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Objective@#To investigate the correlation between CD47 and cervical lesions.@*Methods@#The expression and overall survival of CD47 in cervical cancer was analyzed using TCGA and TCGA Target GTEx databases. Based on GEO databases , the expression of CD47 was analyzed between cervical sample infected by HPV 16/18 and nonHPV. The differences of CD47 expression in the degrees of cervical lesions infected by HPV 16/18 were compared.The patholgical sections of colposcopy biopsy came from 320 patients who were admitted cervical cancer screening ,including HPV typing and E6/E7 mRNA detection. Immunohistochemistry were used to determine CD47 expression , and histochemistry score(H⁃score) was used to quantify the levels of CD47. The correlation among HPV 16/18 , pathological grades and CD47 was analyzed.@*Results@#Bioinformatics tool analysis and immunohistochemistry showed that CD47 was up⁃regulated in HSIL and CC , associated with the prognosis of CC. The expression levels of CD47 in cervical tissues increased significantly after HPV 16/18 infection ( t = 2. 494 , P < 0. 05 ) , and in HPV 16/18 infected tissues , with the increase of pathological grades. The expression levels of CD47 significantly increased(F = 4. 351 , P < 0. 05 ; rs = 0. 278 , P < 0. 001) . The expression of CD47 in the E6/E7 mRNA( + ) was higher than that in the E6/E7 mRNA( - ) ( t = 5. 710 , P < 0. 000 1) , and the copies of E6/E7 were related to the degree of cervical lesions , and were positively correlated with the expression level of CD47 ( F = 15. 557 , P <0. 000 1 ; rs = 0. 649 , P < 0. 000 1) ( rs = 0. 73 , P < 0. 000 1) .@*Conclusion@#CD47 is overexpressed in HSIL and CC after HPV 16/18 infection. The expression levels of CD47 increases with the degree of cervical lesions , and is correlated with E6/E7 mRNA. CD47 may be a potential target for cervical disease progression and therapy.
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Objective To explore the correlation between myeloperoxidase (MPO) genetic variation and coal-burning endemic fluorosis, and to understand the influence of integrated intervention including stove changes and health education on people’s health in the area. Methods In 2007, coal-burning endemic fluorosis disease areas were selected in Bijie City, Guizhou Province. No stove changes in Yachi Town, 150 patients with dental fluorosis were selected as fluorosis non-intervention group, and the intervention group was 150 patients in Changchun Town where the stoves were changed 2 years ago. The population in control group was selected in an area with non-endemic fluorosis in Changshun County. The mRNA expressions of MPO in leukoxytes were detected by real-time PCR. HepG2 cells were cultured in vitro and divided into four groups: pGL3-A group, pGL3-G group, pGL3-Control group and pGL3-Basic group. pGL3-A and pGL3-G were recombinant plasmid, while pGL3-Basic as a blank control and pGL3-Control as a positive one. The internal reference plasmid pRL-TK co-transfected the HepG2 cells with pGL3-G, pGL3-A, pGL3-Basic and pGL3-Control, respectively. The influence of sudden change of MPO gene promoter on the gene transfection activity was evaluated by a dual luciferasereporter gene system. Results The expression level of MPO mRNA in peripheral blood leukocytes in non-intervention group(0.054 ± 0 . 003 ) were higher than control and intervention groups (0.019 ± 0.004,0.019 ± 0.003, all P0.05). After the MPO-463G/A locus genetic variation occured, the luciferase reporter gene expression level of the recombinant plasmid pGL3-G(0.753 4 ± 0.086 6) was higher than that of the pGL3-A(0.490 0 ± 0.022 3, P < 0.05). Conclusions The study on MPO gene promoter-463G/A locus has prompted that MPO gene allele may be a protective factor to coal-burning fluorosis. The integrated interventions have a role in the prevention and treatment of endemic fluorosis.
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Objective To understand the dietary intake of pregnant women, and try to give a reasonable sug-gestion to promote fetal development. Methods 400 pregnant women in Fushan city were enrolled in this study and their dietary intake were investigated, then the software of Analysis of pregnancy nutrition guidance system was used to make nutrition analysis. Results The ratio of the energy in each pregnancy provided by dietary protein, carbohy-drate and fat were relatively appropriate. But the intake of dietary calcium, ferrum, zinc, vitamin B1 and vitamin B2 in early pregnancy were very low, the intake were respectively 764.51 mg, 26.94 mg, 15.71 mg, 1.12 g, 1.13 g. Each pregnancy the ratio of high-quallty protein in total protein intake were more than 50%, an average protein RNI% was 48.50%. Conclusion The dietary style of pregnant women is relatively reasonable. But they should strengthened nutritional guide to intake more calciuro,ferrum, zinc,vitamin B1 and vitamin B2.
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To investigate the effect of semi-joint prosthesis replacement in treating osteosarcoma around the knee joint,15 children with osteosarcoma around the knee joint underwent semi-joint prostheses replacement and removal of the section with tumor from October 1998 to July 2006.Imageology showed that tumor of 9 children was located at the distal femur,and 6 cases at the proximal tibia.The pathologic changes involved the upper segment of the tibia 9-11 cm in length.All the children had been diagnosed with osteosarcoma by pathological examination,which was graded as stage Ⅱ a according to Enneking staging system.All the patients underwent neoadjuvant chemotherapy preoperatively for 1-2 courses.The size of prosthesis was determined based on preoperative X-ray,CT and MRI.The length of the prosthesis should be 1.0-2.0 cm longer than that of the excised bone.Patellar and collateral ligaments were reconstructed in the artificial semi-knee joint prosthesis.All the 15 patients had a successful semi-joint prosthesis replacement.During the follow-up of 2 to 6 years,body height of the children increases by 4-6 cm,and the affected limbs shortened 1-3 cm.All children could live and study normally.There were 7 excellent,5 good,2 fine and 1 poor according to Enneking evaluation system.The semi-joint prosthesis replacement in the limb salvage surgery for child osteosarcoma around the knee joint can reconstruct knee joint function,remain normal osteoepiphysis,minimize affected limb crispation after operation,and create the condition for total knee replacement in adults.
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BACKGROUND:There is a linker between Hsp65 and Esat6 coding a segment of water repellent polypeptide.It is very gentle and easy to be folded,which is profitable to translate the keno-folding correctly between the two proteins and to make the space structure of fusional proteins consistent with those two native ones.Then,a correct structure is formed and the immunogenicity of the fusional proteins is improved.OBJECTIVE:This study was to clone the fusional genes Hsp65-Esat6 from mycobacterium tuberculosis(MTB)H37Rv, then to construct into a eukaryotic expressing vector which contains an enhanced green fluorescent protein(EGFP) reporter gene,and finally to identify the expression of Hsp65 protein and Esat6 protein by IHC methods.DESIGN:Single sample experiment.SETTING:Department of Microbiology ＆ Immunology,Medical College of Jinan University.MATERIALS:Plasmid pVAE was donated by Professor Cao from South China University of Technology.MTB H37Rv,E.coil DH5 α,expressing plasmid pEGFP-C1 and Hela cells were donated by Doctor Hu Ping.METHODS:The experiment was conducted at the Department of Microbiology ＆ Immunology and the Medical Experimental Center of Jinan University between September 2005 and June 2006.The whole-genome was extracted from MTB H37Rv by molecular cloning technique,and used it as template to amplify Hsp65 (no terminator) gene by polyrnerase chain reaction (PCR),to recombine it with pEGFP-C1 vector after purification to construct pEGHsp65(no terminator)recombination vector.pVAE vector was used as template to amplify Linker-Esat6 gene(with terminator)by PCR, and then to recombine it with pEGHSP65(no terminator)to construct an eukaryotic expressing vector pEGHsp65-Esat6 with the fusional genes Hsp65-Esat6 inside.Finally,genes Hsp65-Esat was checked by molecule biology methods such as PCR, restriction endonuclease and DNA sequencing.Hela cells were transfected with pEGHsp65-Esat6 and the expression of EGFP and the efficiency of transfection were observed.The expressions of Hsp65 protein and Esat6 protein were detected by IHC methods.MAIN OUTCOME MEASURES:①The size of pEGHsp65-Esat6.②The DNA sequencing result of the pEGFP-C1.③The expression of EGFP in the transfected Hela cells.④The IHC results of Hsp65 protein and Esat6 protein in Hela cells.RESULTS:①The size of pEGFP-C1 was 4.7 kb,that of pEGHsp65 was 6.4 kb,and that of pEGHsp65-Esat6 was 6.7 kb.There were differences between their speeds In agarose electrophoresis.②The results showed that it was the same as reported Hsp65 sequence and Esat6 sequence of MTB H37Rv.③After transfecting pEGHsp65-Esat6 for 24 hours,EGFP was found in 30% of Hela through Laser scanning confocal microscope. But there was no EGFP in non-transfected Hela.④Hsp65 protein and Esat6 protein with biological activities were detected in transfected Hela cells by IHC methods.CONCLUSION: Using Hsp65 and Esat6 as immunogens,we have successfully cloned and constructed a eukaryotic expressing vector which contain fusionaI genes Hsp65-Esat6 of MTB H37Rv and EGFP.