Objective To investigate the effect of over-expression of ROBO4 on permeability of human renal glomerular endothelial cells (HRGECs) in high glucose medium.Methods HRGECs infected with recombinant lentiviral vector ROBO4 were cultured in high glucose or low glucose medium in vitro.The protein levels of ROBO4 and ARF6 in each group were detected by Western blotting.The endothelial permeability was measured by the effiux of fluorescein isothiocyanate-dextran (FITC-Dextran)permeated through the monolayer endothelial cells using Transwell cell model system.The cell viability after lentivirus transfection was measured by CCK8 assay.Results The transfection rate of lentiviruses in HRGECs reached 80％ 72h after,and obvious overexpression of ROBO4 protein was in transformed cells compared with the empty vector group (P＜0.05).The lentivirus-mediated ROBO4 transfection did not affect cell viability of HRGECs.Compared with the low glucose group,the expression of ROBO4 increased obviously after 12h,but declined after 24h (P＜0.05),and reached to minimun after 72h (P＜0.05).On the contrary,the expression of ARF6 increased after 12h,and the increase reached to the maximum after 72h (P＜0.05).Furthermore,the vascular permeability increased gradually after 24h,and reached to the maximum after 72h (P＜0.05) in high glucose group.Compared with the empty vector group,the over-expression of ROBO4 inhibited the expression of ARF6 significantly,and the FITC-Dextran permeability reduced obviously.Conclusion Over-expression of ROBO4 may significantly enhance the barrier functions of HRGEC in high glucose medium,and ROBO4 activation may be a potential therapeutic approach in diabetic nephropathy.

Objective To prepare phospholipid-based sulfur hexafluoride gas-filled microbubbles, which were designed for targeting microbubbles. Methods The microbubbles composed of DSPC and mPEG2000-PE were produced by film-sonication method,and were compared with SonoVue. In the physical chemistry characteristics studies, the morphology, particle diameter, concentration, pH value and osmotic pressure were investigated. In the echogenicity studies, contrast harmonic imaging technique was used to investigate the enhancement of vitro water sac and normal rabbit kidney parenchyma. Results The lipid microbubbles were well-distributed, round with air holes. The average diameter of self-made microbubbles and SonoVue were 2.25 μm and 2.50μm respectively. The average diameter of self-made microbubbles and SonoVue distributed from 0.4 μm to 10 μm and 0.2 μm to 10 μm,and 90% were under 6 μm and 8 μm respectively,the concentration were 5 x 108～10 x 108/ml and 1 x I08～5 x 108/ml respectively with a stability of 6 hours. In vitro water sac, the gray scale of self-made lipid microbubbles and SonoVue were 121.67±6.76 and 122.33 ± 4.53 respectively( P>0.05). In normal rabbit kidney parenchyma, the peak video intensity of normal rabbit kidney parenchyma of self-made lipid microbubbles and SonoVue were 72.00 ± 7.21 and 74. 65± 10.93 respectively(P>0.05). Conclusions The lipid microbubbles have satisfactory physical chemistry characteristics and echogenicity.

Objective To summarize the first experience with ultrasound-guided percutaneous ab lation treatment (PAT) for recurrent hepatoblastoma (HB) after liver resection in children.Methods From August 2013 to April 2015,PAT was used to treat 6 children with a total of 9 recurrent HB,including 5 patients with 8 tumors in the liver and 1 patient with 1 tumor in the lung.The mean size of ablated tumors was (1.5 ± 0.8) cm,and the tumor size range was 0.7 cm to 3.1 cm.Results Four patients were performed percutaneous radiofrequency ablation (RFA) for recurrent HB;and 2 patients were performed percutaneous ethanol injection (PEI).Ablation success was achieved in all patients (6/6,100％).The complete ablation rate after the first ablation session was 88.9％ (8/9) on a tumor-by-tumor basis.Only 1 patient developed a fever with temperature ＞ 39 ℃;it was resolved by conservative therapy.During the follow-up period of 5-30 months,3 patients died to tumor progression.The 1-and 2-year overall survival rates after ablation were 83.3％ and 41.7％,respectively.Conclusions PAT is a safe and promising therapy for children with recurrent HB after liver resection,and further investigation in large-scale randomized clinical trials is required to determine its role in the treatment of this disease.

BACKGROUND:Mimics software is a three-dimensional (3D) image processing and editing tool based on CT scan data. Mimics software can rebuild the data and images gotten in CT, MRI and ultrasound scans into 3D images and display on the computer screen so as to help clinicians understand the type of fracture and the relationship of the 3D structure of the surrounding tissue and to provide a great help in the development of orthopedics operation. OBJECTIVE:To explore the application effect of Mimics software 3D reconstruction on perioperative period of Pilon fracture. METHODS:This study selected 61 cases of Pilon fracture, who received the surgery in the Zhongshan City Xiaolan People’s Hospital from September 2008 to September 2013, as research objects. They were randomly divided into 3D group and control group in accordance with the time of admission. Al patients underwent anterioposterior and lateral X-ray film examination and multi-slice spiral CT scan. Patients in the control group received internal fixation according to above examination results. Patients of the 3D group, on the base of those of the control group, were subjected to internal fixation after three-dimensional entity reconstruction by using Mimics V 10.0 software. Operation time and functional curative effect in the postoperative folow-up were compared between the two groups. RESULTS AND CONCLUSION:The operation time was significantly less in the 3D group than in the control group (P < 0.05). The number of patients with excelent 6-month functional curative effect was more, and the number of patients with poor effect was less in the 3D group than in the control group. The 6-month functional curative effect was better in the 3D group than in the control group (P < 0.05). These results showed that the application of Mimics software 3D reconstruction to the perioperative period of Pilon fractures can give comprehensive assessment of the situation of fracture, optimize and improve the preoperative plan and reduce the risk of surgery, and promote the successful completion of internal fixation.

Objective To analyze the factors related to the occurrence of complications in performing CT-guided percutaneous lung biopsy with 18 G-Bard Magnum needle. Methods CT-guided percutaneous lung biopsy with 18 G-Bard Magnum needle was carried out in a total of 58 patients. The postoperative complications were recorded, and the related factors causing complication were analyzed. Results Successful puncturing was achieved in all patients with a technical success rate of 100%. Postoperative complications included pulmonary hemorrhage (n=11,19.0%), a little amount of bleeding in needle tract (n=7,12.1%), hemoptysis (n=3,5.2%), hemothorax (n=1,1.7%), and pneumothorax (n=10,17.2%). Chi-square test showed that the occurrence of pulmonary hemorrhage bore a close relationship to the lesion’s diameter, the distance between the lesion and the chest wall, the lesion’s location and times of puncturing (P<0.05). The occurrence of pneumothorax was closely correlated with the age, the distance between the lesion and the chest wall, the presence of perifocal emphysema, the lesion’s location and times of puncturing (P<0.05). Univariate analysis indicated that the postoperative complications were liable to occur in the patients whose imaging examination showed perifocal emphysema and lung hilar lesion, and who had more than two independent risk factors (P<0.05). Conclusion CT-guided percutaneous lung biopsy with 18 G-Bard Magnum needle is an accurate and safe technique with relatively higher positive diagnosis rate, but this technique should be carefully used in patients who has perifocal emphysema, or lung hilar lesion, or more than two independent risk factors.(J Intervent Radiol, 2015, 24:792-796)

Objective To investigate the factors affecting Fly-Thru (FT) and the best reconstruction conditions in a bile duct model.Methods The bile duct model was made with corn powder and rubber pipes.FT data of bile duct in vitro were acquired with a 3-dimensional probe of Toshiba Aplio 500,and the data were reconstructed offline in Fly-Thru workstation.Two US doctors assessed the FT images in three degrees.Results The images were divided into good,moderate and poor groups with corresponding score 3,2,1.Of 162 images,72 were rated as grade 1,57 as grade 2 and 33 as grade 3.According to x2 analysis,the diameter,scanning way,and Filter were brought into multinomial regression,then the scanning way was found to be correlated to FT images quality as well as filter (likelihood ratio =87.598,294.290,x2 =9.492,313.722,respectively,P ＜0.01).Best images of FT can be acquired by transverse scanning,reconstructing in Filter =3,gain =5(0,dynamic range (DR) =50 and threshold ranging 40-90.Conclusions Best FT images in the model could be achieved by scanning in short-axis,and reconstructing the data in Filter =3,gain =50,DR =5(0 and threshold ranging 40-90,which would be basic information for advanced clinic study.

BACKGROUND:With the emergence of more sophisticated imaging instruments in recent years, orthopedic preoperative examination becomes more clear and accurate. With the help of three-dimensional reconstruction technology, it is bound to improve the accuracy of pedicle screw placement. OBJECTIVE:To investigate the accuracy and safety of digital three-dimensional reconstruction technique in the placement of thoracolumbar pedicle screw. METHODS:200 cases of pedicle screw fixation were col ected and randomly divided into two groups. One group was treated with traditional anatomical landmarks combined with intraoperative fluoroscopic fixation of pedicle screws. Assisted thoracic pedicle screws were implanted with digital three-dimensional reconstruction technique. Preparation time, the amount of bleeding, accuracy of pedicle screw placement, and neurovascular injury of pedicle were recorded by comparing the preoperative and postoperative spine digital models of the pre-and post-virtual spine. RESULTS AND CONCLUSION:(1) The accuracy rate of pedicle screw placement was 85%and 99%in the nail-hand nail group and the auxiliary nail group respectively, and the success rate of one-time placement was 80%and 95%;the excel ent and good rates of placement were 86%and 96%, respectively (P>0.05). (2) The preparation time of nail channel was (5.87±1.34) minutes and the volume of blood loss during nailing was (10.08±7.58) mL in the nail-hand nailing group. The preparation time of nail channel was (3.91±2.28) minutes and the volume of blood loss during nailing was (18.31±8.56) mL in the auxiliary nail group. There was significant difference between the two groups (P<0.05). (3) Nerve injury occurred in six cases, and vascular injury in eight cases in the nail-hand nailing group. Nerve injury occurred in 0 case, and vascular injury in one case in the auxiliary nail group. There were significant differences in nerve and vascular injuries between the two groups (P<0.05). (4) These results confirm that the accuracy and safety of digital three-dimensional reconstruction technique in the treatment of thoracolumbar pedicle screw placement are high.

Objective:To investigate whether astragaloside IV (AS-IV) exerts neuroprotective effects via zinc ion (Zn 2+) modulation of mitochondria-associated endoplasmic reticulum membranes (MAMs) and to elucidate the possible mechanisms. Methods:PC12 cells (rat adrenal pheochromocytoma cells) were routinely cultured; they were divided into 7 groups: control group (routinely cultured), 2-DG group (treated with 2-DG at 50 μmol/L for 30 min), astragaloside IV+2-DG group (treated with 50 μmol/L astragaloside IV for 20 min, and then treated with 50 μmol/L 2-DG for 30 min), astragaloside IV group (treated with 50 μmol/L astragaloside IV for 20 min), N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine (a zinc chelator, TPEN)+astragaloside IV+2-DG group (treated with 10 μmol/L TPEN for 10 min, 50 μmol/L astragaloside IV for 20 min, and then 50 μmol/L 2-DG for 30 min); TPEN group (treated with 10 μmol/L TPEN for 10 min), TPEN+2-DG group (treated with 10 μmol/L TPEN for 10 min and then treated with 50 μmol/L 2-DG for 30 min). The expressions of glucose-regulated protein (GRP)78, GRP94, cysteine-containing aspartate proteolytic enzyme caspases-8, B cell receptor associated protein 31 (BAP31) and mitochondrial fission protein 1 (Fis1) were detected by Western blotting. Annexin V-FITC/PI kit was used to detect the apoptosis level. Immunofluorescence was used to detect the Fis1 protein expression, and mitochondrial fluorescence probe TMRE was used to detect the opening of mitochondrial permeability transition pore (mPTP).Results:(1) Compared with the control group, the 2-DG group had significantly increased expressions of GRP78, GRP94, Caspase-8, Bap31 and Fis1, and green fluorescent intensity of Annexin V-FITC ( P<0.05); compared with the 2-DG group, the astragaloside IV+2-DG group had significantly decreased expressions of GRP78, GRP94, Caspase-8, Bap31 and Fis1, and green fluorescent intensity of Annexin V-FITC ( P<0.05); compared with the astragaloside IV+2-DG group, the TPEN+astragaloside IV+2-DG group had significantly increased expressions of GRP78, GRP94, Caspase-8, Bap31 and Fis1, and green fluorescent intensity of Annexin V-FITC ( P<0.05). (2) Compared with the control group, the 2-DG group had significantly enhanced Fis1 protein fluorescent intensity ( P<0.05); the astragaloside IV+2-DG group had significantly decreased Fis1 protein fluorescent intensity compared with 2-DG group ( P<0.05); Compared with astragaloside IV+2-DG group, TPEN+astragaloside IV+2-DG group had significantly enhanced Fis1 protein fluorescence intensity ( P<0.05); compared with the control group, the TPEN+2-DG group had significantly enhanced Fis1 protein fluorescent intensity ( P<0.05). (3) TMRE fluorescence intensity in 2-DG group was significantly decreased compared with control group ( P<0.05); TMRE fluorescence intensity in astragaloside IV+2-DG group was significantly enhanced compared with 2-DG group ( P<0.05); TMRE fluorescence intensity in TPEN+astragaloside IV+2-DG group was significantly decreased compared with astragaloside IV+2-DG group (P<0.05); TMRE fluorescence intensity in TPEN+2-DG group was significantly decreased compared with the control group ( P<0.05). Conclusion:Astragaloside IV can exert neuroprotective effects through Zn 2+ inhibiting endoplasmic reticulum stress-induced apoptosis and preventing mPTP opening in PC12 cells, whose mechanism may be related to Fis1 and Bap31 expressions.

Objective:To explore the clinical characteristics of hemoglobin H (HbH) disease during pregnancy, and evaluate the impact of prepregnant anemia on maternal and fetal outcomes.Methods:Clinical data of 12 pregnant women, who were diagnosed with HbH disease by genetic testing in the Third Affiliated Hospital of Guangzhou Medical University from January 1, 2017, to December 31, 2022, were retrospectively collected. Thalassemia genotypes, general conditions before and during pregnancy, and perinatal outcomes were analyzed using descriptive analysis.Results:The 12 patients were categorized into moderate anemia (Hb<90 g/L, n=5) and mild anemia (Hb≥90 g/L and <110 g/L, n=7) groups according to the level of Hb before conception. In the moderate anemia group, with one deletional HbH disease and four non-deletional type, all had blood transfusion histories, and four developed severe anemia during pregnancy with four cases of fetal growth restriction. The mild anemia group comprised six deletional and one non-deletional genotype, with no severe anemia or transfusion requirement during pregnancy. In terms of maternal and fetal outcomes, two moderate anemia cases opted for pregnancy termination, while three delivered via cesarean section with two preterm infants having mild asphyxia and one full-term. In the mild anemia group, one case terminated due to intrauterine fetal death, and the remaining six continued the pregnancies, including two were delivered normally, one was assisted by forceps, and three through cesarean section. Among the six delivered cases, one was preterm birth, one infant was born with severe asphyxia and four mothers developed postpartum complications such as puerperal infection and postpartum hemorrhage. Conclusions:Clinical manifestations of HbH disease vary greatly. Individualized management should be offered based on the degree of anemia before pregnancy. For women with moderate anemia, close monitoring and appropriate intervention are required during pregnancy to improve both maternal and infant outcomes.

[Objective]To summarize the clinical experience of Professor MA Ruijie in the treatment of insomnia by using Hewei-acupuncture,with a view to illustrating the theoretical basis and clinical application of Hewei-acupuncture in the treatment of insomnia.[Methods]Through the clinical study with the teacher,combined with the review of Chinese medical textbooks and modern medical clinical research literature on insomnia.Professor MA's understanding of the etiology and pathogenesis of insomnia,the composition and basis of the acupuncture points of Hewei-acupuncture,and the theoretical and mechanistic study of Hewei-acupuncture for the treatment of insomnia were elaborated,and Professor MA's ideas of identification and treatment of this disease were analyzed.[Results]Professor MA believes that the sleep and wakefulness of human beings are controlled by the heart and spirit,and regulated by the Yin and Yang of Ying and Wei,which are related to the functions of internal organs,among which the malfunctioning of the spleen and stomach is closely related to the occurrence of insomnia.Professor MA's treatment of this disease is characterized by the importance of the theory of meridian and collateral disease and the Qi of the spleen and stomach in the middle-Jiao,and she believes that the foot Yangming stomach meridian Qi reversal,Ying and Wei are out of harmony;the spleen and stomach deficiency,the heart and spirit of the loss of nourishment;and elevation disorders,misdirection of the good offices are important reasons for the occurrence of insomnia.Hewei-acupuncture is a summary of Professor MA's clinical experience in treating insomnia and other neurological disorders by taking"Tianshu"(ST25),"Zusanli"(ST36),"Neiguan"(PC6)and"Gongsun"(SP4)as the main acupoints.This method relieves insomnia by regulating Qi of the spleen and stomach,strengthening the function of the spleen and stomach,and balancing Yin and Yang of the organism.[Conclusion]Hewei-acupuncture in the clinical treatment of insomnia is characterized by streamlining of acupoints,light pain,and good long-term efficacy,which is of practical significance in guiding the clinical treatment of insomnia.