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Objective:To study the morphologic features of the fusion site of proximal tibial epiphysis in normal adults and analyze its potential clinical value based on Mimics three-dimensional (3D) reconstruction.Methods:CT images of knee joint of 68 patients without obvious abnormalities of lower limbs were retrospectively analyzed in electronic database of our hospital from June 2020 to June 2021, including 41 males and 27 females. The mean age of the patients was 38.7±8.4 years (range, 25-55 years), and the mean body mass index (BMI) was 25.3±4.0 kg/m 2 (range, 18.75-41.8 kg/m 2). Mimics 3D reconstruction technique was used to reconstruct the 3D model of the proximal tibia and epiphyseal fusion site. The relationship between the surface area of epiphyseal fusion site and age and BMI was studied, and the changes of cortical thickness and density at epiphyseal fusion site were also explored. Results:The fusion site of adult epiphyseal reconstructed by Mimics 3D reconstruction is a complex wavy surface structure in 3D space. The surface area of the epiphyseal fusion site was 2,994.7±645.3 mm 2 (range, 1,704.0-4,650.0 mm 2) obtained by 3-Matic Research 12.0. The fusing area of male epiphysis was 3 269.3±533.9 mm 2 than that of female 2,577.6±578.7 mm 2, the difference was statistically significant ( t=5.06, P<0.001). However, there was no significant correlation between the epiphyseal fusion site surface area and age ( R2=0.02, P=0.268) and BMI ( R2=0.04, P=0.125). Mimics software was used to obtain the CT values of bone cortex at the epiphysis line and the distal end of the epiphysis line at 10 mm and 20 mm levels as 451.059±74.953 Hu, 1,018.412±125.732 Hu and 1,414.162±107.848 Hu, respectively. The thickness of bone cortex was 1.814±0.090 mm, 2.511±0.089 mm and 3.189±0.185 mm at 10 mm and 20 mm layers of epiphysis line and distal epiphysis line, respectively. Conclusion:In this study, Mimics 3D reconstruction technique was used to visualize the fusion site of the proximal tibial epiphysis in normal adults. The epiphyseal fusion site of adult is a undulating plate-like structure, and the cortical bone density of epiphyseal fusion site is low and thin, theoretically, it is easy to fracture under indirect violence.
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Objective:To compare the biomechanical properties of triangular supporting fixation and Gamma nail fixation for intertrochanteric fractures of the femur.Methods:The femoral CT imaging data provided by a healthy adult male volunteer aged 40 years, height 172 cm, and weight 75 kg were used to reconstruct the femur model using Mimics 21.0 software and Geomagics 2013 software. Evans type I intertrochanteric fracture models were established using UG12.0 software, and Gamma nail and triangular supporting intramedullary nail models were reconstructed to simulate intertrochanteric fracture internal fixation, respectively. In Abaqus software, two internal fixation models of Gamma nail and triangular supporting intramedullary nail in standing state are simulated, and the stress peaks of the main nail, fixation screw and bone substance were collected, also the stress peak of supporting screw of the triangular supporting intramedullary nail is obtained. Additionally, the maximum displacement of the fracture model fixed by Gamma nail and triangular supporting intramedullary nail is measured.Results:Under the load of 1 200 N, the peak stress of the two fracture internal fixation models was located in the main nail, in which the peak stress of the triangular supporting intramedullary nail was 233.73 MPa, which was 11.9% lower than that of the Gamma nail (265.21 MPa); the peak stress of the fixation screw was located in the contact area between the pressure screw and the main nail, which was 23.2% lower in triangular supporting intramedullary nail than that of the Gamma nail (138.86 MPa vs. 180.75 MPa); the peak stress of the bone model was located in the medial cortex of the femur, which was 61.67 MPa and 32.38 MPa, respectively, 47.5% lower in the triangular supporting intramedullary nail than that of the Gamma nail; the peak stress of the supporting screw in the triangular supporting intramedullary nail was 92.04 MPa. The maximum displacement of the fracture model fixed with triangular supporting intramedullary nail was 17.34 mm, which was 10.5% less than the maximum displacement of the fracture model fixed with Gamma nail (19.37 mm). Conclusion:Compared with Gamma nail, triangular supporting intramedullary nail fixation can significantly improve the stability of intertrochanteric fractures and stress distribution as well as reduce stress peak and stress concentration area, which is helpful to improve the efficacy of intertrochanteric fractures.
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Objective@#To explore the relationship between abuse experience with suicidal ideation and suicide attempts of junior middle school students, and to provide a reference for suicide prevention of junior middle school students.@*Methods@#Cluster sampling method were used to selct 10 289 junior middle school students from 25 districts and counties of Chongqing were included in the analysis of this study from July to September in 2020. And Questionnaire on Abuse in Childhood and Mental Health Scale for Middle School Students were applied to collect the data about demographic information, suicide ideation and behavior.@*Results@#The prevalence of suicidal ideation and suicide attempts among junior middle school students were 20.93% and 10.83%, respectively. Multivariate Logistic regression model found that after controlling for demographic variables and mental health, emotional abuse ( OR =2.07) and emotional neglect ( OR =2.03) showed higher correlations with suicidal ideation than the other three types of childhood abuse( OR physical neglect =1.19, OR physical abuse =1.60, OR sexual abuse =1.37, P <0.05); and sexual abuse ( OR =2.29) and physical neglect ( OR =1.87) showed higher associations with suicide attempt than the other three types of abuse( OR emotional abuse =1.63, OR emotional neglect =1.59, OR physical abuse =1.50, P <0.01).@*Conclusion@#All five types of child abuse were independent risk factors for suicidal ideation and suicide attempts, and emotional neglect and emotional abuse had a greater effect on suicidal ideation, sexual abuse and physical neglect had a greater effect on suicide attempts.
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In order to construct recombinant baculovirus carrying Schistosoma japonicum 26 ku glutathione S-transferase gene (Sj26), and observe the expression of Sj26 in mammalian cells, the Sj26 gene was amplified with plasmid pGEX-3X as template by PCR, and then recombined into T vector for sequencing. Sj26 gene was inserted into the downstream of CMV promoter of donor plasmid pFBDGC, and the recombinant donor plasmid pFBDGC-Sj26 transformed into DH10Bac, then the recombinant bacmid AcCMVSj26 was isolated and transfected into Sf9 cells. The recombinant baculovirus was harvested and final titer of vAcCMVSj26 was measured. BHK cells were transducted with recombinant baculovirus in vitro. By using Western blot, the expression of 26 ku glutathione S-transferase (GST) was detected. The results showed that after enzyme digestion and sequencing, the donor plasmid was successfully constructed. PCR confirmed that pFBDGC-Sj26 and Bacmid homologous recombination occurred in E. coli. After transfection of Sf9 cells with recombinant Bacmid, recombinant baculovirus was replicated in Sf9 cells and expressed green fluorescent protein. PCR further revealed recombinant baculovirus contained Sj26. The titer of the harvested baculovirus was 1.24 x 10(8). Western blot demonstrated that recombinant baculovirus could express 26 ku GST in BHK cells. It was concluded that Sj26 recombinant baculovirus was successfully constructed, and the 26 ku GST was expressed in mammalian cells.
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Objective To compare a potential role of dendritic cells (DCs) and macrophages in inducing protective immunity against infection with Schistosoma japonicum. Methods DCs and macrophages were pulsed in vitro with soluble egg antigen (SEA) of S. japonicum. BALB/c mice were injected three times with DCs or macrophages, either antigen-pulsed or not,and challenged with 40 ± 2 cercariae of S. japonicum per mouse. Worms were collected 42 days later by portal perfusion of the mice and egg number of liver was calculated. To evaluate whether protective immunity had been induced by preparations of DCs or macrophages, the worm burden and fertility ( eggs per female per mouse liver) were compared between the groups of mice. The antibody level against SEA was detected by ELISA. Results With respect to mice injected with untreated cells, numbers of worms and eggs per female worms were significantly reduced in the groups of mice having received pulsed DCs (26. 3% and 37.9%, respectively), or pulsed macrophages (22. 0% and 30.7%). Untreated DCs and macrophages induced no significant effects. The antibody level against SEA rose in sera of all groups of mice up to 42 days after the challenge, but most pronounced in those immunized with pulsed DCs, although this was not significantly different from other groups. Conclusion The results suggest that the protective immunity against S. japonicum might be induced by DCs to a higher extent than by macrophages after in vitro pulsing with egg antigen.
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In order to construct recombinant baculovirus carrying Schistosoma japonicum 26 ku glutathione S-transferase gene (Sj26), and observe the expression of Sj26 in mammalian cells, the Sj26 gene was amplified with plasmid pGEX-3X as template by PCR, and then recombined into Tvector for sequencing. Sj26 gene was inserted into the downstream of CMV promoter of donor plasmid pFBDGC, and the recombinant donor plasmid pFBDGC-Sj26 transformed into DH10Bac,then the recombinant bacmid AcCMVSj26 was isolated and transfected into Sf9 cells. The recombinant baculovirus was harvested and final titer of vAcCMVSj26 was measured. BHK cells were transducted with recombinant baculovirus in vitro. By using Western blot, the expression of 26 ku glutathione S-transferase (GST) was detected. The results showed that after enzyme digestion and sequencing, the donor plasmid was successfully constructed. PCR confirmed that pFBDGC-Sj26 and Bacmid homologous recombination occurred in E. coli. After transfection of Sf9 cells with recombinant Bacmid, recombinant baculovirus was replicated in Sf9 cells and expressed green fluorescent protein. PCR further revealed recombinant baculovirus contained Sj26. The titer of the harvested baculovirus was 1.24 × 108. Western blot demonstrated that recombinant baculovirus could express 26 ku GST in BHK cells. It was concluded that Sj26 recombinant baculovirus was successfully constructed, and the 26 ku GST was expressed in mammalian cells.
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Objective To study the presence of iNOS in Schistosoma japonicum and demonstrate its distribution in different stages of this schistosome. Methods Cryostat sections for adult worms and paraffin sections for eggs in the liver of infected mouse, sporocysts and cercariae in snails were prepared, immunofluorescent test was performed to detect the presence of iNOS in adult worms, sporocysts, cercariae and miracidium, the distribution of this enzyme was observed in different stages of Schistosoma japonicum. Western blotting was used to further demonstrate the immunoreactivity to iNOS in adult worms. Results The results of immunofluorescent test showed that specific yellow- green fluorescence was mainly among subtugment of adult worms. Positive staining was also distributed on the surface of miracidium and its glands. For both sporocysts and cercariae, the majority of fluorescence was on their surface. Anti-iNOS antibody could recognize an apparent specific band in Western blotting of adult worm proteins, with a of Mr 210 000. Conclusion There is an expression of iNOS-like enzyme in Schistosoma japonicum.
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Objective To study the in vitro larvicidal activity of nitric oxide (NO) to the juvenile Schistosoma japoni-cum. Methods Macrophages were induced by LPS or LPS + IFN-? to produce NO, schistosomula obtained mechanically from cercariae were added to the medium with activated macrophages, the larvicidal activity was observed within 48 h . In order to further confirm the effect of NO, an inhibitor of iNOS,L-NNA (N?-nitro-L-arginine), was used to inhibit the production of NO, larvicidal activity was measured by the same methods and the difference of dead larvae ratio was compared between the inhibited and uninhibited groups. Results LPS and LPS + IFN-? can induce macrophages effectively, with the NO production of (109.96?3.70)?mol/L and (113.50?7.38) ?mol/L respectively, accordingly the larvicidal effect reached to 91.07% ?2.92% and 96.86%?2.36% respectively. This activity can be inhibited by L-NNA. NO production and dead larvae ratio were reduced significantly in the inhibited group than in the uninhibited one. Conclusion NO produced by activated macrophages can kill schistosomula of Schistosoma japonicum.
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Objective To investigate the effect of nitric oxide on the liver fibrosis of mice infected with Schistosoma japonicum during the early stage of schistosomiasis. Methods NMRI mice were treated with AMG-an iNOS inhibitor from day 23 post infection(p.i) to the sacrificed and the livers were collected at 38 days, 45 days p.i respectively. The expression and distribution of collagen typeⅠ, Ⅲ (ColⅠand ColⅢ) in liver tissues were investigated with the Picric acid-Sirius red staining techniques and differentiated with the polarization microscopy combining with picture analysis system. Hydroxyproline concentration in liver homogenate was measured by the biochemical methods. Results At 38 day p.i, the Picric acid-Sirius red staining showed that the hyperplasia of Col Ⅰ and ColⅢ in the livers of AMG treated mice increased significantly compared with the control livers, but there was no significant difference of hydroxyproline content between the two groups. At 45 days p.i, only the hyperplasia of Col Ⅰ in AMG treated group increased significantly compared with the control livers, and moreover, content of hydroxyproline of the inhibited mice was significantly higher than that of the control mice (P
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Objective To explore the mechanism of protective immunity against Schistosoma japonicum infection induced by Sj26 gene transfected dendritic cell(DC).Methods 48 BALB/c mice were divided randomly into 4 groups with 12 each.The mice were injected through auricle for three times with Sj26 gene transfected DC(Group A),pcDNA3 transfected DC(Group B),untreated DC(Group C) and RPMI-1640(Group D) respectively,and challenged with 40?2 cercariae of S.japonicum per mouse 2 weeks after the last immunization.Sera from mice were examined for IgG antibody,IFN-? and IL-4 by ELISA.Western blot was used for detecting specific anti-Sj26 IgG antibody.The production of IFN-? and IL-4 in the supernatant of spleen cells stimulated with soluble egg antigen(SEA) and ConA was quantified by sandwich ABC-ELISA.The proliferation of spleen cells were measured with MTT method.Results IgG antibody increased significantly in the mice of group A at 2 weeks after the last immunization(absorbency A491=0.117),higher than that of group B(A491=0.061) and group C(A491=0.058)(P
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Objective To study the association of human leukocyte antigen classⅡ(HLA-Ⅱ) alleles with genetic susceptibility and resistance to advanced hepatosplenic schistosomiasis japonica. Methods The allelic types of HLA-DRB1, DPA1, DQA1 and DQB1 were detected by polymerase chain reaction with sequence-specific primers (PCR-SSP) technique in 46 patients with advanced hepatosplenic schistosomiasis, characterized with extensive liver fibrosis. Another 43 subjects with chronic schistosomiasis were used as control. The statistical significance of differences in allelic frequencies was determined by ? 2 test. Results The frequencies of HLA-DRB1*04, DPA1*0103, DQA1*0601, DQB1*0201 in advanced patients were markedly higher than those in control group, while the frequencies of HLA-DQA1*0501 and DQB1*0601 in control group were higher than those in advanced patients. Conclusion The study indicated that HLA-DRB1*04, DPA1*0103, DQA1*0601 and DQB1*0201 showing a positive, statistically significant (P
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Objective To study the expression of inducible nitric oxide synthase (iNOS) in livers of mice infected with Schistosoma japonicum. Methods The livers of NMRI mice infected with S. japonicum were collected on day 21, 28, 38, 45 post infection(p.i.), total RNA of livers were extracted and kinetics of the mRNA expression of iNOS were detected by RT-PCR, the protein expression of iNOS was then confirmed by Western blotting and the distribution of iNOS in the infected liver was determined by immunohistochemical methods. Results The mRNA expression of iNOS was not detectable in the uninfected liver, iNOS mRNA expression was detected on day 21 p.i, the expression increased on day 28 p.i and peaked on day 38 p.i, then decreased slightly on day 45 p.i. Western blotting showed an iNOS expression in the livers only on day 38, 45 p.i. IFA test showed that the expression of iNOS was maily distributed in the granuloma of the livers. Conclusion S. japonicum infection can induce the expression of iNOS in a time-dependent manner in the liver of the host,and eggs may be the main factor in inducing the expression.
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Objective To explore the presence of biliverdin reductase (BR) activity in adult worms of Schistosoma japonicum in vitro. Methods The soluble fraction was isolated from homogenates of adult worms of S. japonicum, and incubated with biliverdin under different pH conditions and buffers. The time dependency of this enzymatic reaction was also detected. ResultsThe soluble fraction of the homogenates of adult worms could degrade biliverdin in vitro, the BR activity was 43.30 nmol/(mg?min), with its optimal condition at pH 8.7. The rate of reaction peaked at 15 min of incubation for the BR activity. Conclusion The presence of BR activity in adult worms of S. japonicum was firstly demonstrated.
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Objective To construct and express recombinant plasmid containing the structural gene encoding {Mr 31 000} antigen of Trichinella spiralis muscle larvae. MethodsThe target gene TspE1 was amplified by RT_PCR, cloned into pUC18 vector, and sub_cloned into the eukaryotic expression vector pcDNA3. BALB/c mice were immunized with the purified recombinant plasmid pcDNA3_TspE1 by gene_gun bombardment. The expression of recombinant plasmid in the skin tissue was observed by HE staining and immunohistochemical staining. Results and Conclusion The recombinant plasmid pcDNA3_TspE1 was successfully constructed and expressed in the BALB/c mice.
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Objective To detect the infection of Schistosoma japonicum in mice with a novel test based on agglutination of hybridoma cells and to study the mechanism of the hybridoma cells agglutination. Methods The procedure was developed with a murine cell line H226 producing a monoclonal antibody specific to schistosome 31/32 kDa antigen and sera collected from mice infected with different numbers (10,30,50) of S. japonicum cercariae in different period. Immunofluorescent test was carried out with the hybridoma cells and schistosome-infected sera. Results The circulating antigen was detected by the test as early as 2 weeks after a heavy infection and all mice showed positive results in the test by 5 weeks after infection. The titers of antigen rose along with the lime post infection, and the tilers of sera from heavy infection were statistically higher than that from the mice receiving a lower number of cercariae. Specific yellowish green fluorescence appeared on the membrane of the hybridoma cells; no signal was detected inside. Conclusion Hybridoma cell agglutination test (HCAT) may become useful to diagnose schistosomiasis.
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Objective To explore the activity of heme oxygenase and immunolocate the enzyme in the adult worms of Schistosoma japonicum .Methods Microsomal protein was isolated from the homogenate of adult S^japonicum , heme degradation and effect of different pH conditions and buffers on degrading reaction were investigated by incubating microsomal protein with hemin. The slices of whole worm and cells of S^japonicum were prepared, distribution of HO in schistosome was studied by immunofluorescent and alkaline phosphatase(AP) \|immunocytochemical assays.Results Microsomal protein of adult worms can degrade the heme in vitro , the activity being 56^7 nmol bilirubin/(mg?min).The optimal pH was 8^7. Immunofluorescent and AP\|immunocytochemical assays revealed that the HO distributed dispersively in the worm, and located in cytoplasm.Conclusion The presence of HO was firstly proved in S^japonicum .
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The common antigen component between B. malayi and O. armillata was demonstra-ted by cross immunoelectrophoresis(XIE) with antigens from these 2 species of filariae against the sera from patients infected with B. malayt. When the two antigens were subjected to SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and enzyme linked im-munoblotting (ELIB), the component of approximately 43 KD was identified to be the common antigen component.
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0.05). In the control test against tne sera of normal persons, the false positive rate of the two antigens was both 7.7%(4/52).It is suggested that O. armillata adult worm may be used as a heteroantigen for diagnosis of filariasis.