OBJECTIVETo construct eukaryotic expression plasmid IRES-gD-IL-2 which contains both HSV-1 glycoprotein D (gD) gene and IL-2 gene and to induce it to express in antigen presenting cell (APC) -- dendritic cells (DCs).

METHODSThe whole sequence of gD and IL-2 were amplified by PCR assay. After confirmation by PCR, double-enzyme digestion and sequencing, these genes were directly cloned into eukaryotic expression vector IRES, then were transfected into DCs. Western blotting was employed to identify the transcription and expression of gD gene.

RESULTSThe results of PCR and enzyme digestion showed that the recombinant expression plasmid contained correct fragments, and the transcription and expression of gD were confirmed by Western blotting.

CONCLUSIONThe recombinant expression vector IRES-gD-IL-2 was constructed, the results of the Western blotting showed that the recombinant protein could be identified by gD- specific antibody, therefore the protein has immunologic competence.

Blotting, Western ; Dendritic Cells ; metabolism ; Herpes Simplex Virus Vaccines ; immunology ; Humans ; Interleukin-2 ; genetics ; Plasmids ; Recombinant Fusion Proteins ; genetics ; immunology ; Vaccines, DNA ; immunology ; Viral Envelope Proteins ; genetics

Adult ; CD4 Lymphocyte Count ; Cell Line, Tumor ; Female ; HIV-1 ; classification ; physiology ; Humans ; Male ; Middle Aged ; Receptors, CCR5 ; physiology ; Receptors, CXCR4 ; physiology ; Recombination, Genetic ; Viral Load

Aneurysms and pseudoaneurysm are commonly encountered with arteriovenous vascular access for haemodialysis. They are difficult complications to manage. Due to the limited number of vascular centers available, patients were unable to seek treatment until further complications arise. The technique of partial aneurysmectomy and primary repair was adopted as the method of repair in this study. A total of 20 cases underwent the surgery from 2019 to 2020. Among the 20 cases, one patient had two pseudoaneurysms at different location which requires her to undergo the procedure twice. The successful cannulation rate post repair was 70% whereas the overall complication rate was 35% which might be attributed to the small sample size. Overall, this study demonstrated that AVF aneurysm and pseudoaneurysm can be safely treated with this approach and can be done effectively in non-vascular centers by general surgeons.

A 7-year-old well and healthy boy was hospitalized for the first time for severe respiratory distress. He was diagnosed with congenital cystic adenomatoid malformation (CCAM) and bronchopulmonary sequestration (BPS). CCAM and BPS are rare congenital pulmonary diseases and their presentation concurrently as a hybrid lesion is even rarer. CCAM is usually diagnosed prenatally however it may regress postnatally and the patient may be asymptomatic. BPS usually presents after birth or later in childhood with recurrent lung infections. We highlight this case of hybrid lesion of CCAM and BPS with an atypical clinical presentation which was lifesaving surgically with diagnosis made via radiographic imaging.
Bronchopulmonary Sequestration

OBJECTIVE： To study the effects of benzoyl aconitine （BAC） on autophagy and apoptosis of human lung cancer A549 cells， and to investigate its mechanism in anti-non-small cell lung cancer. METHODS： A549 cells were treated with different doses of BAC （10， 50， 100， 200， 400 μmol/L）， and then cell morphology was obtained； the proliferation inhibition rate of the cell was determined by CCK-8 assay. The cells were divided into control group （without drug）， BAC low-dose and high-dose groups （200， 400 μmol/L）. After treated with relevant drugs， the apoptosis rate of cells was determined by flow cytometry. The gene and protein expression of apoptosis-related factors Bcl-2， Bax， Caspase-3 as well as autophagy-related factors Beclin1， LC3， P62 were determined by RT-PCR and Western blotting assay. RESULTS： After treated with different doses of BAC， the cells were shrunken and sparsely arranged； inhibitory rate of cell proliferation was increased significantly in BAC 100， 200， 400 μmol/L groups （P＜0.05 or P＜0.01）. Results of flow cytometry showed that the apoptotic rates of cells were increased to different extents in BAC low-dose and high-dose groups after treated for 24 and 48 h， in a concentration and time-dependent manner. Compared with control group， mRNA and protein expression of Bcl-2 and P62 were decreased to different extents in BAC groups； mRNA expression of Bax， Caspase-3， Beclin1 and LC3 as well as protein expression of Bax， Active caspase-3， P62， Beclin1， LC3Ⅱ/Ⅰ were increased to different extent； there was statistical significance in mRNA expression of Caspase-3， and protein expression of Bcl-2， Active Caspase-3， Beclin1， LC3Ⅱ/Ⅰ and P62 in BAC low-dose group as well as all target mRNA and protein expression in BAC high-dose group （P＜0.05 or P＜0.01）， in dose-dependent manner. CONCLUSIONS： BAC can inhibit the proliferation and promote the apoptosis of A549 cells， promote Beclin1， LC3（LC3Ⅱ/Ⅰ），Bax and Caspase-3 （Active Caspase-3） gene and their protein expression， but inhibit P62 and Bcl-2 gene and their protein expression. The mechanism may be related to BAC inducing apoptosis by promoting excessive autophagy of cells.