1.Effect of ORF9 gene of porcine circovirus type 2 on PK-15 cells
Mengting BIAN ; Haiying LIANG ; Zhiyong ZENG ; Deyuan TANG ; Bin WANG ; Ni YE ; Jiajia LIU ; Shu HUANG ; Xiangying PAN ; Hongli TIAN
Chinese Journal of Veterinary Science 2024;44(7):1349-1355
		                        		
		                        			
		                        			To investigate the effects of ORF9 gene of porcine circovirus type 2(PCV2)on PK-15,eu-karyotic expression plasmid was constructed and transfected into PK-15 cells,and the effects of overexpression of ORF9 on proliferation,apoptosis and immunization of PK-15 cells were exam-ined by flow cytometry and qRT-PCR.The results showed that ORF9 gene overexpression signifi-cantly up-regulated the expression levels of the ER stress marker gene GRP78,increased the num-ber of S phase cells,accelerated cell cycle progression,increased the apoptosis rate of PK-15 cells,up-regulated the expression levels of apoptosis-related genes caspase-3,caspase-8,caspase-9,p53 and Bax(P<0.01),down-regulated the expression levels of apoptosis-related genes Bcl-2,up-reg-ulated the expression levels of immune-related genes 1L-8,IL-10,NF-κB and TNF-α(P<0.01),and down-regulated the expression levels of immune-related genes IL-2,IFN-β and IL-12(P<0.01).The above results indicate that ORF9 gene may promote the proliferation and apoptosis of PK-15 cells and play a role in the escape process of PK-15 cells.
		                        		
		                        		
		                        		
		                        	
2.Cloning and sequence analysis of the full-length genome of very virulent infectious bursal disease virus
Jiajia LIU ; Haiying LIANG ; Zhiyong ZENG ; Deyuan TANG ; Bin WANG ; Mengting BIAN ; Shu HUANG ; Xiangying PAN ; Hongli TIAN
Chinese Journal of Veterinary Science 2024;44(7):1394-1400,1407
		                        		
		                        			
		                        			In order to understand the genomic characteristics and genetic variation and strain type of infectious bursal disease virus(IBDV)isolate GZGY2022,which caused the death of chickens in Guizhou farm,primers were designed to amplify the whole genome of the isolate,and genetic evo-lution and strain type analysis were performed after cloning and sequencing.The results showed that the A and B segments of IBDV genome were 3 260,2 827 bp,respectively,encoding VP2-VP5 and VP1 genes.The nucleotide sequence homology between the A and B segments of this strain and the VvIBDV were 96.2%-98.7%and 87.7%-98.9%,respectively,which is the highest with NN1172 strain,83.1%-94.7%and 90.1%-91.0%with other strains.The results of genetic evolution and strain type study showed that IBDV strains can be divided into 6 branches according to antigen and virulence,and the A and B segments of the strain were clustered in the evolutionary branch of VvIBDV,and the strain was A3B3 genotype according to the new genotype classification method.The results of amino acid sequence analysis showed that there were 3 and 7 unique amino acid site variations in the A and B segments of the strain,respectively,and 13 unique characteristic amino acid sites in the coding region of the full-length genome were consistent with VvIBDV.The VP2 sequence of segment A has 19 characteristic amino acid identical with VvIBDV,among which hyper variable regions 222A,242I,253Q,256I,279D,284A,294I and 299S were characteristic ami-no acid sites of the VvIBDV,and the heptapeptide region sequence SWSASGS was consistent with the virulent strain.The VP1 sequence of segment B has 10 characteristic amino acid identical with VvIBDV,among which 61I,145T and 287A were the characteristic amino acid sites of the VvIB-DV.In addition,the nucleotide sequence GGTGCC of 777-782 did not form the restriction endo-nuclease site of Kpn Ⅰ,and combined with the triplet site 145/146/147(TEG),the segment B was consistent with the NN1172 strain,showed that its virulence was slightly weaker than that of the B2 strain of VvIBDV.The results of recombination analysis showed that there were no breaks and recombination sites in the sequence of the strain,and no recombination event occurred.In summa-ry,this study found that GZGY2022 strain belonged to the A3B3 genotype non-recombinant VvIB-DV strain,and its special amino acid sites were consistent with the molecular characteristics of VvIBDV.This study lays the foundation for further exploring the genomic characteristics and path-ogenicity of VvIBDV.
		                        		
		                        		
		                        		
		                        	
3. Effect of Gupi Xiaoji Decoction on mitochondrial structure and function of human hepatoma cell line HepG2
Zhuo LIU ; Shu-Xian YU ; Hui-Ying JIAN ; Pu-Hua ZENG ; Zhuo LIU ; Xiao-Ning TAN ; Ke-Xin LI ; Zhen ZHANG ; Zhuo LIU ; Xue-Fei TIAN ; Wen-Hui GAO
Chinese Pharmacological Bulletin 2023;39(4):781-786
		                        		
		                        			
		                        			 Aim To observe the effect of Gupi Xiaoji Decoction (GPXJY) on the structure and function of mitochondria of human hepatoma cell HepG2 cells and explore its possible mechanism. Methods CCK8 was used to detect cell proliferation, Mito-Tracker Green fluorescence staining was used to observe the mitochondrial structure, flow cytometry was used to detect the membrane potential, Elisa was used to detect the ATP content, fluoroscopic electron microscopy was used to observe the microstructure changes, and high-content screening(HCS) was used to detect the related proteins. Results Fluorescence staining showed that GPXJY damaged the mitochondria of HepG2 cells and decreased the content of ATP. The results of flow cytometry showed that GPXJY could reduce the mitochondrial membrane potential of HepG2 cells. The results of electron microscope showed that GPXJY made the mitochondria of cancer cells swell and so on. HCS found that GPXJY significantly reduced the average fluorescence intensity of Bcl-2 in HepG2 cells, and significantly increased the average fluorescence intensity of apoptosis promoting proteins Bax, cytochrome-c, caspase-3 and cleaved-caspase-3, which was statistically significant. Conclusion GPXJY can regulate the structure and function of mitochondria in HepG2 cells. 
		                        		
		                        		
		                        		
		                        	
4.Guideline for postoperative rehabilitation treatment following vertebral augmentation for osteoporotic vertebral compression fracture (version 2022)
Zhengwei XU ; Dingjun HAO ; Liming CHENG ; Baorong HE ; Bohua CHEN ; Chen CHEN ; Fei CHE ; Jian CHEN ; Qixin CHEN ; Liangjie DU ; Shunwu FAN ; Zhong FANG ; Shiqing FENG ; Yanzheng GAO ; Haishan GUAN ; Zhong GUAN ; Hua JIANG ; Weimin JIANG ; Dianming JIANG ; Jun JIANG ; Yue JIANG ; Lijun HE ; Yuan HE ; Bo LI ; Tao LI ; Jianjun LI ; Xigong LI ; Yijian LIANG ; Bin LIN ; Bin LIU ; Bo LIU ; Yong LIU ; Zhibin LIU ; Xuhua LU ; Chao MA ; Lie QIAN ; Renfu QUAN ; Hongxun SANG ; Haibo SHEN ; Jun SHU ; Honghui SUN ; Tiansheng SUN ; Jun TAN ; Mingxing TANG ; Sheng TAO ; Honglin TENG ; Yun TIAN ; Jiwei TIAN ; Qiang WANG ; Xinwei WANG ; Jianhuang WU ; Peigen XIE ; Weihong XU ; Bin YAN ; Yong YANG ; Guoyong YIN ; Xiaobing YU ; Yuhong ZENG ; Guoqing ZHANG ; Xiaobo ZHANG ; Jie ZHAO ; Yue ZHU
Chinese Journal of Trauma 2022;38(11):961-972
		                        		
		                        			
		                        			Osteoporotic vertebral compression fracture (OVCF) can lead to lower back pain and may be even accompanied by scoliosis, neurological dysfunction and other complications, which will affect the daily activities and life quality of patients. Vertebral augmentation is an effective treatment method for OVCF, but it cannot correct unbalance of bone metabolism or improve the osteoporotic status, causing complications like lower back pain, limited spinal activities and vertebral refracture. The post-operative systematic and standardized rehabilitation treatments can improve curative effect and therapeutic efficacy of anti-osteoporosis, reduce risk of vertebral refracture, increase patient compliance and improve quality of life. Since there still lack relevant clinical treatment guidelines for postoperative rehabilitation treatments following vertebral augmentation for OVCF, the current treatments are varied with uneven therapeutic effect. In order to standardize the postoperative rehabilitation treatment, the Spine Trauma Group of the Orthopedic Branch of Chinese Medical Doctor Association organized relevant experts to refer to relevant literature and develop the "Guideline for postoperative rehabilitation treatment following vertebral augmentation for osteoporotic vertebral compression fracture (2022 version)" based on the clinical guidelines published by the American Academy of Orthopedic Surgeons (AAOS) as well as on the principles of scientificity, practicality and advancement. The guideline provided evidence-based recommendations on 10 important issues related to postoperative rehabilitation treatments of OVCF.
		                        		
		                        		
		                        		
		                        	
5.Correlation Between Functional Components in Codonopsis pilosula Roots and Soil Factors
Tian-zeng NIU ; Yu-long WANG ; Qin-wen HOU ; Shu-xian XIAO ; Hong-mei LUO ; A-ke LIU
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(11):164-172
		                        		
		                        			
		                        			ObjectiveTo explore the correlation between the content of 4 functional components in Codonopsis pilosula roots from different areas and soil factors, and thereby to lay a theoretical basis for soil ecological regulation and improvement of quality of C. pilosula roots. MethodThe content of lobetyolin, atractylenolide Ⅲ, alcohol extract, and polysaccharides, as well as soil fertility and 16 soil factors in 24 batches of samples from different producing areas were determined. Principal component analysis (PCA) and Pearson's correlation analysis were used to explore the key soil factors leading to the variation of chemical component content in C. pilosula roots. ResultThe content of lobetyolin and atractylenolide Ⅲ in samples from Longxi was the highest, and the content of polysaccharides peaked in samples from Huguan. The content of lobetyolin was in positive correlation with soil total nitrogen, total phosphorus, total potassium, alkali-hydrolyzable nitrogen, and available potassium (P<0.01), as well as soil organic matter, pH, available manganese, and available zinc (P<0.05). There was a positive correlation between pH and atractylenolide Ⅲ content (P<0.05). Soil total potassium was in positive correlation with alcohol extract and polysaccharide content (P<0.01). Soil available zinc was positively correlated with alcohol extract and the polysaccharide content (P<0.05). Sample sites with higher PCA scores were Pingshun, Huguan, and Longxi, which were significantly positively correlated with the content of polysaccharides in C. pilosula roots in different habitats. ConclusionThe content of functional components in C. pilosula roots can be improved by raising soil organic matter content and applying specific fertilizers. 
		                        		
		                        		
		                        		
		                        	
6.Understanding the phase separation characteristics of nucleocapsid protein provides a new therapeutic opportunity against SARS-CoV-2.
Dan ZHAO ; Weifan XU ; Xiaofan ZHANG ; Xiaoting WANG ; Yiyue GE ; Enming YUAN ; Yuanpeng XIONG ; Shenyang WU ; Shuya LI ; Nian WU ; Tingzhong TIAN ; Xiaolong FENG ; Hantao SHU ; Peng LANG ; Jingxin LI ; Fengcai ZHU ; Xiaokun SHEN ; Haitao LI ; Pilong LI ; Jianyang ZENG
Protein & Cell 2021;12(9):734-740
7.Clinical features of preterm infants with a birth weight less than 1 500 g undergoing different intensities of resuscitation: a multicenter retrospective analysis.
Miao QIAN ; Zhang-Bin YU ; Xiao-Hui CHEN ; Yan XU ; Yue-Lan MA ; Shan-Yu JIANG ; Huai-Yan WANG ; Zeng-Qin WANG ; Liang-Rong HAN ; Shuang-Shuang LI ; Hong-Yan LU ; Jun WAN ; Yan GAO ; Xiao-Qing CHEN ; Li ZHAO ; Ming-Fu WU ; Hong-Juan ZHANG ; Mei XUE ; Ling-Ling ZHU ; Zhao-Fang TIAN ; Wen-Juan TU ; Xin-Ping WU ; Shu-Ping HAN ; Xiao-Qi GU
Chinese Journal of Contemporary Pediatrics 2021;23(6):593-598
		                        		
		                        			OBJECTIVE:
		                        			To evaluate the clinical features of preterm infants with a birth weight less than 1 500 g undergoing different intensities of resuscitation.
		                        		
		                        			METHODS:
		                        			A retrospective analysis was performed for the preterm infants with a birth weight less than 1 500 g and a gestational age less than 32 weeks who were treated in the neonatal intensive care unit of 20 hospitals in Jiangsu, China from January 2018 to December 2019. According to the intensity of resuscitation in the delivery room, the infants were divided into three groups:non-tracheal intubation (
		                        		
		                        			RESULTS:
		                        			Compared with the non-tracheal intubation group, the tracheal intubation and ECPR groups had significantly lower rates of cesarean section and use of antenatal corticosteroid (
		                        		
		                        			CONCLUSIONS
		                        			For preterm infants with a birth weight less than 1 500 g, the higher intensity of resuscitation in the delivery room is related to lower rate of antenatal corticosteroid therapy, lower gestational age, and lower birth weight. The infants undergoing tracheal intubation or ECRP in the delivery room have an increased incidence rate of adverse clinical outcomes. This suggests that it is important to improve the quality of perinatal management and delivery room resuscitation to improve the prognosis of the infants.
		                        		
		                        		
		                        		
		                        			Birth Weight
		                        			;
		                        		
		                        			Cesarean Section
		                        			;
		                        		
		                        			China
		                        			;
		                        		
		                        			Female
		                        			;
		                        		
		                        			Gestational Age
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Infant
		                        			;
		                        		
		                        			Infant, Newborn
		                        			;
		                        		
		                        			Infant, Premature
		                        			;
		                        		
		                        			Pregnancy
		                        			;
		                        		
		                        			Retrospective Studies
		                        			
		                        		
		                        	
8.Delivery room resuscitation and short-term outcomes of extremely preterm and extremely low birth weight infants: a multicenter survey in North China.
Shuai-Jun LI ; Qi FENG ; Xiu-Ying TIAN ; Ying ZHOU ; Yong JI ; Yue-Mei LI ; Shu-Fen ZHAI ; Wei GUO ; Fang ZHANG ; Rong-Xiu ZHENG ; Hai-Ying HE ; Xia LIU ; Jun-Yi WANG ; Hua MEI ; Hong-Yun WANG ; Hua XIE ; Chao-Mei ZENG ; Li MA ; Ping-Ping ZHANG ; Jin-Yu LI ; Xiao-Ying WANG ; Li-Hua LI ; Hong CUI ; Shu-Lan YANG ; Lu CHEN ; Xiao-Hong GU ; Yan-Ju HU ; Sheng-Shun QUE ; Li-Xia SUN ; Ming YANG ; Wen-Li ZHAO ; Qiu-Yan MA ; Hai-Juan WANG ; Jiu-Ye GUO
Chinese Medical Journal 2021;134(13):1561-1568
		                        		
		                        			BACKGROUND:
		                        			Delivery room resuscitation assists preterm infants, especially extremely preterm infants (EPI) and extremely low birth weight infants (ELBWI), in breathing support, while it potentially exerts a negative impact on the lungs and outcomes of preterm infants. This study aimed to assess delivery room resuscitation and discharge outcomes of EPI and ELBWI in China.
		                        		
		                        			METHODS:
		                        			The clinical data of EPI (gestational age [GA] <28 weeks) and ELBWI (birth weight [BW] <1000 g), admitted within 72 h of birth in 33 neonatal intensive care units from five provinces and cities in North China between 2017 and 2018, were analyzed. The primary outcomes were delivery room resuscitation and risk factors for delivery room intubation (DRI). The secondary outcomes were survival rates, incidence of bronchopulmonary dysplasia (BPD), and risk factors for BPD.
		                        		
		                        			RESULTS:
		                        			A cohort of 952 preterm infants were enrolled. The incidence of DRI, chest compressions, and administration of epinephrine was 55.9% (532/952), 12.5% (119/952), and 7.0% (67/952), respectively. Multivariate analysis revealed that the risk factors for DRI were GA <28 weeks (odds ratio [OR], 3.147; 95% confidence interval [CI], 2.082-4.755), BW <1000 g (OR, 2.240; 95% CI, 1.606-3.125), and antepartum infection (OR, 1.429; 95% CI, 1.044-1.956). The survival rate was 65.9% (627/952) and was dependent on GA. The rate of BPD was 29.3% (181/627). Multivariate analysis showed that the risk factors for BPD were male (OR, 1.603; 95% CI, 1.061-2.424), DRI (OR, 2.094; 95% CI, 1.328-3.303), respiratory distress syndrome exposed to ≥2 doses of pulmonary surfactants (PS; OR, 2.700; 95% CI, 1.679-4.343), and mechanical ventilation ≥7 days (OR, 4.358; 95% CI, 2.777-6.837). However, a larger BW (OR, 0.998; 95% CI, 0.996-0.999), antenatal steroid (OR, 0.577; 95% CI, 0.379-0.880), and PS use in the delivery room (OR, 0.273; 95% CI, 0.160-0.467) were preventive factors for BPD (all P < 0.05).
		                        		
		                        			CONCLUSION
		                        			Improving delivery room resuscitation and management of respiratory complications are imperative during early management of the health of EPI and ELBWI.
		                        		
		                        		
		                        		
		                        			Birth Weight
		                        			;
		                        		
		                        			Bronchopulmonary Dysplasia
		                        			;
		                        		
		                        			China/epidemiology*
		                        			;
		                        		
		                        			Delivery Rooms
		                        			;
		                        		
		                        			Female
		                        			;
		                        		
		                        			Gestational Age
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Infant
		                        			;
		                        		
		                        			Infant, Extremely Low Birth Weight
		                        			;
		                        		
		                        			Infant, Extremely Premature
		                        			;
		                        		
		                        			Infant, Newborn
		                        			;
		                        		
		                        			Male
		                        			;
		                        		
		                        			Pregnancy
		                        			
		                        		
		                        	
9.A multicenter survey of antibiotic use in very and extremely low birth weight infants in Hunan Province.
Ming-Jie WANG ; Shao-Jie YUE ; Jin LIN ; Xi-Rong GAO ; Xiao-Ming PENG ; Meng-Yu CHEN ; Hua-Bao PENG ; Bei CAO ; Yun-Qing ZENG ; Shu-Lian WANG ; Bo WEN ; Xi-Lin HUANG ; Xiao-Ping LI ; Ai-Zhen ZHANG ; Ting CAO ; Yi-Hua CHEN ; Tie-Qiang CHEN ; Chun-Hua YE ; Tao BO ; De-Lin JIANG ; Xiu-Qun HUANG ; Na-Fang REN ; Long-Zhang TAO ; Fang YAO ; Chang-Jun TIAN ; Hong-Ming LI ; Ai-Min ZHANG ; Fu-Rong HUANG ; Wei-Guo ZHANG ; Xiang-Hong CHEN ; Yu-Chan LIU ; Zheng-Lin LIU ; Yan-Shan XU ; Jing-Song MING ; Li CHEN ; Ning-Yi ZHU ; Jun-Min HE ; Sai-Jun YI ; Tuan-Mei WANG ; Zhao-Hui LI ; Gui-Tian WANG
Chinese Journal of Contemporary Pediatrics 2020;22(6):561-566
		                        		
		                        			OBJECTIVE:
		                        			To investigate the current status of antibiotic use for very and extremely low birth weight (VLBW/ELBW) infants in neonatal intensive care units (NICUs) of Hunan Province.
		                        		
		                        			METHODS:
		                        			The use of antibiotics was investigated in multiple level 3 NICUs of Hunan Province for VLBW and ELBW infants born between January, 2017 and December, 2017.
		                        		
		                        			RESULTS:
		                        			The clinical data of 1 442 VLBW/ELBW infants were collected from 24 NICUs in 2017. The median antibiotic use duration was 17 days (range: 0-86 days), accounting for 53.0% of the total length of hospital stay. The highest duration of antibiotic use was up to 91.4% of the total length of hospital stay, with the lowest at 14.6%. In 16 out of 24 NICUs, the antibiotic use duration was accounted for more than 50.0% of the hospitalization days. There were 113 cases with positive bacterial culture grown in blood or cerebrospinal fluid, making the positive rate of overall bacterial culture as 7.84%. The positive rate of bacterial culture in different NICUs was significantly different from 0% to 14.9%. The common isolated bacterial pathogens Klebsiella pneumoniae was 29 cases (25.7%); Escherichia coli 12 cases (10.6%); Staphylococcus aureus 3 cases (2.7%). The most commonly used antibiotics were third-generation of cephalosporins, accounting for 41.00% of the total antibiotics, followed by penicillins, accounting for 32.10%, and followed by carbapenems, accounting for 13.15%. The proportion of antibiotic use time was negatively correlated with birth weight Z-score and the change in weight Z-score between birth and hospital discharge (r=-0.095, -0.151 respectively, P<0.01), positively correlated with death/withdrawal of care (r=0.196, P<0.01).
		                        		
		                        			CONCLUSIONS
		                        			Antibiotics used for VLBW/ELBW infants in NICUs of Hunan Province are obviously prolonged in many NICUs. The proportion of routine use of third-generation of cephalosporins and carbapenems antibiotics is high among the NICUs.
		                        		
		                        		
		                        		
		                        			Anti-Bacterial Agents
		                        			;
		                        		
		                        			Birth Weight
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Infant
		                        			;
		                        		
		                        			Infant, Extremely Low Birth Weight
		                        			;
		                        		
		                        			Infant, Newborn
		                        			;
		                        		
		                        			Intensive Care Units, Neonatal
		                        			;
		                        		
		                        			Surveys and Questionnaires
		                        			
		                        		
		                        	
10.Clinical features of coronavirus disease 2019 in children aged <18 years in Jiangxi, China: an analysis of 23 cases.
Hua-Ping WU ; Bing-Fei LI ; Xiao CHEN ; Hua-Zhu HU ; Shu-Ai JIANG ; Hao CHENG ; Xin-He HU ; Jian-Xin TANG ; Fu-Chu ZHONG ; Ling-Wen ZENG ; Wei YU ; Yan YUAN ; Xian-Fei WU ; Yu-Ping LI ; Zong-Li ZHENG ; Tian-Bo PAN ; Zhi-Xing WU ; Jin-Feng YUAN ; Qiang CHEN
Chinese Journal of Contemporary Pediatrics 2020;22(5):419-424
		                        		
		                        			OBJECTIVE:
		                        			To study the clinical features of coronavirus disease 2019 (COVID-19) in children aged <18 years.
		                        		
		                        			METHODS:
		                        			A retrospective analysis was performed from the medical data of 23 children, aged from 3 months to 17 years and 8 months, who were diagnosed with COVID-19 in Jiangxi, China from January 21 to February 29, 2020.
		                        		
		                        			RESULTS:
		                        			Of the 23 children with COVID-19, 17 had family aggregation. Three children (13%) had asymptomatic infection, 6 (26%) had mild type, and 14 (61%) had common type. Among these 23 children, 16 (70%) had fever, 11 (48%) had cough, 8 (35%) had fever and cough, and 8 (35%) had wet rales in the lungs. The period from disease onset or the first nucleic acid-positive detection of SARS-CoV-2 to the virus nucleic acid negative conversion was 6-24 days (median 12 days). Of the 23 children, 3 had a reduction in total leukocyte count, 2 had a reduction in lymphocytes, 2 had an increase in C-reactive protein, and 2 had an increase in D-dimer. Abnormal pulmonary CT findings were observed in 12 children, among whom 9 had patchy ground-glass opacities in both lungs. All 23 children received antiviral therapy and were recovered.
		                        		
		                        			CONCLUSIONS
		                        			COVID-19 in children aged <18 years often occurs with family aggregation, with no specific clinical manifestation and laboratory examination results. Most of these children have mild symptoms and a good prognosis. Epidemiological history is of particular importance in the diagnosis of COVID-19 in children aged <18 years.
		                        		
		                        		
		                        		
		                        			Adolescent
		                        			;
		                        		
		                        			Betacoronavirus
		                        			;
		                        		
		                        			Child
		                        			;
		                        		
		                        			Child, Preschool
		                        			;
		                        		
		                        			China
		                        			;
		                        		
		                        			Coronavirus Infections
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Infant
		                        			;
		                        		
		                        			Pandemics
		                        			;
		                        		
		                        			Pneumonia, Viral
		                        			;
		                        		
		                        			Retrospective Studies
		                        			
		                        		
		                        	
            
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