1.Role of Gold Nanorods Functionalized by Nucleic Acid Nanostructures Carrying Doxorubicin in Synergistic Anti-Cancer Therapy.
Hao WU ; Huang Shui MA ; Xing Han WU ; Qiang SUN ; Lin FENG ; Rui Fang JIANG ; Yan Hong LI ; Quan SHI
Biomedical and Environmental Sciences 2025;38(4):403-415
OBJECTIVE:
Cancer remains a significant global health challenge, necessitating the development of effective treatment approaches. Developing synergistic therapy can provide a highly promising strategy for anti-cancer treatment through combining the benefits of various mechanisms.
METHODS:
In this study, we developed a synergistic strategy for chemo-photothermal therapy by constructing nanocomposites using gold nanorods (GNRs) and tetrahedral framework nucleic acids (tFNA) loaded with the anti-tumor drug doxorubicin (DOX).
RESULTS:
Our in vitro studies have systematically clarified the anti-cancer behaviors of tFNA-DOX@GNR nanocomposites, characterized by their enhanced cellular uptake and proficient lysosomal escape capabilities. It was found that the key role of tFNA-DOX@GNR nanocomposites in tumor ablation is primarily due to their capacity to induce cytotoxicity in tumor cells via a photothermal effect, which generates instantaneous high temperatures. This mechanism introduces various responses in tumor cells, facilitated by the thermal effect and the integrated chemotherapeutic action of DOX. These reactions include the induction of endoplasmic reticulum stress, characterized by elevated reactive oxygen species levels, the promotion of apoptotic cell death, and the suppression of tumor cell proliferation.
CONCLUSION
This work exhibits the potential of synergistic therapy utilizing nanocomposites for cancer treatment and offers a promising avenue for future therapeutic strategies.
Doxorubicin/chemistry*
;
Gold/chemistry*
;
Nanotubes/chemistry*
;
Humans
;
Nanocomposites/chemistry*
;
Cell Line, Tumor
;
Nucleic Acids/chemistry*
;
Antibiotics, Antineoplastic/pharmacology*
;
Antineoplastic Agents/administration & dosage*
2.Construction of a prediction model for muscular invasion in upper urinary tract urothelial carcinoma based on preoperative MRI features
Haonan CHEN ; Lingkai CAI ; Hongyuan DING ; Hao JI ; Tianxiao HONG ; Hao YU ; Qikai WU ; Chaoran ZHAO ; Xiao YANG ; Qiang CAO ; Xiancheng ZHAO ; Pengchao LI ; Qiang LYU
Chinese Journal of Urology 2025;46(9):661-668
Objective:To construct a nomogram based on preoperative MRI imaging features for the prediction of muscle-invasive upper urinary tract urothelial carcinoma(UTUC)and evaluate its performance.Methods:This retrospective cohort study analyzed the clinical data of 99 UTUC patients treated at the First Affiliated Hospital of Nanjing Medical University from April 2018 to May 2024. Among them,69(69.7%)were male and 30(30.3%)were female,with a median age of 67.0 years. All patients underwent preoperative MRI and radical nephroureterectomy. According to postoperative pathology,tumors staged ≥ T 2 were assigned to the muscle-invasive group,and those staged ≤ T 1 were assigned to the non-muscle-invasive group. Baseline data,pathological information,and imaging characteristics were collected and compared between the two groups. Logistic regression analysis was performed to identify risk factors for muscle-invasive UTUC,and a nomogram was constructed. The diagnostic performance of the model was assessed using receiver operating characteristic(ROC)curves,calibration curves,and decision curve analysis(DCA). Results:Among the 99 patients,70(70.7%)were diagnosed with muscle-invasive UTUC,and 29(29.3%)with non-muscle-invasive UTUC. The muscle-invasive group had significantly larger tumor size[4.5(2.8,7.0)cm vs. 3.0(2.3,4.5)cm, P = 0.029],a higher incidence of multifocal tumors[37.1%(26/70)vs. 3.5%(1/29), P < 0.001],patchy tumors[30.0%(21/70)vs. 6.9%(2/29), P = 0.019],spiculated tumor margins[52.9%(37/70)vs. 17.2%(5/29), P = 0.001],tumor compression on renal parenchyma or periureteral/peripelvic fat[68.6%(48/70)vs. 10.3%(3/29), P < 0.001],high-grade pathology[92.9%(65/70)vs. 75.9%(22/29), P = 0.043],lymph node metastasis[28.6%(20/70)vs. 0, P = 0.001],and lymphovascular invasion[42.9%(30/70)vs. 10.3%(3/29), P=0.002]. The apparent diffusion coefficient(ADC)values[0.9(0.8,1.1)× 10 -3 mm2/s vs. 1.1(1.0,1.4)× 10 -3 mm2/s, P < 0.001]and normalized ADC(NADC)values[0.8(0.7,1.0)vs. 0.9(0.8,1.1), P = 0.002]were significantly lower in the muscle-invasive group. Univariate logistic regression identified multifocality,patchy tumor patterns,spiculated tumor margins,tumor compression on renal parenchyma or periureteral/peripelvic fat,and low NADC values as risk factors for muscle-invasive UTUC(all P < 0.05). Multivariate analysis revealed multifocality( OR = 17.903,95% CI 1.650 - 194.253, P = 0.018),tumor compression on renal parenchyma or perirenal / ureteral fat( OR = 14.690,95% CI 3.069 - 70.323, P < 0.001),and low NADC value( OR = 0.016,95% CI 0.001 - 0.471, P = 0.017)as independent risk factors. A nomogram was constructed based on these factors. The area under the ROC curve(AUC)of the model was 0.898(95% CI 0.838 - 0.957),with an optimal cutoff value of 0.639. The model showed an accuracy of 83.8%,sensitivity of 81.4%,and specificity of 89.7%. Calibration curves indicated good calibration,and DCA showed that the model provided substantial clinical net benefit. Conclusions:This study constructed a nomogram based on preoperative MRI features,including tumor multifocality,compression on renal parenchyma or periureteral/peripelvic fat and NADC value,which demonstrates good predictive performances for muscle-invasive UTUC.
3.Diagnosis and treatment guideline for acute cervical spinal cord injury without fracture-dislocation in adults (version 2025)
Qingde WANG ; Tongwei CHU ; Jian DONG ; Liangjie DU ; Haoyu FENG ; Shunwu FAN ; Shiqing FENG ; Yanzheng GAO ; Yong HAI ; Da HE ; Dianming JIANG ; Jianyuan JIANG ; Bin LIN ; Bin LIU ; Baoge LIU ; Fang LI ; Feng LI ; Li LI ; Weishi LI ; Fangcai LI ; Xiaoguang LIU ; Hongjian LIU ; Yong LIU ; Zhongjun LIU ; Shibao LU ; Xuhua LU ; Keya MAO ; Xuexiao MA ; Yong QIU ; Limin RONG ; Jun SHU ; Yueming SONG ; Tiansheng SUN ; Yan WANG ; Zhe WANG ; Zheng WANG ; Bing WANG ; Linfeng WANG ; Yu WANG ; Qinghe WANG ; Jigong WU ; Hong XIA ; Guoyong YIN ; Jinglong YAN ; Wen YUAN ; Yong YANG ; Qiang YANG ; Cao YANG ; Jie ZHAO ; Jianguo ZHANG ; Yue ZHU ; Zezhang ZHU ; Yingjie ZHOU ; Zhongmin ZHANG ; Yan ZENG ; Dingjun HAO ; Baorong HE ; Wei MEI
Chinese Journal of Trauma 2025;41(3):243-252
Cervical spinal cord injury without fracture-dislocation (CSCIWFD) is referred to as a special type of cervical spinal cord injury characterized by traumatic spinal cord dysfunction and no significant bony structural abnormalities on imagines. Duo to the high risk of missed diagnosis during the initial consultation, CSCIWFD may lead to progressive neurological deterioration or even complete paralysis, severely impacting patients′ prognosis. Currently, there are no established consensuses over the diagnosis and treatment of CSCIWFD, such as the lack of evidence-based standards for indications of non-surgical treatment and risk of secondary neurological injury, as well as debates over the optimal timing for surgical intervention and indications for different surgical approaches. To address these issues, the Spine Trauma Group of the Orthopedic Branch of the Chinese Medical Doctor Association organized experts in the relevant fields to formulate Diagnosis and treatment guideline for acute cervical spinal cord injury without fracture- dislocation in adults ( version 2025) . Based on evidence-based medicine and the principles of scientific rigor and clinical applicability, the guidelines proposed 11 recommendations covering terminology, diagnosis, evaluation treatment, and rehabilitation, etc., aiming to standardize the management of CSCIWFD.
4.IsoVISoR: Towards 3D Mesoscale Brain Mapping of Large Mammals at Isotropic Sub-micron Resolution.
Chao-Yu YANG ; Yan SHEN ; Xiaoyang QI ; Lufeng DING ; Yanyang XIAO ; Qingyuan ZHU ; Hao WANG ; Cheng XU ; Pak-Ming LAU ; Pengcheng ZHOU ; Fang XU ; Guo-Qiang BI
Neuroscience Bulletin 2025;41(2):344-348
5.Creation and Exploration of the"Organized Fill-in-the-Blank Format"Disci-pline Construction Model for Forensic Medicine in the New Era
Zhi-Wen WEI ; Hong-Xing WANG ; Jun-Hong SUN ; Hao-Liang FAN ; Hong-Liang SU ; Le-Le WANG ; Wen-Ting HE ; Zhe CHEN ; Jie ZHANG ; Xiang-Jie GUO ; Ji LI ; Geng-Qian ZHANG ; Xin-Hua LIANG ; Jiang-Wei YAN ; Qiang-Qiang ZHANG ; Cai-Rong GAO ; Ying-Yuan WANG ; Hong-Wei WANG ; Jun XIE ; Bo-Feng ZHU ; Ke-Ming YUN
Journal of Forensic Medicine 2025;41(1):25-29
Forensic medicine has been designated as a first-level discipline,presenting new opportunities and challenges for the development of forensic medicine.Since the 1980s,the establishment of foren-sic medicine discipline and the cultivation of high-level forensic talents have become hot topics in the development of forensic medicine in China.Since the 13th Five-Year Plan,the forensic team of Shanxi Medical University has been aiming at the forefront,proposing the development goals of"Five First-class"and the discipline development path"Six Major Achievements".It has selected benchmark disci-plines,identified gaps in disciplinary development,unified thoughts,formulated completion timelines,concentrated superior resources,assigned tasks to individuals,and created an"Organized Fill-in-the-Blank Format"forensic medicine discipline construction model with the characteristics of the new era.The construction model of forensic medicine has achieved good results in the goals,discipline frame-work,scientific research,talent cultivation,discipline team and platform construction,forming a rela-tively complete discipline construction and management system,and accumulating valuable experience for the construction of first-level discipline and high-level talent cultivation of forensic medicine.
6.Early impact of robot-assisted total knee arthroplasty on the treatment of varus knee arthritis.
Xin YANG ; Qing-Hao CHENG ; Fu-Qiang ZHANG ; Hua FAN ; Fu-Kang ZHANG ; Zhuang-Zhuang ZHANG ; Yong-Ze YANG ; An-Ren ZHANG ; Hong-Zhang GUO
China Journal of Orthopaedics and Traumatology 2025;38(4):343-351
OBJECTIVE:
To investigate the clinical efficacy and advantages of robot-assisted total knee arthroplasty (TKA) in patients with varus knee osteoarthritis.
METHODS:
Between October 2022 and June 2023, a total of 59 patients with severe knee osteoarthritis resulting in varus were treated with total knee arthroplasty, aged from 59 to 81 years with an average (70.90±4.63) years, including 19 mals and 40 females. The patients were divided into two groups based on the surgical method used:28 patients in the robot group and 31 patients in the traditional group. The robot group consisted of 8 males and 20 femalse patients, with an average age of (70.54±4.80) years and an average disease duration of (14.89±8.72) months. The traditional group consisted of 11 males and 20 females patients, with an average age of (71.39±4.5) years and an average disease duration of (12.32±6.73) months. The operative duration, amount of bleeding during the operation, postoperative activity time after the operation, hip-knee-ankle angle (HKA), lateral distal femoral angle (LDFA), medial proximal tibial angle (MPTA), and complications were compared between the two groups before and after the operation. Lateral tibia component (LTC), frontal tibia component (FTC), frontal femoral component (FFC) and lateral femoral component (LFC) were measured 6 months after operation Additionally, the degree of knee joint motility, American Knee Society score (KSS), and visual analogue scale(VAS) were compared before and after the operation.
RESULTS:
All patients had gradeⅠwound healing without any complications, and all patients were followed up for 6 to 8 months, with an average of (6.5±1.5) months. There were no significant differences preoperative imaging evaluation indexes (including HKA, LDFA, and MPTA), preoperative knee mobility, preoperative VAS, and preoperative KSS between the two groups (P>0.05). Comparing the operation time (109.11±7.16) min vs. (83.90±7.85) min, length of the incision (16.60±2.33) cm vs. (14.47±1.41) cm, intraoperative bleeding (106.93±6.15) ml vs. (147.97±7.62) ml, postoperative activity time (17.86±1.84) h vs. (21.77±2.68) h, between the two groups showed statistically significant differences (P<0.05). There were significant differences in FFC (88.96±0.84)° vs. (87.93±1.09)° and LFC (88.57±1.10)° vs. (87.16±1.2)° between the two groups at 6 months after operation (P<0.05). The robotic group 1, 3, 6 months after KSS (75.96±3.96), (81.53±3.78), (84.50±3.29) scores, VAS (3.68±0.67), (2.43±0.79), (0.54±0.64), knee joint mobility (113.32±4.72) °, (123.93±3.99) °, (135.36±2.34) °;Traditional group KSS (73.77±4.18), (76.48±3.60), (80.19±3.28) scores, VAS (4.16±1.04), (3.03±0.75), (1.42±0.76) scores, knee joint mobility (109.19±6.95) °, (119.94±6.08) °, (134.48±2.14) °. Compared to before surgery, both groups showed significant improvement in KSS, VAS and knee mobility during the three follow-up visits (P<0.001). Additionally, postoperative HKA (180.39±1.95)° vs. (178.52±2.23)°, LDFA (89.67±0.63) ° vs. (89.63±0.63)°, and MPTA (89.44±0.55)° vs. (89.29±0.60)° were significantly improved in both groups compared to before surgery (P<0.001). The robotic group had higher KSS than the traditional group at 1, 3, and 6 months after surgery (P<0.05). The robotic group also had lower VAS than the traditional group at 1, 3, and 6 months after surgery (P<0.05). Furthermore, knee mobility was higher in the robotic group than those in the traditional group at 1 and 6 months after surgery (P<0.05), but there was no significant difference between the two groups at 6 months after surgery.
CONCLUSION
Robot-assisted total knee arthroplasty is a safe and effective method for total knee replacement. The use of robotics can improve the limb axis and prosthesis alignment for patients with preoperative varus deformity, resulting in better clinical and imaging outcomes compared to the conventional group.
Humans
;
Female
;
Male
;
Arthroplasty, Replacement, Knee/methods*
;
Aged
;
Middle Aged
;
Osteoarthritis, Knee/physiopathology*
;
Aged, 80 and over
;
Robotic Surgical Procedures/methods*
7.Diagnosis and treatment guideline for acute cervical spinal cord injury without fracture-dislocation in adults (version 2025)
Qingde WANG ; Tongwei CHU ; Jian DONG ; Liangjie DU ; Haoyu FENG ; Shunwu FAN ; Shiqing FENG ; Yanzheng GAO ; Yong HAI ; Da HE ; Dianming JIANG ; Jianyuan JIANG ; Bin LIN ; Bin LIU ; Baoge LIU ; Fang LI ; Feng LI ; Li LI ; Weishi LI ; Fangcai LI ; Xiaoguang LIU ; Hongjian LIU ; Yong LIU ; Zhongjun LIU ; Shibao LU ; Xuhua LU ; Keya MAO ; Xuexiao MA ; Yong QIU ; Limin RONG ; Jun SHU ; Yueming SONG ; Tiansheng SUN ; Yan WANG ; Zhe WANG ; Zheng WANG ; Bing WANG ; Linfeng WANG ; Yu WANG ; Qinghe WANG ; Jigong WU ; Hong XIA ; Guoyong YIN ; Jinglong YAN ; Wen YUAN ; Yong YANG ; Qiang YANG ; Cao YANG ; Jie ZHAO ; Jianguo ZHANG ; Yue ZHU ; Zezhang ZHU ; Yingjie ZHOU ; Zhongmin ZHANG ; Yan ZENG ; Dingjun HAO ; Baorong HE ; Wei MEI
Chinese Journal of Trauma 2025;41(3):243-252
Cervical spinal cord injury without fracture-dislocation (CSCIWFD) is referred to as a special type of cervical spinal cord injury characterized by traumatic spinal cord dysfunction and no significant bony structural abnormalities on imagines. Duo to the high risk of missed diagnosis during the initial consultation, CSCIWFD may lead to progressive neurological deterioration or even complete paralysis, severely impacting patients′ prognosis. Currently, there are no established consensuses over the diagnosis and treatment of CSCIWFD, such as the lack of evidence-based standards for indications of non-surgical treatment and risk of secondary neurological injury, as well as debates over the optimal timing for surgical intervention and indications for different surgical approaches. To address these issues, the Spine Trauma Group of the Orthopedic Branch of the Chinese Medical Doctor Association organized experts in the relevant fields to formulate Diagnosis and treatment guideline for acute cervical spinal cord injury without fracture- dislocation in adults ( version 2025) . Based on evidence-based medicine and the principles of scientific rigor and clinical applicability, the guidelines proposed 11 recommendations covering terminology, diagnosis, evaluation treatment, and rehabilitation, etc., aiming to standardize the management of CSCIWFD.
8.Construction of a prediction model for muscular invasion in upper urinary tract urothelial carcinoma based on preoperative MRI features
Haonan CHEN ; Lingkai CAI ; Hongyuan DING ; Hao JI ; Tianxiao HONG ; Hao YU ; Qikai WU ; Chaoran ZHAO ; Xiao YANG ; Qiang CAO ; Xiancheng ZHAO ; Pengchao LI ; Qiang LYU
Chinese Journal of Urology 2025;46(9):661-668
Objective:To construct a nomogram based on preoperative MRI imaging features for the prediction of muscle-invasive upper urinary tract urothelial carcinoma(UTUC)and evaluate its performance.Methods:This retrospective cohort study analyzed the clinical data of 99 UTUC patients treated at the First Affiliated Hospital of Nanjing Medical University from April 2018 to May 2024. Among them,69(69.7%)were male and 30(30.3%)were female,with a median age of 67.0 years. All patients underwent preoperative MRI and radical nephroureterectomy. According to postoperative pathology,tumors staged ≥ T 2 were assigned to the muscle-invasive group,and those staged ≤ T 1 were assigned to the non-muscle-invasive group. Baseline data,pathological information,and imaging characteristics were collected and compared between the two groups. Logistic regression analysis was performed to identify risk factors for muscle-invasive UTUC,and a nomogram was constructed. The diagnostic performance of the model was assessed using receiver operating characteristic(ROC)curves,calibration curves,and decision curve analysis(DCA). Results:Among the 99 patients,70(70.7%)were diagnosed with muscle-invasive UTUC,and 29(29.3%)with non-muscle-invasive UTUC. The muscle-invasive group had significantly larger tumor size[4.5(2.8,7.0)cm vs. 3.0(2.3,4.5)cm, P = 0.029],a higher incidence of multifocal tumors[37.1%(26/70)vs. 3.5%(1/29), P < 0.001],patchy tumors[30.0%(21/70)vs. 6.9%(2/29), P = 0.019],spiculated tumor margins[52.9%(37/70)vs. 17.2%(5/29), P = 0.001],tumor compression on renal parenchyma or periureteral/peripelvic fat[68.6%(48/70)vs. 10.3%(3/29), P < 0.001],high-grade pathology[92.9%(65/70)vs. 75.9%(22/29), P = 0.043],lymph node metastasis[28.6%(20/70)vs. 0, P = 0.001],and lymphovascular invasion[42.9%(30/70)vs. 10.3%(3/29), P=0.002]. The apparent diffusion coefficient(ADC)values[0.9(0.8,1.1)× 10 -3 mm2/s vs. 1.1(1.0,1.4)× 10 -3 mm2/s, P < 0.001]and normalized ADC(NADC)values[0.8(0.7,1.0)vs. 0.9(0.8,1.1), P = 0.002]were significantly lower in the muscle-invasive group. Univariate logistic regression identified multifocality,patchy tumor patterns,spiculated tumor margins,tumor compression on renal parenchyma or periureteral/peripelvic fat,and low NADC values as risk factors for muscle-invasive UTUC(all P < 0.05). Multivariate analysis revealed multifocality( OR = 17.903,95% CI 1.650 - 194.253, P = 0.018),tumor compression on renal parenchyma or perirenal / ureteral fat( OR = 14.690,95% CI 3.069 - 70.323, P < 0.001),and low NADC value( OR = 0.016,95% CI 0.001 - 0.471, P = 0.017)as independent risk factors. A nomogram was constructed based on these factors. The area under the ROC curve(AUC)of the model was 0.898(95% CI 0.838 - 0.957),with an optimal cutoff value of 0.639. The model showed an accuracy of 83.8%,sensitivity of 81.4%,and specificity of 89.7%. Calibration curves indicated good calibration,and DCA showed that the model provided substantial clinical net benefit. Conclusions:This study constructed a nomogram based on preoperative MRI features,including tumor multifocality,compression on renal parenchyma or periureteral/peripelvic fat and NADC value,which demonstrates good predictive performances for muscle-invasive UTUC.
9.Effect of the transcription factor SOX3 on ovarian granulosa cell proliferation and estradiol secretion
Rui CAI ; Hao ZHANG ; Zhuang LIU ; Yuanhua CHEN ; Fenfen XIE ; Qiang HONG
Acta Universitatis Medicinalis Anhui 2024;59(3):371-376,383
Objective To study the effect of sex-determining region Y-frame protein 3(SOX3)on proliferation and estradiol secretion in human ovarian granulosa cells(KGN cell line).Methods The gene sequence of human SOX3(NM_005634.3)was searched in Gene-Bank,an NCBI database,and the target gene SOX3 was amplified by PCR,which was cloned into lentiviral vector pLV-EF1a-GFP-2A-Puro to obtain the overexpression lentiviral re-combinant plasmid pLV-EF1a-GFP-2A-Puro-SOX3;the correctly sequenced overexpressed lentiviral recombinant plasmid as well as packaging plasmids(pGag/Pol,pRev,pVSV-G)were co-transfected into human embryonic kidney cell line(HEK 293T)cells(pLV-SOX3 group),and pLV-EF1a-GFP-2A-Puro and packaging plasmids(pGag/Pol,pRev,pVSV-G)were co-transfected into HEK 293T cells(pLV-NC group),the lentiviral particles of both groups were collected and the titers of the viruses were measured after 48 h of transfection,the lentiviruses of the two groups were infected into KGN cells,and the stably expressed cell lines were obtained after puromycin screening for 2 weeks;real-time fluorescence quantitative PCR(RT-qPCR)and Western blot were used to detect the SOX3 mRNA and protein levels in the two groups;CCK-8 assay was used to detect the proliferative ability of the cells in the two groups;ELISA was used to determine the concentration of estradiol in the two groups.Results The identification of PCR products and sequencing results showed that the SOX3 gene fragment was amplified successfully,and the enzyme digestion and sequencing results indicated that the construction of overexpression lentiviral recombinant plasmid was completed;green fluorescence could be detected after lentiviral infection of HEK 293T cells,which indicated that lentiviral packaging was successful;the lentivirus was screened by puromycin after lentiviral infection of KGN cells,and the cells were observed to express green fluorescence under the fluorescence microscope;RT-qPCR and Western blot assays both showed that the expression level of SOX3 in the pLV-SOX3 group was significantly higher than that in the pLV-NC group(P<0.05).CCK-8 assay results showed that the proliferation ability of the cells in the pLV-SOX3 group significantly increased compared with that in the pLV-NC group(P<0.01).ELISA results showed that estradiol concentration was elevated in the pLV-SOX3 group com-pared with the pLV-NC group(P<0.05).Conclusion Overexpression of the transcription factor SOX3 can pro-mote the proliferation and estradiol secretion of human ovarian granulosa cells KGN.
10.Establishment of a Multiplex Detection Method for Common Bacteria in Blood Based on Human Mannan-Binding Lectin Protein-Conjugated Magnetic Bead Enrichment Combined with Recombinase-Aided PCR Technology
Jin Zi ZHAO ; Ping Xiao CHEN ; Wei Shao HUA ; Yu Feng LI ; Meng ZHAO ; Hao Chen XING ; Jie WANG ; Yu Feng TIAN ; Qing Rui ZHANG ; Na Xiao LYU ; Qiang Zhi HAN ; Xin Yu WANG ; Yi Hong LI ; Xin Xin SHEN ; Jun Xue MA ; Qing Yan TIE
Biomedical and Environmental Sciences 2024;37(4):387-398
Objective Recombinase-aided polymerase chain reaction(RAP)is a sensitive,single-tube,two-stage nucleic acid amplification method.This study aimed to develop an assay that can be used for the early diagnosis of three types of bacteremia caused by Staphylococcus aureus(SA),Pseudomonas aeruginosa(PA),and Acinetobacter baumannii(AB)in the bloodstream based on recombinant human mannan-binding lectin protein(M1 protein)-conjugated magnetic bead(M1 bead)enrichment of pathogens combined with RAP. Methods Recombinant plasmids were used to evaluate the assay sensitivity.Common blood influenza bacteria were used for the specific detection.Simulated and clinical plasma samples were enriched with M1 beads and then subjected to multiple recombinase-aided PCR(M-RAP)and quantitative PCR(qPCR)assays.Kappa analysis was used to evaluate the consistency between the two assays. Results The M-RAP method had sensitivity rates of 1,10,and 1 copies/μL for the detection of SA,PA,and AB plasmids,respectively,without cross-reaction to other bacterial species.The M-RAP assay obtained results for<10 CFU/mL pathogens in the blood within 4 h,with higher sensitivity than qPCR.M-RAP and qPCR for SA,PA,and AB yielded Kappa values of 0.839,0.815,and 0.856,respectively(P<0.05). Conclusion An M-RAP assay for SA,PA,and AB in blood samples utilizing M1 bead enrichment has been developed and can be potentially used for the early detection of bacteremia.


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