Objective Osteoporosis (OP) is a systemic skeletal disease that increases the risk of fractures by weaking bone. Hypoxia-inducible factor-1α (HIF-1α) plays a crucial role in osteogenesis and osteoblastic differentiation. The purpose of this study is to examine the underlying mechanism of HIF-1α overexpression and its impact on osteoblast development. Methods First, we used the autophagy inhibitor 3-MA in conjunction with either a control lentivirus or an HIF-1α overexpression lentivirus to transfect rat osteoblasts in osteogenic induction media for 3, 7, 14, and 21 days. The effects of HIF-1α overexpression on osteogenic differentiation were evaluated using CCK-8, alkaline phosphatase (ALP) staining, and Alizarin Red staining. Furthermore, we investigated the mechanism by which HIF-1α overexpression mediates autophagy to regulate osteogenic differentiation through immunofluorescence, western blot, and transmission electron microscopy. Ovariectomy (OVX) was performed to establish an osteoporotic rat model. The impact of HIF-1α overexpression on autophagy and bone metabolism was evaluated by locally injecting HIF-1α overexpression lentivirus or control lentivirus, in combination with HE staining, micro-CT, immunohistochemistry, ELISA, western blot, and transmission electron microscopy. Results Overexpression of HIF-1a promotes osteoblast proliferation and enhances ALP staining as well as calcium nodule formation. In addition, the overexpression of HIF-1a significantly increases the relative protein expression levels of osteocalcin (OCN), osteoprotegerin (OPG), HIF-1a, BNIP3, Beclin1, ATG5, and LC3 II/I. This indicates that HIF-1a may facilitate osteoblast differentiation by promoting autophagy. These findings were further corroborated by in vivo experiments, which demonstrated improved pathological morphology in rat femurs, alongside increased bone mineral density (BMD), trabecular thickness (Tb. Th), bone volume/total volume ratio (BV/TV), and trabecular number (Tb. N). Additionally, there was a decrease in trabecular separation (Tb. Sp) and structural model index (SMI), along with upregulated expression of OCN, OPG, HIF-1α, BNIP3, Beclin1, ATG5, and LC3 II. Conclusion HIF-1a overexpression can promote osteogenic differentiation and ameliorate osteoporosis through the induction of autophagy. These insights provide a valuable reference for its potential application in targeted therapy.

Osteoporosis (OP) is a systemic bone metabolic disease. Promotion of osteoblast proliferation and inhibition of cell apoptosis may be helpful for the prevention and clinical treatment of OP. In the current study, we focused on the expression changes and clinical values of lncRNA ROR and miR-145-5p in OP clinical serum samples, and investigated the interactive modulation effect of ROR/miR-145-5p on osteoblast function. Serum samples were obtained from 82 OP patients and 79 healthy individuals. MC3T3-E1 was applied for the cell experiments. Levels of lncRNA ROR and miR-145-5p were detected using qRT-PCR. Transient transfection was performed to regulate gene levels in cells, and cell proliferation and apoptosis were detected. A reciprocal correlation between lncRNA ROR and miR-145-5p was explored. LncRNA ROR was downregulated, and miR-145-5p was overexpressed in OP patients. The combined diagnosis of ROR and miR-145-5p showed good diagnostic value for OP. ROR knockdown promoted the MC3T3-E1 cell apoptosis and inhibited cell proliferation. Luciferase reporting assay verified the target relationship between ROR and miR-145-5p. MiR-145-5p downregulation reversed ROR silence mediated effect on MC3T3-E1 cell proliferation and apoptosis. LncRNA ROR is downregulated and miR-145-5p is highly expressed in OP patients. ROR knockdown may inhibit osteoblast proliferation via targeting miR-145-5p. It may provide a theoretical basis and experimental basis for ROR to be a potential target for the treatment of OP.

背景:最新研究发现氧化应激在骨质疏松症中发挥着重要作用,衰老、雌激素缺乏、核因子E2相关因子2(Nrf2)、血红素加氧酶1与活性氧的产生和骨质疏松发病有着重要的关系.目的:分析左归丸对成骨细胞氧化应激损伤的保护作用机制.方法:实验方案经哈尔滨医科大学附属第二医院伦理委员会批准.分离培养SD乳鼠颅骨成骨细胞,实验分为空白组、模型组(300μmol/L H2O2)、左归丸组(300μmol/L H2O2+10％左归丸含药血清).用H2O2建立成骨细胞氧化应激损伤模型;在成骨细胞氧化应激损伤同时,用左归丸含药血清培养成骨细胞,空白组不做任何处理.测定各组成骨细胞中丙二醛和超氧化物歧化酶含量;Western blot检测成骨细胞中核因子E2相关因子2和血红素加氧酶1蛋白表达.结果与结论:①与空白组比较,模型组中丙二醛含量明显升高,超氧化物歧化酶的含量明显降低;与模型组比较,左归丸组中丙二醛含量明显降低,超氧化物歧化酶的含量明显升高;②与空白组比较,模型组成骨细胞中血红素加氧酶1和核因子E2相关因子2表达明显增加;左归丸组成骨细胞中血红素加氧酶1和核因子E2相关因子2表达明显高于模型组;③结果说明,左归丸对成骨细胞氧化损伤具有保护作用.

Osteoarthritis (OA), a progressive disease of the joints, affects millions of people worldwide. In the present study, we investigated the effects of oridonin, a diterpenoid isolated from Rabdosia rubescens, on IL-1β-induced inflammation using human osteoarthritis chondrocytes. The results showed that oridonin significantly suppressed IL-1β-induced MMP1, MMP3, and MMP13 production. IL-1β-induced NO and PGE2 production, as well as inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression were also attenuated by oridonin. Western blot analysis demonstrated IL-1β-induced NF-κB activation was reduced by oridonin. Furthermore, the expression of PPAR-γ was increased by oridonin in a concentration-dependent manner. PPAR-γ antagonist could reverse the anti-inflammatory activity of oridonin. The results suggested that oridonin could be a candidate agent for the treatment of OA.

Objective To evaluate one-stage repair of leg open fracture with soft tissue defect by fixation plus skin flap.Methods From January 2011 to January 2018,34 patients with leg open fracture plus soft tissue defect were treated at Department of Orthopedics,The Second Affiliated Hospital to Harbin Medical University.They were 31 males and 3 females,aged from 13 to 54 years(average,35 years).According to the Gustilo classification,the defects were type ⅢA in 31 cases and type Ⅲ B in 3 cases.The wound size ranged from 6 cm × 3 cm to 25 cm × 10 cm.All patients were treated with emergency fracture fixation and flap repair by one stage after thorough debridement.Internal bone plating was performed in 32 patients and external fixation at a single arm in 2 patients.All defects were repaired with a free anterolateral thigh flap.Flap survival,vascular crisis and bone infection were followed up postoperatively.Results All the free flaps survived in the 34 cases with no vascular crisis.These patients were followed up for 6 months to 3 years (average,10 years).The flaps healed well,with good appearance and soft texture.No bone infection occurred at the defective sites.After postoperative rehabilitation,the knee extension ranged from 160° to 180°,knee flexion from 90° to 110°,ankle dorsal extension from 90° to 110°,and ankle plantar flexion from 100° to 120°.Conclusion Soft tissue defects in leg open fracture can be effectively treated by flap repair after thorough debridement at the same time of fracture fixation,because the primary wound healing can avoid infection and lead to fine survival of the flap.

Aiyun Liu 1 Jiutao Qiao Liyuan He Zhangmeng Liu Jing Chen 1 Fenghua Pei Yaju Du 1Department of Gastroenterology, The Second Affiliated Hospital of Harbin Medical University, Harbin, People’s Republic of China; 2Department of Orthopedics, The Second Affiliated Hospital of Harbin Medical University, Harbin, People’s Republic of China Background: Colorectal cancer (CRC) is one of the most common malignant tumors in the world. Our previous study revealed that nitrogen permease regulator-like-2 (NPRL2), a promising anti-tumor gene, was downregulated at both the blood and tissue levels in CRC patients compared with that in healthy individuals. Purpose: This study aims to explore the role of NPRL2 in CRC. Methods: Herein, we constructed NPRL2 overexpression lentivirus vectors and transfected them into HT29 cells. The transfected cells were inoculated subcutaneously into nude mice. Tumor growth, pathology, apoptosis, and the protein expression of caspase-3, caspase-7, Bax, Bcl-2, and phosphorylated protein kinase B (p-Akt) were evaluated. To further explore whether NPRL2 could reduce drug resistance of CRC cells against oxaliplatin (L-OHP) and 5-fluorouracil (5-FU), we constructed a tumor model using HT29 cells. The tumor model was treated with lentiviral particles assembled with vectors encoding NPRL2 and exposed to L-OHP and 5-FU. Tumor growth, pathology, apoptosis, and the protein expression of caspase-3, caspase-7, Bax, Bcl-2, p-Akt, P-glycoprotein (P-gp), and multidrug resistance protein 1 (MRP1) were evaluated. Results: The results indicated that in the in vivo CRC xenograft model, NPRL2 reduced the tumor volume and weight and enhanced apoptosis. Our results also confirmed that NPRL2 enhanced the sensitivity of CRC cells to L-OHP and 5-FU. Our studies further demonstrated that NPRL2 exerted anti-tumor and anti-drug resistance effects through the caspase-3, caspase-7, Bax, Bcl-2, Akt, P-gp, and MRP1 pathways. Conclusion: Our present work demonstrated that NPRL2 exhibited anti-tumor effects and enhanced the sensitivities of CRC cells to L-OHP and 5-FU through the P-gp andMRP1 pathways.

目的:探讨NPRL2对人结肠癌细胞自噬的影响,并进一步探讨自噬和凋亡的关系.方法:构建过表达NPRL2慢病毒载体,转染至HT29细胞.检测转染48 h后NPRL2、LC3 B和p62蛋白表达量,吖啶橙染色检测细胞自噬.用自噬抑制剂3-甲基腺嘌呤(3-MA)处理转染后细胞,测定细胞增殖,细胞死亡,细胞caspase-3活性,细胞凋亡和caspase-3、Bax、Bcl-2、LC3 B及p62的蛋白表达量,吖啶橙染色检测细胞自噬.结果:免疫印迹法检测结果表明NPRL2能够激活HT29细胞的自噬.前期研究结果表明,NPRL2能够促进HT29细胞凋亡.进一步研究NPRL2激活的自噬与凋亡之间的关系,结果表明3-MA抑制由NPRL2诱导的自噬,降低细胞活力,抑制其增殖,促进其凋亡,并促进活化caspase-3和Bax的表达,抑制Bcl-2的蛋白表达和细胞自噬.结论:NPRL2能够促进HT29细胞的自噬,但这种自噬抑制了NPRL2引起的细胞凋亡,通过抑制3-MA抑制NPRL2引起的自噬能够有效地促进结肠癌细胞的凋亡.

Glucocorticoids (GCs) are widely used to treat a variety of autoimmune diseases, but long-term use can lead to osteoporosis. To elucidate the mechanism of osteoporosis caused by glucocorticoids and to find effective protective drugs/foods, osteoblasts treated by prednisolone acetate were studied and salvianolic acid B (Sal B) was added to osteoblasts. The results showed that Sal B increased the activity of ALP and stimulated the expression of ALP that had been suppressed by prednisolone acetate. To further study the mechanisms of the protective effect of Sal B on osteoblasts treated with prednisolone acetate, the effects of gene expression involved with bone formation and differentiation were studied. The results show that the mRNA and protein expression of Runx2, Osx, OCN, IGF-I, Col-I and HO-I was up-regulated by Sal B. In conclusion, by stimulating the osteoblast activity and the expression of genes related to bone formation and differentiation, Sal B had a protective effect on osteoblasts that had been treated with prednisolone acetate.

Background: Colorectal cancer (CRC) is one of the most common malignant tumors in the world. Our previous study revealed that nitrogen permease regulator-like-2 (NPRL2), a promising anti-tumor gene, was downregulated at both the blood and tissue levels in CRC patients compared with that in healthy individuals. Purpose: This study aims to explore the role of NPRL2 in CRC. Methods: Herein, we constructed NPRL2 overexpression lentivirus vectors and transfected them into HT29 cells. The transfected cells were inoculated subcutaneously into nude mice. Tumor growth, pathology, apoptosis, and the protein expression of caspase-3, caspase-7, Bax, Bcl-2, and phosphorylated protein kinase B (p-Akt) were evaluated. To further explore whether NPRL2 could reduce drug resistance of CRC cells against oxaliplatin (L-OHP) and 5-fluorouracil (5-FU), we constructed a tumor model using HT29 cells. The tumor model was treated with lentiviral particles assembled with vectors encoding NPRL2 and exposed to L-OHP and 5-FU. Tumor growth, pathology, apoptosis, and the protein expression of caspase-3, caspase-7, Bax, Bcl-2, p-Akt, P-glycoprotein (P-gp), and multidrug resistance protein 1 (MRP1) were evaluated. Results: The results indicated that in the in vivo CRC xenograft model, NPRL2 reduced the tumor volume and weight and enhanced apoptosis. Our results also confirmed that NPRL2 enhanced the sensitivity of CRC cells to L-OHP and 5-FU. Our studies further demonstrated that NPRL2 exerted anti-tumor and anti-drug resistance effects through the caspase-3, caspase-7, Bax, Bcl-2, Akt, P-gp, and MRP1 pathways. Conclusion: Our present work demonstrated that NPRL2 exhibited anti-tumor effects and enhanced the sensitivities of CRC cells to L-OHP and 5-FU through the P-gp and MRP1 pathways.

Liver injury is an important cause of serious liver disease and is characterized by inflammatory and oxidative responses. Esculin, a coumarinic derivative found in Aesculus hippocastanum L., has been shown to exhibit anti-inflammatory and anti-oxidative effects. Here, we investigated the effects and molecular mechanism of esculin on Lipopolysaccharide/D-Galactosamine (LPS/D-Gal)-induced acute liver injury. A mouse model for acute liver injury was induced by intraperitoneal injection with D-Gal and LPS, and was assessed by histology, and serum transaminase analyses. The results showed that esculin significantly reduced the pathological symptoms of acute liver injury, as well as serum AST and ALT levels. LPS/D-Gal-induced liver myeloperoxidase (MPO) activity and malondialdehyde (MDA) content were also suppressed by esculin. Furthermore, LPS/D-Gal-induced liver tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) production were attenuated by esculin. Our data demonstrate that esculin can inhibit nuclear factor kappa B (NF-κB) activation as well as increase nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) expression. In conclusion, this paper demonstrates that esculin protects liver injury induced by LPS/D-Gal via inhibiting inflammatory and oxidative responses.

Background: Osteoporosis has brought about heavy socio-economic burden in the morbidity and medical expenses associated with osteoporosis treatment and various restrictions on behavior of their social roles. Oleanolic acid (OA) is an anti-osteoporosis natural product, but molecular mechanisms of therapeutic effect are not still well known. Purpose: In this study, we explore anti-osteoporosis activity of oleanolic acid and predict the underlying mechanisms by metabolomics strategy. Methods: SD rats were intraperitoneal injection with prednison for once to establish osteoporosis model. Using metabolomics strategy based on ultra-performance liquid chromatography coupled with quadrupole time-offlight/ mass spectrometry (UPLC-TOF/MS), serum samples of 24 rats were analyzed to seek differential metabolites and pathway associated with OA treatment of osteoporosis. In addition, the effect of OA on osteoporosis rats was also evaluated by clinical biochemistry indicators and bone density analysis. Results: Clinical biochemistry indicators and bone density of lumbar and femur were reversed by OA treatment. A total of 25 potential biomarkers were identified in the rats model of glucocorticoid-induced osteoporosis, and oleanolic acid have a regulatory effect on 17 of them that related to some vital metabolic pathway such as linoleic acid metabolism, valine, leucine and isoleucine biosynthesis, phenylalanine, tyrosine and tryptophan biosynthesis as well as cysteine and methionine metabolism. The ingenuity pathway analysis (IPA) platform is applied to further understanding the relationship between metabolic changes and therapeutic effect of OA, which the disordered state carbohydrate metabolism, molecular transport and lipid metabolism in glucocorticoid-induced osteoporosis rats are mainly ameliorated by oleanolic acid. Conclusion: Metabolomics provides a novel method to investigate the anti-osteoporosis effects of OA and probe into the potential mechanisms, and will contributes to the development of new drugs.

Staphylococcus aureus (S. aureus) is a major cause of pneumonia that often affects young and immunocompetent patients. Inflammation plays an important role in the development of S. aureus-induced pneumonia. Geniposide, a major iridoid glucoside component of gardenia fruit, has been reported to have anti-inflammatory and anti-oxidative effects. The purpose of this study was to investigate the protective effects of geniposide on S. aureus-induced pneumonia in mice. Lung histopathological changes were detected by hematoxylin-eosin (H&E) staining. Lung myeloperoxidase (MPO) activity, wet-to-dry (W/D) ratio, and inflammatory cytokine levels in bronchoalveolar lavage fluid (BALF) were measured. The results showed that S. aureus-induced lung histopathological changes were attenuated by geniposide. S. aureus-induced MPO activity and lung W/D ratio were inhibited by treatment of geniposide. Furthermore, the levels of TNF-α and IL-1β in the BALF were also suppressed by geniposide. In addition, geniposide significantly inhibited S. aureus-induced nuclear factor kappa B (NF-κB) activation. Taken together, these results showed that geniposide inhibited S. aureus-induced pneumonia in mice by inhibiting NF-κB signaling pathway. Geniposide might be used as a potential agent for the treatment of S. aureus-induced pneumonia.

Licochalcone A (Lico A), a flavonoid found in licorice root (Glycyrrhiza glabra), has been reported to have anti-inflammatory activity. In this study, we evaluated the anti-inflammatory effects of Lico A on IL-1β-stimulated human osteoarthritis chondrocytes and investigated the possible mechanism. Results demonstrated that Lico A treatment significantly inhibited PGE2 and NO production induced by IL-1β. IL-1β-induced iNOS and COX-2 expression were also inhibited by Lico A. Lico A inhibited MMP1, MMP3, and MMP13 production in IL-1β-stimulated chondrocytes. Lico A also inhibited IL-1β-induced phosphorylation of NF-κB p65 and IκBα. Meanwhile, Lico A was found to upregulate the expression of Nrf2 and HO-1. However, Nrf2 siRNA reversed the anti-inflammatory effects of Lico A. In conclusion, our results suggested that Lico A showed anti-inflammatory effects in IL-1β-stimulated chondrocytes by activating Nrf2 signaling pathway.

Pneumonia is the leading cause of death in infants and young children. Staphylococcus aureus (S.aureus) is one of the most important bacteria that leads to pneumonia. Tenuigenin (TGN), a major active component isolated from the root of the Chinese herb Polygala tenuifolia, has been known to have anti-inflammatory effect. In this study, we aimed to investigate the protective effects of TGN on S.aureus-induced pneumonia in mice. The results showed that TGN significantly attenuated S.aureus-induced lung histopathological changes. TGN also inhibited lung wet/dry (W/D) ratio, and inflammatory cytokines TNF-α and IL-1β production. Furthermore, S.aureus-induced NF-κB activation was significantly inhibited by the treatment of TGN. In conclusion, the results of this study showed that TGN protected against S.aureus-induced pneumonia by inhibiting NF-κB activation. TGN might be a potential agent in the treatment of pneumonia induced by S.aureus.

目的:探讨中国汉族人群中载脂蛋白Eε4等位基因与急性脊髓损伤(ASCI)预后的相关性.方法:采用限制性片段长度多态性聚合酶链反应(PCR-RFLP)方法,检测2012年5月-2015年10月就诊于哈尔滨医科大学附属第二医院脊柱外科的50例(男性患者35例,女性患者15例)颈段(C3-C8)脊髓损伤的中国汉族患者载脂蛋白E ε4等位基因的遗传学状态,并以ASIA脊髓损伤神经学分类标准为患者神经功能恢复情况进行评估,分析中国汉族人群中载脂蛋白Eε4等位基因与急性脊髓损伤预后的相关性.结果:在运动方面,载脂蛋白Eε4阴性的患者恢复情况显著强于载脂蛋白Eε4阳性的患者恢复情况(mean 6.1vs 3.7,P=0.04).而在针刺觉方面,载脂蛋白Eε4阴性的患者恢复情况显著弱于载脂蛋白Eε4阳性的患者恢复情况(mean 4.0 vs 6.1,P=0.03).在轻触觉方面,载脂蛋白Eε4阴性和阳性患者的恢复情况没有显著的差异(mean 7.4 vs 5.9,P=0.28).结论:中国汉族人群中载脂蛋白Eε4等位基因与ASCI的预后密切相关:载脂蛋白Eε4阳性不利于患者运动的恢复,利于患者针刺觉的恢复.

Thymoquinone, an active ingredient isolated from Nigella sativa, has been reported to have anti-inflammatory effects. However, the anti-inflammatory effect of thymoquinone on IL-1β-stimulated osteoarthritis chondrocytes remains unclear. In this study, we designed to investigate the anti-inflammatory effects and elucidated the underlying mechanism of thymoquinone on IL-1β-stimulated human osteoarthritis chondrocytes. The effects of thymoquinone on inflammatory mediators COX-2, iNOS, NO, PGE2, as well as MMP-1, MMP3, MMP13 production were detected. The results demonstrated that thymoquinone concentration-dependently inhibited IL-1β-induced COX-2, iNOS, NO, and PGE2 production. Thymoquinone also suppressed IL-1β-induced MMP-1, MMP3, and MMP13 production. We found that thymoquinone significantly inhibited IL-1β-induced NF-κB activation and IκBα degradation. In addition, thymoquinone was found to suppress IL-1β-induced mitogen-activated protein kinases (MAPKs) activation. In conclusion, thymoquinone inhibited IL-1β-induced inflammatory mediator production by inhibition of NF-κB and MAPKs signaling pathways in osteoarthritis chondrocytes. Thymoquinone may be a potential agent in the treatment of osteoarthritis.

The organotypic spinal cord slice is a technology which culture and study spinal cord slice by organotypic culture in vitro,living slicing machine and microporous membrane.The characteristics of the technology is simple operation,visual observation,long time experiment in vitro and facilitating applied experimental factors.The method provide more technical support and new ways for studying on pathology and physiology of spinal cord in vitro,but the application and report of the technology in domestic is few.Because its high application value,this paper makes a review on development,characteristic,announcement,and application of organotypic spinal cord slice.

Acute spinal cord injury(ASCI) is a serious and common disease of the orthopedics,After injury neurological recovery and reconstruction is the hot spot of the research in recent years,and the research of cell transplantation get wide attention and make great progress.This paper introduces the present research status of the cell transplantation for the treatment of spinal cord injury.Among them,the source of the cell transplantation,the timing of the cell transplantation,the approach of the cell transplantation,the problem of survival after cell transplantation and the coping strategy are maked a detailed description.At the same time we make a simple review for the preconditioning method which increase the survival rate of the cell transplantation.Many new therapeutic measures are still in the experimental stage at present,but the transform from the successful empirical study of neuroprotection and neurotization to clinical application by some new skills will come soon.

Objective: To investigate the effect of glial cells' apoptosis and spinal-vesicle's forming on neural stem cells(NSCs) by hypoxia preconditioning in rats with acute spinal cord injury(ASCI).Methods: Thirty rats are divided into five groups: sham-operated group,spinal cord injury group,DFO group,routine cultured NSCs group and hypoxic preconditioning cultured NSCs group.The method of improved Allen's hit was used to produce the ASCI model.Pathology tissue was took for 7d after operation,TUNEL was used to de-tect glial cells' apoptosis,and HE staining was used to detect spinal-vesicle.Results: There were no obvious morphological changes be-tween using DFO cultured NSCs and routine condition cultured NSCs.The number of TUNEL-positive cells increased and spinal-vesi-clese formed significantly in the damage zone after spinal cord injuryed.Comparing with the routine condition cultured group,hypoxic preconditioning cultured NSCs reduce the number of the apoptosis nerve cells in the injured spine and reduce the area of spinal-vesicle after transplantation obviously.Conclusion: Transplanting the hypoxic preconditioning cultured NSCs can restraint glial cells' apoptosis,and reduce the area of spinal-vesicle.