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Objective:This study explores the key core targets of Guishenwan in the treatment of thin endometrium and related signaling pathways through the method of network pharmacology，and further uses animal experiments to verify the obtained targets and verify that Guishenwan are effective for thin endometrium. Method:Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform（TCMSP） was used to retrieve the effective chemical components，active component targets and target abbreviations of the eight Chinese medicines in Guishenwan，the GeneCards database and Online Mendelian Inheritance in Man（OMIM）database were used to retrieve thin endometrial related targets gene.Use Wayne software to take the intersection of the drug target of Guishenwan and the disease target of the thin endometrium，and import the intersection target into the STRING database and Cytoscape 3.7.2 software for visual analysis to obtain the "drug-disease" protein protein interaction（PPI） network， then input the intersection target into Enrichr database and DAVID database for gene ontology（GO） enrichment analysis and Kyoto encyclopedia of genes and genomes（KEGG） enrichment analysis. Using the obtained possible core and key targets as the theoretical basis，a thin endometrial model in rats was established. After Guishenwan and estrogen intervention for 21 days，the endometrial thickness of rats was observed by hematoxylin-eosin staining（HE） staining. Western blot and quantitative real time polymerase chain reaction（Real-time PCR） detect the protein and mRNA expression levels of the four core key targets of epidermal growth factor receptor（EGFR），matrix metalloproteinase 9（MMP9），interleukin-1beita（IL-1<italic>β</italic>） and mitogen activated protein kinase 14（MAPK14）. Result:The Venn software obtained 130 intersection targets in total， imported 130 intersection targets into the STRING database and Cytoscape database，and obtained a protein interaction network diagram including 33 nodes and 107 edges. DAVID 6.8 database for GO analysis. The function annotation analysis involving 167 biological processes（BP），22 cell components（CC），39 molecular functions（MF）. DAVID 6.8 database for KEGG enrichment analysis， and thin endometrium related A total of 34 pathways. Western blot and Real-time PCR were used to analyze the expression results of EGFR，MMP9，IL-1<italic>β</italic>，MAPK14 protein and genes in the endometrial tissues of the 6 groups of rats. Guishenwan can enhance the expression of EGFR，MMP9，IL-1<italic>β</italic>，MAPK14 protein and mRNA on the thin endometrium. Conclusion:According to the theoretical analysis of network pharmacology and the results of animal experiments，it is found that Guishenwan can effectively improve the related indicators of thin endometrium，and promote the expression of EGFR，MMP9，IL-1<italic>β</italic>，MAPK14 protein and genes. The intimal tissue proliferates and improves the symptoms of thin intima.
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Objective:To explore the expression of abnormal spindle-like microcephaly-associated (ASPM) in liver cancer tissues and clarify its prognostic relationship with clinicopathological features of liver cancer after liver transplantation.Methods:Immunohistochemistry was employed for detecting the expression of ASPM in 72 liver cancer tissues and 36 adjacent tissues of liver cancer liver transplant recipients fulfilling the Hangzhou criterion. In conjunctions with clinicopathological data, the correlation between the expression level of ASPM in liver cancer tissues and the clinicopathological characteristics and the post-transplantation prognosis for liver cancer were statistically analyzed.Results:During a median follow-up period of 29 months, 20 patients relapsed and 8 died after transplantation. Immunohistochemical results indicated that the high-expression rates of ASPM were 58.3% and 25.0% in liver cancer and adjacent tissues ( P=0.001). The difference was statistically significant. The high-expression rate of ASPM was significantly higher in liver cancer tissues than that in adjacent tissues. The expression level of ASPM was not correlated with gender, age, smoking/alcoholic history, hepatitis history, preoperative level of alpha-fetoprotein (AFP), tumor size, tumor load or vascular tumor thrombus ( P>0.05). And the postoperative high-expression rates of ASPM were 51.0% and 76.2% in pathological differentiation type Ⅰ-Ⅱ and Ⅲ-Ⅳ groups ( P=0.049). The difference was statistically significant. The wrose pathological differentiation type of liver cancer, the higher expression level of ASPM in liver cancer tissue. In liver cancer tissues, the overall 1/3/5-year survival rates of ASPM high/low-expression group were 97.6%, 80.6%, 80.6% and 93.3%, 89.7% and 89.7% respectively ( P>0.05). There was no statistical significance. And 1/3/5-year long-term disease-free survival rates were 78.6%, 55.5%, 55.5% and 86.3%, 86.3% and 86.3% respectively ( P=0.036). The difference was statistically significant. The disease-free survival rate was lower in ASPM high-expression group and post-transplantation prognosis was worse. Conclusions:The expression of ASPM is significantly higher in liver cancer tissues than that in adjacent tissues. And the expression level of ASPM in liver cancer tissues is correlated with pathological differentiation types of liver cancer and has an impact on tumor-free survival of patients after liver transplantation for liver cancer.
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Objective To evaluate the efficacy of in-situ split liver transplantation (ISSLT) in children.Methods From June 2015 to August 2018,10 liver grafts from DBD were split in-situ.All the donors were male,and the median age of the donors was 28.5 year old (18-48 year).One left half graft and 9 left lateral lobe grafts (including 2 reduced size grafts) were transplanted to 10 pediatric recipients.Four grafts were transplanted in our center,and the rest 6 grafts were shared to other two transplant center.The primary diseases of the recipients included biliary atresia (8/10),hepatic sinus obstruction syndrome (1/10) and Alagille syndrome (1/10).The median age of the recipients was 10 month (7 month-11 year),and the mean body weight was 9.8 ± 6.6 kg (5-28 kg).Results All liver grafts were split in-situ.The mean split time of liver grafts was 88.5 ± 18.9 min.The mean weight of split grafts was 336.7-± 85.4 g.All recipients were subjected to piggyback liver transplantation.Operation time was 542.5 ± 112.1 min.Anhepatic time was 52.0 ±-13.5 min.GRWR was (3.98 ±0.96)％.GRWR of two cases was more than 5％,so segment Ⅲ was partially reduced.During the follow-up period,9 cases were alive and 1 case died due to multiple organ failure 1 day after liver transplantation.Conclusions ISSLT can enlarge the graft pool for children and achieve good results.
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<p><b>OBJECTIVE</b>To study the role of estrogen (E2), estrogen receptor (ER) and aromatase (P450arom) in the pathogenesis of uterine adenomyosis.</p><p><b>METHODS</b>Paraffin-embedded specimens of the uterine tissue from patients with uterine adenomyosis and patients with cervical lesions (CIN; control) were examined for expressions of E2, ER and P450arom by immunohistochemistry and ELISA. The cells isolated from the lesions of patients with adenomyosis were cultured in vitro, and the changes in cell growth in response to treatments with E2, ER inhibitor, ER inhibitor + E2, estrogen deprivation, and estrogen deprivation+ ICI182780 were assessed using CCK-8 method.</p><p><b>RESULTS</b>The expression levels of E2, ER, and P450arom were significantly higher in adenomyosis ectopic lesions and eutopic endometrium than in the myometrium and endometrium in the control group (P<0.05); no significant difference in E2 and P450arom expressions was found between adenomyosis ectopic lesions and eutopic endometrium (P>0.05), while the expression levels of ER in ectopic lesions was significantly higher than that in eutopic endometrium. The cell inhibition rates were similar between ER inhibitor group and ER inhibitor + Estrogen activation group (P>0.05), and was significantly higher in estrogen deprivation+ ER inhibitor group than in estrogen deprivation group (P<0.05).</p><p><b>CONCLUSION</b>The high expression levels of E2, ER, and P450arom in adenomyosis ectopic lesions and eutopic endometrium promote uterine adenomyosis cell proliferation, in which process E2 combines with ER to execute its biological effect; ER also promotes the occurrence and development of uterine adenomyosis through other pathways.</p>