Objective To prepare the Env-pseudotyped subtype B HIV-1 with enhanced green fluorescent protein ( EG-FP) gene,explore HIV-1 infection mechanisms and develop feasible methods of identification .Methods The Env-pseudo-typed viruses were packaged in HEK293T cells by cotransfection, and the reporter gene and P24 protein were detected by PCR, Western blot and ELISA .Reporter gene amplification , viral titration assay and a single round of infection assay were performed after the env-pseudotyped viruses infected HIV-1 permissive cell .Results and Conclusion A generation and identification method of the pseudotyped HIV-1 was established . The Env-pseudotyped subtype B HIV-1 has been prepared, which is able to infect SupT1 and TZM-bl cells through infection assay .

Objective To access the incidence,clinical characteristics and the factors affecting therapy of respiratory distress syndrome (RDS) in the infants at term and near term delivered by elective cesarean section.Methods A retrospective cohort study among consecutively admitted infants with RDS at the Neonatal Intensive Care Unit of the Department of Neonatology,Xiangya Hospital,Central South University from Jan.2004 to Dec.2011 were conducted.The inborn infants at 36-42 weeks gestation with RDS,whom were delivered by Elective Cesarean Section from January 1 st,2004 to December 31st,2011 were enrolled.These cases with the timing of elective caesarean section,gestational age,intrauterine infection,asphyxia at birth,which affecting the occurrence of RDS were compared.Results Fifty one infants were entered into the study,which were all met standard of Elective Cesarean Section.Among these infants,33 cases (64.7％,33/51 cases) were delivered by cesarean section without any reason.In these 51 cases,the constituent ratio of elective caesarean section in gestational age ＞ 39 weeks was lower than in gestational age ＞ 36-＜39 weeks,and the difference was significant (31.4％ vs 68.6％,x2 =0.560,P ＜0.01).Asphyxia at birth was the main risk factors of term and near term with RDS (OR =7.306,95％CI:0.018-51.101,P =0.041).Compared to the infants whom born without asphyxia,the infants born with asphyxia usually came out to RDS right after born (x2 =0.080,P ＜ 0.01),required longer time of mechanical ventilation and had significant lower effective ratio (x2 =0.071,8.843,all P ＜ 0.01).Conclusions Asphyxia is the first manifestations of term and near term infants with RDS.These infants often can be onset after birth.

Objective:To establish a fast method for the generation of immature dendritic cells(DCs) from human peripheral blood mononuclear cells(hPBMCs) in vitro.Methods: High purity human CD14+ monocytes were collected using density Ficoll gradient centrifugation and MACS beads sorting system.iDCs(Immature DC) were induced after cultured with rhGM-CSF and rhIL-4 on the fourth day.Fluorescence activated cell sorting(FACS) was used to identify cell surface markers(CCR5) and capabilities of antigen uptake of iDCs on the fourth day.Ordinary optical microscope,scanning electron microscopy and transmission electron microscopy were used to observe surface and internal structure of iDCs on the fourth day of culture conditions.Results: FACS result shows that the purity of CD14+ monocytes collected from hPBMCs were more than 94%.The antigen uptake capability and CD195 of iDCs was detected on the fourth day of cultured conditions.Typical surface and internal structure characteristics of iDCs were observed.Conclusion: Rapid induction culture is an effective method for obtaining a large number of iDC with typical characteristics in vitro,and can be used for further experimental study.

OBJECTIVEToday metal-ceramic restorations are widely accepted in dental practice because of their good aesthetic and biocompatible properties, but there are still several problems to be resolved, among which the influences of their margins on gingival health is noticeable. The purpose of this study was to investigate the influences of metal-ceramic restoration margin on gingival health.

METHODS129 abutments of metal-ceramic restorations, including their margins' fitness, location and smoothness were examined and their influences on gingival health were analyzed.

RESULTSThere was a close relation between the metal-ceramic restorations' margins and the gingival health.

CONCLUSIONIn order to maintain the gingival health, the margins should be smooth, closely fitted, no overhanging and in a moderate location (i.e. not below the gingival more than 1 mm).

Female ; Gingival Diseases ; etiology ; Gingivitis ; etiology ; Hemorrhage ; etiology ; Humans ; Male ; Metal Ceramic Alloys ; adverse effects

Objective To construct a recombinant poxvirus vector vaccine, rVTTδTK-RBD, and to evaluate its safety and immunogenicity. Methods The receptor-binding domain (RBD) gene was synthesized with reference to the gene sequence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and was inserted into the polyclonal site of the self-constructed recombinant plasmid pSTKE, to construct the recombinant poxvirus shuttle vector pSTKE-RBD. This was then transfected into BHK-21 cells pre-infected with the vaccinia virus Tiantan strain (VTT). The recombinant poxvirus rVTTδTK-RBD was successfully obtained after several rounds of fluorescence phage screening. The effect of rVTTδTK-RBD on the body mass of BALB/c mice was detected after immunizing mice by intra-nasal vaccination. The levels of specific and neutralizing antibodies produced by rVTTδTK-RBD on BALB/c mice were analyzed after immunizing mice intramuscularly. The effect of rVTTδTK-RBD on T cell subsets in BALB/c mice was detected by flow cytometry. Results Through homologous recombination, enhanced green fluorescent protein (EGFP) screening marker, and multiple rounds of fluorescent phosphorescence phage screening, a recombinant poxvirus rVTTδTK-RBD, expressing RBD with deletions in the thymidine kinase (TK) gene, was successfully obtained, which was validated by PCR. The in vivo experiments on BALB/c mice showed that rVTTδTK-RBD was highly immunogenic against SARS-CoV-2 and significantly reduced toxicity to the body compared to the parental strain VTT. Conclusion The recombinant poxvirus vaccine rVTTδTK-RBD against SARS-CoV-2 is successfully constructed and obtained, with its safety and immunogenicity confirmed through various experiments.
Animals ; Mice ; SARS-CoV-2/genetics* ; COVID-19 ; Vaccines, Synthetic/genetics* ; Genes, Reporter ; Bacteriophages ; Mice, Inbred BALB C

Objective: To investigate the inhibitory effect of recombinant oncolytic adenovirusAd-Apoptin-hTERTp-E1A(ATV) on luciferase-labeled human lung cancer cells (A549-luc) and human lung cancerA549 cells, and to compare the differences in the inhibitory effect on two cell lines. Methods:ATV was used to infectA549-luc cells andA549 cells respectively. WST-1 and crystal violet staining were used to determine the difference in the inhibitory effect of ATV. Hoechst and Annexin V-FITC/PI staining were used to verify the inhibition mode ofATV. Results: WST-1 and crystal violet staining showed thatATV had significant inhibitory effect on bothA549-luc and A549 cells ( P <0.05). ATV showed significant inhibitory effect on both cells at 24, 48 and 72 h ( P <0.05 or P <0.01), and reached the peak at 72 h; ATV at concentrations of 1, 10 and 100 MOI all showed inhibitory effect on both cells, and reached the peak at 100 MOI. Hoechst staining showed that A549-luc cells and A549 cells infected with ATV showed typical nuclear fragmentation and marginal set. The results of Annexin V-FITC/PI Flow cytometry showed that ATV infection resulted in apoptosis of A549-luc and A549 cells, which was in a time-dependent manner and reached the peak at 72 h( P <0.05 or P <0.01). Conclusion: Insertion of luciferase didn’t significantly change the inhibitory effect and inhibitory mode ofATV onA549-luc cells.ATV exerted its in vitro inhibitory effect onA549-luc and A549 cells by inducing cell apoptosis.

¹⁷⁷Lu- prostate specific membrane antigen (PSMA) radio-ligand therapy has been approved abroad for advanced prostate cancer and has been in several clinical trials in China. Based on domestic clinical practice and experimental data and referred to international experience and viewpoints, the expert group forms a consensus on the clinical application of ¹⁷⁷Lu-PSMA radio-ligand therapy in prostate cancer to guide clinical practice.