska I.Adult stem cells and their importance in cell therapy[J].Folia Biologica,2003,49:9-14
[77]David AP.Current science of regenerative medicine with stem cells[J].Journal of Investive Medicine.2006,54(1):33-35
[78]Vieyra DS,Jackson KA,Goodell MA.Plasticity and tissue regenerative potential of bone marrow-derived cells[J].Stem Cell Rev.2005;1(1):65-69。
[79]潘兴华,张步振。自体骨髓干细胞的临床应用及存在的问题[J],中国医疗前沿,2007,30(15):25-28
[2]Tilly JL.Kolesnick RN.Sphingolipids apoptosis cancer treatments and the ovary:investigating a crime against female fertility[J].Biochem Biophys Acta.2002,30(23):135-138
[3]Morice P,Juncker I,Rey A,et al.Ovarian transposition for patients with cervical carcinoma treated by radiosurgical combination[J].Fertil Steril,2000,74:743-748
[4]Revel A,Laufer N.Protecting female fertility from cancer therapy[J].Mol Cell Endocrinol,2002,187:83-91
[5]Kim SS,Battaglia DE,Soules MR.The future of human ovarian cryopreservation and transplantation:fertility and beyond[J].Fertil Steril,2001,75:1049-56
[6]Trew GH,Filippis DS,et al.Optimizing gonadotrophin-releasing hormone antagonist protocol[J].Hum Fertil,2002,5(1):41-48
[7]徐安然 邓晓惠 放化疗致卵巢损害的机制及其组织学改变[J],国外医学计划生育/生殖健康分册,2006年25卷第6期:333-336
[8]Lutchman SK,Davies M,Chatterjee R.Fertility in female cancer survivor:pathophysiology,preservation and the role of ovarian reserve testing[J].Hum Reprod Update,2005,11(1):69-89
[9]Wallace WH,Thomson AB,Kelsey TW.The radiosensitivity of the human oocyte[J].Hum Reprod.2003,18(1):117-121
[10]Samuel K,Buyuk E Fertility preservation in female cancer patients:current developments and future directions[J].Fertil Steril,2006,85(1):1-11
[11].Feniche IP.Medical preservation of ovarian function before and during radiochemotherapy[J].Gynecol Obstet Fertil,2005,33(9):610-614。
[12]王皓,邹正辉,汪涛。云芝糖肽对受照小鼠骨髓移植的影响[J],Acta acadiciac medicine 1997,17(5):828-830
[13]刘怡,王松灵。干细胞的研究进展[J],北京口腔医学,2006,14(4):289-294
[14]Blau HM,Brazelton TR,Weimann JM.The evolving concept of a stem cell entity of function[J].Cell,2001,105(7):829-841
[15]丁强,贾心善。成体干细胞分化调控研究的新进展[J],日本医学介绍,2004,25(1):39-40
[16]Lakshmipathy U,Verfaillic C.Stem cell plasticity[J].Blood Rev,2005,19(1):29-38
[17]黄海霞,汤学明。成体干细胞多能性研究进展[J],生命科学,2002,14(3):129-135
[18]李建生,刘敬霞。成体干细胞在中枢神经系统疾病修复的效应[J],中国组织工程研究与临床康复2007,Ⅱ(11):2144-2149
[19]Ferrari G,Cusella G,Angelis D,et al.Muscle regeneration by bone marrow derived myogenic progenitors[J].Science,1998,279:1528-30
[20]Theise ND,Nimmakayaly M,Gardner R,et al.Liver from bone marrow stem cells can differentiate into hepatocytes in vivo[J].Nat Med,2000,32(1):11-16
[21]Orlic D,Kajstura J,Chimenti S,et al.Bone marrow cells regenerate infarcted myocarduim[J].Nature,2001,410:701-705
[22]Luo LL.Infertility and sterility[M].Beijing:People Healthy Press.1998:5-6
[23]Smikel CB,Dandekar PV,Schrioch ED,et al.Elevated ovarian follicular fluid stem cell factor concentrations are associated with improved pregnancy rates in vitro fertilization cycles[J].Fertil Steril,1998,69(1):70-72
[24]Smikel CB,Dandekar PV,Schrioch ED,et al.Elevated ovarian follicular fluid stem cell factor concentrations are associated with improved pregnancy rates in vitro fertilization cycles[J].Fertil Steril,1998,69(1):70-72
[25]Tanikawa M,Harada T,Zwabe T,Presence of stem cell factor in follicular fluild and its expression in the human ovary[J].Fertil Steril,2000,73(6):1259-60
[26]Parrott JA,Skinner MK,Kit-ligand and stem cell factor induces primordial follicle development and initiates folliculogenesis[J].Endocrinology 1999,140(9):4262-71
[27]Parrott JA,Skinner MK,kit-ligand action on ovarian stromal cells:effects on the cell recruitment and steroid production[J].Mol Reprod Dev 2000,55(1):55-64
[28]Castrillon DH,Miao L,Kollipara R,et al.Suppression of ovarian follicle activation in mice by the transcription factor FOXO3a[J].Science 2003,301(5630):215-218
[29]Reddy P,Shen L,Ren C,et al.Activation of Akt and suppression of FKHRL1 in mouse and rat oocyte by stem cell factor during follicular activation and development[J].Dev Biol,2005,281(2):160-170
[30]余荣娇,徐斯凡。干细胞因子在精子发生中的作用[J],江西医学院学报2004,44(4):114-116
[31]陈振文,白照岱。胚胎干细胞向生殖细胞的分化研究进展[J],中国男科学杂志2007,21(2):1-5
[32]Young LE.Focus on stem cells in reproduction[J]. Reproduction,2006,132(5):671-672
[33]Zuckerman S,The number of oocytes in the mature ovary[J].Recent Prog.Hom.Res.6:63-108
[34]杨晓菁,姚红军,王玉凤。哺乳动物卵母细胞凋亡的研究进展[J],动物学杂志 2004,39(6):101-106
[35]Hirshfield AN.Development of follicles in the mammalian ovary[J].Int Rev Cytol.1991,124:43-101
[36]罗丽莉,黄菊,傅玉才。内分泌因子,生长因子、细胞因子与卵巢卵泡的发育[J],中国组织工程研究与临床康复 2007,11(19):3821-30。
[37]Joshua Johnson,Jacqueline Canning,Tomoko Kaneko,Germline stem cells and follicular renewal in the postnatal mammalian ovary[J],Nature,2004(428):145-153。
[38]Roger G.Gosden Germline stem cells in the postnatal ovary:is the ovary more like a testis?[J]Human reproduction update,2004(10) 193-195
[39]Bukovsky A,Svetlikova M,Caudle MR.Oogenesis in cultures derived from adult human ovaries[J].Reprod Biol Endocrinol,2005,3(1):17-30
[40]Hubner K,Fumann G,Christenson LK,et al.Derivation of oocytes from mouse embrynic stem cells[J].Science 2003,300(23):1251-56
[41]Jonoathan L,Tilly Commuting the death sentence:how oocytes strive to survive[J],nature,2001(2):838-840
[42]Joshua Johnson,Jessamyn Bagley,Malgorzata Skaznik-Wikiel Oocyte generation in adult mammalian ovaries by putative germ cells in bone marrow and peripheral blood[J],Cell,Vol122:303-315
[43]侯友贤。恶性肿瘤放疗与康复[M],人民军医出版社,2005年06月:20-29
[44]于金明。2005年肿瘤精确放疗进展(修订版)[M],第三届济南国际肿瘤放射治疗新进展暨第二届全国中青年放射肿瘤学术研讨会论文汇编:31-36
[45]王奇璐。肿瘤化疗、放疗201个怎么办(第二版)[M],中国协和医科大学出版社,2004,08:9-11
[46]姚琴,李彦.放疗的副作用及护理[J],国外医学护理学分册,2001,20(3):134-137
[47]孙建衡。妇科恶性肿瘤放疗的几个问题[J],肿瘤学杂志,2006,12(5):357-360
[48]Seymour JF.Ovarian tissue cryoreservation for cancer patients:who is appropreiate?[J].Reprod Fertil,2001(13)81-89
[49]Blumenfeld Z.Preservation of fertily and ovarian fuction and minimalization of chemotherapy associated gonadotoxity and premature ovarian failure:the role of inhibin-A and-B as marks[J].Mol Cell Endorinol,2002,187:93-105
[50]Richardson SJ,Senikas V,Nelson JF.Follicular depletion during the menoarusal transition:evidence for accelerated loss and ultimate exhaustion[J].J.Clin.Endocrinal.Metab.1997(65):1231-37
[51]Feniche IP.Medical preservation of ovarian function before and during radiochemotherapy[J].Gynecol Obstet Fertil,2005,33(9):610-614
[52]Louise D,Bath LE,Anderson RA,et al.Hypothalarmic pituitary ovarian dysfuction after radiotherapic prepubertal ad cranial irradiation for acute leukaemia[J].Hum Reprod,2001,16(9):1838-44
[53]Meirow D,Nugent D.The effects of radiotherapy and chemotherapy on female reproduction[J].Hum Reprod Update,2001,7(6):535-543
[54]Thibaud E,Rodriguez Macias K,Irivin C,et al.Ovarian functino after bone marrow transplantation during childhood[J].Bone Marrow Transplant,1998,21:287-290
[55]Chiarelli AM,Marrett LD,Darlington G,et al.Early menopause and infertility in female after treatment for chindhood cancer diagnosed in 1964-1988 in Ontario,Canada[J].Am JE Epidemiol,1999,150:245-254
[56]Gosden RG,Wade JC,Fraser HM,et al.Impact of congenital hypogonadotropism on the radiation sensitivity of the mouse ovary[J].Hum Reprod,1997,12:2483-2488
[57]Bertram JS.The molecular biology of cancer[J].Mol Aspects Med,2000,21(6):167-233
[58]吴郁,向阳。化疗及放疗对卵巢的影响及预防[J],国外医学妇产科分册,2003,30(3):166-170
[59]Almodin CG,Minguett VC,Meister H,et al.Recovery of natural fertility after grafting of ciyopreserved germinative tissue ewes subjected to radiotherapy[J].Fertil Steril,2004,81(1):160-164
[60]Blumenenfeld Z,Shapiro D,Shteinberg M,et al.Preservation of fertility and ovarian function and minimizing gonado-toxicity in young women with systemic lupus erythematosus treated bu radiotherapy[J].Lupus,2000,9(6):401-405
[61]Pereyra-Pacheco B,Mendez Ribas.Use of GnRH analogs for functional protection of the ovary and preservation of fertility during cancer treatment in adolescent a preliminary report[J],Gynecol Oncol,2001,81(3):391-397
[62]Bath LE,Wallace WH,Shaw MP,et al.Depletion of ovarian reserve in young women after treatment for cancer in childhood:detection by anti-mullertian hormone,inhibin B and ovarian ultrasound[J],Hum Reprod,2003,18(11):2368-74
[63]Domans MM,Demylle D,Martine B,et al.Efficacy of in vitro fertilization after radiotherapy[J],Fertil Steril,2005,83(4):897-901
[64]Berek JS,Hacker NF.Practical Gynecologic oncology[J],Lippincott Willians Wilkins,2004,23(4):119-162
[65]Oktay K,Buyuk E,Veeck L,et al.Embryo development after heterotopic transplantation of cryopreserved ovarian tissue[J],Lancet,2004,363(9412:837-840)
[66]Meirow D.Ovarian injury and mordern options to preserve fertility in female concer patient treated with hign dose radio-chemotherapy for hemato-oncological meoplasias and other cancers[J].Leukema Lymphoma,1999,33(12).65-76
[67]Morice P,Juncker L,Ray A,et al.Ovarian transposition for patients with cervical carcinoma treated by radiosurgical combination[J],Fertil Steril,2000,74:743-748
[68]Blumenfeld Z,Dann E,Avivi I,et al.Fertility after treatment for Hodgkin's disease[J].Ann Oncol,2002,13[suppl 11]:138-147
[69]Morita Y,Perez GI,Paris EOocyte apoptosis is suppressed by disruption of the acid sphingomylinase gene or by sphingosine-1-phosphate therapy[J],Nat Med,2000,6:1109-1114
[70]潘兴华,段连宁,韩毅冰。成体干细胞的可塑性及临床意义[J],Journal of tropical Medicine,2001,1(2):158-163
[71]张静。成体干细胞的应用[J],生物学杂志,2005,22(4):64。
[72]Theise ND,Nimmakayalu M,Gardner T,et al.Liver from bone marrow in human[J].Hepatology,2000,32(11):11-16
[73]刘芊,高敏。骨髓间充质干细胞向肝细胞分化的研究[J].国际消化杂志,2007,27(1):7-10
[74]Clarke DL,Johansson CB,Frisen J,et al.Generalized potential of adult stem cells[J].Science,2002,286:1660-1661
[75]Pittenger MF,Mackay AM,Beck SC,et al.Multilineage potential of adult human mesenchymal stem cells[J],Science,1999,284:143-147
[76]Filip S,Mokr J,Hruska I.Adult stem cells and their importance in cell therapy[J].Folia Biologica,2003,49:9-14
[77]David AP.Current science of regenerative medicine with stem cells[J].Journal of Investive Medicine.2006,54(1):33-35
[78]Vieyra DS,Jackson KA,Goodell MA.Plasticity and tissue regenerative potential of bone marrow-derived cells[J].Stem Cell Rev.2005;1(1):65-69。
[79]潘兴华,张步振。自体骨髓干细胞的临床应用及存在的问题[J],中国医疗前沿,2007,30(15):25-28