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<p><b>OBJECTIVE</b>To compare the immunomodulatory effects of human amniotic mesenchymal stem cell (hAMSCs) and human bone marrow mesenchymal stem cells (hBMSCs) on peripheral blood T lymphocytes in an in vitro co-culture system.</p><p><b>METHODS</b>hAMSCs and hBMSCs isolated using enzymatic digestion and Ficoll-Hypaque density gradient centrifugation, respectively, were culture-expanded in vitro to obtain the 4th-generation cells. The two MSCs were co-cultured separately with human peripheral blood mononuclear cells stimulated with phytohemagglutinin (PHA-PBMSC) to investigate the changes in T lymphocyte subsets using flow cytomety and the production of interleukin-2 (IL-2) and IL-10 by the T lymphocytes using enzyme-linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>Co-culture with either hAMSCs or hBMSCs significantly increased the proportions of Treg, Th2 and Tc2 and decreased Th1 and Tc1 cell subsets in the PBMCs as compared with the PBMCs cultured alone (P<0.05), and the changes in the PBMCs were similar between the two co-culture systems (P>0.05). In both of the two co-culture systems, IL-2 production by the lymphocytes was significantly lowered (P<0.05) and IL-10 production was significantly increased (P<0.05) as compared with their levels in the PBMCs cultured alone; no significant difference was found in IL-2 or IL-10 levels between the two co-culture systems (P>0.05).</p><p><b>CONCLUSION</b>The MSCs derived from human amnion and bone marrow have similar immunomodulatory effects on the T lymphocytes, suggesting the possibility of using hAMSCs in the treatment of graft-versus-host disease after allogeneic hematopoietic stem cell transplantation.</p>
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Objective To design a virtual reality(VR) system for fracture reduction training using manikin technology.Methods The real-time posture information of the manikin skeleton was acquired by the inertial measurement unit(IMU).3DMAX was used to construct a 3-dimensional human skeleton model based on the human skeleton CT.The model was driven by the received sensor data and displayed the instant attitude.The skeletal attitudes and operation information were displayed in real time through the Visual Studio platform,which was displayed on the screen facing the trainee.Thus,a VA combination interactive training system was constructed.Results and Conclusion A set of simulated fracture reduction training system with high accuracy,low cost and VR combination was designed.Compared with the high-cost,precision optical instrument,the root mean square error (RMSE) of the fraction angle calculation was 0.595° in the static state,and was 1.609° in the dynamic state.This system can provide effective information for training operations and provide a new interactive platform for fracture reduction training.
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Multiple methods of attitude measureinent based on inertial measurement unit (IMU) were summarized.The single sensor's means of attitude measurement and it's relative merits were introduced.What's more,the combined method applied to attitude measurement were highlightedly mentioned.In addition,the ways of attitude measurement with multi-sensors units were listed.It also gave useful recommendations for better selection with regard to specific application.The prospect of IMU applied on human bodies was analysed.
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Novel hydrophobic absorbents were synthesized by immobilizing butyl derivative onto the highly cross-linked agarose beads manufatured in China, which are used as matrix. The effect of the spacer arm length (3C, 8C and 10C) and ligand density (from 13 to 45 micromol/mL) on the hydrophobicity were investigated using purified Hepatitis B surface antigen (HBsAg) expressed by CHO cell lines. Also considering the effects of salt concentration and pH on HBsAg recovery and purification factor, orthogonal experiment design method was used to evaluated the absorbents. The results showed the butyl-S absorbent with the spacer arm length of C8, the ligand density of 22 micromol/mL gel showed the best performance for the separation of HBsAg. Approximately 100% HBsAg recovery and 60 as purification fold were achieved by this media under the operating condition of pH 7.0 and 9% of salt concentrateion.
Subject(s)
Animals , Cricetinae , Adsorption , CHO Cells , Chromatography, Agarose , Methods , Cricetulus , Hepatitis B Surface Antigens , Hydrophobic and Hydrophilic Interactions , Recombinant ProteinsABSTRACT
A new hydrophobic absorbent based on homemade highly cross-linked agarose beads was synthesized by immobilizing butyl derivative onto the matrix linkage. The density of ligand was controlled by adjusting the concentration of butanethiol and the synthesis route was optimized by evaluating the purification efficiency of recombinant Hepatitis B surface antigen (HBsAg) expressed by Chinese hamster ovary (CHO) cell line. A high performance absorbent was finally screened out with up to 80% of HBsAg recovery and purification-fold (PF) about 20. Furthermore, the column pressure was about 0.06 MPa under the flow rate of 500cm/h, and no leaked butyl were detected after exposing the gel in common buffers, chaotropic agents, high concentrations of denaturing agents such as guanidine hydrochloride, urea and polar organic solvents. These results demonstrated that the absorbent have high physico-chemical stability, so it was available for the downstream process. Finally, after scaled up to 2L wet gel/batch, the absorbent was applied to the integration of three-step chromatography and obtained the purified CHO-HBsAg with 95% purity by SDS-PAGE and HPLC, which meet the requirements of SFDA. The purification efficiency and the reproducible ability of the absorbents were also evaluated from batch-to-batch. The results demonstrated that the absorbent met the requirement of scalable, reproducible, economic effect as well. This absorbent is a promising alternative exported HIC gel for wildly being used in Chinese pharmaceutical industries.
Subject(s)
Animals , Cricetinae , Humans , CHO Cells , Chromatography, Agarose , Methods , Cricetulus , Hepatitis B Surface Antigens , Genetics , Recombinant Proteins , GeneticsABSTRACT
Objective To evaluate the metabolic characteristics of insulin secretion and insulin sensitivity in isolated postchallenge hyperglycemia(IPH) and to clarify the factors responsible for the development of IPH. Methods(Eight hundred) and fifty subjects were classified into the following three groups based on the results of a 75-g oral glucose tolerance test(OGTT): normal glucose tolerance(NGT),n=557;isolated impaired glucose tolerance(iIGT),n=146;and IPH,n=147.Insulin secretion(insulinogenic index) and insulin sensitivity(insulin sensitivity index) were identified in the three groups. Results From NGT to iIGT and IPH in these subjects,the insulinogenic index and insulin sensitivity index were gradually decreased(P
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The dissociation of virus-like particles of Hepatitis B surface antigen (HBsAg) during the adsorption-desorption on the solid-phase of chromatography is a main challenge for its purification. Herein, poly (ethylene glycol) (PEG) was applied as an additive during the purification of HBsAg from recombinant Chinese hamster ovary (CHO) cell culture to improve the HBsAg recovery and protect its structural assembly. The presence of 1% of PEG10000 in the mobile phase of ion-exchange chromatography (IEC) of DEAE-Sepharose FF column could increase the recovery of HBsAg from about 55% to 80%, with a similar purification (-fold) (about 12) compared with the absence of PEG. Importantly, glycosylated protein forms of HBsAg were reserved well by PEG-accompanied chromatography. Furthermore, size exclusion chromatography-multiangle laser light scattering (SEC-MALLS) analysis was performed on line to monitor the aggregates, particle size and molecular weight distribution of HBsAg. The results demonstrated that the HBsAg particle size and assembly are more homogenous after adding PEG in the mobile phase of IEC than no PEG added in the mobile phase.
Subject(s)
Animals , Cricetinae , Humans , CHO Cells , Chromatography, Ion Exchange , Methods , Cricetulus , Hepatitis B Surface Antigens , Genetics , Polyethylene Glycols , Chemistry , Recombinant Proteins , GeneticsABSTRACT
<p><b>AIM</b>To explore the effect of ligustrazine injection on the expression of heme oxygenase-1 (HO-1) in rabbits with pulmonary ischemia/reperfusion injury after.</p><p><b>METHODS</b>Single lung ischemia/reperfusion injury animal model was used in vivo. Twenty rabbits were randomly divided into two groups( n = 10, in each), pulmonary ischemia/reperfusion injury (I/R) group and I/R + ligustrazine injection (LGT) group. The tissue slides were stained by immunohistochemistry (IHC) and in situ hybridization (ISH) for HO-1 to detect the expression of HO-1 in lung and to analyze the absorbance, wet to dry ratio of lung tissue weight (W/D) and the injured alveoli rate (IAR) were measured at 180 minutes after lung reperfusion. Meanwhile the lung tissue slide was prepared for electron microscopic observation at 180 minutes after reperfusion.</p><p><b>RESULTS</b>HO-1 expression was upregulated in two groups in the pulmonary endothelial cells, part of pulmonary vascular smooth muscle cells, extima of vessels and epithelial cells of airway, the absorbance was 0.168 +/- 0.016 (0.148 +/- 0.013), 0.186 +/- 0.014 (0.158 +/- 0.012) respectively.The LGTI group showed higher absorbance than those of the I/R group (P < 0.01), lower W/D and IAR values than those of the I/R group (P < 0.01) significantly and lighter abnormal changes of the lung tissue in morphology than those of the I/R group.</p><p><b>CONCLUSION</b>Ligustrazine injection possesses notable protective effects on I/R in rabbits by increasing the expression of HO-1 in lung.</p>