1.Specific extraction and analysis of synthesized proteins from mesenchymal stem cells transplanted into the ischemic heart
Wan-Er LU ; Ying DAI ; Mu-Han TANG ; Kang WEI ; Shu-Jia CHEN ; Huai HUANG ; Jing LIN ; Hao-Rong PENG ; Li-Xuan ZHOU ; Dun-Zheng HAN
Medical Journal of Chinese People's Liberation Army 2025;50(8):991-998
Objective To specifically extract and analyze nascent proteins synthesized by bone marrow mesenchymal stem cells(BMSCs)after transplantation into ischemic hearts using a technique employing mutant methionyl-tRNA synthetase(MetRSL247G)for nascent protein labeling,in order to explore the potential mechanisms of action in BMSCs post-transplantation.Methods Point mutation at position 274 of the MetRS gene in BMSCs was induced via lentiviral infection to enable azidonorleucine(ANL)-mediated labeling of nascent proteins in BMSCs.The labeling efficiency was verified by means of fluorescent non-canonical amino-acid tagging(FUNCAT).Thirty healthy female C57BL/6J mice(8-10 weeks old)were divided into control and experimental groups,with 15 mice in each group.The acute myocardial infarction model was constructed by ligating the left anterior descending coronary artery in experimental group,while control mice underwent only thoracotomy without coronary ligation.After modeling,both groups received intramyocardial injections of MetRSL247G-modified BMSCs(MetRSL247G-BMSCs)at 3 different sites in the peri-infarct ischemic region.Mice were intraperitoneally injected with ANL every 6 hours for 4 times on postoperative days 0,2,and 6(n=5 for each time point)respectively,euthanized 24 h after the last injection,and cardiac tissues were isolated.The newly synthesized and labeled proteins produced by BMSCs after transplantation into the myocardium of experimental and control groups were collected,using an enrichment technique for ANL-tagged proteins and liquid chromatography-tandem mass spectrometry(LC-MS)analysis.Gene ontology(GO)analysis,Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis,protein-protein interaction(PPI)analysis,and heatmap visualization analysis were performed to identify differentially expressed proteins at the 3 time points and screen key pathways and genes.Results Under fluorescence microscopy,the MetRSL247G lentivirus-infected BMSCs were observed to be labelled with mCherry signals,confirming the successful construction of the MetRSL247G-BMSCs cell line.Green fluorescent signals were detected only in nascent proteins in culture medium containing both MetRSL247G-BMSCs and ANL,validating the sensitivity and specificity of the labeling method.GO analysis revealed that differentially expressed proteins were primarily involved in basic cellular biological processes such as extracellular exosome formation,extracellular matrix organization,and focal adhesion.KEGG and PPI analyses indicated that the differential proteins were mainly involved in complement and coagulation cascade pathway,actin cytoskeleton regulation pathway,and apoptosis pathway.Heatmap analysis showed significantly upregulated expression of anti-apoptosis and cell adhesion-related factors in experimental group on day 1(P<0.05),upregulated anti-apoptotic factors,pro-apoptotic factors,and cell adhesion-related factors on day 3(P<0.05),and upregulated anti-apoptotic factors,cell differentiation-related factors,and cell adhesion-related factors on day 7(P<0.05)compared with control group.Expression of apoptosis-inducing factor 1 was significantly downregulated on days 1 and 7(P<0.05).On day 3,most differentially expressed proteins,including anti-apoptosis factors(Protein S100-A11,Clusterin,Gelsolin),pro-apoptosis factor(Cathepsin B),cell differentiation-related factor(Transgelin-2),and cell adhesion-related factors(Cofilin-1,Periostin,Fibronectin)were significantly upregulated(P<0.05).Conclusions The MetRSL247G mutation enables BMSCs to incorporate ANL and synthesize labeled proteins,confirming the feasibility of this nascent protein labeling technique.Nascent proteins of BMSCs in ischemic myocardium primarily contribute to extracellular exosome secretion and extracellular matrix organization.BMSCs may adapt to and respond to ischemic and hypoxic environments by influencing complement and coagulation cascades,activating inflammatory factors,regulating actin cytoskeleton structure,and modulating apoptosis,thereby maintaining the survival of BMSCs.
2.Research of 3D printing model in analysis of anterior disc displacement with reduction
Bo-yu AN ; Fang-tong JIAO ; Shun ZHANG ; Wan-tong YU ; Ai-she DUN ; Zuo-qin ZHAO
Journal of Regional Anatomy and Operative Surgery 2025;34(8):653-657
Objective To deeply understand the anatomical basis of temporomandibular joint disorders,and explore and master the pathological characteristics of anterior displacement of temporomandibular joint disc through cadaveric dissection and 3D printing technology.Methods By dissecting the temporomandibular joints of 40 head and neck specimens,the bilateral structures of the temporomandibular joints were measured to obtain condyle and articular disc data.3D modeling was carried out using 3ds Max software and printed out a model of temporomandibular joint.The pathological model of the anterior disc displacement with reduction(ADDwR)was simulated by adjusting the opening degree,the position of the articular disk,and the position of the condyle of the model.Results The precise data of the right and left temporomandibular joint discs and condyle were successfully obtained by dissection and measurement.The thickness of the anterior band of left temporomandibular joint discs was(2.02±0.57)mm,the thickness of the middle band was(1.46±0.33)mm,the thickness of the posterior band was(3.00±0.46)mm,the anterior-posterior diameter was(9.60±0.72)mm,and the internal-external diameter was(17.73±0.84)mm.While the thickness of the anterior band of right temporomandibular joint discs was(1.84±0.35)mm,and the thickness of the middle band was(1.43±0.28)mm,the thickness of posterior band was(3.08±0.49)mm,the anterior-posterior diameter was(9.30±0.88)mm,and the internal-external diameter was(17.38±1.10)mm.The internal-external diameter of the left temporomandibular joint condyle was(18.97±0.41)mm,and the anterior-posterior diameter was(8.56±0.43)mm;the internal-external diameter of the right temporomandibular joint condyle was(18.86±0.75)mm,and the anterior-posterior diameter was(8.40±0.30)mm.The standard temporomandibular joint model was printed out by the measured data.The model was utilized to simulate the physiological state of temporomandibular joint under normal conditions,as well as the three pathological states of closed mouth,closed mouth to open mouth,and open mouth in the case of ADDwR.Conclusion The temporomandibular joint model can more intuitively present the changes of anatomical structure in reversible temporoman-dibular joint disorders,which is helpful for understanding and mastering the different classification of this disease.
3.Research of 3D printing model in analysis of anterior disc displacement with reduction
Bo-yu AN ; Fang-tong JIAO ; Shun ZHANG ; Wan-tong YU ; Ai-she DUN ; Zuo-qin ZHAO
Journal of Regional Anatomy and Operative Surgery 2025;34(8):653-657
Objective To deeply understand the anatomical basis of temporomandibular joint disorders,and explore and master the pathological characteristics of anterior displacement of temporomandibular joint disc through cadaveric dissection and 3D printing technology.Methods By dissecting the temporomandibular joints of 40 head and neck specimens,the bilateral structures of the temporomandibular joints were measured to obtain condyle and articular disc data.3D modeling was carried out using 3ds Max software and printed out a model of temporomandibular joint.The pathological model of the anterior disc displacement with reduction(ADDwR)was simulated by adjusting the opening degree,the position of the articular disk,and the position of the condyle of the model.Results The precise data of the right and left temporomandibular joint discs and condyle were successfully obtained by dissection and measurement.The thickness of the anterior band of left temporomandibular joint discs was(2.02±0.57)mm,the thickness of the middle band was(1.46±0.33)mm,the thickness of the posterior band was(3.00±0.46)mm,the anterior-posterior diameter was(9.60±0.72)mm,and the internal-external diameter was(17.73±0.84)mm.While the thickness of the anterior band of right temporomandibular joint discs was(1.84±0.35)mm,and the thickness of the middle band was(1.43±0.28)mm,the thickness of posterior band was(3.08±0.49)mm,the anterior-posterior diameter was(9.30±0.88)mm,and the internal-external diameter was(17.38±1.10)mm.The internal-external diameter of the left temporomandibular joint condyle was(18.97±0.41)mm,and the anterior-posterior diameter was(8.56±0.43)mm;the internal-external diameter of the right temporomandibular joint condyle was(18.86±0.75)mm,and the anterior-posterior diameter was(8.40±0.30)mm.The standard temporomandibular joint model was printed out by the measured data.The model was utilized to simulate the physiological state of temporomandibular joint under normal conditions,as well as the three pathological states of closed mouth,closed mouth to open mouth,and open mouth in the case of ADDwR.Conclusion The temporomandibular joint model can more intuitively present the changes of anatomical structure in reversible temporoman-dibular joint disorders,which is helpful for understanding and mastering the different classification of this disease.
4.Establishing a noninvasive canine model of lumbar intervertebral disc degeneration through cumulative axial load and compound motion
Tao GU ; Dike RUAN ; Dun WAN ; Xuangeng DENG ; Huagang SHI ; Simao SONG ; Wei HOU ; Wei CUI ; Yuliang SUN
Chinese Journal of Orthopaedics 2023;43(21):1450-1459
Objective:To investigate the feasibility of establishing a canine model of lumbar intervertebral disc degeneration through the application of cumulative axial load and a six-phase combined motion on the vertical sitting dog's lumbar spine.Methods:Twenty adult female grass dogs, each weighing 10.0±0.5 kg, were randomly divided into two groups, with 10 dogs in each group. In the model group, dogs were secured to an exercise machine in a vertical position, and six phases of lumbar spine movement (flexion and extension, left and right lateral flexion, left and right rotation, 45° each) were combined with a specific number of cycles under continuous axial load (245 N). In the control group, dogs were secured to the exercise machine in a vertical position without any intervention. Radiographic examinations were performed before and after 20,000, 50,000, 100,000, and 150,000 compound exercises in the model group. The disc height index (DHI) was measured through lateral X-ray, and MRI T2-mapping was used for quantitative analysis of intervertebral disc degeneration. When intervertebral disc degeneration was evident on MRI T2-weighted imaging (modified Pfirrmann system > Grade V), the combined motion was halted. Micro-CT quantitative analysis of bone mineral density (BMD) in the upper and lower endplates, trabecular bone structure, and histological staining (HE staining, "O" staining, Sirius red staining) were employed to verify and assess the degree of intervertebral disc degeneration.Results:After 50,000 compound exercises, mild degeneration of the intervertebral discs at L 6-7 and L 7S 1 was observed on T2-weighted imaging. With the accumulation of exercise load, the degree of degeneration progressively increased, reaching a moderate degree of degeneration after 100,000 composite exercises, and DHI began to decrease. Mild degeneration was also observed in the upper L 5-6 intervertebral disc. When the cumulative exercise volume reached 150,000 repetitions, the height of intervertebral spaces in the L 5-6, L 6-7, and L 7S 1 segments further decreased, and the intervertebral discs exhibited severe degeneration (improved Pfirrmann grading system Grades IV-VI). The upper L 4-5 intervertebral discs also displayed mild degeneration. Histological scores were as follows: L 5-6 (8.2±0.8), L 6-7 (9.5±0.7), and L 7S 1 (10.3±0.5), indicating a degree of degeneration in the order of L 5-6
5.Feasibility and clinical effect of gelatin sponge in reducing bone cement leakage in percutaneous kyphoplasty
Wei CUI ; Xiaoming XIONG ; Dun WAN ; Huagang SHI ; Tao GU ; Xuangeng DENG
Chinese Journal of Trauma 2020;36(10):899-904
Objective:To explore the feasibility and clinical efficacy of gelatin sponge packing in reducing bone cement leakage in percutaneous kyphoplasty (PKP).Methods:A retrospective case-control study was conducted in data of 171 patients (171 vertebrae) with monosegmental lumbar osteoporosis compressive fracture treated by PKP from January 2015 to December 2018 in Sichuan Orthopedic Hospital. There were 66 males and 105 females, with the age of (67.9±6.7)years (range, 60-87 years). There were 22 patients with T 10 fracture, 28 with T 11 fracture, 37 with T 12 fracture, 34 with L 1 fracture, 32 with L 2 fracture and 18 with L 3 fracture. A total of 80 patients were pre-filled with gelatin sponge before injection (Group A), and 91 patients were not filled with gelatin sponge before injection (Group B). The operation time, amount of bone cement, and rate of bone cement leakage were recorded. The change of anterior vertebral height, Cobb angle, visual simulation score (VAS) and Oswestry disability index (ODI) were compared before operation and at postoperative 1 day, 3 months, 6 months, 12 months. Results:All patients were followed up for 1-12 months [(12.8±0.6)months]. The operation time in Group A and B was (48.3±1.2)minutes and (42.3±1.3)minutes ( P<0.05). The amount of bone cement in Group A and B was (5.4±0.8)ml and (5.6±0.7)ml ( P>0.05). The incidence of bone cement leakage in Group A and B was 11% (9/80) and 26% (24/91) ( P<0.05). There was no significant difference between the two groups in the anterior height of injured vertebrae, change of Cobb angle, VAS and ODI before and after operation ( P>0.05). Conclusion:Gelatin sponge can reduce the rate of bone cement leakage in PKP for the treatment of thoracolumbar osteoporosis compressive fracture, and has similar effect with PKP in correcting kyphosis, alleviating pain and improving life quality.
6.Preliminary application of CPC/PMMA composite bone cement in kyphoplasty for the elderly.
Xuan-Geng DENG ; Xiao-Ming XIONG ; Wei CUI ; Tao GU ; Dun WAN ; Hua-Gang SHI ; Xing CHEN ; Si-Mao SONG ; Wei HOU ; Guo-Long MEI ; Wen-Bing JIANG
China Journal of Orthopaedics and Traumatology 2020;33(9):831-836
OBJECTIVE:
From the perspective of clinical application to analyze the effectiveness and reliability of CPC/PMMA bone cement in percutaneous kyphoplasty (PKP) for the treatment of elderly patients with osteoporotic thoracolumbar fractures.
METHODS:
A retrospective analysis was performed on 62 patients with osteoporotic compression fracture of single-vertebral thoracic or lumbar segment who underwent PKP surgery and had a bone density less than or equal to -3.0 SD from February 2016 to December 2016. Among them, 23 patients were in CPC/PMMA group, with an average age of (77.6±2.2) years old, 39 patients in PMMA group, with an average age of (77.1±1.1) years old. The indexes between two groups were compared, including the visual analogue scale (VAS), height ratio of anterior vertebra (AVHR), local Cobb angle, cement leakage, new adjacent vertebral fracture(NAVF).
RESULTS:
There were no significant difference in gender, age, follow-up time and preoperative VAS, AVHR, local Cobb angle between two groups (>0.05), at the 1 day after operation, VAS, AVHR, local Cobb angle in all patients got obvious improvement (<0.05), which was no significant difference at 1 day after operation and final follow-up (>0.05). At the same time, there was no statistically significant difference in the incidence of new adjacent vertebral fracture and cement leakage (>0.05). The pain in both groups continued to improve at follow up after operation (<0.05), the local Cobb angle increased (<0.05) and AVHR decreased slightly (<0.05). However, the images of conventional methods (X-ray or CT) could not find signs about CPC degeneration and new bone ingrowth.
CONCLUSION
CPC/PMMA composite bone cement is safe and reliablein PKP for treatment of elderly patients with osteoporotic thoracolumbar fractures, which can effectively relieve pain and maintain vertebral body stability. It has the same curative effect as PMMA bone cement. It was worthy to research more in future, although no direct evidences support the CPC/PMMA composite bone cement can reduce the incidence of adjacent vertebral fracture, CPC degeneration or new bone ingrowth.
Aged
;
Bone Cements
;
Dinucleoside Phosphates
;
Fractures, Compression
;
Humans
;
Kyphoplasty
;
Osteoporotic Fractures
;
Polymethyl Methacrylate
;
Reproducibility of Results
;
Retrospective Studies
;
Spinal Fractures
;
Treatment Outcome
;
Vertebroplasty
7.Application of enhanced recovery after surgery in postoperative rehabilitation of osteoporotic lumbar compression fractures with percutaneous vertebroplasty or percutaneous kyphoplasty.
Xing CHEN ; Dun WAN ; Xiao-Ming XIONG ; Hua-Gang SHI ; Xuan-Geng DENG ; Tao GU ; Si-Mao SONG ; Wei HOU ; Qing-Long LI
China Journal of Orthopaedics and Traumatology 2020;33(12):1179-1183
OBJECTIVE:
To study effects of postoperative regular training of core muscle strength guided by the concept of enhanced recovery after surgery (ERAS) on the rehabilitation of elderly patients with osteoporotic lumbar vertebral compression fracture after vertebroplasty (PVP) and kyphoplasty(PKP).
METHODS:
Ninety-four elderly patients with osteoporotic lumbar compression fractures who underwent PKP or PVP from January 2016 to January 2018 and met inclusion criteria were divided into observation group and control group. All the patients were treated with routine anti osteoporosis therapy after operation. There were 47 patients in the observationgroup, including 18 males and 29 females, with an average age of (62.62±3.21) years old;in the control group, there were 47 cases, including 17 males and 30 females, with an average age of (62.38±2.84) years old. The patients in the control group were trained by traditional way, and the patients in observation group were instructed to conduct regular training of core muscle strength according to ERAS concept. The patients were followed up for 1, 3 and 6 months after operation. Patients' conditions were quantitatively evaluated according to Barthel scale, JOA low back pain score and Oswestry Disability Index, and the differences in treatment effects between two groups were statistically analyzed and compared.
RESULTS:
All the patients were followed up, and the Barthel scale, JOA low back pain score and Oswestry Disability Index score of the observation group were all better than those of the control group on the 1st and the 3rd months after surgery(
CONCLUSION
Early regular core strength training has a positive effect on early functional recovery and improvement of life ability after PKP or PVP for elderly patients with osteoporotic lumbar compression fractures, which is in line with the concept of accelerated rehabilitation surgery.
Aged
;
Enhanced Recovery After Surgery
;
Female
;
Fractures, Compression/surgery*
;
Humans
;
Kyphoplasty
;
Male
;
Middle Aged
;
Osteoporotic Fractures/surgery*
;
Spinal Fractures/surgery*
;
Treatment Outcome
;
Vertebroplasty
8.Application of Mobile Assessment in Aquatic Rehabilitation Assessment for Patients with Spinal Cord Injury
Yao CUI ; Fang CONG ; Jian-jun LI ; Ming ZENG ; Dong-yang LI ; Long JIN ; Feng-shan SI ; Bin YAO ; Wei JIA ; Dun-wu XIAO ; Kai ZHANG
Chinese Journal of Rehabilitation Theory and Practice 2019;25(3):341-346
Objective:To explore the application of mobile medical technology in aquatic rehabilitation assessment for patients with spinal cord injury. Methods:From November, 2017 to April, 2018, 72 patients with spinal cord injury accepted aquatic exercise were randomly divided into control group (
9.Clinical comparative study of short-segment and long-segment fixation for single-segment thoracic and lumbar spine III stage Kümmell disease.
Xuan-Geng DENG ; Xiao-Ming XIONG ; Dun WAN ; Hua-Gang SHI ; Wei CUI ; Xing CHEN ; Guo-Long MEI ; Si-Mao SONG ; Wei HOU
China Journal of Orthopaedics and Traumatology 2019;32(7):598-603
OBJECTIVE:
By comparing the clinical efficacy of short-segment and long-segment fixation for single-segment thoracic and lumbar spine III stage Kümmell disease to explore a more suitable fixed segment for the disease.
METHODS:
The clinical data of 46 patients with single-segment thoracic and lumbar spine III stage Kümmell disease treated from July 2013 to December 2016 were retrospectively analyzed. Forty-six patients were divided into short-segment fixation group(one vertebra above and below the diseased vertebra) and long-segment fixation group(two vertebrae on the upper and lower of the diseased vertebra) according to different methods of cement stick fixation. There were 25 patients in the short-segment fixation group, including 9 males and 16 females, with an average age of (75.3±4.5) years old, lumbar spine bone mineral density T-value of (-3.1±0.3) g/cm³, follow-up time of (13.0±2.3) months; there were 21 patients in long-segment fixation group, 6 males and 15 females, with an average age of (74.5±3.9) years old, lumbar spine bone mineral density T-value of (-3.2±0.3) g/cm³, follow-up time of (14.7±3.6) months.The gender, age, follow-up time, operation time, intraoperative blood loss, cement leakage, and the rate of adjacent vertebrae fractures were compared between two groups, as well as pain VAS score, ODI, and kyphosis angle before and after surgery.
RESULTS:
There were no significant differences in age, gender, bone density, pain VAS score, ODI, and kyphosis between two groups before surgery. The operation time and intraoperative blood loss of short-segment fixation group were less than that of long-segment fixation group. The pain VAS score, ODI and kyphosis of the two groups were significantly improved at 7 days after the operation and at the latest follow-up, there was no significant difference between two groups. There were no significant differences in bone cement leakage(9/25 vs 11/21) and adjacent vertebrae fractures(4/25 vs 3/21).
CONCLUSIONS
Both long-segment fixation and short-segment fixation can effectively relieve pain, correct kyphosis, improve functional index, and achieve better clinical results, but short-segment fixation has less operation time and less intraoperative blood. So single-segment thoracic and lumbar spine III stage Kümmell disease does not need to extend the fixed segment, short-segment fixation is more in line with clinical needs and worthy of further study.
Aged
;
Female
;
Fracture Fixation, Internal
;
Humans
;
Kyphosis
;
Lumbar Vertebrae
;
Male
;
Retrospective Studies
;
Spinal Fractures
;
Thoracic Vertebrae
;
Treatment Outcome
10.Observation of Proliferation of Human Umbilical Vein Endothelial Cells Treated with Serum Containing Yang-warming, Blood-activating, and Toxin-removing Recipe Sampled Under Different Conditions
Rui-Li HUANG ; Zhi-Wen LI ; Shuang CHEN ; Chun-Xiao ZHAO ; Wan-Hong LIANG ; Yong-Dun HONG
Journal of Guangzhou University of Traditional Chinese Medicine 2018;35(2):284-289
Objective To study the influences of Yang-warming, Blood-activating, and Toxin-removing Recipe(YBTR)-containing serum sampled under different conditions on the proliferation of human umbilical vein endothelial cells(HUVECs). Methods HUVECs were cultured in vitro for the experiment. Pharmacological serum for the experiment was prepared as follows: Thirty-two SD rats (male in half) were randomly divided into 4 groups, namely normal saline group and low-, middle-, and high-dose YBTR groups (at the intragastric dosage of 10.35, 31.05, 93.15 g·kg-1 respectively, twice a day). The pharmacological serum was taken from one female rat and one male rat in various groups at 4 time points, i.e. at hour 1, 2 after first intragastric administration on the fourth feeding day, and at hour 1, 2 after first intragastric administration on the sixth feeding day(abbreviated as D3H1, D3H2, D5H1, D5H2 respectively). The effects of YBTR-containing serum on the proliferation of HUVECs were observed by CCK-8 assay method. Results The difference of proliferation-inhibition rate of HUVECs was statistically significant after treated with YBTR-containing serum prepared from rats of different genders at different time(Pgender=0.000<0.01, Ptime=0.000<0.01). The difference of interaction of time and gender was also significant (Ptime × gender=0.001<0.01), and the effect at D3H1 and D5H1 varied with the gender (PD3H2×gender = 0.000 < 0.01, PD5H1×gender = 0.002 < 0.01). The inhibitory action of YBTR-containing serum became stronger with the increase of the dosage of serum of female rat at D3H2, and the inter-group difference was statistically significant (P < 0.05), the effect showing concentration-dependent tendency. The inhibition of HUVECs proliferation reached the peak after treated by various doses of YBTR-containing serum from the female rat at D3H2, while the inhibition arrived to the peak after treated by low- and middle-dose YBTR-containing serum from the male rat at D5H1, and the inhibition arrived to the peak in the group of high-dose YBTR-containing serum from the male rat at D3H1. Conclusion The inhibitory action of YBTR-containing serum on the proliferation of HUVECs was stronger when the serum was taken from the female rat at D3H2.

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