Investigation of follicular development and oocyte maturation after cryopreservation and xenograft of newborn mouse ovaries / 生理学报

In order to explore the feasibility of cryopreserving primordial follicles in attaining their developmental competence following freezing and thawing, ovaries from newborn mice were cryopreserved and the thawed ovaries were xenografted into kidney capsules of adult female mice. Ovaries were isolated from newborn B6C2F(1) female mice, infiltrated by Leibovitz 15 (L-15) medium containing 10% (V/V) fetal bovine serum (FBS) and 1.5 mol/L dimethylsulfoxide (DMSO), and then packed into 0.25 ml plastic straws. The ovaries contained in straws were frozen under nitrogen vapour at -40 degrees C in Cryocell 1200 programmable freezer, and stored in liquid nitrogen for periods ranging from 1 week to 6 months. Upon thawing, the straws were dipped into room temperature water for 10~20 s, after which the ovaries were collected and washed in L-15 buffer containing 10% (V/V) FBS without DMSO to remove cryoprotectant. The thawed ovaries were transplanted into kidney capsules of 8~12-week old adult B6C2F(1) female recipient mice by two protocols, with either 1 or 2 ovaries in each capsule. Upon withdrawal after at least 14 d of transplantation, only 45.00% (72/160) of the ovaries were recovered from 40 recipients transplanted with 2 ovaries in each capsule, compared to 82.50% (33/40) in 20 recipients with only 1 ovary in each capsule. The grafted ovaries exhibited similar follicular developmental progression to that of natural ovaries. There were antral follicles present in the transplanted ovaries on day 14, whose number increased more substantially on day 19 after transplantation. Following stimulation of the recipient mice with 10 IU PMSG on day 19 after xenografting, follicles further developed to preovulatory stage with appearance of cumulus oocytes and enlarged antrum. Oocytes from these fully grown antral follicles were collected and matured in vitro in modified essential medium-alpha (MEMalpha). After 16~17 h of culture, 40.90% of the oocytes exhibited germinal vesicle breakdown (GVBD) and among which 89.02% proceeded to the metaphase II (MII) stage as indicated by exclusion of the first polar body. The remaining oocytes were further cultured and 50.83% of which initiated GVBD by 20~21 h of culture, but only 21.40% of which proceeded to MII. The above results demonstrated that the primordial follicles in newborn mouse ovaries were capable of sustaining freezing and thawing, and reinitiating development following xenograft into kidney capsule in adult recipient female mice. Production of mature oocytes from such re-developed follicles following gonadotrophin priming and the subsequent oocyte in vitro maturation implied immense prospect of application of this method to preserve female germ cells, conserve endangered species, establish animal gene stock, and utilize oocytes in assisted reproductive techniques.

Treatment of nonneoplastic epithelial disorders of skin and mucosa of vulva with focused ultrasound / 中华妇产科杂志

Objective To explore the clinical efficacy of focused ultrasound for patients with white lesions of the vulva,as well as its safety and feasibility.Methods Clinical data of 941 patients with white lesions of the vulva treated with focused ultrasound from June 2003 to December 2005 were retrospectively reviewed.The mean age of the patients was 40.8 years(18-70 years)and the median course of the disease was 6.2 years(3 months-45 years).Meanwhile,pathological diagnosis was performed in all the patients before treatment,in which 498 cases were squamous hyperplasia,342 cases were lichen sclerosus and 101 cases were lichen sclerosus with squamous hyperplasia.Patients were followed up and therapeutic effects of focused ultrasound was evaluated at 6 and 12 months after the treatment,respectively.The symptoms of pruritus in the vulva and the changes in the color and elasticity of the vulvar lesions were observed.Results Of all the patients,900 were followed up after the treatment,and the ratio of effectiveness was 94.9%.Only 46 patients(5.1%)had no response to the therapy.Of the effective patients,434 cases were completely cured(48.2%),and 420 cases were improved(46.7%).Pruritus of vulva recurred in 101 patients (11.2%)one year after treatment;however,these patients still had a response to the second or third treatment.Conclusions Focused ultrasound therapy is a highly effective instrument in treatment of white lesions of the vulva.It can not only relieve the symptoms of itching,but is also helpful in recovering the color and elasticity of the vulva.