1.Protective effect and mechanism of trophoblast-derived mesenchymal stem cells against liver injury in a mouse model of autoimmune hepatitis
Long XU ; Wanwan SHI ; Jiandan QIAN ; Shiyun KE ; Yusheng HE ; Xiaofang WANG ; Guiqiang WANG
Journal of Clinical Hepatology 2026;42(9):2116-2125
ObjectiveTo investigate the effect of trophoblast-derived mesenchymal stem cell (T-MSC) on concanavalin A (Con A)-induced immune-mediated liver injury in mice, as well as its association with the changes in inflammatory response. MethodsMorphological assessment, flow cytometry, and tri-lineage differentiation assay were used to observe the biological characteristics of T-MSC. In the pilot experiment, mice were divided into PBS group, T-MSC monotherapy group, and Con A groups at a concentration of 10, 15, or 20 mg/kg to evaluate the safety of T-MSC monotherapy on liver function and determine the appropriate dose for modeling. In the formal experiment, mice were divided into healthy control group (HC group), Con A group, and Con A+T-MSC group, with 5 mice in each group. The mice were given injection of Con A (15 mg/kg) via the caudal vein to establish a mouse model of immune-mediated liver injury, while the mice in the T-MSC monotherapy group were given transfusion of 1×106 T-MSCs via the caudal vein at 3 hours after modeling, and samples were collected 12 hours later. The serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH) were measured; liver histopathological changes were observed, and Ishak inflammatory activity score and inflammatory necrosis area were assessed; the serum levels of inflammatory cytokines were measured; transcriptome sequencing, immunofluorescence assay, and Western blot were used to measure the changes in inflammation-related molecules in liver tissue. The Shapiro-Wilk test was used to assess the normality of continuous data, and the homogeneity of variance test was also performed; a one-way analysis of variance was used for comparison of normally distributed data with homogeneity of variance between multiple groups, and the Tukey’s post-hoc test was used for further comparison between two groups; the Kruskal-Wallis H test was used for comparison of non-normally distributed data with heterogeneity of variance between multiple groups, and the Wilcoxon test was used for further comparison between two groups. ResultsT-MSC had a long spindle shape, showed the characteristics of fibroblast-like morphology and adherent growth, and possessed the osteogenic, chondrogenic, and adipogenic differentiation abilities. Flow cytometry showed high expression levels of CD90, CD105, and CD73 in T-MSC, with a positive rate of 96.8%, 99.0%, and 99.9%, respectively, while there was low or no expression of CD31, human leukocyte antigen DR, CD45, and CD34, with a positive rate of 0.120%, 0, 0.061%, and 0, respectively. Administration of T-MSC alone did not cause obvious abnormalities in ALT, AST, or LDH. Con A at the concentrations of 10, 15, and 20 mg/kg all increased the serum levels of biochemical parameters for liver injury in a dose-dependent manner, with mortality observed in the 20 mg/kg group. In the formal experiment, compared with the HC group, the Con A group had significant increases in the serum levels of ALT, AST, and LDH (P<0.01), and after T-MSC treatment, there were significant reductions in these markers compared with the Con A group (P<0.01). HE staining showed that the Con A group had disordered structure of the liver, accompanied by marked inflammatory cell infiltration and focal/patchy necrosis, and these pathological changes were alleviated after T-MSC treatment. Compared with the HC group, the Con A group had significant increases in Ishak score and the percentage of inflammatory necrosis area (P<0.001), while there were significant reductions in these indicators after T-MSC treatment compared with the Con A group (P<0.01). ELISA showed that compared with the HC group, the Con A group had significant increases in the serum levels of interleukin-21 (IL-21), interleukin-1β (IL-1β), interleukin-2 (IL-2), interleukin-17A (IL-17A), and tumor necrosis factor-α (TNF-α) (P<0.05), and there were significant reductions in these markers after T-MSC treatment compared with the Con A group (P<0.05), with no significant differences in IL-21, IL-1β, IL-17A, and TNF-α compared with the HC group (P>0.05); although there was a significant reduction in the level of IL-2, the Con A+T-MSC group still had a significantly higher level of IL-2 than the HC group (P=0.001). In contrast, compared with the HC group, the Con A group had a significant increase in IL-10 (P<0.05), with a further increase after T-MSC treatment (P<0.05). The transcriptome analysis showed that T-MSC treatment downregulated the expression of the inflammation- and chemokine-related genes including Ccl2, Cxcl10, Il-21, and Il-6 and inhibited the inflammation-related pathways such as TNF, IL-17, nuclear factor-κB, and JAK-STAT. Immunofluorescence assay showed that compared with the HC group, the Con A group had a significant increase in the expression of IL-21 in the liver (P<0.001), and there was a significant reduction in IL-21 after T-MSC treatment compared with the Con A group (P<0.001); the results of Western blot showed the same trend. ConclusionT-MSC can alleviate Con A-induced immune-mediated liver injury in mice, as evidenced by the improvement in liver biochemical parameters, the alleviation of histopathological damage, and the overall downregulation of inflammation-related molecules. T-MSC exerts marked anti-inflammatory and immunomodulatory effects in this model, and IL-21 shows a significant change in inflammatory response and can be used as a candidate molecule for further investigation.
2.Application of the pathological classification of "CCCG-WT-2016" (2019 revision) for treatment of Wilms tumors
Qiao HE ; TouEn Kenneth CHANG ; Wanwan CHEN ; Jing MA ; Ping SHEN ; Jiefeng CHEN ; Xiaoting JIN ; Zhongde ZHANG ; Minzhi YIN
Chinese Journal of Pathology 2020;49(4):324-328
Objective:To describe our experiences in application of the 2019 revision of "CCCG-WT-2016" for the diagnosis of Wilms tumors.Methods:Ninety-one cases of Wilms tumor diagnosed at Shanghai Children′s Medical Center from January 2015 to December 2018 were collected. All cases were reviewed by two senior pathologists, including one from China and the other from Singapore, according to the 2019 revision of "CCCG-WT-2016."Results:The specimens were obtained by core biopsy ( n=21), primary nephrectomy ( n=41), post-chemotherapy nephrectomy/resection ( n=18), or biopsy/resection of metastatic/relapse/post-chemotherapy metastatic lesion(s) ( n=11). The specimens of core biopsy and primary nephrectomy ( n=62) all had favorable histology.Twelve post-chemotherapy nephrectomy cases were subdivided into three risk groups: low risk ( n=0), intermediate risk ( n=10) and high risk ( n=2). Six post-chemotherapy resection cases were subdivided into 3 risk groups:low risk ( n=0), intermediate risk ( n=5) and high risk ( n=1). The remaining 11 cases were comprised of metastatic, relapse, and post-chemotherapy metastatic lesions. The concordance rate of the two senior pathologists was 100%(91/91). Conclusions:The 2019 revision of "CCCG-WT-2016" is clearly written and easy to use. It can serve as the basis of accurate classification for clinical treatment.
3.In-Vitro Anti-HBV Activity of Bushen Jianpi Formula
Wanwan HE ; Ni LIU ; Huiquan XIAO ; Riyong LUO ; Yutong ZHU
Journal of Guangzhou University of Traditional Chinese Medicine 2001;0(01):-
[Objective]: To study the in-vitro anti-HBV activity of Bushen Jianpi Formula (BJF). [Methods] 2.2.15 cell strain was cultured in vitro to observe the effect of BJF on its secretion of HBsAg and HBeAg. [Results] After the addition of BJF for ten days, median toxic concentration of BJF was 37.80g/L, median inhibitory concentrations of HBsAg and HBeAg were 19.05g/L and 9.55g/L, and the therapeutic index was 1.98and 3.96. [Conclusion] It is suggested that BJF has an in-vitro inhibitory effect on the secretion of HBeAg.

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