Objective:To establish a one-pot lyophilized detection system based on recombinase polymerase amplification (RPA) and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated protein (Cas13a) technology for the rapid diagnosis of cytomegalovirus (CMV) infection in liver transplantation recipients.Methods:This study is a methodology study. CRISPR RNA (crRNA) and RPA primers were designed targeting the CMV gene sequence. Optimal RPA primer sets were screened to establish the RPA-CRISPR/Cas13a-based CMV detection system. The limit of detection (LOD) was evaluated using gradient-diluted CMV plasmid standards. Cross-reactivity was assessed using genomic DNA from common opportunistic viruses in organ transplant recipients. Lyophilized reagents were validated with CMV-negative and positive samples. P-values were computed using two-sample t-tests for pairwise comparisons and one-way ANOVA for multi-group analyses to assess fluorescence value differences. Subsequently, lyophilized reagents were employed to detect 22 plasma samples from liver transplantation recipients collected at Renji Hospital, Shanghai Jiao Tong University School of Medicine, from June 3, 2024, to May 31, 2025. The test results were then compared with those obtained using quantitative real-time polymerase chain reaction (qPCR). Consistency between the two methods was evaluated using the Kappa coefficient calculated by Kappa test.Result:The established RPA-CRISPR/Cas13a system achieved a detection sensitivity of 1 copy/reaction and exhibited no cross-reactivity with other common opportunistic viruses in organ transplantation. Lyophilized RPA-CRISPR/Cas13a reagents demonstrated performance equivalent to non-lyophilized reagents. Concordance between lyophilized reagent detection and qPCR results for 22 clinical samples was 100% (22/22).Conclusion:A lyophilized CMV detection method based on RPA-CRISPR/Cas13a technology was successfully developed and validated for convenient diagnosing CMV infection in liver transplant recipients.

Objective:To establish a one-pot lyophilized detection system based on recombinase polymerase amplification (RPA) and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated protein (Cas13a) technology for the rapid diagnosis of cytomegalovirus (CMV) infection in liver transplantation recipients.Methods:This study is a methodology study. CRISPR RNA (crRNA) and RPA primers were designed targeting the CMV gene sequence. Optimal RPA primer sets were screened to establish the RPA-CRISPR/Cas13a-based CMV detection system. The limit of detection (LOD) was evaluated using gradient-diluted CMV plasmid standards. Cross-reactivity was assessed using genomic DNA from common opportunistic viruses in organ transplant recipients. Lyophilized reagents were validated with CMV-negative and positive samples. P-values were computed using two-sample t-tests for pairwise comparisons and one-way ANOVA for multi-group analyses to assess fluorescence value differences. Subsequently, lyophilized reagents were employed to detect 22 plasma samples from liver transplantation recipients collected at Renji Hospital, Shanghai Jiao Tong University School of Medicine, from June 3, 2024, to May 31, 2025. The test results were then compared with those obtained using quantitative real-time polymerase chain reaction (qPCR). Consistency between the two methods was evaluated using the Kappa coefficient calculated by Kappa test.Result:The established RPA-CRISPR/Cas13a system achieved a detection sensitivity of 1 copy/reaction and exhibited no cross-reactivity with other common opportunistic viruses in organ transplantation. Lyophilized RPA-CRISPR/Cas13a reagents demonstrated performance equivalent to non-lyophilized reagents. Concordance between lyophilized reagent detection and qPCR results for 22 clinical samples was 100% (22/22).Conclusion:A lyophilized CMV detection method based on RPA-CRISPR/Cas13a technology was successfully developed and validated for convenient diagnosing CMV infection in liver transplant recipients.

Objective To investigate risk factors of medication non-adherence in elderly patients with hypertension through Meta-analysis.Methods Literatures on medication non-adherence in elderly patients with hypertension were systematically reviewed across various databases,from inception to September 1st 2023,including CNKI,VIP,Wangfong Database,SinoMed,PubMed,Web of Science,Cochrane Library、EMbase.Meta-analysis was conducted using RevMan 5.4.Results A total of 19 articles were included,comprising a total of 18 220 patients in sample size.Meta analysis identified 10 risk factors,showed significant association(P<0.05)with:age[OR=1.36,95%CI=(1.00,1.86)],place of residence[OR=1.91,95%CI=(1.33,2.75)],method of payment for medical expenses[OR=1.60,95%CI=(1.38,1.86)],income level[OR=1.68,95%CI=(1.45,1.95)],occupational status[OR=1.79,95%CI=(1.37,2.36)],social support[OR=1.43,95%CI=(1.27,1.60)],comorbidity[CI=0.33,95%(0.19,0.58)],frequency of blood pressure measurement[CI=1.53,95%(1.14,2.05)],disease awareness[OR=1.70,95%CI=(1.44,2.00)]and dedication frequency[OR=1.44,95%CI=(1.16,1.79)].In addition,the number of influencing factors such as gender,marital status,depression,cognitive impairment,and complications was included in<3 articles,and only descriptive analysis was conducted.Conclusion Healthcare providers should focus on the identification and management of the risk factors associated with medication non-adherence in elderly patients with hypertension,therefore to reduce the incidence of medication non-adherence.

OBJECTIVE:To compare the composition changes of Aurantii fructus before and after fermentation processing and optimize its fermentation processing technology. METHODS:UPLC was conducted to compare the raw and fermentation processed products of same batch of Aurantii fructus,and ensure the chromatographic peaks after fermentation processing. Using peak areas of 4 chromatographic peaks and mildew characteristics of samples as index,fermentation temperature,humidity and time as factor, L9(34)orthogonal test was designed to optimize the fermentation processing technology,and verified it. RESULTS:After fermenta-tion processing,Aurantii fructus obviously showed 2 new monosaccharide glycosides components;the optimized fermentation tech-nology was as follows as fermentation temperature of 30 ℃,humidity of 70% and time of 7 d;verification test results showed RSD of each indicator of decoction pieces prepared by optimized fermentation technology in 3 tests were lower than 2.0%(n=3). CONCLUSIONS:Fermentation processing may lead obvious chemical composition changes in Aurantii fructus;the optimized fer-mentation processing technology can increase the contents of characteristic peaks.

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