OBJECTIVETo evaluate the anti-viral effects of a plasmid expressing an inverted-repeat RNA targeting dengue virus type-2 (DENV-2) pre-membrane (prM) gene.

METHODSuckling mice were inoculated with live DENV-2 in the brain. The total RNA was extracted from the brain tissue of the infected mice, and the prM gene fragments were amplified by RT-PCR and then subcloned into XhoI/EcoR I of the pcDNA3.1(+) plasmid in antisense orientation to construct the plasmid pcDNA-asprM. DENV-2 prM sequences were also subcloned into pMD18-T-vector in sense orientation to construct the plasmid pMD18-T- prM. pcDNA-irRNA was constructed by inserting in sense orientation the prM fragment isolated from pMD18-T-prM into the NheI/Kpn I of pcDNA-asprM. The plasmid pcDNA-irRNA was transfected into BHK-21 cells and the anti-viral effects were analyzed by semi-quantitative PCR and real-time PCR.

RESULTSTransfection with the plasmid pcDNA-irRNA caused a reduction of NS3 mRNA expression level by 28% in BHK-21 cells following a 96-h challenge with DENV-2 as compared to the cells without plasmid transfection (positive control). The viral copies in pcDNA-irRNA-transfected cells was 1.44-fold lower than those in the positive control cells following a 72-h virus challenge, and the mRNA expression levels of NS1 were also significantly lower in the transfected cells at 96 h after viral challenge (P<0.05) as shown by real-time quantitative PCR.

CONCLUSIONThe inverted-repeat RNA for DENV-2 prM gene silencing can suppress DENV-2 replication in BHK-21 cells, which provides a basis for developing dengue virus gene vaccine.

Animals ; Base Sequence ; Cells, Cultured ; Cricetinae ; Dengue Virus ; physiology ; Gene Silencing ; Mice ; Mice, Inbred Strains ; RNA, Viral ; genetics ; Terminal Repeat Sequences ; Viral Envelope Proteins ; genetics ; Virus Replication ; genetics

Objective To investigate the effect of perceived service quality improvement during reasonable use of intraoperative period among patient′s relatives by the application of process management model in Ophthalmology Department.Methods We investigated 100 relatives of patients who had surgery from July to September in 2013 by questionnaire.The investigation included the appropriate time to educate health knowledge and satisfaction degrees.Results The relatives′satisfaction degree was beyond 95% at one-to-one tutorials, repeated surgical knowledge on TV, the process of changing and attitude of medical staff in waiting time.For the appropriate time to educate, 70%of relatives believed the waiting time was the right time, but 27%of them preferred acquired after operation, and 3%of them had indifference of time.Conclusions Process management model plays a significant role to improve the perceived service quality of the relatives during the waiting time when the patients are having surgeries in Ophthalmology Department.

Objective To construct a postoperative lung infection preventive care program for lung cancer patients and carry out preliminary validation,so as to provide a reference basis that can be used in clinical practice.Methods Literature analysis was carried out to search domestic and international databases,guideline networks,and association websites,with a search timeframe of January 2011 to December 2021,to obtain relevant evidence,to construct a preliminary draft of a preventive care program for postoperative lung infections in patients with lung cancer,and to use the Delphi method of correspondence with experts from Guangdong Province,Henan Province,and Sichuan Province,September-December 2022,to determine the content of the program.Correspondence to determine the content of the program.In November-December 2023,15 thoracic surgery nurses and 15 lung cancer patients in a tertiary hospital in Shantou City were selected for the initial application of the program and evaluated the effectiveness of the program through the nurses'clinical significance,applicability,feasibility,and patient satisfaction.Results A total of 21 experts completed 2 rounds of correspondence.95.65％and 95.45％of questionnaires were retumed in the 2 rounds of correspondence;the authority coefficients of the experts were 0.941 and 0.948;the Kendall's harmony coefficients were 0.176 and 0.310;the coefficients of variation for each item in the 2nd round of correspondence were 0.04～0.25.The final draft of the program included 10 aspects and 37 measures,such as teamwork,health education,risk assessment,rehabilitation training,fluid management,oral manage-ment,airway management,nutritional management,pain management,line management,and other 10 areas with 37 measures.After the initial application of the program,100％of the nurses thought that the program was meaningful,87％of the nurses thought that the program was clinically applicable and feasible,and 100％of the patients indicated that they were satisfied with the program,of which 80％were very satisfied.Conclusion The constructed nursing program for preventing postoperative lung infection in lung cancer patients is scientific,relevant and feasible,and can provide references for clinical health care personnel to develop measures for preventing postoperative lung infection in lung cancer patients.

Voice recognition is a key skill for successful human communication,but patients with voice recog-nition defects often suffer from the inability to accurately identify the speaker's identity.Phonagnosia refers to the difficulty in identifying a speaker by voice alone when hearing ability is not impaired.From the perspective of brain injury,the symptoms can be divided into congenital phonagnosia and developmental phonagnosia,and congenital phonagnosia is related to the dysfunction of the functional connection between the temporal lobe and the amygdala.The damaged brain areas in patients with acquired speech recognition defect mainly include the temporal lobe and the frontal lobe.In the future,more attention should be paid to the screening methods,neural mechanisms and differ-ences between patients with speech recognition deficiency and other patients with hearing and cognitive impairment.