1.Parathyroid hormone on the adipogenic potential of rat bone marrow mesenchymal stem cells
Fei GAO ; Zhimin GE ; Zhonghao GUO ; Jing YANG
Chinese Journal of Rheumatology 2015;19(4):249-253,封3
Objective To observe the effect of different concentrations of parathyroid hormone 1-34 on the adipogenic potential of rat bone marrow mesenchymal stem cells (BMSCs).Methods ① Rat bone marrow mesenchymal cells were separated and expanded by adherent culture.The morphology of cells was observed and cell surface markers were examined by flow cytometry.② The multi-lineage differentiation capability of cells was examined by culturing cells under conditions favorable for adipogenic and osteogenic differentiation.③ Taken P3 of BMSCs for test,different concentrations of PTH1-34 (0,10-10,10-9,10-8 mol/L) were used to stimulate BMSCs respectively,14 days later,lipoprotein lipase (LPL) activity were measured by enzyme linked immuno-sorbent assay (ELISA),mRNA expression of alkaline phosphatase (ALP) and PPARγ-2 were measured by realiime polymerase chain reaction (PCR).④ Statistical analysis:data were presented as x±s.All statistical analysis was performed with windows Statistical Praduct and Serice Solutions (SPSS) 13.0.One-way analysis of variance (ANOVA) was applied to determine the difference between groups.Least signisicant difference (LSD) was used to determine the difference between the two randomized groups.Differences were considered significant at a value of P<0.05.Results The cells expressed CD44,CD29 but without expression of CD45.By culturing in adipogenic medium for 3 weeks and in osteogenic medium for 4 weeks respectively,and then identified by oil red O and Alizarin red,the cells were successfully induced to adipocytes and osteogenesis.Expressions of LPL were 11.20±0.16,7.62±0.48,5.84±0.57,5.32±0.52,mRNA expressions of PPARγ-2 were 2.80±0.05,1.36±0.23,0.94-±0.11,0.78±0.04,ALP activity were 0.191 ±0.016,0.333±0.024,0.549±0.025,0.684±0.021 respectively.Compared with the control group,different concentrations of PTH1-34 groups could decrease mRNA expression of LPL and PPARγ-2.ALP activity were increased(P<0.05).Conclusion PTH1-34 inhibits BMSCs of adipogenic differentiation and promotes osteogenic differentiation in a dose-dependent manner.
2.Pathological and clinical effects of preoperative arterial infusion chemotherapy on the breast cancers
Zeping YU ; Chunhua GUO ; Yousheng LI ; Zhonghao CHEN ; Jieshou LI
Journal of Medical Postgraduates 2001;14(2):143-145
Objectives:To investigate the pathological and clinical effect of preoperative arterial infusion chemotherapy on breast cancers.Methods:Twenty patients with breast carcinoma received regional arterial angiography by Seldinger's procedure followed by arterial infusion chemotherapy.Sixteen patients who didn't undergo preoperative chemotherapy were selected as controls.All the operation specimens were analyzed by the same pathologist.Results:Histological analysis of the two groups revealed the following results:① cancer tissue necrosis increased in the arterial chemotherapy group;②karyopyknosis,karyorrhexis coagulation and necrosis of cytoplasm around the vascular vessels as well as interstitial edema were found in the tumor tissue,invasion of inflammatory cells,intimal proliferation thrombus and inflammation of vessels could also be seen.All the changes were much severe in the infusion chemotherapy group than in the controlled group;Conclusions:Histological changes are significant after preoperative arterial infusion chemotherapy for breast carcinoma.
3.Advances in research on anti-fatigue effect of phytochemicals
Yawen WANG ; Zhonghao XIN ; Weina GAO ; Changjiang GUO
Military Medical Sciences 2016;40(12):1009-1012
Fatigue not only affects people′s lives and work, but also causes diseases .Numerous studies have shown that some phytochemicals can promote body energy metabolism and regeneration , improve physical condition , resist fatigue growth and accelerate fatigue alleviation .This paper reviewed the recent research progress , domestic and overseas , in the anti-fatigue effect of phytochemicals .
4.Effect of Knocking Down ski on Migration of Astrocytes in Rats
Xin ZHAO ; Xingwen WANG ; Jiangli KOU ; Zhonghao LI ; Yongqiang GUO ; Yamin WU ; Haihong ZHANG
Chinese Journal of Rehabilitation Theory and Practice 2017;23(8):905-911
Objective To investigate the effect of ski gene in migration process of astrocytes in rats. Methods Astrocytes were obtained from rats' cerebral cortex and cultured in vitro. siRNA targeting ski gene and negative control sequences were prepared. The ski-siRNA group, siRNA negative control group and untreated group were set in this experiment. The specific siRNA targeting ski gene was transfected into astrocytes with Lipofectamine?RNAiMAX Reagent. Then the ski protein levels were determined with Western blotting. After transfec-tion, the changes in migration of astrocytes were measured with wound scratch assay and Transwell migration assay. Results Western blot-ting showed that the expression of ski protein was significantly lower in the ski-siRNA group than in the siRNA negative control group and untreated group (F=132.957, P<0.001). Transwell migration assay showed that the number of astrocytes crossing through chambers was less in the ski-siRNA group than in the siRNA negative control group and untreated group (F>47.197, P<0.05). Wound scratch assay showed that the wound healing rate was lower in the ski-siRNA group than in the control group one, two, three, four and five days after transfection (F>69.187, P<0.001). Conclusion Ski knocked down by siRNA could inhibit the migration ability of astrocytes. It is a reminding that ski may take part in the migration process of astrocytes, and moreover, ski may play an important role in the formation of glial scar.
5.Ischemia-free liver transplantation improves the prognosis of recipients using functionally marginal liver grafts
Shuai WANG ; Xiaohong LIN ; Yunhua TANG ; Yichen LIANG ; Min ZHANG ; Zhonghao XIE ; Yiwen GUO ; Yuqi DONG ; Qiang ZHAO ; Zhiyong GUO ; Dongping WANG ; Xiaoshun HE ; Weiqiang JU ; Maogen CHEN
Clinical and Molecular Hepatology 2024;30(3):421-435
Background/Aims:
The shortage of donor liver hinders the development of liver transplantation. This study aimed to clarify the poor outcomes of functionally marginal liver grafts (FMLs) and provide evidence for the improvement of ischemia-free liver transplantation (IFLT) after FML transplantation.
Methods:
Propensity score matching was used to control for confounding factors. The outcomes of the control group and FML group were compared to demonstrate the negative impact of FMLs on liver transplantation patients. We compared the clinical improvements of the different surgical types. To elucidate the underlying mechanism, we conducted bioinformatic analysis based on transcriptome and single-cell profiles.
Results:
FMLs had a significantly greater hazard ratio (HR: 1.969, P=0.018) than did other marginal livers. A worse 90-day survival (Mortality: 12.3% vs. 5.0%, P=0.007) was observed in patients who underwent FML transplantation. Patients who received FMLs had a significant improvement in overall survival after IFLT (Mortality: 10.4% vs 31.3%, P=0.006). Pyroptosis and inflammation were inhibited in patients who underwent IFLT. The infiltration of natural killer cells was lower in liver grafts from these patients. Bulk transcriptome profiles revealed a positive relationship between IL-32 and Caspase 1 (R=0.73, P=0.01) and between IL-32 and Gasdermin D (R=0.84, P=0.0012).
Conclusions
FML is a more important negative prognostic parameter than other marginal liver parameters. IFLT might ameliorate liver injury in FMLs by inhibiting the infiltration of NK cells, consequently leading to the abortion of IL-32, which drives pyroptosis in monocytes and macrophages.
6.LncRNA-m18as1 competitively binds with miR-18a-5p to regulate follicle-stimulating hormone secretion through the Smad2/3 pathway in rat primary pituitary cells.
Weidi ZHANG ; Wenzhi REN ; Dongxu HAN ; Guokun ZHAO ; Haoqi WANG ; Haixiang GUO ; Yi ZHENG ; Zhonghao JI ; Wei GAO ; Bao YUAN
Journal of Zhejiang University. Science. B 2022;23(6):502-514
Long noncoding RNAs (lncRNAs) are expressed in different species and different tissues, and perform different functions, but little is known about their involvement in the synthesis or secretion of follicle-stimulating hormone (FSH). In general, we have revealed lncRNA‒microRNA (miRNA)‒messenger RNA (mRNA) interactions that may play important roles in rat primary pituitary cells. In this study, a new lncRNA was identified for the first time. First, we analyzed the gene expression of lncRNA-m18as1 in different tissues and different stages by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and observed the localization of lncRNA-m18as1 with fluorescence in situ hybridization, which indicated that this lncRNA was distributed mainly in the cytoplasm. Next, we used RT-qPCR and enzyme-linked immunosorbent assay (ELISA) to analyze the regulation of FSH synthesis and secretion after overexpression or knockdown of lncRNA-m18as1 and found that lncRNA-m18as1 was positively correlated with FSH synthesis and secretion. In addition, mothers against decapentaplegic homolog 2 (Smad2) was highly expressed in our sequencing results. We also screened miR-18a-5p from our sequencing results as a miRNA that may bind to lncRNA-m18as1 and Smad2. We used RNA immunoprecipitation-qPCR (RIP-qPCR) and/or dual luciferase assays to confirm that lncRNA-m18as1 interacted with miR-18a-5p and miR-18a-5p interacted with Smad2. Fluorescence in situ hybridization (FISH) showed that lncRNA-m18as1 and miR-18a-5p were localized mainly in the cytoplasm. Finally, we determined the relationship among lncRNA-m18as1, miR-18a-5p, and the Smad2/3 pathway. Overall, we found that lncRNA-m18as1 acts as a molecular sponge of miR-18a-5p to regulate the synthesis and secretion of FSH through the Smad2/3 pathway.
Animals
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Cell Line, Tumor
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Cell Proliferation
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Follicle Stimulating Hormone/metabolism*
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Gene Expression Regulation, Neoplastic
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In Situ Hybridization, Fluorescence
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MicroRNAs/metabolism*
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RNA, Long Noncoding/metabolism*
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Rats