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Lu P.,CAS Shanghai Institutes for Biological Sciences | Lu P.,Zhejiang University | Chen J.,CAS Shanghai Institutes for Biological Sciences | Chen J.,Zhejiang University | And 12 more authors.
Stem Cell Reviews and Reports | Year: 2015

Immune rejection hinders the application of human embryonic stem cells (hESCs) in transplantation therapy. Human leukocyte antigens (HLAs) on the cell surface are the major cause of graft rejection. In this study, we generatedHLA class I-deficient hESCs via disruption of beta 2-microglobulin (β2m), the light chain of HLA Class I. We found that HLA class I proteins were not present on the cell surface of β2mnull hESCs. These cells showed the same pluripotency as wildtype hESCs and demonstrated hypoimmunogenicity. Thus, HLA class I-deficient hESCs might serve as an unlimited cell source for the generation of universally compatible “off-the-shelf” cell grafts, tissues or organs in the future. © Springer Science+Business Media New York 2013


Cheng J.,Central South University | Cheng J.,Xiangtan Center Hospital | Fan C.,Central South University | Tang M.,Central South University | And 2 more authors.
Cardiology (Switzerland) | Year: 2016

Objectives: Tetralogy of Fallot (TOF) with major aortopulmonary collaterals (MAPCA) is a well-known but always severe congenital heart disease. This study was designed to explore proper management after radical correction of TOF with MAPCA based on a hierarchical approach. Methods: The following data were collected from 39 patients planned to undergo radical correction of TOF: age, weight, number of aortopulmonary collaterals, total lumen diameter and collateral diameter-to-body weight ratio, transcatheter occlusion and cardiac catheterization findings, mechanical ventilation time, and ICU monitoring time. The patients were divided into 4 groups by collateral diameter-to-body weight ratio as follows: <0.200 mm/kg (group 1), 0.200-0.500 mm/kg (group 2), >0.500 mm/kg (group 3), and no MAPCA (group 4). Data analysis was performed using IBM SPSS Statistics software for Mac version 22.0 (SPSS Inc., Chicago, Ill., USA) with logistic regression and Fisher's exact test. Results: Most of the patients recovered well after radical correction; postoperative complications occurred in 12 patients and included bloody sputum, low cardiac output syndrome, and severe pulmonary infection that led to tracheotomy. By prolonging the mechanical ventilation time of the patients with postoperative complications, the conditions in 3 patients were improved. However, in the remaining patients, the condition worsened until transcatheter occlusions were performed. Transcatheter occlusion was performed in all 7 patients in group 3 (100%). Only 2 of the 8 patients in group 2 required transcatheter occlusion (25%), and none of the 9 patients in group 1 required transcatheter occlusion (0%). Only 1 patient (group 3) died after radical correction. The transcatheter occlusion results showed a strong association with the total lumen diameter and the collateral diameter-to-body weight ratio (p < 0.05) but no obvious association with age, weight, or the number of aortopulmonary collaterals (p > 0.05). Conclusions: Postoperative management of patients with TOF and MAPCA has great significance. To reduce the morbidity and mortality, transcatheter coil embolization or surgical ligation should be performed in patients with a collateral diameter-to-body weight ratio of at least 0.500 mm/kg. In patients with values between approximately 0.200 and 0.500 mm/kg, prolongation of mechanical ventilation should have priority over transcatheter occlusion, and for patients with values below 0.200 mm/kg no additional treatment is needed. © 2016 S. Karger AG, Basel.


Cui D.,Zhejiang University | Wang J.,Zhejiang University | Zeng Y.,Zhejiang University | Rao L.,CAS Shanghai Institutes for Biological Sciences | And 6 more authors.
Bioscience, Biotechnology and Biochemistry | Year: 2016

Human embryonic stem cells (hESCs) are thought to be a promising resource for cell therapy, while it has to face the major problem of graft immunological rejection. Major histocompatibility complex (MHC) class I expressed on the cell surface is the major cause of graft rejection. Transporter associated with antigen presentation 1 (TAP1) and TAPassociated glycoprotein (TAPBP) play important roles in regulating MHC class I expression. In this study, we generated TAP1- and TAPBP-deficient hESC lines, respectively, using transcription activator-like effector nucleases technique. These cells showed deficient expression of MHC class I on the cell surface and reduced immunogenicity compared with wild types, but maintained normal pluripotency, karyotypes, and differentiation ability. Thus, our findings are instrumental in developing a universal cell resource with both pluripotency and hypo-immunogenicity for transplantation therapy in the future. © 2016 Japan Society for Bioscience, Biotechnology, and Agrochemistry.


Chen G.,PhysioSign Laboratory | Yao L.,PhysioSign Laboratory | Zhao R.,Baotou Central Hospital | Zeng J.,Xiangtan Center Hospital | Liu M.,Wuhan Asia Heart HospitalHubei
International Journal of Cardiology | Year: 2016

Since ECG was invented in 1903, this is the first time in history that a full information multi-band and multi-linear electrophysiological cardiogram has been used to successfully scan and record on the human body surface. Since it is able to record various multi-band, multi-track linear electric signals of cardiac electrophysiological activities that correspond to different regions of the entire heart, it has thus been denominated as “electrophysiocardiogram” (EPCG). A traditional ECG is always represented by a characteristic wave form, which resembles a string. For a long period of time, ECG has had a lot of mysteries surrounding it, it maybe because ECG has a lot of mixed signals buried in such convolutionary forms, which limits the amount of the signals that are discernable and determinable. For the first time, the EPCG technology has allowed cardiac signals to be convoluted into the linear wave form, which is then processed through various new approaches featuring multiple frequency bands, multiple dimensions and multiple patterns, and consequentially recorded as the following types of signals within the ranges of P wave and T wave: multiple frequency band signals, signals of different regions and different locations, forward waves and negative waves. Therefore, EPCG may help to solve many puzzling scientific questions regarding heart, such as exactly how many electric signals are involved in heart excitation, pacing, conduction and action, as well as many other intriguing questions about heart, and thus would become a very helpful tool in clinical practice. © 2016


Wu Z.,CAS Shanghai Institutes for Biological Sciences | Li H.,Xiangtan Center Hospital | Rao L.,CAS Shanghai Institutes for Biological Sciences | He L.,CAS Shanghai Institutes for Biological Sciences | And 10 more authors.
Journal of Genetics and Genomics | Year: 2011

Human embryonic stem cells (hESCs) can self-renew indefinitely and differentiate into all cell types in the human body. Therefore, they are valuable in regenerative medicine, human developmental biology and drug discovery. A number of hESC lines have been derived from the Chinese population, but limited of them are available for research purposes. Here we report the derivation and characterization of two hESC lines derived from human blastocysts of Chinese origin. These hESCs express alkaline phosphatase and hESC-specific markers, including Oct4, Nanog, SSEA-3, SSEA-4, TRA-1-60 and TRA-1-81. They also have high levels of telomerase activity and normal karyotypes. These cells can form embryoid body in vitro and can be differentiated into all three germ layers in vivo by teratoma formation. The newly established hESCs will be distributed for research purposes. The availability of hESC lines from the Chinese population will facilitate studies on the differences in hESCs from different ethnic groups. © 2011.


Bao L.,CAS Shanghai Institutes for Biological Sciences | He L.,CAS Shanghai Institutes for Biological Sciences | Chen J.,CAS Shanghai Institutes for Biological Sciences | Wu Z.,CAS Shanghai Institutes for Biological Sciences | And 15 more authors.
Cell Research | Year: 2011

Reprogramming of somatic cells in the enucleated egg made Dolly, the sheep, the first successfully cloned mammal in 1996. However, the mechanism of sheep somatic cell reprogramming has not yet been addressed. Moreover, sheep embryonic stem (ES) cells are still not available, which limits the generation of precise gene-modified sheep. In this study, we report that sheep somatic cells can be directly reprogrammed to induced pluripotent stem (iPS) cells using defined factors (Oct4, Sox2, c-Myc, Klf4, Nanog, Lin28, SV40 large T and hTERT). Our observations indicated that somatic cells from sheep are more difficult to reprogram than somatic cells from other species, in which iPS cells have been reported. We demonstrated that sheep iPS cells express ES cell markers, including alkaline phosphatase, Oct4, Nanog, Sox2, Rex1, stage-specific embryonic antigen-1, TRA-1-60, TRA-1-81 and E-cadherin. Sheep iPS cells exhibited normal karyotypes and were able to differentiate into all three germ layers both in vitro and in teratomas. Our study may help to reveal the mechanism of somatic cell reprogramming in sheep and provide a platform to explore the culture conditions for sheep ES cells. Moreover, sheep iPS cells may be directly used to generate precise gene-modified sheep. © 2011 IBCB, SIBS, CAS All rights reserved.


PubMed | Jiangnan University, Xiangtan Center Hospital and Zhejiang University
Type: | Journal: Biological research | Year: 2015

Theoretically human embryonic stem cells (hESCs) have the capacity to self-renew and differentiate into all human cell types. Therefore, the greatest promise of hESCs-based therapy is to replace the damaged tissues of patients suffering from traumatic or degenerative diseases by the exact same type of cells derived from hESCs. Allograft immune rejection is one of the obstacles for hESCs-based clinical applications. Human leukocyte antigen (HLA) II leads to CD4(+) T cells-mediated allograft rejection. Hence, we focus on optimizing hESCs for clinic application through gene modification.Transcription activator-like effector nucleases (TALENs) were used to target MHC class II transactivator (CIITA) in hESCs efficiently. CIITA (-/-) hESCs did not show any difference in the differentiation potential and self-renewal capacity. Dendritic cells (DCs) derived from CIITA (-/-) hESCs expressed CD83 and CD86 but without the constitutive HLA II. Fibroblasts derived from CIITA (-/-) hESCs were powerless in IFN- inducible expression of HLA II.We generated HLA II defected hESCs via deleting CIITA, a master regulator of constitutive and IFN- inducible expression of HLA II genes. CIITA (-/-) hESCs can differentiate into tissue cells with non-HLA II expression. Its promising that CIITA (-/-) hESCs-derived cells could be used in cell therapy (e.g., T cells and DCs) and escape the attack of receptors CD4(+) T cells, which are the main effector cells of cellular immunity in allograft.


Wang F.,Central South University | Yang Y.,Xiangtan Center Hospital | Feng Q.,Central South University | Bu G.,Central South University | And 5 more authors.
Journal of Central South University (Medical Sciences) | Year: 2012

Objective: To construct the RNAi targeting tumor necrosis factor receptor associated factor (TRAF1) gene, and to explore the effect of interference targeting TRAF1 on the biological behavior of gastric cancer cells. Methods: We detected the expression of TRAFl in BGC823, SGC7901, and MGC803 gastric cancer cell lines through the real-time PCR and Western blot; then we constructed three pLVX-shRNA-TRAFl-shRNAs expression vector targeting TRAFl. When TRAFl was interfered successfully, we selected the strongest interference efficiency ShRNA by real-time PCR and Western blot. Based on interference targeting TRAFl on gastric cancer, we tested the cell proliferation activity and apoptosis through MTT assay and flow cytometry, and the cell migration by transwell migration assay. Results: The expression of TRAF1 was increased in BGC823, SGC7901, and MGC803 gastric cancer cell lines compared with gastric epithelial cells (P<0.05), and the highest expression was in BGC823 gastric cell line. In the three TRAF1 shRNAs, the strongest interference efficiency shRNA was pLVX-shRNA-TRAFl-shRNA2. When the gene TRAF1 of BGC823 was interfered, the cell growing power was weakened and the apoptosis rate increased, and the cell migration had no difference. Conclusion: The expression of TRAF1 is up-regulated in gastric cancer cell lines BGC823, SGC7901, and MGC803, and the most obvious one is BGC823. The interference targeting TRAF1 can successfully inhibit the expression of TRAF1 in gastric cancer cell line BGC823. TRAF1 can inhibit the apoptosis of BGC823 cells.

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