Objective To evaluate the impact of slow-freezing process on human ovarian tissue with the standard cryopreservation-thawing protocol of Fertility Protection Center of Beijing Obstetrics and Gynecology Hospital,Capital Medical University.Methods Ovarian tissues of 12 patients were divided into fresh ovarian tissue group(fresh group)and freezing-thawing ovarian tissue group(F-T group).The freezing-thawing protocol was the standard protocol in our center.The number and activity of follicle were examined with Hematoxylin-eosin(HE)staining and calcein-AM(calcein acetoxymethylester)staining,and the proliferation and apoptosis was evaluated with the immunohistochemical staining of Ki-67 and caspase-3.The expressions of apoptosis-related proteins such as caspase-3,bax and FasL between the two groups were compared with Western blotting.Results There were no statistically significant differences in follicle counting and follicle activity in ovarian tissues pre-and post-freezing-thawing(P>0.05),and the positive rate of Ki-67 in ovarian tissues after freezing-thawing was significantly lower than that in fresh ovarian tissues(P<0.05),and there was no statistically significant difference in the positive rate of caspase-3 between the two groups(P>0.05).The expression of caspase-3 protein in ovarian tissues after freezing-thawing was significantly higher than that in fresh ovarian tissues(P<0.05),while the expressions of other apoptosis-related proteins such as bax and FasL were not significantly different(P>0.05).Conclusion The standard cryopreservation-thawing regimen in our center can effectively maintain the follicle number,morphology,and activity in ovarian tissues.After freezing and thawing,the cell proliferation level is decreased.The expression of apoptosis-related proteins such as bax and FasL are not increased,and the expression of caspase-3 is relatively increased.These results suggest our freezing-thawing regimen is good for human ovarian tissue.

Female infertility is recognized as a global public health issue by the World Health Organization.Female fertility remodeling includes ovarian function reconstruction and uterus/endometrium reconstruction,etc.It is emerging as a hot technology since it is ready to prepare autologous platelet-rich concentrate and it is safer and more acceptable in autologous application.It plays an important role in regenerative medicine,and it is currently widely applied in maxillofacial and plastic surgery,dermatology and other clinical practices.This article mainly reviews the progresses of the application of autologous platelet-rich concentrate in female fertility remodeling.

Objective To evaluate the impact of slow-freezing process on human ovarian tissue with the standard cryopreservation-thawing protocol of Fertility Protection Center of Beijing Obstetrics and Gynecology Hospital,Capital Medical University.Methods Ovarian tissues of 12 patients were divided into fresh ovarian tissue group(fresh group)and freezing-thawing ovarian tissue group(F-T group).The freezing-thawing protocol was the standard protocol in our center.The number and activity of follicle were examined with Hematoxylin-eosin(HE)staining and calcein-AM(calcein acetoxymethylester)staining,and the proliferation and apoptosis was evaluated with the immunohistochemical staining of Ki-67 and caspase-3.The expressions of apoptosis-related proteins such as caspase-3,bax and FasL between the two groups were compared with Western blotting.Results There were no statistically significant differences in follicle counting and follicle activity in ovarian tissues pre-and post-freezing-thawing(P>0.05),and the positive rate of Ki-67 in ovarian tissues after freezing-thawing was significantly lower than that in fresh ovarian tissues(P<0.05),and there was no statistically significant difference in the positive rate of caspase-3 between the two groups(P>0.05).The expression of caspase-3 protein in ovarian tissues after freezing-thawing was significantly higher than that in fresh ovarian tissues(P<0.05),while the expressions of other apoptosis-related proteins such as bax and FasL were not significantly different(P>0.05).Conclusion The standard cryopreservation-thawing regimen in our center can effectively maintain the follicle number,morphology,and activity in ovarian tissues.After freezing and thawing,the cell proliferation level is decreased.The expression of apoptosis-related proteins such as bax and FasL are not increased,and the expression of caspase-3 is relatively increased.These results suggest our freezing-thawing regimen is good for human ovarian tissue.

Female infertility is recognized as a global public health issue by the World Health Organization.Female fertility remodeling includes ovarian function reconstruction and uterus/endometrium reconstruction,etc.It is emerging as a hot technology since it is ready to prepare autologous platelet-rich concentrate and it is safer and more acceptable in autologous application.It plays an important role in regenerative medicine,and it is currently widely applied in maxillofacial and plastic surgery,dermatology and other clinical practices.This article mainly reviews the progresses of the application of autologous platelet-rich concentrate in female fertility remodeling.

Objective To investigate the effect of Sclareol(SCL)on LX-2 hepatic stellate cell activation and CCl4-induced liver fibrosis in mice,and to further explore its mechanism.Methods A total of 40 Kunming mice were randomly divided into healthy group,model group(10%CCl4)and SCL administration group,and silybin positive control group(10%CCl4+100 mg·kg-1 Silybin),and SCL administration group was divided into low SCL(10%CCl4+20 mg·kg-1 SCL)and high dose group(10%CCl4+40 mg·kg-1 SCL).Mice in all groups were intraperitoneally injected with 10%olive oil-diluted CCl4 three times a week for four weeks,except for the healthy group.Starting from the third week,the dosing group was given different doses of SCL by gavage daily,and the positive control group was given silybin daily,and the mice were sacrificed and serum and liver tissue were collected after four weeks.In whole animal experiments,biochemical kits were used to detect the changes in the serum levels of glutamate aminotransferase(ALT)and aspartate aminotransferase(AST)in mice with liver fibrosis.Hematoxylin-eosin(HE),Sirius Red and Masson staining were used to detect microstructural changes and collagen deposition in liver tissues.Immunohistochemistry was used to detect the expression of fibrosis marker proteins α-smooth muscle actin(α-SMA)and fibrous collagen I.in liver tissues.In vitro,LX-2 human hepatic stellate cells were used for normal culture in the blank group,and the activation of LX-2 hepatic stellate cells was induced by transforming growth factor-β1(TGF-β1)in the model group,and the SCL administration group was divided into SCL low-dose group(5 ng·mL-1 TGF-β1+10 μmol·L-1 SCL)and high-dose group(5 ng·mL-1 TGF-β1+20 μmol·L-1 SCL).Subsequently,Transwell and EdU assays were used to detect the effects of SCL on the migration and proliferation of LX-2 cells.The expression of fibrosis marker proteins α-SMA and Collagen I.affected by SCL was detected by immunofluorescence.Western blot was used to detect the expression of related proteins in TGF-β/Smad pathway.Results In animal experiments,compared with the model group,SCL could significantly improve the liver function indexes and liver histopathological changes in liver fibrosis model mice.In addition,in vitro cell experiments,compared with the model group,SCL can effectively inhibit the migration and proliferation of hepatic stellate cells and inhibit their activation.Further studies showed that compared with the model group,SCL significantly up-regulated the expression of Smad7 protein and significantly down-regulated the phosphorylation levels of Smad2 and Smad3 proteins.Conclusion SCL has a significant alleviating effect on CCl4-induced liver fibrosis and TGF-β1-induced LX-2 activation in mice,and the mechanism may be related to the regulation of TGF-β/Smad pathway.

Objective To investigate the effect of Sclareol(SCL)on LX-2 hepatic stellate cell activation and CCl4-induced liver fibrosis in mice,and to further explore its mechanism.Methods A total of 40 Kunming mice were randomly divided into healthy group,model group(10%CCl4)and SCL administration group,and silybin positive control group(10%CCl4+100 mg·kg-1 Silybin),and SCL administration group was divided into low SCL(10%CCl4+20 mg·kg-1 SCL)and high dose group(10%CCl4+40 mg·kg-1 SCL).Mice in all groups were intraperitoneally injected with 10%olive oil-diluted CCl4 three times a week for four weeks,except for the healthy group.Starting from the third week,the dosing group was given different doses of SCL by gavage daily,and the positive control group was given silybin daily,and the mice were sacrificed and serum and liver tissue were collected after four weeks.In whole animal experiments,biochemical kits were used to detect the changes in the serum levels of glutamate aminotransferase(ALT)and aspartate aminotransferase(AST)in mice with liver fibrosis.Hematoxylin-eosin(HE),Sirius Red and Masson staining were used to detect microstructural changes and collagen deposition in liver tissues.Immunohistochemistry was used to detect the expression of fibrosis marker proteins α-smooth muscle actin(α-SMA)and fibrous collagen I.in liver tissues.In vitro,LX-2 human hepatic stellate cells were used for normal culture in the blank group,and the activation of LX-2 hepatic stellate cells was induced by transforming growth factor-β1(TGF-β1)in the model group,and the SCL administration group was divided into SCL low-dose group(5 ng·mL-1 TGF-β1+10 μmol·L-1 SCL)and high-dose group(5 ng·mL-1 TGF-β1+20 μmol·L-1 SCL).Subsequently,Transwell and EdU assays were used to detect the effects of SCL on the migration and proliferation of LX-2 cells.The expression of fibrosis marker proteins α-SMA and Collagen I.affected by SCL was detected by immunofluorescence.Western blot was used to detect the expression of related proteins in TGF-β/Smad pathway.Results In animal experiments,compared with the model group,SCL could significantly improve the liver function indexes and liver histopathological changes in liver fibrosis model mice.In addition,in vitro cell experiments,compared with the model group,SCL can effectively inhibit the migration and proliferation of hepatic stellate cells and inhibit their activation.Further studies showed that compared with the model group,SCL significantly up-regulated the expression of Smad7 protein and significantly down-regulated the phosphorylation levels of Smad2 and Smad3 proteins.Conclusion SCL has a significant alleviating effect on CCl4-induced liver fibrosis and TGF-β1-induced LX-2 activation in mice,and the mechanism may be related to the regulation of TGF-β/Smad pathway.

In this paper,we explored to develop a new concept of computer-and internet-based learning and training method in medicine,especially in obstetrics and gynecology,which is named as Virtual Academy of Women'Health(VA).Especially in the times of infectious disease pandemics worldwide,learning at home rather than in big lectures hall,might be necessary and practical as never before.The VA is based on worldwide knowledge in medicine—free accessible on the internet—in terms of homepages,video and audio platforms,scientific papers,medical books,and different guidelines.A collection of different video-clips in various fields of women's health can assist the student or doctor in understanding the symptoms,diagnostics,and treatment of various diseases.There are two major targets of it—one is online education,and one is testing the knowledge by simulation of clinical cases.

In recent years,with the rapid development of medical research,cancer diagnosis and treatment technology have significantly improved young cancer patient's survival rate.Anticancer therapy such as chemotherapy,radiother-apy,or hematopoietic stem cell transplantation can lead to premature ovarian insufficiency.The endocrine and reproductive function of the ovary is critical to women's physical and mental health.Ovarian tissue cryopreser-vation and transplantation can protect not only female fertility but also preserve ovarian endocrine function.This paper interprets the guidelines for ovarian tissue cryopreservation and transplantation issued by the Chinese Society of Gynecological Endocrinology affiliated to the International Society of Gynecological Endocrinology.The purpose of this guideline's interpretation is to promote more medical workers to understand the technology of ovarian tissue cryopreservation and transplantation,which can provide patients with more choices of fertility protection methods and improve their quality of life.

Objective:To preserve fertility and endocrine function of an endometrial carcinoma patient by ovarian tissue cryopreservation and transplantation.Methods:Part of the ovary was harvested and cryopreserved when surgery for a 33-year-old woman who was diagnosed with endometrial carcinoma. After the cancer reached remission, the patient underwent cryopreserved ovarian tissue transplantation. The endocrine hormone, follicle growth, and menopausal syndromes were detected.Results:One month after the surgery of transplantation, the modified Kupperman scores decreased from 12 to 2, and the menopausal syndrome disappeared. Two months after the operation of transplantation, the concentration of follicle-stimulating hormone (FSH) decreased to 21.78 IU/L, the level of estradiol increased from 13.76 ng/L to 65.55 ng/L, and follicle growth was observed by ultrasound. Four months after the surgery of transplantation, the level of FSH decreased to 6.26 IU/L, the level of estradiol increased to 112.22 ng/L. Nine months after the operation of transplantation, the level of estradiol was 118.14 ng/L, the level of progesterone increased to 8.96 μg/L, and two corpus luteum after ovulation were detected by ultrasound.Conclusion:Two months after the surgery of transplantation, the ovarian function has returned to normal, which demonstrated the success of transplantation of frozen-thawed ovarian tissue.

Objective:To preserve fertility and endocrine function of an endometrial carcinoma patient by ovarian tissue cryopreservation and transplantation.Methods:Part of the ovary was harvested and cryopreserved when surgery for a 33-year-old woman who was diagnosed with endometrial carcinoma. After the cancer reached remission, the patient underwent cryopreserved ovarian tissue transplantation. The endocrine hormone, follicle growth, and menopausal syndromes were detected.Results:One month after the surgery of transplantation, the modified Kupperman scores decreased from 12 to 2, and the menopausal syndrome disappeared. Two months after the operation of transplantation, the concentration of follicle-stimulating hormone (FSH) decreased to 21.78 IU/L, the level of estradiol increased from 13.76 ng/L to 65.55 ng/L, and follicle growth was observed by ultrasound. Four months after the surgery of transplantation, the level of FSH decreased to 6.26 IU/L, the level of estradiol increased to 112.22 ng/L. Nine months after the operation of transplantation, the level of estradiol was 118.14 ng/L, the level of progesterone increased to 8.96 μg/L, and two corpus luteum after ovulation were detected by ultrasound.Conclusion:Two months after the surgery of transplantation, the ovarian function has returned to normal, which demonstrated the success of transplantation of frozen-thawed ovarian tissue.