BACKGROUND:Cationic liposome-mediated celltransfection is reliable and repeatable. However the transfection efficiency is often low.
OBJECTIVE:To study the optimized methods for gene transfection mediated by liposome into N2a cells (mouse neuroblastma cells).
METHODS:Using traditional adherent method and improved suspension method, 500 ng recombinant plasmid pcDNA3-GFP carrying green fluorescence protein was transfected into N2a cells in 24-wel culture plate, which was mediated by 1.5μL Lipofectamine?LTX Reagent. The expression of green fluorescent protein was observed by inverted fluorescence microscope, and the transfection efficiencies at different transfection ways were calculated. By using improved suspension transfection method, 500 ng plasmid DNA was transfected with different doses of Lipofectamine?LTX Reagent (1.0, 1.5, 2.0, 2.5μL). The optimal ratio of liposome and DNA was explored.
RESULTS AND CONCLUSION:The transfection efficiency of suspension transfection method was significantly higher than that of the tranditional adherent method (P<0.01) when using 1.5μL liposome/500 ng DNA. The transfection efficiency of the 1.0, 1.5, 2.0, 2.5μL Lipofectamine?LTX on 500 ng plasmid DNA was respectively (76.60±3.85)%, (80.00±4.17)%, (88.00±5.89)%, (54.96±4.23)%. It showed the 500 ng DNA and 2.0μL liposome achieve the highest transfection efficiency.

Objective: To investigate association between adolescent depressive symptoms with circadian rhythm and emotion regulation strategies, and to provide the basis for mental health education for depression. Methods: CES-D, Morning and Evening Questionnaire-5 (MEQ-5) and Emotion Regulation scale (ERS) were administered to 2 398 students from 6 middle schools in Chengdu, Langzhong and Leshan of Sichuan Province. SPSS 21.0 was used to data processing and anlysis. Results: About 37.9% (909/2 398) of adolescents reported depressive symptoms. Prevalence of depressive symptom was higher in female students( χ 2= 25.15 , P <0.01), rural adolescents( χ 2=15.45, P <0.01), adolescents aged 15-18 compared to aged 12-14( χ 2=187.24, P < 0.01 ). There was significant difference in rate of depressive symptoms among adolescents with different circadian rhythms( χ 2= 55.19 , P < 0.01 ), with definite evening rhythm preference was the highest(57.1%). Prevalence of depressive symptoms significantly varied by sleep duration( χ 2=141.99, P <0.01), and were highest in adolescents with sleep duration <6 h(69.4%). The scores of suppression dimension in depressed adolescents were significantly higher than that of non depressive group, while the scores of reappraisal dimension were significantly higher in non depressive group than that of depressive group. Multivariate Logistic regression analysis showed that gender( OR =1.60), age( OR=2.29), suppression( OR = 1.13 ), sleep duration <6 h( OR =5.17), sleep duration 6-8 h ( OR =2.88) were positively associated with depressive symptoms in adolescents. Moderate type( OR =0.53), morning type ( OR =0.55) and cognitive reappraisal ( OR =0.90) were associated with lower rate of depressive symptoms( P <0.05). Conclusion Sleep rhythm delay, lack of sleep and emotion suppression in adolescents are associated with higher risk for depression. Regular sleep habits and reasonable emotion regulation might help to prevent adolescent depression.

Hydrofluoric acid inhalation injury is difficult to treat, despite it has low incidence. It could cause mild symptoms such as cough and sore throat, or severe symptom that may develop into life-threatening acute respiratory distress syndrome, and even rare pulmonary diseases such as reactive airway dysfunction syndrome and pulmonary alveolar proteinosis. Currently, there is no specific standard for the diagnosis and treatment of hydrofluoric acid inhalation injury. Authors summarize the incidence, injury mechanism, clinical diagnosis and treatment of hydrofluoric acid inhalation injury by searching literature at home and abroad and propose that pulse contour cardiac output monitor and extracorporeal membrane oxygenation have great application prospects in treatment of severe cases, so as to provide references for peers.

Objective:To investigate the efficacy and safety of house dust mite (HDM) allergen subcutaneous specific immunotherapy (SCIT) in patients with allergic rhinitis (AR) with single dust mite allergy and multiple allergen allergy.Methods:A retrospective study was conducted. A total of 372 patients with allergic rhinitis induced by house dust mite were diagnosed in the allergy clinic of General Hospital of North Theater Command from January 2013 to January 2018.They were treated with house dust mite allergen preparation for standardized SCIT for 3 years or more, and had complete follow-up data. The age ranged from 5 to 55 years, the median age was 13 years, and the average age was (19.4±14.7) years; 216 males and 156 females. According to their age, they were divided into the older group (age >14 years) and younger group (age ≤ 14 years). According to the number of allergens, they were divided into single group (only HDM group allergic to house dust mites) and multi recombination (including 2 or more allergens including house dust mites). The multi recombination was further divided into HDM+1 group, HDM+2 group, HDM+3 group, HDM+4 and above group. Before treatment (T0), 1 year (T1) and 3 years (T2) after SCIT treatment, the patients in each group established files, analyzed and compared the average total nasal symptoms score (TNSS), total non nasal symptoms score (TNNSS), visual analogue scale (VAS), total medicine score (TMS) and rhinoconjunctivitis quality of life questionnaire (RQLQ), and evaluated the clinical efficacy of the treatment and the comparison of various scores in the efficacy of SCIT with different allergens and ages. Record the occurrence of local and systemic adverse reactions of all patients during treatment, and evaluate the safety of SCIT. All scores are measurement data that do not conform to normal distribution. Mann-Whitney U and Kruskai-Wallis test of independent samples are used for inter group comparison, and Bonferroni correction is used for further pairwise comparison; Chi square test and continuity correction method were used for the comparison between count data groups such as the incidence of adverse reactions and the effective rate of TNSS, and a-division method was used for further pairwise comparison. Results:After SCIT treatment, the scores of TNSS, TNNSS, TMS, VAS and RQLQ in T1 and T2 were significantly lower than those in T0, and the scores in T2 were significantly lower than those in T1 ( Z=-11.168, -4.786, -6.639, -13.012, -10.652 in T0 vs T1; Z=-13.527, -8.746, -13.397, -14.477, -11.833 in T0 vs T2; Z=-4.721, -4.607, -10.020, -7.180, -5.721 in T1 vs T2; P<0.05). In T1 and T2, compared with the older group, the scores of TNSS, TNNSS, TMS, VAS and RQLQ in younger group were lower, and the differences of various indexes were statistically significant(the median scores of T1: Myounger=3.0, 1.0, 2.0, 4.0, 2.6, Molder=5.0, 2.0, 3.0, 5.0, 3.2; the median scores of T2: Myounger=3.0, 1.0, 0, 2.0, 1.3, Molder=4.0, 1.0, 1.5, 3.0, 2.3; ZT1=-4.525, -5.830, -4.061, -3.608, -2.785; ZT2=-3.847, -4.055, -2.820, -2.998, -3.418; P<0.05). In T1 and T2, the scores of TNSS, VAS and RQLQ in a single group after SCIT treatment were lower than those in multiple recombination(the median scores of T1: Msingle=4.0, 4.0, 2.6, Mmultiple=5.0, 5.0, 3.2; the median scores of T2: Msingle=3.0, 2.0, 1.4, Mmultiple=4.0, 3.0, 2.1), and the difference was statistically significant ( ZT1=-3.002, -2.092, -1.977; ZT2=-3.354, -2.469, -2.116; P<0.05). There was no significant difference in TMS (the median score during T1 period: Msingle=2.0, Mmultiple=3.0, ZT1=-1.130; the median score during T2 period: Msingle=1.0, Mmultiple=1.0, ZT2=-1.544; P>0.05). Further comparison within the group showed that there was no significant difference in the improvement rate of TNSS during T2 period among HDM group, HDM+1 group, HDM+2 group and HDM+3 group (HDM vs HDM+1 group χ 2=0.277, HDM vs HDM+2 group χ 2=0.78, HDM vs HDM+3 group χ 2=0.075, HDM+1 vs HDM+2 group χ 2=0.057, HDM+1 vs HDM+3 group χ 2=0.019, HDM+2 vs HDM+3 group χ 2=0.003; P>0.005), the improvement rates were 92.5%, 90.3%, 89.1% and 89.5%. Respectively in HDM group,HDM+1 group, HDM+2 group, HDM+3 group, compared with HDM+4 and above group, the difference was statistically significant (χ 2=26.144, 13.254, 15.144, 8.808; P<0.005). The improvement rate of TNSS in HDM+4 and above group was 60.9%. 122 patients had local adverse reactions during the treatment of SCIT, accounting for 32.8%. The local adverse reactions were 759 injections (15 336 injections in total), accounting for 4.95%. Most of them were swelling, dizziness, induration and pruritus at the injection site, which could be relieved by oral antihistamines or within 2 hours. There were 2 cases of local urticaria, once for each case. The symptoms were relieved within 1 week after oral antihistamine. No serious systemic adverse reactions occurred. Conclusion:Standardized SCIT may be a safe and effective treatment for AR patients, and the type of allergen may be one of the important factors affecting the efficacy of SCIT. The efficacy of SCIT was significant in AR patients with three or less allergens other than house dust mite.

Objective:To investigate the efficacy and safety of house dust mite (HDM) allergen subcutaneous specific immunotherapy (SCIT) in patients with allergic rhinitis (AR) with single dust mite allergy and multiple allergen allergy.Methods:A retrospective study was conducted. A total of 372 patients with allergic rhinitis induced by house dust mite were diagnosed in the allergy clinic of General Hospital of North Theater Command from January 2013 to January 2018.They were treated with house dust mite allergen preparation for standardized SCIT for 3 years or more, and had complete follow-up data. The age ranged from 5 to 55 years, the median age was 13 years, and the average age was (19.4±14.7) years; 216 males and 156 females. According to their age, they were divided into the older group (age >14 years) and younger group (age ≤ 14 years). According to the number of allergens, they were divided into single group (only HDM group allergic to house dust mites) and multi recombination (including 2 or more allergens including house dust mites). The multi recombination was further divided into HDM+1 group, HDM+2 group, HDM+3 group, HDM+4 and above group. Before treatment (T0), 1 year (T1) and 3 years (T2) after SCIT treatment, the patients in each group established files, analyzed and compared the average total nasal symptoms score (TNSS), total non nasal symptoms score (TNNSS), visual analogue scale (VAS), total medicine score (TMS) and rhinoconjunctivitis quality of life questionnaire (RQLQ), and evaluated the clinical efficacy of the treatment and the comparison of various scores in the efficacy of SCIT with different allergens and ages. Record the occurrence of local and systemic adverse reactions of all patients during treatment, and evaluate the safety of SCIT. All scores are measurement data that do not conform to normal distribution. Mann-Whitney U and Kruskai-Wallis test of independent samples are used for inter group comparison, and Bonferroni correction is used for further pairwise comparison; Chi square test and continuity correction method were used for the comparison between count data groups such as the incidence of adverse reactions and the effective rate of TNSS, and a-division method was used for further pairwise comparison. Results:After SCIT treatment, the scores of TNSS, TNNSS, TMS, VAS and RQLQ in T1 and T2 were significantly lower than those in T0, and the scores in T2 were significantly lower than those in T1 ( Z=-11.168, -4.786, -6.639, -13.012, -10.652 in T0 vs T1; Z=-13.527, -8.746, -13.397, -14.477, -11.833 in T0 vs T2; Z=-4.721, -4.607, -10.020, -7.180, -5.721 in T1 vs T2; P<0.05). In T1 and T2, compared with the older group, the scores of TNSS, TNNSS, TMS, VAS and RQLQ in younger group were lower, and the differences of various indexes were statistically significant(the median scores of T1: Myounger=3.0, 1.0, 2.0, 4.0, 2.6, Molder=5.0, 2.0, 3.0, 5.0, 3.2; the median scores of T2: Myounger=3.0, 1.0, 0, 2.0, 1.3, Molder=4.0, 1.0, 1.5, 3.0, 2.3; ZT1=-4.525, -5.830, -4.061, -3.608, -2.785; ZT2=-3.847, -4.055, -2.820, -2.998, -3.418; P<0.05). In T1 and T2, the scores of TNSS, VAS and RQLQ in a single group after SCIT treatment were lower than those in multiple recombination(the median scores of T1: Msingle=4.0, 4.0, 2.6, Mmultiple=5.0, 5.0, 3.2; the median scores of T2: Msingle=3.0, 2.0, 1.4, Mmultiple=4.0, 3.0, 2.1), and the difference was statistically significant ( ZT1=-3.002, -2.092, -1.977; ZT2=-3.354, -2.469, -2.116; P<0.05). There was no significant difference in TMS (the median score during T1 period: Msingle=2.0, Mmultiple=3.0, ZT1=-1.130; the median score during T2 period: Msingle=1.0, Mmultiple=1.0, ZT2=-1.544; P>0.05). Further comparison within the group showed that there was no significant difference in the improvement rate of TNSS during T2 period among HDM group, HDM+1 group, HDM+2 group and HDM+3 group (HDM vs HDM+1 group χ 2=0.277, HDM vs HDM+2 group χ 2=0.78, HDM vs HDM+3 group χ 2=0.075, HDM+1 vs HDM+2 group χ 2=0.057, HDM+1 vs HDM+3 group χ 2=0.019, HDM+2 vs HDM+3 group χ 2=0.003; P>0.005), the improvement rates were 92.5%, 90.3%, 89.1% and 89.5%. Respectively in HDM group,HDM+1 group, HDM+2 group, HDM+3 group, compared with HDM+4 and above group, the difference was statistically significant (χ 2=26.144, 13.254, 15.144, 8.808; P<0.005). The improvement rate of TNSS in HDM+4 and above group was 60.9%. 122 patients had local adverse reactions during the treatment of SCIT, accounting for 32.8%. The local adverse reactions were 759 injections (15 336 injections in total), accounting for 4.95%. Most of them were swelling, dizziness, induration and pruritus at the injection site, which could be relieved by oral antihistamines or within 2 hours. There were 2 cases of local urticaria, once for each case. The symptoms were relieved within 1 week after oral antihistamine. No serious systemic adverse reactions occurred. Conclusion:Standardized SCIT may be a safe and effective treatment for AR patients, and the type of allergen may be one of the important factors affecting the efficacy of SCIT. The efficacy of SCIT was significant in AR patients with three or less allergens other than house dust mite.

The operating room was the core department of a hospital,and its operational efficiency had a significant impact on the high-quality development of a hospital.An analysis has revealed that low efficiency and irrational allocation in the operating room were mainly due to the lack of operational regulations and norms,the unreasonable arrangement of surgical specialties,and the unbalanced allocation of supporting resources.To address these issues,the First Affiliated Hospital of Zhengzhou University has taken into account the overall allocation of resources for the central operating room and the central operating room,and formulated strategies to improve operational efficiency,including adjusting the operational mechanism,optimizing the structure of surgical specialties,and providing corresponding supporting resources.Based on the adjustment of surgical structure,the implementation effect of the program was measured and evaluated,which provided practical strategies for optimizing operating room efficiency in hospitals.

The operating room was the core department of a hospital,and its operational efficiency had a significant impact on the high-quality development of a hospital.An analysis has revealed that low efficiency and irrational allocation in the operating room were mainly due to the lack of operational regulations and norms,the unreasonable arrangement of surgical specialties,and the unbalanced allocation of supporting resources.To address these issues,the First Affiliated Hospital of Zhengzhou University has taken into account the overall allocation of resources for the central operating room and the central operating room,and formulated strategies to improve operational efficiency,including adjusting the operational mechanism,optimizing the structure of surgical specialties,and providing corresponding supporting resources.Based on the adjustment of surgical structure,the implementation effect of the program was measured and evaluated,which provided practical strategies for optimizing operating room efficiency in hospitals.

The operating room was the core department of a hospital,and its operational efficiency had a significant impact on the high-quality development of a hospital.An analysis has revealed that low efficiency and irrational allocation in the operating room were mainly due to the lack of operational regulations and norms,the unreasonable arrangement of surgical specialties,and the unbalanced allocation of supporting resources.To address these issues,the First Affiliated Hospital of Zhengzhou University has taken into account the overall allocation of resources for the central operating room and the central operating room,and formulated strategies to improve operational efficiency,including adjusting the operational mechanism,optimizing the structure of surgical specialties,and providing corresponding supporting resources.Based on the adjustment of surgical structure,the implementation effect of the program was measured and evaluated,which provided practical strategies for optimizing operating room efficiency in hospitals.

The operating room was the core department of a hospital,and its operational efficiency had a significant impact on the high-quality development of a hospital.An analysis has revealed that low efficiency and irrational allocation in the operating room were mainly due to the lack of operational regulations and norms,the unreasonable arrangement of surgical specialties,and the unbalanced allocation of supporting resources.To address these issues,the First Affiliated Hospital of Zhengzhou University has taken into account the overall allocation of resources for the central operating room and the central operating room,and formulated strategies to improve operational efficiency,including adjusting the operational mechanism,optimizing the structure of surgical specialties,and providing corresponding supporting resources.Based on the adjustment of surgical structure,the implementation effect of the program was measured and evaluated,which provided practical strategies for optimizing operating room efficiency in hospitals.

The operating room was the core department of a hospital,and its operational efficiency had a significant impact on the high-quality development of a hospital.An analysis has revealed that low efficiency and irrational allocation in the operating room were mainly due to the lack of operational regulations and norms,the unreasonable arrangement of surgical specialties,and the unbalanced allocation of supporting resources.To address these issues,the First Affiliated Hospital of Zhengzhou University has taken into account the overall allocation of resources for the central operating room and the central operating room,and formulated strategies to improve operational efficiency,including adjusting the operational mechanism,optimizing the structure of surgical specialties,and providing corresponding supporting resources.Based on the adjustment of surgical structure,the implementation effect of the program was measured and evaluated,which provided practical strategies for optimizing operating room efficiency in hospitals.