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The incidence of spontaneous abortion （SAB） has been increasing year by year， and its etiology is complex， with limited treatment options， which poses a serious threat to social stability. The "disease-syndrome-therapy" research model can significantly improve the clinical efficacy of traditional Chinese medicine （TCM） for preventing miscarriage， but there has always been a lack of key and recognized diagnostic and treatment evaluation markers， which need to be further explored to establish a scientific and unified evaluation standard system. It is proposed to collect existing "disease-syndrome-therapy" SAB animal models， transplant and improve the model evaluation indicators， evaluate the degree of match between SAB animal models and the clinical characteristics of TCM and Western medicine diseases and syndromes， and compare the advantages and disadvantages of different SAB animal models in terms of construction methods， target selection， and evaluation indicators. In addition， the frontiers of TCM experimental research will be explored. In view of the current status and related bottlenecks of molecular biomarkers research on SAB TCM animal models， a single-cell multimodal omics research strategy will be proposed to break through the related evaluation defects of the "disease-syndrome-therapy" SAB and analyze the differences in various cell types， cell subpopulations， spatiotemporal trajectories， and gene expression in the mother-fetal interface tissue at the single-cell level. This will provide accurate guidance and model animal platform support for the in-depth study of disease-syndrome models， Zang-fu biology， and novel targeted drugs. It will also provide a basis for establishing a stable and repeatable "disease-syndrome-therapy" SAB animal model and evaluation indicator system， which is beneficial for the long-term development of TCM reproductive animal model research.
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ObjectiveTo explore difference in the mechanism of Shoutaiwan， a representative kidney-tonifying and abortion-preventing formula， and Juyuanjian， a typical spleen-invigorating and abortion-preventing formula in reversing the pathology of decidua of spontaneous abortion （SA） patients and to expound the connotation of "uterine collaterals connecting kidney" and "fetal collaterals connecting spleen" theory. MethodThe targets of SA were retrieved from GeneCards， followed by gene ontology-biological process （GO-BP） annotation. Based on Cytoscape and previous research， the main processes and core targets were screened out. High-performance liquid chromatography-mass spectrometry （HPLC-MS） was used to identify the potential active components of Shoutaiwan and Juyuanjian and the regulatory networks were constructed. SA was induced in rats and the model rats were treated with Shoutaiwan and Juyuanjian at the same unit. Hematoxylin and eosin （HE） staining， transmission electron microscopy （TEM）， enzyme-linked immunosorbent assay （ELISA）， immunohistochemistry （IHC）， immunofluorescence （IF）， and other methods were employed to verify the mechanisms against miscarriage. ResultThe dysregulation of cell adhesion， inflammatory response， cell death， and angiogenesis was the core pathological process of SA. A total of 13 potential specific active components of Shoutaiwan and 14 active components of Juyuanjian were screened out. The regulatory networks showed that the potential active components of the two prescriptions modulated vascular endothelial growth factor （VEGF）， interleukin （IL）-2， estrogen receptor （ESR）-1， matrix metalloproteinase-9 （MMP-9）， and other targets to regulate the pathological process of SA. The two can significantly improve the pregnancy rate and the integrity rate and blood supply of decidua cells， control the apoptosis morphology and the expression of estrogen （E2）， progesterone （P）， and its receptor， and down-regulate the expression of MMP-2， MMP-9， IL-2， and IL-6 in decidua tissue of SA rats. At the same time， they up-regulated the expression of anti-apoptotic protein B-cell lymphoma-2 （Bcl-2） and IL-4. Shoutaiwan significantly up-regulated the expression of VEGF， and Juyuanjian significantly down-regulated the expression of E-cadherin （E-Cad）. ConclusionBoth Shoutaiwan and Juyuanjian regulate the core pathological process of SA to prevent miscarriage. At the same unit， Shoutaiwan is overall superior to Juyuanjian. Shoutaiwan is better than Juyuanjian in regulating angiogenesis and Juyuanjian is superior to Shoutaiwan in regulating cell adhesion. This conclusion can partly explain the biological basis of "treating the same disease with different methods"， and provide objective data reference for the identification of quality marker （Q-marker） of anti-miscarriage Chinese medicine and further study of formula-syndrome metabolome.
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This study aimed to analyze the effect of the external method of tonifying kidney and promoting blood circulation on endometrial morphology of rats with kidney deficiency and blood stasis model. A total of 50 normal healthy unmated female SD rats of proestrus were selected with vaginal smear. Rats were randomly divided into the blank group, model group, high dose group and low dose group with the prescription of tonifying kidney and promoting blood circulation and theWu-Zi (WZ) group. Except the blank group, intragastric administration of hydroxyurea was given to other group to establish the kidney deficiency model. Meanwhile, clyster of distilled water was given to the blank group and the model group. And clyster of high dose and low dose prescription of tonifying kidney and promoting blood circulation was given to the high dose and low dose group, respectively. The intragastric administrations ofWu-Zi Yan-Zong(WZYZ) pills were given to theWu-Zi (WZ) group. On the 4th day of pregnancy, 10% of the macromolecule dextran was quickly injected to the caudal vein to induce blood stasis model 1 hour after the last medication administration. The uterus tissue section was observed by HE dyeing. The results showed that the thickness of covering epithelium of endometrium in the model group was lower than the blank group (P < 0.05). There were significant differences on the thickness of covering epithelium of endometrium in the high dose group and low dose group with the prescription of tonifying kidney and promoting blood circulation, the WZ group and the model group (P < 0.01). The sum of gland number and gland area as well as the maximum diameter / minimum diameter of the model group was significantly lower than that of the blank group (P< 0.05, orP < 0.01). The shape factor was the closest to “1”, which had significant difference compared with the blank group (P < 0.01). Compared with the model group, the sum of gland number and glandular lumen of the high dose and low dose group were increased at different levels (P < 0.05, orP < 0.01). The sum of gland number, glandular lumen area, shape factor, and maximum diameter / minimum diameter of WZ group had significant difference compared with the model group (P < 0.05, orP < 0.01). The sum of interstitial cells’ nucleus area, integral optical density of interstitial cells’ nucleus and number of interstitial blood vessels in the high dose and low dose group were significantly higher compared to the model group (P < 0.05, orP < 0.01). The sum of interstitial cells’ nucleus area and the integral optical density of interstitial cells’ nucleus in the WZ group were obviously higher than the model group (P < 0.01). It was concluded that the external method of tonifying kidney and promoting blood circulation can effectively improve the endometrial morphology of kidney-deficiency and blood-stasis rat model, promote the synchronous development of endometrial gland and stroma, in order to play a role to improve the endometrial receptivity.
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This study was aimed to establish and evaluate obese polycystic ovary syndrome (PCOS) rat model. Rats were randomly divided into the blank control group, high-fat model group, insulin (INS) combined human chorionic gonadotropin (HCG) model group, and INS combined HCG plus high fat emulsion model group. The obese PCOS rat model was induced by subcutaneous injection of INS and HCG on the nape, respectively. The intragastric administration of high fat emulsion was also used in the PCOS rat model establishment. The estrous cycle of rat was monitored. The detection was also made the weight increasing rate of rats, luteinizing hormone (LH), follicle-stimulating hormone (FSH), estradiol (E2), testosterone (T), high-density lipoprotein (HDL), low density lipoprotein (LDL), triglyceride (TG), cholesterol (TC), fasting plasma glucose (FPG), fasting insulin (FINS), 2 h blood glucose and 2 h INS. Calculation was made on the HOMA index, related viscera index and bilateral ovaries HE staining. The results showed that when INS combined HCG improved method (INS combined HCG plus high fat emulsion model group) rats were sacrificed, the body weight, weight increasing rate, ovary viscera index, T, LH, and HOMA index were significantly higher than that of the blank control group, high-fat model group and INS combined HCG model group (P < 0.05,P < 0.01); HDL was significantly decreased (P < 0.05,P < 0.01). FPG, FINS, and 2 h INS of the INS combined HCG improved method group were significantly higher than that of the high-fat model group (P <0.01). It was concluded that INS combined HCG improved method was one of the ideal animal model establishment methods in the pathogenesis study of PCOS.
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By combing differe nt modelin g factors and evaluation indicators of experimental animals of polycystic o-vary syndrome (PCOS), application scope, advantages and disadvantages were analyzed among various kinds of animal models, in order to screen models of etiology and pathogenesis in traditional Chinese medicine (TCM). Through the collection of 77 Chinese literatures published since 1979, the database on PCOS of modeling factors (i.e., animal species, age, modeling reagent and dose, etc.) and evaluation indicators (i.e., ovarian morphology and hormone levels) were established, analyzed and summarized. The results showed that the experimental PCOS models established by researchers both at home and abroad included androgen method, combination method of androgen with HCG, combi-nation method of progestrone with HCG, estrogen method, aromatic cyclase inhibitor method, combination method of HCG with insulin, combination method of androgen with insulin, insulin method, and etc. The SD and Wistar rats at the age of 3-50 days were commonly used. Chemicals, such as dehydroepiandrosterone (DHEA), testosterone propi-onate, sodium prasterone sulfate, dihydrotestosterone (DHT), levonorgestrel, estradiol valerate, letrozole, insulin and HCG were chosen to build the animal models. The evaluation indicators included ovarian morphology, the total num-ber of follicles at different levels, sex hormones (E2, P, T, FSH, LH) and insulin resistance (IR) index (FINS, FPG, HOMA). It was concluded that each modeling method described above had its own advantages and disadvantages, which cannot completely simulate clinical onset process of PCOS patients. Researchers should choose appropriate reagent and evaluation index according to different purpose. DHEA, DHT slow-release, progesterone combined with HCG modeling methods are more suitable for the study of the role of IR in the pathogenesis of PCOS. Kidney-defi-ciency and dampness-phlegm is fundamental etiology and pathogenesis of obese type of PCOS. DHEA method, com-bination method of levonorgestrel with HCG, aromatic cyclase inhibitor method conformed obese type of PCOS. The evaluation indicators included body weight, sex hormones, IR index and ovarian morphology. The method of kidney-tonifying and phlegm-reducing was often used in the treatment of obese type of PCOS in the clinical practice.
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Objective To explore the current situation of the women of childbearing age who suffered from kidney deficiency,as well as the distribution regularities of the different syndromes of kidney deficiency,so as to guide the clinical application and scientific research.Methods 1 343 women at the areas including communities of Wangjiang,Changshouyuan and Qintai road in Chengdu,and the towns of Chongzhou,Xinfan and Shibantan were investigated from November 4th,2006 to May 3rd,2007.The results was input with Epidata,and tested with ? 2 test of SPSS13.0.Result There were 925 people suffered from kidney deficiency,including 744 cases of kidney yin deficiency,572 cases of kidney yang deficiency,467 cases of kidney qi deficiency and 357 cases of kidney essence deficiency,with statistical significant difference(P