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.

Objective To investigate the demand of health management of psychiatric patients primary caregivers after discharge, and to provide references for the health management of patients after discharge. Methods A self-designed questionnaire was used.One hundred and eighty two primary caregivers of patients to be discharged in psychiatric department were enrolled.Results 96.15% of caregivers had demands of health management for patients after discharge.In the content of management, the highest score was relapse prevention related knowledge (4.75 ±0.23), the lowest score was the vocational skills training (2.74 ±0.47).The highest demand of health management mode of patients after discharge from hospital was telephone follow-up, totally 158 caregivers (86.81%), followed by medical personnel on-site guidance, totally 139 caregivers (76.37%).In the management of frequency, most of the caregivers selected telephone and SMS follow-up once per half a month, or medical personnel on-site guidance once per two months;or designated medical expert consultants once per month. In the management period, 3 to 6 months after discharge was the mainly choice.Conclusions The main caregivers of patients after discharge from hospital has strong demand of health management.The hospital should establish reasonable health management service system according to the needs and wishes of patients, and carry out targeted content rich, diverse forms, a specific number of health management services, to improve the patient’ s family function, promote the recovery of social function of patients, to reduce the relapse rate and disabled rate.