OBJECTIVE: To establish a rapid detection and genotyping method for SARS-CoV-2 Omicron BA.4/5 variants using CRISPPR-Cas12a gene editing technology. METHODS: We combined reverse transcription-polymerase chain reaction (RT-PCR) and CRISPR gene editing technology and designed a specific CRISPPR RNA (crRNA) with suboptimal protospacer adjacent motifs (PAM) for rapid detection and genotyping of SARS- CoV-2 Omicron BA.4/5 variants. The performance of this RT- PCR/ CRISPPR-Cas12a assay was evaluated using 43 clinical samples of patients infected by wild-type SARS-CoV-2 and the Alpha, Beta, Delta, Omicron BA. 1 and BA. 4/5 variants and 20 SARS- CoV- 2-negative clinical samples infected with 11 respiratory pathogens. With Sanger sequencing method as the gold standard, the specificity, sensitivity, concordance (Kappa) and area under the ROC curve (AUC) of RT-PCR/CRISPPR-Cas12a assay were calculated. RESULTS: This assay was capable of rapid and specific detection of SARS- CoV-2 Omicron BA.4/5 variant within 30 min with the lowest detection limit of 10 copies/μL, and no cross-reaction was observed in SARS-CoV-2-negative clinical samples infected with 11 common respiratory pathogens. The two Omicron BA.4/5 specific crRNAs (crRNA-1 and crRNA-2) allowed the assay to accurately distinguish Omicron BA.4/5 from BA.1 sublineage and other major SARS-CoV-2 variants of concern. For detection of SARS-CoV-2 Omicron BA.4/5 variants, the sensitivity of the established assay using crRNA-1 and crRNA-2 was 97.83% and 100% with specificity of 100% and AUC of 0.998 and 1.000, respectively, and their concordance rate with Sanger sequencing method was 92.83% and 96.41%, respectively. CONCLUSION By combining RT-PCR and CRISPPR-Cas12a gene editing technology, we successfully developed a new method for rapid detection and identification of SARS-CoV-2 Omicron BA.4/5 variants with a high sensitivity, specificity and reproducibility, which allows rapid detection and genotyping of SARS- CoV-2 variants and monitoring of the emerging variants and their dissemination.
Humans ; COVID-19 ; CRISPR-Cas Systems ; Genotype ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction ; SARS-CoV-2/genetics* ; RNA ; COVID-19 Testing

Hepatosteatosis is characterized by abnormal accumulation of triglycerides(TG),leading to prolonged and chronic inflammatory infiltration.To date,there is still a lack of effective and economical therapies for hepatosteatosis.Oridonin(ORI)is a major bioactive component extracted from the traditional Chinese medicinal herb Rabdosia rubescens.In this paper,we showed that ORI exerted significant protective ef-fects against hepatic steatosis,inflammation and fibrosis,which was dependent on LXRα signaling.It is reported that LXRα regulated lipid homeostasis between triglyceride(TG)and phosphatidylethanol-amine(PE)by promoting ATCL and EPT1 expression.Therefore,we implemented the lipidomic strategy and luciferase reporter assay to verify that ORI contributed to the homeostasis of lipids via the regulation of the ATGL gene associated with TG hydrolysis and the EPT1 gene related to PE synthesis in a LXRα-dependent manner,and the results showed the TG reduction and PE elevation.In detail,hepatic TG overload and lipotoxicity were reversed after ORI treatment by modulating the ATCL and EPT1 genes,respectively.Taken together,the data provide mechanistic insights to explain the bioactivity of ORI in attenuating TG accumulation and cytotoxicity and introduce exciting opportunities for developing novel natural activators of the LXRα-ATGL/EPT1 axis for pharmacologically treating hepatosteatosis and metabolic disorders.

Objective:To evaluate the efficacy of combination of toy and cartoon video-assisted admission in preventing agitation during recovery from general anesthesia in preschool children.Methods:Totally 96 children who undergoing elective surgery from July 2018 to October 2019 in Guangzhou Women and Children′s Medical Center were divided into experimental group ( n=48) and control group ( n=48). Two groups were admitted to the general preoperative waiting room for 60 minutes before the operation, and children in the experimental group recieved toy and cartoon video combination intervention, while children in the control group were subjected to routine nursing intervention. The children′s agitation was assessed by using Pediatric Anesthesia Emergence Delirium Scale (PAED) when children entered the post-anesthesia care unit (PACU), 10 minutes after in PACU and leave PACU. In addition, the vital signs such as blood pressure, heart rate, respiratory rate of the two groups were also recorded. Results:The emergence time and duration of PACU stay in the experimental group were (17.19±1.76) minutes and (22.47±2.36) minutes, significantly shortern than in the control group (18.36±2.19) minutes and (23.82±2.62) minutes, the difference was statistically significant ( t values were 2.760, 2.546, P<0.05). 10 minutes after in PACU and out of PACU, the PAED scores, systolic pressure, heart rate, respiratory rate was [7.56±1.86, 4.60±1.19, (108.12±10.19)mmHg, (102.72±8.55) mmHg, (112.91±6.10) times/minute, (106.39±6.75) times/minute, (18.87±1.98) times/minute, (17.49±1.68) times/minute], significantly lower than those in the control group [13.40±1.47, 6.71±2.21, (116.73±14.17)mmHg, (107.62±12.44) mmHg, (121.07±8.07) times/minute, (114.69±5.78) times/minute, (20.96±1.86) times/minute, (18.47±1.63) times/minute], the difference was statistically significant ( t values were 2.162-16.332, P<0.05). Conclusion:Toy and cartoon video combine-assisted admission can reduce the agitation and stable vital signs during recovery from general anesthesia in preschool children.

Objective:To investigate the quality of life of senior women with urinary incontinence and to explore its influencing factors.Methods:A convenient sampling method was used to select 152 senior women with urinary incontinence in Fuzhou Hot-Spring Street Community, Dongjie Street Community and Old-age Care Institutions from June 2017 to September 2017. The quality of life of elderly women with urinary incontinence were observed and the influencing factors were analyzed via International Consultation on Incontinence Questionnaire Short Form (ICI-Q-SF) and Incontinence Quality of Life Scale (I-QOL).Results:The quality of life of senior women with urinary incontinence was (98.7±12.2) points. The scores of the three dimensions including restrictive behavior, social activity limitation and psychological impact were 34.5±4.4 and 41.4±5.4 and 20.8 ± 3.5, respectively. There were 4 factors that influencing the quality of life of patients, namely, the severity of urinary incontinence, the type of urinary incontinence, diabetes and the frequency of leaking urine ( P<0.01). Conclusions:Urinary incontinence has a certain negative impact on the quality of life of patients, and its influencing factors are complex. Individualized care measures should be taken based on the influencing factors to improve the quality of life of patients.

OBJECTIVE:To evaluate the effectiveness and safety of repaglinide combined with metformin versus glimepiride combined with metformin in the treatment of type 2 diabetes mellitus (T2DM),and to provide evidence-based reference for the clinic. METHODS:Retrieved from CJFD, VIP, Wanfang database, PubMed, Embase, Medline and Cochrane Library, randomized controlled trials (RCTs) about therapeutic efficacy (HbA1c, FPG, 2 hPG) and safety (the incidence of ADR,hypoglycemia and gastrointestinal reaction)of repaglinide combined with metformin(trial group)versus glimepiride combined with metformin(control group)in the treatment of T2DM were collected. Meta-analysis was performed by using Rev Man 5.2 statistical software after data extraction and quality evaluation with Cochrane systematic evaluation manual. RESULTS:A total of 12 RCTs were included,involving 957 patients. Results of Meta-analysis showed that the decrease of 2 hPG in trial group was significantly better than control group,with statistical significance [MD=-0.70,95％CI(-1.02,-0.38),P<0.001]. There was no statistical significance in the decrease of HbA1c [MD=0.00,95％CI(-0.24,0.25),P=0.98] or FPG [MD=0.10,95％CI(-0.17,0.36),P=0.47],the incidence of ADR [OR=0.54,95％CI(0.28,1.06),P=0.07],hypoglycemia [OR=0.52,95％CI(0.13,2.06),P=0.35] or gastrointestinal reactions [OR=0.60,95％CI(0.15,2.41),P=0.47] between 2 groups. CONCLUSIONS:Repaglinide combined with metformin is better than glimepiride combined with metformin in reducing 2 hPG,but both of them have similar safety.

OBJECTIVETo investigate the effects of siRNAs targeting CD97 immune epitopes on proliferation, infiltration, apoptosis and cell cycle of breast cancer cells.

METHODSsiRNA sequences targeting CD97and CD97immune epitopes were designed according to Gene Bank NM_001025160.2 with smart siCatchsiRNA design software. CD97siRNAs were transfected into MDA-MB231 cells in which CD97 was highly expressed. Highest sensitive CD97and CD97siRNA were screened by Western blotting. Inverted microscope was used to observe the growth of CD97siRNAs-transfected MDA-MB231 cells; the proliferation activity of MDA-MB231 cells was detected by MTT method; the wound healing assay and Transwell migration test were performed to examine the migration and infiltration ability of CD97and CD97siRNA-transfected MDA-MB231 cells; the effects of CD97siRNA and CD97siRNA on cell apoptosis and cell cycle of MDA-MB231 cells were detected by TUNEL and flow cytometry.

RESULTSThe growth and proliferation activity of CD97siRNAs-transfected MDA-MB231 cells were significantly lower than those in the control groups, and such differences were more significant in CD97siRNA-transfected group (all<0.05); scratch test showed that the wound healing rate was lower in CD97siRNAs-transfected groups, especially in CD97siRNA-transfected group (all<0.05); Transwell migration showed that the number of MDA-MB231 cells crossing through chambers were less in CD97siRNAs-transfected groups, especially in CD97siRNA-transfected group (all<0.05); no significant difference in cell apoptosis was observed between CD97siRNAs-transfected groups and control groups; cell cycle detection showed that CD97siRNAs-transfected groups had less cells in G/Gphase and more cells in S phase compared with the control groups, and such effect on cell cycle was more marked in CD97siRNA-transfected group (all<0.05).

CONCLUSIONSCD97 plays an important role in the cell growth, proliferation, migration and invasion of breast cancer MDA-MB231 cells, and compared with CD97, CD97may have more effective inhibitory effects on cellular malignant behaviors.

Objective To evaluate patient outcomes of sustained low flow oxygen inhalation therapy with humidifying and non-humidifying oxygen.Methods Randomized controlled trial (RCT) about sustained low flow oxygen therapy with humidifying and non-humidifying oxygen were searched from China National Knowledge Infrastructure (CNKI),Wanfang database,VIP database,PubMed on line.Literatures were searched from the database had been established till now.The primary outcome of the rate of unpleasant respiratory symptoms,oxygen humidification cylinder pollution as well as secondary lower respiratory tract infections were attached importance to.Two reviewers evaluated the literatures independently according to the RCT authentic assessment manual of Cochrane Collaboration.Negotiation took as the measure to attach an agreement.RevMan 5.2 was used to analyses the result of the study.Results A total of 15 met the inclusion criteria,Meta-analysis showed that humidifying and non-humidifying oxygen was not statistically significant with the rate of unpleasant respiratory symptoms (P ＞ 0.05).Non-humidifying group was better than humidity group at the rate of oxygen humidification cylinder pollution and the secondary lower respiratory symptoms (Z=2.50,2.44,P ＜ 0.05).Conclusions Low flow nasal oxygen inhalation therapy may not have to use humidifying oxygen.

Objective To explore a variety of sustainable development under the network environment of doctor-patient communication service platform in the role of reproductive medicine nursing,to improve the reproductive efficiency of medical care services in the new environment,build new doctor-patient communication platform.Methods In reproductive medicine nursing practice,based on the internet to build diversification of doctor-patient communication information service platform,such as reproductive center website,QQ group of doctor-patient communication,Wechat public service platform.Through the questionnaire survey form,understand reproductive center patients through various network platform in the query,the information,make an appointment for reproductive assisted reproduction related information and interact with staff and other aspects of the usage and satisfaction.Results Recycling effective questionnaire was 517 papers.A total of the use of network platform for the overall coverage was more than 80％ of patients,among them the most widely Wechat public platform,utilization rate was 78.1％ (400/512);60.8％ (312/513) patients think Wechat platform using the most convenient.The overall satisfaction rate for center web services platform was 89.5％ (460/514).Conclusions Online platforms provide better nursing efficiency and quality,as well as improving doctor-patient communication based on the internet environment.It provides a new way for sustainable development for nursing system in the field of reproductive medicine.

Objective This research aims to provide a theoretical basis for the nursing care for adolescent malignant lymphoma patients by probing into their emotional experience.Methods In-depth interview of qualitative study is adopted in the interviews with 9 adolescent malignant lymphoma patients,while Colaizzi' s seven-step data analytical approach is utilized to analyze and sort the data before the main idea is concluded and refined.Results These adolescent malignant lymphoma patients,with complicated psychological changes and a suicidal tendency,find it hard to cope with stress.Conclusions Nursing care should be provided to such patients and there should be a respect for their rights to know their disease as well as a timely crisis intervention to those who have a suicidal tendency.Furthermore,due attention should be given to the mental states of the family members and the stability of the family of the patients.We also call for the love and more support for these patients from the government and caring people in society.

Objective To assess the effectiveness of continuous aspiration of subglottic secretions (CASS)in preventing ventilator-associated pneumonia.Methods The relevant randomized controlled trials were searched in the Cochrane Library,Pubmed,CNKI,VIP and Wanfang databases.The experimental group received CASS and the control group did not(NASS group).Two reviewers selected studies according to the inclusion and exclusion criteria,the quality of studies was critically appraised and data were extracted by two reviewers independently.Meta-analysis was conducted by using RevMan 5.2.Results Sixteen randomized controlled trials met the inclusion criteria and enrolled 2 576 patients.According to the meta analysis,compared with NASS,CASS reduced the incidence of ventilator-associated pneumonia,shortened the length of stay in the intensive care unit and the duration of mechanical ventilation.There was no effect on hospital mortality.Conclusions Continuous aspiration of subglottic secretions appears effective in preventing ventilator-associated pneumonia,decreasing the duration of intensive care unit length of stay and mechanical ventilation among critically ill patients with mechanical ventilation.