Objective To summarize the role of cellular senescence and senescent secretary phenotype in the intervertebral disc (IVD) degeneration. Methods Relevant articles that discussed the roles of cellular senescence in the IVD degeneration were extensively reviewed, and retrospective and comprehensive analysis was performed. The senescent phenomenon during IVD degeneration, senescent secretary phenotype of the disc cells, senescent pathways within the IVD microenvironment, as well as the anti-senescent approaches for IVD regeneration were systematically reviewed. Results During aging and degeneration, IVD cells gradually and/or prematurely undergo senescence by activating p53-p21-retinoblastoma (RB) or p16INK4A-RB senescent pathways. The accumulation of senescent cells not only decreases the self-renewal ability of IVD, but also deteriorates the disc microenvironment by producing more inflammatory cytokines and matrix degrading enzymes. More specific senescent biomarkers are required to fully understand the phenotype change of senescent disc cells during IVD degeneration. Molecular analysis of the senescent disc cells and their intracellular signaling pathways are needed to get a safer and more efficient anti-senescence strategy for IVD regeneration. Conclusion Cellular senescence is an important mechanism by which IVD cells decrease viability and degenerate biological behaviors, which provide a new thinking to understand the pathogenesis of IVD degeneration.
Objective To investigate the phenotypic change and proliferation of fibroblasts in human inflammatory strictured bile duct wall. Methods We observed the density and ultrastructure of fibroblasts, and the histologic structure in human normal bile duct wall and inflammatory strictured bile duct wall by light and electron microscope.Results The results showed that fibroblasts were the main source of extracellular matrix production in bile duct wall. The phenotype of fibroblasts in inflammatory strictured bile duct wall changed obviously, quiescent fibroblasts were activated and transformed to myofibroblasts, with massive proliferation. Conclusion These data suggest that massive proliferation of activated fibroblasts and myofibroblasts is the main source of extracellular matrix overproduction which results in inflammatory bile duct stricture.
ObjectiveTo explore the difference in clinical characteristics and airway inflammation in COPD patients with different bronchodilator test results. MethodsA total of 237 COPD patients visited between January 2013 and December 2014 were recruited in the study. The ability to complete daily living questionnaire (ADL),modified Medicine Research Council (mMRC) score,6-minute walk distance,pulmonary function,and cell count in induced sputum were measured in the patients. They were divided into a positive group and a negative group according to the response to bronchodilator test and compared. ResultsThere were 58 cases (24.47%) in the positive group,and 179 cases (75.53%) in the negative group. There were no differences in the cumulative amount of smoking[(44.36±17.51) pack-years vs. (50.15±30.51) pack-years],duration of recurrent cough[(14.1±11.1) years vs. (15.5±11.4) years],history of allergic diseases (22.40% vs. 30.80%),or family history of allergic disease (5.17% vs. 2.23%) between two groups. In the positive group,FEV1%pred[(51.04±13.26)% vs. (44.10±14.66)%] and FVC%pred[(73.81±13.60)% vs. (64.33±15.17)%] were better than those of the negative group (both P<0.05). DLCO%pred[(44.66±13.92)% vs. (40.60±17.31)%] and RV/TLC[(51.80±10.57)% vs. (53.16±11.15)%] had no significant differences between two groups. 43.10% of the patients in the positive group and 61.46% in the negative group felt shortness of breath after walking (P<0.05). The positive group scored 22.6±3.8 points in activities of daily living assessment,1.5±0.9 points in mMRC,436.22±102.83 meters in 6-minute walking test,and 2.7±2.1 points in Borg scale score,which were all better than those in the negative group (all P<0.05). There was no significant difference in cell counting in induced sputum between two groups. ConclusionsA part of COPD patients have positive response to bronchodilator,with better lung function,better ADL score,better mMRC score,and farther 6-minute walking distance. It suggests that a positive bronchodilator response might be a clinical phenotype of COPD.
ObjectiveTo investigate the influences of lactic acid (LA), the final degradation product of polylactic acid (PLA) on the prol iferation and osteoblastic phenotype of osteoblast-l ike cells so as to provide theoretical basis for bone tissue engineering. MethodsRos17/2.8 osteoblast-l ike cells were harvested and divided into 3 groups. In groups A and B, the cells were cultured with the medium containing 4, 8, 16, 22, and 27 mmol/L L-LA and D, L-LA, respectively. In group C, the cells were cultured with normal medium (pH7.4). The cell prol iferation was determined with MTT method after 1, 3, and 5 days. The relative growth ratio (RGR) was calculated, and the cytotoxicity was evaluated according to national standard of China. In addition, the alkal ine phosphatase (ALP) activity of cells cultured with medium containing 4 mmol/L L-LA (group A), 4 mmol/ L D, L-LA (group B), and normal medium (group C) after 1 and 5 days were detected with ALP kits, and the relative ALP ratio (RAR) was calculated; after 21 days, the calcium nodules were tested with von Kossa staining method, and were quantitatively analyzed. ResultsWhen LA concentration was 4 mmol/L, the mean RGR of both groups A and B were all above 80%, and the cytotoxic grades were grade 0 or 1, which meant non-cytotoxicity. When LA concentration was 8 mmol/L and 16 mmol/ L, groups A and B showed cytotoxicity after 5 days and 3 days, respectively. When LA concentration was above 22 mmol/L, cell prol iferations of groups A and B were inhibited evidently after 1-day culture. At each LA concentration, RGR of group A was significantly higher than that of group B at the same culture time (P<0.05) except those at 4 mmol/L after 1-day and 3-day culture. After 1 day, the RAR of group A was significantly higher than that of group B on 1 day (144.1%±3.2% vs. 115.2%±9.8%, P<0.05) and on 5 days (129.6%±9.8% vs. 78.2%±6.9%, P<0.05). The results of von Kossa staining showed that the black gobbets in group A were obviously more than those of groups B and C. The staining area of group A (91.2%±8.2%) was significantly higher than that of groups B (50.3%±7.9%) and C (54.2%±8.6%) (P<0.05). ConclusionThe concentration and composition of LA have significant effects on the cell proliferation and osteoblastic phenotype of osteoblast-l ike cells.
ObjectiveTo study the clinicopathological features of mediastinum nodular sclerosis Hodgkin lymphoma (NSHL) in order to improve the recognition of it. MethodsThe clinical data of 3 cases of mediastinum NSHL between 2003 and 2012 were collected. Then we analyzed the carcinoma pathologic samples by pathomorphology, immunophenotypic phenotype, related gene rearrangement and situ hybridization with EBER. ResultsThe pathomorphologic results showed that broad fibrotic bands subdivided the lymphoid parenchyma into large nodules, the tumoral cells had distinct boundary with empty cytoplasm and small-to-medium-sized nucleoli, and the nodules contained inflammatory cell components. The immunophenotypic phenotype of the tumoral cells were CD15, CD30, PAX-5 and CD20 partly, but anaplastic lymphoma kinase, CD45, cytokeratin, CD79α and S-100 were not expressed. T cell receptor γ and IgH gene were no rearranged, and EBER in situ hybridization was not detected. ConclusionVarious lymphomas occur in the mediastinum and mediastinum NSHL is just one of them. Mastering its distinctive pathomorphology and immunophenotypic phenotype is highly significant for diagnosis, differential diagnosis and treatment of the disease.
The aim of this article is to study how andrographolide-releasing collagen scaffolds influence rabbit articular chondrocytes in maintaining their specific phenotype under inflammatory environment. Physical blending combined with vacuum freeze-drying method was utilized to prepare the andrographolide-releasing collagen scaffold. The characteristics of scaffold including its surface morphology and porosity were detected with environmental scanning electron microscope (ESEM) and a density instrument. Then, the release of andrographolide from prepared scaffolds was measured by UV-visible spectroscopy. Rabbit chondrocytes were isolated and cultured in vitro and seeded on andrographolide-releasing collagen scaffolds. Following culture with normal medium for 3 d, seeded chondrocytes were cultured with medium containing interleukin-1 beta (IL-1β) to stimulate inflammation in vitro for 7 d. The proliferation, morphology and gene transcription of tested chondrocytes were detected with Alamar Blue assay, fluorescein diacetate (FDA) staining and reverse-transcriptase quantitative polymerase chain reaction (RT-qPCR) test respectively. The results showed that the collagen scaffolds prepared by vacuum freeze-dry possess a high porosity close to 96%, and well-interconnected chambers around (120.7±17.8) μm. The andrographolide-releasing collagen scaffold continuously released andrographolide to the PBS solution within 15 d, and collagen scaffolds containing 2.22% andrographolide significantly inhibit the proliferation of chondrocytes. Compared with collagen scaffolds, 0.44% andrographolide-containing collagen scaffolds facilitate chondrocytes to keep specific normal morphologies following 7 d IL-1β induction. The results obtained by RT-qPCR confirmed this effect by enhancing the transcription of tissue inhibitor of metalloproteinase-1 (TIMP-1), collagen II (COL II), aggrecan (Aggrecan) and the ratio of COL II/ collagen I(COL I), meanwhile, reversing the promoted transcription of matrix metalloproteinase-1 (MMP-1) and matrix metalloproteinase-13 (MMP-13). In conclusion, our research reveals that andrographolide-releasing (0.44%) collagen scaffolds enhance the ability of chondrocytes to maintain their specific morphologies by up-regulating the transcription of genes like COL II, Aggrecan and TIMP-1, while down-regulating the transcription of genes like MMP-1 and MMP-13 which are bad for phenotypic maintenance under IL-1β simulated inflammatory environment. These results implied the potential use of andrographolide-releasing collagen scaffold in osteoarthritic cartilage repair.
目的 探討不同分子分型乳腺浸潤性導管癌手術病例標本中P53、表皮生長因子受體（EGFR）和Ki-67的表達及臨床意義。 方法 采用免疫組織化學鏈霉菌抗生物素蛋白-過氧化物酶連接法法對2010年1月－2011年7月446例乳腺浸潤性導管癌患者標本進行分子分型，并同時檢測其P53、EGFR、Ki-67等的表達。 結果 P53和Ki-67在人類表皮生長因子受體2（HER2）過表達型、基底細胞樣型、未分類型中的表達明顯強于管腔A型及管腔B型（P＜0.05）；HER2過表達型和未分類型中的EGFR表達明顯強于管腔A型及管腔B型（P＜0.05）。 結論 在使用雌激素受體、c-erbB-2等指標對浸潤性導管癌進行分子分型時同時檢測P53、EGFR及Ki-67等標記物，有助于更加精準的評估腫瘤的生物學行為及預后 ，對靶向藥物的個體化治療提供參考和療效預測有重要意義。
Objective To investigate the potential causal associations between 731 immune cell traits and atherosclerosis by Mendelian randomization (MR) analysis. Methods Using single nucleotide polymorphisms (SNPs) as instrumental variables, genome-wide association study (GWAS) summary statistics (GCST90001391 to GCST90002121) for 731 immune cell traits were obtained from the GWAS Catalog database, and the atherosclerosis dataset (finn-b-I9_CORATHER) was retrieved from the IEU database for MR analysis. The inverse variance weighted method, MR-Egger regression, weighted median, simple mode, and weighted mode approaches were employed to estimate the causal effects between the 731 immune cell traits and atherosclerosis, using odds ratio (OR) with 95% confidence interval (CI) as the effect size. Cochran Q test was used to assess heterogeneity. Horizontal pleiotropy was evaluated using the MR-Egger intercept test and the MR-PRESSO method. Leave-one-out analysis was conducted to examine the sensitivity of the causal estimates to individual SNPs. Results MR analysis revealed potential causal associations between 24 immune cell traits and atherosclerosis (P<0.05). Among them, human leucocyte antigen (HLA)-DR on plasmacytoid dendritic cells (DC) [OR=1.035, 95%CI (1.016, 1.054), P<0.001] and hematopoietic stem cell absolute count (HSCAC) [OR=1.049, 95%CI (1.021, 1.077), P<0.001] showed significant positive causal associations with atherosclerosis (P≤0.001), whereas CD86 on CD62L+ myeloid DC [OR=0.953, 95%CI (0.926, 0.981), P=0.001] exhibited a significant negative causal association with atherosclerosis (P≤0.001). The results of Cochran Q test, MR-Egger regression, and MR-PRESSO indicated P-values>0.05, suggesting no evidence of heterogeneity or horizontal pleiotropy in the causal estimates for these three immune cell traits. Reverse MR analysis, using the 24 immune cell traits as outcome variables, showed no evidence of causal association (P>0.05), supporting a unidirectional causal relationship from immune cells to atherosclerosis. Conclusion HLA-DR on plasmacytoid DC and HSCAC may serve as risk factors for atherosclerosis, while CD86 on CD62L+ myeloid DC may play a protective role against atherosclerosis.
目的 建立急性白血病（AL）患者八色流式免疫表型分析起始管方案。 方法 用胞膜CD3（CD3）、CD19、CD10、CD34、CD45、胞漿CD79a（cCD79a）、髓過氧化物酶（MPO)和胞漿CD3（cCD3）等8種抗體建立八色流式染色方案。膜表面抗體直接染色；膜內抗體經固定破膜，再染色后上機檢測。將3個血小板減少患者骨髓標本分別進行抗體的單色染色和缺一色染色；最后對17例確診的AL初發患者標本進行檢測。 結果 用單色染色來確定染色方案中各抗體的檢測電壓及熒光補償；缺一色染色中，陽性細胞群較單色染色變化均＜10%，表明方案中的各抗體相互作用小。17例AL初發患者中，6例急性B淋巴細胞白血病原始細胞均為CD34和CD19陽性，5例cCD79a陽性和4例CD10陽性；4例急性T淋巴細胞白血病患者均為cCD3陽性；6例急性髓細胞白血病均為CD34和MPO陽性；1例B+T混合表型AL患者CD34、cCD3、CD19、cCD79a及CD10均為陽性，MPO和CD3為陰性，此檢測方案能夠確定各類AL的細胞類型。 結論 建立了AL患者八色流式免疫表型分析起始管方案，操作簡便快速，適用于臨床檢測。