Objective To construcr a phage display library of human single-chain Fv antibodies from peripheral blood lymphocytes of people with positive serum antibody against hepatitis B virus core protein. Methods The heavy chain and light chain variable region gene of human immunoglobulin were amplified individually by RT-PCR from peripheral blood Lymphocytes mRNA of patients with positive serum hepatitis B virus core protein antibody and randomly combined through a DNA linker encoding the peptide (Gly 4Ser) 3 to construct single-chain variable fragments (ScFv)gene. In the presence of helper phage M13KO7, the ScFv were displayed on the surface of recombinant phages. Results Repertoire antibody library was obtained with 10 6 clonies resistant to ampicillin and kanamycin. Conclusions A typical phage display library of human single-chain Fv antibodies has been constructed Using RT-polymerase chain reaction (RT-PCR) and phage display technique, human single-chain Fv antibodies can be screened from this library.

The training of teaching ability is an important context of master course.Pathology Department of Xiang Ya School of Medicine take experimental teaching practice as the important way to improve their pathological experimental teaching skill.In this paper we introduced our experience.

Tissue engineering composites for bone defect treatment is currently a hot spot of the research field.Mechanical loading on tissue engineering composites is important to bone formation and related research has attracted more and more attention.In the field of tissue engineering research,the establishment of a standardized animal model is the basis of experimental research for exploring the effects of different mechanical loading on in situ repair of bone defect and its action mechanism.This paper reviews the approaches used to establish experimental model of bone defect under mechanical loading such as animal selection,defect preparation,fixation and loading mode in order to provide a reference for related research.

AIM: To Screen and identify human single-chain variable fragment (ScFv) specific to hepatitis B virus core protein and determine its gene sequence. METHODS: The recombinant phages were panned by HBcAg coated in a 96-pore plate and 48 clones were identified specific to HBc after three rounds of panning. The specificity of ScFv from the positive clone was determined by ELISA. Then, the soluble ScFv was expressed in E.coli. HB2151 and secreted in the supernatant. Subsequently, SDS-PAGE and dot blot were performed to identify the ScFv in the supernatant and cell lysate. The gene of ScFv specific to hepatitis B virus core protein was sequenced. RESULTS: The ScFv screened from phage antibodies has a specific combination character with hepatitis B virus core antigen. Soluble ScFv was confirmed to express in E.coli. HB2151 and secrete in the supernatant. The sequence of ScFv gene conformed to that of heavy chain and kappa chain of human immunoglubulin. CONCLUSION: Human ScFv specific to hepatitis B virus core protein has been identified by means of the phage display technology, and its gene sequence has been determined.

AIM: To observe the change of nitric oxide (NO) generation system in the vascular adventitia, media and intima in septic shock rats. METHODS: The septic shock model was made in rats by caecal ligation and puncture. The intima, media and adventitia of the rat aorta were separated. NO production (NO - 2) , nitric oxide synthase(NOS) activity and L-arginine (L-Arg) transport were measured, separately. Inducible NOS (iNOS) distribution was detected by immunohistochemistry. RESULTS: Both in early and late stage of septic shock, NO - 2 from the intima was decreased by 66.1% and 78.9%( P

Objective Within human hepatoma cell lines,we aimed to investigate the effects of the down-regulation by RNAi on different fragments of osteopontin (OPN) in order to discover more effective and accurate sites for OPN.Methods Specific small interfering RNA of OPN (OPNi-1) were synthesized and transfected into human hepatoma cell line (HEP-G2).Fluorescent quantitative PCR and immunohistochemical methods were used to test*the OPN expression levels of mRNA and protein before and after RNAi.Results After transfection,the △CT value of the A fragment was greater than B and C fragments of OPN mRNA in HEP-G2.Before RNAi was added to HEP-G2 cells,the three fragments A,B,C had OPN mRNA CT values of 8.31±1.58,8.78±1.49,8.25±1.51 respectively.Once the RNAi were added,the CT values were measured 48h after for the fragments A,B,and C which were 12.14±1.43,10.22±1.97,10.48±1.88 (P＜0.05) respectively.The immunohis tochemical values of A,B,C were down from 6.44±1.67,5.43±2.05,5.45±2.52 to 2.84±1.52,4.43± 1.65,3.95± 1.43 respectively after interference.Conclusions RNAi can inhibit the expression of OPN gene selectively.siRNA targets different segments of OPN,which may have more effects on invasion and metastasis of liver cancer for a more important significance in science and health economics.

AIM: To construct an eukaryotic expression vector of human single-chain variable fragment ~against hepatitis B virus core protein (anti-HBc ScFv) and detect its expression in HepG2 cells. METHODS: Anti-HBc ScFv genes were amplified from the plasmids abstracted from positive clone and inserted into pEGFP-c1 vector that contained green fluorescent protein gene. The recombinant plasmids were transfected into HepG2 cells, and resistant clones were obtained by G418 selection. The expression of the gene of fusion protein was determined by fluorescent invert microscope and ELISA. RESULTS: Recombinant plasmids were successfully constructed. The plasmid transfected HepG2 cells were obtained by G418 selection. Specific fluorescence was observed in HepG2 cells 48 hours after transfection. ELISA analysis confirmed the expression of anti-HBc ScFv in the cells. CONCLUSION: The construction of human anti-HBc ScFv eukaryotic expression vector and its expression in HepG2 cells lay the foundation for advanced research of intracellular anti-HBc ScFv.

Objective To propose a novel regional injury classification for open pelvic fractures and characterize its application in clinical treatment.Methods The clinical data of 67 open pelvic fractures were retrospectively analyzed which had been treated from January 2001 through December 2017 at Department of Orthopedics,Affiliated Hospital to Chengdu University.They were 48 males and 19 females,from 12 to 68 years of age (average,34 years).According to the Tile classification,23 cases were type A,19 Type B and 25 Type C.Their main injuries were located at pubic perineum in 29 cases,at ilioinguinal region in 20,at sacroiliac region in 7,at ilioinguinal-perineum in 5 and at sacroiliac-perineum in 6.Our novel regional injury classification was proposed according to the assessment of soft tissue injuries at the Faringer regions.The mortalities and complications were compared between types of our regional injury classification to find associations between them.Results Despite active treatment 28 patients died,giving a mortality of 41.8％ (28/67).Our regional injury classification grouped the open pelvic fractures into 4 types:perineal type (type Ⅰ),ilioinguinal type (type Ⅱ),sacroiliac type (type Ⅲ) and composite type (type Ⅳ).The mortalities for the 4 types were respectively 20.7％ (6/29),45.0％ (9/20),71.4％ (5/7) and72.7％ (8/11).The mortality of open pelvic fractures was positively correlated with our classification (y =0.620,P =0.001).Type Ⅰ mainly included cystourethral injury,anorectal injury and infection,type Ⅱ and Ⅲ injury to iliac vessels and Type Ⅳ injury to iliac vessels and infection.The incidences of complications for type Ⅲ (100.0％,7/7) and type Ⅳ (100.0％,11/11) were significantly higher than those for type Ⅰ (82.8％,24/29) and type Ⅱ (85.0％,17/20) (P ＜ 0.05).Conclusions Our regional injury classification gives clear criteria.The mortality increases gradually with the 4 types.The incidence of complications for each type varies and thus leads to different therapies.

CT three-dimensional reconstruction has been widely used in the diagnosis and classification of fractures. Fracture mapping based on the CT three-dimensional reconstruction can visually display the morphology of fracture lines by drawing a fracture model, showing the starting and ending points, progress and area of the fracture lines. It provides a new method for diagnosis and classification of fractures, operative plan and design of implants for fractures, statistic analyses of predominant fracture sites, and developing standardized fracture models. This review gives a brief introduction of the research progress at home and abroad in fracture mapping for scapular fracture, pilon fracture, ulnar coronoid fracture, tibial plateau fracture, radial head fracture, intertrochanteric lateral wall fracture and acetabular quadrilateral surface fracture, summarizes the likely sites in the above fracture models and clinical application of fracture mapping in fracture classification, and discusses the prospects and problems of this technique in clinical application.

Background and Objectives: Bronchopulmonary dysplasia (BPD) has major effects in premature infants. Although previous literature has indicated that mesenchymal stem cells (MSCs) can alleviate lung pathology in BPD newborns and improve the survival rate, few research have been done investigating significantly differentially expressed genes in the lungs before and after MSCs therapy. The aim of this study is to identify differentially expressed genes in lung tissues before and after hAD-MSC treatment. Methods: and Results: Human amnion-derived MSCs (hAD-MSCs) were cultured and met the MSCs criteria for cell phenotype and multidirectional differentiation. Then we confirmed the size of hAD-MSCs-EXOs and their expressed markers. An intratracheal drip of living cells showed the strongest effect on NHLI compared to cellular secretions or exosomes, both in terms of ameliorating pulmonary edema and reducing inflammatory cell infiltration. Through gene chip hybridization, PCR, and western blotting, acylaminoacyl-peptide hydrolase (APEH) expression was found to be significantly decreased under hyperoxia, and significantly increased after hAD-MSC treatment. Conclusions The intratracheal transplantation of hAD-MSCs ameliorated NHLI in neonatal rats through APEH.