1.An experimental study on the biological characteristics of SDF-1/CS/β-GP composite bio-membrane in vitro
Kaiqi ZHANG ; Jianyong DONG ; Yajie ZHANG ; Yanting HE ; Yuanyuan HUO ; Zhaoyuan LI ; Jun CUI
STOMATOLOGY 2023;43(3):212-217
Objective:
To prepare a composite membrane by chitosan/β-sodium glycerophosphate(CS/β-GP) thermosensitive hydrogel combined with stromal cell derived factor-1(SDF-1) and observe its biological characteristics in vitro.
Methods:
Different doses of SDF-1 were added into CS/β-GP solution and then the thermosensitive gel time was measured. The SDF-1/CS/β-GP solution was membrane paved and dried to prepare composite membranes. The morphological characteristics were observed by scanning electron microscope(SEM). Composite membranes were placed into cell culture medium, and the supernatant(n=3) was extracted after standing at 6, 12, 24, 36, 48, 60 h, respectively. The concentration of SDF-1 in the solution was measured. Bone mesenchymal stem cells(BMSCs) were cultured in the Transwell room, and the composite membranes containing different concentrations of SDF-1 were placed in the lower chamber. There were four groups(n=3): Group M0 used CS/β-GP membrane(control group), Group M1, M2, M3 used SDF-1/CS/β-GP membrane(SDF-1 was 100, 200, 400 ng/mL respectively). After culture for 6, 12 and 24 h, the cells under the membrane were preserved and Giemsa stained and counted. The absorbance(OD) value was measured by MTT method to calculate the cell proliferation rate. SPSS 19.0 was used for multi-factor analysis of variance.
Results :
After adding a certain amount of SDF-1 into CS/β-GP solution, the gel time did not change significantly(P>0.05). The SDF-1/CS/β-GP membrane was translucent and porous at 37 ℃. In this experiment, the volumic mass of SDF-1 released by SDF-1/CS/β-GP composite membrane increased gradually with the experimental time(P<0.01). Transwell cell chemotaxis test showed that the number of BMSCs cells with directional migration increased with the prolongation of observation time(P<0.01) and the increase of SDF-1 volumic mass(P<0.01). In MTT test, the OD value of migration cell solution increased with the prolongation of time(P<0.01) and the increase of SDF-1 volumic mass(P<0.01).
Conclusion
The SDF-1/CS/β-GP composite membrane has a porous structure and biological activity of chemotactic BMSCs directional migration. It is a potential membrane for guided tissue regeneration.
2. Anthrahydroquinone-2,6-disulfonate alleviates paraquat-induced kidney injury via the apelin-APJ pathway in rats
Qi LI ; Tang DENG ; Qi-Feng HUANG ; Shuang-Qin XU ; Hang-Fei WANG ; Xin-Xin WU ; Nan LI ; Yang YI ; Ji-Chao PENG ; Yue HUANG ; Jin QIAN ; Xiao-Ran LIU ; Bo WANG ; Kai-Wen LIN
Asian Pacific Journal of Tropical Biomedicine 2022;12(8):333-342
Objective: To explore the protective effects of anthrahydroquinone-2,6-disulfonate (AH 2 QDS) on the kidneys of paraquat (PQ) poisoned rats via the apelin-APJ pathway. Methods: Male Sprague Dawley rats were divided into four experimental groups: control, PQ, PQ+sivelestat, and PQ+AH 2 QDS. The PQ+sivelestat group served as the positive control group. The model of poisoning was established via intragastric treatment with a 20% PQ pesticide solution at 200 mg/kg. Two hours after poisoning, the PQ+sivelestat group was treated with sivelestat, while the PQ+AH 2 QDS group was given AH 2 QDS. Six rats were selected from each group on the first, third, and seventh days after poisoning and dissected after anesthesia. The PQ content of the kidneys was measured using the sodium disulfite method. Hematoxylin-eosin staining of renal tissues was performed to detect pathological changes. Apelin expression in the renal tissues was detected using immunofluorescence. Western blotting was used to detect the expression levels of the following proteins in the kidney tissues: IL-6, TNF-α, apelin-APJ (the apelin-Angiotensin receptor), NF-κB p65, caspase-1, caspase-8, glucose-regulated protein 78 (GRP78), and the C/EBP homologous protein (CHOP). In in vitro study, a PQ toxicity model was established using human tubular epithelial cells treated with standard PQ. Twenty-four hours after poisoning, sivelestat and AH 2 QDS were administered. The levels of oxidative stress in human renal tubular epithelial cells were assessed using a reactive oxygen species fluorescence probe. Results: The PQ content in the kidney tissues of the PQ group was higher than that of the PQ+AH 2 QDS group. Hematoxylin-eosin staining showed extensive hemorrhage and congestion in the renal parenchyma of the PQ group. Vacuolar degeneration of the renal tubule epithelial cells, deposition of crescent-like red staining material in renal follicles, infiltration by a few inflammatory cells, and a small number of cast formation were also observed. However, these pathological changes were less severe in the PQ+sivelestat group and the PQ+AH 2 QDS group (P<0.05). On the third day after poisoning, immunofluorescence assay showed that the level of apelin in the renal tissues was significantly higher in the PQ+AH 2 QDS group than in the PQ group. Western blotting analysis results showed that IL-6, TNF-α, NF-κB p65, caspase-1, caspase-8, GRP78, and CHOP protein levels in the PQ group were higher than in the PQ+AH 2 QDS group (P<0.05). The expression of apelin-APJ proteins in the PQ+AH 2 QDS group was higher than in the PQ+sivelestat and PQ groups (P<0.05); this difference was significant on Day 3 and Day 7. The level of oxidative stress in the renal tubular epithelial cells of the PQ+AH 2 QDS group and the PQ+sivelestat group was significantly lower than in the PQ group (P<0.05). Conclusions: This study confirms that AH 2 QDS has a protective effect on PQ-poisoned kidneys and its positive effect is superior to that of sivelestat. The mechanism of the protective effects of AH 2 QDS may be linked to reduction in cellular oxidative stress, PQ content of renal tissue, inflammatory injury, endoplasmic reticulum stress, and apoptosis. AH 2 QDS may play a role in the treatment of PQ poisoning by upregulating the expression of the apelin-APJ.
3.International and domestic researches about neurogenic bladder: a visualized analysis
Xiangzhi MENG ; Shenhong CUI ; Xiaoqian HOU ; Benyuan LI ; Xinru ZHANG ; Ping ZHANG ; Yunbo HAN ; Jun LENG
Chinese Journal of Rehabilitation Theory and Practice 2022;28(4):439-446
Objective To evaluate the development, hot spots and trends of the fields of neurogenic bladder.Methods The relevant articles of neurogenic bladder from January, 2000 to June, 2021 in CNKI and Web of Science were retrieved.The countries, authors, institutions, cited reference and keywords were extracted with CiteSpace to draw knowledge mapping. Results and Conclusion A total of 5 064 articles were enrolled. At present, the research on the field of neurogenic bladder is in a stable period of development, and this field has been widely concerned by scholars at home and abroad. The cooperation between domestic authors and institutions is not close enough compared with foreign countries, and domestic cooperation is more between medical schools and their respective affiliated hospitals. In the future, China can further strengthen cross-regional and cross-agency cooperation. Low-frequency electrical stimulation and sacral nerve regulation are seem to be research hotspots, and children39;s neurogenic bladder and robot-assisted technologies are also needed more attention.
4.Expression change of miR-96 in endometrial cancer and its effect on malignant biological behaviors of cancer cells
LI Feng1a, ; LIU Zhuo2 ; ZHANG Hongping1b
Chinese Journal of Cancer Biotherapy 2021;28(3):275-282
[Abstract] Objective: To investigate the expression changes of miR-96 in endometrial cancer tissues and cells, and to explore its effect on tumor malignant phenotypes as well as the possible mechanisms. Methods:From April 2016 to December 2018, 76 cases of endometrial cancer tissues from 76 patients who were surgically treated in the Department of Obstetrics and Gynecology of our hospital were selected for this study. qPCR was used to detect the expression of miR-96 in human endometrial cancer tissues and cells, and the correlation between the miR-96 expression and the clinicopathological characteristics of patients was analyzed. miR-96 inhibitor was transfected into human endometrial cancer Ishikawa cells in vitro. After transfection, the expression of miR-96 in Ishikawa cells was detected by qPCR; the tumor biological behaviors of Ishikawa cells were detected by CCK-8 test, Clone formation test, Flow cytometry, Scratch test and Transwell test; and the FOXO1 protein expression in Ishikawa cells was detected by WB. At the same time, Dual luciferase reporter gene assay was used to observe the targeting relationship between miR-96 and FOXO1. Results: The results of qPCR showed that the expression of miR-96 was abnormally high in human endometrial cancer cells (JEC, Ishikawa, HEC-1B) and endometrial cancer tissues (all P<0.01), and the expression of miR-96 was closely related to FIGO stage and lymph node metastasis (all P<0.05). After transfection with miR-96 inhibitor, the expression level of miR-96 in Ishikawa cells decreased significantly (P<0.01), the proliferation activity and clone formation ability decreased significantly (all P<0.01), the apoptotic rate increased significantly (P<0.01), and the scratch healing rate and the number of invasive transmembrane cells decreased significantly (P<0.01). Dual luciferase reporter gene assay showed that miR-96 could directly target FOXO1, and WB showed that miR-96 could negatively regulate FOXO1 protein expression in Ishikawa cells (P<0.01). Conclusion: The expression of miR-96 is abnormally high in endometrial cancer tissues and cells. Inhibiting the expression of miR-96 can inhibit the proliferation, invasion and migration of endometrial cancer cells and promote their apoptosis. The mechanism may be related to the targeted regulation of FOXO1.

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