| BackgroundIntrauterine adhesions (IUA) also named Asherman snydrome, and first described by Fritsch. In1948, Ashenman originally defined IUA as a consequence of trauma to the endometrium,producing partail or complete obliteration in the uterine cavity and/or the cervical canal, resulting in conditions such as menstrual abnormalities, infertility, and recurrent pregnancy loss. The morbility of IUA is accounted for5.1%of secondary amenorrhea, and4.8%of female infertility, and37.6%of post-abortion curettage. With the frequency of intrauterine manipulation and the popularity of hysteroscopic surgery, the incidence of IUA is increasing gradually during the recent years. Trend of incidence age of patients with IUA gradually gets younger and IUA has become the second cause of fenmale secondary infertility. The cure rate and pregnancy rate of IUA were not obviously increased from lack of effective therapy. The recurrence rate of patients with mild and severe IUA are33.3%and66.7%respectively. Obstetric complications resulting from IUA are the threatens to female’s reproductive health, such as infertility, repeated miscarriage, premature delivery, placenta previa, and placenta accreta. IUA treatment is focused by researchers, and how to solve female fertility dysfuntion is hot spot in clinic. The aim of treatment for IUA is to restore the size and shape of the uterine cavity, prevent recurrence of the adhesion, promote tne repair and regeneration of the destroyed andometrium, and restore normal reproductive functions. The three steps of IUA treatment are hysteroscopic adhesiolysis, insertion of intrauterine device and hormonal treatments. The therapeutic schedule has long treatment cycle, low cure rate and pregnancy rate, and easy to relapse. The risk of breast and endometrial cancer is increased for taking orally large dose estrogen, and estrin treatment is ineffective to severe IUA.To date, researchers agreed that the disturbance of endometrial repair might be an important mechanism for IUA. The disturbance of endometrial repairment followed by post-abortion curettage or other intrauterine operation, formed scar and adhesions under the pathological circumstances. In recent years, endometrial stem cells to be confirmed provided new theoretical basis for various gynecological disease involved with endometrium abnormal growth or repairment, such as endometrial cancer and endometriosis. Thin endometrium and IUA might be associated with the reduction and absence of endometrial stem cells.Bone marrow mesenchymal stem cells are a population of multipotent cells and ideal source of stem cells for tissue engineering. BMSCs were attracted much attention with advantages of self-renewing, pluripotent differentiation, and avoiding the immune rejection and ethics, moral problems, especially to repair damaged tissue. Transplantation of BMSCs can be used for treating various traumatic diseases, such as myocardial injury caused by myicardial infarction and dilated cardiomyopathy, acute kidney injury, and bone, cartilage and tendons defects. However, studies on BMSCs repairing endometrial injury involved with IUA are not available.In the present study, we separated, cultured the BMSCs in vitro, and labeled with PKH26. BMSCs are induced into endometrial epithelial cells or stromal cells in vitro. After entablishment of New Zealand white rabbit model for IUA using mechanical and infectious injury, the labeled BMSCs were transplanted directly into the uterus. The survival of the cells, the changes of endometrium, and expression of gene and protein of endometrium were detected.Chapter1Separation, culture, identification and label of New Zealand white rabbit bone mesenchymal stem cellsObjective:To eatsblish the method of separation, culture of the New Zealand white rabbit bone marrow mesenchymal stem cells in vitro, and study the biological characteristica and phenotype of the cultured cells. PKH26was compared cellular proliferation and phenotype with unlabeled cells.Methods:1. The femus and tibia were removed from female New Zealand white rabbits after euthanasia with overdose of anethesia. The bone mesenchymal stem cells were separated and purified whole marrow adherent method,and then cultured with DMEM/F12media contained with10%fetal bovine serum (FBS). At confluence, the cells were harvested for passage with0.25%trypsin containing0.02%EDTA.2. The expressions of CD44, CD45and CD90in MSCs were analyzed by flow cytometer (FCM).3. PKH26was used for the label of BMSCs and the immediate efficiency of PKH26was quantified. The growth curves were delineated to identify whether the PKH26influenced on the cell growth characteristics in vitro. Furthermore, the surface markers (CD44, CD45, and CD90) of labeled BMSCs were tested through FCM and compared with unlabeled BMSCs.Results: 1. The primary cultured BMSCs were spindle-shaped and adherent and round24hours after innoculation. Most of the cells were adherent and fusiform-shaped72hours after innoculation. The cells were passaged after9-10days. The third passage of BMSCs were morphologically resembled fibroblast.2. Ninety-five percent of the cultured cells expressed CD44and CD90but CD45, as were identified BMSCs.3. The immediate efficiency of PKH26labeling was100%. There was no statistical difference in cell apoptotic tatio between labeled BMSCs and unlabeled BMSCs. The growth curves of labeled BMSCs were similar with that of unlabeled BMSCs. The PKH26didn’t influence the expressions of surface markers of rabbit BMSCs.Conclusions:Purified BMSCs can be obtained by whole marrow adherent method, and were positive for CD44and CD90, negative for CD45. The cultured BMSCs were uniform and stable, and could be used for tissue engineering. PKH26can be serve as a label of rabbit BMSCs for tracing due to its high and stable label efficiency.Chapter2The in vitro study of differentiation of bone marrow mesenchymal stem cells into endometrial cells in rabbitsObjective:To investigate the differentiation of BMSCs into endometrial cells under specified conditions and its regulatory mechanism.Methods:1. The endometrial conditioned medium was prepared.2. Experiment grouping design:(1) control group:complete medium;(2) ECM group:complete medium containing20%ECM;(3) ECM+E-8group:complete medium containing20%ECM and10-8mol/L17β-E2;(4) ECM+E-7group: complete medium containing20%ECM and10-7mol/L17β-E2;(5) ECM+E-6group:complete medium containing20%ECM and10-6mol/L17β-E2;(6) ECM+E-5group:complete medium containing20%ECM and10-5mol/L17β-E2.3. The RNA of BMSCs was extracted after5days, and the expressions of CK18, ESR1and VIM were tested by FQ-PCR.4. The protein of BMSCs was extracted after5days, and the expressions of CK, VIM and ESR1proteins were tested by Western blot.5. The expression of the epithelial marker CK protein was tested by immunofluorescence.Results:1. The results of FQ-PCR were as follows:(1) The genetic expression of CK18had significant difference among groups (F=125.558, P=0.000), and the descending order of its expression was ECM(7.64), ECM+E-6(6.15), ECM+E-7(5.62), ECM+E-8(4.70), ECM+E-5(1.49) and control group(1.00). There was no significant difference of genetic expression of CK18between ECM+E-5and control group, and was significant difference between the rest four groups and control group (PECM=0.000, PE-8=0.000, PE-7=0.000, PE-6=0-000, PE-5=0.168). A strong negative correlation between the genetic expression of CK18and the concertation of17β-estradiol was observed (rs=-0.615, P=0.000).(2) There was significant difference of genetic expression of ESR1among groups (F=191.470, P=0.000). The descending order of ESR1expression was ECM(1.45), ECM+E-6(1.38), ECM+E-8(1.37), ECM+E-7(1.36), control(1.00) and ECM+E-5group(0.73). The ESR1expression in all experimental groups were significant difference compared with control group(P=0.000). A strong negative correlation between the genetic expression of ESR1and the concertation of 17β-estradiol was observed (rs=-0.729, P=0.000).(3) It was significant difference that the level of VIM genetic expressed among groups (F=911.786, P=0.000). The descending order of VIM expression was ECM(8.22), ECM+E-5(7.29), ECM+E-7(6.15), ECM+E-8(4.67), ECM+E-6(2.33) and control group(1.00). The VIM expression in all experimental groups were significant difference compared with control group(P=0.000). A strong positive correlation between the genetic expression of VIM and the concertation of17β-estradiol was observed (rs=0.611, P=0.000).2. Quantitiative analysis of the results of Western blot were as follows:(1) The protein expression of CK in experimental group was significantly higher than that of control group (F=478.067, P=0.000). The descending order of CK protein expression was ECM+E-5(0.92), ECM+E-6(0.90), ECM+E-7(0.88), ECM(0.84), ECM+E-8(0.58) and control group(0.27). The CK expression in all experimental groups were significant difference compared with control group(P=0.000). A strong positive correlation between the protein expression of CK and the concertation of17β-estradiol was observed (rs=0.725, P=0.000).(2) The protein expression of ESR1in experimental group was significantly higher than that of control group (F=659.714, P=0.000). The descending order of ESR1protein expression was ECM(0.75), ECM+E-7(0.73), ECM+E-5(0.35), control group(0.17), ECM+E-8(0.14) and ECM+E-6group(0.11). There was no significant difference of genetic expression of ESR1between ECM+E-8and control group, and was significant difference between the rest four groups and control group (PECM=0.000, PE-8=0.101,-PE-7=0.000, PE-6=0.002, PE-5=0.000). No correlation between the protein expression of ESR1and the concertation of17β-estradiol was observed(rs=0.017, P=0.929).(3) The protein expression of VIM in experimental group was significantly higher than that of control group (F=909.869, P=0.000). The descending order of ESR1protein expression was ECM+E-6(0.96), ECM(0.92), ECM+E-5(0.91), ECM+E-8(0.85), ECM+E-7(0.77) and control group(0.14). The VIM expression in all experimental groups were significant difference compared with control group(P=0.000). A positive correlation between the protein expression of VIM and the concertation of17β-estradiol was observed (rs=0.382, P=0.037).3. IF studies showed positive CK staining observed in experimental group.Conclusions:1. Rabbit BMSCs can differentiate into endometrial cells induced by endometrial conditioned medium and estradiol. The levels of CK18, VIM and ESR1mRNA, and the protein expression of CK, VIM and ESR1of differentiated BMSCs were significantly increased than those of undifferentiated cells. It was suggested that BMSCs had the potential to differentiate into endometrial glandular epithelial cells and stromal cells.2. The endometrial microenvironment simulated by endometrial conditioned medium plays a key role in the process of differentiation, and estradiol plays a regulatory role in the process.Chapter3The study of creation of a female New Zealand white rabbit model for intrauterine adhesionsObjective:To explore the creation of a New Zealand white rabbit model for IUA through mechanical, infectious and dual injury respectively according to two major causes of IUA:trauma and infection. Through this study, we hope to provide an ideal animal model for the study of the exact pathogenesis of and therapy for this difficult clinical condition.Methods: 1. The adult female New Zealand white rabbits were used to establish the model. Preparation of lipopolysaccharide surgical suture was used the10-0medical sterile surgical suture the day before use in the model.2. Sixty-four female rabbits were randomly divided into four groups:control (16), mechanical injury (16), infectious injury (16), and dual injury (16). Three methods were applied to esta blish the model:uterine curettage, uterine cavity left alone, lipopolysaccharide surgical suture in place for48h, and suture retention for48h after curettage.3. Histologic changes were monitored at0,24,48, and72h and7,14, and28d after operation, and the number of glands were counted by HE stain.4. Endometrial fibrosis was observed by masson stain, and the area tatio of endometrial fibrosis was analysised by IPP software.Results:1. After injury caused by three methods, the surface epithelium was denuded; scattered basal glands remained only in some deeper stromal fold regions. The stroma was bare and thinned; even the full thickness of the endometrium disappeared in the severely injured areas. Extensive recent hemorrhage and neutrophil infiltration were evident in the stroma. The capillaries were ruptured and had bled. Epithelial debris and red blood cells were visible in the uterine cavity. One week after the mechanical and infectious injury, the surface was completely lined by epithelial cells, and no areas of denudation persisted. Beneath the regenerating epithelium, only a few glands could be distinguished. The stroma prominently showed focal congestion and leukocytic infiltration. One week after the dual injury, the uterus was vertically cut, and on the axial view, some highly fibrous adhesions occurred in the uterine cavity. Microscopically, the fibrous adhesions thickened, and scarce rounded glands were present beneath the epithelial layer. 2. The ratios of the area with endometrial fibrosis to total endometrial area were calculated before injury and7d after injury, and the dual injury group had the highest ratio (P=0.000).3. The number of endometrial glands per high-power field was also counted, and the dual injury group had the fewest of the four groups (P=0.000), whereas the mechanical injury group had fewer than the normal group (P=0.030).4. The success rate of establish rabbit model dual injury was the highest among three groups (χ2=11.000, P=0.012).Conclusions:The method of dual injury can cause the irreversible injury of endometrium, scarring repair and fibrosis adhesions, so as to establish a stable rabbit IUA model.Chapter4The in vivo study of repair effect of bone marrow mesenchymal stem celts transplantation combined with estrin on endometrial injuryObjective:To establish a rabbit model of intrauterine adhesions, and to investigate the influence of rabbit bone marrow mesenchymal stem cells transplantation combined with estradiol on injury endometrium of intrauterine adhesions in vivo and its possible mechanisms.Methods:1. Twelve-week old female New Zealand white rabbits were divided into five groups, namely the control group, model group, transplantation group, estrin group and combined treatment group.2. The rabbits model were established by dual injury. The second passage BMSCs marked with PKH26were served as transplanted cells. At pre-treatment,1, 2. and4weeks after treatment,3rabbits from each group were sacrificed to remove the bilateral uteri.3. The sections of the uteri were examined by HE and masson staining to observe the morphology of endometrium.4. The frozen-section of the uteri were examined using fluorescent microscope to track the injected PKH26-labeled BMSCs.5. Serum estrogen was measured by ELISA.6. The genetic level of CK18, VIM, ESR1and TFG-B1of uteri tissues were tested by FQ-PCR.7. The protein quantification of CK and ESR1of uteri tissues were analysised by Western blot.8. The protein expression of CK, VIM, ESR1and TFG-β1of uteri endometrium were examined by IHC.Results:1. The results of HE staining were showed as follow:since1week after treatment, the number of endometrial glands was increased in the three treatment groups, and the new glands were round or oval.(1) Before treatment, there was significant difference among all groups (F=52.074,P=0.000), the number of glands in the latter4groups were all significantly lower than that of control group (P=0.000). There were no significant difference in transplantation group (P=0.482) and estrin group (P=0.723) compared with combined group.(2) One week after treatment, there was significant difference among all groups (F=10.287, P=0.001), the number of glands in the latter4groups were all significantly lower than that of control group (Pmodel=0.000,Ptransplantation=0.001, Pestrin=0.001,Pcombined=0.0002). There were no significant difference in transplantation group (P=0.482) and estrin group (.P=0.723) compared with combined group.(3) Two weeks after treatment, there was significant difference among all groups (F=16.016, P=0.000). there was no significant difference of the number of glands between combined and control group, and the rest three experimental groups were still significantly lower than control group (Pmodel=0.000, Ptransplantation=0.005, Pestrin=0.003, Pcombined=0.124). Among three treatmental groups, there was no significant difference in transplantation group(P=0.079), and was significant difference in estrin group(P=0.049) compared with combined group respectively.(4) Four weeks after treatment, there was significant difference among all groups (F=12.426, P=0.001). there were no significant difference of the number of glands in transplantation and estrin groups compared with control group, and the rest two experimental groups had significant difference (Pmodel=0.002, Ptransplantation=0.411, Pestrin=0-146, Pcombined=0.028). Among three treatmental groups, there was no significant difference in transplantation group(P=0.006) and estrin group(P=0.002) compared with combined group respectively.2. The results of masson staining were showed as follow:the collagen fibers in the endometrial stroma were decreased.(1) Before treatment, there was significant difference among all groups (F=6.746, P=0.007), the number of glands in the latter4groups were all significantly lower than that of control group (Pmodel=0.002, Ptransplantation=0.002, Pestrin=0.002, Pcombined=0.004). There were no significant difference in transplantation group (P=0.629) and estrin group (P=0.688) compared with combined group. (2) One week after treatment, there was significant difference among all groups (F=7.760, P=0.004). The combined group was no significant difference, and the rest three groups had significant difference compared with control group (Pmodel=0.000, Ptransplantation=0.014, Pesrin=0.013,Pcombined=0.138). There were no significant difference in transplantation group (P=0.205) and estrin group (P=0.191) compared with combined group.(3) Two weeks after treatment, there was significant difference among all groups (F=14.045, P=0.000). The transplantation and combined group were no significant difference, and the rest three groups had significant difference compared With Control group (Pmodel=0.000,Ptransplantation=0.918, Pestrin=0.002, Pcombined=0.792). Among three treatmental groups, the transplantation group(P=0.714) was no significant difference and estrin group (P=0.001) had significant difference compared with combined group respectively.(4) Four weeks after treatment, there was significant difference among all groups (F=31.728, P=0.000). The model group had significant difference, and the rest three groups were no significant difference compared with control group (Pmodel=0.000, Ptransplantation=0.342,Pestrin=0.980, Pcombined=0.670). Among three treatmental groups, the transplantation group(P=0.589) and estrin group (P=0.688) were no significant difference compared with combined group respectively.3. Seven days after transplanting, red fluorescence was observed in BMSCs transplantation and combined treatment group under the fluorescence microscope, until twenty-one days after transplantation. There was no red fluorescence observed in the other groups, cervix, vagina and ovary. The red fluorescence was observed mainly near the endometrial epithelium compared with the blue fluorescence staining the nucleus by DAPI. 4. The analysis of serum E2showed as follows:(1) The concentration of serum E2was almost alike between groups before treatment(F=0.023, P=0.999). It explained that the ovarian function was not damaged at the process of establishing animal model.(2) One week after treatment, there was significant difference among all groups (F=67.495, P=0.000). The estrin and combined group had significant difference, and the rest two groups were no significant difference compared with control group (Pmoder=0.943,Ptransplantation0.960, Pestrin=0.000, Pcombined=0.000). Among three treatmental groups, the transplantation group(P=0.000) had significant difference and estrin group (P=0.224) was no significant difference compared with combined group respectively.(3) Two weeks after treatment, there was significant difference among all groups (F=44.854, P=0.000). The estrin and combined group had significant difference, and the rest two groups were no significant difference compared with control group (Pmodel=0.599, Ptransplantat=0.932, Pestrin=0.000, Pcombined=0.000). Among three treatmental groups, the transplantation group(P=0.000) had significant difference and estrin group (P=0.939) was no significant difference compared with combined group respectively.(4) Four weeks after treatment, there was significant difference among all groups (F=10.093, P=0.002). The combined group had significant difference, and the rest three groups were no significant difference compared with control group (Pmodel=0.802, Ptransplantation=0.635, Pestrin=0.094, Pcombined=0.001). Among three treatmental groups, the transplantation group(P=0.041) and estrin group (P=0.010) had significant difference compared with combined group respectively.5. The results of the level of mRNA were showed as follows: (1) The analysis of CK18gentic level①Before treatment, the CK18genetic level of uterine tissue in experimental group had significant difference compared with control group (Pmode;=0.000, Pransplantation=0.000, Pestrin=0.000, Pcombined=0.000). Among three treatmental groups, the transplantation group(P=0.835) and estrin group (P=0.776) were no significant difference compared with combined group respectively.②One week after treatment, the CK18genetic level of uterine tissue in model and transplantation group had significant difference, and the rest two groups were no significant difference compared with control group (Pmodel=0.002, Pransplantation=0-000, Pestrin=0.377, Pcombined=0-434). Among three treatmental groups, the transplantation group(P=0.000) had significant difference and estrin group (P=0.915) was no significant difference compared with combined group respectively.③Two weeks after treatment, the CK18genetic level of uterine tissue in transplantation group had significant difference, and the rest three groups were no significant difference compared with control group (Pmodel=0.197, Pransplantation=0.008, Pestrin=0.062, Pcombined=0.119). Among three treatmental groups, the transplantation group(P=0.180) and estrin group (P=0.101) were no significant difference compared with combined group respectively.④Four weeks after treatment, the CK18genetic level of uterine tissue in model and combined group had significant difference, and the rest two groups were no significant difference compared with control group (Pmodel=0.027, Ptransplantation=0.110, Pestrin=0.110,Pcombined=0-007). Among three treatmental groups, the transplantation group(P=0.252) and estrin group (P=0.351) were no significant difference compared with combined group respectively.(2) The analysis of VIM gentic level①Before treatment, the VIM genetic level of uterine tissue in all experimental groups were significant higher than that in control group (Pmodel=0.026, Ptransplantation=0.008, Pestrin=0.024,Pcombined=0.010). Among three treatmental groups, the transplantation group(P=0.868) and estrin group (P=0.630) were no significant difference compared with combined group respectively.②One week after treatment, the VIM genetic level of uterine tissue in model and combined group had significant difference, and the rest two groups were no significant difference compared with control group (Pmodel=0.027, Ptransplantation=0.110,Pestrin=0.110, Pcombined=0.007). Among three treatmental groups, the transplantation group(P=0.764) was no significant difference and estrin group (P=0.038) had significant difference compared with combined group respectively.③Two weeks after treatment, the VIM genetic level of uterine tissue in model and transplantation group had significant difference, and the rest two groups were no significant difference compared with control group (Pmodel=0.006, Ptransplantation=0-042, Pestrin=0.079, Pcombined=0.199). Among three treatmental groups, the transplantation group(P=0.116) and estrin group (P=0.095) were no significant difference compared with combined group respectively.④Four weeks after treatment, the VIM genetic level of uterine tissue in combined group had significant difference, and the rest three groups were no significant difference compared with control group (Pmodel=1.000, Ptransplantation=0.136, Pestrin=0.057,Pcombined=0.042). Among three treatmental groups, the transplantation group(P=0.151) and estrin group (P=0.090) were no significant difference compared with combined group respectively.(3) The analysis of ESR1gentic level①Before treatment, the ESR1genetic level of uterine tissue in all experimental groups were significant higher than that in control group (Pmodel=0.000, Ptransplantation=0.000,Pestrin=0.000, Pcombined=0.10). Among three treatmental groups, the transplantation group(P=0.340) and estrin group (P=O.835) were no significant difference compared with combined group respectively.②One week after treatment, the ESR1genetic level of uterine tissue in transplantation and combined group had significant difference, and the rest two groups were no significant difference compared with control group (Pmodel=0.993, Ptransplantation=0.014, Pesrin=0.214, Pcombined=0.024). Among three treatmental groups, the transplantation group(P=0.023) and estrin group (P=0.025) had significant difference compared with combined group respectively.③Two weeks after treatment, the ESR1genetic level of uterine tissue in combined group had significant difference, and the rest three groups were no significant difference compared with control group (Pmodel=0.971, Ptransplantation=1.000, Pestrin=0.078, Pcombined=0.019). Among three treatmental groups, the transplantation group(P=0.124) and estrin group (.P=0.251) were no significant difference compared with combined group respectively.④Four weeks after treatment, the ESR1genetic level of uterine tissue in combined group had significant difference, and the rest three groups were no significant difference compared with control group (Pmodel=0.888, Ptransplamation=0.544, Pestrin=0.112, Pcombined=0.002). Among three treatmental groups, the transplantation group(P=0.136) and estrin group (P=0.463) were no significant difference compared with combined group respectively.(4) The analysis of TGF-B1gentic level①Before treatment, the TGF-B1genetic level of uterine tissue in all experimental groups were significant higher than that in control group (Pmodel=0.000, Ptransplantation=0.000,Pestrin=0.000, Pcombined=0.000). Among three treatmental groups, the transplantation group(P=0.467) and estrin group (P=0.987) were no significant difference compared with combined group respectively.②One week after treatment, the TGF-B1genetic level of uterine tissue in estrin group was no significant difference, and the rest three groups had significant difference compared with control group (Pmodel=0.009,Ptransplantation=0.032, Pestrin=0.132,Pcombined=0.014). Among three treatmental groups, the transplantation group(P=0.014) and estrin group (P=0.013) had significant difference compared with combined group respectively.③Two week after treatment, the TGF-B1genetic level of uterine tissue in transplantation group was no significant difference, and the rest three groups had significant difference compared with control group (Pmodel=0.006, Ptransplantation=0.079,Pestrin=0.001,Pcombined=0.000). Among three treatmental groups, the transplantation group(P=0.009) had significant difference and estrin group (P=0.936) was no significant difference compared with combined group respectively.④Four week after treatment, the TGF-B1genetic level of uterine tissue in estrin and combined group was no significant difference, and the rest two groups had significant difference compared with control group (Pmodel=0.214, Ptransplantation=0.830, Pestrin=0.000,Pcombined=0.000). Among three treatmental groups, the transplantation group(P=0.042) had significant difference and estrin group (P=0.058) was no significant difference compared with combined group respectively.6. The results of protein expression tested by Western blot was showed as follows:(1) The analysis of the CK protein quantification of uterine tissue①Before treatment, there was significant difference among all groups (F=19.402,.P=0.002). The experimental groups had significant difference compared With control group (Pmodel=0.000, Ptransplantation=0.000, Pestrin=0.000, Pcombined=:0.000). Among three treatmental groups, the transplantation group(P=0.755) and estrin group (P=0.646) were no significant difference compared with combined group respectively.②One week after treatment, there was significant difference among all groups (F=1264.341, P=0.000). The experimental groups had significant difference compared with control group (Pmodel=0-000, Ptransplantation=0.000,Pestrin=0.000, Pcombined=0.000). Among three treatmental groups, the transplantation group(P=0.851) and estrin group (P=0.329) were no significant difference compared with combined group respectively.③Two weeks after treatment, there was significant difference among all groups (F=2038.144, P=0.000). The experimental groups had significant difference compared with control group (Pmodel=0-000, Ptransplantation=0.000, Pestrin=0.000, Pcombined=0.000). Among three treatmental groups, the transplantation group(P=0.002) and estrin group (P=0.002) had significant difference compared with combined group respectively.④Four weeks after treatment, there was significant difference among all groups (F=105.979, P=0.000). The experimental groups had significant difference compared with control group (Pmodel=0.000, Ptransplantation=0.000, Pestrin=0.000, Pcombined=0.000). Among three treatmental groups, the transplantation group(P=0.300) was no significant difference and estrin group (P=0.001) had significant difference compared with combined group respectively.(2) The analysis of the ESR1protein quantification of uterine tissue①Before treatment, there was significant difference among all groups (F=1637.295, P=0.000). The experimental groups had significant difference compared with control group (Pmodel=0.000,Ptransplantation=0.000, Pestrin=0.000, Pcombined=0.000). Among three treatmental groups, the transplantation group(P=0.616) and estrin group (P=0.350) were no significant difference compared with combined group respectively.②One week after treatment, there was significant difference among all groups (F=1781.038, P=0.000). The experimental groups had significant difference compared With control group (Pmoder=0.000, Ptransplantation=0.000, Pestrin=0.000, Pcombined=0.002). Among three treatmental groups, the transplantation group(P=0.000) and estrin group (P=0.000) had significant difference compared with combined group respectively.③Two week after treatment, there was significant difference among all groups (F=865.779, P=0.000). The experimental groups had significant difference compared with control group (Pmodel=0.000, Ptransplantation=0.000, Pestrin=0.000, Pcombined=0.043). Among three treatmental groups, the transplantation group(P=0.000) and estrin group (P=0.000) had significant difference compared with combined group respectively. ④Four weeks after treatment, there was significant difference among all groups (F=1781.038, P=0.000). The ESR1protein quantification in model, BMSCs transplantation and estradiol treatment groups were significantly higher than that in cintrol group, and the combined group was almost alike control group CPmodei=0.000, Pransplantation=0.000, Pestrin=0.000, Pcombined=0.248). Among three treatmental groups, the transplantation group(P=0.000) and estrin group (P=0.000) had significant difference compared wi... |
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