1.Establishment and Preliminary Analysis of GP73 Interactome Using Proximity-dependent Labeling Technology
Mu-Yi LIU ; Chang ZHANG ; Meng-Xin YANG ; Xin-Long YAN ; Lu-Ming WAN ; Cong-Wen WEI
Progress in Biochemistry and Biophysics 2026;53(3):711-723
ObjectiveProtein-protein interactions (PPIs) are fundamental to the execution of biological functions within living cells. However, traditional biochemical methods, such as co-immunoprecipitation (Co-IP), often fail to capture transient, weak, or membrane-associated interactions due to the stringent detergent requirements for cell lysis. Proximity labeling (PL) has emerged in recent years as a transformative technology for mapping the proteomes of specific subcellular compartments and identifying dynamic interactomes in situ. Golgi protein 73 (GP73, also known as GOLPH2), a resident type II Golgi transmembrane protein, is a well-recognized clinical biomarker for liver diseases, including hepatocellular carcinoma (HCC). Despite its clinical significance, the comprehensive physiological and pathological functions of GP73 remain partially understood. This study aims to establish an APEX2-mediated proximity labeling system specifically targeting GP73 to map its interactome in a living cellular environment, thereby providing new insights into its molecular roles and regulatory mechanisms. MethodsTo achieve spatial specificity, we first constructed a stable cell line expressing a fusion protein consisting of GP73 and the engineered soybean peroxidase APEX2. The localization of the GP73-APEX2 fusion protein was validated to ensure it correctly targeted the Golgi apparatus. The proximity labeling reaction was initiated by incubating the cells with biotin-phenol (BP) for 30 min, followed by a brief (1 min) treatment with1 mmol/L hydrogen peroxide (H2O2). This catalytic reaction converts BP into highly reactive, short-lived biotin-phenoxyl radicals that covalently attach to endogenous proteins within a small labeling radius of the GP73-APEX2 enzyme. Subsequently, the cells were quenched, and biotinylated proteins were enriched using high-affinity streptavidin-coated magnetic beads. The captured “neighbor” proteins were subjected to on-bead digestion and analyzed via liquid chromatography-tandem mass spectrometry (LC-MS/MS) for high-throughput identification. Rigorous bioinformatics analysis, including Gene Ontology (GO) enrichment, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, and protein-protein interaction network mapping, was performed to interpret the biological significance of the identified candidates. ResultsOur results demonstrate the successful establishment of a robust and sensitive APEX2-based proximity labeling system for GP73. We identified a total of 95 high-confidence interacting proteins that were significantly enriched in the GP73 proximity proteome compared to control groups. Bioinformatics analysis revealed that these interactors were predominantly associated with biological processes such as vesicular transport, protein localization, and, most notably, molecular functions related to “ribosome binding” and “translation regulation”. This suggested an unexpected role for the Golgi-resident GP73 in the cellular translation machinery. To validate these findings, we performed targeted biochemical assays which confirmed a direct interaction between GP73 and the subunits of the eukaryotic translation initiation factor 3 (eIF3) complex, specifically EIF3G and EIF3I. Furthermore, functional validation using the surface sensing of translation (SUnSET) assay—a non-radioactive method to monitor protein synthesis—revealed that the overexpression of GP73 significantly promoted global protein translation levels in the cell, whereas its depletion or inhibition resulted in reduced translation efficiency. ConclusionThis study successfully utilized APEX2-mediated proximity labeling to provide the first systematic map of GP73 interactome in living cells. Our findings uncover a novel, unconventional function of GP73 as a regulator of cellular protein translation, likely mediated through its interaction with the eIF3 complex. This discovery significantly broadens our understanding of the biological roles of GP73 beyond its traditional function in the Golgi apparatus and suggests that it may act as a bridge between Golgi-related trafficking and the protein synthesis machinery. Furthermore, the technical framework established in this study provides a valuable template for investigating other complex organelle-associated protein networks and resolving transient macromolecular interactions in various physiological and pathological contexts.
2.Advances in Clinical Application of Gastric Contrast-Enhanced Ultrasound for Gastric Cancer.
Guan-Mo LIU ; Hua LIANG ; Yang GUI ; Jie LI ; Xin YE ; Wei-Ming KANG
Acta Academiae Medicinae Sinicae 2025;47(5):716-724
Gastric contrast-enhanced ultrasound includes oral contrast-enhanced ultrasound (OCUS) and double contrast-enhanced ultrasound (DCEUS),which can provide valuable clinical information about tumor morphology,vascular characteristics,and treatment responses.OCUS can clearly identify the gastric wall structure and the extent and depth of lesions by applying oral contrast agents.DCEUS,based on OCUS combined with venography,can display the anatomical and perfusion characteristics of lesions.In recent years,gastric contrast agents and imaging techniques have developed rapidly.However,the clinical application of gastric contrast-enhanced ultrasound is still in the developmental stage.This article reviews the clinical status of OCUS and DCEUS in the screening,diagnosis,staging,pathological typing,and treatment evaluation of gastric cancer.Studies have shown that gastric contrast-enhanced ultrasound has high sensitivity and specificity in the assessment of diagnosis and T-staging of gastric cancer.Furthermore,gastric contrast-enhanced ultrasound has the advantages of being cost-effective,convenient,non-invasive,free from radiation exposure,real-time,and easy to repeat.In the diagnosis and treatment of gastric cancer,it is expected to become one of the important imaging assessment tools.
Humans
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Stomach Neoplasms/diagnostic imaging*
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Contrast Media
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Ultrasonography/methods*
3.Exploration of embryonic chromosomal abnormalities in patients with recurrent miscarriage in the population undergoing IVF/ICSI-assisted pregnancy
Ming SHI ; Chen ZHANG ; Xin KANG ; Yuxin WANG ; Yang SHI ; Wenxiu ZHU ; Jing ZHANG
Chinese Journal of Reproduction and Contraception 2025;45(5):489-494
Objective:To investigate the factors associated with chromosomal abnormalities in embryos of patients with recurrent miscarriage in the in vitro fertilization/intracytoplasmic sperm injection and embryo transfer (IVF/ICSI-ET) population, and to establish a prediction model for chromosomal abnormalities. Methods:This was a retrospective case-control study, aborted tissues were collected from 349 patients who attended the Reproductive and Genetic Laboratory Sports New Town Ward of Dalian Women's and Children's Medical Center (Group) after IVF/ICSI from September 2019 to October 2024, and the samples were examined by copy number variation sequencing (CNV-seq) combined with short tandem repeat (STR) technology. According to the test results, the aborted tissues were divided into chromosome normal and chromosome abnormal groups. Factors affecting the occurrence of chromosomal abnormalities were analyzed by univariate analysis and multifactorial logistic regression.Results:1) By CNV-seq combined with STR method, a total of 252 cases (72.21%, 252/349) of chromosomal abnormalities were detected, while 97 cases had normal chromosomes. 2) The results of univariate analysis showed that the differences in female age, female body mass index (BMI), gestational week, number of miscarriages, progesterone level after 14 d post-transplantation, ovarian reserve function, male age, and male BMI were statistically significant between the chromosome normal group and the chromosome abnormal group (all P<0.05). 3) The results of the multifactorial logistic regression model showed that female age ( OR=1.261, 95% CI: 1.137-1.398, P<0.001), female BMI ( OR=1.121, 95% CI: 1.038-1.227, P=0.004), gestational week ( OR=1.406, 95% CI: 1.155-1.711, P=0.001), progesterone level 14 d after transplantation ( OR=1.016, 95% CI: 1.000-1.031, P=0.043), and BMI of the male partner ( OR=1.132, 95% CI: 1.050-1.220, P=0.001) were the independent risk factors of chromosomal abnormalities. 4) There were statistically significant differences in female age, female BMI, gestational week, progesterone level 14 d after transplantation, and male BMI between patients with normal chromosomes and those with trisomy chromosomes in aborted tissues (all P<0.05).Advanced female age was correlated with the occurrence of trisomy 22 ( P<0.05), and there was a correlation between advanced female age and increased male BMI and the occurrence of trisomy 16 (all P<0.05). Conclusion:The increase in maternal age, BMI, gestational age, progesterone levels 14 d after transplantation, and male BMI can all lead to an increase in the rate of chromosomal abnormalities and an increase in the incidence of trisomy. The advanced age of the female, can lead to the occurrence of trisomy 22. The age of the female and the BMI of the male are positively correlated with the abnormality rate of trisomy 16.
4.Mass Spectrometry-based Identification of GP73 Interacting Proteins Reveals Its Regulatory Role on RNA Splicing Efficiency
Chang ZHANG ; Mu-Yi LIU ; Meng-Xin YANG ; Lu-Ming WAN ; Hui ZHONG ; Cong-Wen WEI
Chinese Journal of Biochemistry and Molecular Biology 2025;41(3):404-414
Protein-protein interactions play an extremely important role in the biochemical functions of cells,and in-depth analysis of protein interactions is the key to understanding cellular life activities.In this study,we systematically mined the interacting proteins of Golgi protein 73(GP73)using classical immunoprecipitation combined with mass spectrometry,and sought to further analyze the molecular func-tion of GP73.Hepatocellular carcinoma cell line HepG2 was selected,and a stable cell line overexpress-ing GP73-3Flag was constructed using lentiviral infection technology.A total of 78 high-confidence GP73 interacting proteins were identified by immunoprecipitation coupled with mass spectrometry.Bioinformat-ics analyses suggested that GP73 interacted with nearly 40 cytosolic proteins and participated in the bio-logical processes of RNA transport,splicing,and translation.Further immunofluorescence and cytosolic protein isolation experiments confirmed the cytosolic localization of GP73 in a variety of tumor cells.Based on the 78 interacting proteins,we further screened protein interaction networks related to mRNA splicing and verified the existence of interactions between GP73 and seven proteins,including HNRN-PH3,SMN1,RBM14,andNCBP1,by co-immunoprecipitation experiments.In addition,minigene spli-cing assay results indicated that GP73 inhibited the splicing efficiency of pre-mRNA by cells.This study contributes to the expansion of knowledge regarding the function of GP73 and aids in elucidating its criti-cal role in cell biology and its potential association with diseases.
5.Teaching Practice and Exploration of"Tutorial System"Based on The Cultivation of Scientific Research and Innovation Ability of Medical Students
Qiao ZHANG ; Yin-Feng YANG ; Yue-Li NI ; Zhuo-Ran TENG ; Wen-Jing LIU ; Jing WU ; Yan-Rui WU ; Yu DOU ; Ming HE ; Shu-De LI ; Ping GAN ; Fang YUAN ; Zhe YANG ; Xin-Wang YANG
Chinese Journal of Biochemistry and Molecular Biology 2025;41(3):470-480
The scientific research and innovation capabilities of medical students are intrinsically linked to the sustained and high-quality development of national healthcare initiatives.Cultivating outstanding medi-cal students with independent scientific capabilities and innovative consciousness is a critical component in the education and training of high-level medical professionals.Our investigation revealed that within the imperfections of the cultivating model,some faculty and students at medical schools have an insufficient understanding of scientific research and innovation and lack motivation for engaging in such activities,which hinder the progression of scientific research activities.Consequently,we initiated a teaching practice and exploratory study on the"tutorial system"aimed at fostering medical students'scientific research and innovation abilities.Based on the principle of"research informing teaching,teaching and research advan-cing together,"this study implements a"tutorial system"coordinated by tutors,supplemented by graduate and undergraduate student mentors,to cultivate innovative thinking,stimulate interest in scientific re-search,and enhance practical and research skills among medical students.Through collaborative efforts within"scientific research innovation teams,"various educational methods—including preliminary re-search,in-class and extracurricular activities,intra-group and inter-group interactions,and theoretical and practical applications—are employed to improve and strengthen the cultivation of medical students'scientif-ic research and innovation abilities.This study aims to provide valuable references for optimizing medical education management systems and enhancing the quality of medical student training.
6.Effect of miR-129-3p mimetic on bone loss in tail-suspended mice
Yi WU ; Zi-dong AN ; Yong-jie PANG ; Li-qiang WANG ; Xin-yang WANG ; Yu-hai GAO ; Xue-yan LI ; Ke-ming CHEN
Chinese Pharmacological Bulletin 2025;41(4):703-709
Aim To study whether intravenous injec-tion of miR-129-3p mimetic(agomir)can resist bone loss caused by hind limb disuse,and to provide new i-deas for preventing bone loss in microgravity environ-ment.Methods Forty-eight C57BL/6J male mice were randomly divided into the control group(CON),tail suspension model group(TS),tail suspension+miR-129-3p agomir administration group(miRNA)and tail suspension+miR-129-3p negative sequence agomir control group(NC).The miRNA group was given 4 mg·kg-1 miR-129-3p agomir by intravenous injection into the medial canthus twice a week.The NC agomir group were consistent with those in the miR-129-3p agomir group,and the CON and TS groups were given only equal volumes of normal saline.After four weeks,all mice were sacrificed and samples were collected.Micro-CT scan of femur,three-point femur bending test,serum bone metabolism index detection,oxidative stress index detection and osteogenesis-related protein expression analysis in bone tissue were per-formed.Results After four weeks,the number of tra-becular bone in the TS group was significantly re-duced,and Tb.BMD,Tb.Th,Tb.N,Tb.BS/TV and Tb.BV/TV were significantly lower than those in the CON group(P<0.01).While Tb.Sp TS group was significantly higher than the CON group(P<0.05),the maximum load and flexural strength of the femur significantly decreased(P<0.01),the content of ser-um bone formation index PINP was significantly lower than that of the CON group(P<0.01),and the con-tent of bone resorption index CTX-I was significantly higher than that of the CON group(P<0.01),the content of serum oxidative damage indexes 8-iso-PGF2α and 8-OHdG significantly increased(P<0.01),and the expression of osteogenesis-related pro-teins in bone tissue markedly decreased(P<0.01).However,the increase or decrease of all indexes in miRNA group was significantly lower than that in TS group.Conclusions miR-129-3p mimetic can signifi-cantly reduce bone loss caused by hind limb disuse.This experiment provides a new idea and method for preventing bone loss in microgravity environment.
7.Efficacy analysis of Doxycline in the treatment of Mycoplasma pneumoniae pneumoina in children
Shangwen HOU ; Ming YANG ; Wenjie QI ; Deli XIN
Chinese Journal of Applied Clinical Pediatrics 2025;40(10):766-770
Objective:To investigate the efficacy and safety of Doxycycline in the treatment of Mycoplasma pneumoniae pneumonia (MPP). Methods:Case series study.A retrospective observational study was conducted on 202 patients diagnosed with MPP in Beijing United Family Hospital from January 2022 to January 2024.The patients were divided into 3 groups according to antibiotics used: Azithromycin group which enrolled 93 cases, 60 cases (Azithromycin alone group) were included after excluding 33 cases with adding steroids; Doxycycline group which enrolled 32 case, 31 cases (Doxycycline alone group) were included after excluding 1 case with adding steroids; Azithromycin switch to Doxycycline when diagnosed Macrolides-unresponsive MPP(MUMPP). Azithromycin switch to Doxycycline group which enrolled 77 cases, 73 cases (switch group) were included after excluding 4 cases with adding steroids.Results used the Mann-Whitney U test, Chi-square test or exact test analyse, comparing the differences in defervescene time, hospitalization day, outcomes and side effects in each group. Results:The fever duration in the Doxycycline alone group was 48(36, 48) h, which was significantly shorter than that in the Azithromycin alone group[120(120, 144) h] ( Z=-7.646, P<0.001). The hospitalization time in the Doxycycline alone group was [3(3, 4) d], which was significantly shorter than that in the Azithromycin alone group [4(3, 5) d] ( Z=-3.368, P=0.002). The medium fever duration in the switch group after Azithromycin was changed to Doxycycline was 48 h, which was not statistically significantly different from that in the Doxycycline alone group ( Z=-0.571, P=0.849). The medium hospitalization time in the switch group after Azithromycin changed to Doxycycline was 4 d, which was significantly longer than that in the Doxycycline alone group (3 d) ( Z=-2.759, P=0.008). Among 93 cases enrolled in the Azithromycin group, 33 cases (35.5%) converted to unresponsive MPP(UMPP), 23 cases (24.7%) progressed to refractory MPP (RMPP), and 33 cases (35.5%) added steroids.Among 32 cases included in the Doxycycline group, 1 case (3.1%) converted to UMPP, no case developed to RMPP, and 1 case (3.1%) added steroids.The above-mentioned 3 proportions were significantly different between the two groups (all P<0.001). Among 93 cases enrolled in the Azithromycin group, 11 cases (11.8%) progressed to severe MPP (SMPP), and 13 cases (14.0%) developed complications.In 32 cases included in the Doxycycline group, 1 case (3.1%) progressed to SMPP, and 3 cases (9.4%) developed complications.The difference in these 2 proportions was not statistically significant between the two groups ( P=0.294, 0.760). In 77 patients included in the Azithromycin switch to Doxycycline group, there were 4 cases converting to UMPP, 4 cases converting to RMPP and 4 cases adding steroids, which were not statistically significantly different from those in the Doxycycline group (all P=0.540). In the Azithromycin switch to Doxycycline group, there were 3 cases progressing to SMPP, and 7 cases developed complications, which were not statistically significantly different from those in the Doxycycline group (all P=1.000). Conclusions:Doxycycline can improve the fever symptom, shorten illness duration and hospitalization time, and reduce steroid usage and the proportions of UMPP and RMPP in pediatric MPP.Switching to Doxycycline is recommended for MUMPP patients as Doxycycline is effective for the treatment of pediatric MPP and no tetracycline pigmentation teeth now.
8.Multiple biomarkers risk score for accurately predicting the long-term prognosis of patients with acute coronary syndrome.
Zhi-Yong ZHANG ; Xin-Yu WANG ; Cong-Cong HOU ; Hong-Bin LIU ; Lyu LYU ; Mu-Lei CHEN ; Xiao-Rong XU ; Feng JIANG ; Long LI ; Wei-Ming LI ; Kui-Bao LI ; Juan WANG
Journal of Geriatric Cardiology 2025;22(7):656-667
BACKGROUND:
Biomarkers-based prediction of long-term risk of acute coronary syndrome (ACS) is scarce. We aim to develop a risk score integrating clinical routine information (C) and plasma biomarkers (B) for predicting long-term risk of ACS patients.
METHODS:
We included 2729 ACS patients from the OCEA (Observation of cardiovascular events in ACS patients). The earlier admitted 1910 patients were enrolled as development cohort; and the subsequently admitted 819 subjects were treated as validation cohort. We investigated 10-year risk of cardiovascular (CV) death, myocardial infarction (MI) and all cause death in these patients. Potential variables contributing to risk of clinical events were assessed using Cox regression models and a score was derived using main part of these variables.
RESULTS:
During 16,110 person-years of follow-up, there were 238 CV death/MI in the development cohort. The 7 most important predictors including in the final model were NT-proBNP, D-dimer, GDF-15, peripheral artery disease (PAD), Fibrinogen, ST-segment elevated MI (STEMI), left ventricular ejection fraction (LVEF), termed as CB-ACS score. C-index of the score for predication of cardiovascular events was 0.79 (95% CI: 0.76-0.82) in development cohort and 0.77 (95% CI: 0.76-0.78) in the validation cohort (5832 person-years of follow-up), which outperformed GRACE 2.0 and ABC-ACS risk score. The CB-ACS score was also well calibrated in development and validation cohort (Greenwood-Nam-D'Agostino: P = 0.70 and P = 0.07, respectively).
CONCLUSIONS
CB-ACS risk score provides a useful tool for long-term prediction of CV events in patients with ACS. This model outperforms GRACE 2.0 and ABC-ACS ischemic risk score.
9.Performance assessment of computed tomographic angiography fractional flow reserve using deep learning: SMART trial summary.
Wei ZHANG ; You-Bing YIN ; Zhi-Qiang WANG ; Ying-Xin ZHAO ; Dong-Mei SHI ; Yong-He GUO ; Zhi-Ming ZHOU ; Zhi-Jian WANG ; Shi-Wei YANG ; De-An JIA ; Li-Xia YANG ; Yu-Jie ZHOU
Journal of Geriatric Cardiology 2025;22(9):793-801
BACKGROUND:
Non-invasive computed tomography angiography (CTA)-based fractional flow reserve (CT-FFR) could become a gatekeeper to invasive coronary angiography. Deep learning (DL)-based CT-FFR has shown promise when compared to invasive FFR. To evaluate the performance of a DL-based CT-FFR technique, DeepVessel FFR (DVFFR).
METHODS:
This retrospective study was designed for iScheMia Assessment based on a Retrospective, single-center Trial of CT-FFR (SMART). Patients suspected of stable coronary artery disease (CAD) and undergoing both CTA and invasive FFR examinations were consecutively selected from the Beijing Anzhen Hospital between January 1, 2016 to December 30, 2018. FFR obtained during invasive coronary angiography was used as the reference standard. DVFFR was calculated blindly using a DL-based CT-FFR approach that utilized the complete tree structure of the coronary arteries.
RESULTS:
Three hundred and thirty nine patients (60.5 ±10.0 years and 209 men) and 414 vessels with direct invasive FFR were included in the analysis. At per-vessel level, sensitivity, specificity, accuracy, positive predictive value (PPV) and negative predictive value (NPV) of DVFFR were 94.7%, 88.6%, 90.8%, 82.7%, and 96.7%, respectively. The area under the receiver operating characteristics curve (AUC) was 0.95 for DVFFR and 0.56 for CTA-based assessment with a significant difference (P < 0.0001). At patient level, sensitivity, specificity, accuracy, PPV and NPV of DVFFR were 93.8%, 88.0%, 90.3%, 83.0%, and 95.8%, respectively. The computation for DVFFR was fast with the average time of 22.5 ± 1.9 s.
CONCLUSIONS
The results demonstrate that DVFFR was able to evaluate lesion hemodynamic significance accurately and effectively with improved diagnostic performance over CTA alone. Coronary artery disease (CAD) is a critical disease in which coronary artery luminal narrowing may result in myocardial ischemia. Early and effective assessment of myocardial ischemia is essential for optimal treatment planning so as to improve the quality of life and reduce medical costs.
10.Effectiveness of Acupuncture in Improving Quality of Life for Patients with Advanced Cancer: A Systematic Review and Meta-Analysis.
Xin YU ; Si-Yao GONG ; Qin LUO ; Gui-Xing XU ; Hao TIAN ; Qian LI ; Ming CHEN ; Sha YANG ; Shu-Guang YU
Chinese journal of integrative medicine 2025;31(4):360-371
OBJECTIVE:
To investigate the effect of acupuncture on advanced cancer patients by meta-analysis.
METHODS:
Nine databases (the Cochrane Central Register of Controlled Trials, MEDLINE, Web of Science, Embase, China National Knowledge Infrastructure, the Cumulative Index to Nursing and Allied Health Literature, Chinese Biomedical Literature Database, China Science and Technology Journal Database, and WanFang Data) were searched for randomized controlled trials (RCTs) on acupuncture in advanced cancer patients published from inception to February 13, 2023 and updated to June 1, 2023. Primary outcomes were quality of life (QOL), while secondary outcomes were pain, fatigue, and adverse events (side effects). Data synthesis was performed using RevMan V.5.3 to calculate pooled effect sizes. RoB-2 was used for the risk of bias, and the quality of evidence was assessed using the Grading of Recommendations, Assessment, Development, and Evaluation (GRADE) tool.
RESULTS:
Totally 17 RCTs involving 1,178 participants were included, 15 of which were pooled for meta-analysis. Most studies demonstrated some concern for the overall risk of bias. The pooled data indicated that acupuncture was associated with improved QOL [mean difference (MD)=6.67, 95% confidence interval (CI): 5.09 to 8.26], pain (MD=-1.18, 95% CI -2.28 to -0.08), and adverse events (risk ratio=0.30, 95% CI: 0.26 to 0.57) compared with control groups. Fatigue outcome was not included. Heterogeneity was substantial, and GRADE evidence was very low for both QOL and pain.
CONCLUSIONS
Acupuncture could benefit patients with advanced cancer and is considered safe compared with usual care. However, the evidence regarding QOL and pain outcomes requires further validation. It is crucial to encourage the development of high-quality studies to strengthen this evidence. (Registry No. CRD42023423539).
Humans
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Acupuncture Therapy
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Neoplasms/therapy*
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Quality of Life
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Randomized Controlled Trials as Topic
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Treatment Outcome

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