Objective:To explore the value of fusion imaging in the treatment of complex aortic pathology.Methods:A retrospective analysis was conducted of 29 patients with complex aortic pathology who underwent treatment with endovascular aortic repair using fusion imaging (FI+ ) technique or without FI (FI-) between June 2015 and June 2021. The perioperative outcomes and morbidity of the FI was assessed and the early results of follow up were evaluated.Results:The mean age of patients was (70.3±7.3) years old, and 24 (82.8%) males. Technical success was 96.5% (28/29). The FI+ group patients had lower procedure time[FI+ , (209±53) min vs. FI-, (306±24)min, P=0.005]and ionic contrast medium[(169±23)ml vs. (201±20)ml, P=0.040]. Contrast-induced acute kidney injury (CI-AKI)[3.4%(FI+ 0 vs. FI-6.7%, P=0.33)], and operation-related reintervention[6.9%(FI+ 0 vs. FI-13.3%, P=0.16)] were similar. There were no significant differences in blood loss, fluoroscopy time. Conclusion:FI technique improves the accuracy during positioning in complex endovascular aortic repair, could reduce aortic related reintervention rate, operation time and contrast dose. Further studies and development are needed to obtain optimal image quality and higher precision.

Objective: To investigate the features of chest CT imaging and dynamic changes of severe coronavirus disease 2019 (COVID-19). Methods: The clinical and computed tomography (CT) data of 17 patients diagnosed with severe COVID-19 admitted to Chongqing Public Health Medical Center from January 24 to February 6, 2020 were collected. The first chest CT manifestations and the dynamic changes of imaging during treatment were retrospectively analyzed. Results: The first chest CT manifestations of the 17 patients showed that 16 cases presented with peripheral and subpleural distributions, and 2 cases presented with 3 lobes involved, one case with 4 lobes involved and 14 cases with 5 lobes involved, and 17 cases presented with ground-glass opacities, ten cases with consolidation, seven cases with subpleural line, nine cases with air bronchogram, 3 cases with thickened lobular septum, two cases with bronchiectasis, two cases with pleural effusion, two cases with lymphadenopathy with the short diameter of 1.0-1.2cm. Among 16 patients who underwent repeated CT examination, the lesions of 8 patients showed continuous improvement, and those of the other 8 patients showed fluctuating changes. Conclusions The CT findings of severe COVID-19 patients are mainly ground-glass opacities and consolidation, with the peripheral distribution. The range of lesions is wide, with 5-lobe involvement mostly. Lymphadenopathy or pleural effusion is rare. Chest CT is useful for the evaluation for the therapeutic effects.

Objective:To investigate the features of chest computed tomography (CT) imaging and dynamic changes of severe corona virus disease 2019 (COVID-19).Methods:The clinical and CT data of 17 patients diagnosed with severe COVID-19 admitted to Chongqing Public Health Medical Center from January 24 to February 6, 2020 were collected. The first chest CT manifestations and the dynamic changes of imaging during treatment were retrospectively analyzed.Results:The first chest CT manifestations of the 17 patients showed that 16 cases presented with peripheral and subpleural distributions, and two cases presented with three lobes involved, one case with four lobes involved and 14 cases with five lobes involved, and 17 cases presented with ground-glass opacities, ten cases with consolidation, seven cases with subpleural line, nine cases with air bronchogram, three cases with thickened lobular septum, two cases with bronchiectasis, two cases with pleural effusion, three cases with lymphadenopathy with the short diameter of 1.0-1.2 cm.Among 16 patients who underwent repeated CT examination, the lesions of eight patients showed continuous improvement, and those of the other eight patients showed fluctuating changes.Conclusions:The CT findings of severe COVID-19 patients are mainly ground-glass opacities and consolidation, with the peripheral distribution. The range of lesions is wide, with five-lobe involvement mostly. Lymphadenopathy or pleural effusion is rare. Pynamic monitoring chest CT is useful for the evaluation for the therapeutic effects.

Objective To develop a quantitative immunohistochemistry assay for duck hepatitis B virus core antigen (DHB-cAg)in duck liver tissue.Methods By comparison with no repair antigen and repair antigen with high pressure,microwave and trypsin,the best solution of antigen retrieval was determined.By optimizing the parameter of image acquisition and de-ducting blank area,mean density of yellow areas was calculated using Image-Pro Plus 6.0 software.Using the assay devel-oped to determine the level of DHBcAg in liver tissue from duck infected by DHBV,anti-DHBV activity of DHBcMAb-TAT PTD conj ugate was examined.Results SABC method with no repair antigen was selected,which was better than other methods.DHBcAg expression in duck liver tissue could be objectively and accurately quantified by setting Image-Pro Plus 6.0 software parameters and calculating mean density of yellow areas.By comparison with the differences between mean densityat baseline of treatment and end of treatment,it was showed that DHBcMAb-TATPTD conjugate treatment dose-de-pendently reduced the levels of DHBcAg in liver tissue,which show that the assay developed could effectively evaluate the anti-DHBV activity of agent.Conclusion The immunohistochemistry assay developed in this study can objectively and accu-rately evaluate the level of DHBcAg in duck liver tissue.

OBJECTIVE: To construct a recombinant prokaryoticexpression plasmid pET/ c-Aβ(15)-c, and evaluate the immunogenicity of its encoded fusion protein as expressed in E.coli. METHODS: The gene fragment HBc88-144 was amplified by PCR and subcloned to pUC19. The synthetic, double-strand Aβ(1-15) gene was inserted downstream of HBc1-71 in pGEMEX/c1-71. After restriction enzyme digestion, c1-71- Aβ(15) was spliced to HBc88-144, yielding the recombinant gene c-Aβ(15)-c; that gene was subcloned into pET-28a(+). The fusion protein (CA15C) expressed in the transformed E.coli BL21 was induced with isopropyl β-D-1-thiogalactopyranoside (IPTG) and analyzed by SDS-PAGE. The virus-like particle (VLP) formed by fusion protein CA15C was observed with transmission electric microscope (TEM). Four Kunming (KM) mice were given intraperitoneal injections of CA15C, and the anti-Aβ antibody elicited was detected by indirect ELISA. RESULTS: The sequence of the recombinant gene was confirmed by restriction enzyme digestion and DNA sequencing. After IPTG induction, the fusion protein was expressed, mainly in the sediment from the bacterial lysate. The expression level was 40% of total protein in the sediment. The CA15C could form VLP. After 5 rounds of immunization, the titer of anti-Aβ antibody in the sera of KM mice reached 1:10000, while the anti-HBc antibody was undetectable. CONCLUSION Recombinant c-Aβ(15)-c gene can be expressed in E.coli. The expressed protein can form VLPs and has a strong immunogenicity.
Alzheimer Disease ; prevention & control ; Amyloid beta-Peptides ; genetics ; Animals ; Base Sequence ; Genetic Vectors ; genetics ; Hepatitis B Core Antigens ; genetics ; immunology ; Humans ; Mice ; Molecular Sequence Data ; Peptide Fragments ; genetics ; Recombinant Fusion Proteins ; genetics ; immunology ; metabolism ; Vaccines, Virus-Like Particle ; genetics ; immunology ; metabolism

ObjectiveLoss-of-function mutation of p53,a tumor suppressor gene,is an important mechanism for the development of human cancers.In this study we tried to transfect p53N15-based fusion peptide into H1299,a lung cancer cell line,and evaluate the anti-tumor effects of the fusion peptide.MethodsAdeno-associated virus ( AAV) vectors were used for transfecting p53N15 fusion peptide into p53-null lung adenocarcinoma H1299 cells.The anti-replication effects of p53N15 fusion peptide were evaluated with inverted microscopy,MTT test for cell viability and flow cytometry.ResultsFusion peptides in H1299 cells was highly expressed and had detectable suppressive effects on cell proliferation.A large amount of dead cells were seen under microscope after the transfection of recombinant viruses for 72 hours.Cells activity was reduced significantly in the virus-transfecting groups as demonstrated by MTT test.The flow cytometry showed that a large number of dead cells were present in the virus-transfecting groups.ConclusionThe growth of H1299 lung adenocarcinoma cells could be inhibited in vitro by being transfected with p53N15 fusion peptide,which may be a potential gene therapy alone or as an adjuvant option in the treatment of lung cancer.

Objective It was reported that p53 apoptotic peptide (N37) could inhibit p73 gene through being bound with iASPP, which could induce tumor cell apoptosis. To further explore the function of N37, we constructed the cloning plasmid of DNA fragment encoding p53 (N37) apoptotic peptide by using DNA synthesis and molecular biology methods. Methods According to human p53 sequence from the GenBank database, the primer of p53(N37) gene was designed using Primer V7.0 software. The DNA fragment encoding p53 (N37) apoptotic peptide was amplified by using self-complementation polymerase chain reaction (PCR) method and cloned into the pGEM-T Easy vector. The constructed plasmid was confirmed by endonuclease analysis and sequencing. Results The insertion of objective DNA fragment was confirmed by plasmid DNA enzyme spectrum analysis, p53 (N37) gene was successfully synthesized chemically in vitro. The sequencing result of positive clone was completely identical to the human p53(N37) sequence in GenBank using BLAST software (http://www. ncbi. him. nih. gov/cgi-bin /BLASTn). Conclusion The cloning of DNA fragment encoding p53(N37) apoptotic peptide was constructed by using DNA synthesis and pGEM-T Easy cloning methods. With the constructed plasmid, we could further investigate the function of N37 peptide.

OBJECTIVE: To construct an universal recombinate adeno-associated virus (AAV), rAAV-NT4-ADNF-9, and to detect the ability of transfection of the rAAV vector into cochlea in vitro. METHOD: pSSVHG-CMV-ADNF-9 plasmid was introduced into 293 cell by method of Ca3 (PO4)2 using three plasmids of pSSHG-CMV-NT4-ADNF-9, pFG140 and pAAV/Ad. The recombinate adeno-associated virus (rAAV) was harvested, and the titrations of the rAAV concentrated was detected by dot-blot test. Isolate and culture the cochlear hair cell of SD rats newly born in vitro. The rAAV-NT4-ADNF-9 was added to the medium while plating. Cochlear were collected 24 h after cultivation for RT-PCR to detect the transfection of rAAV-NT4-ADNF-9. RESULT: The titration of rAAV stock produced 2 x 10(16) total particles/L, which showed that rAAV-NT4-ADNF-9 was constructed successfully. The cochlear hair cell of SD rats newly born was isolated and cultured in vitro successfully. It certified that rAAV-NT4-ADNF-9 was able to transfect into cochlear and express secretory NT4-ADNF-9 peptide by RT-PCR. CONCLUSION The rAAV vector constructed in this paper, rAAV-NT4-ADNF-9, can transfer into cochlear cultured in vitro, which layed a foundation of further research for gene therapy.
Animals ; Cells, Cultured ; Cochlea ; cytology ; Dependovirus ; genetics ; Genetic Vectors ; Hair Cells, Auditory ; cytology ; Nerve Tissue Proteins ; genetics ; Rats ; Rats, Sprague-Dawley ; Transfection

Objective To construct NT4-GFP-Ant fusional reporting gene and the vector of NT4-GFP-Ant recombinant adeno-associated virus(AAV).Methods The GFP gene was cloned by using PCR and T-vector cloning method.The positive clone was identified by the restriction enzymes,and then the cloned amplified fragments were sequenced and analyzed.The resulting gene of GFP and Ant,PBV220/NT4 were connected by DNA ligase,and thus PBV220/NT4-GFP-Ant was constructed,then the NT4-GFP-Ant fragment was gained and identified by the restriction enzymes.The resulting gene of NT4-GFP-Ant fragment was inserted into the EcoRⅠ-BamHⅠsite of vector plasmid pSSHG to construct the vector of NT4-GFP-Ant recombinant AAV.Results A 730 bp fragment of DNA was gained when T-easy/GFP was cut by the restriction enzyme EcoRⅠ.The cloned GFP gene was coincident with the sequence in GenBank.A 1 000 bp fragment of DNA was gained when pBV220/NT4-GFP-Ant was cut by the restriction enzymes BamHⅠ/EcoRⅠ.A 1 000 bp fragment of DNA was gained when PSSHG/NT4-GFP-Ant was cut by the restriction enzymes BamHⅠ/EcoRⅠ.Conclusion GFP gene is cloned successfully,NT4-GFP-Ant gene and PSSHG/NT4-GFP-Ant recombinant AAV vector are constructed successfully.

Brassinosteroid (BR) and gibberellin (GA) are two groups of plant growth regulators essential for normal plant growth and development. To gain insight into the molecular mechanism by which BR and GA regulate the growth and development of plants, especially the monocot plant rice, it is necessary to identify and analyze more genes and proteins that are regulated by them. With the availability of draft sequences of two major types, japonica and indica rice, it has become possible to analyze expression changes of genes and proteins at genome scale. In this review, we summarize rice functional genomic research by using microarray and proteomic approaches and our recent research results focusing on the comparison of cDNA microarray and proteomic analyses of BR- and GA-regulated gene and protein expression in rice. We believe our findings have important implications for understanding the mechanism by which BR and GA regulate the growth and development of rice.
Gene Expression Profiling ; Gene Expression Regulation, Plant ; physiology ; Gibberellins ; metabolism ; Oligonucleotide Array Sequence Analysis ; Oryza ; genetics ; physiology ; Proteomics