Bone marrow mesenchymal stem cells interactions with hepatocytes and hepatic stellate cells on different stiff substrates / 中华肝脏病杂志

Objective@#To explore the effect of substrate mechanical microenvironment and cell-cell interaction on differentiation of bone marrow mesenchymal stem cells (BMSCs), intrahepatic cellular function and phenotype.@*Methods@#Bone marrow mesenchymal stem cells (BMSCs)-hepatocytes (HCs) and BMSCs-hepatic stellate cells (HSCs) were co-cultured on polyvinyl alcohol (PVA) hydrogel substrates at different stiffness (4.50 ± 0.47 kPa, 19.00 ± 3.51 kPa and 37.00 ± 2.09 kPa) by non-contact co-culture method. Furthermore, the effect of substrate mechanical microenvironment on BMSCs, HCs and HSCs and the activation and proliferation of HCs under different co-cultured condition was studied. A Student's t-test was used to compare the two groups.@*Results@#The expression ofα-smooth muscle actin (α-SMA) and collagenα1- I (Col1A1) in BMSCs and HSCs cultured on its own increased with increase of substrate stiffness. After 72 h, the expression of albumin (ALB) of HCs on three stiff substrates was significantly higher than that of 24 and 48 h. Moreover, the expression of ALB of HCs increased with the increase of substrate stiffness. During the co-culture of BMSCs and HSCs, BMSCs of all three stiffness substrates promoted the expression ofα-SMA, Col1A1 in HSCs, but reduced the expression of PPARγin HSCs cells, thererby promoted the activation of HSCs, with apparent stiffness at 37 kPa. HSCs promoted the expression of ABL in BMSCs at three stiff substrates, but inhibited the expression of alpha-SMA and Col1A1 in BMSCs at 37 kPa, suggesting that co-culture had inhibited the differentiation of BMSCs myofibroblasts, and promoted the differentiation of hepatocyte-like cells, especially at high stiff substrates. In the co-culture of BMSCs and hepatic parenchymal cells, BMSCs had promoted the proliferation of hepatic parenchymal cells at 4.5 kPa. Further, hepatic parenchymal cells had inhibited the expression ofα-SMA in BMSCs, and promoted the expression of Alb, with inhibition of BMSCs differentiation towards myofibroblasts.@*Conclusion@#The differentiation of BMSCs affects the substrate mechanical microenvironment, co-culture of HCs and HSCs. Simultaneously, affecting the function of hepatocytes in relation to the mechanical state of the substrates.

Analysis of clinicopathological features and prognosis of 87 low grade glioma / 中国医师进修杂志

Objective To investigate the clinicopathological features and prognostic factors in patients with low grade glioma. Methods Eighty- seven patients with low grade glioma confirmed by cytological examination were retrospectively analyzed. Results Among 87 patients, male accounted for 62.1%(54/87), and the average age was 36.2 years. The average followed- up time was 51.4 months (3-135 months), and 5 cases were lost in follow-up. The follow- up rate was 94.3% (82/87). The 3- year survival rate was 84.1%, and the 5- year survival rate was 66.7%. The 3- year survival rate was related to clinical symptoms, maximum diameter of tumor and the extent of resection (P<0.01 or<0.05);and the 5- year survival rate was related to years, Karnofsky score, clinical symptoms, maximum diameter of tumor, the pathology classification and extent of resection (P<0.05 or<0.01). Conclusions Low grade glioma is more common in male patients and patients under the age of 40 years. Age below 40 years, Karnofsky score ≥ 70 scores, only epilepsy symptom, tumor diameter below 6.0 cm, oligodendrogliomas, complete resection or subtotal resection are positive prognostic factors. Postoperative radiotherapy can reduce the rate of tumor progression.

Narrative medical enlightenment on medical humanities quality education in China / 中华医学教育探索杂志

Narrative medicine has brought new ideas to medical humanities education, and its con-notation is suitable to Undergraduate Medical Education Standards-Clinical Medicine (Trial). By analyz-ing the proposal and development of narrative medicine, its foundation and core, its programs and methods, as well as its forms and values, the author deepened the understanding of narrative medicine. And through the following four pathways including strengthening the renewal of ideas and concepts on medical humanities quality education, promoting the diversification of teaching ways and methods of medical humanities educa-tion, enhancing the effect of early exposure to clinical medical students, opening the path of integration narrative medicine and evidence-based medicine, the author analyzed narrative medical enlightenment on medical humanities quality education in China.

Secretion and expression of vascular endothelial growth factor and interleukin-8 by SH-SY5Y human neuroblastoma cells / 生物工程学报

To establish vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8) as secretary biomarkers for cell growth on topographic substrates, we have evaluated the secretion and expression of these 2 factors by SH-SY5Y human neuroblastoma cells on poly-L-lactide (PLLA) micropillar arrayed topographic substrates. We fabricated topographic substrates with UV lithography, silicon etching and polydimethylsiloxane-based replica molding, and interfaced SH-SY5Y human neuroblastoma cells with both the topographic substrates and PLLA flat substrates. Cell morphology and spreading were examined with scanning electron microscopy. The secretion and mRNA expression of VEGF and IL-8 were evaluated with enzyme linked immunosorbent assay (ELISA) and real time qPCR, respectively, 24 hours after cell plating. We successfully achieved 4 topographic substrates with a nominal pillar diameter of 2 microm and 4 microm, and a nominal pillar spacing of 2 microm and 7 microm. We found that the secretion and mRNA expression of VEGF and/or IL-8 by SH-SY5Y cells on 2-2 microm (pillar diameter-spacing), 4-2 microm and 4-7 microm topographic substrates were upregulated in comparison to those by cells on PLLA flat substrate, 24 hours after cell plating. Furthermore, both cytokines were even more substantially upregulated on the 2-7 microm substrate than on the other 3 topographic substrates. Compared to those on PLLA flat substrate, cells on topographic substrates showed significant changes in morphology (spreading area, perimeter and roundness), and the increase in the secretion and mRNA expression of VEGF and IL-8 was accompanied with a decrease in cell spreading areas. These results provided evidence that pillar arrayed topography was an important microenvironmental factor in affecting VEGF and IL-8 expression or secretion, and VEGF and IL-8 might serve as important secretary biomarkers for growth on topographic substrates by SH-SY5Y cells.

Fabrication of three-dimensional microwell patterns and their integration with C17. 2 neural stem cells / 生物医学工程学杂志

UV photolithography and hydrofluoric acid wet etching were used to produce silicon master molds and polydimethylsiloxane (PMDS)-based soft lithography was adopted to fabricate three-dimensional poly(lactic-co-glycolic acid) (PLGA) and PDMS microwell patterns with high aspect ratio and channel connection. Nine microwell patterns were thus obtained with different structural dimensions. Patterns were treated with oxygen plasma etching and polylysine coating to enhance hydrophilicity and cell compatibility for subsequent culture of C17. 2 neural stem cells. With proliferation during the culture, C17. 2 cells gradually distributed within the microwells, showing an obviously three-dimensional (3-D) growth behavior. The presence of channel structures greatly favored the 3-D growth of C17. 2 neural stem cells on the microwell patterns. Multi-layered scanning with confocal microscopy and 3-D rendering after carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) staining showed that most C17. 2 cells grew within a range of 30 to 90 microm from the microwell bottom. Immunofluorescence staining indicated that C17. 2 cells within 3-D microwell patterns were uniformly nestin-positive on day 2 after cell plating. It could well be concluded that the microwell patterns thus fabricated were suitable for the 3-D culture and subsequent differentiation of C17. 2 neural stem cells. And the cells can be maintained with uniform stemness properties while cultured in these microwell patterns.

Investigation on culture of rat pulmonary microvascular endothelial cells and their viscoelasticity / 生物医学工程学杂志

It is the infent of this study to establish a simple method for cultivation of rat pulmonary microvascular endothelial cells(PMVECs) and investigate the viscoelasticity of PMVECs. First, we obtained rat's peripheral pulmonary tissue, which then was cut into small pieces and cultured with 3 ml DMEM containing 20% bovine calf serum, 90 U/ml heparin, 4 mmol L-glutamine, 100 U/ml penicillin and 100 micrograms/ml streptomycin. Next, moved away the pulmonary tissue pieces 60 h later, and started passage 2-4 days after continued culture. Last, digested and separated PMVECs and studied viscoelastic coefficients of PMVECs by using micropipette aspiration technique. The results revealed that the cultured PMVECs showed regular cobblestone morphology and conformed with endothelial cells morphological characterization by phase contrast microscopy. PMVECs elastic modulus K1 was 49.3 +/- 9.2 Pa, K2 was 73.2 +/- 24.8 Pa, and it's viscosity factor mu was 19.2 +/- 7.2 Pa. s. These data demonstrate that it is feasible to cultivate PMVECs with tissue pieces method, and PMVECs is of greater rigidity.

The Effects of the Mechanical Stretch on the Adhesion and Growth of Vascular Smooth Muscle Cells in vitro / 生物化学与生物物理进展

An in vitro model was built for researching the effects of strain on vascular smooth muscle cells (VSMCs). The cultured VSMCs was stretched by four-support-bending-beam system, then the project area of cells was measured by computer-image-processing, the adhesion force was measured by micropipette-aspirating system, the α-actin of VSMCs was distinguished by immunocytochemistry and the dynamic of VSMCs was determined by FCM. The results show that: (1) The adhesion force of VSMCs is positively related to time. The adhesion force of unit area is indistinct after stretched for four hour. (2) The amount of α-actin increases with stretching time. (3) The proliferation of VSMCs is a little inhibited by stretched 24 h. These results suggest that the VSMCs in vitro could adjust their behavious to adapt the tension.

Development and Application of New Substrate-stretching Device for Cultured Cells / 医疗卫生装备

Objective To simulate the mechanical environment of cells in vivo and study cellular signal transduction mechanisms. Methods A device was developed which could provide high cell yield, control the time, strain magnitude, direction and frequency of stretch, and applied 10% cyclic strain to cell culture substrate with stretch frequency at 1Hz. Results After being stretched, morphology and cytoskeleton of cells were altered. The major axis of cells and the alignment of stress fibers were vertical to the orientation of cyclic stretch. Conclusion This device provides versatile options for the study on the cellular responses of mechanical loading.