Finite Element Simulation of Double-Stent Parallel Thrombectomy at Y-Shaped Bifurcation of Internal Carotid Artery / 医用生物力学

Objective To study the process of single stent and double-stent thrombectomy at the Y-shaped bifurcation of the ideal internal carotid artery by finite element simulation, analyze the stent-thrombus-vessel interaction during the thrombectomy process based on the simulation results, and provide guidance for improving the effect of stent thrombectomy at the bifurcation. Methods The CAD software was used to build the model and the finite element analysis software was used to simulate the process of single stent and double-stent thrombectomy. Results Thrombectomy was unsuccessful in single stent model and successful in double-stent model, and the maximum stress of thrombus during embolus retrieval was twice that of single stent, the maximum strain was 1.12 times that of single stent, and the maximum contact pressure on the surface of vessel was approximately twice that of single stent. Conclusions Double Solitaire stents can effectively prevent thrombus displacement at the bifurcation and successfully retrieve the thrombus, but there is a risk of fracture due to the high stress level in the middle section of the thrombus. The contact pressure of the vessel on the anterior artery side is higher during thrombectomy, and the risk of vessel damage is greater. Therefore, it is necessary to optimize the design of the stent-retriever to improve its flexibility.

Comparative Study Between Rome III and Rome Criteria in College Students With Functional Gastrointestinal Disorders / 胃肠病学

Background: The newly released Rome criteria in 2016 has a stricter and more precise definition of functional gastrointestinal disorders (FGIDs) when compared with Rome III criteria. The adjustment and improvement of diagnostic criteria by Rome criteria may affect the clinical diagnosis of FGIDs. Aims: To investigate the differences and the similarities between Rome III and Rome criteria in the diagnosis of FGIDs in college students. Methods: The FGIDs database of college students in Zhejiang Province established by our previous research team were further evaluated and analyzed by Rome criteria, and the incidence, psychological symptom score, overlapping of disease of FGIDs were calculated, and compared with Rome III criteria. Results: Of the 1 870 cases in database, 1 025 (54.81%) met Rome criteria of FGIDs; while 1 111 (59.41%) met Rome III criteria, the difference in detection rate was statistically significant (P <0.01). In Rome group, incidences of belching disorders (2.14% vs. 5.83%, P<0.01), irritable bowel syndrome (IBS) (2.78% vs. 6.90%, P<0.01), functional abdominal bloating/distension (1.28% vs. 4.12%, P<0.01) were significantly lower than those in Rome III group, while incidence of functional diarrhea was significantly higher (3.85% vs. 0.70%, P<0.01). Patients met Rome criteria showed a higher score of obsession⁃compulsion, depression and anxiety (P<0.05). Rome criteria caused 33 (25.58%) original IBS patients included in functional diarrhea, and 6 (4.65%) original IBS patients included in function constipation. The diagnosis of functional bowel disease overlapping with other FGIDs (belching disorders, functional dyspepsia) according to Rome III and Rome criteria were statistically different (P<0.01, P<0.05). Conclusions: Rome criteria has a stricter and more accurate definition of FGIDs, reflecting a more accurate psychological and clinical features, and identification of patients who really need treatment, resulting in a more efficient and feasible application in clinical practice and scientific research.

Identification of predicted epitopes of HLA-A*0201-restricted cytotoxic T lymphocytes derived from human papillomavirus type 11 E7 antigen / 中华微生物学和免疫学杂志

Objective To screen and identify the predicted epitopes of synthesized HLA-A*0201restricted CTL derived from HPVll E7 antigen.Methods Five HPVll E7 CTL epitope peptides and terramers consisting of HLA-A*0201 were selected by way of computer and synthesized by Sanquin company,including HPVllE7 7-15(TLKDIVLDL),15-23(LQPPDPVGL),47-55(PLTQHYQIL),81-89(DLLLGTLNI)and 82-90(LLLGTLNIV).These peptides binding to human peripheral blood-derived DCs were tested for their ability to activate T cells isolated from peripheral blood lymphocytes of HLA-A*0201 healthy individuals.the number of specific tetramer+CD8+T cells by flow cytometry,the level of the section of IFN-γ by ELISA,and the ability of the CTL to kill the target cells were observed.Results The immature DCs could be fully activated by all the five HPV11 E7 peptides.Peptide-loaded mature DCs were able to stimulate the epitope-specific T cells responses in vitro.An increased frequency(P<0.05)of T ceils specific for the E7 7-15 epitope compared to other epitopes of HPV11E7.The epitope-specific CTL of E7 7-15 induced by the activated DCs specifically killed HPV11E7 expressing 293 cell line,and in a ratio of 50:1,the specific cytolytic activity was the strongest than the others(P<0.05).Conclusion DCs loaded with HPV11 E7 7-15(TLKDIVLDL)peptide can induce highly effective and specific ectogenic processed epitopespecific CTL responses in vitro.This peptide may be the candidate for development of CTL based vaccine in the treatment of HPV infeetions.

Selective Silencing of Viral Gene Expression in HPV6bE6-positive Cell Line Treated with Specific siRNA / 中华皮肤科杂志

Objective To investigate the inhibition of pathogenic viral gene expression in HPV6bE6-positive cell line by specific siRNA, which might have great potential for clinical use. Methods B16 cells were transfected with recombinant plasmid pcDNA3.1(+)-GFP/HPV6bE6, and the positive cell clones were selected by fluorescence protein observation and RT-PCR. Four specific siRNAs, none of which shares homology with exons of known human genes, were designed and synthesized to target HPV6bE6 mRNA. Quantitative real-time PCR was performed to measure the inhibition rates of target gene expression by comparing HPV6bE6 mRNA concentrations before siRNA transfection with those after transfection. The inhibition rates of target gene expression with different siRNA concentrations of 0.2 nmol/L, 1 nmol/L, 10 nmol/L, 50 nmol/L, 150 nmol/L and with different treatment time at 24 h, 48 h, 72 h and 96 h after transfection were measured respectively. Results More than 90% reduction of HPV6bE6 mRNA was observed following treatment with HPV6bE6-siRNA, and HPV-negative cells were apparently unaffected by HPV6bE6-siRNA. The decrease of HPV6bE6 mRNA was maximal at 24 h after siRNA treatment and sustained for at least 4 days. The minimal level of siRNA to efficiently silence the homogeneous target gene HPV6bE6 was 1 nmol/L. Conclusion HPV6bE6-siRNA can efficiently and specifically silence target genes and may be developed as a potential therapeutic approach for HPV infection.

The Specific Immune Response Elicited by Recombinant CRT120/HPV6bE7 DNA Vaccine in Mice / 中华皮肤科杂志

Objective To investigate the cellular immune response to the recombinant DNA vaccine CRT120/HPV6bE7 in mice. Methods The recombinant encoding HPV6bE7, linked with CRT120, was constructed in pcDNA3.1 eukaryotic vector with the report gene GFP. The pcDNA3.1-GFP -HPV6bE7 was transfected into B16 cells by a lipofectamine kit. The C57BL/6 mice were vaccinated with recombinant DNA plamids. The T-cell phenotype in the peripheral blood lymphocytes of the immunized mice was measured by flow cytometry. The CTL activity of the lymphocytes from the spleens and lymph nodes, and the levels of IL-2 and IFN-? were analyzed. Results The constructed recombinant plasmid CRT120/HPV6bE7 was analyzed. Positive transfected cell clones were established and could stably express the target gene HPV6bE7. Compared with HPV6bE7, CRT120/HPV6bE7 plasmid enhanced to a greater extent CD8+ T-lymphocyte differentiation, the number of TCR?? T-lymphocytes and the levels of IL-2 and IFN-? (all P

Establishment of Murine Tumor Cell Line Expressing HPV types 6b and 11 E6/E7 Genes / 中华皮肤科杂志

Objective To construct four expression plasmids, pcDNA3.1-GFP/HPV6bE6, pcDNA3.1-GFP/HPV6bE7, pcDNA3.1-GFP/HPV11E6, pcDNA3.1-GFP/HPV11E7 and their transfected murine cell lines. Methods The Four recombinant expression plasmids comprising HPV6bE6,HPV6bE7,HPV11E6 and HPV11E7 linked with GFP, respectively, were constructed and transfected to B16 cells by lipofectamine kit. Positive clones were selected by G418 and observed by fluorescent microscopy and identified by RT-PCR. Results The four constructed recombinant plasmids were authenticated by restriction enzyme digestion and DNA sequencing. Under the fluorescent microscope, the green fluorescence could be observed in cytoplasm and nucleus of four transfected B16 cell lines. The RNA extracted from positively transfected clones resistant to G418 were analyzed by RT-PCR, which demonstrated the presence of four expected fragments. Conclusions The transfected murine cell lines B16 can express HPV6bE6,HPV6bE7,HPV11E6 and HPV11E7 gene. These transfected cell lines can be further transplanted to mice in order to investigate the biological properties and immunological mechanisms of these genes in vivo.

Effects on Immunophenotypes of Dendritic Cells Induced by Crude Extracts from Lesions of Condyloma Acuminata / 中华皮肤科杂志

Objective To investigate the effects of crude extracted proteins from lesions of condyloma acuminata on immunophenotypes of dendritic cells.Methods Plastic-adherent mononuclear cells(MNCs)were isolated from umbilical cord blood or peripheral blood and cultured in media containing cytokines(GM-CSF,IL-4and LPS).The morphology and phenotypes of these cells were analyzed by flow cytometry and microscopy in12day's culture.Cells on the fourth day were incubated with crude extracts from lesions of condyloma acuminata,foreskin proteins,and PBS,respectively,followed by phenotypic analysis after9-12days' culture.Results Expression of antigens CD1a,CD80,CD86,MHC-I,MHC-II,CD14,CD54was detected after12days' cul-ture.It was shown that MNCs could be induced to differentiate to mature dendritic cells in our culture system.After incubation with crude extracts from lesions of condyloma acuminata for another9-12days,expression of CD86and HLA-DR was increased on dendritic cells.Conclusions Compared to foreskin and PBS pulsed dendritic cells,expression of CD86and HLA-DR is upregulated on dentritic cells after pulsing with condyloma acuminata lesion proteins.The data suggest that crude extracts from lesions of condyloma acuminata might enhance antigen-pre-senting capacity of dendritic cells and strengthen activation of T lymphocytes.