1.Effectiveness Evaluation of Pre-Service Training for Laboratory Animal Practitioners Based on the "Three-Separation" Model
Fuli WEN ; Yongjun XU ; Yunfeng FU ; Laien XUE ; Linlin CUI ; Junzhu WANG ; Jun LU ; Heping ZHENG
Laboratory Animal and Comparative Medicine 2026;46(5):771-778
ObjectiveTo summarize the application experience of the "Three-Separation" model in the pre-service training of laboratory animal practitioners and scientifically evaluate its training effectiveness. MethodsA retrospective analysis was conducted on 8 009 practitioners who participated in pre-service training at the Fujian Provincial Laboratory Animal Training Base from January 2010 to December 2025. Among them, 1 250 trainees from January 2010 to December 2017 were included in the control group (traditional training model), and 6 759 trainees from January 2018 to December 2025 were included in the evaluation group ("Three-Separation" model, based on separating job requirements, differentiating exam content, and categorizing exam methods). Meanwhile, a questionnaire survey was conducted to evaluate training effectiveness, yielding 1 624 valid questionnaires (585 from the control group and 1 039 from the evaluation group). The "Three-Separation" model simultaneously optimized fee structures to improve affordability and inclusiveness. The comprehensive competencies and training satisfaction of the two groups after training were compared and analyzed using the independent sample t-test, χ² test, and Wilcoxon rank-sum test. ResultsThe overall satisfaction rate of the evaluation group was 92.49% (961/1 039), significantly higher than that of the control group (77.78%, 455/585), with a highly significant difference (χ²=72.50, P<0.001). The evaluation group showed a more favorable distribution of positive, neutral, and negative ratings in dimensions such as course pertinence, time flexibility, assessment rationality, and reasonableness of fees, and the positive rating rates were highly significantly higher than those in the control group (P<0.001). The comprehensive pass rate of the evaluation group (95.53%) was significantly higher than that of the control group (86.24%), with a highly significant difference (P<0.001). ConclusionThe scientific application of the "Three-Separation" model can accurately match the competency requirements of different positions, optimize the training and assessment system, reduce participation barriers by making training fees more affordable and accessible, effectively improve the comprehensive quality and training satisfaction of laboratory animal practitioners, achieve better teaching outcomes, and provide replicable and promotable practical references for the standardized upgrading of the training system in the laboratory animal industry and other industries.
2.Chinese expert consensus on the clinical application of recombinant human thyroid-stimulating hormone (2025 edition)
Chentian SHEN ; Yushen GU ; Hui TAN ; Anren KUANG ; Sijin LI ; Yaming LI ; Jing WANG ; Ningyi JIANG ; Jian TAN ; Zhuoying WANG ; Shaoyan LIU ; Xiaoyi LI ; Jugao FANG ; Dingcun LUO ; Youben FAN ; Hui WANG ; Yuetao WANG ; Xuemei WANG ; Feng WANG ; Wei FU ; Feng SHI ; Xuegong LIU ; Zhongwei LÜ ; Yan XING ; Lin LI ; Hui YANG ; Jun XIN ; Yifan ZHANG ; Yue CHEN ; Yansong LIN ; Wei OUYANG ; Shuzhan YAO ; Changjiu ZHAO ; Xinming ZHAO ; Zairong GAO ; Dingde HUANG ; Qiang WEN ; Xingmin HAN ; Cen LOU ; Weibing MIAO ; Wei FAN ; Quanyong LUO ; Hongcheng SHI
Chinese Journal of Clinical Medicine 2026;33(5):932-944
The recombinant human thyroid-stimulating hormone (rhTSH) can be used for differentiated thyroid cancer (DTC) preparation before 131I therapy, and follow-up. Since its approval by the American Food and Drug Administration (FDA) in 1998, rhTSH has been clinically used in European countries, the United States, and other countries for nearly 30 years. In April 2024, China’s first independently developed rhTSH agent was approved. Based on clinical experience and evidence from domestic and international studies, an expert consensus on the application of rhTSH has been developed following thorough consultations among experts. The consensus offers evidence-based recommendations to guide the appropriate use of rhTSH in clinical practice.
3.The Use of Speech in Screening for Cognitive Decline in Older Adults
Si-Wen WANG ; Xiao-Xiao YIN ; Lin-Lin GAO ; Wen-Jun GUI ; Qiao-Xia HU ; Qiong LOU ; Qin-Wen WANG
Progress in Biochemistry and Biophysics 2025;52(2):456-463
Alzheimer’s disease (AD) is a chronic neurodegenerative disorder that severely affects the health of the elderly, marked by its incurability, high prevalence, and extended latency period. The current approach to AD prevention and treatment emphasizes early detection and intervention, particularly during the pre-AD stage of mild cognitive impairment (MCI), which provides an optimal “window of opportunity” for intervention. Clinical detection methods for MCI, such as cerebrospinal fluid monitoring, genetic testing, and imaging diagnostics, are invasive and costly, limiting their broad clinical application. Speech, as a vital cognitive output, offers a new perspective and tool for computer-assisted analysis and screening of cognitive decline. This is because elderly individuals with cognitive decline exhibit distinct characteristics in semantic and audio information, such as reduced lexical richness, decreased speech coherence and conciseness, and declines in speech rate, voice rhythm, and hesitation rates. The objective presence of these semantic and audio characteristics lays the groundwork for computer-based screening of cognitive decline. Speech information is primarily sourced from databases or collected through tasks involving spontaneous speech, semantic fluency, and reading, followed by analysis using computer models. Spontaneous language tasks include dialogues/interviews, event descriptions, narrative recall, and picture descriptions. Semantic fluency tasks assess controlled retrieval of vocabulary items, requiring participants to extract information at the word level during lexical search. Reading tasks involve participants reading a passage aloud. Summarizing past research, the speech characteristics of the elderly can be divided into two major categories: semantic information and audio information. Semantic information focuses on the meaning of speech across different tasks, highlighting differences in vocabulary and text content in cognitive impairment. Overall, discourse pragmatic disorders in AD can be studied along three dimensions: cohesion, coherence, and conciseness. Cohesion mainly examines the use of vocabulary by participants, with a reduction in the use of nouns, pronouns, verbs, and adjectives in AD patients. Coherence assesses the ability of participants to maintain topics, with a decrease in the number of subordinate clauses in AD patients. Conciseness evaluates the information density of participants, with AD patients producing shorter texts with less information compared to normal elderly individuals. Audio information focuses on acoustic features that are difficult for the human ear to detect. There is a significant degradation in temporal parameters in the later stages of cognitive impairment; AD patients require more time to read the same paragraph, have longer vocalization times, and produce more pauses or silent parts in their spontaneous speech signals compared to normal individuals. Researchers have extracted audio and speech features, developing independent systems for each set of features, achieving an accuracy rate of 82% for both, which increases to 86% when both types of features are combined, demonstrating the advantage of integrating audio and speech information. Currently, deep learning and machine learning are the main methods used for information analysis. The overall diagnostic accuracy rate for AD exceeds 80%, and the diagnostic accuracy rate for MCI also exceeds 80%, indicating significant potential. Deep learning techniques require substantial data support, necessitating future expansion of database scale and continuous algorithm upgrades to transition from laboratory research to practical product implementation.
4.A Fitting Method for Photoacoustic Pump-probe Imaging Based on Phase Correction
Zhuo-Jun XIE ; Hong-Wen ZHONG ; Run-Xiang LIU ; Bo WANG ; Ping XUE ; Bin HE
Progress in Biochemistry and Biophysics 2025;52(2):525-532
ObjectivePhotoacoustic pump-probe imaging can effectively eliminate the interference of blood background signal in traditional photoacoustic imaging, and realize the imaging of weak phosphorescence molecules and their triplet lifetimes in deep tissues. However, background differential noise in photoacoustic pump-probe imaging often leads to large fitting results of phosphorescent molecule concentration and triplet lifetime. Therefore, this paper proposes a novel triplet lifetime fitting method for photoacoustic pump-probe imaging. By extracting the phase of the triplet differential signal and the background noise, the fitting bias caused by the background noise can be effectively corrected. MethodsThe advantages and feasibility of the proposed algorithm are verified by numerical simulation, phantom and in vivo experiments, respectively. ResultsIn the numerical simulation, under the condition of noise intensity being 10% of the signal amplitude, the new method can optimize the fitting deviation from 48.5% to about 5%, and has a higher exclusion coefficient (0.88>0.79), which greatly improves the fitting accuracy. The high specificity imaging ability of photoacoustic pump imaging for phosphorescent molecules has been demonstrated by phantom experiments. In vivo experiments have verified the feasibility of the new fitting method proposed in this paper for fitting phosphoometric lifetime to monitor oxygen partial pressure content during photodynamic therapy of tumors in nude mice. ConclusionThis work will play an important role in promoting the application of photoacoustic pump-probe imaging in biomedicine.
5.Role of Innate Trained Immunity in Diseases
Chuang CHENG ; Yue-Qing WANG ; Xiao-Qin MU ; Xi ZHENG ; Jing HE ; Jun WANG ; Chao TAN ; Xiao-Wen LIU ; Li-Li ZOU
Progress in Biochemistry and Biophysics 2025;52(1):119-132
The innate immune system can be boosted in response to subsequent triggers by pre-exposure to microbes or microbial products, known as “trained immunity”. Compared to classical immune memory, innate trained immunity has several different features. Firstly, the molecules involved in trained immunity differ from those involved in classical immune memory. Innate trained immunity mainly involves innate immune cells (e.g., myeloid immune cells, natural killer cells, innate lymphoid cells) and their effector molecules (e.g., pattern recognition receptor (PRR), various cytokines), as well as some kinds of non-immune cells (e.g., microglial cells). Secondly, the increased responsiveness to secondary stimuli during innate trained immunity is not specific to a particular pathogen, but influences epigenetic reprogramming in the cell through signaling pathways, leading to the sustained changes in genes transcriptional process, which ultimately affects cellular physiology without permanent genetic changes (e.g., mutations or recombination). Finally, innate trained immunity relies on an altered functional state of innate immune cells that could persist for weeks to months after initial stimulus removal. An appropriate inducer could induce trained immunity in innate lymphocytes, such as exogenous stimulants (including vaccines) and endogenous stimulants, which was firstly discovered in bone marrow derived immune cells. However, mature bone marrow derived immune cells are short-lived cells, that may not be able to transmit memory phenotypes to their offspring and provide long-term protection. Therefore, trained immunity is more likely to be relied on long-lived cells, such as epithelial stem cells, mesenchymal stromal cells and non-immune cells such as fibroblasts. Epigenetic reprogramming is one of the key molecular mechanisms that induces trained immunity, including DNA modifications, non-coding RNAs, histone modifications and chromatin remodeling. In addition to epigenetic reprogramming, different cellular metabolic pathways are involved in the regulation of innate trained immunity, including aerobic glycolysis, glutamine catabolism, cholesterol metabolism and fatty acid synthesis, through a series of intracellular cascade responses triggered by the recognition of PRR specific ligands. In the view of evolutionary, trained immunity is beneficial in enhancing protection against secondary infections with an induction in the evolutionary protective process against infections. Therefore, innate trained immunity plays an important role in therapy against diseases such as tumors and infections, which has signature therapeutic effects in these diseases. In organ transplantation, trained immunity has been associated with acute rejection, which prolongs the survival of allografts. However, trained immunity is not always protective but pathological in some cases, and dysregulated trained immunity contributes to the development of inflammatory and autoimmune diseases. Trained immunity provides a novel form of immune memory, but when inappropriately activated, may lead to an attack on tissues, causing autoinflammation. In autoimmune diseases such as rheumatoid arthritis and atherosclerosis, trained immunity may lead to enhance inflammation and tissue lesion in diseased regions. In Alzheimer’s disease and Parkinson’s disease, trained immunity may lead to over-activation of microglial cells, triggering neuroinflammation even nerve injury. This paper summarizes the basis and mechanisms of innate trained immunity, including the different cell types involved, the impacts on diseases and the effects as a therapeutic strategy to provide novel ideas for different diseases.
6.Primary intraosseous synovial sarcoma:a case report and literature review
Wen ZHAO ; Wei-Jun QIAN ; Li LI ; Yan-Min WANG ; Peng-Hui SU ; Chao-Xin ZHANG ; Liang XU ; Tie-Cheng WU ; Jun-Qi LIU ; Ya-Jun WANG
Medical Journal of Chinese People's Liberation Army 2025;50(11):1419-1425
Objective To report a case of tibial synovial sarcoma and review relevant literature to enhance understanding of this disease.Methods The clinical data of a patient with tibial synovial sarcoma treated at Kaifeng Central Hospital were retrospectively analyzed.A literature search was conducted in domestic and international databases,including China National Knowledge Infrastructure(CNKI),Wanfang Data,PubMed,Web of Science,and Embase,up to July 2024.Relevant literature was comprehensively reviewed to summarize the imaging and pathological characteristics,treatment,and prognosis of synovial sarcoma.Results A 29-year-old female patient was admitted with left lower extremity pain.X-ray examination revealed a proximal tibia space-occupying lesion suggestive of malignancy,and a mid-tibial space-occupying lesion considered benign.Contrast-enhanced computed tomography(CT)and plain magnetic resonance imaging(MRI)of the proximal tibial lesion also suggested malignancy.Ultrasound-guided biopsy of the proximal tibial tumor revealed a poorly differentiated malignant tumor.Immunohistochemistry results indicated monophasic synovial sarcoma,requiring genetic testing for definitive diagnosis.The patient underwent wide resection of the proximal left tibial malignancy with tumor-type artificial joint replacement,combined with curettage and bone cement filling for the left mid-tibial lesion under anesthesia.Postoperative pathology of space-occupying lesions in the proximal tibia confirmed monophasic synovial sarcoma,and fluorescence in situ hybridization(FISH)demonstrated a rupture of the synovial sarcoma translocation gene(SYT)(i.e.,SS18 positive).There was no recurrence or metastasis found in the patient during the reexamination 6 months after postoperative chemotherapy.As of July 2024,15 cases of genetically confirmed primary intraosseous synovial sarcoma have been reported internationally.Symptoms included pain and swelling,with a medical history of 1-2 years.The X-ray and CT findings showed osteolytic destruction with bone cortical discontinuity.In 13 cases,the intraosseous masses extended to the extraosseous area;in 2 cases,punctate calcifications were detected within the masses.Plain MRI scan showed iso-signal or hypo-signal on T1WI and hyper-signal,iso-signal,and hypo-signal on fat-suppressed T2WI,and enhanced MRI scan demonstrated heterogeneous enhancement.Pathological examination showed spindle-shaped cells under microscopy.Immunohistochemistry results showed positive epithelial membrane antigen(EMA),broad-spectrum cytokeratin(AE1/AE3),Ewing's sarcoma marker(CD99),and transducin-like enhancer of Split 1(TLE1).Twelve patients underwent surgical treatment;6 patients received adjuvant chemotherapy after surgery,of whom 4 developed local recurrence or distant metastasis at initial diagnosis,and 3 died during follow-up.Among the 6 patients who did not receive adjuvant chemotherapy,3 suffered from recurrence or distant metastasis.Conclusions Primary intraosseous synovial sarcoma is a rare malignant tumor with non-specific clinical manifestations.Imaging features typically include osteolytic destruction and intraosseous masses extending extraosseously,suggesting an intraosseous origin.Pathology and immunohistochemistry aid diagnosis,but definitive confirmation relies on further genetic testing.At present,the main treatment regimens for synovial sarcoma involve comprehensive therapies such as surgery and adjuvant chemotherapy,and the prognosis of patients is poor.
7.Effect of different layers of masseter on the bony structure of facial lateral area
Yu-Qi ZHAO ; Jin-Rui JIANG ; Jin-Ran CHEN ; Ze-Chuan WANG ; Hou-En ZHOU ; Wen-Di XU ; Liu-Jun YONG
Acta Anatomica Sinica 2025;56(2):208-213
Objective To observe the morphology of the superficial,middle,and deep layers of the masseter muscle and related bony structures in the lateral facial region of adults through gross anatomy,and to probe into the effects of these muscle layers on the bony structures of the lateral facial region.Methods The bilateral masseter muscles of 12 adult cadavers were exposed,and the superficial,middle,and deep layers were separated and measured for muscle length,tendon length,and muscle belly length.After the masseter muscles were stripped,the total thickness was measured,and the mandible and zygomatic arch were exposed to measure the angle of the mandibular angle,thickness of the zygomatic arch,and width of the zygomatic arch.Observations were made of the masseter tuberosities,and statistical analysis was conducted on their interrelations.Results The zygomatic arch thickness was positively correlated with the length of superficial,middle and deep masseter muscles and the length of superficial and middle masseter belly(r superficial masseter length=0.624,r middle masseter length=0.787,r deep masseter length=0.423,r superficial masseter belly length=0.493,r middle masseter belly length=0.548).The width of the zygomatic arch was positively correlated with the lengths of the superficial and middle muscle layers and the middle muscle belly length(r superficial masseter length=0.527,r middle masseter length=0.521,r middle masseterbelly length=0.437).The angle of the mandibular angle was only negatively correlated with the middle muscle belly length(r=-0.422).The tuberosities of the superficial and middle masseter muscles were not affected by the corresponding muscle layers;However,the tuberosity of the deep masseter was negatively correlated with the length of the deep muscle and the length of the deep tendon(r deep masseter length=-0.543,r deep masseter tendon length=-0.443).Conclusion In the masseter muscle layers of Chinese individuals,the superficial and middle layers have the most significant impact on the bony structures structures of the lateral facial region.These findings are of guiding significance for the remodeling of structures in the lateral facial region.
8.Effects of estrogen signaling on T cell recruitment and polarization in inflamed skeletal muscle with acute myoinjury
Zi-Wei ZHAO ; Xiao-Ting JIAN ; Jun-Yi XIE ; Jing-Wen HUANG ; Yang-Yang LI ; Qi-Sen WANG ; Zhao-Hong LIAO ; Hua LIAO
Acta Anatomica Sinica 2025;56(6):688-696
Objective To investigate the effects of estrogen signaling on T-cell recruitment and polarization in acutely injured skeletal muscle.Methods One hundred C57BL/6 male mice,one hundred and eighty C57BL/6 female mice were selected.Twenty-five female mice were ovariectomized(OVX)and 10 male mice were taken as the sham-operated(sham).Then,cardiotoxin(CTX)induced tibialis anterior(TA)injury for preparing mice myoinjury model.Subcutaneous injection of 17β-estradiol(E2)or estrogen receptor antagonist 4-hydroxytamoxifen(4-OHT)was performed.A total of 140 mice(70 males and 70 females)were divided into four group including:PBS-male,CTX-male,PBS-female,and CTX-female.Serum estradiol(E2)levels were measured by ELISA,and muscle injury models were validated via HE staining.Subsequently,20 male and 20 female mice were selected for immunofluorescence(IF)and Real-time PCR to assess estrogen receptors(ER)expression in injured muscle tissue.Further,10 male and 40 female mice were allocated into five experimental groups,including CTX,CTX+E2,CTX+4-OHT,CTX+OVX,CTX+sham.HE staining and IF were performed to evaluate inflammatory infiltration in the injured muscle.Additionally,50 female mice were divided into CTX and CTX+OVX groups,and IF combined with flow cytometry were used to analyze T-cell phenotypes and muscle fiber regeneration in the injured muscle.Results In vivo,serum E2 and myofiber ERβ increased post-injury in mice of both sexes,significantly higher in females.Compared to the control group,E2 alleviated inflammation,OVX exacerbated inflammation,increased CD4+T-cell infiltration,elevated T helper 1 cell(Th1)response,decreased regulatory T cells(Tregs),impaired regeneration.In vitro,IFN-γ/LPS significantly upregulated ERβ in myotubes.Conclusion Estrogen signaling critically regulates muscle inflammation.Estrogen deficiency(OVX)delays repair of skeletal muscle by promoting Th1 response and suppressing Tregs function.
9.Effect of left bundle branch area pacing on reducing atrial fibrillation following dual-chamber pacemaker implantation
Fang WANG ; Songhai WEN ; Feng LI ; Jun WU ; Shiyi LONG
Journal of Interventional Radiology 2025;34(8):822-827
Objective To analyze the effects of left bundle branch area pacing(LBBaP)on atrial fibrillation occurrence after dual-chamber pacemaker implantation.Methods The medical records of 87 patients who underwent dual-chamber pacemaker implantation at People's Hospital between May 2021 and September 2022 were retrospectively analyzed.According to different implantation methods,the patients were divided into an observation group(n=45)and a control group(n=42).The control group received traditional right ventricular pacing(RVP),while the observation group received LBBaP.Serum N-terminal pro B-type natriuretic peptide(NT-proBNP),QRS onset-to-end duration(QRSd),echocardiographic parameters,cardiac pacing parameters,complications,atrial high-rate events,and atrial fibrillation incidence were compared between the two groups.Results Twelve months after surgery,serum NT-proBNP levels in the observation group were significantly lower than those in the control group(P<0.05),and QRS duration(QRSd)was shorter in the observation group than in the control group(P<0.05).There were no significant differences in left ventricular ejection fraction(LVEF)or left ventricular end-systolic diameter(LVESD)between the two groups at 12 months postoperatively(P>0.05).Left atrial diameter(LAD)was smaller in the observation group than in the control group at 12 months postoperatively(P<0.05).No significant differences were observed in sensing levels or the proportion of cardiac pacing between the two groups(P>0.05).However,the proportion of ventricular pacing was higher in the observation group than in the control group(P<0.05).Intraoperatively and at 12 months postoperatively,impedance levels were lower in the observation group than in the control group(P<0.05),while threshold values were higher in the observation group(P<0.05).Total complication rates did not differ significantly between the two groups(P>0.05).At 12 months postoperatively,the incidence of atrial fibrillation was similar between the two groups(P>0.05),but the incidence of atrial high-rate events was lower in the observation group than in the control group(P<0.05).At 24 months postoperatively,both the incidence of atrial high-rate events and atrial fibrillation were lower in the observation group than in the control group(P<0.05).Conclusion Compared with traditional RVP implantation,LBBaP implantation can significantly improve left heart function,provide more stable pacing parameters,and reduce the risk of long-term atrial high-rate events and atrial fibrillation.The risk of complications is similar for both groups,but LBBaP implantation increases the pacing threshold.
10.The effect of dronedarone and propafenone in preventing postoperative recurrence in patients with atrial fibrillation and its influence on inflammatory response and myocardial injury
Feng LI ; Jun WU ; Shiyi LONG ; Fang WANG ; Songhai WEN
Journal of Interventional Radiology 2025;34(12):1343-1348
Objective To analyze the effect of dronedarone and propafenone in preventing postoperative recurrence in patients with atrial fibrillation(AF)and to discuss its influence on inflammatory response and myocardial injury.Methods A total of 113 patients with AF,who were admitted to the Qiandongnan People's Hospital of China from July 2022 to March 2024,were enrolled in this study.By using random number table method,the patients were divided into control group(n=56)and study group(n=57).All patients received circumpulmonary vein electrical isolation.After treatment,the patients of control group were given propafenone orally for three months,while the patients of study group received dronedarone orally for three months.Both the preoperative and postoperative 3-month inflammatory response indicators including hypersensitive C-reactive protein(Hs-CRP),leucolipin,nucleotide-bound oligomeric domain-like receptor protein 3(NLRP3)inflammasome,and tumor necrosis factor α(TNF-α),the echocardiographic indicators including left atrial maximum volume(LAMV),left ventricular end-systolic diameter(LVESD),and left ventricular ejection fraction(LVEF),the myocardial injury markers including creatine kinase(CK),cardiac troponin I(cTn I),the central nervous specific protein β(S100β),the creatine kinase isoenzyme(CK-MB),the adverse reactions,and the recurrence of AF were compared between the two groups.Results At 3 months after treatment,the serum levels of hs-CRP,leucolipin,NLRP3 inflammasome and TNF-α were decreased in both groups(all P<0.01),which in the study group were obviously lower than those in the control group(all P<0.001);the levels of LAMV and LVESD were decreased in both groups(both P<0.01),which in the study group were remarkably lower than those in the control group(both P<0.001);the level of LVEF was increased in both groups(P<0.01),which in the study group was significantly higher than that in the control group(P<0.001);the levels of CK,cTn I,S100β,and CK-MB were decreased in both groups(all P<0.01),which in the study group were strikingly lower than those in the control group(all P<0.001).The incidence of adverse drug reactions in control group and in the study group was 5.36%(3/56)and 7.02%(4/57 cases)respectively,and the difference was not statistically significant(P>0.05).Within 3 months after treatment,the recurrence rate in the study group was 5.26%(3/57),which was lower than 19.64%(11/56)in the control group,and the difference was statistically significant(P<0.05).Conclusion Compared with propafenone,the use of dronedarone after circumferential pulmonary vein electrical isolation in patients with AF can improve the cardiac functions,reduce myocardial injury and inflammatory responses,and lower the risk of postoperative recurrence of AF.

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