The association of rod curvature with postoperative outcomes in patients undergoing posterior lumbar interbody fusion for spinal stenosis: a retrospective case-control study.

BACKGROUND: Restoration of sagittal balance is a crucial consideration in posterior lumbar interbody fusion (PLIF) surgery and adverse postoperative outcomes are associated with inadequate restoration of sagittal alignment. However, there remains a shortage of substantial evidence regarding the effect of rod curvature on both sagittal spinopelvic radiographic parameters and clinical outcomes. METHOD: A retrospective case-control study was conducted in this study. Patient demographics (age, gender, height, weight and BMI), surgical characteristics (number of fused levels, surgical time, blood loss and hospital stay) and radiographic parameters (lumbar lordosis [LL], sacral slope [SS], pelvic incidence [PI], pelvic tilt [PT], PI-LL, Cobb angle of fused segments [Cobb], rod curvature [RC], Posterior tangent angle of fused segments [PTA] and RC-PTA) were analyzed. RESULTS: Patients in the abnormal group had older mean age and suffered more blood loss than those in the normal group. In addition, RC and RC-PTA were significantly lower in the abnormal group compared to the normal group. Multivariate regression analysis revealed that lower age (OR = 0.94; 95% CI: 0.89-0.99; P = 0.0187), lower PTA (OR = 0.91; 95% CI: 0.85-0.96; P = 0.0015) and higher RC (OR = 1.35; 95% CI: 1.20-1.51; P < 0.0001) were related to higher odds of better surgical outcomes. The receiver operating characteristic curve analysis showed that the ROC curve (AUC) for predicting outcomes of surgery by RC classifier was 0.851 (0.769-0.932). CONCLUSIONS: In patients who underwent PLIF surgery for lumbar spinal stenosis, those who had a satisfactory postoperative outcome tended to be younger, had lower blood loss, and higher values of RC and RC-PTA compared to those who had poor recovery and required revision surgery. Additionally, RC was found to be a reliable predictor of postoperative outcomes.

Long Non-coding RNA XIST May Influence Cervical Ossification of the Posterior Longitudinal Ligament Through Regulation of miR-17-5P/AHNAK/BMP2 Signaling Pathway.

Long non-coding RNAs (lncRNAs) play an important role in the development of bone-related diseases. This study was conducted to investigate the role and mechanism of lncRNA X inactive specific transcript (XIST) in the occurrence of cervical ossification of the posterior longitudinal ligament (OPLL). Here, primary human ligament fibroblasts cells (LFCs) were isolated from 30 cases of OPLL and 30 normal cervical posterior longitudinal ligament (non-OPLL) tissues to perform the qPCR and Western blot assay. We found that the mRNA level of lncRNA XIST was significantly increased in OPLL LFCs compared to non-OPLL LFCs. By bioinformatics analysis, we found that lncRNA XIST has four binding sites for miR-17-5p and found that the mRNA level of miR-17-5p was also significantly decreased in OPLL LFCs compared to non-OPLL LFCs. Since AHNAK is the target gene of miR-17-5p, we further found that the expression of AHNAK was significantly reduced in non-OPLL LFCs after being transfected with miR-17-5p mimic. The qPCR results showed that the mRNA expressions of BMP2 and Runx2 were significantly decreased. After being transfected with lncRNA XIST siRNA in the non-OPLL LFCs, the mRNA levels of lncRNA XIST, AHNAK, BMP2, and Runx2 were significantly decreased and the phosphorylated protein of Smad1/5/8 was reduced. After being cultured by mechanical vibration, the mRNA levels of lncRNA XIST, AHNAK, BMP2, Runx2, COL1, OC, OPN, and Phospho1 were significantly increased, but the mRNA expression of miR-17-5p was significantly decreased. The expression of phosphorylated Smad1/5/8 protein was also significantly increased. Together, this study was the first to determine that XIST gene inhibition plays an important role in the occurrence of cervical OPLL, through the mechanism of regulation of miR-17-5P/AHNAK/BMP2 signaling pathway. Thus, XIST may be a potential target that could be modulated for the treatment of cervical OPLL.

Proteomic analysis of the effects of cell culture density on the metastasis of breast cancer cells.

Cancer cell progression and proliferation increase cell density, resulting in changes to the tumour site, including the microenvironment. What is not known is if increased cell density influences the aggressiveness of cancer cells, especially their proliferation, migration, and invasion capabilities. In this study, we found that dense cell culture enhances the aggressiveness of the metastatic cancer cell lines, 4T1 and ZR-75-30, by increasing their proliferation, migration, and invasion capabilities. However, a less metastatic cell line, MCF-7, did not show an increase in aggressiveness, following dense cell culture conditions. We conducted a differential proteomic analysis on 4T1 cells cultured under dense or sparse conditions and identified an increase in expression for proteins involved in migration, including focal adhesion, cytoskeletal reorganization, and transendothelial migration. In contrast, 4T1 cells grown under sparse conditions had higher expression levels for proteins involved in metabolism, including lipid and phospholipid binding, lipid and cholesterol transporter activity, and protein binding. These results suggest that the high-density tumour microenvironment can cause a change in cellular behaviour, leading towards more aggressive cancers. SIGNIFICANCE OF THE STUDY: Metastasis of cancer cells is an obstacle to the clinical treatment of cancer. We found that dense cultures made metastatic cancer cells more potent in terms of proliferation, migration, and invasion. The proteomic and bioinformatic analyses provided some valuable clues for further intensive studies about the effects of cell density on cancer cell aggressiveness, which were associated with events such as pre-mRNA splicing and RNA transport, focal adhesion and cytoskeleton reorganization, ribosome biogenesis, and transendothelial migration, or associated with proteins, such as JAM-1 and S100A11. This investigation gives us new perspectives to investigate the metastasis mechanisms related to the microenvironment of tumour sites.

Anterior controllable antedisplacement fusion as a choice for 28 patients of cervical ossification of the posterior longitudinal ligament with dura ossification: the risk of cerebrospinal fluid leakage compared with anterior cervical corpectomy and fusion.

PURPOSE: To compare the incidence rate of cerebrospinal fluid (CSF) leakage between anterior controllable antedisplacement fusion (ACAF) and anterior cervical corpectomy and fusion (ACCF) in the treatment of ossification of the posterior longitudinal ligament (OPLL) with dura ossification (DO). METHODS: In the period from June 2015 to June 2017, ACAF and ACCF were performed on patients with OPLL with DO. Double-layer sign was observed on axial bone window of CT images. The operation duration, blood loss, and hospital stay were measured. Radiologic assessment included occupying rate, type and extent of OPLL, decompression width, postoperative area of the spinal canal, and anteroposterior diameter of the spinal cord. The JOA scoring system was used to evaluate the neurological status. Surgery-related complications such as CSF leakage and spinal cord or nerve injury were all recorded. RESULTS: There were 28 patients in ACAF group and 31 in ACCF group. On cross-sectional CT, decompression width and postoperative spinal canal area were both significantly larger in the ACAF group than that in the ACCF group (P < 0.01). The anteroposterior diameter of the spinal cord was significantly larger in the ACAF group (P < 0.05). Mean JOA score was better in the ACAF group (P < 0.05). In the ACCF group, seven (22.6%) patients had CSF leakage. However, only one (3.6%) presented with CSF leakage in the ACAF group. The difference of incidence rate of CSF leakage was significant (P < 0.01). CONCLUSIONS: ACAF, which can significantly reduce CSF leakage and achieve good neurological recovery, is a good option to treat cervical OPLL with DO. These slides can be retrieved under Electronic Supplementary Material.

Profiling and bioinformatics analysis of differentially expressed circular RNAs in human intervertebral disc degeneration.

The functional changes of nucleus pulposus (NP) cells are considered to be the initiating factors of intervertebral disc degeneration (IDD), and the differentially expressed circRNAs in NP cells may play an important role in the process of IDD. To identify circular RNAs (circRNAs) associated with human IDD, we isolated the NP cells from human degenerated and non-degenerated intervertebral disc and identified NP cells by microscopy and cell proliferation. CircRNA microarray expression profiles were obtained from NP cells of degenerated and non-degenerated intervertebral disc and further validated by quantitative reverse transcription PCR (qRT-PCR). The expression data were analyzed by bioinformatics. Microarray analysis identified 7294 circRNAs differentially expressed in degenerated human IDD NP cells. Among them, 3724 circRNAs were up-regulated and 3570 circRNAs were down-regulated by more than 2 folds. After validating by qRT-PCR, we predicted the possible miRNAs of the top dysregulated circRNAs using TargetScan, and miRanda. Furthermore, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that the most modulated circRNAs regulate the viability, degradation, apoptosis and oxidative stress in NP cells, and the possible mechanism underlying IDD was discussed. These results revealed that circRNAs may play a role in IDD and might be a promising candidate molecular target for gene therapy.

Skip corpectomy and fusion (SCF) versus anterior controllable antedisplacement and fusion (ACAF): which is better for patients with multilevel cervical OPLL?

BACKGROUND: The aim of this study was to compare the clinical efficacy and radiological outcomes of skip corpectomy and fusion (SCF) with anterior controllable antedisplacement and fusion (ACAF) for treating multilevel ossification of the posterior longitudinal ligament (OPLL). METHODS: 62 patients with multilevel OPLL who had undergone SCF or ACAF were analyzed retrospectively. Types of OPLL, occupying ratio (OR), Japanese Orthopaedic Association (JOA) score, Recovery Rate (RR), Neck Disability Index (NDI) score, Cobb's angles of C2-C7, operation time, blood loss, hospital stay, ratings for fusion assessment and complications were recorded and assessed. RESULTS: Postoperative C2-C7 Cobb's angle (11.1 ± 3.2° vs. 13.7 ± 2.5°; P < 0.05), NDI scores at final follow-up (14.3 ± 1.6 vs. 13.3 ± 1.3; P < 0.05), and rate of cerebral fluid (CSF) leakage (5, 16.7% vs. 0,0%; P < 0.05) were significantly better in the ACAF group. At 6 months, bone graft fusion rate was significantly greater in the ACAF group (24.75% vs. 15.50%; P < 0.05). CONCLUSIONS: Surgical treatment of multilevel OPLL by SCF or ACAF showed no significant differences in clinical outcomes, with the exception of better NDI scores at final follow-up in ACAF. In addition, ACAF is better than SCF in terms of early bone graft fusion rate, lordotic curvature improvement, risk of CSF leakage.

Anterior controllable antidisplacement and fusion surgery for the treatment of multilevel severe ossification of the posterior longitudinal ligament with myelopathy: preliminary clinical results of a novel technique.

PURPOSE: Although anterior and posterior decompression surgery are both reported to treat patients with myelopathy caused by ossification of the posterior longitudinal ligament (OPLL). The surgical strategy of the disease is still controversial when the OPLL is multilevel and severe. This present study reports the preliminary clinical results of a novel technique named anterior controllable antidisplacement and fusion (ACAF) for the treatment of multilevel-severe OPLL with myelopathy. METHODS: A series of 15 patients with cervical myelopathy caused by compression of multilevel severe OPLL were enrolled. All the patients underwent ACAF after thorough surgical designing based on preoperative imaging. The patients were followed for a mean follow-up duration of 9 months in this study. The main surgical procedures include discectomy of the involved levels, thinning of the anterior part of the involved vertebrae, intervertebral cages, anterior plate and screws installation, bilateral osteotomies of the vertebrae, and antedisplacement of the vertebrae-OPLL complex (VOC). The Japanese Orthopaedic Association (JOA) scales, Visual Analog Scale (VAS) were studied. And the pre- and postoperative radiological parameters, and surgical complications were also investigated. RESULTS: Postoperative CT and MRI showed complete decompression of the cord by antidisplacement of the VOC. Restoration of neurological defects was confirmed at the last follow-up assessment. Bone fusion was confirmed by CT at 6 months follow-up. No specific complications were identified that were associated with this technique. CONCLUSIONS: The present study demonstrates that excellent postoperative outcome can be achieved with the use of the ACAF. Though further study is required to confirm the conclusion, this novel technique has the potential to serve as an alternative surgical technique for the treatment of cervical OPLL. These slides can be retrieved under Electronic Supplementary Material.

Ossification of the posterior ligament is mediated by osterix via inhibition of the ß-catenin signaling pathway.

Ossification of the posterior longitudinal ligament (OPLL) involves ectopic calcification of the spinal ligament preferentially at the cervical spine. OPLL is associated with different diseases and occurs by endochondral ossification, which is associated with the activity of different transcription factors. However, the pathogenesis of OPLL remains unclear. Here, we investigated the role of osterix (Osx), a transcription factor that functions downstream of Runx2 and is an important regulator of osteogenesis, in the process of OPLL in a dexamethasone (Dex)-induced model of spinal ligament ossification. Our results showed that Osx is upregulated in patients with OPLL and during the ossification of ligament cells in parallel with the upregulation of osteogenic markers including osteocalcin (OCN), alkaline phosphatase (ALP) and collagen-1 (Col-1). Dex-induced ossification of ligament cells was associated with the downregulation and inactivation of ß-catenin, and these effects were offset by Osx knockdown. Activation of ß-catenin signaling abolished the effect of Dex on ossification and the upregulation of osteogenic markers. Taken together, our results suggest that OPLL is mediated by Osx via a mechanism involving the Wnt/ß-catenin signaling pathway, providing a basis for further research to identify potential targets for the treatment of OPLL.

Expansive open-door laminoplasty versus laminectomy and instrumented fusion for cases with cervical ossification of the posterior longitudinal ligament and straight lordosis.

PURPOSE: To identify whether expansive open-door laminoplasty (Lam) is more appropriate than laminectomy and instrumented fusion (LIF) for cases with ossification of the posterior longitudinal ligament (OPLL) and straight cervical lordosis. METHODS: A total of 67 cases were included and divided into Group Lam (n = 32) and Group LIF (n = 35), and the mean follow-up periods were 38 and 42 months, respectively. The cervical lordosis was elevated by C2-7 Cobb angle and cervical sagittal balance by C2-C7 sagittal vertical axis (SVA). Japanese Orthopedic Association (JOA), neurological recovery rate (RR) being calculated by the JOA, visual analog scale (VAS) and neck disability index (NDI) were used to assess clinical outcomes. RESULTS: Differences in general data between two groups were not significant. Total blood loss and operation duration in Group Lam were both significantly less than that in the Group LIF. By the final follow-up, the cervical lordosis significantly decreased in Group Lam and increased in Group LIF, the SVA significantly increased in Group Lam and kept unchanged in Group LIF, and the JOA, VAS, NDI significantly improved in both groups. Although there was no significant difference in RR between the two groups, cases in Group Lam had significantly larger incidence of postoperative kyphosis and kyphotic change rate, and less VAS, NDI and incidence of axial pain than cases in Group LIF. CONCLUSIONS: When compared with the LIF, the Lam is recommended for cases with OPLL and straight cervical lordosis when taking comparable neurological recovery, less axial pain and better neck function improvement into consideration.

The application of a new type of titanium mesh cage in hybrid anterior decompression and fusion technique for the treatment of continuously three-level cervical spondylotic myelopathy.

PURPOSE: To evaluate the efficacy and safety of a new type of titanium mesh cage (NTMC) in hybrid anterior decompression and fusion method (HDF) in treating continuously three-level cervical spondylotic myelopathy (TCSM). METHODS: Ninety-four cases who had TCSM and accepted the HDF from Jan 2007 to Jan 2010 were included. Clinical and radiological outcomes were compared between cases who had the NTMC (Group A, n = 45) and traditional titanium mesh cage (TTMC, Group B, n = 49) after corpectomies. Each case accepted one polyetheretherketone cage (PEEK) after discectomy. RESULTS: Mean follow-up were 74.4 and 77.3 months in Group A and B, respectively (p > 0.05). Differences in cervical lordosis (CL), segmental lordosis (SL), anterior segmental height (ASH) and posterior segmental height (PSH) between two groups were not significant preoperatively, 3-days postoperatively or at final visit. However, losses of the CL, SL, ASH and PSH were all significantly larger in Group B at the final visit, so did incidences of segmental subsidence and severe subsidence. Difference in preoperative Japanese Orthopedic Association (JOA), visual analog scale (VAS), neck disability index (NDI) or SF-36 between two groups was not significant. At the final visit, fusion rate, JOA, and SF-36 were all comparable between two groups, but the VAS and NDI were both significantly greater in Group B. CONCLUSIONS: For cases with TCSM, HDF with the NTMC and TTMC can provide comparable radiological and clinical improvements. But application of the NTMC in HDF is of advantages in decreasing the subsidence incidence, losses of lordosis correction, VAS and NDI.

Connexin 43 Affects Osteogenic Differentiation of the Posterior Longitudinal Ligament Cells via Regulation of ERK Activity by Stabilizing Runx2 in Ossification.

AIMS: Connexin 43 is one of the most potent gap junction proteins related to osteoblast differentiation and bone formation. We hypothesized that Connexin 43 is a significant factor in osteogenic differentiation in the posterior longitudinal ligament through the regulation of extracellular signal-regulated kinases (ERK) activity by converging on Runt-related transcription factor 2 (Runx2) activity. In this study, we mapped the activity of Connexin 43 to ERK and Runx2 by extracting longitudinal ligament cell for culture and silencing Connexin expression in addition to dexamethasone treatment in vitro. METHODS: qRT-PCR, Western Blot, and Runx2-responsive Luciferase Reporter Assay were performed to detect the activity of ERK, Runx2 and the expression levels of osseous genes under Connexin 43 modification. RESULTS: Downregulation of Connexin 43 resulted in suppression of dexamethasone-induced osteogenic differentiation, inhibition of the ERK and Runx2 activity, and reduction of osseous gene expression. CONCLUSION: these data support that Connexin 43 significantly regulates osteogenic differentiation in the cells from posterior longitudinal ligament by altering the activity of ERK, and subsequently causing the modification of Runx2.

MicroRNA-320a Regulates the Osteogenic Differentiation of Human Bone Marrow-Derived Mesenchymal Stem Cells by Targeting HOXA10.

BACKGROUND/AIMS: Human bone marrow-derived mesenchymal stem cells (hMSCs) are a promising cell source for bone engineering owing to their high potential to differentiate into osteoblasts. The bone morphogenetic protein-inducible gene homeobox a10 (HOXA10) is a critical regulator of osteogenesis. The objective of the present study was to identify microR-NAs (miRNAs) targeting HOXA10 and examine the effects on the osteogenic differentiation of hMSCs. METHODS: Based on in silico analysis, HOXA10-targeting miRNAs were selected and their regulatory roles in osteoblast differentiation were investigated. RESULTS: Six HOXA10-targeting miRNAs were identifIed by computational analysis, of which miR-320a was selected for further analysis because it was downregulated during osteogenic induction. Overexpression of miR-320a downregulated HOXA10 and significantly inhibited osteogenesis in hMSCs, as determined by the downregulation of the osteogenic markers Runx2, ALP, and OC and the inhibition of ALP activity and matrix mineralization, whereas miR-320a inhibition had the opposite effects. Furthermore, ectopic expression of HOXA10 (not including 3'-UTR) rescued the effects of miR-320a on osteogenic differentiation. CONCLUSION: These results suggest that miR-320a acts as a critical regulator of osteogenic differentiation of hMSCs by repressing its target HOXA10.

Connexin 43 promotes ossification of the posterior longitudinal ligament through activation of the ERK1/2 and p38 MAPK pathways.

Although cervical ossification of the posterior longitudinal ligament (OPLL) is one of the most common spinal diseases, the pathogenic mechanism is still not fully understood. Abnormal mechanical stress distribution is believed to be one of the main causes of OPLL. We have previously found that mechanical stress can up-regulate connexin 43 (Cx43) expression in ligament fibroblasts; this transduces mechanical signals to promote osteoblastic differentiation. In the present study, in order to explore further the intracellular mechanisms of Cx43-induced osteoblast differentiation of ligament fibroblasts, we investigate the potential roles of the osteogenic signaling pathway components ERK1/2, p38 MAPK and JNK in Cx43-mediated mechanical signal transduction. We first confirm higher Cx43 levels in both in vivo ligament tissue from OPLL patients and in vitro cultured OPLL cells. We find that ERK1/2, p38 MAPK and the JNK pathway are all activated both in vivo and in vitro. The activation of these signals was dependent upon Cx43, as its knock-down resulted in diminished mechanical effects and reduced signaling. Moreover, its knock-down almost reversed the osteogenic effect of mechanical stress on ligament fibroblasts and the blocking of the ERK1/2 and p38 MAPK pathways but not the JNK pathway, partly diminished this effect. Therefore, Cx43, which is up-regulated by mechanical stress, seems to function partly via the activation of ERK1/2 and p38 MAPK signals to promote the osteoblastic differentiation of ligament fibroblasts.

Implications of different patterns of "double-layer sign" in cervical ossification of the posterior longitudinal ligament.

PURPOSE: To make a preliminary classification of double-layer sign according to the morphological characteristics of the ossified and central hypodense mass and clarify implications of different patterns of "double-layer sign". METHODS: The 268 patients of cervical ossification of the posterior longitudinal ligament (OPLL) who underwent anterior corpectomy were retrospectively analyzed from January 2009 to January 2014. All these patients were performed cervical plain X-rays, CT and MRI. The double-layer sign was observed on axial bone window of CT images. According to the morphological characteristics of the ossified and central hypodense mass, this sign was classified into three types: type A was crescent shape, type B was short-straight shape and type C was long-straight shape. Type A was named when the central hypodense mass traced an arc and the OPLL is much more extensive than the dural ossification (DO). It belonged to type B when the central hypodense mass traced a short-straight line, less than or equal to half of the base width of the vertebrae. The OPLL may be extensive or equal to the DO. Type C was defined when it was more than half of the base width, presenting with a long-straight line and DO is much more extensive than OPLL. Intraoperative findings including dural mater ossification and adhesion, postoperative CSF leakage and outcome were all studied. Two spinal surgeons with rich experiences read the CT images according to this classification method and verified its consistency. RESULTS: Ninety-two patients were found in association with DO during the anterior decompression procedure, meanwhile the double-layer sign could be seen on axial bone window of CT imaging. In these 92 patients, there were 51 patients presenting with type A, and the ossified mass was completely resected with ossified dura mater reserved. Only two patients presented with dural defect and postoperative CSF leakage. Thirty-five patients was classified as type B, of which six accompanied by CSF leakage. All the other six patients of type C presented with CSF leakage after operation. There was a significant correlation between the occurence of CSF leakage and pattern of double-layer sign, but not gender, age, duration of symptoms, extent of OPLL and occupying rate. The Kappa value between the two surgeons was 0.82, showing a good consistency of the method. CONCLUSIONS: OPLL patients with double-layer sign of type C is almost inevitably followed by CSF leakage after anterior decompression. For type A and B, occurrence of CSF leakage is not as high as we thought before. Pattern of double-layer sign should be a considered factor when anterior or posterior approach is chosen.

A new method to determine whether ossified posterior longitudinal ligament can be resected completely and safely: spinal canal "Rule of Nine" on axial computed tomography.

PURPOSE: To introduce the new method of spinal canal "Rule of Nine" on axial computed tomography (CT) to determine whether ossified posterior longitudinal ligament (OPLL) can be resected completely and safely. METHODS: From January 2008 to January 2013, we retrospectively analyzed 308 patients with cervical OPLL who underwent anterior corpectomy. Patients were examined by cervical plain X-ray, CT, and magnetic resonance imaging. The characteristics of OPLL on axial CT, dural mater adhesion and ossification, complete resection of the ossified mass, cerebrospinal fluid (CSF) leakage, spinal cord injury (SCI), and postoperative outcome were all studied. OPLL was characterized at the maximum ossified level using spinal canal "Rule of Nine" on axial CT. RESULTS: OPLL was completely and safely resected in 129 patients, whose ossified mass was within the safety zone. No CSF leakage or SCI occurred. The most ossified material was resected when OPLL was within the danger zone in 21 patients. There were two cases of SCI and seven of CSF leakage. In the other 158 patients, the ossified mass was within the intermediate zone. In 96 patients the width of the ossified mass was <2 resection units. It was completely resected and one case had CSF leakage. In the other 62 cases, the ossified mass was >2 resection units and there were nine cases of CSF leakage. CSF leakage and SCI increased significantly when the ossified mass was ≥6 units. The inter- and intraobserver κ values were 0.76 and 0.84, respectively, showing a good reliability. CONCLUSIONS: Our new method can be used to determine whether OPLL can be resected completely and safely, and identify potential risks and complications.

Metabolic profiling study on potential toxicity and immunotoxicity-biomarker discovery in rats treated with cyclophosphamide using HPLC-ESI-IT-TOF-MS.

Despite the recent advances in understanding toxicity mechanism of cyclophosphamide (CTX), the development of biomarkers is still essential. CTX-induced immunotoxicity in rats by a metabonomics approach was investigated using high-performance liquid chromatography coupled with ion trap time-of-flight mass spectrometry (HPLC-ESI-IT-TOF-MS). The rats were orally administered CTX (30 mg/kg/day) for five consecutive days, and on the fifth day samples of urine, thymus and spleen were collected and analyzed. A significant difference in metabolic profiling was observed between the CTX-treated group and the control group by partial least squares-discriminant analysis (PLS-DA), which indicated that metabolic disturbances of immunotoxicity in CTX-treated rats had occurred. One potential biomarker in spleen, three in urine and three in thymus were identified. It is suggested that the CTX-toxicity mechanism may involve the modulation of tryptophan metabolism, phospholipid metabolism and energy metabolism. This research can help to elucidate the CTX-influenced pathways at a low dose and can further help to indicate the patients' pathological status at earlier stages of toxicological progression after drug administration.

Zero-profile integrated plate and spacer device reduces rate of adjacent-level ossification development and dysphagia compared to ACDF with plating and cage system.

STUDY DESIGN: Retrospective case-control study. PURPOSE: To compare the safety and efficacy of the Zero-profile (Zero-p) integrated plate and spacer device to that of an anterior cervical plate and cage in patients undergoing anterior cervical discectomy and fusion (ACDF). Anterior cervical plating system has provided good results, including higher fusion rate and improved alignment since its use. However, adjacent-level ossification development (ALOD) and dysphagia have been usually reported associating with plates. METHODS: This was a retrospective control study. Sixty-two patients with cervical radiculopathy or myelopathy were treated with an anterior plate and cage or Zero-p implant between January 2011 and December 2011. The mean follow-up was 33.1 months in the plate and cage group and 30.6 months in Zero-p group. Patient demographics, operative details and complications were reviewed. The clinical outcomes were evaluated using the Japanese Orthopaedic Association (JOA) scores and JOA recovery rate before and after operations. Incidence of cephalad and caudal ALOD on the lateral radiographs was studied at preoperation, immediate postoperation and last follow-up. Incidence of dysphagia was also recorded after operation according to Bazaz-Yoo dysphagia index. RESULTS: Thirty-two patients received an anterior plate and cage and 30 received the Zero-p implant. There were no statistical differences in patient demographics, operative details between the two groups. The JOA scores significantly increased compared with preoperational measurements in both groups (p < 0.05), but the JOA recovery rate was similar (72.2 % for plate and cage group and 77.0 % for Zero-p group, p > 0.05). ALOD occurred in 12 (18.8 %) of the 64 cephalad and caudal adjacent segments in plate with cage group, and only 1 (1.6 %) of 63 adjacent levels (including three noncontiguous cases) presented with ALOD in Zero-p group. The difference was significant (p < 0.01). The incidence of dysphagia in the Zero-p group was lower compared with that in the plate with cage group, and the symptom duration was much shorter (p < 0.01). Both groups had no adverse events associated with the implant or implant surgery. CONCLUSIONS: The Zero-profile implant is safe and efficacious after ACDF. It can reduce the rate of adjacent-level ossification development and dysphagia compared to anterior plate and cage.

Upregulated expression of PERK in spinal ligament fibroblasts from the patients with ossification of the posterior longitudinal ligament.

PURPOSE: Molecular mechanism of ossification of the posterior longitudinal ligament (OPLL) remains unclear. This study was to investigate different expressions of PERK between the spinal ligament fibroblasts from OPLL patients and non-OPLL patients, and demonstrate knockdown of PERK protein expression by RNA interference inhibiting expression of osteocalcin (OCN), alkaline phosphatase (ALP), and type I collagen (COL I) in the cells from OPLL patients. METHODS: Spinal ligament cells were cultured using tissue fragment cell culture and identified by immunocytochemistry and immunofluorescence. The mRNA expression of osteoblast-specific genes of OCN, ALP and COL I was detected in the cells from OPLL and non-OPLL patients by semiquantitative reverse transcription-polymerase chain reaction. The protein expression of PERK was detected by Western blotting. And then, after 72 h, when RNA interference against PERK was performed on the cells from OPLL patients, expression of the osteoblast-specific genes was compared again between the transfection group and non-transfection group. RESULTS: Spinal ligament fibroblasts were observed 7-10 days after cell culture. Immunocytochemistry and immunofluorescence exhibited positive results of vimentin staining. The mRNA expressions of OCN, ALP and COL I and protein expression of PERK in the cells from OPLL patients were significantly greater than those from non-OPLL patients. In addition, knockdown of PERK protein expression inhibited the mRNA expressions of OCN, ALP and COL I remarkably in the transfection group compared with the non-transfection group, at 72 h after RNA interference targeting PERK was performed on the cells from OPLL patients. CONCLUSIONS: The cultured fibroblasts from OPLL patients exhibited osteogenic characteristics, and PERK-mediated ER stress might be involved in development of OPLL.