1.3- to 24-month Follow-up on COVID-19 with Pulmonary Tuberculosis Survivors after Discharge: Results from a Prospective, Multicenter Study
Ya Jing WANG ; Yu Xing ZONG ; Hui Gui WU ; Lin Yuan QI ; Zhen Hui LI ; Yu Xin JI ; Lin TONG ; Lei ZHANG ; Bo Ming YANG ; Ye Pu YANG ; Ke Ji LI ; Rong Fu XIAO ; Song Lin ZHANG ; Hong Yun HU ; De Hong LIU ; Fang Shou XU ; Sheng SUN ; Wei WU ; Ya MAO ; Qing Min LI ; Hua Hao HOU ; Yuan Zhao GONG ; Yang GUO ; Wen Li JIAO ; Jin QIN ; Yi Ding WANG ; Fang WANG ; Li GUAN ; Gang LIN ; Yan MA ; Ping Yan WANG ; Nan Nan SHI
Biomedical and Environmental Sciences 2022;35(12):1091-1099
		                        		
		                        			
		                        			Objective Coronavirus disease 2019 (COVID-19) and tuberculosis (TB) are major public health and social issues worldwide. The long-term follow-up of COVID-19 with pulmonary TB (PTB) survivors after discharge is unclear. This study aimed to comprehensively describe clinical outcomes, including sequela and recurrence at 3, 12, and 24 months after discharge, among COVID-19 with PTB survivors. Methods From January 22, 2020 to May 6, 2022, with a follow-up by August 26, 2022, a prospective, multicenter follow-up study was conducted on COVID-19 with PTB survivors after discharge in 13hospitals from four provinces in China. Clinical outcomes, including sequela, recurrence of COVID-19, and PTB survivors, were collected via telephone and face-to-face interviews at 3, 12, and 24 months after discharge. Results Thirty-two COVID-19 with PTB survivors were included. The median age was 52 (45, 59) years, and 23 (71.9%) were men. Among them, nearly two-thirds (62.5%) of the survivors were moderate, three (9.4%) were severe, and more than half (59.4%) had at least one comorbidity (PTB excluded). The proportion of COVID-19 survivors with at least one sequela symptom decreased from 40.6% at 3 months to 15.8% at 24 months, with anxiety having a higher proportion over a follow-up. Cough and amnesia recovered at the 12-month follow-up, while anxiety, fatigue, and trouble sleeping remained after 24 months. Additionally, one (3.1%) case presented two recurrences of PTB and no re-positive COVID-19 during the follow-up period. Conclusion The proportion of long symptoms in COVID-19 with PTB survivors decreased over time, while nearly one in six still experience persistent symptoms with a higher proportion of anxiety. The recurrence of PTB and the psychological support of COVID-19 with PTB after discharge require more attention.
		                        		
		                        		
		                        		
		                        	
2.Association of Overlapped and Un-overlapped Comorbidities with COVID-19 Severity and Treatment Outcomes: A Retrospective Cohort Study from Nine Provinces in China.
Yan MA ; Dong Shan ZHU ; Ren Bo CHEN ; Nan Nan SHI ; Si Hong LIU ; Yi Pin FAN ; Gui Hui WU ; Pu Ye YANG ; Jiang Feng BAI ; Hong CHEN ; Li Ying CHEN ; Qiao FENG ; Tuan Mao GUO ; Yong HOU ; Gui Fen HU ; Xiao Mei HU ; Yun Hong HU ; Jin HUANG ; Qiu Hua HUANG ; Shao Zhen HUANG ; Liang JI ; Hai Hao JIN ; Xiao LEI ; Chun Yan LI ; Min Qing LI ; Qun Tang LI ; Xian Yong LI ; Hong De LIU ; Jin Ping LIU ; Zhang LIU ; Yu Ting MA ; Ya MAO ; Liu Fen MO ; Hui NA ; Jing Wei WANG ; Fang Li SONG ; Sheng SUN ; Dong Ting WANG ; Ming Xuan WANG ; Xiao Yan WANG ; Yin Zhen WANG ; Yu Dong WANG ; Wei WU ; Lan Ping WU ; Yan Hua XIAO ; Hai Jun XIE ; Hong Ming XU ; Shou Fang XU ; Rui Xia XUE ; Chun YANG ; Kai Jun YANG ; Sheng Li YUAN ; Gong Qi ZHANG ; Jin Bo ZHANG ; Lin Song ZHANG ; Shu Sen ZHAO ; Wan Ying ZHAO ; Kai ZHENG ; Ying Chun ZHOU ; Jun Teng ZHU ; Tian Qing ZHU ; Hua Min ZHANG ; Yan Ping WANG ; Yong Yan WANG
Biomedical and Environmental Sciences 2020;33(12):893-905
		                        		
		                        			Objective:
		                        			Several COVID-19 patients have overlapping comorbidities. The independent role of each component contributing to the risk of COVID-19 is unknown, and how some non-cardiometabolic comorbidities affect the risk of COVID-19 remains unclear.
		                        		
		                        			Methods:
		                        			A retrospective follow-up design was adopted. A total of 1,160 laboratory-confirmed patients were enrolled from nine provinces in China. Data on comorbidities were obtained from the patients' medical records. Multivariable logistic regression models were used to estimate the odds ratio ( 
		                        		
		                        			Results:
		                        			Overall, 158 (13.6%) patients were diagnosed with severe illness and 32 (2.7%) had unfavorable outcomes. Hypertension (2.87, 1.30-6.32), type 2 diabetes (T2DM) (3.57, 2.32-5.49), cardiovascular disease (CVD) (3.78, 1.81-7.89), fatty liver disease (7.53, 1.96-28.96), hyperlipidemia (2.15, 1.26-3.67), other lung diseases (6.00, 3.01-11.96), and electrolyte imbalance (10.40, 3.00-26.10) were independently linked to increased odds of being severely ill. T2DM (6.07, 2.89-12.75), CVD (8.47, 6.03-11.89), and electrolyte imbalance (19.44, 11.47-32.96) were also strong predictors of unfavorable outcomes. Women with comorbidities were more likely to have severe disease on admission (5.46, 3.25-9.19), while men with comorbidities were more likely to have unfavorable treatment outcomes (6.58, 1.46-29.64) within two weeks.
		                        		
		                        			Conclusion
		                        			Besides hypertension, diabetes, and CVD, fatty liver disease, hyperlipidemia, other lung diseases, and electrolyte imbalance were independent risk factors for COVID-19 severity and poor treatment outcome. Women with comorbidities were more likely to have severe disease, while men with comorbidities were more likely to have unfavorable treatment outcomes.
		                        		
		                        		
		                        		
		                        			Adult
		                        			;
		                        		
		                        			Aged
		                        			;
		                        		
		                        			COVID-19/virology*
		                        			;
		                        		
		                        			China/epidemiology*
		                        			;
		                        		
		                        			Comorbidity
		                        			;
		                        		
		                        			Female
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Male
		                        			;
		                        		
		                        			Middle Aged
		                        			;
		                        		
		                        			Retrospective Studies
		                        			;
		                        		
		                        			Severity of Illness Index
		                        			;
		                        		
		                        			Treatment Outcome
		                        			
		                        		
		                        	
3. Analysis of HPLC Fingerprint and Multi-component Chemical Pattern Recognition of Plantaginis Semen from Different Producing Areas in Jiangxi Province
Chao LI ; Huan YU ; Rou WEN ; Li-ping YAN ; Shou-wen ZHANG ; Qian-feng GONG ; Xiao-ping YUAN
Chinese Journal of Experimental Traditional Medical Formulae 2019;25(15):161-167
		                        		
		                        			
		                        			 Objective:To establish a HPLC fingerprint detection method of Plantaginis Semen, and analyze the samples from different producing areas in Jiangxi province by combining with chemical pattern recognition method, and the contents of five ingredients in Plantaginis Semen were determined. Method:A total of 34 batches of Plantaginis Semen medicinal materials were detected by HPLC. The similarity evaluation was carried out by the 2012 edition of similarity evaluation system of chromatographic fingerprint of traditional Chinese medicine. The chromatographic peak information was used as the data source, and three chemical pattern recognition methods were used to comprehensively analyze the quality of this medicinal herb. Quantitative analysis was performed on the 5 active components, including geniposidic acid, plantamajoside, acteoside, galuteolin and isoacteoside. Result:The similarities between Plantaginis Semen samples from different producing areas in Jiangxi province were >0.86. Orthogonal partial least squares-discriminant analysis (OPLS-DA) could distinguish samples from different producing areas, and be used to determine the chemical components, which had strong correlation with the quality of Plantaginis Semen. The contents of 5 active components in samples from different producing areas were different to some degree, especially in the content of plantamajoside. Conclusion:The established HPLC fingerprint of Plantaginis Semen has strong characteristics, combined with chemical pattern recognition method, it can effectively evaluate the quality of Plantaginis Semen and distinguish its producing areas. 
		                        		
		                        		
		                        		
		                        	
4.Alanine solution as enzyme reaction buffer used in A to O blood group conversion.
Su-Bo LI ; Xue ZHANG ; Yin-Ze ZHANG ; Ying-Xia TAN ; Guo-Qiang BAO ; Ying-Li WANG ; Shou-Ping JI ; Feng GONG ; Hong-Wei GAO
Journal of Experimental Hematology 2014;22(3):817-820
		                        		
		                        			
		                        			The aim of this study was to investigate the effect of alanine solution as α-N-acetylgalactosaminidase enzyme reaction buffer on the enzymatic activity of A antigen. The binding ability of α-N-acetylgalactosaminidase with RBC in different reaction buffer such as alanine solution, glycine solution, normal saline (0.9% NaCl), PBS, PCS was detected by Western blot. The results showed that the efficiency of A to O conversion in alanine solution was similar to that in glycine solution, and Western blot confirmed that most of enzymes blinded with RBC in glycine or alanine solution, but few enzymes blinded with RBC in PBS, PCS or normal saline. The evidences indicated that binding of enzyme with RBC was a key element for A to O blood group conversion, while the binding ability of α-N-acetylgalactosaminidase with RBC in alanine or glycine solution was similar. It is concluded that alanine solution can be used as enzyme reaction buffer in A to O blood group conversion. In this buffer, the α-N-acetylgalactosaminidase is closely blinded with RBC and α-N-acetylgalactosaminidase plays efficient enzymatic activity of A antigen.
		                        		
		                        		
		                        		
		                        			ABO Blood-Group System
		                        			;
		                        		
		                        			immunology
		                        			;
		                        		
		                        			Alanine
		                        			;
		                        		
		                        			Blood Grouping and Crossmatching
		                        			;
		                        		
		                        			methods
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Solutions
		                        			;
		                        		
		                        			alpha-N-Acetylgalactosaminidase
		                        			;
		                        		
		                        			immunology
		                        			
		                        		
		                        	
5.Mitochondrial modulation of apoptosis induced by low-dose radiation in mouse testicular cells.
Fang FANG ; Ping Sheng GONG ; Hong Guang ZHAO ; Yu Jing BI ; Gang ZHAO ; Shou Liang GONG ; Zhi Cheng WANG
Biomedical and Environmental Sciences 2013;26(10):820-830
OBJECTIVETo investigate whether apoptosis induced by low-dose radiation (LDR) is regulated by mitochondrial pathways in testicular cells.
METHODSMale mice were exposed to whole-body LDR, and changes in mitochondrial function and in expression of apoptotic factors were analyzed in the testicular cells as follows. Total nitric-oxide synthase (T-NOS) and Na+/K+ ATPase activities were biochemically assayed. Reactive oxygen species (ROS) and mitochondrial membrane potential (Δψm) were determined by flow cytometry using fluorescent probes. Levels of mRNAs encoding cytochrome c (Cyt c) and apoptosis-inducing factor (AIF) were quantified by real-time reverse-transcription PCR (RT-PCR). Expression of Cyt c, AIF, caspase-9, and caspase-3 at the protein level was assessed by western blotting and immunohistochemistry.
RESULTSLDR induced an increase in T-NOS activity and ROS levels, and a decrease in Na+/K+ ATPase activity and mitochondrial Δψm, in the testicular cells. The intensity of these effects increased with time after irradiation and with dose. The cells showed remarkable swelling and vacuolization of mitochondria, and displayed a time- and dose-dependent increase in the expression of Cyt c, AIF, procaspase-9, and procaspase-3. Activation of the two procaspases was confirmed by detection of the cleaved caspases. The changes in expression of the four apoptotic factors were mostly limited to spermatogonia and spermatocytes.
CONCLUSIONLDR can induce testicular cell apoptosis through mitochondrial signaling pathways.
Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspases ; Cytochromes c ; metabolism ; Membrane Potential, Mitochondrial ; drug effects ; Mice ; Mitochondria ; drug effects ; Reactive Oxygen Species ; metabolism
6.Low-dose radiation induces endoplasmic reticulum stress and activates PERK-CHOP signaling pathway in mouse testicular cells.
Fang FANG ; Ping-Sheng GONG ; Xiang-Fu SONG ; Shou-Liang GONG ; Zhi-Cheng WANG
National Journal of Andrology 2012;18(9):777-782
OBJECTIVETo explore the correlation of low-dose radiation with endoplasmic reticulum stress and the activation of the PERK-CHOP signaling pathway in mouse testicular cells.
METHODSHealthy Kunming mice were randomly assigned to time-effect (0, 3, 6, 12 and 24 h of irradiation at 75 mGy) and dose-effect (12 h of irradiation at 0, 50, 75, 100 and 200 mGy) groups. The contents of H202 and MDA were measured by colorimetry with the agent kits, the expressions of GRP78, PERK and CHOP mRNA detected by quantitative RT-PCR, and the levels of GRP7B, PERK, phosphorylated PERK (pho-PERK) and CHOP proteins determined by Western blotting and image analysis.
RESULTSAfter whole-body irradiation of the mice with 75 mGy, the content of H2 02 in the testis tissue was increased with time prolongation, while that of MDA decreased slightly at 3 and 6 h and then increased with the lengthening of time, both increased significantly at 12 and 24 h as compared with those at 0 h (P < 0. 05, P < 0. 01). Apart from reduced levels of GRP78 mRNA at 3 and 24 h and GRP78 protein at 6 h after irradiation, significant increases were found in the mRNA expressions of GRP78 at 12 h, PERK at 3,6, 12 and 24 hand CHOP at 12 and 24 h (P < 0.05, P < 0.01), as well as in the protein levels of GRP78 at 12 and 24 h, pho-PERK at 3, 12 and 24 h and CHOP at 3, 6, 12 and 24 h in comparison with those at 0 h (P < 0. 05, P < 0. 01). No obvious regularity was observed in the change of the PERK protein expression. After 12 h of whole-body irradiation, the content of H202 was increased at 50, 75 and 100 mGy, but decreased slightly at 200 mGy, while that of MDA was increased with dose increasing, with significant increases in the content of H2 02 at 75 and 100 mCy and in that of MDA at 75, 100 and 200 mGy as compared with the 0 mGy group. Apart from the reduced levels of GRP78 mRNA at 50 and 200 mCy, significant increases were found in the mRNA expressions of PERK at 75, 100 and 200 mGy and CHOP at 50, 75, 100 and 200 (P c 0. 05, P < 0.01) as well as in the protein levels of GRP78 at 100 and 200 mGy, pho-PERK at 50, 100 and 200 mGy and CHOP at 50, 75, 100 and 200 mCy as compared with those at 0 mGy (P < 0. 05, P < 0. 01). There were differences in the changes of different protein expressions, but no obvious regularity was seen in the change of the PERK protein expression.
CONCLUSIONLow-dose radiation can induce endoplasmic reticulum stress in mouse testicular cells, and activate the PERK-CHOP signaling pathway.
Animals ; Endoplasmic Reticulum Stress ; radiation effects ; Heat-Shock Proteins ; metabolism ; Male ; Mice ; Mice, Inbred Strains ; Radiation Dosage ; Radiation, Ionizing ; Signal Transduction ; radiation effects ; Testis ; cytology ; metabolism ; radiation effects ; Transcription Factor CHOP ; metabolism ; Whole-Body Irradiation ; eIF-2 Kinase ; metabolism
7.Removal of αGal xenotransplantation antigen by a novel α-galactosidase.
Hong-Wei GAO ; Xue ZHANG ; Su-Bo LI ; Ying-Xia TAN ; Guo-Qiang BAO ; Ying-Li WANG ; Li-Juan XU ; Shou-Ping JI ; Feng GONG
Journal of Experimental Hematology 2012;20(5):1231-1234
		                        		
		                        			
		                        			αGal, a xenotransplantations antigen (XTA), can lead to hyper acute reaction (HAR) in xenotransplantation. α-Galactosidase from B. fragilis is a novel galactosidase belong to CAZy GH110 which can clear the terminal αGal from branched and linear oligosaccharides. This study was purposed to investigate the removal effect of a novel α-galactosidase on α-Gal XTA on surface of red blood cells. The αGal XTA from the red blood cells of cattle, pig, dog and rabbit was digested by using recombinant α-galactosidase; the α-Gal antigens on surface of cells was detected by flow cytometry. The results showed that the XTA was disappeared completely or mainly. It is concluded that the novel α-galactosidase is a potential enzyme to remove the XTA on the surface of xenotransplants and can be used to overcome the HAR in xenotransplantation.
		                        		
		                        		
		                        		
		                        			Animals
		                        			;
		                        		
		                        			Antigens, Heterophile
		                        			;
		                        		
		                        			immunology
		                        			;
		                        		
		                        			Cattle
		                        			;
		                        		
		                        			Dogs
		                        			;
		                        		
		                        			Epitopes
		                        			;
		                        		
		                        			Erythrocytes
		                        			;
		                        		
		                        			immunology
		                        			;
		                        		
		                        			Macaca mulatta
		                        			;
		                        		
		                        			Mice
		                        			;
		                        		
		                        			Mice, Inbred BALB C
		                        			;
		                        		
		                        			Rabbits
		                        			;
		                        		
		                        			Swine
		                        			;
		                        		
		                        			Transplantation, Heterologous
		                        			;
		                        		
		                        			alpha-Galactosidase
		                        			;
		                        		
		                        			immunology
		                        			
		                        		
		                        	
8.A reconstructed B. Fragilis-derived recombinant α-galactosidase developed for human blood type B→O conversion.
Hong-Wei GAO ; Su-Bo LI ; Guo-Qiang BAO ; Ying-Xia TAN ; Ling-Yan WANG ; Si-Hu JIN ; Ying-Li WANG ; Shou-Ping JI ; Feng GONG
Journal of Experimental Hematology 2011;19(2):503-507
		                        		
		                        			
		                        			This study was aimed to prepare a reconstructed B. Fragilis-derived recombinant α-galactosidase developed for human B to O blood group conversion. Based on the construction of recombinant E. Coli (DE3) which can express α-galactosidase, the inducing time and inducer concentration were optimized for high expression of α-galactosidase. Then, the expression products in supernatant were purified by cation and anion exchange column chromatography. The purified α-galactosidase was used to treat B group red blood cells in phosphate buffer (pH 6.8) for 2 hours to prepare O group red blood cells. The results showed that the optimal inducing conditions for α-galactosidase expression were IPTG 0.1 mmol/L, 37°C and 2 hours. The specific enzyme activity of purified protein increased from 0.42 U/mg to 2.1 U/mg as compared with pre-purification. And, the conditions of B to O blood group conversion were 26°C, pH 6.8 (neutral pH condition) and 2 hours. Moreover, 225 µg of the enzyme could converse 1 ml B red blood cells to O completely. It is concluded that the technology of expression and purification of recombinant α-galactosidase has been established, and the purified protein can converse B red blood cells to O completely, which means that an effective enzyme conversing B red blood cells to O has been obtained.
		                        		
		                        		
		                        		
		                        			ABO Blood-Group System
		                        			;
		                        		
		                        			immunology
		                        			;
		                        		
		                        			Bacteroides fragilis
		                        			;
		                        		
		                        			enzymology
		                        			;
		                        		
		                        			Cloning, Molecular
		                        			;
		                        		
		                        			Escherichia coli
		                        			;
		                        		
		                        			metabolism
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Recombinant Proteins
		                        			;
		                        		
		                        			biosynthesis
		                        			;
		                        		
		                        			alpha-Galactosidase
		                        			;
		                        		
		                        			biosynthesis
		                        			
		                        		
		                        	
9.Research progress of new antibacterial drugs that target bacterial quorum sensing systems.
Shou-Liang YIN ; Ya-Jing CHANG ; Su-Ping DENG ; Qing-Chi WANG ; Wen-Gong YU ; Qian-Hong GONG
Acta Pharmaceutica Sinica 2011;46(6):613-621
		                        		
		                        			
		                        			In recent years, antibiotic resistance of bacteria has become a global health crisis. Especially, the new class of "superbug" was found in South Asia, which is resistant to almost known antibiotics and causes worldwide alarm. Through the underlying mechanisms of bacterial pathogenecity, the expression of many pathogen virulence factors is regulated by the process of quorum sensing. Screening efficient quorum sensing inhibitors is an especially compelling approach to the future treatment of bacterial infections and antibiotic resistance. This article focuses on bacterial quorum sensing system, quorum sensing screening model for in vitro and evaluation of animal models in vivo, recent research of quorum sensing inhibitors and so on.
		                        		
		                        		
		                        		
		                        			Animals
		                        			;
		                        		
		                        			Anti-Bacterial Agents
		                        			;
		                        		
		                        			pharmacology
		                        			;
		                        		
		                        			therapeutic use
		                        			;
		                        		
		                        			Bacterial Infections
		                        			;
		                        		
		                        			drug therapy
		                        			;
		                        		
		                        			Disease Models, Animal
		                        			;
		                        		
		                        			Drug Resistance, Bacterial
		                        			;
		                        		
		                        			Drugs, Chinese Herbal
		                        			;
		                        		
		                        			pharmacology
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Pseudomonas aeruginosa
		                        			;
		                        		
		                        			drug effects
		                        			;
		                        		
		                        			pathogenicity
		                        			;
		                        		
		                        			physiology
		                        			;
		                        		
		                        			Quorum Sensing
		                        			;
		                        		
		                        			drug effects
		                        			;
		                        		
		                        			physiology
		                        			;
		                        		
		                        			Virulence
		                        			;
		                        		
		                        			drug effects
		                        			;
		                        		
		                        			Virulence Factors
		                        			;
		                        		
		                        			metabolism
		                        			
		                        		
		                        	
10.Research advance on universal red blood cell engineering.
Ying-Xia TAN ; Shou-Ping JI ; Feng GONG
Journal of Experimental Hematology 2011;19(3):814-819
		                        		
		                        			
		                        			The preparation and application of universal group O donor red blood cells (RBC) are a trend of future transfusion medicine. This article reviewed the technologies for producing universal RBC in recent years. One of them is modification of blood group antigens, which includes two basic methods. One of these two methods is enzymatic cleavage of the terminal immunodominant sugars from carbohydrate chains on the membrane of group A or/and group B RBC, in order to produce so-called enzyme-converted group O (ECO) RBC. ECO RBC have been produced from whole units of B RBC, which then survived normally when given to type A and O individuals in clinical trial. Because of the complexity of group A antigens, conversion of group A RBC (especially A1 RBC) to group O RBC is more difficult. Recently, a new bacterial glycosidase efficiently cleaving antigens on the surface of both A₁ and A₂ RBC has been obtained. Another method is pegylation, which camouflage the antigens on the surface of RBC with non-immunogenic molecules such as polyethylene glycol (PEG) in a non-specific way, to provide O, minor antigen negative phenotype RBC. The second technology is generating universal RBC from stem cells (such as hematopoietic stem cells, human embryonic stem cells) and human dermal fibroblasts, which will provide a new resource for blood supply. Great progress has been made, but a number of challenges still remain for using them in clinical transfusion, including scale-up, effectiveness and safety of prepared RBC. However, these researches will provide solutions for the problems in current transfusion, such as blood supply shortage, blood borne disease and emergency blood transfusion, and enhance the safety of clinical transfusions in the near future.
		                        		
		                        		
		                        		
		                        			ABO Blood-Group System
		                        			;
		                        		
		                        			Cell Culture Techniques
		                        			;
		                        		
		                        			Embryonic Stem Cells
		                        			;
		                        		
		                        			Erythrocyte Count
		                        			;
		                        		
		                        			Erythrocyte Transfusion
		                        			;
		                        		
		                        			Erythrocytes
		                        			;
		                        		
		                        			Hematopoietic Stem Cells
		                        			;
		                        		
		                        			Humans
		                        			
		                        		
		                        	
            
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