Si-Dong Yang

17β-Estradiol protects against apoptosis induced by levofloxacin in rat nucleus pulposus cells by upregulating integrin α2β1

Levofloxacin has been reported to have cytotoxicity to chondrocytes in vitro. And 17β-estradiol has been widely studied for its protective effects against cell apoptosis. Based on apoptotic cell model induced by levofloxacin, the purpose of this study was to explore the mechanism by which 17β-estradiol protects rat nucleus pulposus cells from apoptosis. Inverted phase-contrast microscopy, flow cytometry, and caspase-3 activity assay were used to find that levofloxacin induced marked apoptosis, which was abolished by 17β-estradiol. Interestingly, estrogen receptor antagonist, ICI182780, and functional blocking antibody to α2β1 integrin, both prohibited the effect of 17β-estradiol. Simultaneously, levofloxacin decreased cellular binding ability to type II collagen, which was also reversed by 17β-estradiol. Furthermore, western blot and real-time quantitative PCR were used to find that integrin α2β1 was responsible for estrogen-dependent anti-apoptosis, which was time–response and dose–response effect. 17β-estradiol was proved for the first time to protect rat nucleus pulposus cells against levofloxacin-induced apoptosis by upregulating integrin α2β1 signal pathway.

Combined effect of 17β-estradiol and resveratrol against apoptosis induced by interleukin-1β in rat nucleus pulposus cells via PI3K/Akt/caspase-3 pathway

Background: In previous studies, both 17β-estradiol (E2) and resveratrol (RES) were reported to protect intervertebral disc cells against aberrant apoptosis. Given that E2 has a better anti-apoptotic effect with more cancer risk and RES has an anti-apoptotic effect with less cancer risk, the combined use of E2 with RES is promising in developing clinical therapies to treat apoptosis-related diseases such as intervertebral disc degeneration in the future. Objective: The purpose of this study was to explore the combined effect of E2 with RES on rat nucleus pulposus cells and the underlying mechanisms. Methods: TUNEL assay and FACS analysis were used to determine apoptotic incidence of nucleus pulposus cells. MTS assay was used to determine cell viability, and cellular binding assay was used to determine cell-ECM (extracellular matrix) ability. Real-time quantitative RT-PCR was to determine mRNA level of target genes. And Western blot was used to determine the protein level. Results: Both E2 and RES decreased apoptotic incidence when used singly; interestingly, they decreased apoptosis more efficiently when used combinedly. Meanwhile, E2 and RES combined together against the decrease of cell viability and binding ability resulting from IL-1β cytotoxicity. As well, activated caspase-3 was suppressed by the combined effect. Furthermore, IL-1β downregulated expression level of type II collagen and aggrecan (standing for anabolism), while upregulated MMP-3 and MMP-13 (standing for catabolism). However, the combined use of E2 with RES effectively abolished the above negative effects caused by IL-1β, better than either single use. Finally, it turned out to be that E2 and RES combined together against apoptosis via the activation of PI3K/Akt/caspase-3 pathway. Conclusion: This study presented that IL-1β induced aberrant apoptosis, which was efficiently resisted by the combined use of E2 with RES via PI3K/Akt/caspase-3 pathway.

Prevalence and risk factors of deep vein thrombosis in patients after spine surgery: a retrospective case-cohort study

Deep vein thrombosis (DVT) is common seen in patients undergoing spine surgery. However, its prevalence and associated risk factors have not been well understood yet. This retrospective case-cohort study was designed to investigate risk factors for postoperative DVT using retrospectively collected data from department of spine surgery between 07/2013 and 07/2014. Univariate analysis and binary logistic regression analysis were used to determine risk factors for DVT. A total of 861 patients were admitted into DVT-associated analyses, including 410 males and 451 females, aged from 15 to 87 years old (median 54, IQR 18). Of them, 147 cases (17%) sustained postoperative DVT. DVT incidence was 15.9% in patients undergoing lumbar interbody fusion, 13.5% in patients treated by low-molecular-weight heparin (LMWH), while only 8.1% in patients without LMWH. However, it revealed no significant difference between LMWH group and non-LMWH group (χ2 = 1.933, p = 0.164). Logistic regression equation was logit P = −4.09 + 0.05*X1 − 0.55*X2 + 0.41*X3 + 1.41*X7, (X1 = age; X2 = regions; X3 = hypertension; X7 = D-dimer). In this study, LMWH prophylaxis after spine surgery proved ineffective. Advanced age, D-dimer and hypertension have proved to be the risk factors for postoperative DVT in patients undergoing spine surgery.

17β-Estradiol protects against apoptosis induced by interleukin-1β in rat nucleus pulposus cells by down-regulating MMP-3 and MMP-13

In our previous study, 17β-estradiol was proved to protect rat annulus fibrosus cells against apoptosis induced by interleukin-1β (IL-1β). However, whether 17β-estradiol has protective effect on rat nucleus pulposus cells remains unclear. The purpose of this study was to further explore the effects of 17β-estradiol on rat nucleus pulposus cells based on IL-1β-induced apoptosis. TUNEL assay and Annexin V/PI double staining were used to detect apoptosis and revealed that IL-1β induced notable apoptosis, which was reversed by 17β-estradiol. Meanwhile, cell viability and binding ability were decreased by IL-1β, but activated caspase-3 was increased. However, all of the detected effects of IL-1β were eliminated by 17β-estradiol. Furthermore, real-time quantitative RT-PCR was used to further find that IL-1β downregulated expression level of type II collagen, aggrecan, tissue inhibitor of matrix metalloproteinase (TIMP)-1, while upregulated matrix metalloproteinase (MMP)-3, MMP-13 and Bcl-2, which was further confirmed by western blot. Finally, 17β-estradiol was proved to abolish the above negative effects of IL-1β. In summary, this work presented that IL-1β maybe induced apoptosis of rat nucleus pulposus cells, which was resisted by 17β-estradiol by down-regulating MMP-3 and MMP-13 via a mitochondrial pathway. This research provides a novel insight into the anti-apoptotic effect of 17β-estradiol on IL-1β-induced cytotoxicity, and may potentially lead to a better understanding of the clinical effects of 17β-estradiol, especially in terms of intervertebral disc degeneration.

Levofloxacin increases the effect of serum deprivation on anoikis of rat nucleus pulposus cells via Bax/Bcl-2/caspase-3 pathway

Abstract Levofloxacin, a fluoroquinolone, is a widely-used and effective antibiotic. However, various adverse side effects are associated with levofloxacin. The purpose of this study was to further explore the effects of levofloxacin on rat nucleus pulposus cells (NPCs). Inverted phase-contrast microscopy, flow cytometry and caspase-3 activity assays were used and revealed that serum deprivation induced apoptosis, which was markedly increased by levofloxacin in a dose-dependent manner. Simultaneously, levofloxacin decreased cell binding to type II collagen (COL2). Thus, levofloxacin-induced apoptosis exhibits characteristics of anoikis, the process by which cell death is triggered by separation from the extracellular matrix, which contains COL2. Furthermore, real-time quantitative RT-PCR was used to further confirm that levofloxacin downregulates COL2 expression in a dose-dependent manner. At last, western blot was used to find that levofloxacin increased the ratio of Bax/Bcl-2 and active caspase-3 in a dose-dependent manner. Levofloxacin therefore increases the effects of serum deprivation on anoikis by downregulating COL2 in rat NPCs in vitro via Bax/Bcl-2/caspase-3 pathway. This research provides a novel insight into the mechanisms of levofloxacin-induced toxicity and may potentially lead to a better understanding of the clinical effects of levofloxacin, especially in terms of intervertebral disc degeneration.

Incidence and risk factors of adjacent segment disease following posterior decompression and instrumented fusion for degenerative lumbar disorders

Abstract The purpose of this study was to explore incidence and risk factors of adjacent segment disease (ASD) following posterior decompression and instrumented fusion for degenerative lumbar disorders, and hope to provide references in decision making and surgical planning for both spinal surgeon and surgically treated patients. By retrieving the medical records from January 2011 to December 2013 in our hospital, 237 patients were retrospectively reviewed. According to the occurrence of ASD at follow up, patients were divided into 2 groups: ASD and N-ASD group. To investigate risk values for the occurrence of ASD, 3 categorized factors were analyzed statistically: Patient characteristics: age, sex, body mass index (BMI), bone mineral density (BMD), duration. Surgical variables: surgical strategy, number of fusion level, surgery segment, surgery time, blood loss, intraoperative superior facet joint violation. Radiographic parameters: preoperative lumbar lordosis, preoperative angular motion at adjacent segment, preoperative adjacent segment disc degeneration, preoperative paraspinal muscle degeneration. Postoperative ASD was developed in 15 of 237 patients (6.3%) at final follow up. There was no statistically significant difference between the 2 groups in patient characteristics of age, sex composition, BMD, duration, while the BMI was higher in ASD group than that in N-ASD group. There was no difference in surgical variables of surgical strategy, number of fusion level, surgery segment, surgery time, blood loss, while intraoperative superior facet joint violation was more common in ASD group than that in N-ASD group. There was no difference in radiographic parameters of preoperative lumbar lordosis, preoperative paraspinal muscle degeneration, while preoperative adjacent segment disc degeneration were more severe in ASD group than that in N-ASD group. The Logistic regression analysis revealed that, BMI >25 kg/m2, preoperative disc degeneration, and superior facet joint violation were independently associated with ASD. In conclusion, higher BMI, preoperative disc degeneration at adjacent segment and intraoperative superior facet joint violation are risk factors for ASD. Patients who are overweight or obesity and with preoperative disc degeneration at adjacent segment should be fully informed the risk of ASD. For surgeons, it is essential to prevent superior facet joint violation in pedicle screw insertion procedure.

Incidence and risk factors for the progression of proximal junctional kyphosis in degenerative lumbar scoliosis following long instrumented posterior spinal fusion

AbstractThe aim of this study was to identify the prevalence of proximal junctional kyphosis (PJK) in degenerative lumbar scoliosis (DLS) following long instrumented posterior spinal fusion, and to search for predictable risk factors for the progression of junctional kyphosis.In total 98 DLS patients with a minimum 2-year follow-up were reviewed prospectively. According to the occurrence of PJK at the last follow-up, patients were divided into 2 groups: PJK group and non-PJK group. To investigate risk values for the progression of PJK, 3 categorized factors were analyzed statistically: patient characteristics—preoperative data of age, sex, body mass index (BMI), bone mineral density (BMD) were investigated; surgical variables—the most proximal and distal levels of the instrumentation, the number of instrumented levels; pre- and postoperative radiographic parameters include the scoliotic angle, sagittal vertical axis, thoracic kyphosis, thoracolumbar junctional angle, lumbar lordosis, pelvic incidence, pelvic tilt, and sacral slope.PJK was developed in 17 of 98 patients (17.3%) until to the final follow-up and were enrolled as the PJK group, and 81 patients without PJK at final follow-up were enrolled as the non-PJK group. There was no statistically significant difference between the 2 groups in age at operation (P = 0.900). The patients sex was excluded in statistical analysis because of the predominance of female patients. There were statistically significant difference between the 2 groups in BMI ([25.5 ± 1.7] kg/m2 in the PJK group and [23.6 ± 1.9] kg/m2 in the non-PJK group, P < 0.001) and BMD ([–1.4 ± 0.8] g/cm2 in the PJK group and [−0.7 ± 0.3] g/cm2 in the non-PJK group, P < 0.001). No specific surgery-related variables were found to be associated with an increased risk of developing PJK, except when the most proximal instrumented vertebrae stopped at thoracolumbar junction (T11-L1). The upper instrumentation vertebrae (UIV) at thoracolumbar junction was more common in the PJK group than that in the non-PJK group (P = 0.007). No preoperative and early postoperative variable did reveal a statistically significant difference between the 2 groups. When included in a multivariate logistic regression model, BMI>25 kg/m2, osteoporosis, and UIV at thoracolumbar junction were independently associated with PJK.In conclusion, osteoporosis, obesity, and UIV at thoracolumbar junction are risk factors for the development and progression of PJK in DLS patients following long instrumented posterior spinal fusion. Antiosteoporosis treatment extends the fusion level above the thoracolumbar region and controlling body weight before and after surgery could provide opportunities to reduce the rate of PJK and to improve therapeutic outcomes.

Laminoplasty versus laminectomy and fusion for multilevel cervical compressive myelopathy: A meta-analysis.

AbstractThis is a meta-analysis to compare the results between laminoplasty and laminectomy followed by fusion for the patients with multilevel cervical compressive myelopathy. An extensive search of literature was performed in MEDLINE, Embase, the Cochrane library, CNKI, and WANFANG. The following outcome measures were extracted: the Japanese Orthopaedic Association (JOA) scores, cervical curvature index (CCI), visual analog scale (VAS), cervical lordosis (C2–7), complications, blood loss, and operation time. Data analysis was conducted with RevMan 5.3 and STATA 12.0. A total of 23 studies comprising 774 and 743 patients treated with laminoplasty and laminectomy followed by fusion, respectively, were included in the final analysis. The pooled analysis showed that there was no significant difference in preoperative JOA scores [P = 0.89], postoperative JOA scores [P = 0.13], JOA scores improvement rate [P = 0.27], preoperative CCI [P = 0.15], postoperative CCI [P = 0.14], preoperative VAS [P = 0.41], postoperative VAS [P = 0.52], preoperative cervical lordosis (C2–7) [P = 0.46], postoperative cervical lordosis (C2–7) [P = 0.67], total complications [P = 0.07], axial pain [P = 0.94], and blood loss [P = 0.51]. However, there were significant difference in operation time (WMD = −19.57 [−32.11, −7.02], P = 0.002) and C5 palsy (OR = 0.26 [0.15, 0.44], P < 0.001). As compared with laminectomy followed by fusion, expansive laminoplasty showed no significant differences in JOA scores, CCI, ROM, VAS, cervical lordosis (C2–7), axial pain, total complications, and blood loss, but shorter operation time and fewer C5 palsy.

Prevalence and Risk Factors of Deep Vein Thrombosis in Patients Undergoing Lumbar Interbody Fusion Surgery: A Single-Center Cross-Sectional Study.

AbstractThis cross-sectional study was designed to obtain the current prevalence of deep vein thrombosis (DVT) and analyze related risk factors in patients undergoing lumbar interbody fusion.Medical record data were collected from Department of Spinal Surgery, The Third Hospital of Hebei Medical University, between July 2014 and March 2015. Both univariate analysis and binary logistic regression analysis were performed to determine risk factors for DVT.A total of 995 patients were admitted into this study, including 484 men and 511 women, aged from 14 to 89 years old (median 50, IQR 19). The detection rate of lower limb DVT by ultrasonography was 22.4% (223/995) in patients undergoing lumbar interbody fusion. Notably, average VAS (visual analog scale) score in the first 3 days after surgery in the DVT group was more than that in the non-DVT group (Z = −21.69, P < 0.001). The logistic regression model was established as logit P = −13.257 + 0.056*X1 − 0.243*X8 + 2.085*X10 + 0.001*X12, (X1 = age; X8 = HDL; X10 = VAS; X12 = blood transfusion; x2 = 677.763, P < 0.001).In conclusion, advanced age, high postoperative VAS scores, and blood transfusion were risk factors for postoperative lower limb DVT. As well, the logistic regression model may contribute to an early evaluation postoperatively to ascertain the risk of lower limb DVT in patients undergoing lumbar interbody fusion surgery.

The incidence and risk factors of postoperative neurological deterioration after posterior decompression with or without instrumented fusion for thoracic myelopathy.

Abstract The aim of this study was to explore the incidence and risk factors of postoperative neurological deterioration after posterior decompression with or without instrumented fusion for thoracic myelopathy, and hope to provide references in decision-making and surgical planning for both spinal surgeon and thoracic stenosis patients. By retrieving the medical records from January 2001 to November 2015, 168 patients were retrospectively reviewed. According to the occurrence of postoperative neurological deterioration, patients were divided into 2 groups: neurological deterioration (ND) group and non-ND group. To investigate risk values for the occurrence of ND, 3 categorized factors were analyzed statistically: patient characteristics—preoperative data of age, sex, body mass index, bone mineral density, the duration of disease (from first symptoms to operation), the preoperative neurological function (Frankel grade), and diagnosis; surgical variables—surgery time, the amount of bleeding, mean arterial pressure, intervertebral fusion or not, and instrumentation or not; radiographic parameters—the spinal canal occupancy ratio, location of the lesion, thoracic kyphosis, and kyphosis correction. Postoperative neurological deterioration was developed in 23 of 168 patients (13.7%), and were enrolled as ND group. There was no statistically significant difference between the 2 groups in age at operation, sex composition, body mass index, and bone mineral density. The preoperative diagnosis presented significant difference between the 2 groups, because ossification of posterior longitudinal ligament combined with ossification of the ligamentum flavum was more common in ND group, whereas ossification of the ligamentum flavum alone was more common in non-ND group. There was no difference between the 2 groups in mean surgery time, the incidence of intraoperative direct trauma, and the number of patients that received instrumentation. The mean bleeding was much more in ND group than that in non-ND group, and the mean arterial pressure was lower in ND group than that in non-ND group. Also, the mean spinal canal occupancy ratio was more severe in ND group than that in non-ND group. There were no statistically significant difference between the 2 groups in stenosis location and preoperative thoracic kyphosis. The mean kyphosis correction was more significant in ND group. When included in a multivariate logistic regression model, thoracic disc herniation + ossification of posterior longitudinal ligament, spinal canal occupancy ratio more than 70%, bleeding more than 800 mL, and mean arterial pressure less than 81 mm Hg were independently associated with the postoperative neurological deterioration. In conclusion, ossification of posterior longitudinal ligament combined with ossification of the ligamentum flavum, spinal canal occupancy ratio more than 70%, intraoperative bleeding more than 800 mL, and mean arterial pressure less than 81 mm Hg are risk factors for the occurrence of postoperative neurologic deterioration. Improving surgical technique, shortening operation time, and paying more attention to hemostasis could provide opportunities to reduce the incidence of neurologic deterioration and to improve therapeutic outcomes.