1.Salivary protein rAlb-34kDa-1 of Aedes albopictus modulates the immune response of Raw264.7 cells and promotes the replication of DENV-2
KUANG Xiaoyuan ; XIAO Qiuqiu ; WU Wenshi
China Tropical Medicine 2025;25(2):149-
Objective To investigate the immunomodulatory effects of Aedes albopictus salivary protein rAlb-34kDa-1 on macrophages (Raw264.7) and its impact on dengue virus type 2 (DENV-2) replication in Raw264.7 cells. Methods The prokaryotic expression plasmid pET32a(+)-rAlb-34kDa-1 was transfected into Escherichia coli (E.coli) BL21 (DE3), and salivary proteins were obtained according to the expression purification conditions established in the pre-laboratory stage and identified by SDS-PAGE and Western blot (WB). The toxic effects of rAlb-34kDa-1 protein in Raw264.7 cells were assessed by CCK8 assay. Raw264.7 cells were treated with rAlb-34kDa-1 at concentrations of 0.02, 0.2, and 2 μg/mL for 6, 12, and 24 h. The expression of interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), chemokine ligand 2 (CCL2), interferon-β (IFN-β), and interferon-stimulated gene 15 (ISG15) within the cells was detected using real-time quantitative reverse transcription PCR (qRT-PCR). Enzyme-linked immunosorbent assay (ELISA) was used to measure IL-6 and TNF-α secretion in the cell culture supernatants. Immunofluorescence staining was used to observe the translocation of p65 into the nucleus of Raw264.7 cells following rAlb-34kDa-1 treatment, to analyze whether rAlb-34kDa-1 modulates the immune response in Raw264.7 cells through NF-κB signaling pathway. Cells were collected 6, 12, and 24 hours post-treatment under different experimental conditions (DENV-2 treatment group, salivary protein +DENV-2 co-incubation group) was measured using qRT-PCR, and the fluorescence intensity of J2 (dsRNA) in the cells was detected by flow cytometry (FCM) after 24 hours under different treatment factors. Results The results of SDS-PAGE and WB confirmed the successful acquisition of rAlb-34kDa-1 protein, and CCK8 analysis showed no cytotoxicity of this protein toward Raw264.7 cells. Both 0.2 μg/mL and 2 μg/mL rAlb-34kDa-1 induced the expression of IL-1β, IL-6, TNF-α, CCL2, IFN-β, and ISG15 in Raw264.7 cells. One hour after rAlb-34kDa-1 treatment, the fluorescence intensity of p65 in the nucleus of Raw264.7 cells was significantly enhanced. In Raw264.7 cells co-incubated with rAlb-34kDa-1 and DENV-2 for 12 hours, rAlb-34kDA-1 at a concentration of 2 μg/mL significantly increased the mRNA expression of DENV-2. After 24 hours of treatment, rAlb-34kDA-1 at concentrations of 0.02 μg/mL, 0.2 μg/mL, and 2 μg/mL all enhanced the mRNA expression of DENV-2 in Raw264.7 cells, and 2 μg/mL of rAlb-34kDa-1 co-incubated with DENV-2 significantly increased the J2 fluorescence signal in cells. Conclusion The rAlb-34kDa-1 protein can regulate the immune response in murine Raw264.7 cells and promote the replication of DENV-2 in these cells.
2.Correlation analysis of psychological distress, perceived stress and nurses' caring ability in patients after pulmonary nodule surgery
Hong KUANG ; Huiru HOU ; Xiaoyuan HUYAN
Chinese Journal of Modern Nursing 2022;28(30):4192-4196
Objective:To investigate the current situation of psychological distress in patients after pulmonary nodule surgery and to explore the effect path of perceived stress and nurses' caring ability on psychological distress.Method:From October 2019 to May 2021, a total of 415 patients with pulmonary nodules after surgery in the First Medical Center of Chinese PLA General Hospital were selected as the investigation objects by the convenient sampling method. The Psychological Distress Thermometer, Caring Ability Scale, Perceived Stress Scale and General Information Questionnaire were used for investigation and analysis. Structural equation model was used to analyze the effect of perceived stress on psychological distress. A total of 415 paper questionnaires were distributed and 411 valid questionnaires were recovered, with an effective recovery rate of 99.04%.Results:The psychological distress score of 411 patients with pulmonary nodules after operation was (4.82±1.42) , and 179 patients (43.55%) had a score greater than or equal to 4. There were statistically significant differences in the psychological distress scores of inpatients with different marriage, pulmonary nodule types, fear of cancer, emotional self-regulation ability, e-health literacy and disease awareness after pulmonary nodule surgery ( P<0.01) . Psychological distress was negatively correlated with nurses' caring ability ( r=-0.626, P<0.01) , and positively correlated with perceived stress ( r=0.596, P<0.01) . The mediating effect value of nurse' caring ability between sense of loss of control and psychological pain was 0.132, accounting for 32.84% (0.132/0.402) of the total effect. Conclusions:The psychological pain of patients after pulmonary nodule surgery is closely related to perceived pressure and nurses' caring ability. The nurses' caring ability has a partial mediating effect between the sense of loss of control and psychological pain. Improving nurses' caring ability can reduce the psychological distress of patients after pulmonary nodule surgery.
3.Application of preoperative eye position training in FL-LASIK
Qihong HAO ; Min JIN ; Lihui KUANG ; Xiaoyuan WANG ; Yuehong ZHANG
Modern Clinical Nursing 2013;(5):19-22
Objective To investigate the clinical effects of preoperative eye position training on femtosecond laser-assisted laser in situ keratomileusis(FL-LASIK)? Methods One hundred and sixty-eight myopia patients(328 eyes)scheduled for selective FL-LASIK were randomly divided into the experiment group of 85 cases(166 eyes)and the control group of 83 cases(162 eyes)?The patients in the control group were given the routine preoperative education and the patients in the experiment group received the preoperative intervention of eye position besides routine preoperative education? The two groups were compared in terms of intraoperative changes of eye position,corneal topography and the postoperative visual acuity? Results The frequency of eye position adjustment and machine stopping in the experiment group was significantly smaller than that of the control group(P < 0?001)? But no significant differences were found between the two groups in corneal topography and the postoperative visual acuity(P > 0?05)? Conclusion Preoperative training of eye position may maintain ideal eye position and reduce the frequency of downtime due to eye position adjustment during FL-LASIK,which may ensure the successful completion of FL-LASIK?

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