Background Osteosarcoma is a highly aggressive primary bone malignancy in which distant metastasis critically determines patient prognosis. However, the molecular drivers of metastasis remain incompletely understood, making the elucidation of these drivers a research priority. Methods We integrated single-cell and bulk RNA sequencing data to identify cell populations associated with metastasis and to screen for candidate therapeutic targets. Bioinformatics tools predicted upstream transcriptional regulators. Functional roles and transcriptional regulation were assessed via wound-healing, Transwell, and luciferase reporter assays. Results Single-cell analysis of osteosarcoma heterogeneity identified the osteoblastic subcluster C2_OB as a major contributor to metastasis. cNMF analysis defined a metastasis-associated expression program (module 2) highly active in C2_OB, within which S100A13 emerged as a novel signature gene. S100A13 was overexpressed in osteosarcoma and correlated with metastasis and poor prognosis. Functional assays demonstrated that S100A13 promoted cell migration and invasion. Mechanistically, the transcription factor SP1 directly binds to the S100A13 promoter, activates its transcription, and thereby enhances the metastatic capacity of osteosarcoma cells. Conclusion This study identifies S100A13 as a biomarker for osteosarcoma metastasis and poor prognosis, and delineates an SP1/S100A13 axis that drives metastatic progression.

Background Osteonecrosis of the femoral head (ONFH) is a refractory orthopedic disease with a high disability rate. Long-term administration of steroids is the most common pathogenic factor for non-traumatic ONFH. Early diagnosis of steroid-induced osteonecrosis of the femoral head (SONFH) is difficult and mainly depends on imaging.Objectives The objectives of this review were to examine the pathological mechanisms of SONFH, summarize related markers of SONFH, and identify areas for future studies.Methods We reviewed studies on pathological mechanisms and related markers of SONFH and discussed the relationship between them, as well as clinical applications and the outlook of potential markers.Results The pathological mechanisms of SONFH included decreased osteogenesis, lipid accumulation, increased intraosseous pressure, and microcirculation disruption. Differential proteomics and genomics play crucial roles in the occurrence, progression, and outcome of SONFH, providing novel insights into SONFH. Additionally, the biological functions of mesenchymal stem cells (MSCs) and exosomes (Exos) in SONFH have attracted increasing attention.Conclusions The pathological mechanisms of SONFH are complex. The related markers mentioned in the current review can predict the occurrence and progression of SONFH, which will help provide effective early clinical prevention and treatment strategies for SONFH.

Traumatic spinal cord injury is potentially catastrophic and can lead to permanent disability or even death. China has the largest population of patients with traumatic spinal cord injury. Previous studies of traumatic spinal cord injury in China have mostly been regional in scope; national-level studies have been rare. To the best of our knowledge, no national-level study of treatment status and economic burden has been performed. This retrospective study aimed to examine the epidemiological and clinical features, treatment status, and economic burden of traumatic spinal cord injury in China at the national level. We included 13,465 traumatic spinal cord injury patients who were injured between January 2013 and December 2018 and treated in 30 hospitals in 11 provinces/municipalities representing all geographical divisions of China. Patient epidemiological and clinical features, treatment status, and total and daily costs were recorded. Trends in the percentage of traumatic spinal cord injuries among all hospitalized patients and among patients hospitalized in the orthopedic department and cost of care were assessed by annual percentage change using the Joinpoint Regression Program. The percentage of traumatic spinal cord injuries among all hospitalized patients and among patients hospitalized in the orthopedic department did not significantly change overall (annual percentage change, -0.5% and 2.1%, respectively). A total of 10,053 (74.7%) patients underwent surgery. Only 2.8% of patients who underwent surgery did so within 24 hours of injury. A total of 2005 (14.9%) patients were treated with high-dose (≥ 500 mg) methylprednisolone sodium succinate/methylprednisolone (MPSS/MP); 615 (4.6%) received it within 8 hours. The total cost for acute traumatic spinal cord injury decreased over the study period (-4.7%), while daily cost did not significantly change (1.0% increase). Our findings indicate that public health initiatives should aim at improving hospitals' ability to complete early surgery within 24 hours, which is associated with improved sensorimotor recovery, increasing the awareness rate of clinical guidelines related to high-dose MPSS/MP to reduce the use of the treatment with insufficient evidence.

Objective:To observe the long-term effects of percutaneous vertebroplasty (PVP) and percutaneous kyphoplasty (PKP) on osteoporotic vertebral compressive fractures (OVCFs).Methods:The clinical data of 680 OVCFs patients admitted to the First Affiliated Hospital of Zhengzhou University from June 2005 to December 2015 were retrospectively analyzed. Patients who died during follow-up (more than 5 years) were excluded, and 390 patients were finally included in the study. According to the treatment methods, the patients were divided into PVP group (210 cases) and PKP group (180 cases). The operation time, bone cement injection amount and surgical complications were compared between the two groups. The visual analogue scale (VAS) score and imaging changes of the injured vertebrae were compared between the two groups before operation, 3 days after operation, and at the last follow-up.Results:The operation time of the PVP group was (33.1±10.0)min, which was shorter than the (39.1±12.8)min of the PKP group, while the amount of bone cement injection of the PVP group was (3.5±1.1)ml, less than the (4.8±1.2)ml of the PKP group, and the differences were statistically significant ( t=-5.592, -12.779; all P<0.01). There was no significant difference in the leakage rate of bone cement and incidence of adjacent vertebral fractures between the two groups ( P>0.05). Compared with the preoperative scores, the VAS scores 3 day after the operation and at the last follow-up of the two groups were lower, and the differences were statistically significant (all P<0.05), but there was no significant difference between the two groups ( P>0.05). The postoperative height of the vertebral body in the PVP group and the PKP group increased by (3.9±2.3)mm and (5.6±2.6)mm, respectively, and the Cobb angle of the injured vertebrae decreased by (4.2±2.3)° and (6.4±2.8)°, respectively, and the differences were statistically significant ( t=-7.069, -7.930; all P<0.01); while there was no significant difference between the two groups in the loss of vertebral body height and Cobb angle at the last follow-up (all P>0.05). Conclusions:PVP and PKP can significantly reduce the pain symptoms of patients with OVCFs. PKP is better than PVP in the recovery of vertebral height and the improvement of Cobb angle. Both PVP and PKP have long-term effects and safety in the treatment of OVCFs.

Purpose: The primary purpose of this systematic review and meta-analysis was to investigate the impact of prior arthroscopy on postoperative revisions, complications, and other clinical outcomes after conversion total lower extremity arthroplasty. Methods: Two individual researchers conducted the platform searches on the Embase, PubMed, Cochrane Central, and Google Scholar electronic databases from inception to June 02, 2021. We identified cohort trials that compared the outcomes of patients who underwent primary THA or TKA in the prior arthroscopy or control groups. The primary outcome was revision, and secondary outcomes included reoperation, patient-reported outcomes, and postoperative complications. A modified version of the Downs and Black tool was used to assess the methodological quality of the non-randomized cohort studies. Results: Of the 23 included studies with 319946 cases, 18 were matched retrospectively and five were non-matched retrospectively. Methodological quality was high in ten studies and moderate in thirteen studies. Our analysis demonstrated that TKA or THA patients with prior arthroscopy were associated with an increased risk of revision, reoperation, infection, and aseptic loosening. THA patients with prior arthroscopy were also associated with an increased risk of dislocation. Furthermore, there were no significant intergroup differences in periprosthetic fracture, range of motion, Harris Hip Score, or Knee Society Score. Conclusion: Arthroscopy performed before total lower extremity arthroplasty substantially increased the revision, reoperation, infection, and aseptic loosening rates. THA patients with prior arthroscopy were also associated with an increased risk of dislocation. Patients should be counseled on the potential increased risks associated with conversion total lower extremity arthroplasty after prior arthroscopy. Further research is needed to better characterize these findings.

Abstract Background Circular RNA was reported tightly associated with the incidence and progress of tumor, including osteosarcoma (OS). Circ_0000530, a newly discovered circular RNA, the expression and function were not investigated in OS until now. Here, we aimed to explore it. Materials and Methods By adopting real-time quantitative polymerase chain reaction (qRT-PCR), the level of circ_0000530, miR-198, and cyclin-dependent kinase 6 (CDK6) was identified. Moreover, flow cytometry and transwell assays were adopted for investigating cell cycle distribution, cell migration as well as invasion abilities. Furthermore, using dual-luciferase reporter gene experiments, the correlation between miR-198 and circ_0000530 or CDK6 was revealed. CDK6 protein levels were detected by western blotting (WB). In addition, the impact of circ_0000530 silencing on the growth of OS tumor in vivo was also validated. Results Circ_0000530 levels were raised in OS tissues and cells. Besides, it was shown that knockdown of circ_0000530 could hinder the proliferation, migration as well as invasion of OS cells, also inducing cell cycle arrest. Circ_0000530 could be adsorbed to miR-198, and miR-198 inhibitor was capable of reversing the inhibitory impact of silenced circ_0000530 on OS progression. Silencing of circ_0000530 could hinder CDK6 expression whereas miR-198 inhibitor could be used to recover the impact. Apart from that, circ_0000530 knockdown could reduce OS tumor growth through the regulation of CDK6 expression based on miR-198 in vivo. Conclusion Down-regulation of circ-0000530 inhibits the development of osteosarcoma through the regulation of miR-198 and CDK6, providing new ideas for diagnosing and treating osteosarcoma.

Objective:To investigate the effect of nucleosome assembly protein 1 like protein 1 (NAP1L1) on the proliferation of nasopharyngeal carcinoma cells and its molecular mechanism.Methods:A total of 60 cases of nasopharyngeal carcinoma tissues and adjacent tissues collected in our hospital from January 2017 to January 2021 were selected as the research objects. The expression levels of NAP1L1 protein in adjacent tissues and nasopharyngeal carcinoma tissues were analyzed by immunohistochemistry. The low differentiated nasopharyngeal carcinoma cell line SUNE-1 was infected with NAP1L1 short hairpin RNA (shRNA) lentivirus. The cell lines of control group and NAP1L1 KD group were established. The effects of NAP1L1 on the proliferation of nasopharyngeal carcinoma cells were analyzed by cell counting kit-8 (CCK-8), clone formation test and in vitro tumor transplantation test. The cell cycle levels of the two groups were analyzed by flow cytometry. The protein expression levels of cyclin dependent kinase 4 (CDK4), cyclin dependent kinase 6 (CDK6) and Cyclin D1 (CCND1) in tumor tissues and adjacent tissues were analyzed by Western blotting. The measurement data between groups were compared by t-test. Results:NAP1L1 protein was mainly expressed in the nucleus. The quantitative results showed that the expression intensity of NAP1L1 protein in adjacent tissues (31.64±7.01) was significantly lower than that in nasopharyngeal carcinoma (91.80±15.18, t=27.87, P<0.05). The 48 h and 72 h absorbance values ( A) of cells in the control group (1.08±0.08, 1.89±0.11) were significantly higher than those in the NAP1L1 KD group (0.79±0.08, 1.31±0.05, t=6.328, 11.570, P<0.05). The number of clonogenic cells in the control group (109.50±14.45) was significantly greater than that in the NAP1L1 KD group (66.33±7.69, t=6.462, P<0.05). At 30th day after inoculation, the tumorigenic volume and weight of cells in the control group [(1 072.90±114.87) mm 3, (4.96±0.52) g] were significantly increased as compared with those in the NAP1L1 KD group [(855.90±55.54) mm 3, (3.27±0.23) g, t=5.387, 9.425, P<0.05]. The G 0/G 1 phase ratio of cells in the control group [(55.03±3.09)%] was significantly lower than that in the NAP1L1 KD Group [(74.88±2.16)%, t=12.890, P<0.05]. The G 2/M phase ratio of cells in the control group [(5.61±0.63)%] was significantly lower than that in the NAP1L1 KD group [(14.01±1.03)%, t=17.080, P<0.05]. The S phase ratio of cells in the control group [(30.27±1.79)%] was significantly higher than that in the NAP1L1 KD group [(11.44±2.34)%], and the difference was statistically significant ( t=15.650, P<0.05). The expression levels of CDK4, CDK6 and CCND1 proteins in control group (1.23±0.06, 1.03±0.09, 0.88±0.05) were significantly higher than those in NAP1L1 KD group (0.41±0.07, 0.60±0.05, 0.30±0.05, t=21.330, 10.560, 19.980, P<0.05). Conclusion:NAP1L1 is highly expressed in nasopharyngeal carcinoma. It can regulate the changes of cell cycle by regulating the expression level of tumor cell cycle related proteins, and then affect the ability of malignant proliferation of tumor cells.

Objective:To observe the effect of miR-23a-3p on osteogenesis differentiation of bone marrow mesenchymal stem cells (BMSCs) by regulating Runx2 gene.Methods:The expression level of miR-23a-3p in serum of patients with postmenopausal osteoporosis (PMOP) (30 cases) and normal female volunteers (30 cases) was detected. After the establishment of female mouse osteoporosis model, BMSCs were extracted, and miR-23a-3p inhibitor and miR-NC were transfected into BMSCs respectively to detect the expression level of miR-23a-3p. The expression levels of Runx2, Osteocalcin (OCN) and Collagen Ⅰ mRNA were detected, and the contents of alkaline phosphatase (ALP) in cells were detected by enzyme linked immunosorbent assay (ELISA) kit. HEK293 cells were cultured, and miR-23a-3p mimics and miR-NC were co-transfected into wild-type Runx2 3′untranslated regions (3′UTR) (WT) or mutant Runx2 3′UTR (Mut) recombinant plasmids, respectively. The lucifase activity was detected using dual-luciferase reporter systerm. Finally, miR-23a-3p inhibitor and miR-23a-3p inhibitor+ siRunx2 were transfected into BMSCs respectively, and the expression level of Runx2 protein, OCN, Collagen Ⅰ and ALP contents were detected. The t test was used to compare the two samples between the two groups. Results:The expression level of miR-23a-3p in serum of PMOP patients was significantly higher than that of control group (0.872±0.307 vs. 0.382±0.183, t=7.514, P<0.05). The expression level of miR-23a-3p in BMSCs transfected with miR-23a-3p inhibitor was significantly lower than that in the Mir-NC group (0.480±0.045 vs. 1.361±0.113, t=-16.213, P<0.05). The expression levels of Runx2 protein, OCN, Collagen Ⅰ mRNA and ALP content in BMSCs transfected with miR-23a-3p inhibitor were increased as compared with those in BMSCs with mir-NC transfection [0.608±0.065 vs. 0.234±0.041, 0.523±0.053 vs. 0.229±0.046, 0.652±0.060 vs. 0.381±0.030, (5.569±0.377) U/L vs. (2.513±0.308) U/L, t=9.409, 9.036, 10.846, 14.036, P<0.05]. Bioinformatics analysis and dual-luciferase reporter gene assay further confirmed that Runx2 was a direct target of miR-23a-3p. The expression levels of Runx2 protein, OCN, Collagen Ⅰ mRNA and ALP contents in BMSCs transfected with miR-23a-3p inhibitor+ siRunx2 were significantly lower than those in BMSCs transfected with miR-23a-3p inhibitor [0.373±0.051 vs. 0.600±0.083, 0.321±0.015 vs. 0.645±0.030, 0.417±0.027 vs. 0.617±0.065, (3.733±0.056) U/L vs. (6.751±0.091) U/L, t=5.188, 21.491, 6.414, 63.040, P<0.05]. Conclusion:miR-23a-3p inhibits the osteogenic differentiation of BMSCs by negatively regulating the expression of Runx2 gene.

Objective: Osteosarcoma is the most common malignancy in the skeletal system; studies showed an important role of miRNAs in tumorigenesis, indicating miRNAs as possible therapeutic molecules. This study found abnormal hsa-miR-557 expression levels in osteosarcoma and tried to explore the potential function and the mechanism.Methods: Differential expression genes of osteosarcoma were analyzed using GSE28423 from the GEO database. Survival analysis of miRNAs was conducted with data obtained from the TARGET-OS database. STRING and miRDIP were used to predict target genes of hsa-miR-557; KRAS was then verified using dual-luciferase reporter assay. Expression of genes was detected by qPCR, and levels of proteins were detected by Western blot. The proliferation ability of cells was detected by CCK-8 and cell cycle analysis. Tumor formation assay in nude mice was used to detect the influence of osteosarcoma by hsa-miR-557 in vivo.Results: Analysis from the GEO and TARGET databases found 12 miRNAs that are significantly related to the osteosarcoma prognosis, 7 downregulated (hsa-miR-140-3p, hsa-miR-564, hsa-miR-765, hsa-miR-1224-5p, hsa-miR-95, hsa-miR-940, and hsa-miR-557) and 5 upregulated (hsa-miR-362-3p, hsa-miR-149, hsa-miR-96, hsa-miR-744, and hsa-miR-769-5p). CCK-8 analysis and cell cycle analysis found that hsa-miR-557 could significantly inhibit the proliferation of osteosarcoma cells. The tumor formation assay in nude mice showed that tumor sizes and weights were inhibited by hsa-miR-557 transfection. Further studies also proved that hsa-miR-557 could target the 3 ' UTR of KRAS and modulate phosphorylation of downstream proteins.Conclusion: This study showed that hsa-miR-557 could inhibit osteosarcoma growth both in vivo and in vitro, by modulating KRAS expression.

Editor's Note: this Article has been retracted; the Retraction Note is available at https://doi.org/10.1038/s41598-021-83145-9.

目的 观察circSAFB2通过微小RNA(miR)-1301-3p/zeste基因增强子同源物2(EZH2)对骨肉瘤细胞凋亡和侵袭的影响.方法 选用骨肉瘤细胞株MG63,实时定量反转录聚合酶链反应(RT-qPCR)检测MG63细胞和正常成骨细胞中circSAFB2、miR-1301-3p和EZH2 mRNA的表达.将circSAFB2小干扰RNA(siRNA)慢病毒感染骨肉瘤细胞,用流式细胞术检测细胞凋亡率,Transwell检测细胞侵袭能力.培养HEK293细胞,将miR-1301-3p mimic和miR-NC分别与野生型circSAFB2(Wt)或突变型circSAFB2(Mut)重组报告基因质粒共转染,将miR-1301-3p mimic和miR-NC分别与野生型EZH2 3'端非编码区(3'UTR)(Wt)或突变型EZH2 3'UTR(Mut)重组报告基因质粒共转染,使用双荧光素酶报告系统检测荧光素酶活性.蛋白质印迹法(Western blot)检测细胞中上调miR-1301-3p表达后EZH2表达水平的变化.最后分别转染circSAFB2 siRNA、miR-1301-3p mimic、共转染 circSAFB2 siRNA 和 pcDNA3.1-EZH2、共转染 miR-1301-3p mimic 和 pcDNA3.1-EZH2,通过流式细胞术、Transwel1分别检测各组细胞凋亡和侵袭能力.组间比较采用t检验.结果 RT-qPCR结果显示,骨肉瘤细胞MG63中circSAFB2、EZH2 mRNA表达量高于正常成骨细胞(2.263±0.154 比 1.000±0.093、4.155±0.406 比 1.000±0.092,t=15.686、16.955,P＜0.05);miR-1301-3p表达量低于正常成骨细胞(0.333±0.031 比 1.000±0.120,t=12.095,P＜0.05).感染 circSAFB2 siRNA骨肉瘤细胞的凋亡率高于感染circSAFB-NC组和Blank组[(22.48±2.27)％比％(5.92±0.57)％、(5.61±0.62)％,F=142.710,P＜0.05],侵袭数量低于感染 circSAFB-NC 组和 Blank 组[(49.333±7.024)比(112.667±14.978)、(110.667±14.048),F=24.767,P＜0.05].生物信息学分析及双荧光素酶报告基因检测进一步证实了 CircSAFB2靶向miR-1301-3p,同时EZH2是miR-1301-3p的靶基因.circSAFB2 siRNA和pcDNA3.1-EZH2共转染的凋亡率低于单独转染circSAFB2 siRNA 组[(12.07±0.92)％比(19.58±2.13)％,F=51.211,P＜0.05],侵袭数量高于单独转染circSAFB2siRNA组[(77.333±12.741)比(47.667±7.506),F=13.595,P＜0.05].miR-1301-3p mimic 和 pcDNA3.1-EZH2 共转染的凋亡率低于单独转染 miR-1301-3p mimic 组[(14.07±1.42)％比(20.73±1.99)％,F=51.211,P＜0.05],侵袭数量高于单独转染 miR-1301-3p mimic 组[(83.667±12.741)比(50.667±8.622),F=13.595,P＜0.05].结论 circSAFB2 通过 miR-1301-3p/EZH2调控骨肉瘤细胞的凋亡和侵袭的作用.

目的 探讨伴轻微颈椎外伤的脊髓型颈椎病(CSM)患者脊髓信号强度和病程等临床特点与骨质疏松(OP)的相关性.方法 纳入2015年8月至2021年12月郑州大学第一附属医院收治的72例伴轻微颈椎外伤的CSM患者.患者在入院后均接受颈椎核磁共振成像(MRI)检查,依据Yukawa分级法进行颈髓信号强度分级,使用改良日本整形外科协会(JOA)评分评价患者的神经功能障碍程度.通过电化学发光法检测Ⅰ型胶原羧基端肽β特殊序列(β-CTX)、Ⅰ型前胶原氨基端前肽(P1NP)和骨钙素(OC),通过双能X线骨密度仪测量骨密度(BMD).采用R统计分析法评估CSM患者脊髓信号强度和病程与OP的相关性.结果 Yukawa分级越高,3-CTX升高越明显;Yukawa 0级和1级患者P1NP低于正常值,而Yukawa 2级患者P1NP仍在正常范围内;各组患者OC值为(17.31±9.76)ng/ml,与 Yukawa 分级无相关(t=-0.32,P＞0.05).CSM 病程≥24 个月患者髋部和腰椎Z值较CSM病程＜24个月患者更低,差异有统计学意义(髋部:W=1 100.5,P＜0.01;腰椎:W=1 090.0,P＜0.01);CSM病程≥24个月患者与CSM病程＜24个月患者比较,髋部较腰椎Z值变化更明显.结论 CSM患者相对于健康人群BMD更低,且髋部较腰椎BMD下降更加明显,CSM患者颈椎在受到轻微外伤后容易导致脊髓再损伤,β-CTX可在短期内升高,需重视此类患者OP的预防和治疗.

RNA N6-methyladenosine (m6A) level is closely associated with neurodevelopment and central nervous system dysfunctions including spinal cord injury (SCI). M6A level can be dynamically regulated by m6A methyltransferases and demethylases. In this text, the roles of m6A demethylase FTO alpha-ketoglutarate dependent dioxygenase (FTO) in SCI development along with its m6A-dependent regulatory mechanisms were investigated in hypoxia-induced PC12 cell injury model. The results showed that FTO was low expressed in spinal cord tissues of rats after contusive SCI and hypoxia-treated PC12 cells. FTO knockdown alleviated hypoxia-induced PC12 cell injury. FTO loss increased GADD45B expression and m6A level in PC12 cells. GADD45B knockdown weakened the protective effects of FTO depletion on hypoxia-treated PC12 cells. FTO regulated GADD45B expression in an IGF2BP2-dependent manner. In conclusion, FTO knockdown mitigated the injury of hypoxia-induced PC12 cells by up-regulating GADD45B in an IGF2BP2-dependent manner.

Objective:To explore the safety of perioperative intravenous tranexamic acid in male smoking patients.Methods:According to the inclusion and exclusion criteria, a total of 120 patients who underwent knee arthroplasty in our hospital from January 2017 to December 2020 were screened and divided into three groups: intravenous tranexamic acid was used in smoking patients; tranexamic acid was not used in smoking patients and tranexamic acid was used in non-smokers. Inclusion criteria were as follows: patients with moderate to severe osteoarthritis, men aged 60-80 years; initial knee replacement; patients and their family members agreed to participate in the trial and sign an informed consent form; the experimental group had a history of smoking more than 1 year before surgery and did not quit smoking; the control group had no history of smoking or quit smoking for more than 1 year. Exclusion criteria were as follows: simultaneous knee arthroplasty on both sides; no serious complications before the operation; no coagulation disease before the operation; no history of the use of coagulation drugs before the operation; no tranexamic acid allergy; no knee joint infection. All patients underwent knee arthroplasty by the same chief physician. Patients were given 1.0 g tranexamic acid intravenously before skin incision and after implantation of the prosthesis. Statistics of patients′ operation time, intraoperative blood loss, postoperative drainage, hidden blood loss, blood transfusion, preoperative and postoperative coagulation indicators, postoperative venous thrombosis events and complications. The chi-square test and independent sample t-test were used for data analysis. Results:The postoperative Hct levels of smokers and non-smokers after tranexamic acid were significantly higher than those in the non-tranexamic acid group (smoking+ tranexamic acid: 0.36±0.05; smoking only group: 0.30±0.08; pure tranexamic acid group: 0.35±0.10, t1=1.819, P1<0.01, t3=2.469, P3<0.05), and postoperative Hct changes (smoking+ tranexamic acid: 0.12±0.02; smoking only group: 0.18±0.05; pure tranexamic acid group: 0.10±0.07, P1<0.01, P2<0.01) and Hb change values (smoking+ tranexamic acid: 45.50±12.20; smoking only group: 55.21±8.48; pure tranexamic acid group: 41.50±12.20, t1=7.047, P1<0.01, t3=5.882, P3<0.01). Postoperative drainage (661.80±358.11, t1=2.319, P1<0.05, t3=2.914, P3<0.01), hidden blood loss [(451.51±389.73) ml, t1=2.782, P1<0.01, t3=3.947, P3<0.01] and calculated total blood loss [(1 113.31±729.33) ml, t1=2.678, P1<0.01, t3=3.518, P3>0.05] were significantly higher than in smoking+ tranexamic acid group [postoperative drainage volume (500.82±254.11) ml, hidden blood loss (261.22±187.73) ml, total blood loss (752.03±442.82) ml] and the tranexamic acid group [postoperative drainage volume (500.82±254.11) ml, hidden blood loss (197.21±119.14) ml, total blood loss (660.35±362.48) ml]. The probability of postoperative blood transfusion was also significantly increased (smoking+ tranexamic acid group: 7/33; 18/22 in the smoking group; 8/32 in the tranexamic acid group, χ12=0.704, P1<0.01, χ32=5.698, P3<0.05). Conclusion:The use of tranexamic acid in the perioperative period of joint replacement surgery in smoking patients can significantly reduce the amount of bleeding and the chance of blood transfusion, but does not significantly increase the incidence of deep vein thrombosis in patients.

Ginsenoside Rb1 (GRb1), a major ingredient of ginseng, has been found to be a potential protective agent in spinal cord injury (SCI) and in activated microglia-induced neuronal injury. This study discovered that GRb1 could facilitate miR-130b-5p expression in SCI rats and Toll-like receptor 4 (TLR4; a crucial player in inflammation) was a potential target of miR-130b-5p. Hence, we further investigated whether GRb1 could relieve SCI by reducing microglia-mediated inflammatory responses and neuronal injury via miR-130b-5p/TLR4 pathways. The results showed that GRb1 alleviated SCI through inhibiting neuronal apoptosis and proinflammatory factor expression via increasing miR-130b-5p.GRb1 weakened the damage of activated microglia to neurons through upregulating miR-130b-5p. miR-130b-5p attenuated activated microglia-induced neuron injury via targeting TLR4. GRb1 inactivated TLR4/nuclear factor-kappa B (NF-kappa B) activation and inhibited proinflammatory cytokine secretion by increasing miR-130b-5p in activated microglia. As a conclusion, GRb1 alleviated SCI through reducing activated microglia-induced neuronal injury via miR-130b-5p/TLR4/NF-kappa B axis, providing a deep insight into the molecular basis of GRb1 in the treatment of SCI.

Objective:To explore the regulation of microRNA (miRNA, miR)-345-5p on osteoblast.Methods:MiRNA expression data sets were analyzed to find differentially expressed miRNAs. The pLVX-IRES-ZsGreen1 plasmid was used to construct overexpression vectors. The proliferation ability of the three groups of cells were tested by cell counting kit-8 (CCK-8) and growth, the dual luciferase assay verified the interaction of miR-345-5p and Smad1 gene. Cellular studies were established to verify the interaction between miR-345-5p and Smad1 genes in rat osteoblasts. Student- t test and ANOVA were used for analysis. Results:The bio-information analysis showed miR-345-5p and its target molecular Smad1. After the miR-345-5p plasmid was transformed into rat osteoblasts, CCK-8 showed miR-345-5p plasmid significantly enhanced the activity of osteoblasts (48 h: q=7.453, P<0.01, 72 h: q=15.34, P<0.01). Growth assay showed the number of osteoblasts with miR-345-5p plasmid was significantly lower than that of the control group at 48 h ( q=3.814, P<0.05), miR-345-5p mutation ( P<0.01) and the control group ( P<0.01) at 72 h. The dual-luciferase assay showed miR-345-5p reduced the fluorescence activity of wild-type Smad1-3′untranslated regions (3′UTR) plasmid. MiR-345-5p significantly reduced the activity of rat osteoblasts through modulating Smad1 in cellular experiment. Conclusion:MiR-345-5p regulates osteoblasts through Smad1, thus participating in fracture healing.

Objective:To investigate the difference of curative effect between zero-profile bridge-shaped locking cage (ROI-C) and anterior cage combined with titanium plate fixation in the treatment of two-level and three-level cervical spondylotic myelopathy.Methods:A total of 85 patients (43 males and 42 females), aged 52.3±8.0 years (range from 28 to 66 years) with bi- and three-level cervical spondylotic myelopathy who received surgical treatment from June 2017 to October 2019 were retrospectively analyzed. There were 63 cases of two levels and 22 cases of three levels. 45 cases were treated with zero-profile bridge-shaped locking cage ROI-C (ROI-C group), and 40 cases with anterior cage combined with titanium plate fixation (titanium plate group). The main observation indicators include operation time, intraoperative blood loss, cervical Cobb angle, fusion segment Cobb angle, average intervertebral height, pain visual analogue scale (VAS), Japanese Orthopaedic Association (JOA) Score and neck disability index (NDI).Results:All of 85 patients were followed up for 16.9±2.0 months (range 12 to 22 months). The operation time of two-level ROI-C group was 110.37±8.25 min, which was shorter than 139.5±10.54 min of titanium plate group; the intraoperative blood loss was 15.74±8.10 ml, which was less than 23.71±9.70 ml of titanium plate group; the operation time of three-level ROI-C group was 130.00±5.70 min, which was shorter than 162.83±5.59 min of titanium plate group, while the difference in the intraoperative blood loss between the two groups had no statistical significance. One year after operation, Cobb angle of cervical vertebra in double and three-level ROI-C groups were 15.31°±1.55° and 15.20°±0.42°, respectively, which were largerthan 11.23°±2.03° and 9.20°±1.14° before operation; in titanium plate group, they were 15.89°±1.13° and 16.08°±1.88°, which were higher than 11.25°±2.01° and 9.00°±1.60° before operation, and the differences had statistical significance. The differences between the two groups before operation and 1 year after operation had no statistical significance. One year after operation, the VAS scores of double and three-level ROI-C groups were 1.83±0.66 points and 2.60±0.52 points, respectively, which were less than the preoperative 7.49±0.51 points and 7.60±0.52 points; the titanium plate group was 1.79±0.50 points and 2.41±0.51 points, which were less than the preoperative 7.61±0.63 points and 7.42±0.52 points, and the differences had statistical significance. There was no significant difference between the two groups before operation and 1 year after operation. One year after operation, the JOA scores of double and three-level ROI-C groups were 15.00±0.84 points and 14.70±0.95 points, respectively, which were higher than the preoperative 7.20±0.87 points and 6.60±1.27 points; the scores of titanium plate group were 15.29±0.85 points and 14.83±0.58 points, which were higher than the preoperative 6.89±1.03 points and 6.92±0.67 points, and the differences had statistical significance. The differences between the two groups had no statistical significance. The postoperative JOA improvement rate was excellent. Postoperative dysphagia occurred in 1 case (2.22%, 1/45) in ROI-C group and 8 cases (20.00%, 8/40) in titanium plate group, and the difference in the incidence rate between two groups had statistical significance ( χ2=5.32, P=0.02). Conclusion:Both ROI-C and anterior cage combined with titanium plate fixation in the treatment of double and three-level cervical spondylotic myelopathy can achieve good short-term clinical efficacy, with shorter operation time and lower incidence rate of postoperative dysphagia using ROI-C.

Objective:To explore the effect of long non-coding RNA (lncRNA)-TCONS_00041960 on adipogenic and osteogenic genes expression in rat model of steroid-induced osteonecrosis of the femoral head.Methods:Totally, 64 healthy SD rats were randomly divided into 4 groups: (1) normal control group (blank group) (2 ml physiological saline was injected into gluteal muscle for 3 days). (2) hormone group (Dex group) [Dexamethasone (20 mg/kg) was injected into gluteal muscle for 3 days to establish steroid-induced femoral head necrosis model]. (3) unrelated sequence group (Ex-NC+ Dex group) [Dexamethasone was given in the same way, and bone marrow mesenchymal stem cells (BMSCs) transfected with lentivirus of unrelated sequence were injected into one side of femoral head]. (4) Dexamethasone+ recombinant lentivirus group (Ex-Lnc+ Dex group) [Dexamethasone was given in the same way, and BMSCs transfected with recombinant lentivirus vector lncRNA-TCONS_00041960 were injected into one side of femoral head]. At 4th and 8th week, total RNA and total protein were extracted from the femoral head tissue of each group. The expression of the peroxisome proliferator-activated receptor-γ (PPARγ), runt-related transcription factor 2 (Runx2) mRNA and protein was detected by TaqMan real-time quantitative polymerase chain reaction (Real-time PCR) and Western blotting at 4th week and 8th week of the experiment. The comparison between the two groups was performed by t test; the numerical variable data statistics were performed by analysis of variance, and the comparison between groups was performed by the least significant difference method (LSD). Results:At 4 weeks, the expression levels of PPARγ mRNA and protein in the femoral head cells in the Ex-Lnc+ Dex group (1.129±0.024 and 0.366±0.007) were not significantly different from those in the blank group (1.001±0.055 and 0.352±0.003, t4w-mRNA=3.052, P>0.05; t4w-protein=3.227, P>0.05), but significantly lower than those in the Dex group (2.351±0.086 and 0.836±0.032) and Ex-NC+ Dex group (2.226±0.484 and 0.888±0.067, F4w-mRNA=296.841, P<0.01; F4w-protein=196.684, P<0.01). The expression levels of Runx2 mRNA and protein in Ex-Lnc+ Dex group (1.176±0.036 and 0.963±0.014) were not significantly different from those in the blank group (1.001±0.077 and 0.934±0.019, t4w-mRNA=2.893, P>0.05; t4w-protein=2.154, P>0.05), but significantly higher than those in the Dex group (0.522±0.007 and 0.501±0.005) and Ex-NC+ Dex group (0.614±0.003 and 0.521±0.008, F4w-mRNA=376.700, P<0.01; F4w-protein=2 239.834, P<0.01). At 8 weeks, the expression levels in all groups were consistent with those at the 4 weeks. Conclusion:The recombinant lncRNA-TCONS_00041960 lentivirus vector can effectively down-regulate the expression of adipogenic gene in the femoral head cells of steroid-induced osteonecrosis of the femoral head (ONFH) model of rats in vivo, and significantly promote the expression of osteogenic gene in the cells at the same time, which can effectively prevent the occurrence of ONFH.

目的:探讨经第2骶椎骶髂(S2AI)螺钉内固定在成人退变性腰椎侧后凸畸形合并骨质疏松症(OP)患者矫形术中的应用效果.方法:收集2016年1月至2019年7月收治的35例成人腰椎侧后凸畸形合并OP患者的资料,男6例,女29例.术中固定至骶2水平(S2AI组)20例,其中男4例,女16例;固定至骶1水平(S1组)15例,其中男2例,女13例.术前、术后第7天及末次随访时,测量侧凸脊柱-骨盆参数,包括侧凸Cobb角、矢状面轴向距离(SVA)、冠状面平衡(C7 PL-CSVL)、骨盆倾斜和局部后凸(RK)Cobb角;并进行疼痛视觉模拟评分(VAS)和Oswe-stry功能障碍指数(ODI)评价.结果:两组患者年龄、手术时长、出血量、随访时间差异均无统计学意义(P>0.05).S2AI组末次随访时侧凸Cobb角(18.0±7.0)°,C7PL-CSVL(6.0±1.8)mm,骨盆倾斜(2.1±1.0)°,RK Cobb角(3.6±17.4)°,SVA(16.9±14.1)mm;与术前比较,上述指标均显著降低(P<0.05).两组患者术后第7天、末次随访时VAS和ODI均较术前降低(P<0.05).两组之间上述指标改善程度比较差异无统计学意义(P>0.05).患者术中均未出现SSEP及MEP信号的改变和丢失.术后随访中未发生退钉、断棒、PJK、DJK及脊柱失代偿.S2AI组术后并发症(螺钉松动和迟发性臀部疼痛)发生率2/20,S1组为11/15,S2AI组并发症发生率低于S1组(P=0.022).结论:应用S2AI螺钉内固定治疗成人腰椎侧后凸畸形合并OP,既能满足强化固定的需要,又可获得有效的矫形效果.