Mechanism of epidural scar hyperplasia after laminectomy induced by blood siltation / 中华骨科杂志

Objective:To explore the mechanism of epidural scar tissue hyperplasia induced by erythrocyte rupture and release of interleukin-33 (IL-33) after laminectomy in mice.Methods:In the zoological experiment, the operation group (Laminectomy) and the sham operation group were set, and HE staining and Masson staining were performed to test for blood accumulation in the operation area after laminectomy in mice. Then 12 wild-type mice with 6-8 week old were selected and divided into 4 groups: the sham operation group, the operation group (normal saline control), the pure red blood cell intervention operation group, the whole blood intervention operation group. The normal saline (100 mg/kg) was injected into the postoperative area. The red blood cells or whole blood with the same volume were injected into the postoperative area in the pure red blood cell intervention group and the whole blood intervention group. The postoperative recovery of mice in each group was observed. The levels of fibronectin in the postoperative scar tissues of mice in four groups were detected by western blot technology, and the degree of postoperative epidural scar hyperplasia was directly observed by immunohistochemistry. In the cytological experiment, the wild-type mouse erythrocyte normal saline group, the control group of IL-33 knockout mouse erythrocyte normal saline, the wild-type mouse erythrocyte lysis group, and the IL-33 knockout mouse erythrocyte lysis group were set. The levels of IL-33 in the red blood cells of four groups were detected by western blot. Then, a blank wild-type mouse erythrocyte control group, a wild-type mouse relative to the control group (only secondary antibody added to test for non-specific binding), a wild-type mouse erythrocyte group and an IL-33 knockout mouse erythrocyte group (to test for antigen specificity of the primary antibody) were set. Immunofluorescence staining was performed on the erythrocytes of four groups and the level of IL-33 was detected by flow cytometry.Results:HE staining and Masson staining after laminectomy showed that there was blood stasis in the local incision area of mice in the operation group. The epidural scar hyperplasia in the incision area of mice after whole blood or red blood cells intervention was higher, especially in the whole blood intervention group. IL-33 expression was almost undetectable in the wild-type erythrocyte normal saline control group, the IL-33-knockout erythrocyte normal saline control group, and the IL-33-knockout erythrocyte lysis group, while significant IL-33 expression was detectable in the wild-type erythrocyte lysis group. Immunofluorescence staining showed that IL-33 was expressed in and on the erythrocyte membrane of wild-type mice, while non-specific expression of IL-33 or a very small amount of IL-33 was almost undetectable in the other three groups. The immunofluorescence intensities of IL-33 in the four groups were 0.62±0.41, 60.17±4.39, 16.78±7.43 and 0.61±0.03, respectively ( F=281.90, P<0.001). The expression of IL-33 in the erythrocyte group of wild-type mice was the highest ( P<0.05). According to the results of flow cytometry, except for the trace amount of IL-33 detected in the wild-type mouse erythrocyte group, the expression of IL-33 in the other three groups was basically 0. The ratios of fibronectin to β-actin in the modeling area of the four groups gradually increased, and the ratios were 0.79±0.09, 1.26±0.23, 1.79±0.05 and 2.29±0.58, respectively, and the differences were statistically significant ( F=12.86, P=0.002). Fibronectin in the operation area of the three operation groups (normal saline control group, red blood cell intervention group and whole blood intervention group) was significantly higher than that of the sham operation group. The immunohistochemical staining results of fibronectin in the modeling area of the four groups were the same as those in western blot experiment. The average optical density values of fibronectin in each group were 0.09±0.01, 0.18±0.01, 0.22±0.01 and 0.24±0.01, respectively, and the difference was statistically significant ( F= 210.7, P<0.001). Conclusion:There is indeed blood accumulation in the surgical area after laminectomy in mice, and it can aggravate the hyperplasia of epidural scar tissue. Erythrocyte is the main component in blood, and there is a large amount of IL-33 expression in the inner and outer membrane of erythrocyte membrane. The mechanism of promoting the proliferation of epidural scar tissue may be related to the release of IL-33 by erythrocyte lysis.

Predictive Factors for Adjacent Vertebral Fractures After Percutaneous Kyphoplasty in Patients With Osteoporotic Vertebral Compression Fracture.

BACKGROUND: Adjacent vertebral fracture (AVF) seemed to be a frequent and severe complication in osteoporotic vertebral compression fracture (OVCF) patients receiving percutaneous vertebroplasty  or percutaneous kyphoplasty (PKP), resulting in poor long-term outcome and recurrence of pain-related symptoms. Nonetheless, its mechanism remains unclear. OBJECTIVES: To investigate the potential predictor of AVF after PKP and figure out whether the intervertebral disc plays a role during the process of AVF. STUDY DESIGN: Retrospective study. SETTING: Department of Orthopedic, an affiliated hospital of a medical university. METHODS: Clinical data of OVCF patients receiving PKP were reviewed in our hospital from January 2016 to December 2020. Four hundred and forty-five patients were recruited who met the abovementioned criteria in this study. The clinical data, including age, gender, bone mineral density (BMD), vertebral height, vertebral kyphosis angle, cement volume, cement distribution, as well as adjacent disc degeneration extent, were recorded for each patient. Independent-sample t tests and chi-squared tests were performed to compare these indexes. Bivariate correlation tests and multiple linear regression analyses were performed among potential predictors. Receiver operator characteristic (ROC) analysis and Kaplan-Meier plotter were applied to evaluate the diagnostic efficiency of parameters for predicting the occurrence of AVF. RESULTS: Patients in both groups gained obvious improvements in symptomatic and radiographic indexes after first PKP. Statistically significant difference (P < 0.05) was only found between 2 groups with respect to BMD, kyphosis angle at last follow-up before second PKP, cement distribution, and disc degeneration grade. The ROC analysis showed that BMD = 15.5° was highly predictive of AVF after PKP (sensitivity, 92.2%; specificity, 24.6%; area under curve, 0.569, P = 0.109). Statistically significant difference of AVF incidence amongst patients with different cement distribution (P = 0.018) and similar trend was also found amongst patients with different disc degeneration (P = 0.000). Statistically significant difference was noted in terms of disc degeneration grade between 2 adjacent discs in AVF group. LIMITATIONS: The main limitation is the retrospective nature of this study. CONCLUSIONS: The risk of AVF should be focused, especially when OVCF patients with the following predictors: (1) BMD < -3.45; (2) kyphosis angle at last follow-up > 15.5°; (3) I or II cement distribution; and (4) IV or V disc degeneration. More prophylactic treatment should be prescribed for these patients to avoid the occurrence of AVF.

IL-33 Promotes ST2-Dependent Fibroblast Maturation via P38 and TGF-β in a Mouse Model of Epidural Fibrosis

BACKGROUND@#Recent evidence suggests that IL-33, a novel member of the IL-1b family, is involved in organ fibrosis. However, the roles of IL-33 and its receptor ST2 in epidural fibrosis post spine operation remain elusive. @*METHODS@#A mouse model of epidural fibrosis was established after laminectomy. IL-33 in the wound tissues post laminectomy was measured with Western blotting, ELISA and imaging. The fibroblast cell line NIH-3T3 and primary fibroblasts were treated with IL-33 and the mechanisms of maturation of fibroblasts into myofibroblasts were analyzed. To explore roles of IL-33 and its receptor ST2 In vivo, IL-33 knockout (KO) and ST2 KO mice were employed to construct the model of laminectomy. The epidural fibrosis was evaluated using H&E and Masson staining, western-blotting, ELISA and immunohistochemistry. @*RESULTS@#As demonstrated in western blotting and ELISA, IL-33 was increased in epidural wound tissues post laminectomy. The immunoflurosence imaging revealed that endothelial cells (CD31 + ) and fibroblasts (a-SAM +) were major producers of IL-33 in the epidural wound tissues. In vitro, IL-33 promoted fibroblast maturation, which was blocked by ST2 neutralization antibody, suggesting that IL-33-promoted-fibroblasts maturation was ST2 dependent. Further, IL-33/ ST2 activated MAPK p38 and TGF-β pathways. Either p38 inhibitor or TGF-β inhibitor decreased fibronectin and a-SAM production from IL-33-treated fibroblasts, suggesting that p38 and TGF-β were involved with IL-33/ST2 signal pathways in the fibroblasts maturation. In vivo, IL-33 KO or ST2 KO decreased fibronectin, a-SMA and collagen deposition in the wound tissues of mice that underwent spine surgery. In addition, TGF-β 1 was decreased in IL-33 KO or ST2 KO epidural wound tissues. @*CONCLUSION@#In summary, IL-33/ST2 promoted fibroblast differentiation into myofibroblasts via MAPK p38 and TGF-β in a mouse model of epidural fibrosis after laminectomy.

Is unilateral transverse process-pedicle percutaneous kyphoplasty a better choice for osteoporotic thoracolumbar fractures in the old patients?

BACKGROUND: A few reports have shown that unilateral transverse process-pedicle percutaneous kyphoplasty is a good choice for patients with osteoporotic vertebral compression fracture (OVCF). However, this issue remains controversial and the related comprehensive research was lacked. METHODS: A retrospective study was conducted on patients receiving PKP surgery for OVCF. Patients were divided into three groups according to surgical approach. Symptom and radiographical evaluation were performed preoperatively, 1-month postoperatively, 1-year postoperatively and follow-ups. And follow-ups were repeated every year. Visual Analogue Scale Score (VAS), Oswestry Disability Index (ODI) scores, anterior vertebral height, coronal Cobb angle and sagittal Cobb angle was determined and compared among three groups. RESULTS: Totally 447 patients were included with an average age of 76.6 ± 7.2 years old. UTP showed significantly shorter surgical duration (p < 0.001), lower cement volume (p < 0.001) but higher cement leakage proportion (p = 0.044). No significant statistical difference was found in terms of improvement rates among three groups. Besides, it was notable that the a significantly higher coronal Cobb angle was observed in UTP group, and a about 4°coronal correction was found after UTP PKP. CONCLUSION: UTTP PKP could achieve similar symptoms relief and kyphosis correction as UTP and BTP PKP. However, it had shorter surgical time and less radio exposure than BTP PKP, lower risk of cement leakage and higher proportion of bilaterally cement distribution than UTP PKP. It seemed to be a better choice for patients with OVCF. In addition, we found that UTP PKP was especially fit for OVCF patients with asymmetrical vertebral compression.

3D printed guide template used in osteotomy for malunion of tibial fracture / 中华创伤骨科杂志

Objective:To evaluate the personalized 3D printed guide template used in the osteotomy for malunion of tibial fracture.Methods:A retrospective analysis was conducted of the 30 patients who had been treated for malunion of tibial fracture at Department of Orthopaedics, The First Affiliated Hospital to Zhengzhou University from January 2010 to January 2018. Of them, 15 used a personalized 3D printed guide template in the osteotomy (3D printing group). They were 9 males and 6 females, with an age of 46.3 year±8.2 years. The fracture malunion was located in the upper and middle tibia in 11 cases, in the lower tibia in 4 cases, on the left side in 6 cases and on the right side in 9 ones. There were 8 cases of varus deformity and 7 ones of valgus deformity. Their preoperative fracture deformity angle was 24.3°±5.5°. The other 15 patients were treated with conventional surgery (conventional group). They were 10 males and 5 females, with an age of 47.1 years±6.0 years. The fracture was located in the upper and middle tibia in 12 cases, in the lower tibia in 3 cases, on the left side in 5 cases and on right side in 10 cases. There were 7 cases of varus deformity and 8 ones of valgus deformity. Their preoperative fracture deformity angle was 22.5°±5.4°. The 2 groups were compared in terms of preoperative baseline data, operation time, intraoperative blood loss and postoperative recovery of the alignment of lower limb.Results:There were no significant differences in the preoperative baseline data between the 2 groups, showing comparability ( P>0.05). The 3D printing group was followed up for an average of 12 months while the conventional group for an average of 10 months. The operation time for the 3D printing group was significantly shorter than that for the conventional group(102.2 min±13.0 min versus 137.9 min ±10.5 min), the intraoperative blood loss for the former significantly less than that for the latter (77.3 mL ± 39.7 mL versus 163.3 mL ± 35.2 mL), and the postoperative malunion angle in the former significantly smaller than that in the latter (1.9°±0.4° versus 3.2°±0.9°) (all P< 0.05). The last follow-ups revealed no implant failure or re-malunion but fine healing of the osteotomy sites and good recovery of the alignment of lower limb in the 2 groups. Conclusion:A personalized 3D printed guide template used in the osteotomy for malunion of tibial fracture is an effective aid because it can facilitate precise osteotomy, reduce operation time and intraoperative blood loss and help correct the alignment of lower limb, leading to good short-term surgical outcomes.

Effect of lncRNA loc730101 on the biological characteristics of osteosarcoma U2OS cells through Wnt/β-catenin signal pathway / 中华显微外科杂志

Objective:To investigate the effect of long-chain non coding RNA (lncrna) loc730101 in the proliferation, invasion and migration of U2OS cells, and its mechanism.Methods:From February, 2019 to October, 2019, U2OS cells cultured in vitro were divided into control group (normal culture), negative group (transfection nega- tive control), and interference group (transfection of interference sequences targeting LOC730101). The expression of LOC730101 in cells was detected by reverse transcription-polymerase chain reaction (RT-PCR). Cell proliferation ac tivity was tested by methylthiazolyldiphenyl-tetrazolium bromide (MTT) method. Cell clone formation rate was mearused by plate clone formation test. Cell cycle distribution was tested by flow cytometry. Cell invasion and migra- tion were examed by Transwell chamber. The expression levels of epithelial-mesenchymal transition-related proteins Vimentin, N-cadherin, E-cadherin, and Wnt/β-catenin signaling pathway related proteins in cells β-catenin, c-Myc, cyclin D1(CyclinD1) and matrix metalloproteinase-7(MMP-7) proteins were detected by Western blotting method. The date was statistical analysed and considered as statistically significant when P<0.05. Results:Compared with the control group, the expression level of LOC730101 (0.25±0.03 and 1.00±0.06) in interference group and control group, respectively. The same below), cell survival rate [(57.65±3.26)% and (100.00±7.39)%], clone formation rate [(13.03± 2.12)% and (25.35±3.58)%], number of invasive cells(51.36±3.48 and 92.85±6.62), number of migrating cells (77.15± 5.05 and 136.92±15.35), the percentage of cells in S phase [(20.54±2.15)% and (28.15±2.38)%] and G 2/M phase [(16.87±2.12)% and (23.36±3.12)%], as well as the expression level of Vimentin (0.52±0.04 and 1.17±0.13), N-cadherin (0.31±0.03 and 0.65±0.04), β-catenin (0.42±0.03 and 0.73±0.04), c-Myc (0.29±0.03 and 0.65±0.03), CyclinD1 (0.26± 0.02 and 0.58±0.04), MMP-7 protein (0.55±0.03 and 0.86±0.06) was decreased significantly ( P<0.05), while the per- centage of cells in G 0/G 1 phase [(62.62±5.15)% and (48.46±3.65)%] and the expression level of E-cadherin protein(0.82± 0.06 and 0.38±0.03) were increased significantly in the interference group ( P<0.05). But there was no significant differ- ence in each index in the negative group ( P>0.05). Conclusion:Reduce the regulation of LOC730101 can inhibit the proliferation, invasion and migration of U2OS cells, and its mechanism may be related to the inhibition of Wnt/β-catenin signaling pathway.

Research of the expression and significance of syntaxin 8 in non-small cell lung cancer tissues / 天津医药

Objective To investigate the expression and significance of syntaxin 8 (STX8) in non-small cell lung cancer (NSCLC),and analyze the relationship between STX8 expression and clinicopathological features of NSCLC.Methods Seventy samples of NSCLC and 70 samples of pericancerous tissues were collected for immunohistochemistry staining,and another 10 samples of NSCLC and pericancerous tissues were used for RNA and protein extraction,qRT-PCR was performed to detect the expression of STX8 mRNA,and Western blot assay was adopted to detect the expression of STX8 protein.The relationship between STX8 expression and clinicopathological features of NSCLC was analyzed.Results Results of qRT-PCR showed that the expression of STX8 mRNA was significantly higher in NSCLC tissues than that in the pericancerous tissues (3.962 5±0.487 3 vs.0.538 2±0.097 5,t=21.797,P＜0.01).Immunohistochemistry results showed that the high expression rate of STX8 was significantly higher in NSCLC tissues than that in the pericancerous tissues [71.43％ (50/70) vs.38.57％ (27/70),x2=15.267,P＜0.01].Western blot results showed that the expression of STX8 protein was significantly higher in NSCLC tissues than that in the pericancerous tissues (2.496 1±0.362 5 vs.0.340 2±0.119 1,t=17.876,P＜0.01).The high expression rate of STX8 was significantly different in different histological types of NSCLC (P ＜0.05).The high expression rate of STX8 was higher in squamous cell carcinoma and adenocarcinoma than that in adenosquamous carcinoma,and the high expression rate of STX8 was not observed in large cell carcinoma.There were no significant differences in expressions of STX8 in NSCLC patients with different gender,ages,tumor diameters,TNM stages and with or without lymph node metastasis (P＞0.05).Conclusion A high expression of STX8 in NSCLC tissues may be correlated with the occurrence and development of NSCLC.

Analgesia after arthroscopic surgery:Randomly controlled Meta-analysis on intra-articular injection of bupivacaine placebo / 中国组织工程研究

BACKGROUND:Pain after arthroscopic treatment can not only affect the patient’s life quality, and is not conducive to the early reasonable exercise and functional recovery of the patients after treatment. Up to 2012, there are 18 randomized placebo-control ed trials on intra-articular injection of bupivacaine for analgesia after arthroscopic surgery, but the results are different. OBJECTIVE:To examine the efficacy and safety of intra-articular injection of bupivacaine in the management of pain after arthroscopic surgery through randomized placebo-control ed trials. METHODS:The MEDLINE database, Cochrane Central Register and Google Scholar database were retrieved for the randomized control ed trials on intra-articular injection of bupivacaine in the management of pain after arthroscopic surgery up to April 2012. The key words were“bupivacaine, intra-articular, arthroscopic, postoperative pain, placebo”. RESULTS AND CONCLUSION:Eighteen studies (n=934) were included (461 cases in bupivacaine group and 473 cases in the placebo control group). The Meta-analysis results showed the visual analog scale score of the bupivacaine group was lower than that of the placebo control group (weighted mean difference:-1.39, 95%confidence interval:-2.17 to-0.61, Pmean difference:157.72, 95%confidence interval:16.43 to 299.01, P<0.000 01). There was no significant difference in the incidence of side effect between two groups (relative risk:0.64, 95%confidence interval:0.29 to 1.44, P=0.48). On the basis of the currently available literature, the intra-articular of bupivacaine after arthroscopic surgery can significantly relieve pain without increasing the adverse reactions when compared with the placebo control group.