1.Experimental study on the transplantation of neural stem cells derived from bone marrow of crab-eating macaque
Yiquan KE ; Gang LI ; Ruxiang XU ; Xiaodan JIANG ; Wenping CHENG
Chinese Journal of Tissue Engineering Research 2005;9(5):200-201
BACKGROUND: Researches indicated that bone marrow stem cells (BMSCs) could differentiated into neural stem cells in vitro, but what was the role of neural stem cells(NSCs) in the recovery of cortical injury,whether the NSC is capable of growing and migration in injured still remained unknown.OBJECTIVE: To explore the growing state of autograft NSC derived from crab-eating macaque BMSC transplanted in brain.DESIGN: Prospective case control study based on experimental animals.SETTING: Department of neurosurgery in a hospital of a military medical university.MATERIALS: This study was carried out at Center Laboratory of Neurological Research Institute, Zhujiang Hospital affiliated to the First Military Medical University of Chinese PLA. Six healthy adult crab-eating macaques were purchased from the South China Primate Animal Center.INTERVENTIONS: BMSCs harvested from six crab-eating macaques were cultured in vitro and induced to differentiate into neural stem cells, which then labeled by bromodeoxyuridine(BrdU) and autografted into brains.MAIN OUTCOME MEASURES: Brain tissues underwent hematoxylin and eosin(HE) staining and immunohistochemical staining before observed under optical microscope.RESULTS: The results of HE staining showed that the cell number in injured brain vas obviously higher in both instant and delayed transplanting groups than sham-transplanting group; moreover cells were proved reacting to BrdU by immunohistochemical staining in cortical injuries of both groups at 1-6 months following stem cells autograft, as well as at neighboring white matters at half year later, but no BrdU positive cells could be found in traumatic controls, sham-transplanting group and normal brains.CONCLUSION: NSCs derived from in vitro cultured BMSCs were proved capable of surviving, proliferating, differentiating and migrating in cortex after autograft, so that BMSCs is considered as replacing cells or the source of NSCs; moreover autograft stem cells could survive, proliferate and migrate in old cortical traumatic focus.
2.Wortmannin inhibits K562 lukemic cells by regulating PI3k/Akt channel in vitro.
Qing, WU ; Yan, CHEN ; Guohui, CUI ; Yiquan, CHENG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2009;29(4):451-6
The inhibitory effect of wortmannin on leukemic cells and the possible mechanisms were examined. K562 cells were treated with wortmannin of various concentrations (3.125-100 nmol/L) for 0-72 h. MTT assay was used to evaluate the inhibitory effect of wortmannin on the growth of K562 cells. Cell apoptosis was detected by both Annexin-V FITC/PI double-labeled cytometry and transmission electron microscopy (TEM). The expression of p-Akt, T-p-Akt, NF-kappaBp65 and IKK-kappaB was determined by Western blotting and reverse transcription-polymerase chain reaction (RT-PCR). Our results showed that wortmannin obviously inhibited growth and induced apoptosis of K562 cells in vitro in a time- and dose-dependent manner. The IC(50) value of wortmannin for 24 h was 25+/-0.14 nmol/L. Moreover, wortmannin induced K562 cells apoptosis in a dose-dependent manner. TEM revealed typical morphological changes of apoptosis in wortmannin-treated K562 cells, such as chromatin condensation, karyopyknosis, karyorhexis and apoptotic bodies. Additionally, several important intracellular protein kinases such as p-Akt, NF-kappaBp65 and IKK-kappaB experienced degradation of various degrees in a dose-dependent manner both at protein level and transcription level when cultured with wortmannin, but the expression of total Akt showed no change. It is concluded that wortmannin can inhibit the proliferation and induce apoptosis of K562 leukemia cells possibly by down-regulating the survival signaling pathways (PI3K/Akt and NF-kappaB channels).
3.Analysis of surgical treatment of 205 cases of spinal fracture
Chunhua TAN ; Zhenhe GUO ; Daohai LU ; Zhe WEN ; Yiquan CHENG ; Kun HE
Journal of Chinese Physician 2011;(z1):32-35
Objective To discuss the surgical treatment of spinal injury, and provide insights on key points and related issues for operations. Methods Two hundred and five cases of spinal fracture were treated through posterior surgical treatment. Under C-arm X-ray monitoring, surgeries had been operated on pedicle screws insertion, vertebral canal decompression, over-extending reduction position, and placed the connecting rod, knocked in the bone graft and finally transplanted the paraspinal bone. Results After operations , the height and morphology of vertebral bodies and spinal physiological curvature were basically recovered analyzed by X ray examination. The follow-up results (in the average of 14 to 36 months) indicated that there were 4 cases of delayed infection, 7 cases of loosen screw, 6 cases of broken screw (14 screws)and 1 case of broken stick, with no secondary nerve injury or other syndromes. Conclusion The vertebral pedicle screw internal fixation manipulated easily, which could sufficiently enlarge vertebral canal in order to decompression. In addition, during the operation, together with over-extending reduction position is beneficial to regain the height of fractured vertebral bodies.
4.Differentiative capability of bone marrow mesenchymal cells into neural stem cells in cats
Yiquan KE ; Wenping CHENG ; Gang LI ; Xiaodan JIANG ; Ruxiang XU ; Songqing WANG
Chinese Journal of Tissue Engineering Research 2005;9(22):233-235
BACKGROUND: Bone marrow mesenchymal cells, multiple-potential non-hematopoiefic stem cells adhering to the wall in vitro culture, can be induced to proliferate and differentiate towards neurons and glia cells.OBJECTIVE: To investigate the growth state of cat bone marrow mesenchymal cells in vitro culture, as well as the capability to differentiate towards neural stem cells.DESIGN: A randomized sampling study.SETTING: Department of Neurosurgery, Zhujiang Hospital, Southern Medical University.MATERIALS: This study was carried out at the Central Laboratory of the General Military Neurological Research Institute, Zhujiang Hospital, Southern Medical University between January and December 2002. Twenty healthy home-raised cats, aged 1.0 - 2.0 years and weighing 2. 5 - 4.0 kg, male and female in half, were provided by the Animal Center of the First Military Medical University of Chinese PLA.INTERVENTIONS: Bone marrows were randomly aspirated from the left or right hindlimbs in order to separate bone marrow mesenchymal cells, then the bone marrow mesenchymal cells single cell suspension was co-cultured with neural stem cell culture media in vitro so as to induce differentiation to neural stem cells with tretinoin. CK2 type inverted optical microscope(Olympus,Japan) was used to observe the growth of bone marrow mesenchymal cells in vitro culture, as well as 4, 12, 24, 48 hours of induction upon eliminating or not eliminating the wall-adhering cells. Bone marrow mesenchymal cells in stem cell stage were identified under Olympus optical microscope with modified immunohistochemical staining.MAIN OUTCOME MEASURES: The growth state and the immunocytochemical staining of living bone marrow mesenchymal cells exposed to experimental intervention were observed under the Olympus inverted optical microscope.RESULTS: Data from the 20 cats were analyzed without loss. Reversed microscopic observation revealed that cat bone marrow mesenchymal cells becrame larger when cultured in vitro, which were rich in plasmic granules with prominence projecting, adhering to the wall and forming cell clones. These cells were then successively cultured, and imnunohistochemical staining analysis suggested that the passaged bone marrow mesenchymal cells could express neural stem cells-specific antigen Nestin and differentiate towards glia-like cells and neuron-like cells.CONCLUSION: Cat bone marrow mesenchymal cells possess the characteristics of stem cells; they can be amplified into cell clones and induced to express the property of neural glia cells and neuron-like cells under proper condition.
5.A pilot study on the culture and differentiation of bone marrow stromal cells from SD rats.
Gang LI ; Yiquan KE ; Xiaodan JIANG ; Ruxiang XU ; Yuxi ZHOU ; Wei WANG ; Wenping CHENG ; Keli LIAO
Journal of Biomedical Engineering 2004;21(1):16-20
In order to observe the growth, expansion and differentiation of the cultured bone marrow stromal cells (BMSC), we isolated the BMSC from adult SD rats and cultivated them with LIF and bFGF. Then, we cultured and induced the stem cells by using retinoic acid and the culture medium confected in our lab by ourselves. We found that the BMSC could expand and generate clones when they were cultured in vitro. These cells subcultured grew rapidly and differentiated into neuron-like cells and astrocyte-like cells. The results showed that BMSC have the abilities to self renew and differentiate, thus demonstrating the culture method we used is suitable for the culture of BMSC in vitro. The bone marrow stromal cell is not difficult to obtain; it is capable of expanding and differentiating in culture. If the culture condition is appropriate, it can differentiate into neuron and astrocyte. So, it is a kind of perfect seed cells.
Actihaemyl
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pharmacology
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Animals
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Bone Marrow Cells
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cytology
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drug effects
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Cell Culture Techniques
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Cell Differentiation
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drug effects
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Cell Proliferation
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drug effects
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Cells, Cultured
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Fibroblast Growth Factor 2
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pharmacology
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Neurons
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cytology
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Pilot Projects
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Rats
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Rats, Sprague-Dawley
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Stem Cells
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cytology
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Stromal Cells
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cytology
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drug effects
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Tretinoin
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pharmacology
6.Wortmannin Inhibits K562 Lukemic Cells by Regulating PI3k/Akt Channel In Vitro
WU QING ; CHEN YAN ; CUI GUOHUI ; CHENG YIQUAN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2009;29(4):451-456
ssion of total Akt showed no change. It is concluded that wortmannin can inhibit the proliferation and induce apoptosis of K562 leukemia cells possibly by down-regulating the survival signaling pathways (PI3K/Akt and NF-κB channels).
7.Diabetic retinopathy fundus image generation based on generative adversarial networks
Cheng WAN ; Peng ZHOU ; Luhui WU ; Yiquan WU ; Jianxin SHEN ; Hui YE
Chinese Journal of Experimental Ophthalmology 2019;37(8):613-618
Objective To generate various types of diabetic retinopathy ( DR) fundus images automatically by computer vision algorithm. Methods A method based on deep learning to generate fundus images was proposed,which used the vascular vein of the fundus image and the text description of lesions as the constraint conditions to generate fundus image. The text description was encoded by using a long short-term memory ( LSTM) , and the vascular vein image was encoded by a convolutional neural network (CNN). Then the encoded information was combined and used to generate a fundus image by generative adversarial networks ( GAN ) . Results The results showed that the algorithm can generate realistic fundus images. However, the image detail features were not obvious because the text-encoded recurrent neural network ( RNN ) loss function did not converge well. Conclusions Using the GAN can generate realistic DR fundus images, which has certain application value in expanding medical data. However,the generation of detail features in small areas still needs improvement.