1.ApoAⅠ and AIBP inhibit P2X7R-mediated pyroptosis in macrophages through ABCA1
Mengjiao CHEN ; Zhenwang ZHAO ; Siqi WANG ; Jianfeng WU ; Dan LIU ; Jin ZOU ; Min ZHANG
Chinese Journal of Arteriosclerosis 2025;33(5):402-411
Aim To explore the effects of apolipoprotein A Ⅰ(ApoA Ⅰ)and apolipoprotein A Ⅰ binding protein(AIBP)on THP-1-derived macrophage pyroptosis.Methods The lactate dehydrogenase(LDH)detection kit was used to evaluate cell membrane integrity,Hoechst33342/PI staining was used to observe cell membrane permeability,ELISA was used to detect the levels of inflammatory factors such as interleukin-1 β(IL-1β)and interleukin-18(IL-18),Western blot was used to detect the expression of pyroptosis-related protein nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3(NLRP3),gasdermin D(GSDMD),cleaved Caspase-1,IL-1β and IL-18.Results Oxidized low density lipoprotein(ox-LDL)upregulated the expression of NLRP3,GSDMD-N,cleaved Caspase-1,IL-1β and IL-18 in THP-1-derived macrophages in a concentration-dependent manner,and promoted the release of IL-1β,IL-18 and LDH(P<0.05 or P<0.01),indicating that ox-LDL induced pyroptosis in THP-1-derived macrophages in a concentration-dependent manner.Co-treatment of macrophages with ApoA Ⅰ and AIBP significantly downregulated the ex-pression of NLRP3,GSDMD-N,cleaved Caspase-1,IL-1β and IL-18,reduced the release of IL-1 β,IL-18 and LDH,and inhibited ox-LDL induced pyroptosis(P<0.05 or P<0.01).After ATP-binding cassette transporter A1(ABCA1)siRNA transfection,co-treatment with ApoA Ⅰ and AIBP had no significant effect on the expression of pyroptosis-related proteins and secretion of inflammatory factors(P>0.05).Co-treatment of macrophages with ApoA Ⅰ and AIBP significantly re-duced the expression of purinergic 2X7R receptor(P2X7R)on the cell membrane,inhibited P2X7R mediated protein ki-nase R(PKR)phosphorylation and NLRP3 inflammasome assembly(P<0.05 or P<0.01).After P2X7R siRNA trans-fection,co-treatment with ApoA Ⅰ and AIBP had no significant effect on the expression of pyroptosis-related proteins and secretion of inflammatory factors(P>0.05).Conclusion ApoA Ⅰ and AIBP reduce the expression of P2X7R on the cell membrane through ABCA1,inhibiting P2X7R/PKR/NLRP3 mediated macrophage pyroptosis.
2.ApoAⅠ and AIBP inhibit P2X7R-mediated pyroptosis in macrophages through ABCA1
Mengjiao CHEN ; Zhenwang ZHAO ; Siqi WANG ; Jianfeng WU ; Dan LIU ; Jin ZOU ; Min ZHANG
Chinese Journal of Arteriosclerosis 2025;33(5):402-411
Aim To explore the effects of apolipoprotein A Ⅰ(ApoA Ⅰ)and apolipoprotein A Ⅰ binding protein(AIBP)on THP-1-derived macrophage pyroptosis.Methods The lactate dehydrogenase(LDH)detection kit was used to evaluate cell membrane integrity,Hoechst33342/PI staining was used to observe cell membrane permeability,ELISA was used to detect the levels of inflammatory factors such as interleukin-1 β(IL-1β)and interleukin-18(IL-18),Western blot was used to detect the expression of pyroptosis-related protein nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3(NLRP3),gasdermin D(GSDMD),cleaved Caspase-1,IL-1β and IL-18.Results Oxidized low density lipoprotein(ox-LDL)upregulated the expression of NLRP3,GSDMD-N,cleaved Caspase-1,IL-1β and IL-18 in THP-1-derived macrophages in a concentration-dependent manner,and promoted the release of IL-1β,IL-18 and LDH(P<0.05 or P<0.01),indicating that ox-LDL induced pyroptosis in THP-1-derived macrophages in a concentration-dependent manner.Co-treatment of macrophages with ApoA Ⅰ and AIBP significantly downregulated the ex-pression of NLRP3,GSDMD-N,cleaved Caspase-1,IL-1β and IL-18,reduced the release of IL-1 β,IL-18 and LDH,and inhibited ox-LDL induced pyroptosis(P<0.05 or P<0.01).After ATP-binding cassette transporter A1(ABCA1)siRNA transfection,co-treatment with ApoA Ⅰ and AIBP had no significant effect on the expression of pyroptosis-related proteins and secretion of inflammatory factors(P>0.05).Co-treatment of macrophages with ApoA Ⅰ and AIBP significantly re-duced the expression of purinergic 2X7R receptor(P2X7R)on the cell membrane,inhibited P2X7R mediated protein ki-nase R(PKR)phosphorylation and NLRP3 inflammasome assembly(P<0.05 or P<0.01).After P2X7R siRNA trans-fection,co-treatment with ApoA Ⅰ and AIBP had no significant effect on the expression of pyroptosis-related proteins and secretion of inflammatory factors(P>0.05).Conclusion ApoA Ⅰ and AIBP reduce the expression of P2X7R on the cell membrane through ABCA1,inhibiting P2X7R/PKR/NLRP3 mediated macrophage pyroptosis.
3.Application of three dimensional printed personalized guide plate assisted arthroscopic ankle arthrodesis in the treatment of ankle arthritis
Guangyi LI ; Cheng WANG ; Jiazheng WANG ; Chenglin WU ; Jieyuan ZHANG ; Jian ZOU ; Jianfeng XUE ; Yan SU ; Guohua MEI ; Zhongmin SHI ; Xin MA
Chinese Journal of Surgery 2024;62(6):572-580
Objective:To compare the efficacy of conventional open ankle fusion and three dimensional(3D) printed guide plate assisted arthroscopic ankle fusion.Methods:A retrospective cohort study was performed on 256 patients with advanced traumatic ankle arthritis, who were admitted to the Department of Orthopaedics, Shanghai Sixth People′s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine from May 2018 to February 2023 and underwent ankle fusion procedures. The study cohort comprised 119 males and 137 females, with an age of (59.6±9.5) years (range: 37 to 83 years). Among them, 175 cases underwent internal fixation with plates and screws (58 cases through the combined medial and lateral approach, and 117 cases through the simple lateral approach), 48 cases underwent internal fixation with screws through the anterior approach (conventional open group), and 33 cases underwent minimally invasive arthroscopic ankle fusion assisted by 3D printed guide plate (3D printed guide plate arthroscopy group). Propensity score matching was employed to achieve a 1∶1 match(caliper value=0.02) between the baseline characteristics of patients in the 3D printed guide plate arthroscopy group and the conventional open group. Perioperative and follow-up data between the two groups were compared using the t-test, Mann-Whitney U test, Wilcoxon signed rank test, χ2 test or corrected χ2 test as appropriate. Results:Matching was successfully achieved with 20 cases in both the 3D printed guide plate arthroscopy group and the conventional open group, and there were no statistically significant differences in baseline characteristics between the two groups (all P>0.05). The operation time in the 3D printed guide plate arthroscopy group was significantly longer than that in the conventional open group ((88.9±5.6) minutes vs. (77.9±11.7) minutes; t=-2.392, P=0.022), while the frequency of intraoperative fluoroscopies ((1.7±0.8) times vs. (5.2±1.2) times; t=10.604, P<0.01) and length of hospitalization ((5.5±0.9) days vs. (6.4±1.5) days; t=2.480, P=0.018) were significantly lower in the 3D printed guide plate arthroscopy group compared to the conventional open group. The fusion rate was 95.0% (19/20) in the 3D printed guide plate arthroscopy group and 85.0% (17/20) in the conventional open group, with no statistically significant difference between the two groups ( χ2=0.278, P=0.598). The fusion time was (12.1±2.0) weeks in the conventional open group and (11.1±1.7) weeks in the 3D printed guide plate arthroscopy group, with no statistically significant difference between the two groups ( t=1.607, P=0.116). At the final follow-up, the American Orthopedic Foot and Ankle Society ankle hindfoot scale was (72.6±5.5)points in the 3D printed guide plate arthroscopy group and (70.5±5.8)points in the conventional open group, with no statistically significant difference between the two groups ( t=-1.003, P=0.322). The pain visual analogue score of the 3D printed guide plate arthroscopy group was ( M(IQR)) 1.50 (1.00) points, lower than that of the conventional open group by 3.00 (1.00) points, with statistically significant differences ( Z=-3.937, P<0.01). There was no significant difference in complication rate between the conventional open group and the 3D printed guide plate arthroscopy group (25.0%(5/20) vs. 5.0%(1/20), χ2=1.765, P=0.184). Conclusion:3D printed guide plate assisted arthroscopic ankle fusion exhibited several advantages, including reduced frequency of fluoroscopies, alleviation of postoperative pain, and decreased complications and length of hospitalization.
4.Association between physical activity and anxiety/depression symptoms among college students in Jiangxi Province
ZHOU Jianfeng, MA Sheng, XIONG Jianping, SONG Yongjing, ZOU Zhiyong
Chinese Journal of School Health 2024;45(12):1722-1726
Objective:
To investigate the association between physical activity frequency and anxiety/depression symptoms among college students in Jiangxi Province, so as to provide a basis for the physical and mental health development of adolescents.
Methods:
From July to October 2023, 5 761 college students from 11 colleges and universities in Jiangxi Province were selected by convenience sampling to conduct an online questionnaire survey regarding physical activity and mental health. Anxiety symptoms and depressive symptoms were assessed by General Anxiety Disorder Scale-7 and Patient Health Questionnaires-9, respectively. Contingency table was used to analyze the distribution of different anxiety and depression symptoms by physical activity frequency, and ordered Logistic regression was used to analyze the association between physical activity frequency with anxiety and depressive symptoms, and stratified analysis was performed based on gender.
Results:
The detection rate of anxiety symptoms of college students was 43.6% ( n =2 513), and the detection rate of depression symptoms was 63.9% ( n =3 682). There were significant differences in the detection rate of anxiety and depression symptoms among different gender and physical activity frequency groups ( χ 2=15.98, 106.32; 30.65, 150.88, P <0.05). The detection rates of severe anxiety and depression symptoms of male and female who never exercise were higher (severe anxiety: 6.1% for male and 5.2% for female, severe depression: 8.7% for male and 7.4% for female).The results of ordered Logistic regression analysis showed that compared with college students who never exercised, male college students who were physically active almost every day were negatively correlated with mild anxiety and moderate depression symptoms ( OR=0.53,0.33,P <0.05). In addition, male college students who do physical activity 3-4 times a week were negatively correlated with moderate depressive symptoms ( OR =0.43), while male college students who do physical activity 1-2 times a week were negatively correlated with severe depressive symptoms ( OR =0.21) ( P <0.05). Physical activity was negatively correlated with different degress of anxiety and depression symptoms in female college students (anxiety: OR =0.27-0.74, depression: OR =0.18-0.75, P < 0.05).
Conclusions
The frequency of physical activity in college students (especially female college students) is negatively correlated with anxiety and depressive symptoms. It is suggested to improve the frequency of physical activity to promote physical and mental health.
5.Application of three dimensional printed personalized guide plate assisted arthroscopic ankle arthrodesis in the treatment of ankle arthritis
Guangyi LI ; Cheng WANG ; Jiazheng WANG ; Chenglin WU ; Jieyuan ZHANG ; Jian ZOU ; Jianfeng XUE ; Yan SU ; Guohua MEI ; Zhongmin SHI ; Xin MA
Chinese Journal of Surgery 2024;62(6):572-580
Objective:To compare the efficacy of conventional open ankle fusion and three dimensional(3D) printed guide plate assisted arthroscopic ankle fusion.Methods:A retrospective cohort study was performed on 256 patients with advanced traumatic ankle arthritis, who were admitted to the Department of Orthopaedics, Shanghai Sixth People′s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine from May 2018 to February 2023 and underwent ankle fusion procedures. The study cohort comprised 119 males and 137 females, with an age of (59.6±9.5) years (range: 37 to 83 years). Among them, 175 cases underwent internal fixation with plates and screws (58 cases through the combined medial and lateral approach, and 117 cases through the simple lateral approach), 48 cases underwent internal fixation with screws through the anterior approach (conventional open group), and 33 cases underwent minimally invasive arthroscopic ankle fusion assisted by 3D printed guide plate (3D printed guide plate arthroscopy group). Propensity score matching was employed to achieve a 1∶1 match(caliper value=0.02) between the baseline characteristics of patients in the 3D printed guide plate arthroscopy group and the conventional open group. Perioperative and follow-up data between the two groups were compared using the t-test, Mann-Whitney U test, Wilcoxon signed rank test, χ2 test or corrected χ2 test as appropriate. Results:Matching was successfully achieved with 20 cases in both the 3D printed guide plate arthroscopy group and the conventional open group, and there were no statistically significant differences in baseline characteristics between the two groups (all P>0.05). The operation time in the 3D printed guide plate arthroscopy group was significantly longer than that in the conventional open group ((88.9±5.6) minutes vs. (77.9±11.7) minutes; t=-2.392, P=0.022), while the frequency of intraoperative fluoroscopies ((1.7±0.8) times vs. (5.2±1.2) times; t=10.604, P<0.01) and length of hospitalization ((5.5±0.9) days vs. (6.4±1.5) days; t=2.480, P=0.018) were significantly lower in the 3D printed guide plate arthroscopy group compared to the conventional open group. The fusion rate was 95.0% (19/20) in the 3D printed guide plate arthroscopy group and 85.0% (17/20) in the conventional open group, with no statistically significant difference between the two groups ( χ2=0.278, P=0.598). The fusion time was (12.1±2.0) weeks in the conventional open group and (11.1±1.7) weeks in the 3D printed guide plate arthroscopy group, with no statistically significant difference between the two groups ( t=1.607, P=0.116). At the final follow-up, the American Orthopedic Foot and Ankle Society ankle hindfoot scale was (72.6±5.5)points in the 3D printed guide plate arthroscopy group and (70.5±5.8)points in the conventional open group, with no statistically significant difference between the two groups ( t=-1.003, P=0.322). The pain visual analogue score of the 3D printed guide plate arthroscopy group was ( M(IQR)) 1.50 (1.00) points, lower than that of the conventional open group by 3.00 (1.00) points, with statistically significant differences ( Z=-3.937, P<0.01). There was no significant difference in complication rate between the conventional open group and the 3D printed guide plate arthroscopy group (25.0%(5/20) vs. 5.0%(1/20), χ2=1.765, P=0.184). Conclusion:3D printed guide plate assisted arthroscopic ankle fusion exhibited several advantages, including reduced frequency of fluoroscopies, alleviation of postoperative pain, and decreased complications and length of hospitalization.
6.Magnetic resonance imaging study on gray matter volume and abnormal functional connectivity in patients with neuropsychiatric systemic lupus erythematosus
Yifan LI ; Tianyi ZHU ; Hongmei ZOU ; Ruiqi QIN ; Jianguo XIA ; Jianfeng HU ; Ji ZHANG ; Weizhong TIAN
Chinese Journal of Neuromedicine 2024;23(11):1120-1128
Objective:To explore the pathogenesis of neuropsychiatric systemic lupus erythematosus (NPSLE) from imaging perspective by analyzing voxel-based morphology (VBM) and functional connectivity (FC) in resting state functional magnetic resonance imaging (rs-fMRI).Methods:Thirty-five patients with NPSLE and 30 patients with non-NPSLE admitted to Department of Rheumatology and Immunology, Taizhou People's Hospital Affiliated to Nanjing Medical University from June 2020 to March 2023 were enrolled; 31 healthy subjects were included as healthy control group during the same period. All subjects completed routine MRI and rs-fMRI, laboratory tests (C3, C4, IgA, IgM and IgG levels), mini-mental state examination (MMSE), hospital anxiety and depression scale (HADS), and fatigue scale for motor and cognitive functions (FSMC). Whole brain gray matter volume in subjects of the 3 groups was analyzed by VBM method, and the brain regions enjoying significant differences in gray matter volume between the NPSLE group and non-NPSLE group were selected as regions of interest (ROIs) for whole brain FC analysis. Partial correlation method was used to analyze the correlations of imaging indexes in brain regions enjoying significant differences with clinical indexes and imaging scores between NPSLE group and non-NPSLE group. Efficacy of imaging indexes in brain regions enjoying significant difference in differentiating NPSLE from non-NPSLE was analyzed by receiver operating characteristic (ROC) curve.Results:(1) Covariance analysis among the 3 groups showed that the gray matter volume in the left inferior frontal gyrus of orbit, left superior frontal gyrus, right rectus gyrus, right transverse temporal gyrus, and right superior frontal gyrus was significantly different among the 3 groups ( P<0.001, FDR corrected); compared with the healthy control group, the NPSLE group had significantly reduced gray matter volume in the left inferior frontal gyrus of orbit, left superior frontal gyrus of orbit, right rectus gyrus, right transverse temporal gyrus, and right superior frontal gyrus ( P<0.001, FDR corrected); compared with the non-NPSLE group, the NPSLE group had significantly decreased gray matter volume in the left inferior frontal gyrus of orbit, right rectus gyrus, and right transverse temporal gyrus ( P<0.001, FDR corrected). (2) Whole brain FC analysis with brain regions enjoying significant differences as seed points showed that Fisher z-transformed FC (zFC) in the right transverse temporal gyrus and bilateral postcentral gyrus of the NPSLE group were significantly decreased ( P<0.001, FDR corrected). (3) Partial correlation analysis showed that, in the NPSLE group, zFC from the right transverse temporal gyrus to left posterior central gyrus was negatively correlated with disease course ( r=-0.390, P=0.027); gray matter volume in the right orbital superior frontal gyrus was negatively correlated with FSMC-cognitive ( r=-0.401, P=0.023); the gray matter volume in the right orbital superior frontal gyrus was negatively correlated with FSMC-motor ( r=-0.374, P=0.035). (4) ROC curve found that gray matter volume in the right rectus gyrus and zFC from the right transverse temporal gyrus to the right posterior central gyrus had relatively high efficacy in differentiating NPSLE from non-NPSLE, with AUC of 0.771 (95% CI: 0.658-0.885, P<0.001) and 0.794 (95% CI: 0.685-0.904, P<0.001), respectively. Conclusion:NPSLE patients have reduced gray matter volume in multiple brain regions (concentrating in the prefrontal limbic system); and reduced FC with some brain regions is noted; multiple indexes are correlated with clinical indexes.
7.A case of Kounis syndrome characterized by cardiac arrest
Hongliang LI ; Lin CHEN ; Mingqiang CHEN ; Feng LIANG ; Jianfeng ZOU
Clinical Medicine of China 2023;39(2):144-146
Kounis syndrome is an acute coronary syndrome associated with mast cell and platelet activation in the case of allergy or allergic injury. This case of Kounis syndrome type Ⅱ patients with myocardial infarction and cardiac arrest is rare. The patient came to the clinic because of wasp sting. During the treatment, he had cardiac arrest. After cardiopulmonary resuscitation, anti allergy, antithrombotic and coronary revascularization, his condition improved. After discharge, he was followed up regularly. After 1 month and 1 year follow-up, the patient had no chest pain, chest tightness, discomfort and allergic reaction.
8.Maslinic acid alleviates myocardial ischemia/reperfusion injury by inhibiting pyroptosis
Lin LI ; Yao SONG ; Gang YANG ; Jianfeng LÜ ; Nianyu HUANG ; Haifeng ZHOU ; Xuejun ZOU
Chinese Journal of Geriatric Heart Brain and Vessel Diseases 2023;25(12):1349-1353
Objective To investigate the mechanism of maslinic acid on pyroptosis and inflammato-ry response in myocardial ischemia/reperfusion(IR)injury.Methods H9C2 cardiomyocytes were randomly divided into control group,control+maslinic acid group,hypoxia reoxygenation(HR)group,and HR+maslinic acid group.Cellular model of HR injury was constructed by hypoxia for 4 h and then reoxygenation for 12 h.Forty-eight male SD rats were randomly divided into Sham group,IR group,IR+maslinic acid group,IR+maslinic acid+Tri group(n=12).Rat model of myocardial IR injury was established by ligating the left anterior descending branch for 30 min followed by reperfusion for 2 h.The viability of cardiomyocytes was detected,the levels of LDH,CK-MB,IL-1β and IL-18 in the supernatant of cardiomyocytes and rat serum samples were detec-ted in each group.Drug-molecular docking was performed to predict the binding site and binding force of maslinic acid and NOD-like receptor thermal protein domain-associated protein 3(NLRP3).Western blotting was used to detect IκBα,NF-κB P65,NLRP3,apoptosis-associated speck-like protein(ASC),and gasdermid D-N terminal(GSDMD-N)in each group of cardiomyo-cytes and myocardial tissues.Results Compared with the Control group,significantly reduced cell viability,enhanced protein levels of p-IκBα,p-NF-κB P65 and higher releases of LDH,IL-1β and IL-18 were observed in the HR group(P<0.05).Maslinic acid treatment reversed HR-induced changes in above indicators(P<0.05).Compared with the Sham group,the protein levels of p-IκBα,p-NF-κB P65,NLRP3,ASC,GSDMD-N and the releases of serum CK-MB,LDH,IL-1βand IL-18 were significantly increased in the IR group(P<0.05).Maslinic acid treatment also reversed above indicators induced by IR injury(P<0.05).The protein levels of p-IκBα,p-NF-κB P65,NLRP3,ASC and GSDMD-N were significantly increased,and the releases of serum CK-MB,LDH,IL-1β and IL-18 were also elevated in the IR+maslinic acid+Tri group than the IR+maslinic acid group(1681.00±136.20 U/L vs 1251.00±213.60 U/L,1776.00±185.80 U/L vs 1330.00±172.50 U/L,4.32±0.45 vs 2.95±0.26,3.89±0.20 vs 2.47±0.29,P<0.05).Conclusion Maslinic acid can show target intervention in NLRP3 activity,thereby inhibiting inflammatory re-sponse and cell pyroptosis,and ultimately attenuate myocardial IR injury effectively.
9.Efficacy of osteochondral fragment fixation using bioabsorbable pins for Hepple Ⅱ osteochondral lesions of the talus in adolescents
Cheng WANG ; Xueqian LI ; Shaoling FU ; Chenglin WU ; Jiazheng WANG ; Jieyuan ZHANG ; Guangyi LI ; Jian ZOU ; Jianfeng XUE ; Yan SU ; Guohua MEI ; Guoxun SONG ; Wenqi GU ; Zhongmin SHI
International Journal of Surgery 2023;50(7):473-480,C2-C3
Objective:To investigate the efficacy of osteochondral fragment fixation using bioabsorbable pins for Hepple Ⅱ osteochondral lesions of the talus (OLT) in adolescents.Methods:Retrospective case analysis was used. The clinical data and follow-up results of 13 adolescent patients (13 feet) with Hepple Ⅱ OLT were all treated with osteochondral fragment fixation using bioabsorbable pins admitted to Shanghai Sixth People′s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine from January 2017 to December 2021 were retrospectively analyzed. There were 7 males and 6 females, with 13 right feet. The age was (14.85±2.23) years old, ranged from 12 to 18 years old. According to the American orthopedic foot and ankle society (AOFAS) ankle-hindfoot score, visual analogue scale (VAS) and SF-36 score before operation and at the last follow-up were used to evaluate the efficacy and function of the patients. Measurement data with normal distribution were represented as mean ± standard deviation( ± s), and the comparison between groups was conducted using the t-test; The mearsurement data with skewness distribution were expressed by M( Q1, Q3), and rank-sum test was used for inter-group comparison. Results:Thirteen adolescent patients (13 feet) with Hepple Ⅱ OLT underwent surgery successfully and were followed up for (25.54±9.95) months. All wounds healed by first intention, and no complications such as wound infection and delayed healing occurred. Preoperative AOFAS ankle-posterior foot score, VAS and SF-36 score were 58.62±3.55, 7.00 (6.50, 8.00) and 68.38±4.81, respectively. At the last follow-up, the scores were 97.38±2.73, 1.00 (0.00, 1.00), 91.15±4.28, respectively, and the results were significantly improved at the last follow-up, with the difference between the two groups statistically significant( P<0.05). Conclusion:Osteochondral fragment fixation using bioabsorbable pins which can promote cartilage repair, significantly improve symptoms, and achieve better clinical satisfaction with fewer complications, is a safe and effective surgical treatment option for Hepple Ⅱ OLT in adolescents with satisfactory short-term clinical outcomes.


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