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 improve the multiple-link operation efficiency, effect and satisfaction of transaxillary dual-plane breast augmentation by optimizing and upgrading the configuration of auxiliary tools.Methods:From January 2019 to May 2021, breast augmentation was performed in 130 female patients (aged 32±8 years) in the Cosmetic and Plastic Center of the First Affiliated Hospital of Harbin Medical University. The study was conducted among 63 patients who were eligible for the criteria of high configuration surgery. 67 patients underwent standard configuration surgery. The average operation time, intraoperative blood loss, drainage volume (24 hours after operation), postoperative visual analog scale (VAS) pain score and satisfaction were statistically analyzed.Results:The data of high configuration method and standard configuration method were compared as follows: average operation time was (78.6±12.2) min / (93.1±12.1) min ( t=15.73, P<0.05); the average intraoperative blood loss was (3.1±1.0) ml / (14.4±3.5) ml ( t=13.83, P<0.05); the drainage volume (24 hours after operation) was (37.2±8.2) ml / (61.4±10.9) ml ( t=20.82, P<0.05); the pain score on the first day after surgery was (6.1±1.7) points / (7.5±1.6) points ( t=8.57, P<0.05). The overall satisfaction rate was 97.1±1.6 / 95.6±2.0 ( t=5.58, P>0.001), at 6 months after operation. No severe complications were found during the follow-up period, such as capsular contracture, hematoma, infection and double bubble deformity. Conclusions:The use of ultrasonic knife with delivery bag is an effective optimization and upgrade of the endoscopic assisted transaxillary dual plane breast augmentation. The advantages of this method are obvious, highly efficient, safe, effective and satisfactory. It is worthy of clinical application and promotion.

Objective To introduce our clinical method of filler injection with the posterior space of temporalis myofascia for temporal hollows,and to discuss the practical practicability,safety and aesthetic outcomes.Methods From Jan.2014 to Jan.2016,female patients with temporal hollows who had underwent the filler injecting into the posterior space of temporalis myofascia were involved in our study.The patients with previous history of surgery or trauma on temporal region were excluded.Hyaluronic acid or autologous fat was selected as filler material based on patients' preference.Then,aesthetic outcomes were followed up.120 cases were selected in this study.Among them,52 cases preferred hyaluronic acid,and the average follow-up time was 7.5 months.68 cases acceped autologous fat grafting with average follow-up for 9.5 months.Results All patients got aesthetic and safe outcome.No serious complications were noted,such as infection,embolism,uneven appearance and so on.The satisfactory rate was high in both patients and surgeons.Conclusions The posterior temporalis myofascia space that is relatively closed,stable,lack of vessels has proved to be the safe plane for temporal injection.Not only our injection method could meet the need for injection procedure safety,but also it could ensure the aesthetic outcomes.In clinical practice,the aesthetic surgeons should be well familiar with the temporal anatomy.

Objective To evaluate the design formulas of the optimal location for splitting pectoralis major in terms of making type Ⅰ or Ⅱ dual-plane implant pocket.Methods Sixty-five patients with micromastia were collected.Breasts were divided into two types according to the soft-tissue pinch thickness of the lower pole:type Ⅰ (thickness＞2 cm;34 cases) and type Ⅱ (thickness ≤2 cm;31 cases).The optimal levels at which the pectoralis major (PM) was severed were 3/4 or 2/4 of the distance of new inframammary fold for type Ⅰ or Ⅱ dual-plane pockets,respectively.All patients completed the pre-and post-operative BREAST-Q augmentation modules before and 6 months after surgery.The scores were changed into hundred-mark system by QScore software.Results The recovery processes were well-off.The breasts contour was good.Patients reported higher scores of satisfaction with breasts,psychosocial well-being and sexual well-being after surgery than before surgery (62.0±8.9 vs 27.9± 4.3,65.0± 17.2 vs 17.4±8.3,60.5± 14.2 vs 30.3±5.5,P＜0.05).The mean satisfaction score for the overall outcome was 90.6±5.4.However,there was no significant difference in physical well-being between before operation and aftre operation (85.3±9.5 vs 84.7± 10.6).No complications such as severe capsular contracture,or displacement occurred.Conclusions The design formulas make the determining procedure of the optimal location for pectoralis major splitting for two types dual-plane implant pockets easier and more exactly.Our modified design method can provide the implant with the optimal soft tissue coverage,and bring desired and stable breast aesthetic outcomes.The higher satisfaction and quality of life reported by patients indicate that the formulas are feasible and worth to recommend.