OBJECTIVE To investigate the ameliorative effect and mechanism of Euphorbia hirta L.-derived exosome-like nanovesicles（Eh-ENVs） on acetaminophen （APAP）-induced liver injury based on the nuclear factor erythroid 2 related factor 2 （Nrf2）/heme oxygenase-1 （HO-1）/NAD（P）H：quinone oxidoreductase 1 （NQO1） pathway. METHODS The safety of Eh-ENVs was evaluated by examining their effects on the viability of RAW264.7 and AML12 cells， as well as serum liver and kidney function indicators and histopathology of liver， lung， and other tissues in normal mice. A lipopolysaccharide （1 μg/mL）-induced RAW264.7 cell inflammation model was constructed to investigate the effects of 10 and 20 μg/mL Eh-ENVs on the mRNA expression of inflammatory factors and reactive oxygen species （ROS） level in model cells， and the uptake efficiency of Eh-ENVs by RAW264.7 cells was also examined. An APAP-induced liver injury mouse model was established to investigate the effects of 4 mg/kg Eh-ENVs on serum liver function indicators， liver histopathology， mRNA expression of inflammatory factors， malondialdehyde （MDA） level， superoxide dismutase （SOD） level， and mRNA and protein expressions related to the Nrf2/HO-1/NQO1 pathway in liver tissue of model mice. RESULTS In vitro results showed that Eh-ENVs had no inhibitory effect on the proliferation of RAW264.7 and AML12 cells；Eh-ENVs could be efficiently taken up by RAW264.7 cells and significantly reduced the mRNA expression of interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α）， and ROS level in cells （ P ＜0.05）. In vivo results showed that 4 mg/kg Eh-ENVs had no obvious toxic side effects on normal mice，could significantly decrease the serum alanine transaminase （ALT） and aspartate transaminase （AST） levels in model mice （ P ＜0.05），upregulated/increased the mRNA expressions of IL-10， as well as the mRNA and protein expressions of Nrf2， HO-1， and NQO1， and SOD level in liver tissue （ P ＜0.05）， and down-regulated/decreased the mRNA expression of TNF-α， IL-1β and MDA level in liver tissue （ P ＜0.05）. CONCLUSIONS Eh-ENVs may activate the Nrf2/HO-1/NQO1 pathway to inhibit inflammatory response and alleviate oxidative stress， thereby improving APAP-induced liver injury.

BACKGROUND:Irreducible femoral neck fracture was difficult to obtain anatomic reduction.As a new type of internal fixation,the femoral neck system is still blank for the treatment of non-anatomical reduced femoral neck fractures. OBJECTIVE:To explore the biomechanical stability of femoral neck system internal fixation under nonanatomical reduction in the treatment of femoral neck fractures based on finite element analysis. METHODS:CT data of the hip joint of a healthy female adult were obtained.Anatomical reduction of femoral neck fracture models with Pauwels angles of 30°,50°,and 70° were established using Mimics 21.0,Geomagic Wrap 2021,and SolidWorks 2020.The fracture proximal ends of the three anatomical reduction models were shifted upward by 2 mm along the fracture line,and three positive buttress models with different Pauwels angles were obtained.In the same way,three negative buttress models were acquired by shifting downward by 2 mm.SolidWorks 2020 was used to make the femoral neck system internal fixation,and the nine femoral neck fracture models were assembled with the femoral neck system.Then Ansys 19.0 was used for finite element analysis.The displacement distribution and maximum displacement,stress distribution and maximum stress of the femur and femoral neck system were recorded under 2100 N stress. RESULTS AND CONCLUSION:(1)When Pauwels angles were 30°,50°,and 70°,the maximum stresses of the femoral neck system appeared to be concentrated at the junction of the sliding hip screw and anti-rotation screw.The maximum femur stresses appeared to be concentrated in the medial cortex of the femur.The maximum displacement was concentrated at the upper of the femoral head and femoral neck system.(2)When Pauwels angles were 30° and 50°,the maximum displacement and maximum stress of the femoral neck system and femur were:negative buttress>anatomical reduction>positive buttress.(3)When Pauwels angle was 70°,the maximum displacement and maximum stress of the femoral neck system were:negative buttress>anatomical reduction>positive buttress;the maximum displacement and maximum stress of the femur were:negative buttress>positive buttress>anatomical reduction.(4)With the increase of Pauwels angle,the biomechanical advantage of the positive buttress was weakening.However,it was better than a negative buttress.When Pauwels angle was 30°,positive buttress was more stable than anatomical reduction.When Pauwels angle was 50°,the biomechanical difference between positive buttress and anatomical reduction became smaller.When Pauwels angle was 70°,the stability of anatomical reduction was slightly better than positive buttress.(5)If it was difficult to achieve anatomical reduction of femoral neck fracture during operation,but the positive buttress had been displaced within 2 mm,the femoral neck system could be used to offer stable mechanical fixation.It is necessary to avoid negative buttress reduction.

OBJECTIVE To study the effects of bergapten in the treatment of liver fibrosis and its mechanism based on serum metabolomics. METHODS Forty mice were divided into normal control group （0.5% carboxymethyl cellulose sodium solution）， model group （0.5% carboxymethyl cellulose sodium solution）， and BP low-dose and high-dose groups （50， 100 mg/kg）， with 10 mice in each group. Except for the normal control group， the other three groups were all treated with carbon tetrachloride to induce liver fibrosis model； they were given relevant medicine/solution intragastrically， once a day， for consecutive 8 weeks. After the last medication， the levels of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） in serum were detected， and liver pathological changes were observed； the expressions of α-smooth muscle actin （α-SMA） and Collagen Ⅰ were detected in liver tissue； the serum of the mice was collected for metabolomics analysis. RESULTS Compared with the model group， serum levels of ALT and AST and protein expressions of α-SMA and Collagen Ⅰ in liver tissue were decreased significantly in BP high-dose and low-dose groups （P＜0.05）， while liver fibrosis was improved significantly. Meanwhile， metabolomics analyses showed that there were a total of 175 serum differential metabolites in the BP high-dose group and model group， of which 18 substances were upregulated and 157 substances were downregulated； the main metabolic pathways involved in bergapten intervention were pyrimidine metabolism， butanoate metabolism， fatty acid synthesis， tyrosine metabolism， β-alanine metabolism， nicotinic acid and nicotinamide metabolism， glutathione metabolism， etc. CONCLUSIONS BP is effective in the treatment of liver fibrosis by regulating pyrimidine metabolism， butanoate metabolism， glutathione metabolism and so on in rats with liver fibrosis.

Two patients (patient 1, a 63-year-old female; patient 2, a 22-year-old male) were treated with anlotinib for lung adenocarcinoma and synovial sarcoma, respectively. Patient 1 developed cough, expectoration, and dyspnea after 8 cycles of anlotinib treatment (12 mg orally once daily, 2 weeks of treatment and 1 week of withdrawal as a treatment cycle). Spontaneous pneumothorax was diagnosed by chest CT. The spontaneous pneumothorax was considered to be related to anlotinib. Then the drug was stopped. After thoracic cavity closed drainage therapy, the patient′s dyspnea was relieved. Due to the progress of the disease, the patient received reduced-dose anlotinib treatment (10 mg/d, with the same usage as before). Spontaneous pneumothorax recurred in the first cycle. Anlotinib treatment was terminated and the symptomatic treatments including thoracic cavity closed drainage, expectorant, anti-infection, and nutritional support were given. Ten days later, chest X-ray examination showed that pneumothorax was cured. Patient 2 developed spontaneous pneumothorax (found by chest CT examination) after 9 cycles of treatment with anlotinib (12 mg/d, with the same usage as that in patient 1). But the patient had no obvious discomfort and continued to use anlotinib under close monitoring of respiratory status.

Two patients (patient 1, a 63-year-old female; patient 2, a 22-year-old male) were treated with anlotinib for lung adenocarcinoma and synovial sarcoma, respectively. Patient 1 developed cough, expectoration, and dyspnea after 8 cycles of anlotinib treatment (12 mg orally once daily, 2 weeks of treatment and 1 week of withdrawal as a treatment cycle). Spontaneous pneumothorax was diagnosed by chest CT. The spontaneous pneumothorax was considered to be related to anlotinib. Then the drug was stopped. After thoracic cavity closed drainage therapy, the patient′s dyspnea was relieved. Due to the progress of the disease, the patient received reduced-dose anlotinib treatment (10 mg/d, with the same usage as before). Spontaneous pneumothorax recurred in the first cycle. Anlotinib treatment was terminated and the symptomatic treatments including thoracic cavity closed drainage, expectorant, anti-infection, and nutritional support were given. Ten days later, chest X-ray examination showed that pneumothorax was cured. Patient 2 developed spontaneous pneumothorax (found by chest CT examination) after 9 cycles of treatment with anlotinib (12 mg/d, with the same usage as that in patient 1). But the patient had no obvious discomfort and continued to use anlotinib under close monitoring of respiratory status.

Objective To investigate the difference of blood supply in different areas of the normal femoral head in adults.Methods A retrospective study analyzed the DCE-MRI parameters data of 2 5 normal adults (25 right hips)in our hospital.The femoral head was assumed to be a sphere.On T1 WI of DCE-MRI,the right femoral head was divided into three parts equally in coronal,sagittal, and transverse plane respectively.Time to peak(TTP),maximum of enhancement at first past(fEmax),maximum of enhancement (Emax)were measured in three parts and compared in coronal,sagittal and transverse plane respectively.Results (1 )In coronal plane,there were no significant differences between the three parts in TTP,fEmax and Emax.(2 )In transverse plane,fEmax and Emax were lower in the transverse superior zone than that in the transverse central zone significantly (P＝0.024 and P＝0.001)and the transverse inferior zone(P＝0.005 and P＝0.018).TTP was higher in the transverse superior zone than that in the transverse central zone and the transverse inferior zone(P＝0.005 and P＝0.018).(3)In sagittal plane,fEmax and Emax were lower in the sagittal lateral zone than that in the sagittal centralzone (P＝0.019 and P＝0.041)and sagittal medial zone (P＝0.047 and P＝0.01 1).TTP was higher in the sagittal lateral zone than that in the sagittal central zone and the sagittal medial zone(P＝0.014 and P＝0.010).There were no significant differences between other paired zones of the any values.Conclusion DCE-MRI can show the microcirculation in the normal adult femoral head in a non-invasive way.According to DCE-MRI parameters,there are poor perfusions on the transversesuperior and sagittallateral zones of the normal femoral head in adults.

Objective: To investigate the early diagnosis and treatment options for steroid-induced osteonecrosis of the femoral head (SONFH). Methods: Retrospective analysis was conducted in 73 patients with unilateral SONFH from January 2010 to June 2017. There were 27 males and 46 females, aged 34.26±10.35 years (range, 20-45 years). The following type of cases were included, 21 cases of ARCO I, 24 cases of IIa, 8 cases of IIb, 18 cases of IIc and 2 cases of IIIa. Patients with ARCO I were limited in weight and were treated with low molecular weight heparin sodium and ginkgo dharma drugs after excluding bleeding risk. Core decompression was performed in patients with ARCO IIa and IIb vascularized greater trochanter bone transfer was conducted in patients with IIc and IIIa. During the follow-up duration, when the ARCO I progressed to ARCO IIa and IIb, the core decompression was performed. When the ARCO IIa and IIb progressed to the ARCO IIc or IIIa, the vascularized greater trochanter bone transfer was conducted. Vascularized greater trochanter bone combined with the tantalum rod was implanted in ARCO IIIb. The femoral head reconstruction with the larger trochanter bone flap or joint replacement was performed in ARCO IIIc and above. The Harris hip score was used to evaluate the clinical outcomes. The femoral head survival curve of SONFH was drawn with the end of joint replacement. Results: All 73 patients were followed up for an average of 29 months (range, 18 to 48 months). After treatment of 21 patients with ARCO I, a total of 14 (67%) patients had no ARCO staging progress. After 6 months, three patients progressed to ARCO IIb and 4 patients progressed to ARCO IIc. All 32 patients with ARCO IIa and IIb were treated, of which 25 (78%) did not progress but 4 patients progressed to ARCO IIIa at 6 months. Only one patient progressed to ARCO IIIb at 6 months, and 2 patient progressed to ARCO IIIc at 12 months. There were 18 patients with ARCO IIc underwent treatment, of which 14 patients (78%) did not progress but 2 patients progressed to ARCO IIIa at 6 months and 2 patients progressed to ARCO IIIc at 24 months and to ARCO IV at 42 months, respectively. Two patients of ARCO IIIa, 1 patient did not progress and the other one progressed to ARCO IV at 36 months. The Harris hip score was 90.48±5.36 before treatment and 91.76±8.19 at the last follow-up. The difference was not statistically significant (t=1.231, P=0.402). The Harris hip score of patients with ARCO IIa and IIb in creased from 75.22±8.30 to 84.47±16.77 with statistically significant difference (t=5.624, P<0.001). The Harris hip score of patients with ARCO IIc and IIIa increased from 59.80±9.24 to 77.75±20.75 with statistically significant difference (t=12.033, P<0.001). Four patients eventually underwent arthroplasty. The femoral head survival rate was 98.6% at 2 years and 94.5% at 4 years. Conclusion The SONFH can be diagnosed early by screening, and is treated by different reserving hip methods according to severe stages. The short-term clinical effects are satisfied.

BACKGROUND:There are a variety of treatments for femoral head necrosis,but their efficacy is not confirmed and unified.How to improve the differentiation ability of osteoblasts in the femoral head and improve the biomechanical support after the repair of the femoral head is an urgent problem to be solved.OBJECTIVE:To explore the clinical outcome of stem cells combined with vascularized iliac bone flap and tantalum rod implantation for the treatment of osteonecrosis of the femoral head (ONFH).METHODS:Totally 28 cases (36 hips) of non-traumatic ONFH admitted at the Zhongshan Hospital of Dalian University from January 2010 to January 2011 were enrolled.Bone marrow samples were extracted from each patient to isolate bone marrow stromal stem cells which were cultured in vitro for 2 weeks.Tantalum rod implantation with vascularized iliac bone graft was conducted to restore the femoral head shape,and then,prepared stem cell suspension were injected into the iliac bone flap and into the subchondral space of the femoral head.RESULTS AND CONCLUSION:All the 28 cases (36 hips) were followed up for 6-20 months (average 12 months),and their Harris hip scores and visual analogue scale scores at postoperative 6 and 12 months were significantly higher than the baseline (P ＜ 0.05).The Harris hip score at postoperative 12 months was significantly higher than that at postoperative 6 months (P ＜ 0.05),but there was no significant difference in the visual analogue scale scores at 6 and 12 months postoperatively (P ＞ 0.05).At the end of 12-month follow-up,clinical outcomes were excellent in 13 hips,good in 15 hips,fair in 4 hips,and poor in 4 hips,with an excellent and good rate of 90％.These findings indicate that autologous bone marrow stromal stem cell transplantation with vascularized iliac bone flap and tantalum rob implantation is an effective method with high clinical success rate for the treatment of ONFH.

BACKGROUND: The imaging methods of evaluating the volume of necrotic femoral head are based on Kerboul angle and necrosis index revealed on X-ray, as well as CT and MRI three-dimensional reconstruction, but the accuracy of each method is different, and none is widely accepted. Additionally, the association of the volum and degree of necrotic femoral head with the status of blood supply to the femoral head remain unclear. OBJECTIVE: To evaluate the accuracy of each imaing method for assessing different stages of femoral head necrosis, and to confirm a convenient method used for measuring the volum of necrotic femoral head. METHODS: This diagnostic in vivo animal trial was finished in the Affiliated Zhongshan Hospital of Dalian University, Liaoning Province, China. The canine models of steroid-induced necrosis of femoral head were prepared, and received CT and MRI examinations, followed by three-dimensional reconstruction combined with Micro-CT to make accuratediagnosis; the degree of necrotic femoral head and its association with the status of blood supply were evaluated throughangiography and histopathological examination. All above outcome measures were compared with the Association Research Circulation Osseous to testify the consistency of this precise method with traditional methods. The experiment follows the national guidelines for the Care and Use of Laboratory Animals, and Consensus author guidelines on animal ethics and welfare by the international Association for Veterinary Editors (IAVE). The article was prepared in accordance with the Animal Research: Reporting of In Vivo Experiments Guidelines (ARRIVE Guidelines). RESULTS AND CONCLUSION: The aim of the study is to obtain a accurate measurement method by detecting different stages of osteonecrosis using different imaging techniques, which enables surgeons make exact diagnosis and clinical stage of osteonecrosis, thus desiging standardized treatment program.

Objective To investigate the possibility of the domestic reticulated vitreous carbon as a kind of scaffold material for bone tissue engineering,the biocompatibility of domestic reticulated vitreous carbon was first successfully tested with bone marrow stromal cells (BMSCs) in vitro and for bone tissue repair in vivo.Methods From June,2013 to August,2014,the morphology and proliferation of BMSCs co-cultured with scaffold material in vitro was measured.Differences of measurement were compared with single factor analysis of variance to detect the cytotoxicity of reticulated vitreous carbon.In vivo reticulated vitreous carbon were implanted into the bone defect site and the groin.After 12 weeks,the biocompatibility of reticulated vitreous carbon was observed.Results MTT results showed that after 7d co-culture,the survival and proliferation of BMSCs had not been significantly inhibited (P ＞ 0.05).Inverted fluorescence microscope and scanning electron microscope found that newly developed three-dimensional domestic reticulated vitreous carbon could promote adhesion,aggregation and proliferation of BMSCs in vitro.Studies in vivo demonstrate that implanted reticulated vitreous carbon with a high porosity and host bone may produce a stable connection and integration.Conclusion Non-cytotoxic domestic reticulated vitreous carbon can promote the adhesion and proliferation of bone marrow mesenchymal stem cells in vitro and has good bone induction properties in vivo.