Objective To observe the value of serum soluble receptor of advanced glycation endproducts (sRAGE) combined with lung function and high resolution lung CT (HRCT) in predicting the risk of chronic obstructive pulmonary disease (COPD) developing non-small cell lung cancer (NSCLC). Methods From January 2019 to June 2021, 140 patients with COPD combined with NSCLC, 137 patients with COPD, and 133 patients with NSCLC were enrolled in the study from the People's Hospital of Ningxia Hui Autonomous Region. General data, clinical symptoms, pulmonary function indexes and HRCT emphysema indexes (EI) were collected. Serum sRAGE levels of these patients were measured by enzyme linked immunosorbent assay. Clinical characteristics of patients with COPD complicated with NSCLC were analyzed. Serum sRAGE, lung function and lung HRCT were combined to evaluate the correlation between the degree of emphysema and the occurrence of NSCLC in COPD, and receiver operator characteristic (ROC) curve analysis was performed for diagnostic efficiency. Results Compared with NSCLC group, COPD combined with NSCLC group had higher proportion of male patients, higher proportion of elderly patients, higher smoking index, and higher proportion of squamous cell carcinoma (P<0.05). FEV1 and FEV1%pred in COPD combined with NSCLC group were significantly lower than those in COPD group and NSCLC group. The Goddard score and EI values of emphysema were significantly increased (P<0.05). Serum sRAGE was significantly lower than that of COPD group and NSCLC group (P<0.05). Serum sRAGE level was positively correlated with FEV1%pred (r=0.366, P<0.001) and FEV1/FVC (r=0.419, P<0.001), and negatively correlated with Goddard score (r=–0.710, P=0.001) and EI value (r=–0.515, P<0.001). Binary multi-factor logistic regression analysis showed that age, smoking index, EI, Goddard score, RV/TLC were positively correlated with the risk of COPD developing NSCLC, while FEV1%pred, FVC, FEV1/FVC and serum sRAGE were negatively correlated with the risk of COPD developing NSCLC. ROC curve results showed that the area under the curve (AUC) of single diagnosis of sRAGE was 0.990, and the optimal cut-off value of 391.98 pg/mL with sensitivity of 93.3% and specificity of 89.7%. The AUC of sRAGE combined with age, smoking index, EI, Goddard score, FEV1%pred, FVC, FEV1/FVC, RV/TLC was 1.000 with sensitivity of 96.7%, specificity of 96.6%, and Yoden index of 0.933. Conclusion The combination of serum sRAGE, lung function and HRCT emphysema score can improve prediction of NSCLC occurrence in COPD.
Objective To investigate the effect of CD44 fucosylation on fluid adhesion force of rabbit bone marrow mesenchymal stem cells (BMSCs). Methods The rabbit BMSCs were isolated and purified by density gradient centrifugation combined with adherent culture method. The morphology of cells were observed by inverted microscope, and the cell surface markers of CD44, CD34, CD29, and CD105 were assessed by flow cytometry. BMSCs fucosylated by alpha-(1, 3)-fucosyltransferase Ⅵ (FTⅥ) were as the experimental group, and the non-fucosylated BMSCs were as the control group, and then the positive rate of sialyl-LewisX (sLeX) and the binding rate of E-selectin were detected by flow cytometry. The fucosylated BMSCs resuspended in Hank balanced salt solution (HBSS) were assigned as the experimental group (group A), at same time, the non-fucosylated BMSCs resuspended in HBSS solution as the study control group (group B), and the fucosylated BMSCs resuspended in HBSS solution which was added EDTA as negative control group (group C). The fluid adhesion force of rabbit BMSCs were detected by the parallel flow chamber adhesion test. Results Primary BMSCs mainly shaped as spindle and kept strong growth. The third generation BMSCs were negative for CD34, but positive for CD44, CD29, and CD105. After fucosylation, the positive rate of sLeX in the experimental group was 32.52%±1.76%, which was significantly higher than that in the control group (1.48%±0.51%) (t=29.277, P= 0.000). The binding rate of E-selectin in the experimental group was 41.05%±1.84%, which was also significantly higher than that in the control group (4.33%±0.92%) (t=35.674, P=0.000). With the increase of fluid shear force, the number of BMSCs adhering to the surface of human umbilical vascular endothelial cells (HUVEC) in group A was increased at first and then decreased, while there was few BMSCs adhering to the surface of HUVEC in groups B and C. Under the different fluid shear stress, the number of BMSCs adhered to the surface of HUVEC in group A was significantly higher than that in groups B and C (P<0.05), and there was no significant difference between groups B and C (P>0.05). Conclusion CD44 fucosylation on BMSCs can enhance the fluid adhesion force of rabbit BMSCs.
O-linked N-acetylglucosamine (O-GlcNAc) glycosylation is an important form of post-translational protein modification, mainly intracellular. It is closely related to cellular signaling pathways, and is involved in signal transduction, gene transcription and other important biological processes. Studies have found that O-GlcNAc glycosylation is directly related with diabetic retinopathy (DR), further studies may help us to uncover the DR mechanism, and develop new strategies for the diagnosis and treatment of this disease.
ObjectiveTo observe the effect of polypyramidine tract binding protein-associated splicing factor (PSF) towards advanced glycation end products (AGEs) induced the apoptosis of Müller cells in vitro.MethodsExperimental study. Müller cells were cultured and divided into groups according to the project design, plasmid enhanced green fluorescent protein-PSF were transfected into the cells to achieve the overexpression of PSF Müller cells in vitro, then cells were exposed to AGEs and the Morphological changes were observed by HE staining and Hoechst 33258 staining while the survival rate of cells were detected by MTT assay. The effects of PSF on AGEs-induced Müller apoptosis was measured by Cell Death Detection ELISA kit. Meanwhile, 2′,7′-dichlorofluorescin diacetate staining was performed to monitor the protective effects of PSF on AGEs-induced Müller cells ROS.ResultsThe morphology of cells in normal group was full and the cytoplasm staining was uniform. In N+AGEs group and Vec+AGEs group, cell volume decreased, cytoplasm was dense and concentrated, and eosinophilic staining was enhanced. The cell morphology of PSF+AGEs group was still full, with uniform cytoplasm staining and uniform nucleus staining. The viability of N+AGEs group, Vec+AGEs group and PSF+AGEs group were 0.42±0.11, 0.35±0.12 and 0.68±0.12. The apoptosis values were 1.08±0.16, 0.96±0.20 and 0.44±0.08. The intracellular ROS levels were 28 833.67±3 550.06, 28 356.67±4 854.81, 186 163.00±382.54. Compared with N+AGEs group and Vec+AGEs group, the cell viability of PSF+AGEs group was significantly improved (F=20.65, P=0.000), cell apoptosis value (F=43.43, P=0.000) and intracellular ROS level (F=18.86, P=0.000).ConclusionPSF overexpression play a protective role in AGEs-induced apoptosis by inhibiting the production of ROS in Müller cells.
Obiective lt;brgt;To investigate the change of the activity of proliferation in cultivated Muuml;ller cells treated by advanced glycation endoproducts (AGEs), and the effect of these changes on expression of occludin in bovine retinal vascular endothelial cells (BREC). lt;brgt;Methods lt;brgt;The cultivated Muuml;ller cells were devided into normal growth group and cultured with AGEs group. The cultured BREC were devided into 4 groups:group 1, without any medium; group 2, with normal growth Muuml;ller cell medium (MCM); group 3,MCM treated by AGEs; group 4, without cell as the control. Enzyme-linked immuno sorbent assay was used to detect the content of occludin in the medium in the 4 groups. lt;brgt;Results lt;brgt;The content of expression of occludin was the most in group 2, less in group 1, and the least in group 3. lt;brgt;Conclusion lt;brgt;AGEs may promote the abnormal proliferation of Muuml;ller cells and inhibit the expression of occludin in BREC. lt;brgt;(Chin J Ocul Fundus Dis, 2006, 22: 28-30)
Objective To investigate the effects of advanced glycation endproducts (AGEs) on proliferation of pericytes of bovine retinal capillary vessels and expression of transforming growth factor beta;(TGF-beta;). Methods The proliferation of pericytes detected by methyl thiazolyl tetrazolium (MTT) colorimetric assay, cellular cycle of pericytes was analyzed by flow cytometry was used to analyze cell, and TGF-beta; protein expression of pericytes was observed by immunofluorescent staining. Results AGEs inhibited the proliferation of pericytes of bovine retinal capillary vessels, stopped the cellular cycle of pericytes in synthesis phase (S phase), increased the number of apoptotic cells obviously (Plt;0.01), and promoted the expression of TGF-beta; proteinof perycytes. Conclusions AGEs may promote the apoptosis of pericytes by inhibiting the proliferation of pericytes to lead the decrease of pericytes number, and may accelerate diabetic retinopathy by promoting the expression of TGF-beta; protein of pericytes. (Chin J Ocul Fundus Dis, 2006, 22: 20-23)
ObjectiveTo summarize the research progress of the effects of high glucose microenvironment on the biological activity of adipose-derived stem cells (ADSCs).MethodsThe literature on the high glucose microenvironment and ADSCs at home and abroad in recent years was reviewed, and the effects of high glucose microenvironment on the general characteristics, differentiation potential, angiogenesis, and nerve regeneration of ADSCs were summarized.ResultsThe accumulation of advanced glycosylation end products (AGEs) in the high glucose microenvironment led to changes in the biological activities of ADSCs through various pathways, including cell surface markers, proliferation, migration, multi-lineage differentiation, secretory function, and tissue repair ability. The ability of ADSCs to promote angiogenesis and nerve regeneration in high glucose microenvironment is still controversial.ConclusionHigh glucose microenvironment can affect the biological activity of ADSCs, and the effect and mechanism of ADSCs on angiogenesis and nerve regeneration in high glucose microenvironment need to be further studied.
Cell migration is defined as the directional movement of cells toward a specific chemical concentration gradient, which plays a crucial role in embryo development, wound healing and tumor metastasis. However, current research methods showed low flux and are only suitable for single-factor assessment, and it was difficult to comprehensively consider the effects of other parameters such as different concentration gradients on cell migration behavior. In this paper, a four-channel microfluidic chip was designed. Its characteristics were as follows: it relied on laminar flow and diffusion mechanisms to establish and maintain a concentration gradient; it was suitable for observation of cell migration in different concentration gradient environment under a single microscope field; four cell isolation zones (20 μm width) were integrated into the microfluidic device to calibrate the initial cell position, which ensured the accuracy of the experimental results. In particular, we used COMSOL Multiphysics software to simulate the structure of the chip, which demonstrated the necessity of designing S-shaped microchannel and horizontal pressure balance channel to maintain concentration gradient. Finally, neutrophils were incubated with advanced glycation end products (AGEs, 0, 0.2, 0.5, 1.0 μmol·L−1), which were closely related to diabetes mellitus and its complications. The migration behavior of incubated neutrophils was studied in the 100 nmol·L−1 of chemokine (N-formylmethionyl-leucyl-phenyl-alanine) concentration gradient. The results prove the reliability and practicability of the microfluidic chip.
ObjectiveTo observe and analyze the effect of HbA1c level on macular microcirculation in patients with type 2 diabetes mellitus (T2DM).MethodsA cross-sectional study. One hundred and twenty-four T2DM patients (124 eyes) without diabetic retinopathy who diagnosed by the examination of fundus color photography in Lixiang Eye Hospital Of Soochow University during September to December 2017 were enrolled in this study. There were 59 males (59 eyes) and 65 females (65 eyes), with the mean age of 65.06±7.99 years old. All patients underwent BCVA, fundus color photography, and OCT angiography (OCTA). The history of diabetes, hypertension and dyslipidemia were recorded in detail. According to the HbA1c level, patients were divided into three groups, HbA1c ideal control group (group A, HbA1c ＜7%, 67 eyes), HbA1c control group (group B, 7%≤HbA1c≤9%, 44 eyes), and HbA1c poor control group (group C, HbA1c＞9%, 13 eyes), respectively. The 3 mm×3 mm range of the macular area was scanned by OCTA instrument. The vascular density (VD) and skeleton density (SD) of nonsegmented retinal layer (NRL), superficial retinal layer (SRL) and deep retinal layer (DRL) in the macular area and foveal avascular zone (FAZ) area, non-circularity index, axial rate (AR) of SRL were measured. The correlation between HbA1c, BCVA and VD, SD of NRL, SRL, DRL was analyzed statistically with Spearman correlation test. The correlation between systemic factors and the above indicators was analyzed statistically with linear regression analysis.ResultsThe results of linear regression analysis showed that HbA1c was significantly correlated with VD (t=−3.237, −3.156, −2.050) and SD (t=−0.3.45, −3.034, −2.248) of NRL, SRL and DRL (P＜0.05); but no correlation with FAZ, non-circularity index and AR (t=1.739, 0.429, 1.155; P＞0.05). The differences of VD (F=6.349, 5.981, 3.709), SD (F=7.275, 6.085, 1.904) and AR (F=0.027) of NRL, SRL and DRL in group A, B and C were statistically significant (P＜0.05); but the differences of FAZ (F=1.904), non-circularity index (F=0.280) was not statistically significant (P＞0.05). Significant differences (P＜0.05) of VD and SD of NRL were found between group A and B (t=1.987, 2.201), group A and C (t=3.365, 3.572), group B and C (t=2.010, 2.076). Significant differences (P＜0.05) of VD and SD of SRL were found between group A and B (t=2.087, 2.168), group A and C (t=3.197, 3.194). There were significant differences (P＜0.05) in SD of DRL between group A and B (t=2.239), group A and C (t=−2.519). There was significant difference in VD of DRL between group A and C (t=2.363). The results of Spearman correlation analysis showed that HbA1c was negatively correlated with VD (r=−0.273, −0.255, −0.222; P=0.002, 0.004, 0.013) and SD (r=−0.275, −0.236, −0.254; P＜0.05) of NRL, SRL, DRL; positively correlated with FAZ and BCVA (r=0.221, 0.183; P＜0.05). BCVA was negatively correlated with VD (r=−0.210, −0.190, −0.245) and SD (r=−0.239, −0.207, −0.296) of NRL, SRL, and DRL (P＜0.05), but not correlated with FAZ (r=0.099, P＞0.05).ConclusionThe decrease of macular perfusion and the morphological change of FAZ accompanied by HbA1c increased.