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Objective: To investigate the effects of Akkermansia muciniphila ( A. muciniphila ) on the proliferation, apoptosis and insulin secretion of rat pancreatic islet cell tumor cells (INS-1)., Methods: INS-1 cells were divided into three groups, normal, repair, and protect groups, and subsequently every group was subjected with A. muciniphila metabolites, live A. muciniphila orpasteurized A. muciniphila for 48 h. A group that did not treat with anything was set as blank control. After intervention, the cell viability was determined by MTT method, the insulin secretion level stimulated by glucose was determined by ELISA, the expressions of the genes involved in insulin secretion and apoptosis were tested by qRT-PCR, and the expression of apoptosis related protein Bax was evaluated by Western blot., Results: There was no significant change in INS-1 cell morphology after co-incubation with 3 types of A. Muciniphila interventions for 48 h. The proliferative activity of INS-1 cells was decreased in the repair group that treated with live A. muciniphila than that of control ( P <0.005). A. muciniphila intervention had no effect on insulin secretion in INS-1 cells in normal, repair or protection group ( P >0.05). A. muciniphila secretions promoted the expression of glucose transporter 2 ( Glut 2) in 3 groups and the expression of glucokinase ( GCK ) in repair group ( P <0.05). The expression of Bax of the INS-1 cell in the normal group was decreased after intervented with 3 kinds of A. muciniphila intervention materials ( P <0.001).The expression of Bax gene of the INS-1 cell in the repair group that treated with dead A. muciniphila was decreased ( P <0.05). The expression of Bax protein of INS-1 cells that treated with A. muciniphila interventions was decreased., Conclusion: A. muciniphila can promote the expression of insulin secretion-related genes in INS-1 cells, inhibit the expression of apoptotic genes and apoptosis protein Bax.This research provides a new direction for applying A. muciniphila in improving type 2 diabetes., (Copyright© by Editorial Board of Journal of Sichuan University (Medical Science Edition).)

Objective: To investigate the effect of estradiol (E2)/estrogen receptor 1 (ESR1) on the proliferation of human chondrocytes in vitro and explore the molecular mechanism., Methods: The Ad-Easy adenovirus packaging system was used to construct and package the ESR1-overexpressing adenovirus Ad-ESR1. Western blotting and qPCR were used to detect the expression of ESR1 protein and mRNA in human chondrocyte C28I2 cells. In the cells treated with different adenoviruses, the effects of E2 were tested on the expressions of proteins related with cell autophagy and apoptosis and the phosphorylation of ERK signaling pathway using Western blotting. Immunofluorescence assay was used to observe the intracellular autophagic flow, flow cytometry was performed to analyze the cell apoptosis rate and the cell cycle changes, and qPCR was used to detect the expressions of PCNA, cyclin B1 and cyclin D1 mRNAs. The inhibitory effect of the specific inhibitor of ERK on the expressions of autophagy- and apoptosis-related genes at both the protein and mRNA levels were detected using Western blotting and qPCR., Results: Transfection with the recombinant adenovirus overexpressing ESR1 and E2 treatment of C28I2 cells significantly enhanced the expressions of autophagy-related proteins LC3, ATG7, promoted the colocalization of LC3 and LAMP1 in the cytoplasm, increased the expressions of the proliferation-related marker genes PCNA, cyclin B1 and cyclin D1, and supressed the expressions of cleaved caspase-3, caspase-12 and pERK. RNA interference of ESR1 obviously lowered the expression levels of autophagy-related proteins in C28I2 cells, causing also suppression of the autophagic flow, increments of the expressions of apoptosis-related proteins and pERK, and down-regulated the expressions of the proliferation marker genes. Blocking ERK activation with the ERK inhibitor obviously inhibited the effects of E2/ESR1 on autophagy, proliferationrelated gene expressions and cell apoptosis., Conclusions: The targeted binding of E2 with ESR1 promotes the proliferation of human chondrocytes in vitro possibly by inhibiting the activation of ERK signaling pathway to promote cell autophagy and induce cell apoptosis.

Purpose: To investigate the effects of curcumin and paclitaxel on proliferation and apoptosis of human oral squamous carcinoma cell line CAL27., Methods: CAL27 cells were cultured and treated with different concentrations of curcumin and paclitaxel separately or in combination for 24, 48 and 72 h. The cell proliferation was evaluated by MTT assay, and cell apoptosis was detected by Tunel staining. Expression of Bcl-2, Bax, and active caspase-3 were analyzed by Western blot. The data were analyzed with SPSS 11.0 software package., Results: Compared with the effects of curcumin or paclitaxel, combined treatment of both drugs significantly inhibited cell growth, induced cell apoptosis, decreased the expression of Bcl-2 and the ratio of Bcl-2/Bax, and increased the expression of Bax and active caspase-3 in CAL 27 cells., Conclusions: Combined treatment of Curcumin and Paclitaxel significantly inhibits cell growth and mediated cell apoptosis compared with that of either single drug.

This study was aimed to explore the change of aggregation and activation of platelets loaded with epigallocatechin gallate (EGCG). The platelets were treated by loading buffer with different concentrations of EGCG (0, 2.5, 5, 7.5, 10, 12.5, 15, 20 and 30 mmol/L) and were divided into 2.5, 5, 10 mmol/L groups and control group. The physiological and biochemical functions of platelets were observed, including recovery rate, aggregation and activation of platelets. The platelet counts were determined by Counter Cell-DYN 1200. The aggregation activities were tested through turbidimetry, the platelet apoptosis was detected by flow cytometry. The results showed that the concentrations of EGCG loading in platelets of 2.5, 5 and 10 mmol/L groups were 0.4006 ± 0.12, 1.0527 ± 0.1503, 1.6902 ± 0.1112 mmol/L respectively. Along with the increasing of EGCG concentrations in loading-buffer, the EGCG absorbed by platelets increased too. When the concentration of EGCG in loading-buffer exceeded 15 mmol/L, the EGCG absorbed by platelets did not increase. The recovery rate in 2.5 mmol/L loading buffer group was 82.45 ± 0.360% which was lower than that in control group (90.33 ± 1.115%) (p < 0.05). As compared with control group, the recovery rate in 5 mmol/L loading buffer group (57.51 ± 2.468)% and 10 mmol/L loading buffer group (47.45 ± 2.030)% were even significantly lower (p < 0.01). When ADP was used as the inducer, the maximal aggregation rate (MAR) in control group was (63.6 ± 4.037)%, which was higher than that in other EGCG-loading groups (p < 0.01). And the aggregation activity of platelets negatively correlated with the concentration of EGCG in loading-buffer. When THR was used as the inducer, the MAR in control group was (89.3 ± 6.533)% and higher than that those in other groups (p < 0.05), especially in groups with loading-buffer higher than 10 mmol/L EGCG (70.1 ± 5.400%) (p < 0.01). In the experiment of cellular apoptosis, the early apoptosis easy appeared in platelets loaded with EGCG. It is concluded that the EGCG loading in platelets markedly influences the physiological and biochemical functions of platelets, the apoptosis easy occurs in platelets loaded with EGCG. The EGCG accelerates the course of platelet apoptosis.

Objective: To investigate and analyze the changes of neuronal-apoptosis and the expression of caspase-3 for finding out a new method of injury timing after brain contusion in human., Methods: The tissue was stained by TUNEL for apoptosis and by immunohistochemistry for caspase-3. Image analysis technique was employed., Results: The nerve cells stained positive by TUNEL and Caspase-3 immunohistochemistry were distributed in the penumbra and central area. Both these areas were in striking contrast with the distal area or those of control group. The positive staining was more prominente in penumbra area than in central area (P < 0.05). The changes of TUNEL staining and expression of Caspase-3 in penumbra area gradually increased with the survival period after injury; they were parallel to each other. There were linear relationships between the time of injury in 48 hours and the increase in the mean of integral optical density (IOD), the coefficient of correlation (r) being 0.93 and 0.69 for the two staining methods, and two linear regression formulae were induced, respectively., Conclusion: Observations on the increasing of neuronal apoptosis and Caspase-3 expression in relation with the survival period after injury could be utilized in the timing of brain contusion.

Objective: To investigate the effect of two core binding factors alpha 1 (Cbfa1) isfroms (Cbfa1/P56 and Cbfa1/P57) on the apoptosis of mesenchymal cell line MBA-1., Methods: The two Cbfal isfroms were transiently transfected into MBA-1 cells, then the changes of apoptosis rate were observed by flow cytometer. The protein expressions of Cbfa1, Bax, Bcl-2, caspase-3, and caspase-9, cytochrome-C and TNF-alpha were determined by Western immunoblot., Results: After the transient transfection with the two isforms of Cbfa1, MBA-1, the cells apoptotic rates increased, and the ratio of Bax/Bcl-2, the expressions of cytochrome-C, caspase-3, caspase-9, and TNF-alpha were significantly increased., Conclusion: Cbfa1 can promote the apoptosis in mesenchymal cell line MBA-1. Bax/Bcl-2, cytochrome-C, caspase-9, caspase-3, and TNF-alpha are also involved in the apoptosis pathway.

Objective: To inquire into the pathologic diagnosis and the dating of injuries of light closed encephalon injury., Methods: Wistar rats were hurt by fluid percussion, and were killed respectively at 15 min, 30 min, 1 h, 3 h, 6 h, 12 h, 1 d, 4 d, 7 d, and 14 d after injury. The expression of Fas in the cerebral cortex, thalamus, and hippocampi was detected by immunohistochemistry and the results were assessed by image analysis system., Results: The results showed that the expression of Fas could be detected in thirty minutes after injury, and then it increased significantly in three hours, reached apex in twelve hours after injury, decreased gradually in four days after injury, and returned to normal in 14 days after injury., Conclusion: This research demonstrated that Fas-mediated apoptosis appeared not only around brain trauma but also in the brain tissue far away from the traumatic area. It indicted that Fas expression is a useful target for diagnosis of early brain injury and the regularity of Fas expression could be used as one of indications in deducing the time of brain injury.

Puerins, a kind of nitrogen-containing polyphenol compound with unique structure, is a kind of catechin derivative first isolated from ripened Pu-erh tea. In this paper, the effects of puerins V-VII on breast cancer cells were studied. Four breast cancer cell lines with different receptor phenotypes were used as models, and MTT assay, proteomics detection and apoptosis detection were carried out. The results show that all puerins V, VI and VII had obvious inhibitory effects on the four kinds of breast cancer cells. Proteomics analysis indicates that the differentially expressed proteins were mainly concentrated in the metabolic pathway, followed by cancer pathway and adhesion spot signal pathway under the intervention of puerins V-VII. Apoptosis detection demonstrates that all three puerins compounds could induce the apoptosis of MDA-MB-231 cells. This study first reported the anti-breast cancer activity and preliminary mechanism of puerins V-VII, which provided basic information for studying the anti-cancer active substances of ripened Pu-erh tea. [ABSTRACT FROM AUTHOR]

Objective To investigate the expression of lncRNA SENCR in aortic dissection (AD) tissues of AD patients and its effect on and mechanism in the proliferation apoptosis of human vascular smooth muscle cells (HVSMCs). Methods HE staining was done to detect the pathological changes of AD tissues. Fluorescence in situ hybridization (FISH) and RT-qPCR were used to determine the expression of SENCR in the AD tissue and HVSMCs and the expression of SENCR and miR-206 in the tissues, respectively. HVSMCs were cultured and trans-fected with pcDNA3.1-SENCR overexpression plasmids, or pcDNA3.1 blank plasmid. Then cell proliferation and apoptosis were detected by CCK-8 method and Annexin V/PI double staining flow cytometry assay, respectively. Double luciferase report verified the targeting relationship between SENCR and miR-206. Results SENCR was mainly located in the cytoplasm and nucleus of HVSMCs. Compared with the normal tissue, the expression of SENCR in the AD tissues was down-regulated (P < 0.01), but the expression of miR-206 was up-regulated (P < 0.01). Overexpressed SENCR decreased the cell proliferation of HVSMCs (P < 0.01), but significantly increased the cell apoptosis of HVSMCs (P < 0.01). SENCR could target and negatively regulate miR-206. Conclusion The expression of SENCR is down-regulated in AD tissues, and overexpressed SENCR may inhibit the proliferation and promote the apoptosis of HVSMCs by targeting down-regulated miR-206. [ABSTRACT FROM AUTHOR]

Objective To analyze the expression of immune cells and immunosuppressive factors in the immune microenvironment of glioblastoma. Methods A total of 30 glioma specimens, all of which were glioblastoma (IDH - wildtype), were surgically removed and completely preserved in the First Affiliated Hospital of Ji'nan University and Guangdong Sanjiu Brain Hospital from November 2020 to April 2021. T lymphocytes (CD3+ T cells, CD4+ T cells, CD8+ T cells), suppressive immune cells [FoxP3+ regulatory T cell (Treg)], immunosuppressive factors [transforming growth factor - β (TGF - β)], immunosuppressive factors [programmed cell death protein ligand 1 (PDL1)] in the immune microenvironment of glioblastoma were detected by immunohistochemistry. Results The proportion of CD4+ T cells in glioblastoma was very low. CD3+ T cells accounted for more than 3% in 15 cases (50%), and CD8+ T cells were similar to CD3+ T cells. Few FoxP3+ Treg cells were detected in only 2 cases (6.67%). The cytoplasm of 24 cases (80%) of glioblastoma showed strong positive expression of TGF - β, and its expression was correlated with the distribution of CD3+ T cells. No CD3+ T cells were found in the areas with high expression of TGF-β, and CD3+ T cells were widely distributed in the areas with low expression of TGF-β. PDL1 expression was non- detected. Conclusions TGF - β protein was highly expressed in glioblastoma, and future drugs or immunotherapies designed to target TGF-β may contribute to the clinical treatment of glioblastoma. [ABSTRACT FROM AUTHOR]

Objective To investigate the effects of growth differentiation factor-11 (GDF-11) on bone marrow endothelial progenitor cells (BM-EPC) cultured with high glucose and its possible mechanism. Methods BM-EPC were cultured in vitro and divided into the following groups: normal glucose (NG) group, high glucose (HG) group, HG+recombinant GDF-11 (rGDF-11) group and HG+rGDF-11+TGF-β receptor I inhibitor SB431542 group. Proliferation was assessed by MTT, and apoptosis of EPC were stained with Annexin V-FITC and propidium iodide (PI), and assessed by flow cytometry. The expression of Bcl-2, Bax and Cleaved-caspase 3 were detected by western blot. The phosphorylation of Smad were assessed by immunofluorescence. Data among multiple groups were compared by using one-way ANOVA and the least significant difference t test. Results Compared with the vehicle control group, Bcl-2/Bax ratio decreased, the protein levels of Cleaved-caspase 3 increased and the percentage of apoptotic EPC elevated in those groups cultured with HG [0.45±0.09 vs 1.21±0.23, 0.28±0.02 vs 0.07±0.00, (18.63±3.02)% vs (2.82±0.46)%, t=3.04, -68.06, -7.60, all P<0.05]. However, the effects were blocked by pretreated with rGDF-11. Treatment of EPC with TGF-β receptor I inhibitor SB431542 partially abolished the pro-proliferation and anti-apoptotic effects of GDF-11 (0.56 ± 0.11 vs 1.14 ± 0.07, t=4.33, P<0.05). Immunofluorescence analyses showed that Smad2/3 phosphorylation decreased in high glucose culturing. Pre-incubation with rGDF-11 restored p-Smad2 / 3 expression, however, GDF-11 mediated Smad2 / 3 activation was partially blocked by SB431542 (0.89±0.12 vs 1.21±0.12, t=-3.20,P<0.05). Conclusion GDF-11 may prevent high glucose-induced EPC apoptosis and improve proliferation through activation of TGF-β/Smad2/3 signaling pathway. [ABSTRACT FROM AUTHOR]