Objective To observe the effect of Juli Sanjie Pill(JSP) on estrogen receptor(ER)? and ER? expression in the myometrium and hysteromyoma tissue,and to explore the correlation of ER? and ER? expression levels with the incidence of hysteromyoma. Methods Forty hysteromyoma patients were divided into two groups: 20 patients with operative indications after oral use of JSP and asking for operation,were enrolled in the treatment group,and other 20 patiens without mediation of medicine but asking for operation were in the control group.The ER? and ER? expression levels in hysteromyoma tissue and the surrounding normal myometrium of the two groups were detected by radioimmunoassay.Results There presented the expression of ER? and ER? in the hysteromyoma tissue and the surrounding normal myometrium of the two groups,and the levels in the hysteromyoma tissue were higher than that in the myometrium(P0.05).Conclusion The incidence of hysteromyoma is correlated with the local expression of ER? and ER? in the uterus.The therapeutic mechanism of JSP for hysteromyoma is probably related with the decrease of ER? and ER? expression levels in hysteromyoma tissue and with the decrease of ER? level in the surrounding normal myometrium.

Objective The purpose of this study was to compare the adenomyosis models in nude mice generated by four different methods,and to find out an optimal modeling method, and to provide an ideal animal model for exploring pathogenesis and experimental treatment of uterine adenomyosis. Methods 1. 80 female healthy nude mouse were divided randomly into 4 groups: Intraperitoneal implantation group, subcutaneous implantation group, intraperitoneal injection group, and subcutaneous injection group. The transplants were taken for pathological examination at 4 weeks after surgery. Results The success rate of intraperitoneal implantation group was 95%,and that of the subcutaneous implantation group was 45%,while the success rate of intraperitoneal injection group and subcutaneous injection group was 0%. Conclusions Establishment of a nude mouse model of uterine adenomyosis by intraperitoneal implantation method has a high success rate and with good stability, and is an ideal mouse model of human-derived uterine adenomyosis.

Thin endometrium is an important factor influencing the in vitro fertilization and embryo transfer, and there is a lack of effective therapy in the treatment of thin endometrium.The aim of this study was to explore a stable animal model of effective thin endometrium, and to promote the research on thin endometrium pathogenesis and provide experimen-tal basis of treatment.To analyze a variety of establishments of endometrial damage animal model reported in the domestic and foreign literatures,it is concluded that perfusing 95% ethanol into uterine cavities of rats can establish a rat model of thin endometrium,and put forward some experience and methods for its improvement.

Objective To observe the changes in estrous cycle and vaginal smears in ovarectomized NOD/SCID mice.Methods To continuously observe the estrous cycle time by vaginal smears of NOD/SCID mice in consecutive nine days, twice daily.After ovariectomy, the changes of estrous cycle were observed by vaginal smears for 7 days.Results The estrous cycle in NOD/SCID mice was 4-6 days.Regular estrous mice accounted for 80%.There was no significant correlation between vaginal opening and estrous cycle status.After ovariectomy, the vaginal smears showed characteristics of metestrus or diestrus.Conclusions Vaginal smear cytology can be used to determine the estrous cycle and characteris-tics of NOD/SCID female mice.The ovariectomized operation of NOD/SCID female mice is effective.

BACKGROUND: Thin endometrial diseases are a challenge in clinical treatment at present. Scholars have found that bone marrow mesenchymal stem cells (BMSCs) transplantation has its unique curative effect and advantages, but few studies have been conducted on pathway or gene control. OBJECTIVE: To observe the effect of BMSCs transplantation in rats with thin endometrium based on the HOXA10 regulatory network. METHODS: Twenty-one adult female Sprague-Dawley rats of SPF grade (provided by the Animal Experimental Center, Guangzhou University of Chinese Medicine in China) were randomly divided into three groups (n=7/group): control group, model group, and BMSCs group. In the latter two groups, a thin endometrium model was prepared in each rat by filling the uterine cavity with 95％ ethanol. In the control group, normal saline was injected to fill the uterine cavity of rats. After extraction of ethanol or normal saline, the rats in the BMSCs group were injected intrauterinely with 1 mL of BMSCs suspension (1×1010 cells/L) , and those in the control and model groups were given the same volume of normal saline. After two estrous cycles, the uterus of each rat was removed. Hematoxylin-eosin staining was used to measure the thickness of the endometrium. Immunohistochemistry was used to detect the expression of vimentin, keratin, vascular endothelial growth factor, leukemia inhibitory factor and integrin αvβ3. qRT-PCR was used to detect the relative transcription of HOXA10 and miR-196 b. RESULTS AND CONCLUSION: (1) Compared with the control group, the endometrial thickness of the rats were significantly thinner in the model and BMSCs groups (P < 0.05) , while the endometrial thickness in the BMSCs group was thicker than that in the model group (P < 0.05). (2) The mean absorbance values of endometrial vimentin, keratin, vascular endothelial growth factor, leukemia inhibitory factor and integrin αvβ3 were highest in the control group, higher in the BMSCs group and lowest in the model group, and there were significant differences between groups (P < 0.05). (3) The relative transcript level of HOXA10 gene in the model and BMSCs group was significantly lower than that in the control group, while the relative transcript level of HOXA10 gene in the BMSCs group was significantly higher than that in the model group (P < 0.05). The relative transcript level of miR-196 b in the model and BMSCs groups was significantly higher than that in the control group (P < 0.05) , while the relative transcript level of miR-196 b in the BMSCs group was lower than that in the model group (P < 0.05). (4) HOXA10 was negatively correlated with miR-196 b gene, HOXA10 was positively correlated with the protein expression to different extents, and miR-196 b gene was negatively correlated with the protein expression to different extents. These findings suggest that BMSCs transplantation can improve the endometrial thickness and relevant protein levels of thin endometrium rats to some extent, which may be attributed to the negative regulation of HOXA10 gene by miR-196 b, and HOXA10 gene further promotes the expression of vascular endothelial growth factor, leukemia inhibitory factor and integrin αvβ3 proteins.

Objective To establish a rat xenograft model of human uterine leiomyoma using immunosuppressive a-gent and provide a useful tool for the study on uterine leiomyoma. Methods Intragastric administration with immunosup-pressive agent mycophenolate mofetil(MMF)(40 mg/kg)was given to rats for two weeks before the surgery until the end of the experiment. 20 SPF female Wistar rats were randomly divided into 4 groups after abdominal transplantation of human leiomyoma tissues:group A received femoston containing 0.4 mg/kg estradiol and 2 mg/kg dydrogesterone, group B re-ceived estadiol 0.4 mg/kg,group C received dydrogesterone 2 mg/kg,and group D served as the control group, received distilled water 1 mL/200 g. All rat received the corresponding drugs once per day for 2 days. Samples were taken at 4 weeks after the surgery to observe the pathology of the tumor tissues. Results The modeling success rates were 90% in the group A,40 % in the group B,and 0% in the groups C and D. Conclusions Rat xenograft model of human leiomyoma can be successfully established using an immunosuppressive agent femostone with a high modeling success rate and low cost. It can be used as a new animal model for the study of transplanted leiomyoma.

Objective To investigate the levels and significance of Th17 cells and regulatory T cells (Treg) in peripheral blood of children with allergic rhinitis during pollen and non-pollen seasons.Methods Thirteen children with hay fever, 10 children with house dust mite(HDM)-allergic asthma and 10 healthy children were recruited into this study.Percentages of Th17 and Treg cells were detected by flow cytometry.Levels of IL-17, IL-10 and TGF-β in cell culture supernatants were measured by ELISA.Results (1) The percentages of Th17 cells in children with allergic rhinitis [(3.4±2.4)%] were significantly higher than those in HDM-allergic asthmatics [(2.1±1.6)%] and those in healthy children [(0.5±0.3)%] during pollen season (both P<0.05).The levels of Treg cells in allergic rhinitis group [(2.1±1.3)%] and in HDM-allergic asthma group [(3.6±1.9)%] were significantly lower than those in healthy control group [(5.5±2.8)%] (both P<0.05).The levels of Th17 cells [(3.0±1.9)% vs (3.4±2.4)%, P<0.05] and ratios of Th17/Treg cells [(1.4±1.0)% vs (1.7±1.5)%, P<0.05] in children with allergic rhinitis were significantly decreased during non-pollen season as compared with those during pollen season, but the levels of Treg cells were up-regulated [(2.4±1.6)% vs (2.1±1.3)%, P<0.05].(2) Correlation analysis revealed that the ratios of Th17/Treg cells were positively correlated with the concentrations of FeNO (fractional concentration of exhaled NO) (r=0.321, P<0.05) and the counts of circulating eosinophils (r=0.198, P<0.05) in children with allergic rhinitis during pollen season.Conclusion The imbalanced Th17 and Treg cells in children with allergic rhinitis during pollen season might play a vital role in the regulation of allergic airway inflammation.

Objective To investigate the changes in percentage and function of CD4+CD25+regu-latory T cells ( Tregs) in peripheral blood of patients with hay fever. Methods A total of 20 patients with hay fever, 20 patients with house dust mite-induced allergic asthma and 20 healthy subjects were enrolled in this study. Peripheral blood samples were collected from all subjects to isolate PBMCs. Percentages of Tregs in PBMCs were measured by flow cytometry. CD4+CD25+ Tregs and CD4+CD25-T cells ( Teffs) were isola-ted by immunomagnetic cell sorting. Effects of CD4+CD25+Tregs on the proliferation of Teffs were evaluated by MTT assay. Expression of Foxp3 and TGF-β1 at mRNA level was analyzed by RT-PCR. Results During the pollen season, the percentage of circulating Tregs in patients with hay fever [(1. 82+0. 82)％] was sig-nificantly lower than that in patients with house dust mite-induced allergic asthma [(2. 96±1. 34)％] and health subjects [(5. 78±2. 29)％] (both P<0. 05). Expression of Foxp3 at mRNA level was significantly reduced in patients with hay fever (0. 46±0. 25) as compared with that of the house dust mite-induced aller-gic asthma (0. 64±0. 31) and healthy control (1. 04±0. 21) groups (both P<0. 05). Expression of TGF-β1 at mRNA level in both hay fever (0. 34±0. 27) and house dust mite-induced allergic asthma (0. 43±0. 31) groups was lower than that of the healthy control group (0. 99±0. 34). Treg-mediated suppression of Teff proliferation was significantly decreased in patients with hay fever [(17. 1±8. 4)％] as compared with that in patients with house dust mite-induced allergic asthma [(21. 4±9. 1)％]) and healthy subjects [(36. 0± 13. 9)％] (P<0. 05). Conclusion Decreased percentage and defective function of Tregs might be one of the major causes for the occurrence and development of hay fever in children during the pollen season.

Objective: To analyze the dynamic changes in the expression and function of peripheral type Ⅱ innate lymphoid cell (ILC2) subpopulation and the activity of signal transducers and activators of transcription (STAT6) in children with hay fever during pollen season. Methods: A total of 10 patients with hay fever, 10 patients with house dust mite (HDM)-sensitized asthma and 12 healthy controls (HC) were enrolled in this study. Changes in peripheral ILC2 and the intracellular expression of Th2-related cytokines were detected by flow cytometry during and outside the pollen season. Peripheral Lin^- cell population was isolated from each group and cultured with the presence of IL-25 or IL-33 for 7 d. The concentrations of IL-5 and IL-13 in culture supernatants were measured by ELISA. Expression of phospho-STAT6 at protein level was quantified by Western blot. Results: Within the pollen season, the percentage of peripheral ILC2 cells was significantly higher in children with hay fever [(23.09±7.86)%] than in children with HDM-sensitized asthma [(6.84±3.85)%, P<0.05] and healthy children[(1.69±0.87)%, P<0.05]. In the non-pollen season, the peripheral ILC2 cells in children with hay fever presented a decreasing trend [(11.30±2.45)%], but was still higher than that in HDM-sensitized asthmatics [(3.76±1.96)%, P<0.05] and HC [(1.32±0.91)%, P<0.05] at the same time point. Moreover, peripheral IL-13⁺ ILC2 cells in children with hay fever [(6.94±3.16)% vs(4.17±1.98)%, P<0.05] and in HDM-sensitized asthmatics [(1.89±0.70)% vs(1.44±0.55)%, P<0.05] during the pollen season were significantly higher than those in the non-pollen season. After the in vitro stimulation with IL25 or IL-33, the levels of IL-5 and IL-13 in culture supernatants were both increased in children with hay fever and HDM-sensitized asthmatics, and a synergistic action was observed when IL25 and IL-33 were used in combination. Meanwhile, the protein level of phospho-STAT4 in Lin^- cells was significantly up-regulated in the hay fever group after stimulation with IL25 and IL-33. Conclusions During the pollen season, the abnormal number and function of ILC2 subpopulation in children with hay fever might be another cause of the occurrence of clinical symptoms in a short period of time or acute exacerbation.

Objective To investigate the deep mechanism of Jiawei Shaoyao Gancao Tang (Modified Peony and Licorice Root Decoction) and its medicated serum in treatment of adenomyosis.Methods The cells in ovarian focus tissue were given in vitro primary culture in 6 patients with adenomyosis undergone hysterectomy,and the focus cells were identified by using immunoeytochemistic method.There were 6 groups in this study including high-dose Jiawei Shaoyao Gancao Tang group (high-dose JSGT group,20 mg/mL),low-dose Jiawei Shaoyao Gancao Tang group (low-dose JSGT group,10 mg/mL),Jiawei Shaoyao Gancao Tang medicated serum group (medicated serum group),blank serum group,mifepristone group and blank control group.The focus cells were intervened for 24 h and 48 h respectively in all groups,and then the expression of estrogen (E2) was detected by using enzyme-linked immunosorbent assay (ELISA).The expressions of estrogen receptor (ER) and aromatase P450 (P450arom) were detected by using immunocytochemistic method in all groups.Results The identification results of immunocytochemistic method showed that collected from 6 patients and successfully cultured cells were all focus cells of adenomyosis.The level of E2 decreased significantly in medicated serum group compared with blank control group and blank serum group (P ＜ 0.05).The level of ER decreased significantly in high-dose JSGT group,low-dose JSGT group,medicated serum group and mifepristone group after 24 h (P ＜ 0.05).The total efficacy of decreasing ER was inferior in mifepristone group to that in high-dose JSGT group,low-dose JSGT group and medicated serum group (P ＜ 0.05),and efficacy of decreasing ER was superior after 24 h to that after 48 h in high-dose JSGT group (P ＜ 0.05).The level of P450arom decreased significantly in high-dose JSGT group,low-dose JSGT group,medicated serum group and mifepristone group after 24 h compared with blank serum group and blank control group (P ＜ 0.05),and the efficacy decreasing P450arom was better in high-dose JSGT group,low-dose JSGT group and medicated serum group than that in mifepristone group after 24 h (P ＜ 0.05).The efficacy decreasing P450arom was superior after 24 h to that after 48 h in low-dose JSGT group (P ＜ 0.05).Jiawei Shaoyao Gancao Tang's efficacies of decreasing ER and P450arom were better than mifepristone and these efficacies showed timeliness:the efficacies were better after 24 h than that after 48 h.There was no dose dependence.Conclusion Jiawei Shaoyao Gancao Tang can reduce the levels of ER and P450arom,cut off the positive feedback loop of E2 and P450arom and weaken E2 effect,which may be its mechanism in treatment of adenomyosis.