Development and validation of an LC-MS/MS method for the determination of hinokiflavone in rat plasma and its application to a pharmacokinetic study.

Hinokiflavone has drawn a lot of attention for its multiple biological activities. In this study, a sensitive and selective method for determination of hinokiflavone in rat plasma was developed for the first time, using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Amentoflavone was used as an internal standard. Separation was achieved on a Hypersil Gold C18 column with isocratic elution using methanol-water (65:35, v/v) as mobile phase at a flow rate of 0.3 mL/min. A triple quadrupole mass spectrometer operating in the negative electrospray mode with selected reaction monitoring was used to detect the transitions of m/z 537 → 284 for hinokiflavone and m/z 537 → 375 for IS. The LOQ was 0.9 ng/mL with a linear range of 0.9-1000 ng/mL. The intra- and inter-day accuracy (RE%) ranged from -3.75 to 6.91% and from -9.20 to 2.51% and the intra- and inter-day precision (RSD) was between 0.32-14.11 and 2.85-10.04%. The validated assay was successfully applied to a pharmacokinetic study of hinokiflavone in rats. The half-life of drug elimination at the terminal phase was 6.10 ± 1.86 h, and the area under the plasma concentration-time curve from time zero to the time of last measurable concentration and to infinity values obtained were 2394.42 ± 466.86 and 2541.93 ± 529.85 h ng/mL, respectively.

Correlation of bone morphogenetic protein-2 levels in serum and synovial fluid with disease severity of knee osteoarthritis.

BACKGROUND: This study aimed to investigate the bone morphogenetic protein-2 (BMP-2) levels in serum and synovial fluid (SF) of patients with primary knee osteoarthritis (OA) and to exam its correlation with radiographic and symptomatic severity of the disease. MATERIAL/METHODS: A total of 37 knee OA patients and 20 healthy controls were enrolled in this study. Knee OA radiographic grading was performed according to the Kellgren-Lawrence (KL) grading system by evaluating X-ray changes observed in anteroposterior knee radiography. Symptomatic severity of the disease was evaluated according to the Western Ontario McMaster University Osteoarthritis Index (WOMAC) scores. BMP-2 levels in serum and SF were determined using enzyme-linked immunosorbent assay. RESULTS: Serum BMP-2 level in patients with knee OA was higher than that in healthy controls. Knee OA patients with KL grade 4 showed significantly elevated BMP-2 levels in the serum and SF compared with those with KL grade 2 and 3. Knee OA patients with KL grade 3 had significant higher SF levels of BMP-2 than those with KL grade 2. BMP-2 levels in the serum and SF of knee OA patients were both positively correlated with KL grades and WOMAC scores. CONCLUSIONS: BMP2 levels in serum and SF were closely related to the radiographic and symptomatic severity of knee OA and may serve as an alternative biochemical parameter to determine disease severity of primary knee OA.

[Histocompatibility of nano-hydroxyapatite/poly-co-glycolic acid tissue engineering bone modified by mesenchymal stem cells with vascular endothelial frowth factor].

OBJECTIVE: To explorec Histocompatibility of nano-hydroxyapatite/poly-co-glycolic acid tissue engineering bone modified by mesenchymal stem cells with vascular endothelial frowth factor transinfected. METHODS: Rat bone marrow mesenchymal stem cells (BMSCs) was separated, using BMSCs as target cells, and then vascular endothelial growth factor (VEGF) gene was transfected. Composite bone marrow mesenchymal stem cells and cells transfected with nano-hydroxyapatite (HA)/polylactic-co-glycolic acid (PLGA). The composition of cell and scaffold was observed. RESULTS: The blank plasmid transfection was 39.1%, 40.1% in VEGF group. The cell adhesion and growth was found on the scaffold pore wall after 5 days, and the number of adherent cells in the nano-HA/PLGA composite scaffold material basically had no significant difference in both. CONCLUSION: Although the nano-HA/PLGA scaffold material is still not fully meet the requirements of the matrix material for bone tissue engineering, but good biocompatibility, structure is its rich microporous satisfaction in material mechanics, toughening, enhanced obviously. Composition scaffold with BMSCs transfected by VEGF plasmid, the ability of angiogenesis is promoted.

[Distribution of Schmorl's nodes in lumbar spine and their relationship with lumbar disc degeneration].

OBJECTIVE: To explore the distribution of Schmorl's nodes (SN) in normal adult lumbar spine and determine the association of lumbar disc degeneration and lumbar spine motion. METHODS: A total of 1 179 individuals underwent magnetic resonance imaging (MRI) scans. They had an age range of 15-85 years and were grouped by 10 years. The distribution of SNs along lumbar spine and their relationship with age and gender were examined by single factor analysis of variance chi-square test. Spine specialists performed lumbar disc degeneration evaluations through the Pfirrmann's classification system. The relationship between SNs, age group, disc location and overall grades of lumbar disc degeneration were analyzed by multiple Logistic regression. RESULTS: The prevalence of SNs was 28.4%. And SNs were observed more frequently in males than females (34.6% vs 20.2%) (P < 0.01). No significant inter-group difference existed in the incidence of SNs (P = 0.18). SNs were more common at L2 and L3 vertebral bodies (14.3% vs 14.4%) while least common at S1 vertebral body (1.5%). The highest incidence of SNs was found in discs with degeneration grades III (41.9%) and IV (45.3%). The occurrence of SN, aging and disc location were positively correlated with lumbar disc degeneration grade. CONCLUSION: With a high incidence in healthy individuals, SNs occur in discs at all degrees of degeneration. And the occurrence of SNs is positively associated with lumbar disc degeneration.

LncRNA MALAT1 mediates osteogenic differentiation in osteoporosis by regulating the miR-485-5p/WNT7B axis.

Introduction: Accumulating evidence demonstrates that long non-coding RNAs (lncRNAs) are associated with the development of osteoporosis. Methods: This study aimed to investigate the effects of MALAT1 on osteogenic differentiation and cell apoptosis in osteoporosis. MALAT1 level, detected by RT-qPCR, was downregulated in hindlimb unloading (HU) mice and simulated microgravity (MG)-treated MC3T3-E1 cells. Moreover, osteogenic differentiation-related factor (Bmp4, Col1a1, and Spp1) levels were measured by RT-qPCR and Western blot. ALP activity was detected, and ALP staining was performed. Cell apoptosis was assessed by flow cytometry. Results: The results revealed that MALAT1 upregulated the expression of Bmp4, Col1a1, and Spp1, and enhanced ALP activity. Knockdown of MALAT1 suppressed their expression and ALP activity, suggesting that MALAT1 promoted osteogenic differentiation. Additionally, MALAT1 inhibited apoptosis, increased Bax and caspase-3 levels, and decreased Bcl-2 level. However, knockdown of MALAT1 had opposite results. In MG cells, MALAT1 facilitated osteogenic differentiation and suppressed apoptosis. Furthermore, miR-485-5p was identified as a target of MALAT1, and WNT7B was verified as a target of miR-485-5p. Overexpression of miR-485-5p rescued the promotion of osteogenic differentiation and the inhibition of apoptosis induced by MALAT1. Knockdown of WNT7B abolished the facilitation of osteogenic differentiation and the suppression of apoptosis induced by downregulation of miR-485-5p. Discussion: In conclusion, MALAT1 promoted osteogenic differentiation and inhibited cell apoptosis through the miR-485-5p/WNT7B axis, which suggested that MALAT1 is a potential target to alleviate osteoporosis.

LncRNA MIAT can regulate the proliferation, apoptosis, and osteogenic differentiation of bone marrow-derived mesenchymal stem cells by targeting miR-150-5p.

Osteoporosis (OP) is a systemic bone metabolic disease with complicated pathogenesis and is difficult to cure clinically. The regulatory mechanisms of OP are needed to be further investigated. In the present study, we focused on the role of myocardial infarction-associated transcript (MIAT) in OP development and examined the underlying mechanism. The serum expression levels of MIAT in samples from patients with OP and healthy controls were compared using quantitative reverse transcription-PCR (qRT-PCR). The dual-luciferase reporter assay was used to confirm the relationship between MIAT and its potential target microRNA, i.e., miR-150-5p. Moreover, bone marrow-derived mesenchymal stem cells (BMSCs) were cultured and transfected with MIAT shRNA, with or without miR-150-5p inhibitor. EdU staining and colony formation analysis were performed to determine the proliferation ability of these cells. Furthermore, the TUNEL assay and flow cytometry were used to assess BMSC apoptosis. Finally, RT-PCR and Western blot assays were employed to assess the expression of osteogenic differentiation biomarkers. Compared with controls, the expression of MIAT was significantly increased, whereas that of miR-150-5p was markedly decreased in patients with OP. MIAT and miR-150-5p expression levels exhibited a strong negative correlation. Furthermore, osteogenic differentiation indicators were suppressed in serum of OP patients. MIAT was downregulated, and miR-150-5p was upregulated in induced to osteogenic differentiation BMSCs. Furthermore, downregulation of MIAT dramatically promoted osteogenic differentiation, increased proliferation, and inhibited apoptosis in BMSCs; miR-150-5p inhibitor abrogated the effects of MIAT. In conclusion, lncRNA MIAT can regulate the proliferation, apoptosis, and osteogenic differentiation of BMSCs.

Hydroxy-Safflower Yellow A Alleviates Osteoporosis in Ovariectomized Rat Model by Inhibiting Carbonic Anhydrase 2 Activity.

Background: To investigate the therapeutic effect of Hydroxy-safflower yellow A (HSYA) on rat's osteoporosis and explore its potential mechanism of action. Methods: Bilateral ovariectomized female rats (OVX) were used to establish a postmenopausal rat model of osteoporosis. HSYA was given as an intervention, and estradiol was used as a positive control. The levels of serum alkaline phosphatase (ALP), calcium ion (Ca2+), and inorganic phosphorus (IP) were used to detect bone loss. Three months after modeling, the rats were sacrificed and the rat's ovaries, kidneys, tibia, and femur were used to calculate the organ index. The bone marrow of the femur of the rats was stained with Giemsa staining. The femur strength of rats was measured by INSTRON. The degree of osteoporosis was detected by pathological staining after decalcification of bone tissue. Predicted the main targets of HSYA in combination with bioinformatics, and the proteins related to osteoclast differentiation were detected in combination with western blotting. The effect of HSYA on the differentiation of RAW264.7 cells into osteoclasts was observed. Results: The Giemsa staining and serum test results showed that the operation was successful and affected bone metabolism. In the bone strength test, HSYA significantly increased the maximum threshold of femoral load in rats. Pathological examination showed that tibial cartilage, trabecular bone, and cortex significantly increased after treatment with HYSA. The number of osteoblasts increased while the number of osteoclasts decreased-elevated levels of type I and III collagen. Autodock was used for molecular docking of potential targets of HSYA. qPCR and western blot were used to show that the expression levels of CA2 and osteoclast differentiation-related proteins were significantly decreased after HSYA treatment. Cell level results showed that HSYA could inhibit the activity of osteoclasts and the ability of RAW264.7 cells to differentiate into osteoclasts. Conclusion: HSYA can inhibit the differentiation and formation of osteoclasts by inhibiting the expression of CA2 and relieving osteoporosis symptoms in OVX rats.

Knockdown of hsa_circ_0059955 Induces Apoptosis and Cell Cycle Arrest in Nucleus Pulposus Cells via Inhibiting Itchy E3 Ubiquitin Protein Ligase.

BACKGROUND: Circular RNAs (circRNAs) play an important role in the progression of intervertebral disc (IVD) degeneration (IVDD). Using bioinformatics analysis, we have found that the expression of circRNA hsa_circ_0059955 was significantly downregulated in IVDD tissues. However, the relevant mechanism of hsa_circ_0059955 in the progression of IVDD remains unclear. METHODS: CCK-8 and flow cytometry assays were used to evaluate cell proliferation and apoptosis. In addition, Western blot assay was used to detect the expressions of ITCH, p73, CDK2 in nucleus pulposus (NP) cells. Moreover, a puncture-induced IVDD rat model was established to explore the role of hsa_circ_0059955 in IVDD. RESULTS: The level of hsa_circ_0059955 was significantly decreased in IVDD tissues from IVDD patients. Itchy E3 ubiquitin protein ligase (ITCH) is the host gene of hsa_circ_0059955, and downregulation of hsa_circ_0059955 significantly decreased the expression of ITCH in NP cells. In addition, downregulation of hsa_circ_0059955 markedly inhibited proliferation and induced apoptosis and cell cycle arrest in NP cells. Moreover, in vivo study illustrated that overexpression of hsa_circ_0059955 ameliorated IVDD in rats. CONCLUSION: Downregulation of hsa_circ_0059955 could induce apoptosis and cell cycle arrest in NP cells in vitro, while overexpression of hsa_circ_0059955 attenuated the IVDD in a puncture-induced rat model in vivo. Therefore, hsa_circ_0059955 might serve as a therapeutic target for the treatment of IVDD.

Complete ureteral necrosis after injury sustained during lumbar disc surgery: A case report.

INTRODUCTION: Reports pertaining to ureteral injury sustained during lumbar disc surgery are rare; most ureteral injuries in this setting involve laceration or transection. PATIENT CONCERNS: We report a rare case of a 55-year-old man who presented with complete left ureteral necrosis 20 days after sustaining ureteral transection during lumbar disc surgery. DIAGNOSIS: The patient presented with seroperitoneum caused by left ureteral injury; post-operative histopathological examination of surgical specimen after discectomy had revealed ureter-like tissue. Exploratory laparoscopic surgery revealed necrosis of a long segment of ureter, which was not amenable to treatment with conventional methods. INTERVENTION: We used a spiral bladder muscle flap with vascular pedicles to repair the ureteral defect. OUTCOMES: Post-operative period was uneventful and the patient showed good recovery. CONCLUSION: Spiral bladder muscle flap with vascular pedicles may be used to repair extensive ureteric injury.

Long Non-Coding RNA ASB16-AS1 Functions as a miR-760 Sponge to Facilitate the Malignant Phenotype of Osteosarcoma by Increasing HDGF Expression.

PURPOSE: ASB16 antisense RNA 1 (ASB16-AS1) is a cancer-associated long non-coding RNA that contributes to tumorigenesis and tumor development. Nevertheless, to the best of our knowledge, whether and how ASB16-AS1 is implicated in osteosarcoma (OS) malignancy remains unclear and therefore warrants exploration. Our current study focused on making in-depth investigation of ASB16-AS1 in OS. In the present study, the expression pattern of ASB16-AS1 in OS tissues and cell lines was analyzed. In addition, we examined the clinical value of ASB16-AS1 for OS patients. Furthermore, we explored the impacts of ASB16-AS1 on the malignant phenotype of OS cells in vitro and in vivo as well as the underlying mechanism. METHODS: ASB16-AS1, microRNA-760 (miR-760) and hepatoma-derived growth factor (HDGF) expressions were measured using reverse transcription-quantitative PCR. Cell proliferation and apoptosis were evaluated using CCK-8 and flow cytometry analyses, respectively, and cell migration and invasion were determined via cell migration and invasion assays. RESULTS: ASB16-AS1 expression was significantly elevated in OS tissues and cell lines, and increased ASB16-AS1 expression was related to patients' tumor size, TNM stage, and distant metastasis. The overall survival rate of OS patients presenting high ASB16-AS1 expression was shorter than that of patients presenting low ASB16-AS1 expression. Reduced ASB16-AS1 expression inhibited OS cell proliferation, migration, and invasion; promoted cell apoptosis; and impaired tumor growth in vivo. Mechanistically, ASB16-AS1 served as a sponge for miR-760 and positively modulated the expression of its target HDGF. Finally, inhibiting miR-760 and restoring HDGF expression abolished the impacts of ASB16-AS1 knockdown on the malignant characteristics of OS cells. CONCLUSION: ASB16-AS1 is a novel oncogenic lncRNA in OS cells. ASB16-AS1 increased HDGF expression by sponging miR-760, thereby conferring cancer-promoting roles in OS. ASB16-AS1 is a potential early diagnostic and therapeutic target in OS.

miR-140-3p aggregates osteoporosis by targeting PTEN and activating PTEN/PI3K/AKT signaling pathway.

Osteoporosis (OP) is a systemic bone metabolic disorder, which negatively affects the quality of life in the elders and postmenopausal females. Healthy volunteers and postmenopausal females with OP were enrolled in the present study. Bone densitometry (BMD) was detected by dual-energy X-ray absorptiometry (DXA). CD14+PBMCs and C2C12 cells were cultured to induce osteoclast differentiation and osteoblast differentiation, respectively. The interaction between miR­140-3p and PTEN was predicted and verified by TargetScan 7.2 and dual luciferase reporter assay, respectively. miRNA/RNA level and protein level were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot, respectively. Cell proliferation and apoptosis were detected by 5-ethynyl-2'-deoxyuridine (EdU) staining and flow cytometry, respectively. Cell differentiation of CD14+PBMCs and C2C12 cells were detected by tartrate-resistant acid phosphatase (TRAP) staining and alizarin red staining, respectively. The activity of alkaline phosphatase (ALP) was detected by ALP assay. Differences were observed in age, body mass index (BMI), and BMD between the OP group and the control group. Higher miR­140-3p level and lower PTEN level were found in PBMCs of OP group compared to control group; there was a negative correlation between them in the serum of OP group. miR-140-3p targeted and downregulated the expression of PTEN. miR-140-3p inhibitor inhibited cell proliferation, differentiation, and promoted cell apoptosis of CD14+PBMCs; while promoted cell proliferation, differentiation and inhibited cell apoptosis of C2C12 cells, by targeting PTEN and inactivating PTEN/PI3K/AKT signaling pathway. These findings suggested a potential therapeutic role of miR-140-3p in the treatment of patients with OP.

[Assessment of correction effect in relation to fulcrum bending flexibility in adult idiopathic scoliosis patients].

OBJECTIVE: To study the relationship between the fulcrum bending flexibility (FBF) and correction rate (CR) of adult idiopathic scoliosis, and to explore the ability of FBF to assess the correction effect in relation to fulcrum bending flexibility. METHODS: 69 patients with adult idiopathic scoliosis with structural curves at thoracic or lumbarthoracic segments, 16 males and 53 females, aged 26.5 (19 - 53) were treated by pedicle screws instrumentation. Pre-operative standing and fulcrum bending films and postoperative standing X-ray film were taken. Cobb angle was measured. The data underwent regression analysis with the software SPSS 10.0. RESULTS: A regression formula was established: CR = 0.213 + 0.768 x FBF with P < 0.01. CONCLUSION: A definite linear relation exists between the FBF and CR of adult idiopathic scoliosis. By using the formula in proper samples, the effects of new instrumentation or correction technique can be objectively assessed.

[Association study of SIM2 gene polymorphisms with susceptibility to congenital scoliosis in a Chinese Han population].

OBJECTIVE: To investigate whether polymorphisms of SIM2 gene are associated with congenital scoliosis (CS) in a Chinese Han population. and explore the relationship of between polymorphisms of SIM2 and clinical phenotypes of CS. METHODS: A case-control design was employed in this study. A total of 127 patients (55 boys, 72 girls, mean age 12.90 y/o) diagnosed with CS admitted at Peking Union Medical College (PUMC) Hospital were enrolled between October 2005 and September 2007. The scoliosis-free control subjects (127 cases) at the same hospital during the same study period were frequency-matched to the cases on age (+/- 3 years) and gender. Genomic DNA was extracted by QIAamp DNA Blood Mini Kit from peripheral blood leukocytes of each subject who had signed informed consent. Based on genotype data from the International HapMap project, the main functional single nucleotide polymorphisms (SNPs) initially were selected. Case group were classified into different clinical phenotypes according to vertebral defect type, location of deformity, extent of developmental disruption, combined rib malformations and neural canal deformity. Genotying of all selected SNPs was done by SNPstream technology (Beckman Coulter SNPstream). All the data of SNPs with polymorphism were analyzed by association analysis based on a single SNP, the association analysis between phenotypes and SNPs. And pairwise linkage disequilibrium was calculated in the control population using Haploview 4.1 software. RESULTS: SNP1 (rs2073601), SNP2 (rs2073417) and SNP3 (rs2051397) of SIM2 are genotyped. SNP2 and SNP3 in linkage disequilibrium. No association (P > 0.05) is observed between SNP1, SNP2 and SNP3genotypes/allele polymorphisms and risk of CS and different clinical phenotypes. CONCLUSION: Genetic variants of SIM2 gene may not be associated with the susceptibility to CS and different clinical phenotypes of CS in Chinese Han population.

Catastrophic thoracolumbar spinal massive hematoma triggered by intraspinal anesthesia puncture: A CARE-compliant case report.

RATIONALE: Intraspinal anesthesia, the most common anesthesia type of orthopedic operation, is regarded as safe and simple. Despite of the rare incidence, puncture related complication of intraspinal anesthesia is catastrophic for spinal cord. Here we present an intradural hematoma case triggered by improper anesthesia puncture. The principal reason of this tragedy was rooted in the neglect of spine deformities diagnosis before anesthesia. To the best of our knowledge, there is no specific case report focusing on the intradural hematoma triggered by improper anesthesia puncture. PATIENT CONCERNS: Hereby a case of thoracolumbar spinal massive hematoma triggered by intraspinal anesthesia puncture was reported. The presenting complaint of the patient was little neurologic function improvement after surgery at 6-month follow-up. DIAGNOSES: Emergency MRI demonstrated that massive spindle-like intradural T2-weighted image hypointense signal masses from T12 to S2 badly compressed the dural sac ventrally, and his conus medullaris was at L3/4 intervertebral level with absence of L5 vertebral lamina. Hereby, the diagnoses were congenital spinal bifida, tethered cord syndrome, spine intradural hematoma, and paraplegia. INTERVENTIONS: Urgent surgical interventions including laminectomy, spinal canal exploration hematoma removal, and pedicle fixation were performed. The patient received both medication (mannitol, mecobalamin, and steroids) and rehabilitation (neuromuscular electric stimulation, hyperbaric oxygen). OUTCOMES: Postoperation, he had regained only hip and knee flexion at II grade strength. His neurologic function was unchanged until 3 weeks postoperation. Six-month follow-up showed just little neurologic function improvement, and the American Spinal Injury Association grade was C. LESSONS: By presenting an intradural hematoma case triggered by improper anesthesia puncture, we shared the treatment experience and discussed the potential mechanism of neurologic compromise. The principal reason for this tragedy is preanesthesia examination deficiency. Necessary radiology examinations must be performed to prevent misdiagnosis for spinal malformation.

[Association of PAX1 gene polymorphisms with susceptibility to congenital scoliosis in Chinese Han population].

OBJECTIVE: To investigate the association of polymorphisms of PAX1 gene with congenital scoliosis (CS) in Chinese Han population and the relationship between the PAX1 gene polymorphisms and the clinical phenotypes of CS. METHODS: Peripheral blood samples were collected from 127 CS patients, 55 male and 72 female, aged (12.9 +/- 4.3) (2 - 23), and 127 sex- and age-matched controls. Based on genotype data from the International HapMap project, the tagging single nucleotide polymorphisms (tSNPs) were selected using Haploview 4.0 software. Hardy-Weinberg equilibrium was analyzed both in the control and case groups. The case group was classified into different clinical phenotypes according to vertebral defect type, location of deformity, extent of developmental disruption, combined rib malformations, and neural canal deformity. Genotyping of all selected SNPs was done by SNPstream technology. The association between phenotypes and SNP was analyzed. Pairwise linkage disequilibrium was calculated in the control population using Haploview 4.0 software. RESULTS: The sites: SNP1 (rs17861031) and SNP2 (rs6047590), of PAX1 gene were genotyped and both polymorphisms were distributed in line with the Hardy-Weinberg equilibrium in these 2 groups. There was no linkage disequilibrium between these 2 SNPs. The genotype frequencies of SNP1AA, SNP1AG, SNP1GG, SNP2AA, SNP2AT, and SNP2TT of the case group were 2%, 26%, 72%, 2%, 19%, and 80% respectively, all not significantly different from those of the control group (2%, 26%, 72%, 2%, 26%, and 82% respectively, all P > 0.05). The allele frequencies of SNP1A, SNP1G, SNP2A, and SNP2T of the case group were 15%, 85%, 11%, and 89% respectively, all not significantly different from those of the control group (15%, 85%, 10%, and 90% respectively, all P > 0.05). No positive sites were found in different clinical phenotypes. CONCLUSION: The genetic variants of PAX1 gene may not be associated with the susceptibility to CS and different clinical phenotypes of CS in Chinese Han population.

MicroRNA-145-3p suppresses proliferation and promotes apotosis and autophagy of osteosarcoma cell by targeting HDAC4.

Studies have shown that miR-145-3p functions as a tumor suppressor and is associated with tumor growth and metastasis. This study intends to uncover the mechanism of a tumor suppressor of miR-145-3p. The expressions of miR-194 in osteosarcoma cell lines and tissues were monitored by real-time PCR. The proliferation ability was examined by MTT assay. The apoptosis and autophagy of cells were monitored by flow cytometry and microcopy, respectively. The regulation of miR-145-3p on HDAC4 was determined by luciferase assays and western blot assay. The results showed that miR-145-3p was significantly reduced in the osteosarcoma compared with the normal bone tissue. Overexpression of miR-145-3p significantly attenuated the proliferation and induced the apoptosis and autophagy of osteosarcoma cells. Furthermore, we demonstrated that miR-145-3p has inhibited the malignant behavior of osteosarcoma by down-regulating HDAC4 expression. These findings suggested that miR-145-3p may act as a tumor suppressor in osteosarcoma. MiR-145-3p/HDAC4 may be a novel therapeutic target in treatment of osteosarcoma.

Clinical effectiveness of treatment of combined upper thoracic spinal stenosis and multilevel cervical spinal stenosis with different posterior decompression surgeries.

OBJECTIVE: To explore the clinical effectiveness of various posterior decompression surgeries in the treatment of upper thoracic spinal stenosis combined with multilevel cervical spinal stenosis. METHODS: From January 2010 to December 2015, 22 consecutive patients with combined upper thoracic spinal stenosis and multilevel cervical spinal stenosis were treated with two different approaches of posterior decompression surgeries. In group A with 10 patients, both cervical and thoracic spinal decompression surgeries were performed simultaneously (one-stage surgery); in group B with 8 patients, cervical and thoracic spinal decompression surgeries were performed separately within three months (two-stage surgery). Based on Japanese Orthopedic Association (JOA) scores, improvement rate and extent of neurological function were calculated and the difference was compared between the two groups. RESULTS: There was no significant difference in demographic data between the two groups. However, compared with those of group B, both short-term and long-term improvement rate of neurological function in group A was higher (P < 0.05). In addition, the hospitalization cost was also lower in group A. CONCLUSION: Both one-stage and two-stage posterior decompression surgeries were effective in treating patient with upper thoracic spinal stenosis combined with multilevel cervical spinal stenosis; however, one-stage combined surgery was superior to two-stage surgery.

Thoracolumbar traumatic nucleus pulposus sequestration combined with a slight flexion distraction fracture: A rare case report and literature review.

RATIONALE: Traumatic nucleus pulposus sequestration (TNPS) usually occurs concurrently with severe destruction of bone. TNPS combined with a slight thoracolumbar flexion- distraction fracture, triggering a disastrous nerve injury, has rarely been reported. Due to the atypical radiologic manifestations, such a patient can easily be overlooked. PATIENT CONCERNS: Hereby, we present a TNPS patient as well as a slight thoracolumbar flexion-distraction fracture and serious neurologic symptoms. DIAGNOSES: T12 spinous process fracture, L1 flexion distraction fracture, thoracolumbar traumatic nucleus pulposuse sequestration and lower limbs incomplete paralysis INTERVENTIONS:: To avoid further neurologic compromise, an urgent laminectomy and exploration of the spinal canal was performed. OUTCOMES: After decompression OR and 4 months rehabilitation, the patient's neurologic function improved remarkably. LESSONS: A slight flexion-distraction fracture following injury is liable to eclipse the concurrence of TPNS. For this patient, a high-resolution MRI was needed to make a definitive diagnosis and guide surgery. Once TPNS has been diagnosed, sufficient decompression and discectomy surgery should be performed without delay.

[Assessment of curve flexibility in adolescent idiopathic scoliosis before operation].

OBJECTIVE: To compare the effects in assessing the curve flexibility of the adolescent idiopathic scoliosis (AIS) and predicting the outcomes of operation among different radiological techniques: supine lateral bending (SB), traction (Tr), and fulcrum bending radiographs. METHODS: 68 consecutive AIS patients, all with the single-curve types Ia/Ib/Ic according to the PUMC classification, divided into 4 groups according to the magnitude of Cobb's angle: moderate thoracic curve (n = 19, 40 degrees < Cobb's angle < or = 60 degrees ), severe thoracic curve (n = 13, Cobb's angle > 60 degrees ), moderate lumbar curve (n = 28, 35 degrees < Cobb's angle < or = 60 degrees ), and severe lumbar curve(n = 8, Cobb's angle > 60 degrees ) who were treated surgically underwent preoperative radiological evaluation including standing anteroposterior and lateral Tr, SB, and fulcrum bending radiographs. COBB angle was measured and the flexibility ratio was determined on each radiograph. The amounts of correction obtained by all radiographic methods were compared with the amount of surgical correction. RESULTS: The post-operative Cobb's angle of the moderate thoracic curve group was 9 degrees , not significantly different from that by fulcrum bending radiograph (P = 0.076), but significantly different from those by the other methods (both P < 0.01). The post-operative COBB angle of the severe thoracic curve group was 40 degrees , significantly different from all the radiographs before operation (all P < 0.01). The post-operative Cobb's angle of the moderate lumbar curve group was 4 degrees , significantly different from those by fulcrum bending and Tr radiographs (both P < 0.01) and that by SB (P = 0.013). The post-operative Cobb's angle of the severe lumbar curve group was 24 degrees , significantly different from those of anteroposterior and Tr radiograph (both P < 0.01) and those of fulcrum-bending and SB radiographs (P = 0.021 and P = 0.011). In the moderate thoracic curve group the operation correction rate was not significantly different from the flexibility rate by fulcrum-bending radiograph (P = 0.111), and was significantly different from the flexibility rates by SB and Tr radiographs (P = 0.011 and P = 0.000). In the severe thoracic curve group the operation correction rate was significantly different from the flexibility rates by different kinds of radiograph (all P = 0.111). In the moderate lumbar curve group the operation correction rate was significantly different from the flexibility rates by different kinds of radiograph (P < 0.111 or P = 0.019). In the severe lumbar curve group the operation correction rate was significantly different from the flexibility rates by different kinds of radiograph (P < 0.01 or P = 0.017). CONCLUSION: Fulcrum-bending radiography can better assess the flexibility and correction rate of thoracic curves in AIS, however, it can only predict those in moderate thoracic curves. Fulcrum-bending radiograph and SB radiograph are similar in predicting the flexibility in lumbar curves.

Syphilis of the lumbar spine: A case report and review of the literature.

RATIONALE: Tertiary syphilis can manifest as gummatous disease, but gumma of the spine has been extremely rarely reported. PATIENT CONCERNS: A 61-year-old male complained of worsening pain and numbness in both lower legs for four weeks. DIAGNOSES: Syphilis of the Lumbar Spine. INTERVENTIONS: Pedicle screw fixation (L3-S1) and posterior decompression of the vertebral canal at L4-5 were performed. OUTCOMES: The postoperative VAS score of both lower extremities decline to 2 from 7 at admission. Dorsal thumb extensor motor power (left/right) at day 7 postoperatively was 3/3 (versus admission: 1/1). Laboratory examinations showed normal white blood cell count (versus admission: 13.8 × 10/L; reference value: 4.00-10.00 × 10/L) and decline in C-reactive protein (20.35 mg/L versus admission: 77.43 mg/L; reference value: 0.00-10.00 mg/mL) and ESR (58 mm versus admission: 73 mm; reference value: 0-15 mm). LESSONS: Our case illustrates that although gummatous disease of the spine may be extremely rare, it should be considered in the differential diagnosis of tuberculosis or malignancy of the spine so as to avoid a wrong diagnosis and incorrect treatment.