1.Expression and significance of caspase-3 after spinal cord injury in rats:Selection of proper time window of intervention for secondary spinal cord injury
Chengla YI ; Anmin CHEN ; Weiguo XU ; Xiangjun BAI ; Xianzhou SONG
Chinese Journal of Tissue Engineering Research 2005;9(38):155-158
BACKGROUND: Caspase family is viewed as the executive factor of cell apoptosis. Neuronal apoptosis happens probably after spinal cord injury.OBJECTIVE: To observe the changes in caspase-3 expression after spinal cord injury in rats so as to probe into the relationship between it and neuronal apoptosis and provide the evidence on the prop e r time window of intervention on alleviating secondary spinal cord injury.DESIGN: Self-control and mutual-control were designed in animal experiment.SETTING: Department of Traumatic Surgery and Department of Orthopedics of Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology.MATERIALS: The experiment was performed in Experiment Room of Tongji Hospital affiliated to Tongji Medical College of Huazhong University of Science and Technology from January to December 2001, in which, 54SD rats were employed, of either sex, mass weighted varied from 220 to 250 g and provided from Animal Experimental Center of Tongji Medical College of Huazhong University of Science and Technology.rats were divided into the control and injury group. Laminectomy was only done on Ts and T9 in the control and the injury group was subdivided into 9 subgroups, in which, the materials were collected on the 4th and 8th hours and on the 1st, 2rd, 3rd, 7th 14th and 21st days successively, 6 rats in each one. After abdominal anesthesia with 30 g/L pentobarbitol sodium,sternal cord on T8 andT9 segments were exposed with Nystrom method and 50 g weight compressed the front middle region of the spinal cord of such segments with arch smooth metal pad 2.2 mn×5.0 mm for 5 minutes. After injury, artificial bladder urination was done 3 times at 10:00, 16:00and 22:00 successively everyday till the bladder reflex was established.cord was collected at various time spots after spinal cord injury. 4 pieces of spinal cord tissue masses from each group, about 8mm in length, were embedded with paraffin and sectioned continuously. Afterwards, HE staining, immunohistocheistry and TUNEL (TdT-mediated dUTP-biotin nick end labeling) were performed successively. Two rats were sacrificed on ice in each group and central tissue of injured spinal cord was placed in expression was assayed with immunohistochemistry method, neuronal apoptosis was assayed with TUNEL method and linear correlation was used to analyze the correlativity between caspase-3 expression and neuronal apoptosis.pase-3 expression after spinal cord injury in rats of each group.RESULTS: Six rats were maintained in each group and included in result optic microscope: Extensive hemorrhage appeared in 1 hour in injured segment. In 4 to 8 hours, spinal structure began destructive and a large amount of neuronal death appeared. In 24 hours, the destruction of spinal cord became severe and in 7 to 21days, the range of injury was defined with immunohistochemistry in rats of each group: Very few caspase-3 expressions (2.1±0.5) presented in neurons of spinal cord in normal rat. In 8hoursafter spinal cord injury, caspase-3 expression of positive neurons was increased remarkably (89.2±10.5) and up to the peak (189.6±12.7) in 3 days. Caspase-3 expression of positive cell and apoptotic cell appeared alexpression assayed with transcription-polymerase chain reaction (RT-PCR)in rats of each group: Caspese-3 mRNA (0.442±0.024) began increased in 4h, was up to the peak (0.634±0.028) in 48 hours and was restored to be normal (0.351±0.013) in 7 days, which appeared early than apoptosis, indicating positive correlation with the level of neuronal apoptosis (r=0.622).In the control and 4 hours group, stained cell was seen occasionally and positive cell appeared 8 hours later, mainly localized in gray matter. Afterwards, positive cell was increased and up to the peak in 3 days. In 7 days,positive cell of apoptosis and staining was decreased gradually in gray matter, mainly around the white matter. Little amount positive cells appeared on the 14th day and 21st day.CONCLUSION: In normal spinal cord tissue, caspase-3 existed in form of zymogen with very low activity. Caspase-3 is enhanced in expression after spinal cord injury in rats, expresses in large amount in 8 hours and is up to the peak in 24 to 48 hours, which is overlapped in time with positive apoptotic cell assayed with TUNEL and concerning to the localization, it is in conformity with positive apoptotic cell of spinal cord injury compared with positive cell of caspase-3. It is indicated that caspase-3 is involved in regulation of cell apoptosis after spinal cord injury. It is seen in this experiment that the time from spinal cord injury to the activation of caspase-3 is the time window of treatment for cell apoptosis intervened by spinal cord and alleviating secondary spinal cord injury. It is suggested that genetic intervention or specific caspase-3 inhibitor should be applied in 48 hours.
2.Relationship between the expression of MTA-1 gene and the metastasis and invasion in human osteosarcoma.
Chengla, YI ; Xinzhi, LI ; Weiguo, XU ; Anmin, CHEN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2005;25(4):445-7
To compare the expression level of metastasis associated-1 (MTA1) gene in high and low metastatic human osteosarcoma cell lines and examine the relationship of MTA1 expression and the metastasis potentiality of osteosarcoma cells, the expression of MTA1 in MG-63 osteosarcoma cell lines with high and low metastasis potential was detected by semiquantitative TR-PCR. Boyden chamber invasion assay was used to evaluate the invasive capacity in vitro in two osteosarcoma cell lines. The low metastasis MG-63 cells were transfected with MTA1 full-length cDNA expression plasmid by lipofectamine and the changes of MTA1 expression and in vitro invasion potential were examined after the transfection. Our results showed that MG63 cell line with high metastasis potential expressed significantly higher MTA1 than that of MG63 cells with low metastasis as reavealed by RT-PCR. The invasion potential of low metastasis MG63 cell line was increased after MTA1 gene transfection. It is concluded that there may be a relationship between MTA 1 and invasive potentiality of human osteosarcoma cells, and the mechanism of MTA1 in osteosarcoma metastasis and its possible role in associated gene therapy deserve further study.
Bone Neoplasms/*metabolism
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Bone Neoplasms/pathology
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Gene Expression Regulation, Neoplastic
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Histone Deacetylases/*biosynthesis
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Histone Deacetylases/genetics
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Neoplasm Invasiveness
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Neoplasm Metastasis
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Osteosarcoma/*metabolism
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Osteosarcoma/pathology
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RNA, Messenger/biosynthesis
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RNA, Messenger/genetics
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Repressor Proteins/*biosynthesis
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Repressor Proteins/genetics
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Tumor Cells, Cultured
3.Application of integrative treatment model and damage control theory in management of severe multiple injury
Fan YANG ; Xiangjun BAI ; Zhaohui TANG ; Chengla YI ; Zhanfei LI ; Wei GAO
Chinese Journal of Trauma 2009;25(9):843-846
Objective To investigate the clinical therapeutic effect of integrative treatment model and damage control theory in treatment of severe multiple injury. Methods A retrospective study was done on 116 patients with severe multiple injuries who were divided into integrative treatment plus damage control group, integrative treatment group and conventional treatment group. The relationship of correlative factors with mortality rate among three groups was compared before operation, during operation and after operation. Results All the indices in integrative treatment plus damage control group were better than other two groups before operation, during operation and after operation, with lower mortality rate (P<0.05). Conclusions Early use of integrative treatment model and damage control theory can obviously decrease complications, improve prognosis and decrease mortality rate for patients with severe multiple injury.
4.Early management of complication on severe multiple trauma
Fan YANG ; Xiangjun BAI ; Kaijun HU ; Zhaohui TANG ; Chengla YI ; Yiliu LIAO
Chinese Journal of Emergency Medicine 2009;18(6):628-631
Objective To analyze the efficacy of early management of eomplications in patients with multi-ple traumas by applying the damage control theory. Method All total of 116 patients admitted to Wuhan Tongji Hospital between Jan 2006 and Jan 2008 were included in this retrospective study and were classified according to treatment method. Overall, 55 cases underwent damage control operations and 61 cases underwent conventional management. The groups of patients were compared in terms of parameters such as length of operation time, time to resuscitate from shock, lethal triad of death (LTD), complications and treatment outcomes after operation. Results The length of operation time was (67.43±19.52) min, resuscitation time from shock was (6.77±3.16) h and LTD was (11.54±4.10) h in the damage control group, and (163.95±55.41) min, (22.51±11.65) h and (34.55±5.63) h, respectively, in the conventional group, with significant differences between the two groups (P< 0.01). The per capita complication and mortality rates were 227.27 % and 3.64 %, respectively, in the dam-age control group and 363.93 % and 9.84%, respectively, in the conventional group (P <0.05). Couclusions Here, we demonstrated that early management of patients with multiple traumas undergoing operations by apply-ing the damage control theory significantly decreased the operation time, and decreased time to resuscitation from shock and LTD. Furthermore, this approach significantly reduced the incidence of comphcations and the mortality rate of patients with severe multiple traumas.
5.Treatment of crush syndrome by emergency vacuum sealing drainage
Fan YANG ; Xiangjun BAI ; Chengla YI ; Zhanfei LI ; Chaohui YANG ; Bo LI
Chinese Journal of Trauma 2009;25(2):103-106
Objective To investigate the efficacy of emergency vacuum sealing drainage (VSD) in the treatment of crush syndrome. Methods A total of 102 patients with crush injury were randomly screened out and divided into emergency VSD group (n = 50), delayed VSD group (n = 22) and control group (n = 30). The variances of indices of crush syndrome after VSD between three groups were analyzed. Results The vital sign, urine volume, laboratory examination results were improved and the incidence rate of crush syndrome, complications and mortality rate were significantly decreased in emergency VSD group than those in delayed VSD group and control group (P < 0.05). For patients with crush syndrome, the incidence of complications, duration of hospital stay and mortality rate were significantly decreased in emergency VSD group and delayed VSD group than those in control group (P < 0.05). Conclusion Emergency VSD can suck the exudate of the necrotic tissues and the wounds, maintain draining wounds and prevent infection and hence is helpful to prevent and treat the crush syndrome.
6.Relationship between the Expression of MTA-1 Gene and the Metastasis and Invasion in Human Osteosarcoma
Chengla YI ; XinZhi LI ; Weiguo XU ; Anmin CHEN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2005;25(4):445-447
To compare the expression level of metastasis associated-1 (MTA1) gene in high and low metastatic human osteosarcoma cell lines and examine the relationship of MTA 1 expression and the metastasis potentiality of osteosarcoma cells, the expression of MTA1 in MG-63 osteosarcoma cell lines with high and low metastasis potential was detected by semiquantitative TR-PCR. Boyden chamber invasion assay was used to evaluate the invasive capacity in vitro in two osteosarcoma cell lines. The low metastasis MG-63 cells were transfected with MTA1 full-length cDNA expression plasmid by lipofectamine and the changes of MTA1 expression and in vitro invasion potential were examined after the transfection. Our results showed that MG63cell line with high metastasis potential expressed significantly higher MTA1 than that of MG63 cells with low metastasis as reavealed by RT-PCR. The invasion potential of low metastasis MG63 cell line was increased after MTA 1 gene transfection. It is concluded that there may be a relationship between MTA 1 and invasive potentiality of human osteosarcoma cells, and the mechanism of MTA1 in osteosarcoma metastasis and its possible role in associated gene therapy deserve further study.
7.Anterior expansion of sacral foramen and decompression of sacral plexus via lateral-rectus approach for sacral fractures complicated with sacral plexus injury
Qiubao ZHENG ; Shicai FAN ; Zhiyong HOU ; Chengla YI ; Qiguang MAI ; Tao LI ; Xiaorui ZHAN ; Fuming HUANG ; Xiangyuan WEN ; Yuancheng LIU
Chinese Journal of Orthopaedic Trauma 2021;23(1):47-54
Objective:To evaluate the anterior expansion of sacral foramen and decompression of sacral plexus via the lateral-rectus approach (LRA) in the surgical treatment of sacral fractures complicated with sacral plexus injury.Methods:From January 2013 to June 2018, 11 patients were treated at Department of Orthopaedics, The Third Hospital Affiliated to Southern Medical University for obsolete sacral fractures complicated with sacral plexus injury. They were 8 males and 3 females, aged from 17 to 54 years (average, 38 years). According to the Denis classification, all the sacral fractures belonged to Denis Zone Ⅱ. According to British Medical Research Council (BMRC) grading system, the nerve injury was complete damage in 2 cases and partial damage in 9. The mean time from injury to surgery was 6 months (range, from 0.7 to 12.0 months). After the sacroiliac joint was exposed via the LRA, the lumbosacral trunk was exposed and released between iliac vessels and the iliopsoas. Next, the S1 foramen was expanded and the S1 nerve root was released after separation of the median sacral artery and the internal iliac artery. Reduction and fixation of the sacroiliac joint was carried out for patients with unstable sacral fracture. X-ray and CT examinations of the pelvis were performed to evaluate fracture healing and neurological function recovery postoperatively.Results:Of this cohort of 11 cases, operation succeeded in 10 but failed in one whose sacral fracture was found to have completely healed with the S1 foramina totally occluded. The surgical time averaged 110 min (range, from 70 to 220 min) and the blood loss 1, 100 mL (range, from 450 to 2, 800 mL). Postoperative X-ray and CT examinations showed that the sacral foramens were expanded significantly without any complications. The follow-up time averaged 18 months (range, from 12 months to 4 years). By the BMRC grading system at the last follow-up, the neural function was completely recovered in 5 cases, partially recovered in 4 cases and not recovered in one.Conclusion:Significant anterior expansion of sacral foramen and decompression of sacral plexus via the LRA is a viable and effective alternative for treatment of sacral fractures complicated with sacral plexus injury.
8.Expression of vascular endothelial growth factor (VEGF) in human osteosarcoma cells transfected with adeno-associated virus-antisense VEGF.
Weiguo XU ; Anmin CHEN ; Yibei ZHANG ; Chengla YI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2004;24(3):279-283
The expression of protein vascular endothelial growth factor (VEGF) in osteosarcoma cells transfected with adeno-associated virus (rAAV)-antisense VEGF was studied to provide the foundation of osteosarcoma treatment through antivascularization. The rAAV-antisense VEGF at different doses (0, 20, 50, 100, 200, 240 microl) was transfected into osteosarcoma MG-63 cell. The cells and culture supernatants were collected before and after tansfection. The expression of VEGF protein was detected by using immunohistochemical staining (SP) and Western blot. SP and Western-blot tests revealed that the MG-63 Cells transfected with rAAV-antisense VEGF had less staining than those without transfection with rAAV-antisense VEGF, and the staining intensity was negatively correlated with the doses of genes. The corresponding A values of transfected genes with different doses of rAAV-antisense VEGF (0, 20, 50, 100, 200, 240 microl) were 86614 +/- 13776, 73245 +/- 15414, 61078 +/- 12124, 54657 +/- 10953, 39802 +/- 11308, 32014 +/- 15057 respectively, with the difference being significant (P<0.05). It was concluded that the expression of VEGF protein in MG-63 cells could be inhibited by rAAV-antisense VEGF.
Bone Neoplasms
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metabolism
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pathology
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Dependovirus
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genetics
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Gene Transfer Techniques
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Genetic Vectors
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Humans
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Oligodeoxyribonucleotides, Antisense
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pharmacology
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Osteosarcoma
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metabolism
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pathology
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Transfection
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Tumor Cells, Cultured
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Vascular Endothelial Growth Factor A
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antagonists & inhibitors
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biosynthesis
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genetics
9.Orthotopic osteochondral transplantation for the treatment of femoral head compression fracture: a case report and literature review
Meiqi GU ; Zhe XU ; Li HE ; Yangxing LUO ; Enzhi YIN ; Ruixiang CHENG ; Chengla YI
Chinese Journal of Orthopaedics 2023;43(2):131-135
Femoral head fracture is commonly seen in high-energy injury. However, compression fracture of femoral head is more rare. In most classifications of femoral head fracture, the compression is unusually involved. A case about acute traumatic dislocation of hip joint with compression fracture of femoral head is reported, involving a patient who hurt himself by riding electric bike and hitting the flower bed. He came to our hospital complaining of pain and limited motion of his right leg. The diagnosis of right hip anterior dislocation with the compression fracture of femoral head was confirmed by medical history, physical examination and imaging. Closed reduction of hip dislocation was performed in an emergency. Then we transplanted the bone cartilage from the non-weight-bearing area under the femoral head to the collapsed weight-bearing area, fixing it with countersunk hollow screws, and then the non-weight-bearing donor area was reconstructed with autogenous iliac bone, using surgical hip dislocation. The anatomical structure of the femoral head was therefore restored successfully during the operation. Three months after surgery, the X-ray showed that the femoral head was smooth and the cartilage graft was well fixed. Eight months after surgery, the patient gradually increased the bearing weight from partial to full according to his own condition, and there was no obvious pain in hip. After 24-month follow-up, we found the X-rays showed good reduction and fixation of the femoral head fracture. The CT scan showed no necrosis or cystic degeneration. He got well-active and passive movement in hip joint, and got no pain when walking with burden. For the patient with hip dislocation and compression fracture of femoral head, early joint reduction and non-weight-bearing osteocartilage transplantation can restore the anatomical structure of the weight-bearing area of the femoral head, to avoid traumatic osteoarthritis, and to improve the long-term quality of life of patients.
10.Pelvic unlocking closed reduction technique combined with HoloSight system in reducing and fixating Tile C1 pelvic fractures
Li HE ; Hua CHEN ; Chengla YI
Chinese Journal of Orthopaedics 2023;43(19):1308-1315
Objective:To explore the effectiveness of unlocking closed reduction technique (UCRT) combined with the "Transparent Orthopaedics" intelligent visualization system (HoloSight) in reducing and fixating Tile C1 type pelvic fractures.Methods:Data of 26 patients with Tile C1 pelvic fracture treated by trauma surgery in Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology from March 2019 to September 2022 were retrospectively analyzed. Among them, 13 patients were treated with the UCRT combined with the HoloSight system (HoloSight group), and 13 patients were treated with the UCRT (control group). There were 7 men and 6 women in the HoloSight group, with a mean age of 46.85±15.63 years. The pelvic fractures were classified as C1.1 in 2 cases, C1.2 in 2 cases, and C1.3 in 9 cases. The mean body mass index (BMI) was 24.08±4.15 kg/m 2. The preoperative vertical displacement was 1.80±0.76 cm and the posterior displacement was 0.80±0.63 cm. There were 9 men and 4 women in the control group, with a mean age of 38.38±13.82 years. The pelvic fractures were classified as C1.1 in 3 cases, C1.2 in 4 cases, and C1.3 in 6 cases. The mean BMI was 22.85±3.54 kg/m 2. The preoperative vertical displacement was 1.77±0.70 cm and the posterior displacement was 1.17±0.58 cm. The fracture reduction time, preoperative and postoperative fracture displacement, single screw fixation time, intraoperative reduction and fixation fluoroscopy times, visual analogue scale (VAS), Matta scoring, Majeed's score, and complications were recorded for each patient. Results:All patients were followed up for 13.65±5.06 months. The reduction time in the HoloSight group was 39.77±11.22 minutes, the single screw fixation time was 10.72±2.12 minutes, and the total fixation time was 37.15±12.12 minutes, which were significantly shorter than those in the control group, which were 67.46±16.67 minutes, 18.38±3.62 minutes, and 58.31±7.66 minutes, respectively ( t=4.97, 6.59, 5.32). The reduction and fixation fluoroscopy times were 4.38±1.33 times and 14.00±5.79 times in the HoloSight group, which were significantly less than those in the control group, which were 50.69±12.48 times and 47.77±19.34 times ( t=13.30, 6.03). The differences were statistically significant ( P<0.05). All patients' pelvic fractures healed with no significant difference ( t=0.47, P=0.644) in fracture healing time between the HoloSight group (4.92±1.66 months) and the control group (5.23±1.69 months). Postoperative Matta scoring showed that 13 patients in the HoloSight group were rated as excellent, 4 patients were rated as good, and the excellent and good rate was 100%. In the control group, 13 patients were rated as excellent and 4 patients were rated as good, with an excellent and good rate of 100%. There was no significant difference ( t=0.70, P=0.493) in preoperative VAS between the HoloSight group (6.08±1.32) and the control group (5.69±1.49). There was no significant difference ( t=1.09, P=0.286) in postoperative VAS between the HoloSight group (2.08±1.04) and the control group (2.62±1.45), and there was no significant difference in VAS improvement between the two groups ( t=1.15, P=0.262). There was no significant difference ( t=1.42, P=0.168) in Majeed's score at 6 months postoperatively between the HoloSight group (91.23±7.18) and the control group (86.38±9.98). Conclusion:HoloSight system combined with UCRT frame can effectively reduce the difficulty of fracture reduction and fixation, shorten the time of fracture reduction and fixation, and reduce the radiation to patients and doctors in the treatment of pelvic fractures.