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Thymosin Beta-4 Recombinant Adeno-associated Virus Enhances Human Nucleus Pulposus Cell Proliferation and Reduces Cell Apoptosis and Senescence

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Chinese Medical Journal 2015年第11期128卷 1529-1535页

作者： Yuan-Yi Wang Qing-san Zhu Yi-Wei Wang Ruo-Feng Yin Department of Spine Surgery The First Hospital of Jilin University Changchun Jilin 130021 China Department of OrthoPedics China-Japan UnionHospital of Jilin University C-hangchun Jilin 130021 China Department of Pathophysiology Jiangsu Jiankang Vocational College Nanjing Jiangsu 210029 China

Background： Thymosin beta-4 （TB-4） is considered key roles in tissue development, maintenance and pathological processes. The study aimed to prove TB-4 positive biological function on nucleus pulposus （NP） cell apoptosis and slowing the process of cell aging while increasing the cell proliferation. Methods： TB-4 recombinant adeno-associated virus （AAV） was constructed and induced to human NP cells. Cell of same group were cultured without gene modification as controlled group. Proliferation capacity and cell apoptosis were observed during 6 passages of the cells. Morphology and expression of the TB-4 gene were documented as parameter of cell activity during cell passage. Results： NP cells with TB-4 transfection has normal TB-4 expression and exocytosis. NP cells with TB-4 transfection performed significantly higher cell activity than that at the control group in each generation. TB-4 recombinant AAV-transfected human NP cells also show slower cell aging, lower cell apoptosis and higher cell proliferation than control group. Conclusions： TB-4 can prevent NP cell apoptosis, slow NP cell aging and promote NP cell proliferation. AAV transfection technique was able to highly and stably express TB-4 in human NP cells, which may provide a new pathway for innovation in the treatment of intervertebral disc degenerative diseases.

关键词： Biological Treatment Cell Aging Degeneration of Intervertebral Disc Thymosin Beta-4

Transplantation of erythropoietin gene-modified neural stem cells improves the repair of injured spinal cord

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Neural Regeneration Research 2015年第9期10卷 1483-1490页

作者： Min-fei Wu Shu-quan Zhang Rui Gu Jia-bei Liu Ye Li Qing-san Zhu Department of Orthopedics the Second Hospital of Jilin University Department of Orthopedics Tianjin Nankai Hospital Department of Orthopedics China-Japan Union Hospital of Jilin University

The protective effects of erythropoietin on spinal Here, the eukaryotic expression plasmid pcDNA3.1 cord injury have not been well described. human erythropoietin was transfected into rat neural stem cells cultured in vitro. A rat model of spinal cord injury was established using a free falling object. In the human erythropoietin-neural stem cells group, transfected neural stem cells were injected into the rat subarachnoid cavity, while the neural stem cells group was inject- ed with non-transfected neural stem cells. Dulbecco＇s modified Eagle＇s medium/F12 medium was injected into the rats in the spinal cord injury group as a control. At 1-4 weeks post injury, the motor function in the rat lower limbs was best in the human erythropoietin-neural stem ceils group, followed by the neural stem cells group, and lastly the spinal cord injury group. At 72 hours, compared with the spinal cord injury group, the apoptotic index and Caspase-3 gene and protein expressions were apparently decreased, and the bd-2 gene and protein expressions were noticeably increased, in the tissues surrounding the injured region in the human erythro- poietin-neural stem cells group. At 4 weeks, the somatosensory evoked potential latencies were cavities were clearly smaller and the motor and remarkably shorter in the human erythropoi- etin-neural stem cells group and neural stem cells group than those in the spinal cord injury group. These differences were particularly obvious in the human erythropoietin-neural stem cells group. More CM-Dil-positive cells and horseradish peroxidase-positive nerve fibers and larger amplitude motor and somatosensory evoked potentials were found in the human erythro- poietin-neural stem cells group and neural stem cells group than in the spinal cord injury group. Again, these differences were particularly obvious in the human erythropoietin-neural stem cells group. These data indicate that transplantation of erythropoietin gene-modified neural stem cells into the subarachnoid cavity to help repair spinal cord inj

关键词： nerve regeneration spinal cord injury neural stem cells erythropoietin motor function subarachnoid cavity transplantation injury recovery neural regeneration

Transplantation of human telomerase reverse transcriptase gene-transfected Schwann cells for repairing spinal cord injury

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Neural Regeneration Research 2015年第12期10卷 2040-2047页

作者： Shu-quan Zhang Min-fei Wu Jia-bei Liu Ye Li Qing-san Zhu Rui Gu Department of Orthopedics Tianjin Nankai Hospital Department of Spine Surgery Orthopedic HospitalSecond HospitalClinical HospitalJilin University Department of Orthopedics China-Japan Union HospitalJilin University

Transfection of the human telomerase reverse transcriptase（h TERT）gene has been shown to increase cell proliferation and enhance tissue repair.In the present study,h TERT was transfected into rat Schwann cells.A rat model of acute spinal cord injury was established by the modified free-falling method.Retrovirus PLXSN was injected at the site of spinal cord injury as a vector to mediate h TERT gene-transfected Schwann cells（1×10^（10）/L;10μL）or Schwann cells（1×10^（10）/L;10μL）without h TERT gene transfection.Between 1 and 4 weeks after model establishment,motor function of the lower limb improved in the h TERT-transfected group compared with the group with non-transfected Schwann cells.Terminal deoxynucleotidyl transferase-mediated d UTP nick-end labeling and reverse transcription-polymerase chain reaction results revealed that the number of apoptotic cells,and gene expression of aquaporin 4/9 and matrix metalloproteinase 9/2decreased at the site of injury in both groups;however,the effect improved in the h TERT-transfected group compared with the Schwann cells without h TERT transfection group.Hematoxylin and eosin staining,PKH26 fluorescent labeling,and electrophysiological testing demonstrated that compared with the non-transfected group,spinal cord cavity and motor and sensory evoked potential latencies were reduced,while the number of PKH26-positive cells and the motor and sensory evoked potential amplitude increased at the site of injury in the h TERT-transfected group.These findings suggest that transplantation of h TERT gene-transfected Schwann cells repairs the structure and function of the injured spinal cord.

关键词： nerve regeneration spinal cord injury Schwann cells transplantation motor function telomerase reverse transcriptase proliferation modification cells neural regeneration

Neuroprotective effects of electroacupuncture on early- and late-stage spinal cord injury

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Neural Regeneration Research 2015年第10期10卷 1628-1634页

作者： Min-fei Wu Shu-quan Zhang Jia-bei Liu Ye Li Qing-san Zhu Rui Gu Department of Orthopedics Bethune Second Hospital of Jilin University Department of Orthopedics Tianjin Nankai Hospital Department of Orthopedics China-Japan Union Hospital Jilin University

Previous studies have shown that the neurite growth inhibitor Nogo-A can cause secondary neural damage by activating Rho A. In the present study, we hypothesized that electroacupuncture promotes neurological functional recovery after spinal cord injury by inhibiting Rho A expression. We established a rat model of acute spinal cord injury using a modification of Allen＇s method. The rats were given electroacupuncture treatment at Dazhui（Du14）, Mingmen（Du4）, sanyinjiao（SP6）, Huantiao（GB30）, Zusanli（ST36） and Kunlun（BL60） acupoints with a sparsedense wave at a frequency of 4 Hz for 30 minutes, once a day, for a total of 7 days. Seven days after injury, the Basso, Beattie and Bresnahan（BBB） locomotor scale and inclined plane test scores were significantly increased, the number of apoptotic cells in the spinal cord tissue was significantly reduced, and Rho A and Nogo-A m RNA and protein expression levels were decreased in rats given electroacupuncture compared with rats not given electroacupuncture. Four weeks after injury, pathological tissue damage in the spinal cord at the site of injury was alleviated, the numbers of glial fibrillary acidic protein- and neurofilament 200-positive fibers were increased, the latencies of somatosensory-evoked and motor-evoked potentials were shortened, and their amplitudes were increased in rats given electroacupuncture. These findings suggest that electroacupuncture treatment reduces neuronal apoptosis and decreases Rho A and Nogo-A m RNA and protein expression at the site of spinal cord injury, thereby promoting tissue repair and neurological functional recovery.

关键词： nerve regeneration spinal cord injury electroacupuncture locomotion Rho A Nogo-A glial fibrillary acidic protein neurofilament 200 neural regeneration

Senegenin inhibits neuronal apoptosis after spinal cord contusion injury

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Neural Regeneration Research 2016年第4期11卷 657-663页

作者： Shu-quan Zhang Min-fei Wu Rui Gu Jia-bei Liu Ye Li Qing-san Zhu Jin-lan Jiang Department of Orthopedics Nankai Hospital Department of Orthopedics the Second Hospital of Jilin University Department of Orthopedics China-Japan Union HospitalJilin University Scientific Research Center China-Japan Union HospitalJilin University

Senegenin has been shown to inhibit neuronal apoptosis,thereby exerting a neuroprotective effect.In the present study,we established a rat model of spinal cord contusion injury using the modified Allen＇s method.Three hours after injury,senegenin（30 mg/g） was injected into the tail vein for 3 consecutive days.Senegenin reduced the size of syringomyelic cavities,and it substantially reduced the number of apoptotic cells in the spinal cord.At the site of injury,Bax and Caspase-3 m RNA and protein levels were decreased by senegenin,while Bcl-2 m RNA and protein levels were increased.Nerve fiber density was increased in the spinal cord proximal to the brain,and hindlimb motor function and electrophysiological properties of rat hindlimb were improved.Taken together,our results suggest that senegenin exerts a neuroprotective effect by suppressing neuronal apoptosis at the site of spinal cord injury.

关键词： nerve regeneration spinal cord contusion senegenin thinleaf milkwort root motor function apoptosis electrophysiology neural regeneration

Effect of Quercetin on Atherosclerosis Based on Expressions of ABCA1, LXR-α and PCSK9 in ApoE-/- Mice

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Chinese Journal of Integrative Medicine 2020年第2期26卷 114-121页

作者： LI Shan-shan CAO Hui SHEN Ding-zhu CHEN Chuan XING san-li DOU Fang-fang JIA qing-ling Shanghai Geriatric Institute of Chinese Medicine Shanghai University of Traditional Chinese Medicine

Objective:To investigate the effect of quercetin on ATP binding cassette transporter A1(ABCA1),liver X receptor(LXR),and proprotein convertase subtilisin/kexin type 9(PCSK9)expressions in apoE-knockout(ApoE-/-)mice.Methods:The high-fat diet-induced atherosclerosis(AS)in ApoE-/-mice was established.Thirtysix mice were divided into 3 groups using random number table method:model group(n=12),quercetin group(n=12),and atorvastatin group(n=12),with C57BL/6J mice of the same strain and age as the control group(n=12).Quercetin group and atorvastatin group were administrated with quercetin and atorvastatin by oral gavage,with doses of 12.5 and 4 mg/(kg・d),respectively.Animals in the control and model groups were given an equal volume of distilled water by oral gavage once per day for a total of 12 weeks.Western blot and immunohistochemical methods were employed to determine the aortic ABCA1,LXR-a and PCSK9 protein expressi on.En zyme linked imm uno sorbent assay method was used to detect the expressi on of serum total cholesterol(TC),triglyceride(TG),high density lipoprotein-cholesterol(HDL-C),low density lipoproteincholesterol(LDL-C),tumor necrosis factor-a(TNF-a),interleukin-6(IL-6),and IL-10,combined with tissue pathological exami nation.Results:ApoE-/-mice fed with a high-fat diet had no table atherosclerosis lesions,with reduced ABCA1,LXR-a and IL-10 levels(all P<0.01),elevated PCSK9,TNF-a and IL-6 expression,and increased TC and LDL-C con tents(all P<0.01).After querceti n in terventi on,the areas of AS plaques and the expressions of PCSK9,TNF-a and IL-6 were significantly reduced(all P<0.01),while the expressions of ABCA1 and LXR-a were increased significantly(all P<0.01).Conclusion:Quercetin effectively interfered with AS development by regulating the expressions of ABCA1,LXR-a and PCSK9 in ApoE,mice.

关键词： quercetin atherosclerosis ATP binding cassette transporter A1 liver X receptor proprotein convertase subtilisin/kexin type 9

In vitro Degradation and Biocompatibility of Ca-P Coated Magnesium Alloy

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Chemical Research in Chinese Universities 2013年第2期29卷 285-289页

作者： XIAO Xing zhu qing-san SU Ying-chao LI Guang-yu The First Hospital of Jilin University Changchun 130021 P. R. China College of Materials Science and Engineering Jilin University Changchun 130022 P. R. China

Calcium-phosphate compounds（Ca-P） coating was prepared on an Mg-A1 alloy（AZ60）. Biodegradation of Ca-P coated magnesium alloy was evaluated in simulated body fluid（SBF） by examining the changes in magnesium ion concentration and pH value, which indicated that the Ca-P coating on magnesium alloy strongly affected the cor- rosion of magnesium alloy. Osteoblast MC3T3-E1 cells were utilized to investigate the cellular eytocompatibility. The cytocompatibility was measured by carrying out a series of tests, such as cholecystokinin-octapeptide（CCK-8） test, alkaline phosphatase activity（ALP） test, cellular morphology of hematoxylin-eosin（HE） staining and the induc- tion of apoptosis. It was found that the cell function showed better in the Ca-P coated Mg-alloy extract than in the uncoated magnesium alloy extract. In summary, the results indicate that the Ca-P coating can improve the corrosion resistance of magnesium alloy and elevate cellular proliferation and differentiation of osteoblast MC3T3-E1 cells.

关键词： Ca-P coating Magnesium alloy In vitro

Implementing VLPR systems based on TMS320DM642

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Journal of Zhejiang University-Science A(Applied Physics & Engineering) 2007年第12期8卷 2005-2016页

作者： zhu Le-qing ZHANG san-yuan YE Xiu-zi School of Computer Science and Technology Zhejiang University Hangzhou 310027 China

This paper gives a practical schema for using DSP boards to construct Vehicle License Plate Recognition (VLPR) modules that could be embedded in any Intelligent Transportation System (ITS). Using DSP can avoid the heavy investment in dedicated VLPR system and improve the computational power compared to PC software environment. Low cost, high computational power, and high flexibility of DSP provide the License Plate Recognition System (LPRS) an excellent cost-effective solution to execute the major part of the recognition tasks. This paper describes a successful implementation of VLPR system based on Texas Instruments (TI)'s TMS320DM642. The DSP board acquires video (which could be output to a monitor for surveillance) from a camera, captures images from the video, locates and recognizes the license plates in images, and then sends the recognized results and related images after compression to a host PC through the network. Finally, the overall software is optimized according to the features of DM642 chip. Experiments showed that the DSP VLPR system performs well on the local license plates, and that the processing speed and accuracy can meet the requirement of practical applications.

关键词：许可证图象识别图象加工 DM642技术

Predictive risk factors for recollapse of cemented vertebrae after percutaneous vertebroplasty: A meta-analysis

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World Journal of Clinical Cases 2021年第12期9卷 2778-2790页

作者： Yi-Hang Ma Zhi-Sen Tian Hao-Chuan Liu Bo-Yin Zhang Yu-Hang zhu Chun-Yang Meng Xiang-Ji Liu Qing-san Zhu Department of Spine Surgery China-Japan Union Hospital of Jilin UniversityChangchun 130033Jilin ProvinceChina

BACKGROUND As one of the most common complications of osteoporosis,osteoporotic vertebral compression fracture(OVCF)increases the risk of disability and mortality in elderly patients.Percutaneous vertebroplasty(PVP)is considered to be an effective,safe,and minimally invasive treatment for OVCFs.The recollapse of cemented vertebrae is one of the serious complications of PVP.However,the risk factors associated with recollapse after PVP remain controversial.AIM To identify risk factors for the recollapse of cemented vertebrae after PVP in patients with OVCFs.METHODS A systematic search in EMBASE,MEDLINE,the Cochrane Library,and PubMed was conducted for relevant studies from inception until March 2020.Studies investigating risk factors for the recollapse of cemented vertebrae after PVP without additional trauma were selected for analysis.Odds ratios(ORs)or standardized mean differences with 95%confidence interval(CI)were calculated and heterogeneity was assessed by both the chi-squared test and the I-squared test.The methodological quality of the included studies was assessed according to the Newcastle-Ottawa Scale.RESULTS A total of nine case-control studies were included in our meta-analysis comprising 300 cases and 2674 controls.The significant risk factors for the recollapse of cemented vertebrae after PVP in OVCF patients were fractures located at the thoracolumbar junction(OR=2.09;95%CI:1.30 to 3.38;P=0.002),preoperative intravertebral cleft(OR=2.97;95%CI:1.93 to 4.57;P<0.00001),and solid lump distribution pattern of the cement (OR = 3.11;95%CI: 1.91 to 5.07;P < 0.00001).The analysis did not support that age, gender, lumbar bone mineral density,preoperative visual analogue scale score, injected cement volume, intradiscalcement leakage, or vertebral height restoration could increase the risk forcemented vertebra recollapse after PVP in OVCFs.CONCLUSIONThis meta-analysis suggests that thoracolumbar junction fractures, preoperativeintravertebral cleft, and solid lump cement distribution pattern are associatedwith

关键词： Vertebroplasty Osteoporotic vertebral compression fracture Risk factors Recollapse Cemented vertebrae Meta-analysis