Objective To study the effects of the adult T cell leukemia virus type 1(HTLV-1) Tax protein on T lymphocytes self-reactive growth hormone(GH). Methods The expression of the growth hormone protein was measured by Western blot in TaxP and TaxN cells, which expresses HTLV-1 Tax or no. The location of growth hormone in the TaxP and TaxN cells were detected by LSCM(laser scanning confocal microscope). The level of growth hormone mRNA in TaxP and TaxN cells was measured by RT-PCR. The pGL2-GH-luc was transfected in TaxN and TaxP cells by Tfx-50-mediated transfection to assay transcriptional activity. The pNF-κB-luc, pAP-1-luc and pNFAT-luc was transfected into TaxP cells with pcDNA3.0-GH by Tfx50 to test bioactivity of the nuclear transcription factors. Results The mRNA and protein expression of GH could be promoted significantly by Tax. Relative to the TaxN cells, the transcriptional activity of GH was significantly increased about 6.37 times in TaxP cells(P＜0.05). Overexpression of GH can increase the activity of NF-κB about 1.7 times in TaxP cells (P＜0.05). Conclusion GH maybe involve in adult T-cell leukemia(ATL) through activating NF-κB.

To accurately analyze metabolites in industry-important photosynthetic microbes, LC-MS based metabolomics protocol needs to be optimized specifically for individual species. In this study, an LC-MS based metabolomics method was optimized for cyanobacterium Synechocystis sp. PCC 6803. With the optimized extraction, liquid chromatographic and mass spectral parameters, the method was capable of detecting 24 important metabolites related to central carbohydrate and energy metabolism in Synechocystis sp. PCC 6803. The study laid an important foundation for the metabolomics analysis of cyanobacteria.
Chromatography, Liquid ; Mass Spectrometry ; Metabolome ; Metabolomics ; Photosynthesis ; Synechocystis ; metabolism

Objective To explore the expression of early growth response gene-1 (EGR-1) in T cells that were positive for Tax protein of human T-cell leukemia virus type 1 (HTLV-1) and its possible reg-ulatory mechanism .Methods A series of expression structures carrying the regulatory elements of EGR-1 in different length and luciferase reporter genes were constructed .TaxP cells were transfected with the con-structs containing reporter genes and cultured with 5μmol/L of NF-κB inhibitor BAY 11-7082 or equal vol-ume of DMSO.After cultured for 24 hours the cells were collected to test the luciferase activity .BAY 11-7082 or equal volume of DMSO was added into the supernatant of TaxP cell culture to test the expression of EGR-1 protein by Western blot after 24 hours of culture .Tax and its mutants M22 and M47 were transfected into 293 T cells respectively to test the expression of EGR-1 protein by Western blot after 24 hours of culture . Results The expression structures carrying the regulatory elements of EGR-1 in different length and their mutants followed by luciferase reporter genes were successfully constructed .The luciferase activity in the cells transfected with the constructs containing the elements E 1 and E2 were higher than that transfected with E3, DelE and MutE, but the reporter gene expressions were decreased with the interference of BAY 11-7082 (P<0.01).However, there were no significant changes with the luciferase activity in the cells transfected by elements E3, DelE and MutE.Western blot analysis indicated that the expression of EGR-1 protein was significantly decreased with the interference of BAY 10-7082 .The expression of EGR-1 protein in M22 mu-tants-transfected 293 T cells were decreased significantly in comparison with those by wild type tax-and M47-transfected cells .Conclusion NF-κB was the key nuclear factor in regulating the expression of EGR-1 pro-tein in Tax-positive T cells .

Objective To investigate the influence of extracellular high mobility group box 1 (HMGB1) on viral replication in HTLV-1 infected T cells.Methods HMGB1 in culture supernatants of adult T-cell leukemia virus 1 (HTLV-1) virus-negative cell:Jurkat,MOLT4 cells and HTLV-1 virus-positive cells:MT2,MT4,was detected by ELISA;The HTLV-1 long terminal repeat reporter gene (pHTLV-1-LTR-luc) was transfected into MT2 cells by Tfx-50-mediated transfection,and 0.25,0.50,0.75 μg/ml of HMGB1 polyclonal antibody(HMGB1 PcAb) and its isotype control rabbit IgG antibodies,0.03,0.1,0.3 μg/ml rhHMGB1 and its control PBS,were added into culture supernatant respectively,then luciferase activity was detected after 48 h;Similarly,0.25 μg/ml HMGB1 PcAb and the isotype control antibody,0.3 μg/ml rhH-MGB1 and the control PBS were added to the culture supernatant of MT2 cell,the viral gene,pol1,pol2,gag,env,etc,were performed by real-time PCR.Results Culture supernatant HMGB1 levels has no significant difference between HTLV-1 positive cells MT2 and MT4 and the other two virus-negative T cell lines;Compared with isotype control antibody group,the culture supernatant,to which is added 0.25 μg/ml HMGB1 PcAb,can significantly inhibit the HTLV-1-LTR transcriptional activity and suppress the expressions of the viral gene pol1,pol2,gag,env.Compared with the control PBS,0.3 μg/ml rhHMGB1 significantly promotes the transcriptional activity of the HTLV-1-LTR and the expressions of the viral gene pol1,pol2,gag,env.Conclusion The extracellular HMGB1 can promote viral replication of HTLV-1 infected T cells.

Objective     To evaluate the early clinical effect of reimplantation in the treatment of bicuspid aortic valve (BAV) with aortic root aneurysm. Methods     The clinical data of 25 patients with BAV and aortic root aneurysm [mean diameter: 45-63 (52.68±5.55) mm] undergoing reimplantation in West China Hospital from November 2019 to May 2021 were retrospectively reviewed. There were 22 males and 3 females. The mean age was 15-65 (50.00±13.10) years and body surface area was 1.79±0.23 m2. Results    The pathological classification of BAV malformation was confirmed during the operation: Type 0 in 3 patients and Type 1 in 22 patients. There were 12 patients undergoing cusp central plication, and 2 patients were sutured with a closed fusion crest. Postoperative valve leaflet coaptation height was 0.78±0.15 cm, and effective height was 1.27±0.19 cm. In operation, maximum aortic valve flow velocity was 1.65±0.42 m/s, pressure difference was 5.46±3.05 mm Hg, and aortic valve annulus diameter was 21.32±0.95 mm. Cardiopulmonary bypass time was 225.84±35.34 min, and aortic block time was 189.60±26.51 min. In-hospital time was 11.64±3.07 d, ICU stay time was 2.64±0.99 d, and mechanical ventilation time was 1.48±0.87 d. The follow-up time was 17.20±4.70 months, and no death or major complications occurred during the follow-up in all patients. The cardiac function of the patients significantly improved postoperatively (P≤0.05). Echocardiography suggested that 12 patients had no aortic regurgitation, 10 minor aortic regurgitation, 3 mild aortic regurgitation, and no patients with moderate or more severe regurgitation. The diameter of the aortic sinus, left ventricular end-diastolic diameter and volume decreased during the follow-up, compared to preoperative ones (P≤0.05). The maximum flow velocity of the aortic valve was 1.54±0.36 m/s, and the pressure difference was 5.17±2.38 mm Hg during the follow-up. Conclusion    Reimplantation technology has a good clinical effect for highly selective BAV patients. It can effectively avoid long-term postoperative anticoagulation, but the maximum flow rate after surgery is slightly increased, which may be related to the configuration of BAV itself. While compared with valve replacement, the effect is still worthy of recognition.